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STUDIES ON THE CHARACTERISTICS OF PHYTOMASS-BASED VERMICOMPOSTS AND THEIR INFLUENCE ON THE PLANT GROWTH
Thesis submitted
for the award of the degree of
DOCTOR OF PHILOSOPHY
in
Environmental Technology
by
M. Karthikeyan, MSc., MPhil
Under the supervision of
Dr. S. Gajalakshmi, MSc., MPhil., Ph.D
Assistant Professor
And Co-guidance of
Prof. S. A. Abbasi, PhD, DSc, FNASc, FIIChE, FIE, PE
Senior Professor and Head
Centre for Pollution Control and Environmental Engineering Pondicherry (Central) University Puducherry - 605014, India
CERTIFICATE
This is to certify that Mr. M. Karthikeyan has carried out the work embodied in his thesis
entitled ‘Studies on the characteristics of phytomass-based vermicomposts and their
influence on the plant growth’ being submitted to Pondicherry University for the award of
the degree of Doctor of Philosophy in Environmental Technology. He has complied with all
the relevant academic and administrative regulations, and the thesis embodies a bonafide
record of the work done by him under our guidance. The work is original and has not been
submitted for the award of any certificate, diploma, or degree, of this or any other university.
Dr. S. Gajalakshmi Prof. S. A. Abbasi
Supervisor Co-guide
DECLARATION
I hereby declare that the thesis entitled “Studies on the characteristics of phytomass-based
vermicomposts and their influence on the plant growth” submitted to Pondicherry
University for the award of the degree of Doctor of Philosophy is a record of original work
done by me under the guidance of Dr. S. Gajalakshmi, Assistant Professor, and co-guidance
of Prof. S. A. Abbasi, Senior Professor and Head, Centre for Pollution Control and
Environmental Engineering, Pondicherry University, and that it has not formed the basis for
the award of any other degree, diploma, certificate or any other title by any university or
institution before.
Date:
Place: Puducherry (M. Karthikeyan)
It may be doubted whether there are many other animals which have played so important a part in the history of the world, as have these lowly organized creatures.
Charles. R. Darwin (1881)
To My Parents
To My Wife Vimala and
Daughter Aaradhana
Acknowledgements
“Kindness shown by those who weigh not what the return may be:
When you ponder right its merit, 'Tis vaster than the sea” - Thirukural
After years and years of receiving support and help from family, teachers and friends, finally
time has come to thank these wonderful people for their unselfish contribution.
First of all I thank my thesis supervisor Dr. S. Gajalakshmi for her care and encouragement
especially during the difficult times, and for having confidence in me and my work. “Mam,
without your support, I would have not achieved this stage. I have no words to express my
gratefulness, so I salute you, mam”.
I am grateful to Prof. S.A. Abbasi. His wide knowledge, „decades and decades of experience‟
in this field and his logical way of approaching problems have been of great value for me. His
genuine and open-minded curiosity to learn new things has inspired me all over these years.
I would also like to acknowledge my doctoral committee member, Dr. D. Senthilnathan for
his stimulating decisions and valuable comments on my work. Other than my work, he wanted
me to submit thesis early. I never had seen such a kind-hearted teacher in my life, “Thank you
sir”.
I thank Dr. Tasneem Abbasi and Er. S.Sudalai for their timely help, and they never denied or
delayed any assistance whenever I have requested them. I also sincerely thank my RGNF
committee member Dr. E.V. Ramasamy for his valuable input in my research work.
I own my sincere gratitude to all my Teachers and Professors who have shaped me to what I
am. Especially my Maths teacher Mr. S. Jayaraj, post-graduate teacher Dr. P. Suresh and
Dr. Christabelle E.G. Fernandes, without them I would have not been able to reach this
stage. I have been so blessed to get these wonderful people in my life.
பயன்தூக்கார் செய்த உதவி நயன்தூக்கின்
நன்மை கடலின் சபரிது
I am indebted to Mr. Prabanandham for his help and care all along, “Anna, you presence
always made me laugh”. Special thanks to Mr. Premaravind for helping me in fixing both lab
and personal needs.
I am also grateful to Mr. Karunakaran, Mr. Balamurugan and Mr. Vetrivel, Department of
Earth Science, for helping me in thin-sectioning and ICP analysis. I must thank Er. Ramasamy
and Mrs. S. Elisa Fathima, Central Instrumentation Facility, Mr. Anbalagan, Sophisticated
Analytical Instrument Facility, IITM, Chennai, and Mr. Adarsh, Sophisticated Test and
Instrumentation Centre, Cochin University, Cochin for technical support.
Many thanks to mera dost Ajay Harit for his contribution in all my experimental work. Also I
thank my friends Sudakaran (Ecology), Babuji (Management studies), Vinoth (Physical
Education) and Gnanavel (Chemistry) for being there for my needs and I really had a wonderful
time with them during these „Ph.D years‟. Heartfelt thanks to my beloved friends Baskaran
(Marine Biology), Anandha Dhanaselvan (CPEE), Purushothaman (Philosophy) and
Srinivasan (Alagappa University) for their help and support.
I cannot forget the help of M.Phil scholar Hariharan, who have worked with me day and night
to set the plant growth studies. I thank my friend Nayeem Shah (alias Peter) for his fun full
company, and also I thank Antony Godson, Nasser Hussain and Mohammad Ashraf for the
same. Many people in CPEE have decorated my memories of „Ph.D years‟, for this I should
thank Dr. R. Sanjeevi, Dr. J. Anuradha, Dr. G. Ponni, Dr. Manoj Makhiija, Mrs. Gurjeet
Kaur, Mrs. P. Poornima, Mr. T. Ganesh, Mr. T.V. Ananthaaraj and so many… :-)
No words to either thank or praise my Parents and Lord Almighty who gave me every possible
support throughout and holding my hand each step of the way. I thank my wife for her
patience, love and support. I cordially thank my sister. ‘Indu! you are my strength!’ I thank
GOD for having blessed me with such a loving sister.
Last but not least, I would like to thank my lovely worms for its great job! “Buddies! I will not
disturb you anymore! Enjoy!”
M.Karthikeyan
i
Contents
List of Tables ……………………………………………………………………………………... vii-x
List of Figures …………………………………………………………………………………….. xi-xii
Chapter 1. Introduction 01
Chapter 2. Vermicompost characteristics and its effect on soil and plant growth – state-of-
the-art
Abstract………………………………………………………………………………………... 05
1. Introduction ………………………………………………………………………………... 06
2. Types of vermicast and its composition …………………………………………………… 06
2.1. Types of vermicast ………………………………………………………………… 06
2.1.1. Based on place of deposition: surface and subterranean castings ………... 06
2.1.2. Based on shape of castings ……………………………………………….. 06
2.1.3. Based on size of the castings ……………………………………………... 10
2.2. Physical properties of vermicast …………………………………………………... 11
2.2.1. Color and odor ……………………………………………………………. 11
2.2.2. Bulk density ………………………………………………………………. 11
2.2.3. Pore space ………………………………………………………………… 13
2.2.4. Water-related properties ………………………………………………….. 14
2.3. Chemical properties of vermicast …………………………………………………. 15
2.3.1. Organic carbon ……………………………………………………………. 15
2.3.2. Macronutrients (primary) ………………………………………………..... 20
2.3.3. Macronutrients (secondary) ………………………………………………. 23
2.3.4. Micronutrients …………………………………………………………….. 25
3. Effect of ageing on the properties of vermicast …………………………………………… 29
4. Effect of vermicast on plant growth and soil ………………………………………………. 31
5. Conclusion …………………………………………………………………………………. 32
References ……………………………………………………………………………………. 33
Chapter 3. Ingestion of sand and soil by phytophagous earthworm Eudrilus eugeniae: a
finding of relevance to earthworm ecology as well as vermitechnology
Abstract ……………………………………………………………………………………….. 53
1. Introduction ………………………………………………………………………………... 53
2. Materials and methods ……………………………………………………………………... 54
3. Results and discussion ……………………………………………………………………... 56
3.1. Vermicast output ………………………………………………………………….. 56
3.2. Mortality …………………………………………………………………………... 58
ii
3.3. Average zoomass change per animal ……………………………………………... 58
3.4. Surface area of vermicast …………………………………………………...…….. 58
3.5. Assimilation of sand and soil particles in castings ……………………………… 59
4. Conclusions ………………………………………………………………………………... 61
References ……………………………………………………………………………………. 61
Chapter 4. Feeding behaviour of phytophagous earthworm Eudrilus eugeniae in high-
substrate column vermireactors
Abstract ……………………………………………………………………………………….. 65
1. Introduction ………………………………………………………………………...……… 65
2. Materials and methods …………………………………………………………...………… 66
3. Results and discussion …………………………………………………………………… 67
4. Conclusions ………………………………………………………………………………... 69
References ……………………………………………………………………………………. 69
Chapter 5. Effect of sand and soil ingestion by phytophagous earthworm Eudrilus eugeniae
on the physical and chemical properties of vermicast
Abstract ……………………………………………………………………………………….. 71
1. Introduction ………………………………………………………………………………... 71
2. Materials and methods ……………………………………………………...………….…... 72
2.1. Experimental design ………………………………………………………………. 72
2.2. Vermireactors operation …………………………………………………………... 72
2.3. Analytical methods ……………………………………………...………………… 72
2.4. Assessment of soil/sand content in the vermicast ………………………………… 73
2.5. Data analysis ……………………………………………………...……………….. 73
3. Results and discussion ……………………………………………………………………... 73
3.1. Assimilation of sand/soil in the vermicast ………………………………………… 73
3.2. Vermicast output ………………………………………………………………….. 75
3.3. Growth and survival of earthworms ………………………………………………. 75
3.4. Physical and chemical properties of vermicast …………………………………… 76
4. Conclusions ………………………………………………………………..………………. 77
References ……………………………………………………………………………………. 78
Chapter 6. Effect of vermicast generated from an allelopathic weed lantana (Lantana
camara) on seed germination, plant growth, and yield of cluster bean (Cyamopsis
tetragonoloba)
Abstract ……………………………………………………………………………………….. 81
1. Introduction ……………………………………………………………………………… 81
2. Materials and methods ……………………………………………………………………... 82
2.1. Germination, plant growth and yield characteristics ……………………………… 83
2.3. Analytical methods ………………………………………...……………………… 84
iii
2.4. Statistical analysis …………………………………………………….………… 85
3. Results and discussion …………………………………………………………………… 85
3.1. Seed germination ………………………………………………………………… 85
3.2. Plant growth …………………………………………..…………………………… 86
3.3. Photosynthetic pigments ……………………………..…………………………… 88
3.4. Flowering ………………………………………………………………………….. 88
3.5. Disease incidence, plant death and stunted growth ……………………………….. 88
3.5. Fruit yield ………………………………………….……………………………. 89
3.6. The present study in the context of the state-of-the-art ………...…………………. 90
4. Conclusions ………………………………………………………………………………. 91
References ……………………………………………………………………………………. 91
Supplementary material ………………………………………………………………............. 96
Chapter 7. Effect of vermicast generated from a pernicious weed ipomoea (Ipomoea
carnea) on seed germination, plant growth, and yield of cluster bean (Cyamopsis
tetragonoloba)
Abstract ……………………………………………………………………………………….. 101
1. Introduction ……………………………………………...………………………………… 101
2. Materials and methods ……………………………………………………………………. 102
2.1. Germination, plant growth and yield characteristics ……………………………… 103
2.3. Analytical methods …………………………………...…………………………… 104
2.4. Statistical analysis ………………………………………………………………… 104
3. Results and discussion …………………………………………………………………….. 104
3.1. Seed germination ………..………………………………………………………… 104
3.2. Plant growth …………….…………………………………………………………. 105
3.3. Photosynthetic pigments …...……………………………………………………… 107
3.4. Effect on flowering …………...…………………………………………………… 107
3.5. Disease incidence, plant death and stunted growth ……………………………….. 108
3.6. Effect on yield …………………………………………………………………….. 108
4. Conclusions ………………………………………..………………………………………. 109
References ……………………………………………………………………………………. 109
Supplementary material ………………………………………………………………………. 112
Chapter 8. Comparative efficacy of vermicomposted paper waste and inorganic fertilizer
on seed germination, plant growth and fruition of cluster bean (Cyamopsis tetragonoloba)
Abstract ……………………………………………………………………………………….. 117
1. Introduction ………………………………………………………………………………... 117
2. Materials and methods …………………………………………………………………… 118
2.1. Study area …………………………………………………………………………. 118
2.2. Treatments ………………………………………………………………………… 118
2.3. Germination, plant growth and yield characteristics ……………………………… 119
iv
2.3. Analytical methods ………………………………………………………………... 120
2.4. Statistical analysis ………………………………………………………………… 120
3. Results and discussion ……………………………………………………………………... 120
3.1. Seed germination ………………………………………………………………... 120
3.2. Plant growth ……………………………………………………………………... 121
3.3. Photosynthetic pigments ………………………………………………………… 123
3.4. Disease incidence, plant death and stunted growth ……………………………….. 123
3.5. Effect on flowering ……………………………………………………………… 123
3.6. Effect on yield …………………………………………………………………….. 123
4. Conclusions ………………………………………………………………………………... 124
References ……………………………………………………………………………………. 124
Supplementary material ………………………………………………………………………. 127
Chapter 9. Effect of vermicast generated from allelopathic weeds and paper waste on
physical and chemical properties of potting soil growing cluster bean (Cyamopsis
tetragonoloba)
Abstract ………………………………………………………………………………………. 131
1. Introduction ……………………………………………………………………………….. 131
2. Materials and methods …………………………………………………………………….. 132
3. Results and discussion ………………………………………………………………….. 134
3.1. Physical properties ………………………………………………………………. 134
3.2. Chemical properties …………………………………………………………….. 142
4. Conclusions ………………………………………………………………………………. 154
References …………………………………………………………………………………… 155
Chapter 10. Effect of storage on some physical and chemical characteristics of vermicast:
A preliminary study
Abstract ……………………………………………………………………………………… 159
1. Introduction ……………………………………………………………………………… 159
2. Materials and methods ……………………………………………………………………... 160
3. Results and discussion ……………………………………………………………………... 161
4. Conclusions ………………………………………………………………………………... 164
References ……………………………………………………………………………………. 164
Chapter 11. Effect of storage on the properties of vermicompost generated from paper
waste – with focus on pre-drying and extent of sealing
Abstract ……………………………………………………………………………………….. 169
1. Introduction ………………………………………………………………………………... 169
2. Materials and methods ……………………………………………………………………... 170
2.1. Types of storage ………………………………………………………………… 170
2.2. Analysis …………………………………………………………………………… 171
v
2.3. Processing of data …………………………………………………………………. 171
3. Results and discussion …………………………………………………………………… 171
3.1. Physical properties ………………………………………………………………… 171
3.2. Chemical properties ……………………………………………………………….. 173
3.3. Biochemical properties ……………………………………………………………. 177
4. Conclusions ……………………………………………………………………………… 180
References …………………………………………………………………………………… 180
Chapter 12. Effect of pre-drying and extent of sealing on the properties of vermicast
generated from the neem leaves during storage
Abstract ……………………………………………………………………………………….. 185
1. Introduction ………………………………………………………………………………. 185
2. Materials and methods ……………………………………………………………………... 186
2.1. Experimental set up ……………………………………………………………….. 186
2.2. Analytical methods ………………………………………………………………... 186
2.3. Data analysis ………………………………………………………………………. 186
3. Results and discussion ……………………………………………………………………... 186
3.1. Physical properties ………………………………………………………………… 186
3.2. Chemical properties ……………………………………………………………….. 190
3.3. Biochemical properties ……………………………………………………………. 194
4. Conclusions ………………………………………………………………………………... 198
References ……………………………………………………………………………………. 198
Chapter 13. Effect of pre-drying and extent of sealing on the properties of vermicomposted
cow dung during storage
Abstract ……………………………………………………………………………………….. 203
1. Introduction ………………………………………………………………………………... 203
2. Materials and methods …………………………………………………………………… 203
2.1. Experimental design ………………………………………………………………. 203
2.2. Analytical methods ………………………………………………………………... 204
2.3. Data analysis ………………………………………………………………………. 204
3. Results and discussion ……………………………………………………………………... 204
3.1. Physical properties ………………………………………………………………… 204
3.2. Chemical properties ……………………………………………………………….. 206
3.3. Biochemical properties ……………………………………………………………. 212
4. Conclusions ………………………………………………………………………………... 214
References …………………………………………………………………………………..... 215
Chapter 14. Summary and conclusion 219
Appendix – Standardization of analytical methods 223
vi
vii
List of Tables Chapter 2
1. Shape and size of the castings produced by different earthworm species ……………………. 08
2. Some of the chemical properties of castings produced by different earthworm species from
different organic wastes. ……………………………………………………………………… 17
3. Some of the micronutrient content of castings produced by the different earthworm species
from the different organic wastes…………………………………………………………… 27
Chapter 3
1. Major and trace element composition of plant leaves, cow dung, soil and sand used in this
study …………………………………………………………………………………………. 55
2. Repeated analysis of variants and ANOVA table of F-values and the effects of substrate,
bedding and worm density on vermicast output, average zoomass changes, mortality, sand
and soil entrapped in castings, castings surface area and sand grains covered area of
castings………………………………………………………………………………………... 57
Chapter 5
1. The physical properties of vermicast generated from the reactors without sand/soil + 250
and 500 g substrate and reactors consisting sand/soil + 250 and 500 g substrate…...……...… 76
2. The chemical properties of vermicast generated from the reactors without sand/soil + 250
and 500 g substrate and reactors consisting sand/soil + 250 and 500 g substrate………..…… 77
Chapter 6
1. Chemical and physical properties of vermicast and soil used in the study ………………… 84
2. Amount of inorganic fertilizer applied equivalent to vermicast treatment……………………. 85
3. Germination value and germination percentage of the seeds of cluster bean as influenced by
lantana vermicast and inorganic fertilizers…………………………………………………… 86
4. Plant growth, leaf pigments, flowering, disease incidence, and retarded growth stunned
growth/death in cluster bean plants as impacted by lantana vermicast or inorganic
fertilizers……………………………………………………………………………………… 87
5. Harvest index and yield attributes of plants as impacted by lantana vermicast and equivalent
inorganic fertilizers……………………………………….…………………………………… 89
Chapter 7
1. Chemical and physical properties of vermicast and soil used in the study ………………… 103
2. Amount of inorganic fertilizer applied equivalent to vermicast treatment …………………… 104
3. Germination value and germination percentage of the seeds of cluster bean as influenced by
ipomoea vermicast and inorganic fertilizers………………………………….……………… 105
4. Plant growth, leaf pigments, flowering, disease incidence, and retarded growth stunned
growth/death in cluster bean plants as impacted by ipomoea vermicast or inorganic
fertilizers…………………………………………………………………………………….. 106
5. Harvest index and yield attributes of plants as impacted by ipomoea vermicast and
equivalent inorganic fertilizers……………………………………………………………….. 108
viii
Chapter 8
1. Chemical and physical properties of vermicast and soil used in the study …………………... 119
2. Amount of inorganic fertilizer applied equivalent to vermicast treatment …………………… 120
3. Germination value and germination percentage of the seeds of cluster bean as influenced by
paper waste vermicast and inorganic fertilizers……………….…………….……………… 121
4. Plant growth, leaf pigments, flowering, disease incidence, and retarded growth stunned
growth/death in cluster bean plants as impacted by paper waste vermicast or inorganic
fertilizers………………………...…………………………………………………………….. 122
Chapter 9
1. Chemical and physical properties of vermicast and soil used in the study ………………… 133
2. Changes in the bulk density of potting soil amended with vermicast from lantana, ipomoea
and paper waste, or equivalent inorganic fertilizers, at different periods of time ……………. 135
3. Changes in the particle density of potting soil amended with vermicast from lantana,
ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of time … 136
4. Changes in the total porosity of potting soil amended with vermicast from lantana, ipomoea
and paper waste, or equivalent inorganic fertilizers, at different periods of time…………….. 137
5. Changes in the water filled pore space of potting soil amended with vermicast from lantana,
ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of time… 138
6. Changes in the water holding capacity of potting soil amended with vermicast from lantana,
ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of time…... 139
7. Changes in the electrical conductivity of potting soil amended with vermicast from lantana,
ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of time…... 140
8. Changes in the pH of potting soil amended with vermicast from lantana, ipomoea and paper
waste, or equivalent inorganic fertilizers, at different periods of time ……………………….. 141
9. Changes in the organic carbon of potting soil amended with vermicast from lantana,
ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of time… 144
10. Changes in the total nitrogen of potting soil amended with vermicast from lantana, ipomoea
and paper waste, or equivalent inorganic fertilizers, at different periods of time ……………. 145
11. Changes in the ammonia of potting soil amended with vermicast from lantana, ipomoea and
paper waste, or equivalent inorganic fertilizers, at different periods of time ………………… 146
12. Changes in the nitrate of potting soil amended with vermicast from lantana, ipomoea and
paper waste, or equivalent inorganic fertilizers, at different periods of time ………………… 147
13. Changes in the exchangeable phosphorus of potting soil amended with vermicast from
lantana, ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of
time …………………………………………………………………………………………… 148
14. Changes in the exchangeable potassium of potting soil amended with vermicast from
lantana, ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of
time …………………………………………………………………………………………… 149
15. Changes in the exchangeable calcium of potting soil amended with vermicast from lantana,
ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of time….. 150
16. Changes in the trace elements of potting soil amended with vermicast from lantana or
equivalent inorganic fertilizers, at different periods of time………………………………… 151
17. Changes in the trace elements of potting soil amended with vermicast from ipomoea or
equivalent inorganic fertilizers, at different periods of time …………………………………. 152
18. Changes in the trace elements of potting soil amended with vermicast from paper waste or
equivalent inorganic fertilizers, at different periods of time …………………………………. 153
ix
Chapter 10
1. Physical characteristics of castings stored for different periods and the calculated F-values
using one-way ANOVA ……………………………………………………………………… 162
2. Total nitrogen and organic carbon of castings stored for different periods and the calculated
F-values using one-way ANOVA ……………………………………………………………. 162
3. Major elements present in the casting stored for different periods …………………………... 163
Chapter 11
1. F values of repeated measures analysis of variance on the effect of extend of sealing and
pre-treatment on physical properties, EC and pH of vermicast during the storage…………… 174
1. F values of repeated measures analysis of variance on the effect of extend of sealing and
pre-treatment on chemical properties of vermicast during the storage……………………….. 174
2. F values of repeated measures analysis of variance on the effect of extend of sealing and
pre-treatment on biochemical properties of vermicast during the storage……………………. 174
Chapter 12
1. F values of repeated measures analysis of variance on the effect of extend of sealing and
pre-treatment on physical properties, EC and pH of vermicast during the storage…………… 189
2. F values of repeated measures analysis of variance on the effect of extend of sealing and
pre-treatment on chemical properties of vermicast during the storage……………………….. 189
3. F values of repeated measures analysis of variance on effect of extend of sealing and pre-
treatment on biochemical properties of vermicast during the storage………………………… 189
Chapter 13
1. F values of repeated measures analysis of variance on the effect of extend of sealing and
pre-treatment on physical properties, EC and pH of vermicast during the storage…………… 209
2. F values of repeated measures analysis of variance on the effect of extend of sealing and
pre-treatment on chemical properties of vermicast during the storage……………………….. 209
3. F values of repeated measures analysis of variance on the effect of extend of sealing and
pre-treatment on biochemical properties of vermicast during the storage.…………………… 209
x
xi
List of Figures Chapter 3
1. Vermicast output, g worm-1
day-1
recorded in reactors with different treatments ……………. 56
2. Percentage of sand and soil particle entrapped in the castings measured by gravimetric
method and percentage of sand particle covered area in the castings measured by thin-
sectioning method …………………………………………………………………………….. 59
3. The thin sectioned vermicast from neem, cow dung and ipomoea based reactors with or
without soil + sand bedding……………….…..………………………………………………. 60
4. Regression analysis between the sand and soil entrapped in the castings and (a) the average
zoomass changes and (b) mortality rate …………………………………………………….... 60
Chapter 4
1. Percentage of sand and soil particle entrapped in the castings of different treatments ………. 67
2. Vermicast output, grams worm-1
day-1
recorded in reactors with different treatment ………... 68
Chapter 5
1. Percentage of sand and soil entrapped in the vermicast from reactors consisting sand/soil
with 250 g and 500 g substrate in the first trial ………………………………………………. 73
2. Percentage of sand and soil entrapped in the vermicast from reactors consisting sand/soil
with 250 g and 500 g substrate in the second trial …………………………………………... 74
3. Vermicast output, grams worm -1 day-1 recorded in reactors without sand/soil + 250 and
500 g substrate and reactors consisting sand/soil + 250 and 500 g substrate ………………… 75
Chapter 11
1. Changes in the moisture content (a), bulk density (b), particle density (c) and water holding
capacity (d) of un-dried and pre-dried castings stored in airtight sealed bags and un-dried
and pre-dried castings stored in partially sealed bags, at different periods of time ………….. 172
2. Changes in the total porosity (a), water-filled porosity (b), pH (c) and EC (d) of un-dried and
pre-dried castings stored in airtight sealed bags and un-dried and pre-dried castings stored in
partially sealed bags, at different periods of time …………………………………………….. 173
3. Changes in the total organic carbon (a), dissolved organic carbon (b), total nitrogen (c),
ammonium nitrogen (d), nitrate nitrogen (e) and available phosphorus (f) content of un-dried
and pre-dried castings stored in airtight sealed bags and un-dried and pre-dried castings
stored in partially sealed bags, at different periods of time …………………………………... 176
4. Changes in the extractable form of potassium (a), sulfur (b), calcium (c) and sodium (d)
content of un-dried and pre-dried castings stored in airtight sealed bags and un-dried and
pre-dried castings stored in partially sealed bags, at different periods of time ………………. 177
5. Changes in the dehydrogenase (a), β- glucosidase (b), cellulase (c), urease (d), alkaline
phosphatase (e) and arylsulphatase (f) enzymes activity of un-dried and pre-dried castings
stored in airtight sealed bags and un-dried and pre-dried castings stored in partially sealed
bags, at different periods of time …………………………………………………………… 178
6. Changes in the microbial biomass carbon content of un-dried and pre-dried castings stored
in airtight sealed bags and un-dried and pre-dried castings stored in partially sealed bags, at
different periods of time ……………………………………………………………………… 179
xii
Chapter 12
1. Changes in the moisture content (a), bulk density (b), particle density (c) and water holding
capacity (d), of un-dried and pre-dried castings stored in airtight sealed bags and un-dried
and pre-dried castings stored in partially sealed bags, at different periods of time ………… 187
2. Changes in the total porosity (a), water-filled porosity (b), pH (c) and EC (d), of un-dried
and pre-dried castings stored in airtight sealed bags and un-dried and pre-dried castings
stored in partially sealed bags, at different periods of time ………………………………… 188
3. Changes in the total organic carbon (a), dissolved organic carbon (b), total nitrogen (c),
ammonium nitrogen (d), nitrate nitrogen (e) and available phosphorus (f), of un-dried and
pre-dried castings stored in airtight sealed bags and un-dried and pre-dried castings stored in
partially sealed bags, at different periods of time …………………………………………. 191
4. Changes in the extractable form of potassium (a), sulfur (b), calcium (c) and sodium (d) of
un-dried and pre-dried castings stored in airtight sealed bags and un-dried and pre-dried
castings stored in partially sealed bags, at different periods of time ……………………..... 192
5. Changes in the dehydrogenase (a), β- glucosidase (b), cellulase (c), urease (d), alkaline
phosphatase (e) and arylsulphatase (f) enzymes activity of un-dried and pre-dried castings
stored in airtight sealed bags and un-dried and pre-dried castings stored in partially sealed
bags, at different periods of time ……………………………………………………………... 195
6. Changes in the microbial biomass carbon content of un-dried and pre-dried castings stored
in airtight sealed bags and un-dried and pre-dried castings stored in partially sealed bags, at
different periods of time ……………………………………………………………………… 197
Chapter 13
1. Changes in the moisture content (a), bulk density (b), particle density (c) and water holding
capacity (d), of un-dried and pre-dried castings stored in airtight sealed bags and un-dried
and pre-dried castings stored in partially sealed bags, at different periods of time ………… 205
2. Changes in the total porosity (a), water-filled porosity (b), pH (c) and EC (d), of un-dried
and pre-dried castings stored in airtight sealed bags and un-dried and pre-dried castings
stored in partially sealed bags, at different periods time …………………………………… 206
3. Changes in the total organic carbon (a), dissolved organic carbon (b), total nitrogen (c),
ammonium nitrogen (d), nitrate nitrogen (e) and available phosphorus (f), of un-dried and
pre-dried castings stored in airtight sealed bags and un-dried and pre-dried castings stored in
partially sealed bags, at different periods of time …………………………………………. 207
4. Changes in the extractable form of potassium (a), sulfur (b), calcium (c) and sodium (d) of
un-dried and pre-dried castings stored in airtight sealed bags and un-dried and pre-dried
castings stored in partially sealed bags, at different periods of time …………………………. 210
5. Changes in the dehydrogenase (a), β- glucosidase (b), cellulase (c), urease (d), alkaline
phosphatase (e) and arylsulphatase (f) enzymes activity of un-dried and pre-dried castings
stored in airtight sealed bags and un-dried and pre-dried castings stored in partially sealed
bags, at different periods of time …………………………………………………………… 211
6. Changes in the microbial biomass carbon content of un-dried and pre-dried castings stored
in airtight sealed bags and un-dried and pre-dried castings stored in partially sealed bags, at
different periods of time …………………………………………………………………….... 214
INTRODUCTION
Chapter
1
1
CChhaapptteerr 11
Introduction
Vermicomposting has become increasingly
popular across the world in recent decades
(Gajalakshmi and Abbasi, 2008; Edwards et al.,
2011). The ability of this process to convert various
types of biodegradable solid wastes into organic
fertilizer in a relatively low energy consuming and
pollution-generating manner has been the reason
behind this happening. Vermicompost is stabilized
organic waste by microbial bio-oxidation with
specific mediation of earthworms. It is reported to
improve seed germination, enhance seedling
growth and development, and increase plant
productivity even with a relatively small proportion
incorporated into growing media (Atiyeh et al.,
1998; Edwards et al., 2011). The extent of these
beneficial impacts of vermicompost on plants
depends on various factors. The nutrient content,
microbial activity, hydraulic properties and growth
regulators present in the vermicompost are crucially
important. All these beneficial properties of
vermicompost are subject to various changes with
substrate type, earthworm species and age of
castings. The dynamics of all these properties of
vermicast and its impact on plant growth and soil is
reviewed in Chapter 2.
The Chapters 3-5 comprise of the studies on
the feeding behavior of epigeic earthworms, which
are the species extensively used for
vermicomposting of various types of organic waste
(Abbasi and Ramasamy, 2001; Gajalakshmi et al.,
2001, 2002, 2005; Gajalakshmi and Abbasi,
2004a,b; Garg and Kaushik, 2005; Lim et al., 2012,
2014; Shak et al., 2014). The epigeics are
phytophagous earthworms which dwell at or very
near the surface of the soil horizon and feed upon
humus. The other two groups – anecics and
endogeics – make deep burrows, often live in
subsoil and due to their tendency to ingest soil they
are called as geophytophagous and geophagous
respectively. Many studies on anecics and
endogeics claim that ingestion of soil particles with
organic matter facilitates assimilation of nutrients
in earthworms gut probably by enhancing the
grinding action of the gizzard (Hendriksen, 1991;
Schulmann and Tiunov, 1999; Marhan and Scheu,
2005; Curry and Schmidt, 2007). But there are no
reports on the ingestion of sand particles by epigeic
earthworms. To fill this knowledge gap, attempt has
been taken to explore the influence of different
phytomass and worm density on ingestion of
soil/sand particles by epigeics and reported in
Chapters 3 and 4. The effect of mineral soil particle
ingestion on physico-chemical properties of
vermicast is studied and reported in Chapter 5.
In general, vermicast contains all the
essential macro and micronutrients for plant
growth, and it also improves the physical properties
of the soil, such as aeration, water holding capacity
and porosity of soil, all of which have direct impact
on the plant productivity (Edwards, 2004;
Gajalakshmi and Abbasi, 2008; Edward et al.,
2011). But it has to be ascertained whether the
vermicast generated from the invasive plants are
also beneficial to plants, as they are widely reported
to suppress the growth of other plant species.
Therefore, attempt has been made to assess the
effect of vermicompost generated from allelopathic
weeds Lantana camara and Ipomoea carnea on the
germination and growth of cluster bean (Cyamopsis
tetragonoloba) and reported in Chapters 6-8. The
response of this plant to vermicompost generated
from these weeds has been compared with
2
vermicompost from paper waste and equivalent
inorganic fertilizer treatments. The findings of these
studies revealed that the vermicompost derived
from different parent materials have different
physical, chemical and biological qualities, and
their impact on germination, growth and yield of
plant also varied considerably. To understand the
factors which attribute the differential impact of
vermicompost from different parent materials on
the growth and yields of plants, it is necessary to
recognize their impact on soil health. Hence, the
changes in the physical and chemical properties of
potting soil housing cluster bean (Cyamopsis
tetragonoloba) with the application of vermicast
generated from different organic wastes such as
paper waste, leaves of ipomoea (Ipomoea carnea),
and of lantana (Lantana camara) was studied and
reported in Chapter 9.
Although there are numerous reports on
vermicomposting and its application, there seems to
be an urgent need for the study of storage aspects of
vermicompost. There is a necessity to assess what
changes occurs in physico-chemical and biological
properties of vermicast during storage, so as to
work out the best strategy for packing and storing
the vermicompost which can enable retention of its
fertilizer value for long duration. In this regard,
attempts have been made towards formulating
packing guidelines for storing vermicast. The
findings of the study are briefed in the Chapters 10-
13. The summary of the findings of all the studies is
presented in Chapter 14. A brief account on the
standardization of analytical methods used in the
experiments reported in Chapters 3-13 is given in
Chapter 15.
References
Abbasi, S.A., Ramasamy, E.V., 2001. Solid waste
management with earthworms. Discovery
Publishing House, New Delhi. p.178.
Atiyeh, R.M., Subler, S., Edwards, C.A., 1998.
Growth of tomato plants in vermicomposted
hog manure. In proceedings of the 6th
International Symposium on Earthworm
Ecology Pedobiologia 43, 724–728.
Curry, J.P., Schmidt, O., 2007. The feeding ecology
of earthworms - a review. Pedobiologia 50,
463-477.
Edwards, C.A., Arancon, N.Q., Sherman, R., 2011.
Vermiculture technology: earthworms, organic
wastes and environmental management. CRC
Press, Boca Raton. pp. 103–164.
Gajalakshmi, S., 2002. Development of methods
for treatment and reuse of municipal and
agricultural solid wastes appropriate for
rural/suburban households. Ph.D thesis,
Pondicherry University, Puducherry. pp. 187.
Gajalakshmi, S., Abbasi, S.A., 2004a. Vermi-
conversion of paper waste by earthworm born
and grown in the waste-fed reactors compared
to the pioneers raised to adulthood on cow
dung feed. Bioresour. Technol. 94: 53–56.
Gajalakshmi, S., Abbasi, S.A., 2004b. Neem leaves
as a source of fertilizer-cum-pesticide
vermicompost. Biores. Technol. 92, 291-296.
Gajalakshmi, S., Abbasi, S.A., 2008. Solid waste
management by composting: state of the art.
Crit. Rev. Environ. Sci. Technol. 38, 311-400.
Gajalakshmi, S., Ramasamy, E.V., Abbasi, S.A.,
2001. Assessment of sustainable vermi-
conversion of water hyacinth at different
reactor efficiencies employing Eudrilus
eugeniae Kinberg. Bioresour. Technol. 80,
131-135.
Gajalakshmi, S., Ramasamy, E.V., Abbasi, S.A.,
2002. Vermicomposting of different forms of
water hyacinth by the earthworm Eudrilus
eugeniae, Kinberg. Bioresour. Technol.
82:165–169.
Gajalakshmi, S., Ramasamy, E.V., Abbasi, S.A.,
2005. Composting–vermicomposting of leaf
litter ensuing from the trees of mango
(Mangifera indica). Bioresour. Technol.
96:1057–1061.
Ganesh, P.S., 2007. Some application of bioprocess
engineering in solid waste management, Ph.D
thesis, Pondicherry University, Puducherry.
pp. 182.
3
Hendriksen, N.B., 1990. Leaf litter selection by
detritivore and geophagous earthworms. Biol.
Fert. Soils 10, 17-21.
Kumar, T.G., 2009. Vermicomposting of pernicious
weed salvinia (Salvinia molesta, Mitchell).
M.Phil thesis, Pondicherry University,
Puducherry. pp. 88.
Makhija, M., 2012. Vermicomposting of solid
waste: Leaf litter, ipomoea, and used paper.
Ph.D thesis, Pondicherry University,
Puducherry. pp. 106.
Marhan, S., Scheu, S., 2005. Effects of sand and
litter availability on organic matter
decomposition in soil and in casts of
Lumbricus terrestris L. Geoderma 128, 155-
166.
Schulmann, O.P., Tiunov, A.V., 1999. Leaf litter
fragmentation by the earthworm Lumbricus
terrestris L. Pedobiologia 439, 453-458.
Garg, V.K., Kaushik, P., 2005. Vermistabilization
of textile mill sludge spiked with poultry
droppings by an epigeic earthworm Eisenia
foetida. Bioresour. Technol. 96, 1063-1071.
Lim, S.L., Wu, T.Y., Sim, E.Y.S., Lim, P.N.,
Clarke, C., 2012. Biotransformation of rice
husk into organic fertilizer through
vermicomposting. Ecol. Eng. 41, 60-64.
Lim, S.L., Wu, T.Y., Clarke, C., 2014. Treatment
and biotransformation of highly polluted agro-
industrial wastewater from a palm oil mill into
vermicompost using earthworms. J. Agric.
Food Chem. 62, 691-698.
Shak, K.P.Y., Wu, T.Y., Lim, S.L., Lee, C.A.,
2014. Sustainable reuse of rice residues as
feedstocks in vermicomposting for organic
fertilizer production. Environ. Sci. Pollut. Res.
21, 1349-1359.
4
VERMICOMPOST CHARACTERISTICS AND ITS EFFECT
ON SOIL AND PLANT GROWTH – STATE-OF-THE-ART
Chapter
2
5
CChhaapptteerr 22
Vermicompost characteristics and its effect on soil and plant
growth – state-of-the-art
Abstract
The beneficial effects of vermicast on soil productivity and plant growth has been appreciated since the early
work of Hensen (1877) and Darwin (1881). In general, the castings have low carbon and nitrogen ratio, high
porosity, and good water-holding capacity than soil. Vermicast contains most nutrients in forms that are
readily taken up by the plants. It is also reported to consist of plant growth-promoting substances. The
potential of vermicast to improve the physical, chemical and biological properties of soil and the subsequent
effect of these changes on plant growth have also been reported in both natural and agroecosystems. All
these beneficial properties of vermicast are subject to various changes with substrate type, earthworm species
and age of castings. In this chapter, dynamics of all these properties of vermicast and its impact on plant
growth and soil is reviewed.
1. Introduction
The structure of the soils and the
biogeochemical cycles associated with the soils are
strongly influenced by soil invertebrates (Lavelle et
al., 2006; Barrios, 2007). Among the soil
organisms, earthworms are unarguably the most
important in maintaining the soil integrity by
several means in various vegetated lands (Edwards,
2004). Though they are not dominant numerically,
they represent a major invertebrate biomass in soils
due to their much larger size in comparison to the
two other most influential soil-working organisms –
the ants and the termites. The physical and
chemical characteristics of the casts produced by
earthworms plays an important role in the
regulation of soil processes. The physical structure
and chemical linkage of soil and feed are
reorganized during the gut transit that leads to
modifications in their porosity, density, and
hydraulic-properties (Shipitalo and Protz, 1989;
Jouquet et al., 2008a). The contents of the feed are
also modified by gut microbes and enzymes that
increase mineralization of organic matter, and
therefore increase the plant available form of
nutrients in castings (Chapuis-Lardy et al., 2010;
Bityutskii et al., 2012).
Although the role of vermicast in the
regulation of soil processes and plant growth has
been explored in detail, little is known on how the
properties of castings vary with earthworm‘s
species and its life-traits and the feed they ingest
(Schrader and Zhang, 1997; Buck et al., 1999; Jana
et al., 2010). Moreover, the life-time and the
disintegration of these structures, which are of great
importance in nutrients dynamics, are yet to be
explored in detail.
This chapter reviews the (i) potential factors
that determine the properties of vermicast, (ii)
change in the biological, chemical, and physical
attributes of the vermicast with time, and (iii) the
impact of the factors stated above on plant growth
and soil.
6
2. Types of vermicast and its composition
2.1. Types of vermicast
Earthworms mainly ingest feed rich in
organic matter together with living
microorganisms, nematodes and other microfauna,
mesofauna and their dead remains. Most species
also consume mineral soil fractions at various
degrees and indicate that they prefer organic–
mineral mixtures than pure organic materials
(Doube et al., 1997). Earthworms digest these
materials with the help of the gut associated
microorganisms and assimilate nutrients from them.
The vermicast excreted after gut passage are of
various forms and exist with different features. The
nature of the castings is determined by earthworm
species, their habitat and feeding behavior.
Generally, the earthworms produce following type
of biogenic-structures.
2.1.1. Based on place of deposition: surface and
subterranean castings
Casts may be deposited on the surface of the
soil or within it, depending on the species (Lavelle,
1988), food source, and bulk density of soil they
inhabit (Binet and Le Bayon, 1999). Based on the
place of deposition, the castings can be classified
into two groups – (i) surface castings and (ii)
subterranean castings. All the earthworms‘ species,
epigeic, anecic as well as endogeic deposit castings
at soil‘s surface layer, which has significant impact
on the physical and chemical characteristics of soil.
These surface casts increase the roughness of
surface soil and in turn affects water runoff and
infiltration into soil (Binet and Le Bayon, 1999; Le
Bayon and Binet, 2001). However, the fresh
castings are of low stability, easily dispersible in
runoff water, and lead to soil erosion and loss of
sediment-associated nutrients (Sharpley and Syers,
1976; Sharpley et al., 1979; Shipitalo and Protz,
1988; Binet and Tréhen, 1992; Ganeshamurthy et
al., 1998; Buck et al., 1999; Le Bayon and Binet,
1999). Moreover, surface casts dry out quickly,
harden, and, if compact, are likely to limit root
penetration, thereby reducing the ability of plant
roots to obtain the nutrients stored inside the casts
until they are broken down.
Anecic and particularly endogeic earthworms
produce subterranean castings, which are deposited
on the burrow walls, within the burrow or into other
sorts of soil macropores (Brown et al., 2000). The
subterranean castings do not influence the surface
soil properties such as nutrient transport, infiltration
and soil erosion (Jouquet et al., 2008a). Unlike
surface castings, the subterranean castings remain
fresh and retain moisture for longer duration and
are protected from the main agents of physical
degradation such as raindrops, animal trampling
etc. Hence, the degradation of these belowground
casts is probably even slower than for surface casts.
For instance, the half-lives ranged from 2 to 11
months for castings of larger anecic species,
Martiodrilus sp. (Binet and Le Bayon, 1999;
Decaens, 2000; Mariani et al., 2007). This species
produced both surface and subterranean castings
with an average dry weight of 25 g (Decaens,
2000). The casts of this species are likely deposited
more prior to their abandonment from borrows,
which is commonly the case after inundation by
heavy rainfall (Mariani et al., 2007). The surface
castings of this species were reported to be stable
for up to 4 months, and to then sharply fall after 6
months because of physical degradation of the casts
(Decaens, 2000). The castings of compact type,
inhibit the root extension and other hand, if they are
of the decompact type allow roots to penetrate more
easily and uptake nutrients available to plants
(Edwards, 2004).
2.1.2. Based on shape of castings
The shape of the castings produced by
different species is dissimilar; their properties and
the impact on soil may therefore differ considerably
(Lavelle, 1988). Castings can be classified into four
major categories based on their shape. They are: (i)
globular, (ii) paste likes slurry, (iii) tall vertical
7
columns or heaps of different shapes, and
(iv)granular (Edwards, 2004). Production of
castings of any of these types appears to be related
to soil texture and the size of earthworm. Specific
anatomic features of earthworms have also been
identified that allow the worm either to release
large units at regular intervals that will form the
globular cast, or a thin constant flux of cast material
that will form the poorly structured granular casts
(Lavelle and Spain, 2003). The shape and size of
the castings produced by some earthworm species
has been summarized in Table 1.
(i) Globular castings: Globular castings
consist of coalescent round or flattened units, which
are denser than the surrounding soil (Edwards,
2004; Blouin et al., 2007). This type of castings is
produced by anecic and endogeic species which are
larger species. It is stable for several months. The
greater structural stability of this castings is
probably due to the intense mixing and compaction
of gut content with mucopolysaccharides before it
released, which glue the soil particles together and
produce compact stable-structures (Lavelle, 1988).
Moreover, the castings are excreted with some
colorless waxy fluid from their nephridia (Agarwal
et al., 1958), calcium humate and calcite generated
during the organic matter decomposion and
calciferous glands, respectively which all perhaps
function as the cementing material to the castings
and harden them (Oyedelea et al., 2006). These
stable globular castings improve water infiltration
due to enhancement of the soil surface roughness
and modification in the circulation. Being water
stable aggregates, the vermicast is prevented from
detachment of easily transportable particles, which
in turn reduces erosion (Jouquet et al., 2008a).
An anecic earthworm species, Amynthas
khami in the Northern Vietnam (Bouché, 1977), is
reported to build biogenic structures up to 20 cm
length, which are formed from continuous
deposition of globular casts (Jouquet et al., 2008b).
These structures are connected with belowground
galleries and aboveground casts (Jouquet et al.,
2008a). Similarly, the large size globular castings
were reported in Andiodrilus pachoensis, a large
anecic earthworm. They deposit casts in the form of
piles on the soil surface. The piles can reach up to
12 cm height with a central canal called the
earthworm gallery (Thomas et al., 2008). Casts are
deposited on several occasions less than one week.
Hence the old and fresh portion of vermicast can be
differentiated. But once the formation of casts
diminishes, this heterogeneity disappears.
(ii) Slurry type of castings: Slurry type of
biogenic structures is single masses of soil mainly
produced by anecics and endogeics (Edwards,
2004). These casts are without a distinct shape,
paste-like and unstable, but stabilize with ageing
(Marinissen and Dexter, 1990). Martiodrilus sp. a
large size, dorsally dark-grey pigmented, surface-
casting species is reported to produce this type of
castings. The size of these casts is large. The casts
are of 3-6 cm diameter, height 2 to 10 cm and 25 g
average dry weight. The biogenic structures are
become tower like shaped by dry material at its
base and fresh pasty material at the top (Jiménez
and Decaëns, 2004).
(iii) Tall vertical columns or heaps of
variable shape castings: This type of castings is
usually deposited by anecic or endogeic groups.
These are formed by the sequential deposition on
globular casts. They have a hole in the middle when
they are tower form (Edwards, 2004). This type of
castings has been reported with Martiodrilus
carimaguensis. The castings are up to 15 cm height
with 5 cm diameter. They form tower like structure
which are deposited in the course of several days
(Jiménez et al., 1998; Decaëns et al., 1999).
Another species of earthworm Hyperiodrilus
africanus, which is a predominantly surface casting
species in large parts of moist savannahs of West
Africa, produced same type of castings. H.
africanus travels in the same channel several times
to deposit ingested soil at the soil surface, forming
characteristic turret shaped casts (Hauser and
Asawalam, 1998). Large amount of turret shaped
8
Table 1. Shape and size of the castings produced by different earthworm species.
Earthworm species Average size of the
worms
Average
weight of the
worms (mg)
Shape of the castings Range of the size of
castings References
Amynthas khami Up to 500 mm length - Globular 100- 200 mm height x 30
mm diameter
Jouquet et al., 2008a,b;
Podwojewski et al., 2008
Andiodrilus pachoensis length of 200 mm -
Globular and central
canal which corresponds to the
earthworm gallery
120 mm height Thomas et al., 2008
Dichogaster jaculatrix 10 or 25 mm length ×
1.4 mm diameter - Tower like structure
100 -120 mm height x 40
mm diameter Baylis, 1915
Drawida papillifer 60-120 mm length ×
3-5mm diameter 200-730
Composite irregular paste like
slurries
10-15 mm height × 8-10
mm diameter Chaudhuri et al., 2008,2009
Eudrilus eugeniae 90- 416.3 mm length ×
4-8 mm width 230- 5923
Fine granular pellets deposited
small heaps, that may be 20 to
30 mm high and 30-50 mm
diameter
30 mm height x 30-50
mm diameter
Madge, 1969; Viljoen and
Reinecke, 1994; Kale, 1998;
Parthasarathi, 2007; Mainoo et al.,
2008
Eutyphoeus assamensis 260-275mm length ×
5-6 mm diameter 1210-3730 Tower like structure
30-50 mm height × 15-30
mm diameter Chaudhuri et al., 2009
Eutyphoeus callosus 310-410 mm length ×
8-10 mm diameter 8200-15800
Tower like castings with
compact and thick convolutions
40-55 mm height × 60-65
mm diameter Chaudhuri et al., 2009
Eutyphoeus comillahnus 120-165 mm × 3-5 mm
diameter 1000-2100
Tower like casts of fragile
aggregates with or without
convolutions
40-50 mm height ×25-30
mm diameter Chaudhuri et al., 2009
Eutyphoeus gammiei 200-400 mm length ×
7-10 mm diameter 8350-13200
Tower like castings with
compact and thick convolutions
140-160 mm height × 40-
50 mm diameter
Bhattacharjee and Chaudhuri,
2002; Chaudhuri et al., 2008,
2009.
Eutyphoeus gigas 145-290 mm length ×
7-11 mm diameter 2230-5200
Tower like castings with
compact and thick convolutions
50-70 mm height × 40-50
mm diameter Chaudhuri et al., 2008, 2009
Eutyphoeus scutarius 244-332 mm length ×
6-7 mm diameter 4750-8180
Tower like castings with
compact and thick convolutions
35-70 mm height × 20-35
mm diameter Chaudhuri et al., 2009
Eutyphoeus sp 80-195 mm length ×
3-4 diameter 500-1280
Tower like casts of fragile
aggregates with or without
convolutions
8-10 mm height × 20-25
mm diameter Chaudhuri et al., 2009
Eutyphoeus turaensis 135-190 mm length ×
2.5-3 mm diameter 400-1160
Tower like castings with
compact and thin convolutions
30-35 mm height × 10-25
mm diameter Chaudhuri et al., 2009
9
Earthworm species Average size of the
worms
Average
weight of the
worms (mg)
Shape of the castings Range of the size of
castings References
Hippopera nigeriae - -
Pipe like composite cast, which
had an average bore diameter of
2.0 mm ranging from 0.9-3.6
mm
16-118 mm height x 1.5
mm diameter
Nye, 1955; Wilkinson and Aina,
1976.
Hyperiodrilus africanus 120-160 mm length -
Pipe like composite cast, with a
vertical hole running through it
but closed at the top.
25-90 mm height x 10-
30 mm diameter
Madge, 1969; Lee, 1985; Hauser
and Asawalam, 1998; Tondoh,
1998.
Kanchuria sp 175-280 mm length ×
2.5-6 mm diameter 1550-3450 Large globoid
20-45 mm height × 30-45
mm diameter Chaudhuri et al., 2008, 2009.
Martiodrilus
carimaguensis
and 194.3 mm in length
and 9.3 mm in diameter
11200 Tall vertical heaps (turricule) Up to 150 mm height x
up to 50 mm diameter
Jiménez et al., 1998; Decaëns,
2000; Tondoh and Lavelle, 2005;
Mariani et al., 2007a; Mainoo et
al., 2008.
Metapheretima
jocchana
500-670 mm length ×
9-10 mm diameter - Very large composite casts
50 mm height x 50 mm
diameter Lee, 1967; Tsai et al., 2004.
Metaphire houlleti 92-200 mm length ×
4-7 mm diameter 1000-3130
Tower-like casts with regular
arrangement of
spherical/subspherical
aggregates
35-70 mm height × 15-35
mm diameter Chaudhuri et al., 2009
Metaphire posthuma 120 mm in length ×
5 mm diameter -
Granular , which are mainly
found belowground 2 to 3 mm diameter Bottinelli et al., 2010
Microchaetus sp 1,800 mm length ×
16-18 mm diameter - Turret shaped
30-100 cm height and 1
m diameter
Edwards and Bohlen, 1996;
Henrot and Brussaard, 1997;
Blakemore et al., 2007.
Notoscolex birmanicus
760-1200 mm length ×
5 mm diameter
- Tower like structure 200 -240 mm height x up
to 40 mm diameter
Gates, 1961; Tembe and Dubash,
1961; Blakemore et al., 2007.
Perichata sp - - Tower like structure 90 mm height x 40 mm
diameter Darwin, 1881
Pontoscolex
corethrurus
50-150 mm length ×
4-6 mm diameter
350-800
Composite irregular paste like
slurries
10-15 mm height × 10-15
mm diameter
Moreno and Paoletti, 2003;
Dlamini and Haynes, 2004;
Chaudhuri et al., 2008, 2009.
10
casts deposition is also reported in Microchaetus
microchaetus (Henrot and Brussaard, 1997).
Usually many endogeics deposit casts for several
days in the same place as they exhibit restricted
movement and thus build spectacular tower-like
structures. Casts are piled up to 5-10 cm height in
the moist tropical forests of Africa. This is common
in many other tropical regions also (Lee, 1985).
Lavelle (1968) has reported similar, but less
spectacular piles at Lamto. These casts were
deposited by small Eudrilidae species continuously
for up to 30 hours at the same place (Lavelle,
1968).
(iv) Granular castings: Granular castings are
pellet like. They are produced mainly by smaller
earthworm species (epigeic, small endogeic, and
some anecic species) and distributed on or beneath
the soil surface (Edwards, 2004). The pellets are
small fine-textured, unlike globular castings and
hence are subjected to wash away by rain more
easily. Therefore, it favors sheet erosion (Lavelle,
1988). Examples for the granular casts depositing
earthworms are Eudrilidae sp. and Eudrilus
eugeniae, which produce castings of < 5 mm
diameter in size (Mulongoy and Bedoret, 1989;
Henrot and Brussaard, 1997; Hauser et al., 1998).
2.1.3. Based on size of the castings
Based on size, the castings can be categorized
into two types: (i) large size (>5mm) castings and
(ii) small size (<5 mm) castings. The large size
castings are egested by species of compacting
earthworm. They produce castings of higher bulk
density than the soil they inhabit. The high bulk
density is due to: (i) the formation of organo-
mineral bonds after mixing and chemical
transformations in the gut; (ii) the reabsorption of
water in the latter part of the earthworm intestine;
and (iii) the strong compaction by the tail muscles
when casts are expelled (McKenzie and Dexter,
1988). Casts of compacting species are stable in
nature as it creates anaerobic conditions inside the
castings which slows down the microbial
decomposition (Blanchart et al., 1993). The small
size castings are formed by decompacting
earthworms. They are of low bulk density. The
castings of decompacting earthworms are not stable
as that of the castings produced by compacting
species.
The larger size castings may be globular,
paste like slurries, tall vertical heaps or column
shaped castings, and small size castings is granular.
The production of both the types of casts by a
different earthworm group is favorable for
maintenance of the soil structure (Decaëns, 2000).
For example, in tropical agroecosystems, large
endogeics, such as Pontoscolex corethrurus or
Millsonia anomala, egests compact casts which are
large in size. These castings increase the proportion
of macroaggregates and bulk density of soil. This
cast might create anaerobic conditions inside the
castings and in turn slow down the decomposition
(Blanchart et al., 1993). Smaller decompacting
earthworm species (e.g. Eudrilidae) feed on these
large casts and consequently produce small and
unstable casts (0.5–5 mm) (Blanchart et al., 1999).
When decompacting species ingest casts of
compacting species, previously physically protected
carbon gets mineralized. On the other hand, where
there is ingestion of casts of decompacting species
by compacting species, the mineralizable carbon it
contains tends to be protected in their large casts
(Dickschen and Topp, 1987; Blanchart et al., 1999).
The activity of compacting earthworm species
alone lead to the formation of a compact surface
crust (Six et al., 2004).
The size of the castings has strong positive
correlation with the size of earthworm species that
produce them (Brussaard and Ferrera-Cerrato,
1997). The size of few earthworm species and the
size and shape of the castings that they produced is
listed in Table 1. Usually the size depend on the
species and range between 1-10 mm in diameter
(Oades, 1993). The sizes of the castings also may
be varying within same species, which rely on the
feed quality (Jiménez et al., 1998). For instance,
11
Martiodrilus sp, a large anecic species in the
Eastern Plains of Colombia, exhibit high variability
in their diet (Mariani et al., 2007). They produce
casts with low organic content but larger in size
(Jiménez et al., 1998). This is due to the availability
of low nutrient containing substrate as feed to
earthworm. As a result they ingest more quantity of
feed to fulfill their energetic requirements (Curry
and Schmidt, 2007). The large size of casts may
also influence oxygen-diffusion into them; the
mineralization of nitrogen, nitrification and oxygen
uptakes are slower in larger casts than the smaller
aggregates (Adu and Oades, 1978). Studies of
Sextone et al. (1985) also support this finding, in
which the oxygen content of a wet aggregate (18
mm diameter) near the aggregate center is 0% and
at the edge of the aggregate 21%.
2.2. Physical properties of vermicast
2.2.1. Color and odor
The color of the castings is a main factor to
distinguish the castings from the soil and other
components in field and vermicomposting process
(Lavelly, 1968; Garcìa and Fragoso, 2002). The
casts are completely dissimilar from the feed being
generally dark brown to black and mull humus-like
in appearance (Hervas et al., 1989; Brown et al.,
2000; Barros et al., 2001). Dramatic changes in
color of these castings are reported during aging.
Binet and Le Bayon (1999) reported three different
color castings in temperate cultivated soils. They
are brown, light brown and whitish. Those castings
may be produced from six different earthworms‘
species, reported in their study area. The brown
type is fresh casts, brown light are moderate aged
dry castings and whitish castings are oldest eroded
type. Depending on the climatic conditions the
casts may be covered by moss of white fungi giving
whitish appearance to the aged and disintegrated
castings (Binet and Le Bayon, 1999). Castings from
different horizons also show variation in their color.
The castings of Pheretima darnliensis derived from
the three different horizons showed three different
colors. The castings of P. darnliensis was dark
brown and grey-yellow when it was derived from O
and A horizon, respectively and castings of
intermediate color was also produced which
resembled a mix of the two soil layers (Gould et al.,
1987).
The vermicast has been reported to offer faint
pleasant earthy odor (Senesi, 1989; Szczech, 1999;
Dickerson, 2001; Sharma et al., 2005; Turnell et
al., 2007), may be due to the inhibition of the
growth of foul-smell forming microbes in the
decaying waste biomass when passed through
earthworm gut (Pierre et al., 1982). The anti-
microbial property of secretion of pharyngeal
glands, intestine, gizzard and crop, may also be the
reason for modulation of the diversity of the gut
microflora (Edwards and Fletcher, 1988). The
antibacterial factors of earthworms are also
reported in the coelomic fluid of Eisenia fetida
(Lassegues et al., 1989; Valembois et al., 1991) and
Lumbricus terrestris (Tuckova et al., 1986;
Anderson, 1988). Earthworms also create aerobic
conditions in the waste materials, prohibiting the
function of anaerobes which produce mercaptans
and hydrogen sulfide which gives foul odour
(Pedersen and Hendriksen, 1993; Sinha et al.,
2002).
2.2.2. Bulk density
The bulk density of castings also play an
important role regardless of nutrients they contain,
influencing the density and hydrological properties
of soil they exist. In general, soil with low bulk
density (0.7–1.8 g cm-3
) is desirable for plant
growth (Lal and Shukla, 2004), whether those are
agricultural crops, trees, or turf grass. Moreover,
soils of low bulk density have a greater water
infiltration rate which reduces storm water flow and
runoff. On the other hand, a high bulk density can
impede root penetration and reduce the air and
water circulation in the soil. Since the amount of
cast production is high on surface, its effect on bulk
density of surface layer is highly significant. The
12
amount of castings deposited by earthworms over
the soil surface may range from 2 to 50 tonnes per
year, and it may exceed 1000 tonnes per year when
the belowground castings are included, which rely
on the type of soil and land use (Lee and Foster,
1991). Ponomareva (1950) and van de Westeringh
(1972) have referred that earthworm casts
constitute up to 50% of soil aggregates in surface
layer in temperate pastures. The top layer of mull-
type forest (Kubiena, 1953) wooded savanna in
Ivory Coast (Lavelle, 1978), consisted of almost
entirely earthworm casts; whereas, James (1991)
estimation shows that surface castings produced in
Kansas tallgrass prairie is equivalent to 4-6% of
surface at the top 15 cm soil each year.
The bulk density of the castings greatly vary
with different earthworm species. Usually the bulk
density of castings produced by compacting species
is higher than of the soil they inhabit. Introduction
of a compacting species, Millsonia anomala in a
de-faunated savanna in Ivory Coast by temporary
flooding, increased the soil bulk density much
higher than of its initial within 20 months period
(Lavelle and Spain, 2003). As mentioned in section
2.1.3, the factors that increase the bulk density: (i)
the mixing and chemical transformations in the gut
which facilitates the bonding of organic matter and
mineral constituents; (ii) the reabsorption of water
in the latter part of the earthworm intestine; and (iii)
the strong compaction by the anus when casts are
ejected (McKenzie and Dexter, 1988). The smaller
de-compacting species produce smaller and fragile
castings, with lower bulk density due to the less
dense packing when castings are expelled (Six et
al., 2004). The micromorphological observation on
casts from the de-compacting species also showed
soil particles less densely packed and with a high
fraction of bacteria than those of compacting type
(Josehko et al., 1989). The bulk density of fresh
castings of de-compacting species, Aporrectodea
rosea was about 1.15 mg cm -3
, when it bred in a
sandy loam soil (McKenzie and Dexter, 1987). It is
much lower than the bulk density of Millsonia
anomala, a compacting species in Lamto‘s
savannas of Ivory Coast, which produced castings
with high bulk density of 1.8 mg cm−3
, when the
bulk density of surface soil was about 1.45 mg
cm−3
. A high sand content (75%), with the low clay
(7.5%) and organic carbon (1%) also might be
reason for the higher bulk density of castings of M.
anomala (Blanchart et al., 1993).
The bulk density of the castings may vary
with different soil type. For instance, earthworm
casts of temperate ecosystems have bulk density
lower than the surrounding soil (Larink et al., 2001;
Marashi and Scullion, 2003) while those from
tropical soils are denser than the former (Blanchart
et al., 1993, 1999; Decaëns et al., 2001; Oyedele et
al., 2006). Bulk density of an anecic, Martiodrilus
carimaguensis, in a native savanna was 10% higher
than the castings of the same species in an intensive
pasture. The higher proportion of organic matter in
the casts deposited in the pasture may be the reason
for the lower bulk density when compared to casts
of the savanna (Decaëns, 2000). Similarly, the bulk
density of castings produced by Lumbricus
terrestris was 1.33 and 1.54 mg cm-3
in sandy silt
and loamy silt soil, respectively (Josehko et al.,
1989). The difference in the bulk density of these
castings was probably due to their varying organic
carbon content. The organic residues have been
shown to buffer the effects of compaction as they
possess the ability to be compressed and then
springs back to its normal shape once the pressure
has released. This property of organic matter
prevents the compaction of clay fraction of soil
which is attached with organic matter. Therefore,
high organic matter levels make the soil less
susceptible to soil compaction (Wortman and Jasa,
2003). As the castings age, the bulk density either
decrease or increase, which depends on the species
which produced them.
13
2.2.3. Pore space
Porosity is a major constraint on other
physical, chemical and biological properties; it is of
primary importance in the regulation of tensile
strength, hydrological properties, gas diffusion,
microbial colonization, nutrient mineralization etc.
The earthworms alter the soil pore space through
their borrowing and casting activities (Edwards,
2004). The burrowing activity of earthworms
substantially alters soil macro pores (>1 mm),
which act as flow paths in soil (Bouche´ and Al-
Addan, 1997; Trojan and Linden, 1998). The
castings deposited affect mainly the meso- and
microporosity in soil. During the gut transit, the
pore space between organic and mineral particles
may get altered and creates packing void in the
deposited castings (Shipitalo and Protz, 1989;
Blanchart et al., 1993; Chauvel et al., 1999). Casts
has rather low amount of tubular, crack and almost
no packing void poroids as compared to the soil
(Blanchart et al., 2004; Jouquet et al., 2008b). The
structural rearrangement of the soil particles in the
intestine of earthworm would result in more fine
pores and fewer macropores in the castings
(Blanchart et al., 1993). These pores can be divided
into three size classes: (i) transmission (>50 µm),
(ii) storage (0.5 to 50 µm) and (iii) residual pores
(<0.5 µm). The storage and residual pores are
mainly found in the casts while the transmission
pores are located between the casts (Decaens et al.,
1999b). These transmission pores control the
transport of water and solutes and (Lamande et al.,
2003) the storage pores retain water with greater
tenacity particularly in the pores in the cortex of the
casts which are of 1-10 µm size (McKenzie and
Dexter, 1987).
All these three classes of pore space vary
considerably based on the earthworm species that
produce them, particularly the structure of
earthworm‘s anterior and posterior musculature
(Lapied and Rossi, 2000). In general, the casting of
de-compacting species contains more meso-pores
than those of compacting species. Even the pore
space in the castings of similar type may vary
significantly. For instance, the castings of a
compacting species, Millsonia anomala contain
predominantly mesopores ranging between 10 to 20
µm, whereas, the castings of a similar compacting
species, Pontoscolex corethrurus were all smaller
than 1 µm (Chauvel et al., 1997). Due to the results
of fewer meso-pores, the compacting species
reduced infiltration rates (Blanchart et al., 2004).
The pore space in the castings may greatly
influence the structure, and function of the
microorganisms and microfaunal communities and
interaction between them. Also the pores of
different size support different microbial
populations (Hattori, 1988). For example, pore
diameter larger than 90 µm favor the growth of
microarthropods, whereas, it is not supported in
smaller than 1.2 µm diameter pores (Vreeken-Bujis
et al., 1998). Pore sizes in the order of 1 µm
support the growth of bacterial population, and the
pore diameter of 30-90 µm enhance the nematode
population density (Hassink et al., 1993). The
greater percentage of pore size of casts of compact
earthworm species Millsonia anomala (Blanchart et
al., 1993) were less than 10 µm, which impedes
nematode colonization and predation (Elliott et al.,
1980). Visser (1985) and Anderson et al. (1984)
suggested that fungal growth is greatest in soils
with large pore sizes that can allow fungal
sporulation (Vreeken-Bujis et al., 1998). The
growth of protozoan population may also be
influenced by structural limitations, since the
spatial location of the bacterial population influence
the effectiveness of certain species (free swimming
and larger) (Clarholm, 1981; Vargas and Hattori,
1986; Kuikman et al., 1990). On the other hand, the
castings produced from the Lumbricus terrestris
and Aporrectodea caliginosa are favorable habitat
for protozoa, due to their higher (about 10%)
porosity than soil. Castings of Octolasion lacteum
also shows higher protozoan population than the
soil aggregates (Bonkowski, 1995). Apart from
high microbial biomass which is source of feed of
protozoans in the cast, the pore and its distribution
14
also improve the predation (Larink et al., 2001).
Oxygen concentration will also control the kind and
distribution of organisms which in turn is affected
by pore space in the castings (Elliott and Coleman,
1986).
As the changes in the pore space greatly
modify the structure of microbial community,
mineralization and immobilization of nutrients also
vary significantly as it is partially controlled by the
size, and composition of microorganisms and their
activity (Marschner and Kalbitz, 2003; Fontaine
and Barot, 2005). The mineralization of organic
matter may be limited by pore size distribution due
to the positioning of organic matter in pores which
restricts the access to the microorganisms, thereby
causing restricted predation of those
microorganisms (Elliott and Coleman, 1988;
Hassink, 1992; Ladd et al., 1993; Strong et al.,
2004). The protozoa are found in pores > 6 µm dia
in which higher C mineralization is reported than
the bacteria populated pores of < 6 µm (Killham et
al., 1993). Moreover, the pores of different size
control the exchange of water and gases, which in
turn influence the availability of organic substrates
to the organisms. For instance, in the pores of
diameter in the range of 6-30 µm, the
mineralization of C was high when these pores
were filled with water and which attributed the
availability of microbial biomass to protozoans in
these pores (Killham et al., 1993). Increasing the
proportion of water-filled porosity increases the soil
water potential which consequently increases
mineralization rates (Sommers et al., 1980) but it
may create anaerobic conditions and stimulate
denitrification when the water-filled pores are more
than 60% of total pore space (Linn and Doran,
1984; Bergstrom and Beachamp, 1993; Elliott and
de Jong, 1993).
2.2.4. Water-related properties
As the earthworms casting activity has
significant impact on the soil porous system, soil
hydraulic functions such as infiltration, erosion,
runoff, water retention and evaporation are also
strongly influenced (Lamandé et al., 2003;
Bottinelli et al., 2010). The packing voids within
cast change the infiltration rate by transporting and
retaining large amounts of water and solutes
through it (Lamandé et al., 2003), which plays
significant role in regulating soil quality. Sufficient
amount of water need to infiltrate into soil and soil
aggregates, because it may create anaerobic
conditions when it is restricted that slows down the
organic matter decomposition (Elliot et al., 1990;
Blanchart et al., 1993). Moreover, lower soil
infiltration rate is generally correlated with decline
in soil quality due to increase in runoff and soil
erosion (Piekarz and Lipiec, 2001; Bouchb and Al-
Addan, 1997).
The effects of earthworms on infiltration may
vary with ‗compacting‘ and ‗decompacting‘ species
as they produce castings of different size and
porosity (Hallaire et al., 2000). Studies conducted
on different soil systems have shown that the
castings of Pontoscolex corethrurus, a compacting
species led to significant change in structure of
surface soil. The outermost layer became highly
compacted due to numerous compact structures
which caused impermeability and reduced the
infiltration (Alegre et al., 1995; Barros et al., 1996;
Young et al., 1998; Chauvel et al., 1999; Hallaire et
al., 2000). Castings deposited by other compacting
species, Millsonia anomala has also been reported
to impede infiltration (Spain et al., 1992; Derouard
et al., 1997), in both laboratory and field studies.
The high bulk density and low mesoporosity of
compacting type castings may probably be the
reason for impeding infiltration (Blanchart et al.,
2004).
In contrast, larger size compacting type
castings produced by Amynthas khami are reported
to improve water infiltration by enhancing
roughness of the surface soil, modification in the
circulation of surface runoff and reducing the
velocity in steep-slope ecosystems of Northern
Vietnam (Jouquet et al., 2008). The differential
15
impact of castings produced by A. khami probably
due to its larger size which has large packing voids
allows a large amount of water to infiltrate.
Moreover, these castings are anchored in soil, and
are connected with below-ground galleries which
facilitate the infiltration (Jouquet et al., 2008).
However, the fresh and decompacting type castings
are highly dispersible in water, and clog the
transmission pore, thereby impeding infiltration of
water in soil (Shipitalo and Protz, 1988; Le Bayon
and Binet, 1999). A similar, impact on soil
infiltration was reported with the castings of
Aporrectodea tuberculata and Metaphire posthuma.
Although, the castings of these species initially
increased the infiltration rate due to their high
porosity, the unstable nature of this structure led to
rapid compaction of soil after rainfall or overland
flow (Ela et al., 1992; Binet and Le Bayon, 1999;
Bottinelli et al., 2010). However, balance between
the amount of cast deposited over the soil surface
and its degradation rate regulates the soil structural
porosity, which in turn influence infiltration. Soil
structural improvement would occur when the
amount of cast production predominates and the
effect would be reversed when degradation of these
structures was more (Bottinelli et al., 2010). In
addition, the ageing of castings also influence their
impact on soil. In general, dispersion of fresh
castings is high, and it becomes stable during
ageing under several drying–wetting cycles
(Decaëns et al., 1999b,c; Mariani et al., 1999). The
stable aggregates can protect from detachment by
rainfall or surface flow, and reduce the velocity of
surface runoff and as a consequence increase
infiltration.
In addition, the amount of clay present in the
castings is also one of the main soil parameter that
affect the soil surface sealing and infiltration
(Shainberg and Levy, 1996), as the increase in clay
content increases with increase in the stability of
aggregates. This is especially due to the
aggregation and bonding effect of clay (Le
Bissonnais, 1996). Thus, the stability of aggregates
against raindrop impact generally increases with an
increase in clay content (Blanchart et al., 2004).
Like the clay percentage, presence of organic
matter also affects infiltration because it is an
important stabilizing factor of aggregates (Shipitalo
and Protz, 1988). Many studies conducted in the
tropics have showed that most earthworms prefer
organic particles, depending on the species and
availability of organic content in soil (Barois et al.,
1999). As a result of difference in the behavior, the
stability of the castings they produce also vary, and
thereby its impact on soil infiltration may also be
different (Tomlin et al., 1995). The nature and
amount of exchangeable cations such as Ca2+
, Mg2+
,
K+, Na
+ and carbohydrates and lignin content of
plant debris present in the castings are reported to
influence erosion and infiltration through their
impact on dispersion and flocculation of clay (Le
Bissonnais, 1996), which may vary with different
earthworm species and substrate which is available
to them (Pop et al., 1992; Salmon and Ponge, 1999;
Oyedele et al., 2006).
2.3. Chemical properties of vermicast
2.3.1. Organic carbon
Carbon is a basic unit of all life form on
earth. Though not a nutrient element‚ it is the
building block of all organisms. Carbon is the
center for biological energy transfer within the
biosphere at landscape and ecosystem levels‚ and
within the organisms (Lavelle and Spain, 2003). In
the soil the effects of earthworm on carbon
dynamics vary, depending on the scale of space and
time that is considered (Lavelle et al., 1998). In the
short-term, earthworms enhance the carbon
mineralization of soil during their gut passage
(Blair et al., 1994); whereas, this effect is opposite
in aged castings in which carbon is protected from
further breakdown (Decaëns et al., 1999a; Kooch
and Jalilvand, 2008). Few studies revealed that
stabilized organic matter in the casts, can be
protected from microbial mineralization for many
years (Mcinerney and Bolger, 2000). The increase
in carbon stock in soil is gaining importance, as the
16
rise in CO2 and global warming are major concerns
(Bossuyt et al., 2005).
Although, castings produced by earthworms
stabilized the carbon for longer period, a
considerable amount of carbon loss through their
mucus secretion exceeds the carbon lost by their
respiration (Scheu, 1991). Intestinal mucus is
composed of amino acids of low-molecular weight
(about 200 Da) with carbohydrates and high
molecular weight glycoproteins (40000–60000 Da)
(Martin et al., 1987). Studies on 14
C and 15
N labeled
feed with Nicodrilus longus have shown that the
entire C content of the earthworm tissues could
turnover in 40 days, and a considerable portion of
this turnover was due to mucus excretion (Ferriere
and Bouche, 1985). The total C content of the
intestinal mucus of different earthworm species
ranged from 39–44 % C and 7–7.3 % N and it
seems to be similar across different species and
ecological categories (Martin et al., 1987). In the
study conducted by Scheu (1991) with geophagous
earthworm Octolasion lacteum, 63% of total C
losses (mucus excretion plus respiration) reported
was from secretion of mucus in casts and from the
body wall. This corresponds to a daily loss of 0.7%
of total C for this species, on the other hand,
respiration accounted for only 37% of total C loss.
Pontoscolex corethrurus secreted up to 50 Mg
mucus/ha in a single year in tropical pastures of
Mexico, which corresponds to 20% of the total C in
the soil (Lavelle, 1988).
The C in earthworm casts differ in form and
amount from those of the adjacent soil. Many
studies showed about 1.2 to 2 folds higher carbon
content in the castings compared to bulk soil (Lee,
1985; Shipitalo and Protz, 1988; Daniel and
Anderson, 1992; Barois et al., 1993; Zhang and
Schrader, 1993; Buck et al., 1999; Bossuyt et al.,
2005). The higher carbon content of castings are
probably due to their preferential ingestion of
organic rich residues such as leaf and root litter,
microorganisms, fecal pellets of other invertebrates,
etc. (Curry and Schmidt, 2007). It is also postulated
that the higher carbon content of castings is
probably due to the incomplete absorption of
carbon during the gut transit (Shaw and Pawluk,
1986; Daniel and Anderson, 1992). In contrast,
when the earthworms feed upon phytomass or
manure, there was drastic reduction in carbon
content in the egested castings. Aira and
Domínguez (2009) reported about 40% reduction in
C content of cow and pig manure after the transit
through the gut of Eisenia fetida. During the
digestion, absorption of C and microbial respiration
in the form of CO2 by worms reduces the C content
in the castings as compared to the feed. The
mixture of industrial lignocellulosic waste with
other organic supplements (saw dust and cow dung)
fed Perionyx sansibaricus produced castings that
exhibited a decrease in organic C content of 5.0 to
11.3% (Suthar, 2007). Similar C losses were
reported in casts of Eudrilus eugeniae fed with pig
manure (Aira et al., 2006). Contradiction in the
results is due to the preferential ingestion of high
organic matter by earthworm in field, and however,
C content of castings compared with only the bulk
soil. Therefore, earthworm castings show higher
amount of C content, still they assimilate
considerable amount C during gut transit. The gut
passage reduces by as much as 19% C in the soil
which they ingest (Edwards, 2004).
The C content of the castings vary
significantly depending on the earthworm species,
their feeding habits, particularly the amounts of
plant litter they intake (Table 2). Earthworms are
generally classified into three groups, epigeic,
anecic and endogeic (Bouché, 1977). Among them,
the epigeic and anecic species often behave as
litter-feeders, while the endogeics are geophagous,
consuming enormous quantities of soil in order to
meet their organic matter needs. Therefore, the
endogeic species are further divided into three
groups: oligohumic, mesohumic and polyhumic
(Lavelle, 1981). Based on the degree of food
selection, nutrient concentration is generally higher
in the casts in detritivorous than in geophagous
species (Buck et al., 1999). However, feeding
17
Table 2. Some of the chemical properties of castings produced by different earthworm species from different organic wastes.
Substrates Earthworm species Carbon mg g
-1 Nitrogen mg g
-1 Phosphorus mg g
-1 Potassium mg g
-1
References Cast feed Cast feed Cast feed Cast feed
Agricultural waste Eisenia foetida 35.0 106.0 8.8 1.8 1.0 0.70 2.63 0.62 Garg et al., 2006a
Buffalo dung Eisenia foetida 267.0 519.0 7.80 5.60 5.80 5.00 0.65 1.07 Garg et al., 2006b
Camel dung Eisenia foetida 407.0 463.0 5.20 4.00 3.20 3.00 0.36 0.50 Garg et al., 2006b
Coffee pulp Eudrilus eugeniae 147.0 535.0 16.6 10.3 4.10 1.30 7.00 1.20 Raphael and
Velmourougane, 2011
Coffee pulp Perionyx ceylanesis 168.0 535.0 18.6 10.3 5.10 1.30 7.80 1.20 Raphael and
Velmourougane, 2011
Couch grass (Cynodon dactylon) Eisenia fetida 342.1 497. 9 15.8 7.33 9.00 4.30 10.5 5.86 Pramanik et al., 2007
Cow dung Lampito
mauritii 268.7 298.9 7.73 5.78 2.73 2.32 6.00 5.42 Suthar, 2008
Cow dung Eisenia fetida 326.0 362.2 17.2 6.59 12.7 5.16 11.4 6.39 Pramanik et al., 2007
Cow dung Perionyx excavatus 201.6 285.9 19.3 12.8 8.13 5.72 9.51 4.93 Suthar, 2009
Cow dung Perionyx sansibaricus 200.2 285.9 20.4 12.8 8.35 5.72 9.66 4.93 Suthar, 2009
Cow dung Eisenia fetida 320.0 486.0 31.4 8.7 13.4 8.70 7.80 5.50 Yadav and Garg, 2011
Cow dung E. fetida + L. mauritii 261.7 298.9 9.18 5.78 2.77 2.32 6.15 5.42 Suthar, 2008
Dairy sludge + cattle manure Eisenia andrei 225.0 303.0 17.0 11.0 7.70 7.30 7.60 25.0 Elvira et al., 1998
Digestate from biogas plant Perionyx excavatus 285.0 364.0 40.0 18.0 14.0 9.00 10.4 6.50 Rajpal et al., 2014
Digestate from biogas plant Perionyx sansibaricus 276.0 364.0 40.0 18.0 13.0 9.00 10.0 6.50 Rajpal et al., 2014
Donkey dung Eisenia foetida 381.0 485.0 6.80 5.00 5.50 5.00 7.30 13.1 Garg et al., 2006b
Duckweed residues Eisenia fetida 261.5 445.8 13.1 6.24 9.0 3.96 10.5 5.72 Pramanik et al., 2007
Filter mud + saw dust Eisenia foetida 349.0 400.3 22.8 16.0 24.7 12.6 9.60 4.40 Khwairakpam and
Bhargava, 2009
Filter mud + saw dust Perionyx excavatus 357.0 400.3 24.4 16.0 22.9 12.6 7.50 4.40 Khwairakpam and
Bhargava, 2009
Filter mud + saw dust Eudrilus eugeniae 363.0 400.3 28.0 16.0 24.4 12.6 8.40 4.40 Khwairakpam and
Bhargava, 2009
Food industry sludge + poultry
droppings + cow dung Eisenia fetida 330.0 400.0 29.4 18.0 11.9 8.90 5.50 4.00
Yadav and Garg, 2011
Goat dung Eisenia foetida 231.0 438.0 8.90 4.70 4.70 3.70 3.40 7.20 Garg et al., 2006b
Guar gum industrial waste + cow
dung + saw dust Perionyx sansibaricus 431.7 481.9 18.3 15.4 3.90 2.50 13.2 11.5 Suthar, 2007
Horse dung Eisenia foetida 216.0 484.0 7.70 3.50 9.60 7.00 3.80 7.80 Garg et al., 2006b
Institutional waste Eisenia foetida 32.0 55.0 7.30 1.40 0.87 0.40 0.72 0.35 Garg et al., 2006a
18
Substrates Earthworm species Carbon mg g
-1 Nitrogen mg g
-1 Phosphorus mg g
-1 Potassium mg g
-1
References
Cast feed Cast feed Cast feed Cast feed
Kitchen waste Eisenia foetida 33.0 73.0 11.0 2.50 1.80 1.30 4.36 0.87 Garg et al., 2006a
Municipal solid wastes Eisenia fetida 242.6 200.3 7.62 3.74 5.10 3.52 7.00 4.28 Pramanik et al., 2007
Paper mill sludge + cattle
manure Eisenia andrei 151.0 266.0 12.0 11.0 5.90 3.90 7.60 23.0 Elvira et al., 1998
Poultry droppings + cow dung Eisenia fetida 350.0 420.0 28.5 11.9 12.9 9.00 6.10 4.30 Yadav and Garg, 2011
Press mud + cow dung Perionyx ceylanensis 290.0 544.1 16.3 9.40 23.8 15.0 31.3 21.2 Prakash and Karmegam,
2010
Sewage sludge Eisenia fetida 219.3 267.6 30.0 26.8 47.7 43.7 5.59 5.10 Suthar, 2010
Sewage sludge+ sugarcane trash Eisenia fetida 218.5 320.4 29.3 23.5 28.9 25.4 7.10 6.30 Suthar, 2010
Sheep dung Eisenia foetida 212.0 323.0 7.80 3.70 5.10 3.10 4.50 7.00 Garg et al., 2006b
Spent mushroom compost E. foetida + E. andrei 38.4 37.9 29.5 28.3 22.9 12.7 14.6 8.70 Tajbakhsh et al., 2008
Spent mushroom compost +
fruits and vegetables residues E. foetida + E. andrei 45.3 43.9 25.8 21.1 44.9 7.50 34.1 9.14 Tajbakhsh et al., 2008
Spent mushroom compost +
pomegranate residues E. foetida + E. andrei 43.3 46.5 28.9 22.2 34.3 7.60 29.5 7.19 Tajbakhsh et al., 2008
Spent mushroom compost +
potato residues E. foetida + E. andrei 81.1 45.9 21.4 22.1 36.8 7.95 81.9 13.7 Tajbakhsh et al., 2008
Spent mushroom compost +
stump residues E. foetida + E. andrei 13.4 28.9 19.5 14.6 24.1 9.05 27.4 10.6 Tajbakhsh et al., 2008
Spent mushroom compost +
tomato residues E. foetida + E. andrei 33.4 46.5 4.80 34.9 35.6 8.95 37.5 5.54 Tajbakhsh et al., 2008
Textile industry fibre waste Eisenia foetida 38.0 64.0 0.52 1.00 0.60 0.26 2.47 0.82 Garg et al., 2006a
Textile industry sludge Eisenia foetida 34.4 51.0 7.00 1.20 0.26 0.04 0.43 0.30 Garg et al., 2006a
Textile mill sludge+ cow dung Eisenia foetida 170.0 297.0 9.20 3.50 4.10 3.70 2.30 4.40 Kaushik and Garg, 2003
Winery industry waste Eisenia andrei 418.0 546.0 14.0 15.6 4.97 2.23 18.2 20.8 Nogales et al., 2005
19
behavior of earthworms also vary with different soil
type where they inhabit. Barois and Lavelle (1986)
demonstrated that the tropical species Pontoscolex
corethrurus was able to selectively ingest either
small mineral particles or large organic debris,
depending on the type of soil. Selection was made
on aggregates rather than primary particles.
However, some earthworms may selectively ingest
coarser particles than the average in soils with very
high clay contents. As a consequence, the C content
of castings also varies with species involved and
place they inhabit.
Moreover, the digestive systems of
earthworms of different ecological category have
different assimilation efficiencies, which also could
be the reason for differential carbon content of their
castings. Studies of Jegou et al. (1998) had shown
castings of Eisenia andrei (epigeic), Lumbricus
terrestris (epi-anecic) are enriched with organic
matter compared with bulk soil, whereas, those of
Aporrectodea giardi (anecic) and Aporrectodea
caliginosa (endogeic) contains lesser C than their
surrounding soil. The castings of epigeic and
epianecic species contained more than 50% litter C
(13
C labeled), while the anecic and endogeic were
of soil. Studies conducted in absence of leaf litter
showed that the C content of castings from the
geophagous Allolobophora chlorotica exceeded
that of Lumbricus rubellus, Lumbricus terrestris
and Octolasion cyaneum in soil without leaf litter
(Bishop et al., 2008). The phytophagous species,
which ingest organic material with more readily
degradable C, had greater assimilation capacities,
which resulted in lesser carbon content in cast
(Edwards and Bohlen, 1996). The feed quality such
as C/N ration, fibre content, and other secondary
metabolite are also reported to influence the C
states of castings (Mangold, 1951; Buck et al.,
1999).
In the earthworm castings, organic carbon
accumulates in different soil size fractions
(Guggenberger et al., 1996). However, distribution
pattern of C with all size fraction vary with soil
organic matter which they ingest and species of
earthworm. Organic matter associated with soil of
different size fraction (sand, silt and clay)
significantly exhibited different properties in terms
of organic matter turnover (Tiessen and Stewart,
1983; Anderson and Paul, 1984). Sand-associated
organic matter is important in short-term turnover,
clay-bound organic matter dominates the medium-
term turnover, whereas silt-bound organic matter
take part in long-term turnover. Therefore, particle-
size separation aids in estimating labile and passive
organic carbon fractions (Christensen, 1992).
In general, the clay bound C content seem to
be higher in the casts (Scullion and Malik, 2000;
Marhan et al., 2007). The organic matter bound to
clay is known to be stable and the formation of pool
of clay bound carbon is slow (Hassink and
Whitmore, 1997). Earthworms may enhance the
grinding and mixing organic matter with mineral
soil particles of the clay fraction thereby increasing
stabilized organic matter in soil (Oades, 1988). This
shows that clay bound organic matter is influenced
by gut transit. The C gains also high in fine-sand
sized fraction, almost twice as much C in the casts
as in the surrounding soil reported by Zhang et al.
(2003). Earthworm casts exhibited the lowest C
concentration in the sand-sized separates, but
compared with the sand-sized separates of the
surrounding soil they were much less depleted in
organic C. After gut passage a pronounced
depletion of C associated with sand and an
enrichment of silt- and especially clay-bound C is
reported. Hence, earthworm activity resulted in a
pronounced redistribution of C within size
separates (Guggenberger et al., 1996). In the
endogeic earthworm, Millsonia anomala about 25%
of C content with coarse fractions reduced during
gut transit, while C in the finer fractions tended to
increase. Similar changes were also encountered
with another species Polypheretima elongata in a
Martiniquan vertisol (Lavelle and Spain, 2003).
20
2.3.2. Macronutrients (primary)
Nitrogen – Nitrogen is critical to all the life
forms and is an important constituent of many
biologically-active compounds. It plays a
significant role in all the major category of
biological function in both animal and plant
(Lavelle and Spain, 2003). In terrestrial ecosystem
earthworms significantly influence the nitrogen
through modification of the physical, chemical and
biological properties of soil (Lee, 1985; Edwards
and Bohlen, 1996). In addition, earthworms
contribute to the mineral nitrogen pool of soil
directly due to excretion of nitrogenous compounds
in their urine and mucus (Whalen et al., 2000).
Castings they produce are reported to contain
higher total and extractable nutrients than the bulk
soil (Edwards and Bohlen, 1996), but the nitrogen
content of castings produced under field condition
was not always richer than bulk soil. The total
nitrogen content of casts is related to earthworm
diet and so may be similar to or greater than the
values reported for bulk soil, depending on nitrogen
in organic substrates ingested by earthworms (Syers
et al., 1979; Scheu, 1987; Buck et al., 1999;
Perreault et al., 2007; Kizilkaya, 2008). Flegel and
Schrader (2000) observed that the total nitrogen of
castings produced by Dendrobaena octaedra was
about twofold higher with dandelion as food,
compared to those fed with larch. In addition,
earthworm feeding habit also significantly
influenced its nitrogen value. For instance, the
castings of Octolasian tyrtaeum were of higher
mineral nitrogen with dry vetch as feed in
comparison to green vetch (Parkin and Berry,
1994).
Earthworm‘s castings are widely reported to
contain elevated amount of mineral nitrogen,
especially for the ammonium content in relation to
surrounding soil (Parkin and Berry, 1994; Decaëns
et al., 1999a; Whalen et al., 2000; Aira et al., 2003;
Bityutskii et al., 2012; Clause et al., 2013). High
mineral nitrogen content of fresh castings is
probably due to their preferential feeding of
nitrogen rich substrates and high rate of
mineralization of organic matter during gut transit
through their ―priming effect‖ (Lavelle et al.,
1995). Microorganisms ingested along with the soil
are stimulated by earthworm inner living conditions
and increase the mineralization of organic matter in
the earthworm gut (Lavelle et al., 1995; Aira et al.,
2003; Chapuis-Lardy et al., 2010), and this
continues for several hours in fresh casts (Barois
and Lavelle, 1986). In addition, the low
assimilation efficiencies of earthworms and urine in
the hind part of the digestive tract also could be the
reason for high ammonia content of vermicast
(Syers and Springett, 1983; Anderson et al., 1983;
Parkin and Berry, 1994; Aira et al., 2003).
In fresh castings, about 96% of mineral
nitrogen was present as ammonium (Parle, 1963). A
fraction (6%) of the non-plant available N ingested
by Allolobophora caliginosa (Savigny) was
excreted in plant available form (Syers et al., 1979).
The ammonium content of castings progressively
decreases with age (Decaens et al., 1999a;
Chevallier et al., 2006). The reason for the decrease
may be due to the NO3- production in fresh casts as
a result of nitrification (Scheu, 1987; Lavelle and
Martin, 1992). As the days progressed, the NO3-
content of vermicast also largely decreased
(Decaëns et al., 1999a), obviously due to root up-
take, immobilization in the soil microbial biomass,
denitrification processes or losses by leaching
(Syers et al., 1979; Elliott et al., 1990; Lavelle and
Martin, 1992).
Higher mineral nitrogen content and water-
filled pore space of vermicast and anaerobic
condition of the earthworm gut lead to increase in
the denitrification rate in the fresh cast (Elliott et
al., 1990; Knight et al., 1992; Granli and Bøckman,
1994; Smith et al., 1998), which is much higher
than the adjacent soil (Svensson et al., 1986; Elliot
et al., 1990; Parkin and Berry, 1994). Thus, gaseous
losses of nitrogen from vermicast continuous till the
production of nitrate ceased by intense nitrification.
Kharin and Kurakov (2009) observed about 1.5
21
times higher denitrification rate in fresh castings of
Aporrectodea caliginosa than that in the soil, and it
reduced by 30% and became closer to the activity
of denitrification in the soil after 12 days. In the
case of castings produced by Lumbricus terrestris,
about 2.5 times higher N2O emission is reported
than surrounding soil (Svensson et al., 1986).
Denitrification in cast may vary with different feed.
Parkin and Berry (1994) observed that castings
from worms allowed to feed on hairy vetch showed
higher denitrification rates than the castings
produced by earthworms which were manure fed or
not supplemented with organic material. The
denitrification rates of Octolasian tyrtaeum and
Aporrectodea tuberculata casts from vetch were
significantly higher than those from horse manure
or only soil. A few studies have shown that
earthworms make a substantial contribution to total
N2O emitted (30–56%) from the soils they inhabit
(Knight et al., 1992; Karsten and Drake, 1997;
Matthies et al., 1999; Borken et al., 2000).
Phosphorus – Phosphorus is one of a major
elements required by all the life form. It is an
essential component of ATP, ADP,
phosphoproteins, phospholipids of nucleic acids
and coenzymes. Phosphorus deficiencies lead to
suppression of many biological activities, so that
phosphalic fertilizers are widely used in agricultural
systems to improve the productivity of crops.
Phosphorus is one of the element which does not
have gaseous phase in its biogeochemical cycle.
Therefore phosphorus is largely cycled in soil by (i)
organic matter decomposition‚ (ii) turnover of the
microbial biomass‚ (iii) weathering of the parent
material and‚ (iv) in agro-ecosystems‚ as fertilizers
(Lavelle and Spain, 2003). In soil, the phosphorus
exists in different forms and concentrations.
However, most of phosphorus are associated with
aluminum‚ iron, calcium, manganese and other
carbonates, and became unavailable to plants. In
general, the availability of phosphorus in soil
mainly relies on pH and mineral composition of
soil, and few forms of phosphorus such as
dihydrogen phosphate and hydrogen phosphate that
can be easily utilized by plants (Lavelle and Spain,
2003; Chapuis-Lardy et al., 2009).
Earthworms are one of the important soil
organisms that can influence the availability of
phosphorus in soil. Through ingestion of particles
rich in P, the earthworms accumulate P in the casts.
They modify the proportion and stability of
different forms of P in castings (Brossard et al.,
1996; Kuczak et al., 2006) of species from tropical
and temperate regions (Sharpley and Syers, 1976;
Barois et al., 1987; Lavelle and Martin, 1992;
Lavelle et al., 1992; López-Hernández et al., 1993;
Scheu, 1987). Higher concentration of P in castings
may be probably due to (i) higher pH of the
intestinal gut contents (6.8, 6.0 and 4.6 for the
anterior and posterior parts and soil, respectively)
(Barois and Lavelle, 1986); (ii) a huge amounts of
mucus in the earthworm gut which is a mixture of a
glycoprotein, amino acids and sugars (Lavelle et
at., 1983; Martin et al., 1987) can inhibit and
compete for orthophosphate sorbing places by
carboxyl groups of carbohydrate compounds they
contain (López-Hernández et al., 1993), and (iii)
production of organic acids during digestion due to
increase in microbial activity which may compete
for P-sorbing sites and increase P solubility (Earl et
al., 1979; López-Hernández et al., 1993).
Moreover, enhanced phosphatase and microbial
activities in the biogenic structure of earthworm
and its surrounding increases availability of P
(organic and inorganic) (Sharpley and Syers, 1976;
Mulongoy and Bedoret, 1989; Edwards, 1998;
Richardson and Simpson, 2011).
The impact of earthworms on the availability
of P may be varying with different species, because
digestive abilities and gut microorganisms have
been reported to differ among species (Lattaud et
al., 1998). The casting of an epi-endogeic
earthworm, Lumbricus rubellus has been reported
to produce casts of higher exchangeable P, then
Lumbricus terrestris, an anecic species (Suárez et
al., 2003). The surface castings of Allolobophora
caliginosa is reported to contain about fourfold
22
more plant-available inorganic P and twice as much
plant-available organic P as the underling soil
(Sharpley and Syers, 1977), and those of
Pontoscolex corethrurus showed 2.7-fold increases
in water-soluble P after gut passage (López-
Hernández et al., 1993). However, increment in
available P was maintained for only few days, and
it decreased rapidly with age. Reduction in
available P was reported on fourth day and three
weeks after castings deposited by Pontoscolex
corethrurus (Lopez-Hernandez et al., 1993) and
Lumbricus terrestris, respectively (Basker et al.,
1993). The availability of high carbon and nitrogen
content in fresh cast increases the phosphorus
demand and subsequently immobilization P by
microorganisms may probably be the reason for P
decline.
Potassium – All the living organisms require
potassium, usually in relatively large amounts as
that of other major nutrients. It plays major role in
regulating many physiological and biochemical
processes in plants, such as pH stabilization,
osmoregulation, membrane transport and enzyme
activation and; thereby it influences photosynthesis,
protein synthesis and many other cellular and
physiological processes (Marschner, 1995). In soil
system, potassium is largely cycled by weathering
of primary minerals such as feldspars, micas, etc.
and mineralization of organic matter (Lavelle and
Spain, 2003). There are plenty of evidences on the
favorable effect of earthworms on soil potassium
status, through their borrowing and castings
activities. Many have reported a nominal increase
in total potassium and a significant increment in its
soluble form in vermicast compared to earthworm
feed or surrounding soil. However, the status of
potassium in castings may vary with different
earthworm species and feed they ingest.
Total potassium content in casting of Eisenia
fetida from cow dung, grass, aquatic weeds and
municipal solid waste increased to the extent of
28% compared to the raw feed (Pramanik et al.,
2007). Increase in potassium content is also
reported with castings produced from agro-
residues, kitchen waste, industrial and institutional
wastes including textile industry fibres and sludge
by the same earthworm species (Garg et al.,
2006a); whereas, castings were of lower potassium
content when they were fed with coffee pulp waste
(Orozco et al., 1996). However, castings from the
same coffee pulp waste showed higher potassium
content when it was produced by Eudrilus eugeniae
and Perionyx ceylanesis (Raphael and
Velmourougane, 2011). Increase in total potassium
content has also been reported with castings of
Eisenia fetida, Perionyx excavatus and Perionyx
sansibaricus with sewage sludge and anaerobic
digestate as feed, respectively (Delgado et al.,
1995; Rajpal et al., 2014). While casting of Eisenia
foetida and Eisenia andrei from excreta of different
livestock (cow, buffalo, horse, donkey, sheep, goat
and camel) and industrial sludges (paper mill, dairy
and leather industries), had lower potassium content
than the initial substrate (Elvira et al., 1998; Garg et
al., 2006b; Ravindran et al., 2008). These
differences in response of earthworms depends on
the chemical nature of the substrate and difference
in digestive ability of earthworm species.
In general, the increase in total potassium
content in castings probably due to ‗concentrative
effect‘, in which the loss of organic matter as CO2,
N2 or N2O during the gut transit, concentrated the
potassium content in ejected castings. Few studies
have also reported lower potassium in castings,
which might be caused by excess potassium uptake
by worms or leaching of soluble elements by excess
water that drained through the feed and vermicast
(Garg et al., 2006b; Ravindran et al., 2008;
Tajbakhsh et al., 2008). Leachate collected during
vermicomposting process is reported to contain
potassium in a significantly high quantity (Benitez
et al., 1999).
Many have reported significantly higher
exchangeable potassium content in castings in
comparison to the raw substrate or adjacent soil.
Castings produced in field had shown two to
23
threefold higher exchangeable K than the
surrounding soil (Tiwari et al., 1989; Bezborodov
and Khalbayeva, 1990; Hullegalle and Ezumah,
1991; Jouquet et al., 2008; Pommeresche et al.,
2009), however, it may vary with soil type and
species of worm which produced the castings
(Basker et al., 1994). For instance, in sandy soil,
the castings produced by Lumbricus rubellus had
higher K content than the Allolobophora
caliginosa; whereas, the castings produced in the
silty-clay soil had lower K in compared with
surrounding soil (Basker et al., 1994). Clause et al.
(2013) reported K content was not different
between soil and castings of Aporrectodea rosea
and Allolobophora chlorotica from Rendzic
Leptosol soil, but there was an increase in K
content with Lumbricus terrestris castings in the
Histosol. Laboratory based studies on Lampito
mauritii and Eisenia fetida with cow dung as feed,
showed 9.8–13.5% increase in exchangeable
potassium content in vermicast (Suthar, 2008).
Suthar (2007) reported that the exchangeable
potassium content in castings of Perionyx
sansibaricus had increased to the extent of about
20%, with guar gum industrial wastes. Similarly,
the castings of Perionyx excavatus and Perionyx
sansibaricus from cattle waste solids had twice the
concentration of exchangeable potassium than the
initial substrate (Suthar, 2009). Increase of
exchangeable potassium content in the castings
may be due to the release of K from the non-
exchangeable potassium pool in soil and organic
material during the gut transit (Basker et al., 1992,
1993) through, (i) secretion of acids by gut
associated microorganisms, which solubilize the
insoluble potassium in feed, and (ii) the enhanced
microbial activity in earthworm gut mineralizes the
organic bond potassium in their feed. However, the
release of K from the non-exchangeable pool is a
complex phenomenon which is controlled partly by
the level of exchangeable K in the ingested feed.
Higher exchangeable K in feed can inhibit the
release of further exchangeable K from non-
exchangeable pool (Feigenbaum et al., 1981;
Basker et al., 1994).
2.3.3. Macronutrients (secondary)
Calcium – Calcium is an essential element of
the cell walls of higher plants where it occurs along
with pectin. It plays major role in various
physiological processes in plants. In soil, calcium
strongly influences the pH status, and structural
stability of soil aggregates and so on (Lavelle and
Spain, 2003). Earthworms significantly influence
the soil Ca content by their castings activity. Many
species of earthworms produce calcite granules in
specialized glands called as calciferous gland,
which are three pairs of swellings away from the
oesophagus on the 10-12 segments (Canti, 1998;
Canti and Piearce, 2003; Clause et al., 2013;
Versteegh et al., 2014). These calcite granules are
produced by fixation of CO2 with Ca from their diet
under the catalytic reaction of carbonic anhydrase
present in calciferous glands, and these granules are
ejected along with their castings (Padmavathiamma
et al., 2008). Major portion of these granules
consists of calcite, and a small quantity of
amorphous calcium carbonate, aragonite and
vaterite also present in it (Gago-Duport et al., 2008;
Lee et al., 2008; Fraser et al., 2011; Brinza et al.,
2013). The production of calcite granules is likely
related to functions of regulating pH and the
concentration of CO2 in body fluids, and regulating
Ca2+
and other potentially toxic cations in their feed
(Robertson, 1936; Crang et al., 1968; Piearce,
1972; Bal, 1977; Becze- Deák et al., 1997).
The calcite granules production varies
significantly with different earthworm species and
the feed they ingest. Lumbricus terrestris is a major
calcite producing species in temperate soils.
Production rates are between 0.8 to 2.9 mg worm-1
day-1
(Canti, 2007; Lambkin et al., 2011; Versteegh
et al., 2013). Mršić (1997) and Udovic et al. (2007)
reported that Eisenia fetida, an epigeic earthworm
have active calciferous glands, while endogeic
earthworm, Octolasion tyrtaeum possess
calciferous glands that are intermediate in
complexity and activity. Increase in calcite granules
with elevated temperatures and atmospheric CO2
24
level, were also reported in few studies (Kühle,
1980; Versteegh et al., 2014). These granules are
very stable in nature, which are expected to survive
in soils for more than 300,000 years (Lambkin et
al., 2011; Versteegh et al., 2014).
In general, the vermicast are rich in calcium
content due to their calciferous glands activity and
higher mineralization capacity. Tajbakhsh et al.
(2008) reported about two to threefold increase of
calcium content in the castings of Eisenia foetida
and Eisenia andrei than the initial substrates of
compost produced from spent mushroom. The
castings of Eisenia fetida from sewage sludge,
coffee pulp waste and mixture of solid textile mill
sludge and poultry droppings also had higher Ca
content than its initial feeds (Delgado et al., 1995;
Orozco et al., 1996; Garg and Kaushik, 2005).
However, Elvira et al. (1998) and Kaushik and
Garg (2003) have reported a decrease in calcium
content in castings after processing of sludge of
paper pulp-mill, and solid textile mill with cow
dung by Eisenia andrei and Eisenia foetida,
respectively. The differences in the results may be
due to leaching of calcium by the excess water that
passed through the feed mixtures.
Calcium assimilation in the castings has
reported to be influenced by the soil type and the
concentration of calcium it contains. Castings
produced by Hyperiodrilus africanus had shown
twice the concentration exchangeable Ca than the A
horizons, and had about three times higher
exchangeable Ca than the B horizons in Lixisol and
Luvisol soils (Oyedele et al., 2006). Similarly, the
castings of Aporrectode caliginosa generated in
loam soil also showed more than two and threefold
higher Ca content compared to the soil of Ap and
Bt horizons, respectively (Schrader and Zhang,
1997). However, castings of same species had
lower Ca content than the P horizon of the clay soil.
Although, in the clay soil, Lumbricus terrestris
produced castings from the P horizon contained
lower exchangeable Ca than the soil in which they
produced, it was about double quantity of Ca found
in the castings of Ap horizon (Schrader and Zhang,
1997). While the Ca content of castings produced
by the Lumbricus terrestris, Allolobophora
chlorotica and Aporrectodea rosea in the Rendosol,
Luvisol and Histosol type soil were not
significantly different from their surrounding soil
(Clause et al., 2013). This could be attributed to the
absence or the very high content of CaCO3 in the
Luvisol and Rendosol soils, respectively which
might have masked impact of earthworms. The
availability of Ca is very scare in the few soil
systems like Apalachicola national forest, in which
the earthworm species Diplocardia mississippiensis
and Arctiostrotus sp. are believed to ingest fungi to
derive the oxalates bound calcium they contained
(Spiers et al., 1986; Lachnicht and Hendrix, 2001).
Ca present in the castings has various impact
on soil: (i) Ca2+
cation improve soil structure by
bridging with clay and soil organic matter, and
prevent the clay dispersion and the associated
disruption of aggregates by replacing Na+ and Mg
2+
in clay and aggregates, thereby enhancing the
stability of aggregates (Armstrong and Tanton,
1992; Zhang and Norton, 2002; Bronick and Lal,
2005), (ii) Ca preserve the flocculated structure of
the clay in soil surface aggregates by neutralizing
acids produced by fungi, microbes and roots
(Bullinger-Weber et al., 2007), (iii) Ca present in
the castings neutralize the acid soil by acting as a
buffering agent (Sanyal, 1991), (iv) calcium
influence the availability and solubility of other
plant nutrients present in the castings (Edwards and
Lofty, 1977), and (v) buffering capacity of Ca
present in the castings is also reported to enhance
the nitrifying bacterial populations in the
drilosphere (Parkin and Berry, 1999).
Magnesium – It plays important role in
regulating intra-cellular osmotic potential, protein
synthesis and it is an essential component in
chlorophyll synthesis. Magnesium is also
responsible for activating a wide range of enzymes
and, has a structural role in stabilizing the cell wall
(Marschner, 1995). Earthworm castings have been
25
found to contain elevated amounts of Mg relative to
adjacent soil (Tiwari et al., 1989; Parkin and Berry,
1994). The Mg content of castings produced by
Eudrilus eugeniae had increased to the extent of
40% compared to the initial substrates of rice straw
and rice husk amended with different proportion of
cow dung. Moreover, in these castings, Mg content
increased with increasing proportion of CD with
these feed (Shak et al., 2014). Similar increase in
Mg were also observed with castings of Eudrilus
eugeniae and Perionyx ceylanensis with mixture of
coffee pulp, cow dung and farm yard manure as
feed (Raphael and Velmourougane, 2011).
Asawalam (2006) reported about twice the
concentration of Mg in the castings than its
surrounding soil. Prakash and Karmegam (2010)
also reported about twofold increases in Mg content
of casings produced by Perionyx ceylanensis with
pressmud as feed. Castings of Eisenia sp. showed
20% increase in Mg content with apple pomace
waste (Hanc and Chadimova, 2014). While Eisenia
foetida and Eisenia andrei castings had about
threefold higher concentration of Mg with spent
mushroom compost (Tajbakhsh et al., 2008).
Pattnaik and Reddy (2010) also reported about
threefold increase in Mg content of castings
produced by Eudrilus eugeniae, Eisenia fetida and
Perionyx excavatus with vegetable market waste
and floral waste. The Mg content of castings also
reported to vary with different type of soil and its
layer (Basker et al., 1993; Pattnaik and Reddy,
2010). Suthar (2010) postulated that higher activity
of fungal and microalgae in fresh castings attributed
to the increase in concentration of Mg in castings.
Loss of organic matter during the gut transit also
may be leading to higher concentration of Mg in
castings (Wani et al., 2013).
Sulfur – Sulfur plays vital roles in a number
of metabolic pathways. It is a component of the
essential amino acids cystine‚ cysteine and
methionine and thus forms a structural part of tissue
proteins and enzymes important in photosynthesis
and nitrogen fixation. In most soils‚ sulfur occurs
largely in organic form and often associated with
the organic matter of soil and decaying plant
residues. This organic bound sulfur gets
mineralized to sulfate by the soil biota which can be
easily taken up by plants (Lavelle and Spain, 2003).
Among the different soil biota, earthworms
significantly influence the availability of sulfur in
the soil. Castings produced by them are reported to
contain higher amount of available sulfur than the
feed they ingest. Prakash and Karmegam (2010)
reported about twice the concentration of sulfate in
the castings of Perionyx ceylanensis with mixture
of pressmud and cow dung as feed. While sulfate
content of castings produced by Dravida
assamensis were sevenfold higher than the
surrounding soil (Ganeshamurthy et al., 1998).
Studies of Singh and Suthar (2012) also reported
higher sulfate content in castings of Eisenia fetida
with cow dung amended pharmaceutical industrial
waste, in which increasing proportion of cow dung
from 0 to 100% in initial feed mixture increased the
sulfate content about 50 to 270%, respectively in
the vermicast. However, the castings of similar
species with mixture of aerobic and anaerobic
biological sludges as feed showed lower sulfate
content than its initial feed mixture (Masciandaro et
al., 2000). Sangwan et al. (2010) reported no
difference between the vermicast generated from
cow dung and filter cake mixed with horse dung by
using Eisenia fetida. Increasing concentration of
available sulfur in the castings is probably due to
the mineralization of organic bound sulfur by the
enhanced microbial and enzyme activity in the
earthworm gut (Ganeshamurthy et al., 1998).
Leaching or gaseous loss of sulfur during the
bioconversion process might be the cause of sulfate
reduction observed in few studies.
2.3.4. Micronutrients
Micronutrients such as boron, chloride,
copper, iron, manganese, molybdenum, zinc and
nickel are important for all the plants (Brady and
Weil, 1999). Few other nutrients under this
category (i.e. sodium, cobalt, aluminum, etc.) are
considered to be essential for the plants growth,
26
however, certain species require these nutrients
only under specific environmental conditions. For
instance, plants that use C4 pathway of
photosynthesis and crassulacean acid metabolism
(CAM) require sodium (Welch and Shuman, 1995).
Leguminous plants require cobalt for symbiotic
fixation of nitrogen by Rhizobium bacteria in their
root nodules (O'hara et al., 1988). Availability and
mobility of these micronutrients in the soil is
largely influenced by earthworms‘ burrowing and
feeding activities. Castings they produced have
been reported to contain elevated amount of
micronutrients and which is largely assessed by the
nutrient levels of feed they ingest (Table 3). For
instance, castings of Aporrectodea caliginosa
showed higher availability of Fe, Zn and Mn in the
dried rye straw (Cecale cereale L.) treated soil than
those of clover aboveground parts (Trifolium
pratense L.) treated one (Bityutskii et al., 2012).
The majority of studies have shown that
increase in concentration of micronutrients in
vermicast in comparison to their surrounding soil or
feed they ingest (Ireland, 1975; Kizilkaya, 2004;
Wen et al., 2004, 2006; Asawalam and Johnson,
2007; Dandan et al., 2007; Udovic and Lestan,
2007; Udovic et al., 2007; Karmegam and Daniel,
2009). An elevated concentration of molybdenum
was recorded in castings of Aporrectodea
caliginosa which result in enhanced nitrogen
assimilation and fixation activities in maize
seedling through activation of Mo-depending
enzymes nitrate reductase and nitogenase,
respectively (Tomati et al., 1996). Lukkari et al.
(2006) also reported higher extractable metals in
the castings of Aporrectodea caliginosa tuberculata
than bulk soil. Oyedele et al. (2006) and Bartz et al.
(2010) also reported increase in Fe availability in
soil after processed by Pontoscolex corethrurus.
Increase in Fe content of castings attributed to
increase in amorphous form of Fe to available Fe
during the gut transit (Bartz et al., 2010). Oyedele
et al. (2006) postulated that higher contents of
extractable Fe and Al in casts are probably due to
transformation of crystalline fractions ingested soil
particles by the gizzard‘s crushing and mixing
activities. In addition, reduction of Mn and Fe in
the soils is facilitated by earthworm‘s anaerobic gut
passage due to lower redox potential of these
elements (Duarte et al., 2012).
The Fe, Mn and Zn content of Eisenia fetida
castings from municipal solid waste had increased
to the extent of 15, 32 and 15%, respectively
(Suthar et al., 2014). Bityutskii and Kaidun (2008)
have reported about five to tenfold increase in
water-soluble Fe and Mn in the fresh casting of
Eisenia fetida, Aporrectodea caliginosa, and
Lumbricus terrestris, and the solubility of these
nutrients increased further in 9 days older castings.
Among these earthworm species, Eisenia fetida had
showed lowest nutrients content in both fresh and
old castings. In sewage sludge amended soil,
Lumbricus terrestris produced castings of about
two and sevenfold higher Zn and Cu content than
its surrounding soil (Kızılkaya, 2004). Similarly,
castings of Pontoscolex corethrurus in the soil
contaminated with Pb mining activities had shown
elevated amount of Fe, Mn and Al oxides than its
surrounding soil (Duarte et al., 2012). Increase in
Fe, Z, Mn and Cu have also been reported with
vermicast from manure amended sludges from food
industries, dairy, paper mill, fly ash and winery
industries (Elvira et al.,1998; Nogales et al., 2005;
Venkatesh and Eevera, 2008; Yadav and Garg,
2011). Increase in concentration of micronutrients
in the vermicast could be due to (i) breakdown of
organic matter by the stimulated enzymes and
microbial activities in the earthworm gut, and
release of organically bound nutrients it contains
(Rada et al., 1996), and (ii) production of soluble
organics such as humic acids and fulvic acid during
the gut passage elevates the micronutrients
availability by the formation of water-soluble
complexes (Halim et al., 2003; Evangelou et al.,
2004).
Few studies have also reported lower
concentration of micronutrients in vermicast
compared to the initial feed or soil. For example,
27
Table 3. Some of the micronutrient content of castings produced by the different earthworm species from the different organic wastes.
Substrates Earthworm species Zn mg kg
-1 Cu mg kg-1 Fe mg kg
-1 Mn mg kg-1
References Cast feed Cast feed Cast feed Cast feed
Cattle dung Eisenia fetida 11.5 7.55 9.0 3 2.04 590.0 498.4 38.0 25.0 Bhat et al., 2013
Cattle dung Eisenia andrei 155.0 108.0 36.0 34.0 9600 6100 240.0 198.0 Elvira et al., 1998
Cow dung Eisenia fetida 155.0 117.3 - - 1368 1261 569.3 546.7 Suthar et al., 2014
Cow dung Eisenia fetida 193.0 145.0 52.6 32.4 2280 1810 - - Yadav and Garg, 2011
Dairy sludge + cattle dung Eisenia andrei 198.0 198.0 43.0 39.0 9300 7400 218.0 298.0 Elvira et al., 1998
Distillery sludge + cow dung Eisenia fetida 246.0 368.2 30.5 41.1 377.0 457.9 238.3 295.8 Suthar, 2008
Dyeing sludge Eisenia fetida 38.8 37.6 15.1 14.1 1600 1510 295.1 235.1 Bhat et al., 2013
Dyeing sludge + cattle dung Eisenia fetida 29.0 26.0 15.0 11.0 1456 1380 240.1 184.1 Bhat et al., 2013
Empty fruit bunch of oil palm Eudrilus euginae 10.6 2.82 9.59 2.18 9.29 1.62 18.75 16.8 Hayawin et al., 2010
Fly ash + cow dung Eudrilus eugeniae 14.0 8.60 11.8 1.20 64.9 27.5 29.6 12.9 Venkatesh and Eevera, 2007
Food industry sludge + poultry
droppings + cow dung Eisenia fetida 805.0 475.0 77.8 59.8 1400 1280 - - Yadav and Garg, 2011
Horse dung Eisenia fetida 917.0 870.0 387.0 153.0 25791 18630 1671 1121 Sangwan et al., 2008a
Itchgrass + cow dung Lampito mauritii 75.2 84.6 13.8 13.2 2300 2100 8300 9500 Karmegam and Daniel, 2009
Itchgrass + cow dung Perionyx ceylanensis 77.0 84.6 13.4 13.2 2400 2100 8700 9600 Karmegam and Daniel, 2009
Leaf litter of Indian mast tree +
cow dung Lampito mauritii 38.4 43.0 34.4 31.2 2100 1300 14800 16400 Karmegam and Daniel, 2009
Leaf litter of Indian mast tree +
cow dung Perionyx ceylanensis 36.3 43.0 31.5 31.0 1600 1400 15600 16300 Karmegam and Daniel, 2009
Municipal solid waste Eisenia fetida 236.7 201.3 - - 1603 1552 1043 1025 Suthar et al., 2014
Municipal solid waste + cow
dung Eisenia fetida 190.0 169.7 - - 1586 1445 891.3 834.0 Suthar et al., 2014
Paper mill sludge + cattle dung Eisenia andrei 108.0 110.0 34.0 31.0 7500 6900 190.0 180.0 Elvira et al., 1998
Pearl millet cobs + cow dung Lampito mauritii 46.0 63.0 27.0 28.8 2600 1900 800.0 1000 Karmegam and Daniel, 2009
Pearl millet cobs + cow dung Perionyx ceylanensis 52.6 63.0 27.4 28.8 2200 2000 900.0 1000 Karmegam and Daniel, 2009
Poultry droppings + cow dung Eisenia fetida 291.0 179.0 74.1 50.5 1398 1138 - - Yadav and Garg, 2011
Press mud + cow dung Perionyx ceylanensis 44.7 30.6 15.7 8.00 214.5 162.6 17.2 10.2 Prakash and Karmegam, 2010
Rice husk Eudrilus eugeniae - - 11.0 5.00 537.0 258.0 27.0 22.0 Shak et al., 2014
Rice husk + cow dung Eudrilus eugeniae - - 39.0 29.0 3980 2896 401.0 350.0 Shak et al., 2014
Rice straw Eudrilus eugeniae - - 10.0 6.00 2210 1372 280.0 175.0 Shak et al., 2014
28
Substrates Earthworm species Zn mg kg
-1 Cu mg kg-1 Fe mg kg
-1 Mn mg kg-1
References Cast feed Cast feed Cast feed Cast feed
Rice straw + cow dung Eudrilus eugeniae - - 45.0 33 4170 3566 499.0 450.0 Shak et al., 2014
Sewage Metaphire posthuma +
Lampito mauritii 518.4 542.0 35.4 42.6 529.0 563.0 397.4 477.2 Suthar et al., 2008
Sewage sludge Eisenia fetida 289.4 325.6 41.3 48.1 320.8 369.3 - - Suthar, 2010a
Sewage sludge + sugarcane trash Eisenia fetida 96.9 129.5 11.8 18.8 127.7 147.5 - - Suthar, 2010a
Sugar mill filter + horse dung Eisenia fetida 1554 1247 691.0 449 24716 22281 2569 1894 Sangwan et al., 2008a
Water hyacinth Eisenia fetida 243.8 184.2 46.3 45.9 17.1 10.3 585.0 430.1 Singh and Kalamdhad, 2013
Water hyacinth + cattle dung +
sawdust Eisenia fetida 218.1 186.3 46.8 35.6 6.60 6.90 600.0 476.0 Singh and Kalamdhad, 2013
Water hyacinth + cow dung Eudrilus eugeniae 268.0 210.0 96.9 52.1 79.1 71.0 162.3 126.1 Kumar et al., 2015
Winery industry waste Eisenia andrei 62.0 22.0 30.0 22.0 2497 623.0 53.0 8.00 Nogales et al., 2005
29
Suthar (2008a) reported the Zn, Fe, Mn and Cu in
distillery sludge amended with cow dung was
reduced to the extent of 39, 30, 38 and 42%,
respectively after processed by Eisenia fetida. Garg
and Kaushik (2005) and Gupta et al. (2005) also
reported a lower metal contents in the vermicast
compared to the initial feed of solid textile mill
sludge amended with poultry droppings and fly ash
amended with cow dung. Singh and Kalamdhad
(2013) have reported reduction in both total and
extractable form of Cu, Mn, and Fe in
vermicomposted water hyacinth with Eisenia
fetida. The reduction in these metals content was
also reported by other researchers (Jain et al.,
2004). The micronutrient availability in castings
probably determined by the properties of feed they
ingest, might be the reason for observed differential
response of earthworms reported in the previous
art. The reduction of metal nutrients may be
attributed to the tendency of earthworms to
accumulates metals in their tissues during the gut
passage (Hartenstein and Hartenstein, 1981; Graff,
1982; Gupta et al., 2005; Garg and Kaushik, 2005).
3. Effect of ageing on the properties of vermicast
In general, vermicast is rich in available form
of nutrients, beneficial microbial communities,
growth regulators such as auxins, gibberellins,
cytokinins in addition to enhanced physical
properties which can increase the nutrient and water
storage capacity, infiltration and aeration and
resistance to compaction and erosion in soil they
exist (Edwards, 2004). These beneficial properties
possess remarkable plant growth-promoting
potential on wide range of plants (Edwards, 2004;
Gajalakshmi and Abbasi, 2004). However, these
physical, chemical and biological attributes of the
vermicast are not stable in nature, and they change
with time (Hindell et al., 1997a,b; Decaëns et al.,
1999b; Parthasarathi and Ranganathan, 1999, 2000;
Decaëns, 2000; Tiunov and Scheu, 2000a,b;
Scullion et al., 2003; Aira et al., 2005, 2010;
Mariani et al., 2007; Kawaguchi et al., 2011).
Studies on vermicast from either soil or
blends of soil and phytomass have shown that fresh
castings are highly dispersible, which become more
stable during ageing (Marinissen and Dexter, 1990;
Hindell et al., 1997a,b; Piekarz and Lipiec, 2001).
Fresh castings produced by Lumbricus terrestris
and Lumbricus rubellus were up to 1.4 times more
dispersive than soil aggregates (Shipitalo and Protz,
1988), and those of Aporrectodea rosea,
Aporrectodea caliginosa and Aporrectodea
trapezoids were 19 times more dispersive than the
surrounding soil (Hindell et al., 1994). High level
of soluble carbohydrate in the castings was
connected to an increase in dispersion, which is
reported to be disintegrated and bonded to mineral
particles during drying and ageing (Hindell et al.,
1997a,b). Studies on endogeic species,
Aporrectodea caliginosa have shown that fungal
growth, drying and wetting cycle increases the
stability of castings during the ageing (Marinissen
and Dexter, 1990; Piekarz and Lipiec, 2001).
Shipitalo and Protz (1989) have found that ageing
and drying facilitates the bonding of plant and
microbial polysaccharides and other organic
compounds with clay (clay-polyvalent cation-
organic matter), thereby increases the stability of
vermicast. Close association of clay domains with
organic materials reduces the organic matter
decomposition they contain and adding to bond
longevity (Shipitalo and Protz, 1989; Guggenberger
et al., 1996). However, studies on epi-endogeic
earthworm, Lumbricus rubellus revealed that
microbial activity and presence of polysaccharide
did not have any influence on stability of castings
(Marinissen et al., 1996).
Impact of ageing on the bulk density of
casting was significantly varying with castings of
different earthworm species and the feed they
ingest. Decaëns (2000) reported that bulk density of
castings produced by Martiodrilus carimaguensis
was higher or equivalent to that of the surrounding
soil which rely on the organic matter it contains.
The bulk density of casts did not alter in the first
week after deposition, but then decreased
30
progressively to the extent of 29%. Decaëns (2000)
also found that regular shape and thin cortex at their
periphery of castings had largely disappeared
during the ageing. Old castings also contains cracks
and void space which were formed by degradation
of plant debris present in fresh castings and
ingestion of castings by other invertebrates.
Fresh vermicast are reported to contain
higher content of organic carbon than its
surrounding soil (Lee, 1985; Shipitalo and Protz,
1988; Daniel and Anderson, 1992; Barois et al.,
1993; Zhang and Schrader, 1993; Buck et al., 1999;
Bossuyt et al., 2005), whereas castings from purely
phytomass or manure have showed reduction in
carbon content than the feed they ingest (Aira and
Domínguez, 2009). Contradiction in the results is
due to the preferential ingestion of high organic
matter by earthworm in field, and however, C
content of castings compare with only the bulk soil.
During ageing, the organic carbon present in the
castings mineralized by microbial activity, leading
to reduction in the carbon pool in the vermicast
(McInerney and Bolger, 2000a). However, in the
studies on castings deposited in nature, a
continuous increase in C was observed during their
ageing (Jiménez and Decaëns, 2004). The reason
may be fixation of atmospheric CO2 by algae or
nitrification bacteria (autotrophic microorganisms);
colonization of casts by cast-dwelling macro-
invertebrates and accumulation of organic material
and/or the production of carbon-enriched feces by
earthworms fed on organic-rich food substrates
(Jiménez and Decaëns, 2004). The organic matter
in the ageing vermicast can be protected over
longer periods of time from further decomposition
and might then become available for the microflora
once these are broken down into small fragments
(Blanchart et al., 1997; Decaëns, 2000; Bossuyt et
al., 2005).
The vermicast are widely reported to contain
elevated amount of mineral nitrogen, especially for
the ammonium content relative to surrounding soil
(Parkin and Berry, 1994; Decaëns et al., 1999a;
Whalen et al., 2000; Aira et al., 2003; Bityutskii et
al., 2012; Clause et al., 2013). During ageing, the
casts were characterized by a high rate of
nitrification, which reduced the ammonium content,
and simultaneously increase nitrates. Kharin and
Kurakov (2009) have reported that the castings of
endogeic earthworm, Aporrectodea caliginosa
showed two to threefold increase in nitrate content
during the 12 days of ageing. Further ageing,
reduced the denitrification activity, and became
closer to their surrounding soil. The nitrogen
present in microbial biomass also greatly affect by
the ageing, thus in 45 days old castings of
Aporrectodea caliginosa it reduced to the extent of
33% (Aira et al., 2005). The activities of enzymes
such as dehydrogenase, β-glucosidase, cellulase,
invertase, amylase, protease, phosphatase of
castings were also reported to reduce during the
ageing (Aira et al., 2005; Kharin and Kurakov,
2009). The reduced enzyme activities in aged casts
were probably due to: (i) the reduced nutrient and
moisture contents, (ii) a decline or inactivation of
the microbial population, (iii) low stability of aged
earthworm casts, and (iv) degradation or
inactivation of enzymes during the ageing (Aira et
al., 2005).
Microbial diversity and its activity in castings
were strongly affected by age. Fresh castings are
reported to contain higher microbial biomass than
the feed they ingest (Parthasarathi and
Ranganathan, 1999; Tiunov and Scheu, 2000a,b).
Ageing have been showed predictable microbial
successions with castings of different species.
During ageing, the bacterial activity reduced due to
shifting from bacterial to fungal dominance within
the population (Scullion et al., 2003; Piekarz and
Lipiec, 2001; Aira et al., 2005). In the castings of
Lumbricus terrestris and Lumbricus rubellus, the
bacterial population largely increased in the first
four days and then there was rapid decline (Scullion
et al., 2003). Similarly, the castings of
Aporrectodea caliginosa also showed increase in
bacterial population in the initial days followed by
fungal dominance over the bacterial population
31
during the first week of ageing. Even though,
further ageing reduced the fungal population, it
remained four times more than that in the soil
during the 12 days ageing (Kharin and Kurakov,
2009). Castings deposited in the field are found to
be colonized by macro-invertebrates after 4 to 6
weeks of ageing. The density of invertebrates found
inside casts was reported to much lower than those
found in soil. The total count of individuals inside
the casts or in the underlying soil was not changed
during cast ageing. Moreover, only a small number
of specialized (i.e. small and mobile) species are
able to live inside vermicast (Decaëns et al.,
1999b).
4. Effect of vermicast on plant growth and soil
Vermicast contains all the essential macro
and micronutrients for plants growth, regardless of
different substrate used to generating vermicast.
The nutrient present in the vermicast are more
bioavailable (Gajalakshmi and Abbasi, 2008;
Edward et al., 2011), and the nutrients bound in
organic matter are release gradually through
mineralization of the organic matter, thus causing
lesser nutrient loss from the rhizosphere (Chaoui et
al., 2003). Vermicompost also improves the
physical properties of the soil, such as aeration,
water holding capacity and porosity which all have
direct impact on the plant productivity in vermicast
applied soil (Edwards, 2004). Additionally the
beneficial impact of vermicast is also attributed to
biologically active substances such as fulvic acids,
humic acids and phytohormones it contains.
Specifically, cytokinins have been reported in
vermicast (Zhang et al., 2014). It has also been
shown that humic acids derived from the vermicast
induce morphogenetic and biological changes
favorable to plants which are similar to those
produced by indole-3-acetic acid (Muscolo et al.,
1999). These bioactive substances are probably
produced due to the abundance of microbial
communities in the vermicompost, specifically the
actinomycetes and fungal species, which then
releases phytohormones in the soil (Frankenberger
and Arshad, 1995). Enhancement of all these
properties of soil with vermicast supplement are
reported to increase productivity of many plants
(Tomati et al., 1983, 1988, 1990, 1995; Abbasi and
Ramasamy, 1999; Atiyeh et al., 2002; Arancon et
al., 2003, 2004; Gajalakshmi and Abbasi, 2004;
Acevedo and Pire, 2004; Edwards, 2004; Sinha,
2009).
It is also postulated that vermicompost can
suppress a wide range of microbial diseases, insect
pests and plant parasitic nematodes. Availability of
adequate nutrients with vermicast application
enhances the ability of the plants to limit the
penetration, development, and/or reproduction of
invading pathogens (Graham and Webb, 1991),
which contributes to the development of resistance
to pathogens in the plants. Some of the nutrients are
also involved in the production of antimicrobial
compounds such as flavonoids and phenolics that
act against the plant pathogens (Graham and Webb,
1991; Hill et al., 1999). The humic acid content of
the vermicast is also likely to affect biochemical
processes in the plants and bacteria, resulting in
induction of resistance in plants to certain
phytopathogens (Sahni et al., 2008). Vermicast
application also enhances the activity of diverse
beneficial microbes in soil, which may act as
biocontrol agents to suppress plant pathogens
(Gunadi et al., 2002; Arancon et al., 2006). Prior
art shows that the vermicast significantly reduced
sporulation of the pathogen Phytophthora
cryptogea (Orlikowski, 1999), reducing the growth
of pathogenic fungi such as Botrytis cinerea,
Sclerotinia sclerotiorum, Corticium rolfsii,
Rhizoctonia solani and Fusarium oxysporum
(Nakasone et al., 1999), reduced infection of
Fusarium lycopersici (Szczech, 1999) and
Phytophthora nicotianae (Szczech and Smolinska,
2001) in tomato seedlings. Vermicast also reported
to be reducing the infestation of Heteropsylla
cubana (Biradar et al., 1998), Aproaerema
modicella (Ramesh, 2000), and many other insect
pests and mites in various plants.
32
Vermicast effect on density and water related
properties on the soil rely on castings age and
species that produce them. Aged castings are more
stable than its surrounding soil, whereas the fresh
castings are easily dispersible, in water, and thereby
effecting clogging of transmission pore and
impeding the infiltration in soil (Shipitalo and
Protz, 1988; Le Bayon and Binet, 1999). The aged
castings being stable get protected from detachment
by rainfall or surface flow, and as a consequence
increase infiltration. The shape, size and
compaction of castings also have differential
impact on soil properties. Castings produced by
‗compacting‘ species such as Pontoscolex
corethrurus and Millsonia anomala are reported to
impede the infiltration, increase the bulk density
and reduce the porosity of soil (Alegre et al., 1995;
Barros et al., 1996; Young et al., 1998; Chauvel et
al., 1999; Hallaire et al., 2000; Spain et al., 1992;
Derouard et al., 1997; Blanchart et al., 2004). In
contrast, larger size compacting type castings
produced by Amynthas khami are reported to
improve water infiltration by their large packing
voids between castings which are connected with
below-ground galleries (Jouquet et al., 2008);
whereas, the castings of decompacting type are
highly dispersible and impedes the infiltration rate
(Shipitalo and Protz, 1988; Le Bayon and Binet,
1999). The large size castings of compacting
species also create anaerobic condition inside the
castings, which reduces the rate of decomposition
of organic matter it contains (Blanchart et al.,
1993). In the case of smaller castings produced by
the decompacting species, the decomposition rate is
accelerated due to high microbial and enzyme
activity along with favorable physical condition for
the growth of aerobic microorganisms. Similarly, in
the fresh castings, the organic matter has fast
decomposition rate and it is stabilized within
microaggregates formed within the casts during the
ageing (Six et al., 2014). Thus, the casting of
different age of different species also has
differential impact on the soil carbon stock.
5. Conclusions
Vermicast produced by different earthworm
species has different physical, chemical, biological
characteristics, which rely on the habitat and
feeding behavior of earthworms that produce them.
Thus, the impact of castings of different earthworm
species on the soil and plant also vary significantly.
In general, castings of all the species have higher
plant available nutrient than their surrounding soil
or the feed they ingest. The presence of growth
regulators, beneficial microbes, and enhanced
enzymes activity associated with nutrient
mineralization are also reported in castings of many
species. Castings also improve the physical
properties of soil by reducing bulk density and
erosion, enhancing porosity, infiltration, water
holding capacity and so on. However, most of the
previous studies on vermicast in relation to soil
properties such infiltration, compaction, erosion,
soil carbon stock, structural changes and other
related aspects have been studied with anecic and
endogeic species. While, the castings of epigeics
has been extensively studied in relation to plant
growth. But not much focus has been paid to
understand the impact of castings of anecic and
endogeic species on seed germination, plant growth
or disease suppression, or vis-à-vis.
Moreover, the physical, chemical and
biological attributes of the vermicast are not stable
in nature, and they change with cast ageing.
Previous studies on the fate of vermicast over time,
have mainly focused on the stability of castings
deposited by anecic and endogeic earthworm
species and very few studies have been done on
epigeics. The castings used in these studies were
also produced from non-specific substrates in
nature or from blends of soil and phytomass.
Therefore, the castings may contain considerable
amount of soil particles, which is known to increase
the stability of these biogenic structure, slow down
the decomposition of organic matter, and also either
33
stabilize or bring about proteolysis of the
extracellular enzymes it contains. Hence, the
findings of these studies cannot be used as a model
to study the dynamics of the properties of castings
when it is produced by anthropogenically
controlled vermicomposting systems, which are
purely derived from the organic matter. It is
therefore necessary to conduct more studies on
these aspects to improve our understanding on the
interaction of these biogenic structures with agro-
ecosystems, so that the beneficial features of
vermicast can be utilized maximally.
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dispersion of disturbed soils. J. Hydrol. 260,
194-205.
52
INGESTION OF SAND AND SOIL BY PHYTOPHAGOUS
EARTHWORM EUDRILUS EUGENIAE: A FINDING OF
RELEVANCE TO EARTHWORM ECOLOGY AS WELL AS
VERMITECHNOLOGY
Chapter
3
53
A paper based on this chapter has been published in
Archives of Agronomy and Soil Science, 60 (12), 1795 – 1804, 2014.
CChhaapptteerr 33
Ingestion of sand and soil by phytophagous earthworm
Eudrilus eugeniae: a finding of relevance to earthworm
ecology as well as vermitechnology
Abstract
Epigeic earthworms are phytophagous in habit and are believed to prefer organic matter, principally
phytomass, in various degrees of decay. To check whether epigeics will ingest soil/sand even when there is
luxury availability of phytomass, the present work was carried out. It was seen that not only soil but even
sand particles were ingested by epigeic (Eudrilus eugeniae) tested by us, even as there was availability of
phytomass and cow manure in abundance. But the consumption of sand and soil decreased as the days
progressed. Moreover, the ingestion of sand and soil with phytomass and cow dung did not show any
significant influence on growth and survival of earthworms. It did increase the vermicast mass due to the
presence of soil/sand as was confirmed by microphotography using polarizing microscope. The findings have
important implications in the design of vermireactors for maximizing their efficiency.
1. Introduction
Epigeics are humus feeding worms which
dwell at or very near the surface of the soil horizon.
Due to their preference for phytomass as food,
epigeics are also called phytophagous in contrast to
the anecics which are termed geophytophagous due
to their tendency to ingest soil together with
organic matter, and endogeics which are called
geophagous due to their preference for soil. It is
generally believed that periodic ingestion of
sand/soil is essential for even epigeics to keep their
gizzard muscles toned up. As gizzards play a
pivotal role in comminuting the material that the
earthworm ingest, it has been believed that hard-to-
break particulates as are contained in sand/soil
should be always a part of earthworm diet if they
have to remain healthy (Marhan and Scheu, 2005).
Although this belief is well-entrenched, there is no
existing report which establishes its veracity. This
is despite the fact that the role of epigeics in the
degradation of various types of leaf litter and other
organic wastes has been explored in detail (Lim et
al., 2012, 2014; Shak et al., 2014). Information is
also available on the attempts to use fly ash (Saxena
et al., 1998; Gupta et al., 2005), rock phosphate
(Wang et al., 2013), and sludges of textile mills
(Garg and Kaushik, 2005), tanneries (Hemalatha
and Meenabal, 2005), petrochemical industries
(Banu et al., 2005), pharmaceutical industries
(Majumdar et al., 2006), and distilleries (Suthar and
Singh, 2008) as epigeic feed after amending it with
animal manure or sewage sludge. But not much
attention has been paid towards determining
whether ingestion of soil/sand is essential for long-
term survival and fecundity of epigeics.
Eudrilus eugeniae is known to be a voracious
feeder of animal manure, especially cow dung
(Fernández-Gómez et al., 2013), but it’s acceptance
for phytomass is not as well- established. Indeed
except a few studies by Gajalakshmi, S.A. Abbasi
and co-workers (Gajalakshmi et al., 2002; Makhija
et al., 2011), no one has reported direct
54
vermicomposting of phytomass – without pre-
composting or subsequent blending with animal
manure – by E. eugeniae. On the other hand, by
employing the concept of high-rate
vermicomposting and associated technology
developed by Abbasi and co-workers (Gajalakshmi
et al., 2002, 2005; Abbasi et al., 2009, 2011;
Tauseef et al., 2013), several types of phytomass
have been recently vermicomposted using
E.eugeniae and other species in reactors which had
no sand/soil or any other bedding, and which were
fed unprocessed phytomass in the form of leaf
litter, weeds, vegetable waste etc. Also the reactors
could be run in pulse-fed mode for as long as one
wished, with the epigeic species placed in them not
only surviving but gaining in zoomass and
exhibiting good fecundity. Hence, in the context of
vermireactors design for attaining maximum
vermiconversion efficiency it has become important
to answer the question: Is presence of sand/soil
really essential? There is also a need to establish
whether E. eugeniae may ingest soil/sand if forced
to feed upon phytomass but may not do so when
fed with animal manure. Hence, we chose cow
manure, leaves of neem (Azadirachta indica) and of
ipomoea (Ipomoea carnea) as substrates for the
present study. Neem and ipomoea were chosen
because both are likely to be inimical to, rather than
preferred by, E. eugeniae as neem contains several
chemicals known to repel or kill invertebrates while
ipomoea contains toxic alkaloids. We also studied
the influence of different earthworm densities (25,
15 and 5 animals per kg of substrate) on feed
consumption, as crowding may induce a tendency
to forage beyond the phytomass layer and into the
sand/soil bedding. Of the epigeic species studied
most extensively vis-a-vis vermicomposting so far
– Eisenia fetida (Savigny), E. eugeniae (Kinberg)
and Perionyx excavatus (Perrier) (Sim and Wu,
2010) – E. eugeniae was chosen by us as it is the
most voracious of feeders among the three
(Gajalakshmi et al., 2002, 2005; Edwards, 2004;
Lim et al., 2011). In addition, epigeics may have
advantage over burrowing earthworms in cutaneous
absorption and dietary intake due to their surface-
dwelling mode of life (Suthar, 2014).
2. Materials and methods
Ipomoea and neem leaves were collected
from in and around the Pondicherry University
campus. The leaves were washed with water to
remove adhering material, then soaked for 48 hours
and lightly squeezed to soften them. Rectangular,
41 liter wooden boxes (30 cm high with surface
area of 35 x 39 cm) were used as vermireactors.
They were lined up with thick transparent plastic
sheets to prevent water loss, as also to prevent
earthworms from escaping or predators from
coming in.
Five sets of vermireactors were employed.
The first set consisted of reactors which were filled
with a layer of coarse sand to a height of 3 cm
followed by a 5 cm high layer of soil. Over it 1kg
dry weight equivalent of cow dung was placed
which has raised the reactor content further by
about 7 cm. Other reactors in this set consisted of
ipomoea or neem instead of cow dung. The heights
of the substrates in these reactors were 12 and 7.5
cm, respectively. The second set of reactors was
identical to the first set except that there were
without sand/soil. The third set comprised of the
control reactors which had only soil and sand
bedding but no substrate. Bunches of 25 adult
earthworms, weighing 0.9 - 1.5 g and length of 9 -
12 cm, randomly picked from their cow dung –
based culture, were introduced in each of the
reactors. The fourth and fifth sets of reactors were
identical with the first set except that the number of
earthworms incorporated therein was 15 and 5
separately.
All the reactors were operated at identical
ambient conditions with temperature of 29 ± 4°C.
Moisture was maintained by periodic sprinkling of
adequate quantities of water. The reactors were
operated continuously for 8 runs of 15 days each.
At the end of each run, vermicast in all the reactors
55
Table 1. Major and trace elements composition of plant leaves, cow dung, soil and sand used in this study
Parameters Neem leaf Ipomoea leaf Cow dung Soil Sand
Dry matter % 34.0±0.4 21.9±0.2 26.5±0.3 98.6±0.0 98.3±0.2
Organic carbon mg g-1
616.1±5.9 656.9±4.4 359.9±8.0 4.30±0.07 0.39±0.15
Nitrogen mg g-1
32.7±1.5 39.6±2.3 25.4±0.5 0.34±0.10 0.11±0.06
Phosphorus mg g-1
0.20±0.00 0.26±0.01 0.62±0.00 0.25±0.03 0.16±0.01
Potassium mg g-1
16.0±0.1 39.2±0.3 7.38±0.42 5.49±0.15 15.0±0.4
Sulphur mg g-1
6.58±0.05 6.89±0.34 7.78±0.42 0.07±0.06 0.16±0.05
Calcium mg g-1
16.5±0.2 11.2±0.3 4.91±0.23 3.51±0.06 12.8±0.7
Sodium mg g-1
0.56±0.03 0.73±0.05 3.94±0.09 2.45±0.02 11.4±0.6
Magnesium mg g-1
6.58±0.14 2.48±0.04 4.37±0.18 1.04±0.03 3.47±0.36
Copper mg kg-1
7.89±1.69 21.1±1. 6 25.1±4.4 – 33.3±28.9
Iron mg g-1
0.22±0.01 0.36±0.00 4.75±0.42 63.0±0.7 30.7±24.0
Manganese mg kg-1
44.8±1.6 85.2±4.8 274.0±10.9 0.03±0.02 200.0±173.5
Zinc mg kg-1
29.1±0.7 34.1±3.9 69.0±2.6 0.01±0.00 10.0±17.3
was harvested and the reactors were restarted with
the leftover substrate. The castings were carefully
separated from other particles by soft painting
brush and quantified. While disbanding the
reactors, the adult earthworms with which the
experiment was started, were washed, blotted dry
and weighed to record their zoomass before they
were put back in the respective reactors. Dead
earthworms, if any present, were replaced, while
starting the next run.
Total organic carbon content in the soil, sand
and plant material was measured by modified
dichromate redox method (Heanes, 1984). Total
nitrogen content was determined by modified
Kjeldahl method (Kandeler, 1993) using Kel Plus™
semi-automated digester and distillation units
(Pelican Equipments, Chennai, India). Total
phosphorus content of the plant material was
determined by ammonium molybdate –
hydroquinone method (AOAC, 2012). Potassium,
calcium and sodium were determined using an
Elico CL378 model flame photometer (Elico Ltd,
Hyderabad, India) after dry ashing the plant
material at 500°C for 4 hours and dissolution of the
ash in hydrochloric acid (Kalra, 1998). Magnesium,
boron, copper, iron, manganese, zinc and
molybdenum concentration in plant samples were
determined with thermo electron IRIS intrepid II
XSP DUO model inductively coupled plasma
atomic emission spectroscopy (ICP-AES, Thermo
Fisher, Waltham, MA, USA) as per procedure
given in AOAC (2012). The soil and sand samples
were analyzed for their mineral content by
Bruker™ S4-Pioneer model wavelength dispersive
X-ray fluorescence spectrophotometer (WD-XRF,
Bruker, Billerica, MA, USA). The samples were
ground to 100 µm particle size using ball mill. The
processing of emission spectra for identification of
elements and their peak area measurement was
carried out using SPECTRAplus® software, version
1.6 (Pioneer Hill Software, Poulsbo, WA USA).
The results are summarized in Table 1.
The presence of sand, soil and organic matter
in the vermicast was quantified by gravimetry.
Samples oven dried at 105 °C to a constant weight
were put in distilled water, and crushed to release
the soil/sand. The content was filtered through a
Whatman No. 42 filter paper. The residue over the
filter paper was kept in a muffle furnace at 550°C
for 4 hours to remove organic matter present in the
castings (John, 2004). To observe the internal
structure of casts, 1mm thin sections of the castings
were made, after impregnating them in araldite –
xylene mixture, with the help of Buehler
56
PetroThin® thin sectioning system (Buehler, Lake
Bluff, IL, USA). They were studied using a
polarizing microscope at low magnification (4x)
(FitzPatrick, 1993). The surface area of casting and
sand grains in the sections were measured with the
Leica® software, version 3.8.0 (Leica
Microsystems, Heerbrugg, Switzerland).
Differences in the significance of vermicast
production, earthworm mortality, zoomass gain and
sand and soil content in the vermicast between
different treatments were tested by repeated
analysis of variance. The cast size and grain
covered area in the vermicast sections were
analyzed with two-way ANOVA at the 99.9%
confidential level. SPSS 16 package (Softonic,
Barcelona, Spain) was used for all statistical
analysis.
3. Results and discussion
3.1. Vermicast output
There was significant difference (p<0.01) in
vermicast production between reactors with
different substrates, worm density and presence and
absence of sand/soil bedding (Figure1 and Table 2)
indicating the influence of these factors on
vermicast production. In general, the chemical
nature of the organic waste influences the
palatability of earthworms directly or indirectly,
which consequently affects the earthworms’
efficiency in processing a substrate. In the present
study, vermireactors with the presence of sand/soil
bedding, produced maximum amount of vermicast
in conjunction with cow dung, followed by neem
and ipomoea. This is consistent with the belief that
earthworms are attracted by most kinds of animal
dung (Lowe and Butt, 2005; Marhan and Scheu,
2005), due to partially decomposed nature and high
nitrogenous organic matter content of animal dung.
The quantity of cast generated with cow dung as
feed and with sand and soil bedding was 643±100,
701±70 and 1392±10 mg cast worm
-1 d
-1 with 25,
15 and 5 worms per kg dry weight of substrate,
respectively. The lowest cast production was seen
when the ipomoea was offered as a feed: 251±10,
308±70 and 423±80 mg cast worm-1
d-1
castings
with 25, 15 and 5 worms, respectively. Ipomoea is
reported to consist a variety of toxic alkaloids such
B1, B2, B3, C1 and N-methyi-trans-4-hydroxy-L-
proline (Asano et al., 1995; Molyneux et al., 1995;
Cholich et al., 2013). It is possible that the presence
Figure 1. Vermicast output, g worm -1
day-1
(mean ± SD) recorded in reactors with different treatments.
(‘w’ represent the worm density per kg substrate).
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
5w 15w 25w 5w 15w 25w 5w 15w 25w 25w 25w 25w 25w
Neem Cow dung Ipomoea Neem Cow dung
Ipomoea
With Bedding Without Bedding Bedding alone
Ve
rmic
ast
ou
tpu
t, g
w -1
d-1
Treatments
57
Table 2. Repeated analysis of variants and ANOVA table of F-values and the effects of substrate, bedding
and worm density on vermicast output, average zoomass changes, mortality, sand and soil entrapped in
castings, castings surface area and sand grains covered area of castings.
Repeated analysis of variants ANOVA
df Vermicast
output
Average
zoomass
changes
Mortality
Sand and
soil
entrapped
Casting
surface
area
Sand grains
covered
area
Substrate 2 63.18***
152.0***
16.27**
123.47***
105.31***
102.44***
Bedding 1 36.87**
7.58 n.s
215.4***
– 12.75***
–
Worm density 2 60.23***
9.76**
1.01n.s
1.27n.s
4.33* 3.50
*
Substrate x bedding 1 2.80 n.s
0.201 n.s
30.99***
– 37.41***
–
Substrate x worm density 4 3.12 n.s
6.38* 2.019
n.s 2.03
n.s 3.243
* 3.41
**
Bedding x substrate x
worm density 6 – – – – 17.86
*** –
*p<0.05,
**p<0.01,
***p<0.001,
n.s - not significant.
of these chemicals led to less intense feeding and
lower cast production in comparison to the other
substrates.
In the case of neem-based reactors, the
amount of cast production was higher than in
ipomoea but lower than in cow dung. In these
reactors 496±40, 622±40 and 1037±80 mg
cast
worm-1
d-1
were produced with 25, 15 and 5 worms,
respectively. Even though neem is known to have
nematicidal and insecticidal properties (Dai et al.,
2001; Nathan et al., 2005) it does not seem to have
a deleterious effect on earthworms. Earlier
Gajalakshmi and Abbasi (2004) and
Selvamuthukumaran and Neelanarayanan (2012)
have reported that earthworms fed upon the neem
leaf compost even more voraciously than they did
on the compost of mango leaf and ashoka tree leaf.
However, the presence of polyphenolic compounds
might have affected the earthworm in fresh
substrate. The antagonism between earthworms and
polyphenols/lignin content of their feed has been
reported in studies by Satchell and Low (1967),
Hendriksen (1990), Tian et al. (2000) and Ganesh
et al. (2009). These studies showed a negative
correlation between the palatability of leaf litter and
its total polyphenol content in both natural and
planted fallows.
The pattern of vermicast output from
different treatments is illustrated in Figure 1. The
results of cast generated per worm per day with
different earthworm densities showed decreasing
trend with increasing worm density. The maximum
vermicast generated was 1392±10, 1037±80 and
423±80 mg cast worm
-1 d
-1 in cow-manure, neem-
and ipomoea-based reactors, respectively, run with
5 earthworms. Minimum vermicast productions of
643±100, 496±40 and 251±10 mg cast worm
-1 d
-1
were observed in cow manure, neem and ipomoea,
respectively, in reactors with 25 earthworms. The
results showed remarkable differences between the
masses of vermicast generation in reactors which
consisted sand/soil and ones which did not.
Reactors without sand/soil and with 25 worms
yielded 316±20 and 310±40 mg worm
-1 d
-1 cast
with cow manure and neem, respectively. A
significant amount of sand and soil entrapped in the
castings may be the reason for the higher mass of
vermicast obtained from reactors with sand/soil
bedding compared to the ones without this bedding.
However, there was no significant difference in net
58
output of vermicast from reactors with and without
sand/soil. Very few castings were seen in ipomoea-
fed reactors without bedding, due to the 100%
mortality in all the three runs. In the case of control,
very few castings were observed. The casts were
less stable and easily disintegrated while
moistening the reactors. Hence, the castings
generated in the control reactors could not be
studied further.
3.2. Mortality
There was mortality of earthworms in the
first run in most of the reactors. The type of
substrate and the presence of sand/soil had a
significant effect on the survival of the earthworms
(p<0.01) (Table 2). The difference was more
pronounced in ipomoea-based reactors. Ipomoea
reactors with sand/soil showed 100% and ~40%
mortality in the first and second runs, respectively,
but had no mortality from then onwards. In contrast
in ipomoea reactors without sand/soil, 100%
mortality was observed consistently from the
beginning to third run. Hence, these reactors were
not operated further. In neem reactors with bedding,
a maximum of 84% mortality was seen in one of
the reactors with 25 earthworms and in the
remaining reactors there was no significant
mortality in the first run. In the case of reactors
without sand/soil, a maximum of 52% mortality
was recorded in the first run. However in reactors
with and without sand/soil, there was no mortality
from the second run onwards. In the case of cow
dung-fed reactors, there was no mortality in any of
the runs. The reactors with soil only as bedding and
without substrate did not show any mortality up to
third run, but from fourth run, the mortality rate
increased in these reactors. The reason may be that
in these reactors the earthworms, which are
phytophagous, were forced to feed exclusively on
sand/soil and could not survive on it for long.
3.3. Average zoomass change per animal
There was a significant influence (p<0.01) of
different substrates and worm densities on
earthworm zoomass, but there was no difference in
zoomass between reactors with and without
sand/soil (Table 2). In the first run, there was a
decrease in zoomass in most of the reactors with
neem and ipomoea, but from the second run
onwards there was a gain in zoomass in all the
reactors. With cow dung as feed, the increase in
zoomass was recorded in all the reactors from the
beginning to the eighth run. The results also show
that there was no significant difference in zoomass
change between reactors with 5 and 15 worms;
however, further increase in worm density to 25
showed a decline in zoomass gain in comparison to
the lower worm densities.
Even though the earthworms fed on ipomoea
leaves in reactors which also had soil and sand, and
produced a considerable amount of vermicast in all
the runs, there was a steady decrease in zoomass in
all ipomoea-based reactors. It is likely that the
ipomoea employed in these reactors had
accumulated toxic inorganics or heavy metals as it
was picked up from near high ways and chemical
industries. This might have caused the adverse
impacts. But as expected, cow manure was
preferred by earthworm over neem and ipomoea.
3.4. Surface area of vermicast
The surface area of the castings varied
significantly between feed to feed and was also
influenced by the presence of sand/soil (p<0.001)
(Table 2). The castings produced from reactors fed
with ipomoea, sand and soil had a significantly
higher surface area in comparison to others of that
set, and ranged from 16730.3±2384.2 to
17202.735±2069 µm2. Cow manure and neem fed
earthworms produced castings with surface areas
59
Figure 2. Percentage of sand and soil particle entrapped in the castings measured by gravimetric method
(mean ± SD) and percentage of sand particle covered area in the castings measured by thin-sectioning
method (mean ± SD). (‘w’ represent the worm density per kg substrate).
ranging 10297.8±171.8 to 13016±649.57 µm2 and
10112.5±2519.7 to 14125.5±1053 µm2,
respectively. The casts produced from the reactors
without sand and soil was significantly smaller in
size: 8651.2±1299.3µm2 and 7991.6±1537.6 µm
2
for cow dung and neem respectively. The size of
the castings positively correlated with the amount
of sand/soil ingested by the earthworms. The
difference in worm density did not show any
variable effect on cast size.
3.5. Assimilation of sand and soil particles in
castings
The percentage of sand and soil assimilation
in castings and the percentage of sand covered area
in vermicast sections are presented in Figure 2. A
clear difference can be seen between the amounts
of sand assimilated in castings of different feeds (p
< 0.001) (Table 2). Castings from ipomoea had the
largest sand covered area (Figure 3) of 17.7±1.17%
to 26.2±3.71%, followed by cow dung and neem
ranged from 8.93±3.33 to 13.8±2.06% and
6.57±0.27% to 10.1±2.29%, respectively. The
extent of sand assimilation in castings of cow-
manure and neem-fed earthworms differed slightly
but the difference was not statistically significant.
A similar trend was revealed by gravimetric
analysis of sand and soil entrapped in the vermicast.
The castings from ipomoea-fed reactors had
51.09±3.2%, 43.26±3.3% and 47.67±3.3% of sand
and soil when earthworm densities were 25, 15 and
5, respectively. The corresponding figures for cow
dung and neem vermicast were 26.79±3.6%,
24.23±0.6%, 22.73±1.6%, and 30.93±3.6%,
28.67±1.4%, 29.05±3.3%, respectively. As for
control reactors which has only soil and sand the
castings could not be harvested intact from the
reactors and hence their shape and size could not be
quantified. Irrespective of the size of the castings,
the area covered in it by sand differed significantly
in different treatments. The highest sand-covered
area was observed in castings from ipomoea:
9473.18 µm2. With cow manure and neem feed it
was 6047.72 and 5392.54 µm2 respectively; the
difference not being statistically significant. In the
present studies the earthworms were given access to
0
10
20
30
40
50
60
5w 15w 25w 5w 15w 25w 5w 15w 25w
Neem Cowdung Ipomoea
San
d/s
oil
en
trap
pe
d in
cas
tin
gs (
%)
Treatments
Sand and soil entrapped (%) Sand grains covered area (%)
60
Figure 3. The thin sectioned vermicast from neem, cow dung and ipomoea based reactors with soil + sand
bedding illustrated in a, b and c. The light – colored portions inside the castings are mineral soil particles.
The castings from the reactor without soil/sand showing in d.
Figure 4. Regression analysis between the sand and soil entrapped in the castings (%) and (a) the average
zoomass changes and (b) mortality rate.
y = -100.4x + 37.204 R² = 0.9201
0
10
20
30
40
50
60
-0.15 -0.1 -0.05 0 0.05 0.1 0.15 0.2
San
d a
nd
so
il e
ntr
app
ed
in c
asti
ngs
(%
)
Average zoomass changes worm-1 run-1
y = 1.3506x + 23.029 R² = 0.8916
0
10
20
30
40
50
60
0 5 10 15 20
San
d a
nd
so
il e
ntr
app
ed
in c
asti
ngs
(%
)
Average mortality (%)
61
both sand and soil and separate studies on sand or
soil were not carried out. But it can be surmised
that broad trends vis-a-vis pattern of ingestion
would be no different than seen in the present
study. When anecics ingest sand along with plant
debris and grind their feed, the attrition caused by
sand results in very fine fragmentation of organic
matter, enhancing its surface area and making it
much more amenable to microbial action than was
otherwise possible (Schulmann and Tiunov, 1999).
Had a similar process been occurring in the
presence study, the ingestion of sand and soil along
with neem and cow dung would have facilitated
digestion of the phytomass. But such was not the
case and there was no significant difference in
zoomass gain between reactors with and without
soil/sand bedding. The increase in sand
consumption rate in ipomoea-fed reactors may be to
fulfill their energetic requirements and avoid
consumption of ipomoea leaves. This tendency has
been seen in several other species (Curry and
Schmidt, 2007). A regression analysis between the
amount of sand and soil consumed and average
zoomass change in earthworms and mortality rate
substantiates these findings (Figure 4).
4. Conclusions
Controlled experiments were carried out to
see whether the epigeic earthworm – E. eugeniae
would ingest sand and soil even when phytomass
was available in abundance. It was seen that even
though initially E. eugeniae did ingest sand and soil
despite the luxury availability of phytomass, this
tendency was reduced as the time passed indicating
adaptive response to the phytomass feed.
These findings are of significance in
vermireactor’s design and optimization because
they indicate that sand-soil-gravel bedding as used
in conventional vermireactors is not really
necessary to ensure the survival, growth and
fecundity of the epigeics – used in the
vermicomposting. The findings reinforced the
validity of the high-rate vermicomposting concept
and associated nuances of vermireactors design
introduced earlier by the authors.
References
Abbasi, T., Gajalakshmi, S., Abbasi, S.A., 2009.
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64
FEEDING BEHAVIOUR OF PHYTOPHAGOUS
EARTHWORM EUDRILUS EUGENIAE IN HIGH-
SUBSTRATE COLUMN VERMIREACTORS
Chapter
4
65
A paper based on this chapter has been
submitted for publication.
CChhaapptteerr 44
Feeding behaviour of phytophagous earthworm Eudrilus
eugeniae in high-substrate column vermireactors
Abstract
In chapter 3 study made to check whether the epigeic earthworm, Eudrilus eugeniae will ingest soil/sand
even when there is availability of phytomass in plenty is reported. It was seen that the worms, ingested not
only soil even sand particles along feed. Moreover, the soil/sand ingestion rate was significantly influenced
by the substrate type – cow manure, leaves of neem and of ipomoea – tested by us. Since, the findings have
important implications in the design of vermireactors for maximizing their efficiency; the influence of
vemireactors’ constituent on the epigeic’s feeding behavior is of the essence, which needs to be resolved. In
this chapter, study on the influence of (a) feed height in the vermireactors (b) the mode of supply of feed, and
(c) the degree of decomposition of the feed, on the preference towards sand and soil by the epigeic
earthworm, E. eugeniae is reported.
1. Introduction
Earthworms are one of the most important
component of soil biota in terms of soil formation
and maintenance of soil structure and fertility
(Edwards, 2004). The function of earthworms on
soil ecosystem differs considerably with different
earthworm species that are linked to their feeding
strategy (Uchida et al., 2004). The earthworms
mainly feed on organic matter together with
microorganisms, micro and meso fauna, nematodes
and their dead remains (Curry and Schmidt, 2007).
However, anecics and endogeics species which
often live in subsoil, consume an appreciable
amount of mineral soil, even in the presence of
phytomass and animal manure (Hendriksen, 1991;
Schulmann and Tiunov,1999), possibly to keep
their gizzard muscles toned up (Marhan and Scheu,
2005). The epigeics are considered to ingest only
organic matter in various degrees of decay.
As described in chapter 3, study was
conducted to check whether the epigeic
earthworm, Eudrilus eugeniae will ingest soil/sand
even when there is luxury availability of
phytomass. It was seen that initially the E. eugeniae
did ingest sand and soil like anecic and endogeic
species, despite the availability of phytomass in
plenty. The amount of soil/sand ingestion
significantly varied with different feed – leaves of
neem and of ipomoea, and cow manure. As the
days progressed, the consumption of sand and soil
decreased with all the three feed. Influence of
different earthworm densities on soil/sand
consumption was also studied, and it did not show
any variable effect on sand /soil consumption.
Since these studies have important
implications in the design of vermireactors for
maximizing their efficiency, it is necessary to
understand the influence of vemireactors’
constituent on the epigeic’s feeding behavior.
Hence in the present study, the feeding activity of
the epigeic earthworm, E. eugeniae was tested in
high-substrate column vermireactors by varying the
parameters such as: (i) substrate height, (ii) degree
of decomposition of the substrate, and (iii) mode of
substrate input. Neem leaves, which are locally
66
available in the experimental area, were used as
feeding substrate for the earthworms.
2. Materials and methods
Neem leaves collected from the Pondicherry
University campus and its vicinity were washed
with water to remove adhering particles. The
vermireactors of 35 cm width, 39 cm length and
100 cm height were prepared with basal layer of
coarse sand to a thickness of 3 cm followed by 5
cm thick layer of soil. Since the substrate height
was up to 60 cm in the experimental reactors, all
the four sides of the reactor were fabricated with
nylon mesh of pore size of 0.3 mm supported by a
wooden frame to avoid anaerobic condition set in.
Two sets of duplicate reactors were prepared in this
manner. In the first set, the reactors were filled with
neem leaves up to 45 and 60 cm height over sand
and soil bedding respectively (T1 and T2
treatments). In each reactor, 10 numbers of adult
worms of similar size were introduced after
determination of their biomass. The duration of
each run was 15 days. At the end of each run,
vermicast generated in the reactors were harvested
and the reactors were restarted with the leftover
substrate with adult worms, with which the reactors
were started. To maintain the initial substrate
height, fresh leaves were added to the left over
substrate whenever the height reduced. To maintain
the initial number of worms, hatchlings produced, if
any, were removed at the end of each run.
In order to study the influence of degree of
decomposition of substrate on epigeic preference
towards sand and soil, the second set of reactors
were prepared with 45 and 60 cm of substrate
height respectively (T3 and T4 treatments). At the
end of each run, the vermicast was harvested and
restarted with left over substrate from the previous
run. In these reactors, to study the influence of
increase in worm density on epigeic feeding
activity, juveniles and cocoons generated in the
previous run were reintroduced along with adult
worms after assessment. One set of reactors was
maintained as control, which was prepared with 45
cm height substrate and without soil and sand
bedding. The control reactor was also operated
similar to the second experimental set.
All the reactors were operated at identical
ambient conditions with temperature of 29 ± 4°C.
Moisture content of substrate was maintained about
60% throughout the study period by periodic
sprinkling of adequate quantity of water. At the end
of each run, the castings were carefully separated
from other particles by soft painting brush and
quantified. While disbanding the reactors, the adult
earthworms with which the experiment was started,
were washed, blotted dry and weighed without
emptying their gut to record their zoomass and
introduced back into the respective reactors. The
dead earthworms, if present, were replaced, while
starting the next run.
The presence of sand, soil and organic matter
in the vermicast was quantified by gravimetric
method. Samples oven dried at 105°C to a constant
weight were put in distilled water, and crushed to
release the soil/sand. The content was filtered
through a Whatman No. 42 filter paper. The residue
over the filter paper was kept in a muffle furnace at
550°C for 4 hours to remove organic matter present
in the castings (John, 2004). Petrographic thin
sections were made of air-dried vermicast using
Bueller PetroThin® thin sectioning system
(Buehler, Lake Bluff, IL, USA) after impregnating
them in araldite and xylene (solvent) mixture. The
resulting specimens were observed under polarizing
microscope to confirm the sand assimilation in the
vermicast (FitzPatrick, 1993).
The data was statistically analyzed by two-
way ANOVA to assess the impact of
presence/absence of sand and soil vermibed on
vermicast production, earthworm mortality,
zoomass gain and assimilation of sand and soil
content in the vermicast. The post hoc test LSD has
been done to find the significant difference between
each subject. All statistical analyses were
67
performed using the statistical software SPSS 16
package.
3. Results and discussion
The reactors T1, T2 and control were
operated for 12 runs by continuous feeding of
substrate at each run. In the T3 and T4 treatments,
substrate height of the reactors reduced to 5 cm
during the ninth run, hence these reactors were not
operated further. There was a significant difference
in the amount of sand and soil assimilated in the
vermicast with different treatments (p<0.001). The
soil assimilation rate was high in the initial runs
with all the treatments (Figure 1). In the first three
runs, the percentage of sand and soil content in the
reactors of T1 and T2 was 36.5 and 29.7% on
average, and those of T3 and T4 treatments were
28.4 and 24.0%, respectively. The high soil
assimilation rate in the initial runs was probably
due to presence of polyphenolic compounds in the
fresh neem leaves. A negative correlation between
the palatability of leaf litter and its total polyphenol
content of substrate demonstrated in previous
studies (Satchell and Low, 1967; Hendrickson,
1990; Tian et al., 2000), may be the reason for
earthworm preference towards sand and soil during
the initial runs. From the fourth run there was a
steady decline in assimilated sand and soil content
in the castings of both T1 and T2 treatments, which
might have attributed to the degradation of phenolic
components in the substrate. The previous study by
Gajalakshmi and Abbasi (2004) also support this
assumption, in which earthworms voraciously fed
the neem leaves after pre-composting.
In the T1 and T2 treatments, the sand and soil
assimilation rate was stabilized from the ninth run,
in which less than 5% of sand and soil content was
recorded with most of the cases. In the case of T3
and T4 treatments, the sand and soil assimilation
showed a declining trend up to fifth run and
subsequent runs showed increase in soil and sand
assimilation in the vermicast. Due to continuous
harvesting of vermicast at the end of each run, the
substrate height of these reactors steadily declined
throughout the experimental period, which might
have attributed to the high sand and soil
assimilation after fifth run. At ninth run, 33.3 and
17.4% of soil and sand assimilation was recorded in
the T3 and T4 treatments, and the substrate height
of these reactors were 6 and 9 cm, respectively at
the start of the ninth run. However, during the first
five runs, the sand and soil assimilation rate in the
Figure 1. Percentage of sand and soil particles entrapped in the castings of different treatments
0
10
20
30
40
3 4 5 6 7 8 9 10 11 12
Soil
and
san
d e
ntr
app
ed in
cas
tin
gs
(%)
Runs
T1
T2
T3
T4
68
Figure 2. Vermicast output, grams worm -1
day-1
recorded in reactors with different treatment.
T3 and T4 treatments were much lower than the T1
and T2 treatments. This indicates that the
earthworms did not prefer sand and soil, when the
degraded substrate is available to them. The initial
height of the substrate in the reactors did not show
any noticeable difference in sand and soil
assimilation in vermicast. The assimilation of sand
and soil in the castings was confirmed with
petrographic thin sections.
The different heights of the substrate did
not have any significant impact on vermicast
output, but the mode of substrate input (either one
time or continuous supply of substrate) has changed
the vermicast output considerably (Figure 2). But
the statistical analysis did not show any significant
difference within different treatments due to much
variation in data. During the first three runs, very
small quantity of vermicast was produced in all the
reactors. At the end of third week, the vermicast
output was 0.525 and 0.280 g worm-1
d-1
with T1
and T2 treatments, and those of T3 and T4 showed
0.805 and 0.215 g worm-1
d-1
, respectively. In
subsequent runs, there was increase in vermicast
output with all the treatments. The increase in
vermicast output was much higher in the T3 and T4
treatments during the first six runs; in some cases it
was about twofold higher than the T1 and T2
treatments. The reason for increase in vermicast
output in T3 and T4 treatments may be probably
due to increase in the degree of decomposition of
the substrate they contained. But, in the T1 and T2
treatments, the addition of fresh substrate at each
run to maintain the substrate height might have
increased the polyphenolic content of the whole
substrate which in turn reduced the vermicast
output. Studies on acacia (Acacia auriculiformis)
(Ganesh et al., 2009), oak, beech, larch, spruce
(Satchell and Low, 1967) and common St. John's
wort (Hypericum perforatum) (Schonholzer et al.,
1998) also shows that the freshly fallen leaves of
these plants are unacceptable by the worms due to
their phenolic and other toxic compounds. Further
runs, did not show any difference in vermicast
output with these treatments probably due to
acclimatization of the earthworms to this substrate.
Throughout the study, the vermicast output
from the control treatment was very close to the T1
treatment which comprised similar substrate height.
The changes in earthworm zoomass and mortality
also did not show any significant difference
between different treatments. During the first three
runs, there was loss in earthworm biomass and high
mortality was observed with all the treatments;
thereafter there was no significant difference in any
0
0.5
1
1.5
2
2.5
3 4 5 6 7 8 9 10 11 12
Ver
mic
ast
ou
tpu
t, g
wo
rm -1
d-1
Runs
T1
T2
T3
T4
Control
69
of these parameters. The overall results show that
existence of sand and soil bedding neither
influenced the vermicast output nor their growth or
mortality with neem as a feed.
4. Conclusions
The findings of the present study clearly
shows that the diet of the epigeic worm E. eugeniae
consist of considerable portion of soil and sand in
the initial period. However, the amount of soil and
sand ingestion rate drastically reduced as the days
progressed. The earthworm adaptability to neem
leaves might have reduced the sand and soil
ingestion rate. Varying substrate height of the
reactors did not show any influence on mineral soil
ingestion, provided the height was not less than 5
cm. However, ingestion of soil and sand particles
neither enhanced the E. eugeniae’s digestion
process nor feeding rate or its growth as reported
with anecic and endogeic earthworms.
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70
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EFFECT OF SAND AND S
PHYTOPHAGOUS EARTHWO
EUGENIAE ON THE PHYSICAL AND
PROPERTIES OF VERMIC
EFFECT OF SAND AND SOIL INGESTION BY
PHYTOPHAGOUS EARTHWORM EUDRILUS
ON THE PHYSICAL AND CHEMICAL
PROPERTIES OF VERMICAST
OIL INGESTION BY
EUDRILUS
CHEMICAL
Chapter
5
71
A paper based on this chapter has been
submitted for publication.
CChhaapptteerr 55
Effect of sand and soil ingestion by phytophagous earthworm
Eudrilus eugeniae on the physical and chemical properties of
vermicast
Abstract
The studies reported in chapters 3 and 4 have revealed that even if sand/soil are not required for the epigeics
growth and survival, the earthworms prefer sand/soil with fresh phytomass. This tendency subsided once the
earthworms got acclimatized to the feed. However, the effect of sand and soil ingestion on the properties of
vermicast is not yet explored. Hence, a controlled experiment was carried out with Eudrilus eugeniae in
vermireactors with different masses of substrate, to study the feeding behavior of epigeics and its impact on
the properties of vermicast they generate. The results reinforce our earlier finding that the epigeics ingest soil
and sand inspite of the luxury availability of phytomass, but the consumption of sand and soil neither
facilitate their survival, growth and fecundity of the epigeics. Ingestion of sand and soil had increased the
bulk density and particle density of the vermicast in addition to reduction of pore space, water retention
capacity, and nutrient content.
1. Introduction
Of all the larger inhabitants of the soil,
probably none is more important than the
earthworm in terms of soil formation and
maintenance (Carson, 1962). The earthworms
significantly affect the soil structural characteristics
and distribution of resources to many soil animals
and plants mainly through the castings they
produce. The castings exists in various forms and
features which is determined by earthworm species,
their habitat and feeding behavior. Generally the
castings are enriched in organic matter, nutrients
and foster high levels of microbial activity due to
the selective foraging of organic particles (Fonte et
al., 2007). In addition, anecic and endogeic
earthworm groups defined by Bouche (1977),
ingest considerable portion of soil/sand along with
organic matter and produce complex organo-
mineral structures as castings. Although, ingestion
of soil/sand particulate by earthworms facilitate
mechanical fragmentation of organic matter during
gut transition (Schulmann and Tiunov, 1999),
assimilation of soil particles in castings alter
surface soil texture and other physical properties
(Lavelle and Spain, 2001; Blanchart et al., 2004;
Jouquet et al., 2008a,b), microbial activity and
diversity (Marhan, 2004), and therefore the
mineralization or sequestration of soil organic
matter and the retention or the leaching of mineral
nutrients (Brady and Weil, 1999; Lal, 2004).
Although a number of studies have discussed
these aspects, the information are limited to anecic
and endogeic groups of earthworms. Even though,
the epigeics are reported to ingest soil and sand in
few studies (Martin, 1982; Bostrom, 1988;
Karthikeyan et al., 2014), less is known about the
impact of soil ingestion on their digestion, nutrient
uptake and properties of vermicast. Studying this
aspect is particularly important to understand the
role of epigeics in soil processes as well as to
enhance the nutrient cycle of organic matter in both
agricultural and waste management systems.
72
Therefore, in the present study, an attempt has been
taken to understand the impact of feeding activity
of the epigeic earthworm, Eudrilus eugeniae on
their growth and properties of vermicast produced
by them. The earthworms were grown with neem
leaves as feed, in the presence/absence of soil and
sand with different quantities of the substrate. The
presence of sand and soil in the vermicast was
confirmed by petrographic thin section method. The
amount of soil and sand assimilated in the castings
was quantified by both gravimetric and wavelength
dispersive X-ray fluorescence spectrometry
methods. The effect of soil/sand ingestion on the
growth of earthworm and vermicast production had
been monitored in the neem fed vermireactors for
240 days. The difference in physical and chemical
properties of vermicast generated from reactors
supplied with or without soil/sand was also studied
and the results are discussed in brief.
2. Materials and methods
2.1. Experimental design
Neem leaves were collected from the
Pondicherry University campus and its vicinity.
The leaves were washed with water to remove
adhering particles. Rectangular, 41 liter wooden
boxes (30 cm high with surface area of 35 x 39 cm)
were used as vermireactors. They were lined up
with thick transparent plastic sheets to make
reactors impermeable, prevent earthworms escaping
from the reactors and also to protect them from
predators. Four sets of duplicate vermireactors were
employed. First and second set of duplicate reactors
were filled with 250 and 500 g dry weight
equivalent of neem leaves (T1 and T2 treatments).
The third and fourth sets of reactors were prepared
with basal layer of coarse sand to a thickness of 3
cm followed by a 5 cm thick layer of soil. Over it
250 and 500 g dry weight equivalent of neem
leaves was placed (T3 and T4 treatments). Ten
adult earthworms having approximately equal size,
randomly picked from their cow dung-based
culture, were introduced in each of the reactors.
2.2. Vermireactors operation
All the reactors were operated at identical
ambient conditions with a temperature range of
29±4°C. Moisture content of substrate was
maintained about 60% throughout the study period
by periodic sprinkling of adequate quantity of
water. The duration of each run was 15 days. At the
end of each run, vermicast mass, mortality and
changes in the earthworm zoomass were
determined, and then the reactors were restarted
with the leftover substrate with adult worms, with
which reactors were started. To maintain the initial
number of worms, hatchlings produced, if any,
were removed and dead worms were replaced,
while starting the next run. During the first trial, the
reactors were operated for 135 days. The second
trial comprised of 105 days, which had been
operated with newly assembled reactor contents
and earthworms from the previous trial.
2.3. Analytical methods
Moisture content of castings was determined
by weight loss at 105°C. The bulk density and
particle density of samples was determined
according to Bashour and Sayegh (2007). To
measure the water holding capacity, the samples
were filled in cylinders with a perforated base and
immersed in water and drained. The quantity of
water taken up by samples is determined by drying
to constant mass at 105°C (Margesin and Schinner,
2005). The total porosity and water filled pore
space (WFPS) were calculated from the particle and
bulk density values of the respective samples
(Carter and Gregorich, 2008). The electrical
conductivity (EC) was measured with sample
suspension of 1:2 (w/v) by using EI™ 611E EC
meter (Bashour and Sayegh, 2007).
Total organic (Corg) content in the castings
was measured by modified dichromate redox
method (Heanes, 1984). In this method, external
heating was applied during the oxidation process in
order to quicken and complete oxidation of Corg in
73
the sample. Total nitrogen (Ntot) content was
determined by modified Kjeldahl method
(Kandeler, 1993) using Kel Plus™ semi-automated
digester and distillation units (Elico Ltd,
Hyderabad, India). In order to include nitrate,
nitrite, nitro and nitroso groups, a mixture of
salicylic acid and sulfuric acid was used for
digestion. The SiO2, potassium and phosphorus
content in the vermicast were determined by
Bruker™ S4-Pioneer model wavelength dispersive
X-ray fluorescence spectrophotometer (WD-XRF,
Bruker, Billerica, MA, USA). The samples were
ground to particle size well below 100 µm using
ball mill in order to minimize the grain size
interference on XRF-measurement.
2.4. Assessment of soil/sand content in the
vermicast
The presence of sand, soil and organic matter
in the vermicast was quantified by gravimetric
method. Samples oven dried at 105 °C to a constant
weight were put in distilled water, and crushed to
release the soil/sand. The content was filtered
through a Whatman No. 42 filter paper. The residue
over the filter paper was kept in a muffle furnace at
550°C for 4 hours to remove organic matter present
in the castings (John, 2004). Petrographic thin
sections were made of air-dried vermicast using
Bueller PetroThin® thin sectioning system
(Buehler, Lake Bluff, IL, USA) after impregnating
them in araldite and xylene mixture. The resulting
specimens were observed under a polarizing
microscope to confirm the sand assimilation in the
vermicast (FitzPatrick, 1993).
2.5. Data analysis
The influence of sand and soil ingestion on
the growth and survival of earthworm, vermicast
mass and its physico-chemical properties were
assessed by independent sample t-test. The
statistical calculation was carried out using SPSS
16 software.
3. Results and discussion
3.1. Assimilation of sand/soil in the vermicast
The percentage of sand and soil entrapped in
the earthworm castings in each run of 15 days
duration with different treatments is shown in
Figures 1 and 2. There was a significant difference
in the amount of sand/soil assimilation in the
vermicast from the reactors with different amount
of substrate and duration of reactor operation (p
<0.001).
Figure 1. Percentage of sand and soil entrapped in the vermicast from reactors consisting sand/soil with 250
g (T3a and T3b, duplicates) and 500 g substrate (T4a and T4b, duplicates) in the first trial.
0
15
30
45
60
75
3 4 5 6 7 8 9
Soil
and
san
d e
ntr
app
ed in
VC
(%
)
No. of runs
T3 a T3 b T4 a T4 b Linear (T3 b)
74
Figure 2. Percentage of sand and soil entrapped in the vermicast from reactors consisting sand/soil with 250
g (T3a and T3b, duplicates) and 500 g substrate (T4a and T4b, duplicates) in the second trial.
In both the first and second trials, high sand/soil
assimilation was recorded during the initial run
with all the treatments. During the first three runs,
the percentage of sand and soil content in the
reactors of T1 was 60.7% on average, and those of
T2 treatments were 47.0%, respectively. There was
a steady decline in sand/soil assimilation with all
the treatments up to 8th and 6
th run of first and
second trials, respectively. The high sand/soil
assimilation in the initial run was probably due to
the presence of some of the unaccepted substance
in fresh neem leaves like phenolic and other toxic
compounds. Previous reports on leaves of many
tree species, includes oak, beech, larch, and spruce,
which are reported to contain phenolic compounds,
were also shown unpalatable by earthworms
(Satchell, 1967). Similarly, studies reported by
Hendriksen (1990) and Schonholzer et al. (1998)
revealed that the palatability of different kinds of
leaf litter by Lumbricus spp. and Aporrectodea spp.
is largely determined by the C:N ratio, lignin and
the polyphenol concentration of leaf litter.
Reduction in the assimilation of sand/soil in the
vermicast in further runs indicates that degradation
of the unacceptable substance in fresh leaves has
been initiated. The percentage of sand/soil
assimilation was reduced to about 20% in the 8th
and 6th run with first and second trials, respectively.
The percentage of SiO2 in the vermicast
significant by varied with different reactors
consisting different amounts of substrate (Table 1).
The percentage of SiO2 was 40.4% in the castings
from T1 treatment, and those of T2 were 24.4%,
respectively. In the first trial, the gravimetrically
determined sand/soil content in the castings was
higher in reactors of T1 treatment than those of T2
treatment during the first eight runs. From ninth run
onwards there significant difference in amount of
sand/soil assimilated in the castings from different
treatments. During the second trial, the assimilation
of sand/soil in the castings had stabilized on the
sixth run probably due to acclimatization of the
earthworms to this substrate. However, all the
reactors were showing slight increase in soil/sand
assimilation in last few runs. Due to continuous
harvesting of vermicast at the end of each run, the
substrate height of these reactors steadily declined
drastically, which might have attributed to the high
sand/soil assimilation in last few runs.
0
15
30
45
60
75
2 3 4 5 6 7
Soil
and
san
d e
ntr
app
ed in
VC
(%
)
No. of Runs
T3 a T3 b T4 a T4 b Linear (T3 b)
75
3.2. Vermicast output
Presence of sand/soil bedding showed
significant influence on vermicast production, but
the difference in the substrate quantities did not
show any noticeable changes in vermicast output
(Figure 3). During the initial runs, there was very
less quantity of vermicast production with all the
reactors. At the end of third week, the vermicast
output was 0.318±0.046 g worm-1
day-1
with
reactors without bedding, and that of reactors with
bedding showed 0.482±0.048 g worm-1
day-1
.
Fourth and fifth runs showed an increase in
vermicast output with all the treatments, and it was
about fivefold in some of the reactors. The drastic
increase in vermicast output may be due to increase
in the degree of decomposition of the substrate they
contained. As discussed in the previous section, the
degradation of polyphenolic and other toxic
compounds in the neem leaves would have
increased the vermicast output in all the treatments.
The vermicast output in subsequent runs, did
not show any significant difference while the height
of the substrate was not less than 1 cm. But in all
the runs, the production of vermicast was higher in
the reactors with bedding than without bedding
which indicates that the presence of bedding had
significant influence on vermicast output. However,
the vermicast from the reactors with bedding
consisted of a considerable portion of sand/soil.
There was no significant difference observed with
both trails, when the vermicast output of reactors
without bedding was compared with reactors
consisting bedding after subtracting the amount of
assimilated sand/soil with vermicast output. This
indicates that the ingested amount of substrate
alone by earthworms was not influenced by the
presence of bedding.
3.3. Growth and survival of earthworms
The presence of bedding and amount of
substrate input in vermireactors did not show any
significant influence on the mortality and changes
in earthworm zoomass. The first two runs of first
trial showed reduction in zoomass in most of the
reactors; whereas in the second trial, the zoomass
reduction was observed only in the first run. In the
first trial, the reduction in zoomass was
0.036±0.010 and 0.026±0.012 g worm-1
, and in the
Figure 3. Vermicast output, grams worm -1
day-1
(mean ± SD) recorded in reactors without sand/soil + 250
and 500 g substrate (T1 and T2 treatments) and reactors consisting sand/soil + 250 and 500 g substrate (T3
and T4 treatments).
0
1
2
3
a b a b a b a b
T1 T2 T3 T4
Ver
mic
ast
ou
tpu
r g
wo
rm-1
d-1
Treatments
1st Trial
2nd Trial
Linear (2nd Trial)
76
second trial, 0.039±0.022 and 0.050 ±0.017 g
worm-1
at the end of the first run in the reactors with
the presence and absence of bedding, respectively.
The subsequent runs of both trials showed increase
in zoomass in most of the cases. Likewise, high
earthworm mortality was also recorded in the initial
runs with both the trials. In the first trial, the
reactors without bedding showed a maximum of
50% mortality with T3 treatment. The reactors with
bedding showed a maximum mortality of 40% in
the T2 treatment. Similarly, in the second trial, a
maximum of 30% mortality was observed in one of
the reactors without bedding, and that of a reactor
with bedding showed 20% mortality, respectively.
Except the last run of both the trials, there was no
significant mortality. The last run of both trials
showed reduction in zoomass and mortality in some
of the reactors, probably due to decline in substrate
availability in these reactors. Similarly, increase in
the mortality of earthworms with few or no organic
residues added soil has been already reported in
both pot and field trials (Edwards, 2004).
3.4. Physical and chemical properties of
vermicast
The physical properties of vermicast significantly
varied (p <0.001) with presence or absence of
bedding. In the reactors without bedding, the
amount of substrate had no impact on the properties
of vermicast, but it exhibited significant influence
when the reactors consisted bedding. The presence
of bedding increased the bulk and particle density,
and reduced the total porosity, WFPS and WHC of
the vermicast (Table 1). The average bulk density
of castings from the reactors of both T3 and T4
treatments was 0.303 g cm-3
, whereas it was 0.672
and 0.467 g cm-3
in the T1 and T2 treatments,
respectively. In the case of particle density, the T3
and T4 treatments showed 1.145 g cm-3
, and those
that of T1 and T2 was 1.706 and 1.441 g cm-3
,
respectively. Assimilation of high density soil
particle in the vermicast might be the reason for
higher bulk and particle density of castings from
reactors comprised of bedding. Studies on anecic
earthworms, Martiodrilus carimaguensis and
Lumbricus terrestris with different soil types have
also shown increase in density of castings when the
mineral soil consumption was high (Decaëns, 2000;
Josehko et al., 1989). Fluctuation in the bulk and
particle density of castings from the reactors
consisting different quantities of substrate were
probably due to the variation in sand/soil
assimilated by the earthworms in these reactors.
The total porosity and WFPS of castings were also
Table 1. The physical properties of vermicast generated from the reactors without sand/soil + 250 and 500 g
substrate (T1 and T2 treatments) and reactors consisting sand/soil + 250 and 500 g substrate (T3 and T4
treatments). The alphabets ‘a’ and ‘b’ represent duplicate reactors.
Treatments Water Content
%
Particle Density
g cm-3
Bulk Density
g cm-3
Pore Space
% WFPS % WHC %
EC
mmhos cm-1
T1-a 72.9±1.8 1.139±0.004 0.288±0.006 74.7±0.5 24.7±1.2 525.8±10.9 2.20±0.05
T1-b 75.4±1.6 1.192±0.007 0.309±0.006 74.1±0.4 26.4±0.3 590.5±12.9 2.23±0.04
T2-a 68.0±2.5 1.127±0.008 0.303±0.006 73.2±0.6 25.0±1.2 614.4±11.4 2.16±0.03
T2-b 73.2±1.5 1.122±0.007 0.312±0.009 72.2±0.7 28.2±1.5 531.8±21.5 2.32±0.10
T3-a 33.2±0.4 1.709±0.018 0.709±0.013 58.5±0.3 40.1±1.2 204.3±27.3 1.66±0.16
T3-b 35.4±0.5 1.703±0.017 0.634±0.008 62.8±0.4 35.6±0.8 232.5±7.3 1.76±0.03
T4-a 42.0±0.9 1.511±0.013 0.496±0.014 67.2±1.0 30.8±1.2 350.7±6.7 1.69±0.03
T4-b 47.3±0.3 1.371±0.021 0.438±0.017 68.0±0.7 30.3±1.7 389.6±8.0 1.82±0.04
77
Table 2. The chemical properties of vermicast generated from the reactors without sand/soil + 250 and 500 g
substrate (T1 and T2 treatments) and reactors consisting sand/soil + 250 and 500 g substrate (T3 and T4
treatments). The alphabets ‘a’ and ‘b’ represent duplicate reactors.
Treatments TC mg g-1
TN % TK % TP % SiO3 %
T1-a 390.9±9.3 31.87±0.09 1.185±0.044 0.527±0.008 0.114±0.012
T1-b 388.0±4.3 34.06±0.11 1.180±0.046 0.473±0.011 0.092±0.005
T2-a 396.8±5.1 33.78±0.12 0.987±0.067 0.479±0.004 0.119±0.007
T2-b 389.9±3.5 33.32±0.04 1.032±0.046 0.521±0.005 0.099±0.008
T3-a 150.5±3.8 12.15±0.09 0.781±0.025 0.349±0.041 39.26±5.59
T3-b 139.8±7.7 15.33±0.20 0.705±0.017 0.428±0.026 41.47±6.66
T4-a 187.4±9.3 15.31±0.17 0.761±0.024 0.453±0.033 26.33±4.96
T4-b 186.8±7.5 14.87±0.15 0.842±0.017 0.365±0.029 22.54±2.70
significantly lower in the reactors with bedding
than the reactors devoid of bedding. The total
porosity of castings generated from the T1 and T2
treatments was 18.5 and 7.0% lower than the T3
and T4 treatments, respectively. Similarly castings
generated from the T1 and T2 treatments showed
about 60 and 34%, respectively lower WHC than
the castings from corresponding reactors without
soil/sand bedding. The lower WHC of the castings
in reactors comprised of bedding may be attributed
by the lower pore space of the castings (Chaudhuri
et al., 2009). In these treatments, the existence of
soil/sand particle in the castings reduced the
porosity and WHC of vermicast.
There was significant influence (p < 0.001)
on the chemical properties of vermicast with
reactors consisting of bedding. But the amount of
substrate had no impact on the properties of the
vermicast. The castings from the reactors without
bedding had maximum total C, N, P and K content
than the castings from reactors comprising of
bedding (Table 2). The castings from T3 and T4
reactors showed about 63 and 52% higher organic
carbon than the T1 and T2 reactors, respectively.
Similarly the nitrogen content was about 58 and
54% lower in the T1 and T2 reactors compared to
T3 and T4 reactors. The total potassium content of
castings was approximately 37% lower in T1
reactors than T3 reactors, and those of T2 reactors
showed 20% lower than T4 treatment, respectively.
A significant amount of sand and soil entrapped in
the castings may be the reason for the lower content
of C, N and K in castings from reactors with
sand/soil bedding compared to the ones without
bedding. Phosphorus content of castings has not
showed any significant difference between different
treatments. The phosphorus present in the soil/sand
entrapped in the castings might have attributed this
insignificant difference between these treatments.
4. Conclusions
The study gives inference that epigeic
earthworm – E.eugeniae also ingest sand and soil
like anecics and endogeics species. But once the
earthworms got acclimatized to the phytomass as
feed, the amount of soil and sand ingestion was
insignificant. Moreover, the ingestion of sand and
soil by epigeics, neither facilitated their digestion
nor its survival as there is no significant difference
in zoomass gain and earthworm mortality in
reactors with and without sand/soil bedding. There
was increase in the bulk density and particle
density, reduction in the pore space, water holding
capacity and nutrient content of vermicast in
reactors with soil/sand bedding. The overall finding
shows that the epigeic earthworm, E.eugeniae
78
really does not require sand/soil in their diet unlike
anecic and endogeic species. Moreover, the
physico-chemical characterization of vermicast
reveals that the assimilation of sand/soil in the
vermicast may reduce their beneficial impact on
plant growth and soil.
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80
EFFECT OF VERMICAST GENERATED FROM AN
ALLELOPATHIC WEED LANTANA (LANTANA CAMARA)
ON SEED GERMINATION, PLANT GROWTH, AND
YIELD OF CLUSTER BEAN (CYAMOPSIS
TETRAGONOLOBA)
Chapter
6
81
A paper based on this chapter has been published in
Environmental Science and Pollution Research DOI: 10.1007/s11356-014-3103-5
CChhaapptteerr 66
Effect of vermicast generated from an allelopathic weed
lantana (Lantana camara) on seed germination, plant growth,
and yield of cluster bean (Cyamopsis tetragonoloba)
Abstract
In perhaps the first-ever study of its kind the effect of vermicompost, derived solely from an allelopathic
weed, on the germination, growth, and yield of a botanical species has been carried out. In test plots, the soil
was treated with the vermicompost of lantana (Lantana camara) at the rates of 5, 7.5 and 10 tons per hectare
(t ha-1
) and cluster bean (Cyamopsis tetragonoloba) was grown on it. The performance of these systems was
compared with the systems in which the soil was fortified with inorganic fertilizers (IF) in concentrations
equivalent to those present in the respective vermicompost (VC) treatments. Additionally a set of control was
studied in which the soil was used without fortification by either VC or IF. It was seen that up to 51.5%
greater germination success occurred in the VC treatments compared to controls. VC also supported better
plant growth in terms of stem diameter, shoot length, shoot mass, number of leaves, and leaf pigments. The
positive impact extended up to fruit yield. In addition, vermicast application enhanced root nodule formation,
reduced disease incidence, and allowed for a smaller number of stunted plants. The results indicate that
allelopathic ingredients of lantana seem to have been totally eliminated during the course of its
vermicomposting, and that lantana vermicompost has the potential to support germination, growth and fruit
yield better than equivalent quantities of inorganic fertilizers.
1. Introduction
Lantana (Lantana camara) is among the
world’s most noxious weeds (GISD, 2014). It is a
shrub, native to South America (Ghisalberti, 2000;
Henderson, 2001), and belongs to the family of
Verbenaceae which comprises of about 650 species
occurring in over 60 countries including India
(Sharma et al., 2005; Kohli et al., 2009; Hiremath
and Sundaram, 2013). Lantana grows in a wide
range of environmental conditions, infesting
millions of hectares of natural ecosystems and
cultivated lands, causing great damage to
biodiversity (Gentle and Duggin, 1997; Batianoff
and Franks, 1998; Day et al., 2003; Vardien et al.,
2012). It possesses mammalian toxicity and is
known to induce photodermatitis, jaundice, liver
damage and death in animals when they graze on its
leaves (Sharma et al., 1988, Pass, 1991; Bevilacqua
et al., 2011). It is also strongly allelopathic, having
several compounds which repel or toxify other
vegetation, thereby preventing their growth (Holm
et al., 1991; Gentle and Duggin, 1997; Day et al.,
2003; Ahmed et al., 2007).
Substantial efforts have been made, and are
continuing across the world, with billions of dollars
having been invested, to control invasives like
lantana by physical, chemical or biological means
(Erasmus and Clayton, 1992; McFadyen, 1998;
Day et al., 2003; Zalucki et al., 2007). However,
these attempts have not succeeded in even
82
controlling, let alone eradicating, any of the major
weeds. It appears that if processes can be developed
for gainfully utilizing the invasives, it may not only
offset the costs of mechanically removing them but
also exercise some control over their spread. One of
the possible options is conversion of weeds into
vermicompost and utilizing the latter as a soil
fertilizer.
Vermicomposting of a substrate is believed to
convert some of its nutrients into more bioavailable
forms and bestow upon the substrate microflora that
is beneficial for soil health (Gajalakshmi and
Abbasi, 2008; Edward et al., 2011). Additionally
hormones, enzymes and pest–repellants are
believed to be added to the substrate as it passes
through the earthworm gut and gets converted to
vermicast. Among these and several other attractive
features of the vermicomposting option is its ability
to sequester nearly all the carbon that is contained
in the substrate-to-be-vermicomposted (Abbasi et
al., 2011; Banupriya et al., 2014). But, despite the
great potential of vermicomposting as a phytowaste
utilization option, its use in handling phytomass in
general and weeds in particular has yet to begin
(Nayeem-Shah, 2014).
Recent reviews (Nayeem-Shah, 2014;
Nayeem- Shah et al., 2014) reveal that the main
constraint which has prevented the use of
vermicomposting in processing large quantities of
phytomass has been the incapability of the
conventional vermicomposting technology, which
is primarily geared to the utilization of animal
manure, in handling phytomass. To circumvent this
hurdle various authors have been, as reviewed by
Nayeem-Shah (2014), pre-composting the weeds
for 3 weeks or more, then subjecting the pre-
compost to vermicomposting after blending the
substrate with animal manure. Another 2-3 months
or more pass before the vermicomposting is
deemed to have completed. But the reliance on pre-
composting, animal manure, and the very long
overall processing time make the system
thoroughly unwieldly and uneconomical. The result
is that despite a spurt in laboratory- scale studies on
the vermicomposting of phytomass in recent years
(Nayeem-Shah, 2014), there is no attempt, nor it
appears possible, to utilize conventional
vermicomposting technology in processing
phytomass.
It has also been a matter of concern whether
phytomass-based vermicompost will be as
beneficial to soil and plants as manure-based
vermicompost is. This concern is particularly
relevant for plants like lantana which are known to
possess constituents that are toxic to animals and
other species of plants.
A solution to the first of the problems has
now emerged as a result of the efforts of S.A.
Abbasi and coworkers who have developed the
concept of high-rate vermicomposting and
associated technology (Gajalakshmi et al., 2002,
2005; Abbasi et al., 2009, 2011; Tauseef et al.,
2013). As detailed elsewhere (Nayeem-Shah et al.,
2013, 2014) the high-rate vermicomposting
technology makes it possible to directly
vermicompost phytomass; that, too, at a rapid rate.
Using this technology, it has also been possible to
directly vermicompost lantana and obtain its
vermicast in an efficient and convenient manner
(Kumar et al., 2012). With the prospect of
inexpensively and efficiently vermicomposting
lantana now assured, it has become imperative to
address the other major concern-the fertilizer value
of lantana vermicompost. The present paper is
devoted to this concern.
2. Materials and methods
The studies were conducted at Pondicherry
University, Puducherry, India, located on the east
coast of the Indian peninsula (11°56’N, 79°53’E).
This region experiences hot summers during March
– July (maximum day temperature 35-38°C), and
mild winters during December - February
(maximum day temperature 29-32°C). The average
annual rainfall is about 1300 mm, concentrated
83
mainly during October – December but with a few
rainy days occurring in July–August and January as
well. For vermicomposting, reactors fabricated with
aluminum sheet of 140 liter volume were employed
for direct vermicomposting of lantana (Kumar et
al., 2012). The periodically harvested vermicast
was stored in sealed plastic containers. The study
on germination and growth was conducted outdoors
in 49 liter volume containers (40 cm height with
surface area 35 x 35 cm), lined with high-density
polyethylene (HDPE) sheets. The soil used in the
experiments was collected from a previously
uncultivated piece of land so that the results are not
influenced by any earlier fertilizer application. The
physico-chemical properties of the vermicast and of
the soil with which it was used are given in Table 1.
The experiments were carried out during July –
October which is ideal for growing cluster beans in
the study area (ICAR, 2011).
For studying the impact of vermicast on
cluster bean, three sets of experiments were
conducted. Based on the nitrogen, phosphorus, and
potassium (NPK) concentrations seen in the
vermicompost (Table 1), and the levels of NPK
needed in the soil for cluster bean (ICAR, 2011), it
was calculated that the cluster bean would require
7.5 t ha-1
of lantana vermicompost (VC) for its
optimum growth. In the first set, one batch of
containers was supplied with 7.5 t ha-1
of VC while
two other batches were given 5 and 10 t ha-1
of VC.
These doses were calculated on the basis of surface
area of the containers, and were 123, 92, and 61 g
for 10, 7.5 and 5 t ha-1
treatments, respectively.
Second set comprised of chemical fertilizers (IF)
which were treated with nutrients N, P, K, Ca, Mg,
S, Fe, Mn, Cu, Zn, B, Mo and Cl in concentrations
equivalent to those present in the respective
vermicompost treatments (Table 2). The nutrients
were supplied in two installments: half at the time
of sowing and the rest at the onset of flowering.
The last set had no supplementation of nutrients
and served as control. In this manner 3 sets of
experiments, encompassing seven treatments were
carried out. As each treatment had 36 containers,
each with one plant, 252 containers were utilized
for the main experiment. Another 150 containers,
30 of control and 20 each of the doses of VC and IF
treatments that were employed, were maintained as
spares. These were used to replace the containers of
which plants were sacrificed for analysis, or in
which the plants happened to have died. The
instances of plant death were very few, less than
3%. The Pusanavabahar variety cluster bean, which
is locally available, was used. Following the
assessment of germination over eight days, growth
and yield were monitored for an additional three
months. Throughout the experiments, adequate
watering was performed to maintain 20-30% (v/w)
moisture. Deweeding was done periodically. Neem
extract was applied as a mild pesticide in a few
instances when pests were seen infesting the plants.
2.1. Germination, plant growth and yield
characteristics
Seed germination was assessed for 8 days
from the day of sowing and was quantified in terms
of germination value (GV) using the formula of
Djavanshir and Pourbeik (1976):
Where GV is germination value; DGS is daily
germination speed which is computed by dividing
cumulative germination percent by the number of
days elapsed since the beginning of the test; GP is
germination percentage at the end of the test, and N
is frequency or number of DGS that are calculated
during the test. The unit for GV is germinated
seeds/day. It is used as an index to statistically
assess the effects of different treatments.
The growth was assessed on the basis of stem
diameter, length of shoot and root, number of
leaves, number of nodules present in the root, the
nodules size and shoot/root biomass for 3 months.
Plants were harvested periodically, and at each
harvest, the fruits were counted and their fresh and
84
Table 1. Chemical and physical properties of vermicast
and soil used in the study
BDL – Below detection limit
dry weights were determined. To quantify biomass
in terms of dry weight, samples were oven-dried at
85°C to constant weight. Chlorophyll a and b, and
carotenoids were extracted by the method described
by Moran and Porath (1980). One gram of mature
leaf was ground and incubated with N,N-di-methyl
formamide (DMF) for 24 h at 4 °C in dark, shaking
every 6 hours (Moran and Porath, 1980). The
resultant supernatant was read at 470, 647 and 664
nm and the concentration of pigments were
determined as detailed by Wellburn (1994). The
yield was calculated in the form of harvest index
(HI) which is the ratio of weight of beans per plant
to the above-ground biomass. The yield as reflected
in pod size, pea size, number of peas per pod and
diseased pods, if any, were recorded at each harvest
with randomly selected 50 fruits.
During the course of the experiment, the
plants were infected with bacterial blight
(Xanthomonas campestris) and alternaria blight
(Alternaria spp). Infestation of white fly (Bemisia
tabaci) also occurred. The bacterial blight infection
was seen in the symptoms of black leaf spots,
necrotic lesions at leaf tips, black streaks on
petioles and stems, split stems, and defoliation
(Mihail and Alcorn, 1985; Ren, 2014). Plants were
considered infected by Alternaria spp. when there
were dark brown lesions on leaves with concentric
zonations demarcated with light brown lines. In
severe infections, several spots merged and gave
the leaves a blighted look, eventually causing
defoliation (Orellana and Simmons, 1966;
Yogendra et al., 1995). The number of plants that
died, either with or without any symptoms of
infection, and stunted plants were also recorded.
2.3. Analytical methods
Total organic carbon was measured by
modified dichromate redox method according to
Heanes (1984). Total nitrogen content was
determined by the modified Kjeldahl method
(Kandeler, 1993) using Kel Plus™ semi-automated
digester and distillation units. Inorganic N - NH4+
and NO3- were extracted in 2M KCl solution (1:10
weight: volume) and determined by modified
indophenol blue and Devarda’s alloy methods,
respectively (Jones, 2001; Bashour and Sayegh,
2007). Extractable potassium, calcium and sodium
were determined using a Flame photometer
(Elico™ CL378) after extraction with neutral 1N
ammonium acetate solution (Carter and Gregorich,
2008). Extractable magnesium, boron, copper, iron,
manganese, zinc, molybdenum were determined
using a Jobin Yvon – Ultima 2 model inductively
coupled plasma atomic emission spectroscopy (ICP
Variables Concentration
Vermicast Soil
Chemical properties
pH 6.47±0.01 6.30±0.10
Total organic carbon g kg-1
330.3±4.3 8.87±0.02
Total Nitrogen g kg-1
18.4±0.1 2.66±0.02
Plant available form of
Phosphorus mg kg -1
80.7±0.9 0.41±0.01
Potassium g kg-1
5.73±0.03 0.40±0.00
Sulphur mg 100g-1
1.34±0.01 0.54±0.01
Calcium g kg-1
5.49±0.02 8.27±0.01
Magnesium g kg-1
5.77±0.05 0.09±0.02
Boron mg kg-1
46.2±0.8 26.9±1.2
Copper mg kg-1
44.2±3.9 5.08±0.15
Iron mg kg-1
50.6±2.0 59.9±2.9
Manganese mg kg-1
227.2±8.4 45.1±2.2
Zinc mg kg-1
162.4±6.0 55.0±2.6
Molybdenum mg kg-1
BDL BDL
Physical properties
Dry weight % 43.1±0.3 94.7±0.1
Bulk density g cm-3
0.40±0.00 1.28±0.00
Particle density g cm-3
1.35±0.01 2.70±0.12
Water-holding capacity % 248.0±10.2 36.9±2.4
Electrical conductivity
mmhos cm-1
9.36±0.01 0.12±0.02
Total porosity % 70.6±0.2 53.1±2.0
Air filled porosity % 48.1±0.1 46.5±2.1
85
Table 2. Amount of inorganic fertilizer applied equivalent to vermicast treatment
Nutrients Form in which
applied
Mass %
of nutrient
in applied
compound
10t ha-1
7.5t ha-1
5t ha-1
Concentra
tion of
nutrient in
VC
(kg ha-1
)
Amount
of IF
applied
(kg ha-1
)
Concentration
of nutrient in
VC
(kg ha-1
)
Amount
of IF
applied
(kg ha-1
)
Concentration
of nutrient in
VC
(kg ha-1
)
Amount
of IF
applied
(kg ha-1
)
Nitrogen CH4N2O 46.65 66.26 142.1a 49.70 106.5
a 33.13 71.03
a
Phosphorus (NH4)2HPO4 23.45 34.79 148.3 26.09 111.3 17.40 74.17
Potassium KCl 52.44 24.82 47.32 18.61 35.492 12.41 23.66
Sulphur 0.058 Nilb 0.043 Nil
b 0.029 Nil
b
Calcium CaCO3 40.04 23.70 59.19 17.78 44.39 11.85 29.59
Magnesium MgO 60.30 24.91 41.31 18.68 30.98 12.45 20.65
Boron Na2B4O7.10H2O 11.34 0.199 1.756 0.149 1.317 0.100 0.878
Copper CuSO4.5H2O 25.45 0.191 0.749 0.143 0.562 0.095 0.375
Iron FeSO4.7H2O 20.09 0.218 1.086 0.164 0.815 0.109 0.543
Manganese MnSO4. H2O 32.50 0.980 3.015 0.735 2.262 0.490 1.508
Zinc ZnCl2 47.97 0.700 1.460 0.525 1.095 0.350 0.730 a The total amount of N applied in the form of urea and di-ammonium phosphate is equal to the concentration of N applied in the
vermicast treatment. b The total amount of sulphate applied with other elements equalizes the sulphate in applied vermicast.
– AES) by extracting sample/solution ratio of 1:25
with Mehlich 3 extraction solution (Mehlich, 1984).
The same extract was used to determine the
extractable phosphorus according to the ammonium
molybdate-ascorbic acid method (Knudsen and
Beegle, 1988). Mineral S in soil was extracted with
0.0125M CaCl2 solution (ratio of soil: solution,
1:4), and analyzed with a turbidimeter after the
addition of BaCl2 and generating BaSO4 turbidity
(Bashour and Sayegh, 2007).
The pH and electrical conductivity (EC) of
the samples were measured in suspension of 1:2
(v/w) by using EI™ 611E EC meter and Digison™
digital pH meter 7007 respectively. Bulk density
was measured on undisturbed cores for soil and the
graduated cylinder method was used for vermicast.
Particle density was determined by volumetric flask
method (Bashour and Sayegh, 2007). The total and
water-filled porosity were calculated from the
particle and bulk density values of respective
samples using standard formulae (Carter and
Gregorich, 2008). Water holding capacity (WHC)
of the samples was obtained by determining their
water retention ability after they were immersed in
water and the excess water was drained off
(Margesin and Schinner, 2005).
2.4. Statistical analysis
One-way ANOVA and post hoc LSD tests
(Weinberg and Abramowitz, 2008) were employed
for determining significance of difference between
the results (SPSS version 16; Softonic, Barcelona,
Spain).
3. Results and discussion
3.1. Seed germination
The GV and GP were significantly higher in
the VC and IF treatments compared to the control
(F2,32 = 3.759, p <0.05; Table 3). The VC
application at the dose of 5 t ha-1
increased the GP
by 8.3% compared to the control. Further increase
in the dose of VC caused an increase in GP to
51.5% higher than the control. In the IF treatments
too, the GP significantly improved with increasing
86
Table 3. Germination value (GV) and germination percentage (GP) of the seeds of cluster bean as influenced
by lantana vermicast and inorganic fertilizers. Results which do not differ significantly (LSD test; p <0.05)
carry at least one character in the superscript which is common.
Treatment Amount Fourth day Fifth day Sixth day Seventh day Eighth day
GV GP GV GP GV GP GV GP GV GP
Vermicompost
5t ha-1 a
3.27 22.86 2.64 25.71 5.10 42.86 4.54 47.14 4.13 51.43
7.5 t ha-1 bc
37.19 77.14 23.80 77.14 17.78 80.00 14.01 82.86 10.73 82.86
10 t ha-1 b
55.57 94.29 37.74 97.14 26.21 97.14 19.26 97.14 14.74 97.14
Inorganic
fertilizer
equivalent to
the VC
5t ha-1 acd
26.99 65.71 17.27 65.71 13.06 68.57 10.41 71.43 7.97 71.43
7.5 t ha-1 bd
34.49 74.29 23.80 77.14 19.07 82.86 14.01 82.86 10.73 82.86
10 t ha-1 b
37.19 77.14 25.60 80.00 19.07 82.86 14.01 82.86 11.48 85.71
Control Nil a 2.50 20.00 2.64 25.71 2.74 31.43 4.26 45.71 3.47 47.14
dosage, and up to 45% higher GP than in control
was recorded at the dose of 10 t ha-1
. Except VC
treatment at 5 t ha-1
, in all other treatments,
maximum GV was observed on the 4th day
followed by a steady decline. In 5 t ha-1
VC, the
maximum GV was recorded on the 6th day. All-in-
all, the maximum GV of 55.6 was recorded with
VC treatment at 10 t ha-1
, followed by 10 t ha-1
of
IF and 7.5 t ha-1
of VC treatments, respectively,
which both had a GV of 37.2.
It is possible that the GP increased with
increased doses of VC and IF as a result of
increasing concentration of nitrate and ammonium
contained in them. As these compounds break
dormancy and stimulate germination (Egley and
Duke, 1985; Hilhorst and Karssen, 2000),
increasing the concentration of VC and IF would
have increased the germination success. In general,
germination is stimulated within a range of 0–0.05
M nitrate and to a certain extent, ammonium ion
also influences the seed germination (Hilhorst and
Karssen, 2000). In the present study, 0.9 to 1.8 g of
nitrate was supplemented in the form of either VC
or IF, which would have increased the germination
in the initial days. Since the nitrate readily leaches
from surface soil by irrigation water, the
concentration of nitrate might have declined as the
days progressed, causing a reduction in germination
success. Moreover, since greater germination
success was recorded with VC treatment than
equivalent IF treatments, phytohormones like
gibberellins and other organic chemicals which are
known to enhance seed germination might have
been involved (Miransari and Smith, 2014) to make
VC more effective than IF.
3.2. Plant growth
VC was seen to exert much more favorable
impact on plant growth than IF, as reflected in all
the plant growth parameters that have been studied
(Table 4). The plants which were treated with VC
showed higher stem diameter, length, fresh
weight/dry weight of shoot, and number of leaves
than plants treated with equivalent IF, or the
control. The beneficial impact of VC may be due to
a number of reasons, including favorable nodules
formation, nutrient release synchronized with plant
need, and the contribution of beneficial plant
growth regulators, hormones and microbes. More
nodules were observed with the plants which were
treated with VC than with IF, and greater
nodulation was witnessed at higher dosage of VC.
This finding is highly significant because
nodulation is a process which is tightly controlled
in nature to allow only as much symbiont rhizobia
in the roots of a legume as necessary for the legume
(Mortier et al., 2012). Several feedback routes exist
in nature to accomplish this control (Miwa et al.,
87
Table 4. Plant growth, leaf pigments, flowering, disease incidence, and retarded growth /death in cluster bean plants as impacted by lantana
vermicast or inorganic fertilizers. Results which do not differ significantly (LSD test; p <0.05) carry at least one character in the superscript which is
common.
*p<0.05,
**p<0.01,
***p<0.001,
n.s - not significant.
Parameters
Vermicompost at dose Inorganic fertilizers at dose
Control
F-value
5 t ha-1
7.5 t ha-1
10 t ha-1
5 t ha-1
7.5 t ha-1
10 t ha-1
Type of
fertilizer Amount
Plant growth
Stem diameter (mm) 13.6±0.7a 14.9±1.2
b 17.2±1.2
c 12.9±1.0
a 13.8±1.4
a 17.0±1.5
c 11.2±1.7
d 18.45
*** 63.69
***
Shoot length (cm) 168±7.0a 203.0±12.6
b 269.7±24.4
c 175.8±12.1
a 195.1±11.8
b 215.4±11.7
d 76.8±5.9
e 80.46
*** 189.0
***
Root length (cm) 74±9.3a 68.2±8.7
ab 65.7±6.9
bc 68.4±5.5
acd 64.9±7.2
bd 52.7±4.5
e 50.2±8.4
e 22.09
*** 20.56
***
Number of leaves 83.6±4.6a 96±5.8
b 99.4±7.2
b 79.3±5.2
ac 81.9±5.3
a 86.5±6.4
ad 42.0±5.3
e 211.2
*** 128.2
***
Number of nodules 62.2±11.3a 69.6±15.2
a 70.7±16.6
a 39.1±7.4
b 35.9±9.9
b 36.1±10.5
b 13.5±6.0
c 101.5
*** 16.03
***
Shoot dry weight (g) 54.2±6.9a 69.3±7.8
b 108.5±13.1
c 19.8±4.7
d 71.5±9.5
b 80.7±11.6
e 45.3±5.2
f 9.690
*** 67.34
***
Root dry weight(g) 5.9±1.0ad
6.5±1.2a 7.7±1.1
b 4.5±1.0
c 5.5±0.9
de 5.1±0.8
ce 3.2±0.6
f 50.18
*** 18.65
***
Leaf pigments
Chlorophyll a (mg g-1
) 2.41±0.06a 3.24±0.13
b 3.79±0.18
c 2.59±0.02
d 2.92±0.05
e 2.95±0.03
e 1.32±0.05
f 92.73
*** 174.4
***
Chlorophyll b (mg g-1
) 2.23±0.06a 2.49±0.07
b 2.88±0.08
c 2.39±0.04
d 2.66±0.06
e 2.72±0.03
f 1.45±0.04
g 143.1
*** 573.7
***
Total chlorophyll (mg g-1
) 4.64±0.07a 5.73±0.13
b 6.68±0.21
c 4.97±0.04
d 5.58±0.07
e 5.66±0.04
b 2.77±0.04
f 111.5
*** 348.5
***
Carotenoids (mg g-1
) 0.49±0.05a 0.57±0.03
b 0.59±0.02
b 0.51±0.02
ac 0.52±0.02
c 0.36±0.02
d 0.26±0.02
e 94.50
*** 47.47
***
Flowering
Number of flowers 7.2±4.7a 14.7±5.0
b 25.4±6.8
c 7.9±3.7
a 16.2±6.7
bd 18.4±8.3
d 0.7±1.0
e 49.96
*** 131.2
***
Disease incident, plant death and stunted plants
Number of infected plant 14a 5
a 9
a 18
a 10
a 11
a 16
a 0.735
n.s 1.228
n.s
Number of plant died Nila Nil
a 2
a Nil
a 1
a 2
b Nil
a 0.402
n.s 2.022
n.s
Number of stunted plant Nila 1
a Nil
a 1
a 12
a 5
a 11
a 1.824
n.s 1.026
n.s
88
2006; Miyazawa et al., 2010). Evidently, VC
generates signals that favour nodule formation. In
contrast IF treatments may induce depression in
nodulation because they provide the nitrogen
contained in the IF all at once leading to signals
that discourage nodulation (Carroll and Gresshoff,
1983; Harper and Gibson, 1984; Nie, 1989). van
Schreven (1959) had reported suppression of
growth in clover (Trifolium repens) and lucerne
(Medicago sativa) nodules when urea was applied
at concentration as low as 0.5%. Presence of higher
Mn in the IF may also have contributed to the
suppression of nodulation in the IF treatment
(Dobereiner, 1966; Foy et al., 1978). Even though
VC also contains the same inorganic nutrients, and
in the same concentration, as IF, the release from
VC might have been at much slower rate because of
the predominantly organic matrix of the VC. This
might have made nutrients available to the legume
at a rate best suited to promote nodules formation.
The slow release of nutrients by the VC may
also have caused lesser nutrient loss from the
rhizosphere. The presence of biologically active
substances such as fulvic acids, humic acids and
phytohormones in the VC may have contributed to
better plant growth in VC treated plants.
Specifically, cytokinins have been reported in
vermicast (Zhang et al., 2014). It has also been
shown that humic acids derived from the vermicast
induce morphogenetic and biological changes
favorable to plants which are similar to those
produced by indole-3-acetic acid (Muscolo et al.,
1999). These bioactive substances are probably
produced due to the abundance of microbial
communities in the vermicompost, specifically the
actinomycetes and fungal species, which then
releases phytohormones in the soil (Frankenberger
and Arshad, 1995). Improvement in the physical
properties of the soil, such as aeration and water
holding capacity, may have also contributed to an
increase in the plant productivity in soil treated
with VC (Edwards, 2004).
3.3. Photosynthetic pigments
Photosynthetic pigments chlorophyll and
carotenoid in the leaves of plants were significantly
influenced by the VC and IF application at different
doses (Table 4). The pigment levels were seen to
increase with increasing rate of fertilizer
application as there was concomitant increase in the
nutrient availability (Mengel and Kirkby, 1987;
Shadchina and Dmitrieva, 1995; Ruza, 1996;
Tejada et al., 2007). Higher concentrations of the
pigments were recorded in the VC treatments of 7.5
and 10 t ha-1
than in the equivalent IF treatments.
Only in VC treatment at 5 t ha-1
lesser amount of
chlorophyll a and b was found than in the
equivalent inorganic fertilizer treatment. The
carotenoid content was also higher in the VC
treated plants than the IF treated ones. This also
confirms that VC application increased the nutrient
availability in soil and enhanced nutrient uptake by
plants in comparison to the IF treatment.
3.4. Flowering
Plants grown in soil treated with VC had
significantly larger number of flowers than plants
grown in IF- treated soil (p<0.001; Table 4).
Greater the fertilizer application, more profuse was
the flowering; the trend with both VC and IF being
10 t ha-1
> 7.5 t ha-1
> 5 t ha-1
> control.
3.5. Disease incidence, plant death and stunted
growth
During the first two months of the
experiments, the plants were found to be infected
with bacterial blight (Xanthomonas campestris),
and alternaria blight (Alternaria spp) and infested
with whiteflies (Bemisia tabaci) (Table 4). The
maximum infestation was seen in the 5 t ha-1
IF
treatments followed by control and 5 t ha-1
VC
treatments. Lesser number of infected plants were
observed in VC fertilized plants than in the plants
that were given equivalent IF treatments. Previous
89
Table 5. Harvest index and yield attributes of plants as impacted by lantana vermicast and equivalent
inorganic fertilizers (mean ±SD). Results which do not differ significantly (LSD test; p <0.05) carry at least
one character in the superscript which is common.
*p<0.05, **p<0.01, ***p<0.001,n.s - not significant.
studies on vermicast obtained from animal manure
have shown that the vermicast possesses pest–
repellant properties (Edwards et al., 2011). The
present studies indicate that the lantana vermicast
may also be imbibed with a similar attribute. The
better nutrient availability made possible by the VC
might also be contributing to the development of
resistance to pathogens in the plants because
availability of adequate nutrients enhances the
ability of the plants to limit the penetration,
development, and/or reproduction of invading
pathogens (Graham and Webb, 1991). Some of the
nutrients are also involved in the production of
antimicrobial compounds such as flavonoids and
phenolics that act against the plant pathogens
(Graham and Webb, 1991; Hill et al., 1999). The
humic acid content of the vermicast is also likely to
affect biochemical processes in the plants and
bacteria, resulting in induction of resistance in
plants to certain phytopathogens (Sahni et al.,
2008).
Moreover, lesser number of VC treated plants
exhibited stunted growth than the IF treated ones
and the control (Table 4). A few plants died during
the experiment, but there was no significant
difference in the number among different
treatments (Table 4).
3.5. Fruit yield
The number and weight of pods per plant
were significantly higher in VC treatments than the
equivalent IF treatments (p<0.05) (Table 5).
Among all the treatments, highest yield of pods was
recorded with VC treatment at 7.5 t ha-1
followed
by the equivalent IF treatment. The weight and the
number of fruits harvested from 7.5 t ha-1
VC
treated plants was about one fold greater than the
corresponding IF treatment. At higher fertilizer
dose (10 t ha-1
) this beneficial effect was reversed.
It is known that application of nutrients which is
well above the levels required by a plant may
induce higher immobilization of nutrients in the
plant biomass and less partitioning to fruits (Wada
et al., 1989; Sujatha and Bhat, 2013). A similar
factor might have been behind the reduction of
yield in the 10 t ha-1
VC/IF applications.
Table 5 shows that the harvest index (HI)
values of VC treated plants are not significantly
higher than that of the IF treated plants even as the
former have given significantly higher fruit yield.
This is because HI is a ratio of mass of fruits and
mass of the above-ground biomass. In VC treated
plants both are higher than in IF treated plants
leading to similar HI.
Parameters, average
value
Vermicompost at dose Inorganic fertilizer at dose F-value
5 t ha-1 7.5 t ha-1 10 t ha-1 5 t ha-1 7.5 t ha-1 10 t ha-1 Type of
fertilizer Amount
No.of pod per plant 18.7±6.6a 40.0±8.2c 29.9±7.2b 14.1±5.9d 25.5±6.3b 20.5±7.1a 119.1*** 95.50***
Weight of the pods (g) 93.5±29.0a 227.0±45.0b 162.5±39.5c 77.9±31.9d 145.4±37.4e 110.5±36.5a 139.0*** 154.8***
Pod length (cm) 15.1±1.7a 16.0±1.4b 16.0±1.6b 14.8±1.8a 15.0±1.8a 14.7±1.8a 18.63*** 2.744n.s
Pod width (cm) 10.2±1.6a 11.3±1.5b 10.6±0.8ac 10.6±1.4ad 11.0±1.4bcd 10.8±1.9bcd 0.385n.s 6.249**
Pod thickness (cm) 6.19±0.84aeg 6.66±0.79b 6.35±0.5ab 5.87±1.02adg 6.02±0.71cdef 5.87±0.62fg 24.18*** 3.471*
No.of seeds per pod 9.20±1.25ab 9.56±0.97a 9.42±1.20ac 9.02±0.94bc 9.48±1.09a 9.46±0.95a 0.368n.s 4.116*
Seed diameter (mm) 5.60±0.86a 6.14±0.72b 6.47±0.73c 5.59±0.97a 6.00±0.76d 6.52±0.71c 0.917n.s 294.7***
Seed thickness (mm) 2.24±0.47a 2.49±0.45b 2.54±0.55bc 2.07±0.63d 2.37±0.55e 2.55±0.47c 17.13*** 132.8***
Harvest Index % 69.8±40.1abcde 107.0±71.2be 50.4±37.1c 74.8±50.8abcde 91.9±78.8ab 51.3±34.8cde 0.965n.s 5.107*
90
The length, width and thickness of the pods,
and the average number of seeds per pod, were
higher in plants treated with VC than in the
equivalent IF treatment. The VC and IF treatments
at 7.5 t ha-1
showed these yield attributes in better
measure than the 5 and 10 t ha-1
treatments,
whereas seed diameter and seed thickness were
maximum in 10 t ha-1
treatments.
3.6. The present study in the context of the state-
of-the-art
There is only one pre-existing report on this
subject, that of Suthar and Sharma (2013). They
had made four blends of lantana and cow manure
(CM) in 1:4, 1:1.5, 1.5:1, and 4:1 mass ratios,
composted it for 3 weeks, then kept 250 g dry
weight equivalent of each pre-composted blend
with 10 individuals of Eisenia fetida for 60 days.
They then found that the resulting worm-worked
reactor content supported germination of the seeds
of corn (Zea mays). It is difficult to conclude much
from this study because each treatment had so
much CM in it that the very low number of
earthworms, 10, that were used in each treatment
might as well have ingested CM particles for most
part. Secondly, once lantana or any other
phytomass is brought in contact with CM, the
cellulolytic and acidogenic bacteria contained in the
CM begin biodegrading the phytomass to produce
volatile fatty acids (VFAs; Kumar et al., 2014).
Suthar and Sharma (2013) have reported that there
was a sharp fall in pH in their treatments which also
indicates that VFA formation might have occurred.
This is likely to have effected the entire chemistry
and microbiology of the treatments and much of the
data reported by Suthar and Sharma (2013) is likely
to have been influenced by this happening. In
particular, lowering of pH is known to make trace
nutrients more labile which might have contributed,
rather than vermicomposting, in the beneficial
effects on the germination of corn as seen by them.
Moreover, unless vermicomposting is defined and
quantified in terms of vermicast deposited by the
earthworm―which is always an easily
distinguishable, separable, and quantifiable entity in
any vermireactor (Tauseef et al., 2014) ― it is
never possible to design and optimize a
vermireactor. The worm–worked substrate, as
deemed vermicompost by Suthar and Sharma
(2013) and other authors before them (Nayeem-
Shah 2014), is a non-descript entity, being a
mixture of vermicast, partially biodegraded
substrate, and numerous products of natural and
worm-mediated biodegradation occurring
uncontrolled in such vermireactors, of which exact
proportions or even nature is very difficult to
quantify. Nor based on such a parameter, it can be
said when the vermireactor attains steady state or
when the vermicomposting is complete. As
vermicast is not periodically removed from these
reactors, but is instead periodically homogenized
with the rest of the reactor content, the earthworm
may be reingesting much of it, thereby harming the
reactor efficiency while the uncontrolled
biodegradation of the rest of the unharvested
vermicast would make process monitoring or
control even more difficult.
Due to all these reasons, it is difficult to draw
any meaningful conclusion from the study of Suthar
and Sharma (2013) on either vermicomposting of
lantana or its fertilizer value. In contrast, the
present report is based on lantana vermicompost
which was generated by direct vermicomposting of
lantana with Eudrilus eugeniae in pulse-fed, high-
rate, vermireactors by a process detailed elsewhere
(Kumar et al., 2012). The vermicast, which was
periodically harvested in each pulse, was obtained
as a clear and precisely quantifiable product of the
vermireactors and was termed vermicompost. The
scientific basis of this definition has been given
earlier (Abbasi et al., 2009), and has been included
as a valid definition in Edward et al. (2011). In
view of this it can be said that the present report
shows rather conclusively that vermicompost
derived from phytomass can be as plant-friendly as
manure-based vermicompost is known to be, even
when it is derived from an allelopathic weed like
91
lantana which, additionally, possesses mammalian
toxicity.
4. Conclusions
The paper describes results of a study, the
first of its kind, in which the effect of
vermicompost (VC) derived solely from an
allelopathic weed has been studied on the
germination, growth and fruition of a botanical
species. The impact of the VC was compared with
that of an inorganic fertilizer (IF) which had all the
main macro and micro – nutrients in concentrations
equivalent to the ones present in the VC. Several
sets of experiments were carried out in which the
soil was treated with either VC obtained from
lantana (Lantana camara) or the IF at the rates of 5,
7.5 or 10 tons per hectare (t ha-1
). Cluster bean
(Cyamopsis tetragonoloba) was the botanical
species used in the study; chosen because it is a
common vegetable and a legume. Additionally a set
of controls was studied in which the soil was used
without fortification by either VC or IF.
It was seen that significantly greater
germination rates occurred in the VC treatments
compared to controls. VC also supported better
plant growth in terms of stem diameter, shoot
length, shoot mass, number of leaves, and leaf
pigments. The positive impact extended up to pod
yield. In addition, vermicast application enhanced
root nodule formation, reduced disease incidence,
and permitted lesser number of stunted plants.
The findings reveal that the allelopathic
ingredients of lantana seem to have been totally
eliminated during the course of its
vermicomposting, and that lantana vermicompost
has the potential to support germination, growth
and fruit yield better than equivalent quantity of
inorganic fertilizers. The study opens up the
possibility that other allelopathic weeds, as also
plants which are toxic in other ways, may be
utilizable as substrates in high-rate vermireactors as
vermicomposting is likely to destroy the toxic
components of these substrates as it is seen to have
done in case of lantana. This, in turn, may
enormously enhance the applicability of
vermicomposting as well as provide a means of
utilizing the biomass of several invasives which,
otherwise, goes to waste.
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96
Supplementary material
Table 4A. Stem diameter (mm) of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
1.1 1.4 2.1 3.4 4.2 4.9 7.6 8.2 10.3 11.2 13.1 13.6
7.5 t ha-1
1.6 1.7 2.4 3.5 4.6 8.1 8.2 8.4 11.1 12.5 13.2 14.9
10 t ha-1
1.7 2.0 2.2 4.1 6.6 10.0 10.7 11.5 13.6 14.8 15.5 17.2
Inorganic
fertilizer
5 t ha-1
1.3 1.9 2.2 3.7 4.6 5.2 6.2 7.3 8.7 9.8 11.9 12.9
7.5 t ha-1
1.6 2.0 2.1 2.5 4.6 7.3 7.6 8.9 9.4 12.2 12.4 13.8
10 t ha-1
1.7 2.0 2.1 3.2 4.7 6.7 8.3 8.8 10.2 12.4 13.4 17.0
Control Nil 0.7 1.8 2.0 2.3 4.5 5.2 5.8 6.1 1.2 10.8 11.1 11.2
Table 4B. Shoot length (cm) of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
9.8 13.4 15.6 17.1 27.8 42.2 68.4 75.7 91.2 105 115 168
7.5 t ha-1
9.9 11.5 16.1 20.5 36.5 54.9 74.5 106.0 115.8 138 166 203
10 t ha-1
11.5 13.5 18.9 21.0 42.8 62.2 111.5 129.5 148.0 194 220 270
Inorganic
fertilizer
5 t ha-1
10.1 14.1 17.5 18.9 28.4 45.8 70.6 78.2 95.7 117 129 176
7.5 t ha-1
10.5 12.3 18.7 19.9 29.7 49.2 76.7 97.0 101.3 119 152 195
10 t ha-1
10.9 13.3 18.4 20.1 30.0 52.5 83.2 103.5 105.5 129 148 215
Control Nil 3.5 11.4 11.8 12.5 18.0 21.8 23.0 26.0 34.5 42.0 59.3 76.8
Table 4C. Root length (cm) of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
6.3 13.9 22.0 26.9 28.5 38.6 52.5 63.5 65.4 70.2 72.1 74.0
7.5 t ha-1
6.7 19.4 25.3 21.0 26.6 37.6 49.6 62.0 63.4 64.0 66.6 68.2
10 t ha-1
8.9 17.1 20.0 20.5 24.5 37.0 44.0 48.7 56.3 62.1 63.2 65.7
Inorganic
fertilizer
5 t ha-1
6.7 19.4 21.0 28.5 29.3 31.6 40.5 58.7 59.2 61.7 63.6 68.4
7.5 t ha-1
8.2 19.8 20.1 32.4 27.9 29.6 38.5 56.9 58,3 60.2 61.3 64.9
10 t ha-1
8.7 20.5 20.8 21.0 25.2 28.3 37.9 47.6 51.3 53.9 56.4 52.7
Control Nil 3.8 13.7 14.0 23.1 29.1 29.2 37.4 42.0 45.7 47.5 49.3 50.2
97
Table 4D. Number of leaves of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
03 03 04 9 18 28 34 42 60 69 72 84
7.5 t ha-1
03 04 06 12 22 36 45 52 58 70 78 96
10 t ha-1
03 04 06 13 22 37 49 55 63 81 88 99
Inorganic
fertilizer
5 t ha-1
03 03 04 10 19 26 32 38 58 59 63 79
7.5 t ha-1
03 04 07 11 20 30 34 46 62 63 72 82
10 t ha-1
03 04 08 12 21 32 38 50 64 68 72 87
Control Nil 03 03 04 09 15 17 21 25 28 31 35 42
Table 4E. Number of nodules of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
Nil 11 12 15 11 22 34 38 33 52 60 62
7.5 t ha-1
Nil 14 15 24 18 32 40 48 60 52 66 70
10 t ha-1
Nil 15 14 15 22 27 36 65 45 115 82 71
Inorganic
fertilizer
5 t ha-1
Nil 10 14 12 15 6 37 25 12 19 31 39
7.5 t ha-1
Nil 3 7 6 10 22 12 46 26 47 17 36
10 t ha-1
6 17 16 10 14 24 16 38 41 22 30 36
Control Nil Nil 0 7 8 12 14 19 11 11 9 16 14
Table 4F. Shoot dry weight (g) of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.033 0.210 0.295 0.551 0.967 4.914 7.990 13.80 25.14 36.25 48.29 54.17
7.5 t ha-1
0.076 0.310 0.486 0.738 2.971 7.312 12.30 20.13 34.72 52.05 60.48 69.27
10 t ha-1
0.090 0.346 0.601 1.080 4.252 11.36 28.50 35.14 54.63 68.53 87.82 108.5
Inorganic
fertilizer
5 t ha-1
0.055 0.211 0.354 0.359 1.169 3.871 6.184 11.30 15.82 23.61 27.56 19.82
7.5 t ha-1
0.128 0.247 0.322 0.429 1.359 4.570 10.11 14.74 21.86 41.71 47.49 71.54
10 t ha-1
0.102 0.304 0.464 0.470 1.465 6.784 16.58 21.50 29.27 46.74 60.48 80.66
Control Nil 0.032 0.109 0.205 0.395 0.955 1.490 4.572 6.799 12.11 41.50 44.08 45.32
98
Table 4G. Root dry weight (g) of cluster bean at different weeks
Table 4H. Leaf chlorophyll a (mg g-1
) content of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.359 1.475 1.521 1.899 1.059 2.131 2.197 2.238 2.297 2.316 2.395 2.407
7.5 t ha-1
0.305 1.477 1.734 1.967 2.291 2.486 2.781 2.931 3.062 3.155 3.197 3.240
10 t ha-1
0.563 1.954 2.192 2.802 2.826 3.169 3.447 3.501 3.587 3.626 3.684 3.793
Inorganic
fertilizer
5 t ha-1
0.581 1.24 1.597 1.88 2.069 2.243 2.331 2.402 2.436 2.491 2.538 2.586
7.5 t ha-1
0.491 1.604 1.934 2.159 2.214 2.356 2.555 2.725 2.762 2.834 2.909 2.916
10 t ha-1
0.323 1.181 1.925 2.175 2.253 2.44 2.755 2.792 2.852 2.873 2.922 2.945
Control Nil 0.103 0.465 0.64 0.907 1.128 1.164 1.177 1.193 1.201 1.224 1.238 1.316
Table 4I Leaf chlorophyll b (mg g-1
) content of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.206 0.568 1.173 1.258 1.306 1.584 1.789 2.058 2.126 2.170 2.201 2.230
7.5 t ha-1
0.262 0.603 0.874 1.151 1.363 1.591 1.822 2.316 2.394 2.446 2.482 2.491
10 t ha-1
0.241 0.587 1.271 1.421 1.782 1.975 2.395 2.550 2.673 2.719 2.783 2.884
Inorganic
fertilizer
5 t ha-1
0.193 0.565 0.938 0.976 1.129 1.779 1.947 2.198 2.228 2.297 2.342 2.385
7.5 t ha-1
0.261 0.511 1.212 1.413 1.747 1.867 2.193 2.435 2.512 2.579 2.614 2.663
10 t ha-1
0.358 0.552 1.249 1.593 1.812 1.909 2.246 2.466 2.511 2.579 2.652 2.719
Control Nil 0.032 0.155 0.223 0.504 1.612 1.935 1.065 1.409 1.510 1.532 1.544 1.451
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.013 0.021 0.051 0.071 0.184 0.514 0.680 2.403 2.922 3.829 4.933 5.925
7.5 t ha-1
0.010 0.022 0.059 0.087 0.343 0.761 0.982 2.782 3.360 4.259 5.343 6.511
10 t ha-1
0.013 0.028 0.106 0.113 0.421 0.901 1.200 2.913 3.531 4.680 5.689 7.680
Inorganic
fertilizer
5 t ha-1
0.009 0.031 0.036 0.054 0.179 0.435 0.520 1.771 2.660 3.736 4.472 4.532
7.5 t ha-1
0.011 0.044 0.054 0.068 0.202 0.530 0.832 2.316 3.083 4.143 4.905 5.539
10 t ha-1
0.009 0.053 0.083 0.105 0.347 0.658 0.999 2.585 3.397 3.984 5.000 5.121
Control Nil 0.001 0.014 0.016 0.029 0.102 0.256 0.289 0.981 1.763 2.929 3.160 3.214
99
Table 4J. Leaf total chlorophyll (mg g-1
) content of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.565 2.043 2.694 3.157 2.365 3.715 3.986 4.296 4.423 4.486 4.596 4.637
7.5 t ha-1
0.567 2.08 2.608 3.118 3.654 4.077 4.603 5.247 5.456 5.601 5.679 5.731
10 t ha-1
0.804 2.541 3.463 4.223 4.608 5.144 5.842 6.051 6.260 6.345 6.467 6.677
Inorganic
fertilizer
5 t ha-1
0.774 1.805 2.535 2.856 3.198 4.022 4.278 4.600 4.664 4.788 4.880 4.971
7.5 t ha-1
0.752 2.115 3.146 3.572 3.961 4.223 4.748 5.16 5.274 5.413 5.523 5.579
10 t ha-1
0.681 1.733 3.174 3.768 4.065 4.349 5.001 5.258 5.363 5.452 5.574 5.664
Control Nil 0.135 0.62 0.863 1.411 2.740 3.099 2.242 2.602 2.711 2.756 2.782 2.767
Table 4K. Leaf carotenoids (mg g-1
) content of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.138 0.179 0.215 0.253 0.299 0.311 0.32 0.376 0.384 0.414 0.426 0.490
7.5 t ha-1
0.151 0.276 0.352 0.381 0.416 0.428 0.466 0.481 0.499 0.512 0.535 0.572
10 t ha-1
0.116 0.285 0.394 0.421 0.441 0.467 0.475 0.493 0.506 0.538 0.562 0.589
Inorganic
fertilizer
5 t ha-1
0.125 0.206 0.319 0.358 0.395 0.418 0.434 0.457 0.465 0.482 0.491 0.511
7.5 t ha-1
0.154 0.255 0.335 0.379 0.407 0.417 0.427 0.456 0.471 0.489 0.506 0.524
10 t ha-1
0.151 0.183 0.214 0.237 0.252 0.285 0.312 0.314 0.318 0.337 0.341 0.359
Control Nil 0.102 0.122 0.136 0.158 0.164 0.191 0.203 0.218 0.224 0.236 0.248 0.261
100
EFFECT OF VERMICAST GENERATED FROM A
PERNICIOUS WEED IPOMOEA (IPOMOEA CARNEA)
ON SEED GERMINATION, PLANT GROWTH, AND
YIELD OF CLUSTER BEAN (CYAMOPSIS
TETRAGONOLOBA)
Chapter
7
101
A paper based on this chapter has been
submitted for publication.
CChhaapptteerr 77
Effect of vermicast generated from a pernicious weed ipomoea
(Ipomoea carnea) on seed germination, plant growth, and yield
of cluster bean (Cyamopsis tetragonoloba)
Abstract
The impact of vermicast derived from the highly pernicious, amphibian weed, Ipomoea carnea on cluster
bean (Cyamopsis tetragonoloba) was assessed in terms of germination, growth and fruit yield. Seeds of
cluster bean were sown in soils to which vermicast was applied at the rates of 5, 7.5 and 10 t ha-1
. The impact
of vermicast on seed germination, growth and yield of experimental plants were compared with the plants
which were treated with inorganic fertilizers (IF) in concentrations equivalent to those present in the
respective vermicompost (VC) treatments. Additionally a set of control was studied in which the soil was
used without fortification by either vermicompost or inorganic fertilizers. Following assessment of
germination rates, the plant growth parameters were determined each week for 3 months with randomly
collected samples. The yield attributes were assessed with manually harvested mature pods from all the
experimental plants. The effect of different treatments on germination, growth and yield attributes are very
significant. The vermicast treated plants showed better growth and yield compared to the respective
inorganic fertilizer treatment and control. However, there was suppressive effect on germination with
vermicast treatment.
1. Introduction
Ipomoea carnea (family: Convolvulaceae) is
an amphibious toxic weed widely distributed in
India (Konwer et al., 2007; Abbasi and Abbasi,
2010a) and in other tropical and subtropical
countries (Austin and Huaman, 1996). This plant is
native to South America, and it was introduced into
India as an ornamental plant at the end of the 18th
century (Haines, 1925). Since then it has spread
rapidly in the many parts of the country. This weed
colonizes large tracts of water-bodies and land
areas and causing severe damages to the native
vegetation and fauna. In addition to this, the dead
biomass generated from this and other weeds
deposited at benthic zone of water reservoirs, and
undergo anaerobic decomposition which generates
enormous quantity of methane (Abbasi and Abbasi,
2010b). Methane has 25 times the greenhouse
impact of CO2 and so its release to the atmosphere
is of concern (Abbasi and Abbasi, 2010b, 2012).
At present very few effective weed
management tools are available for control (Abbasi
and Abbasi, 2010a). Chemical herbicides enable
control of these weeds quickly and efficiently, but
temporarily. There is also an environmental cost in
using herbicides. Biological control can be an
environmentally safe but selection and maintaining
of host-specific natural enemies makes the process
complex and challenging. The prolific growth and
rapid regeneration of the weed even after cutting,
also defies physical means of destruction and
control (Ganesh et al., 2008). Although, the
102
ipomoea generates huge biomass, it cannot be used
as firewood due to its poor heating value and as it
emits poisonous gases during the burning process
(Konwer et al., 2007). In some part of the country,
this weed is being used for rural housing, and
fencing in agricultural lands. This weed is also used
as laxative, and reported to be traditional healer for
leucoderma and other skin related diseases, and to
provoke menstruation (Jain et al., 2009). However,
only very small quantity of this weed is utilizable
for these purposes and also in very limited
situation. The huge quantity of this weed biomass
can be utilized for generating organic fertilizer such
as vermicompost and compost. This may provide
an answer to the minimization of this weed biomass
accumulation. In addition to this, application of
vermicompost/compost reduces the widespread
deterioration caused to agricultural land due to
rampant use of inorganic fertilizers. Since huge
amount of energy is required for fertilizer
manufacturing and, there is release of greenhouse
gases and other toxic and hazardous wastes at
chemical fertilizer factories, adopting to the organic
farming techniques will be the economically and
environmentally feasible alternative for chemical
fertilizers.
Studies by several authors (Gajalakshmi et
al., 2001, 2002, 2005; Yadav and Garg, 2011;
Makhija et al., 2011) have shown that by
vermicomposting process, most of the weeds can be
converted to vermicast. But it has to be ascertained
whether these vermicast are beneficial to plants.
Such an assessment is particularly necessary for the
weed like ipomoea as it is known to contain
allelopathic compounds (Patel et al., 2009).
Therefore, attempt has been made in this report to
assess the effect of vermicompost generated from
this weed on the germination and growth of cluster
beans (Cyamopsis tetragonoloba). The response of
this plant to vermicompost generated from ipomoea
has been compared with equivalent inorganic
fertilizer treatments.
2. Materials and methods
The experiments were conducted at
Pondicherry University, Puducherry, India
(11°56’N, 79°53’E). This region located on the east
coast of Indian peninsula has a typical maritime
tropical climate with a dissymmetric rainfall. The
mean annual rain fall of this region is about 1300
mm with 57.25 mean rainy days, restricted mainly
during October to December. The experiments were
conducted in 49 liter volume wooden containers (40
cm height with surface area 35 x 35 cm), lined with
high-density polyethylene (HDPE) sheets. The
barren land soil was used in the experiments to
minimize the errors due to the earlier soil practices.
The experimental soil was characterized as sandy
loam soil with low organic carbon and nutrients
content. The leaves of Ipomoea carnea
vermicomposted by an epigeic species, Eudrilus
eugeniae by using direct vermicomposting process
(Makhija et al, 2011), as detailed in chapter 5. The
physico-chemical properties of the vermicast and of
the soil with which it was used are given in Table 1.
The experiments were carried out during July –
October which is ideal for growing cluster beans in
the study area (ICAR, 2011).
For studying the impact of vermicast on
cluster bean, three sets of experiments were
conducted. Based on the nitrogen, phosphorus, and
potassium (NPK) concentrations seen in the
vermicompost (Table 1), and the levels of NPK
needed in the soil for cluster bean (ICAR, 2011), it
was calculated that the cluster bean would require
7.5 t ha-1
of ipomoea vermicompost (VC) for its
optimum growth. In the first set, one batch of
containers was supplied with 7.5 t ha-1
of VC while
two other batches were given 5 and 10 t ha-1
of VC.
Second set comprised of chemical fertilizers (IF)
which were treated with nutrients N, P, K, Ca, Mg,
S, Fe, Mn, Cu, Zn, B, Mo and Cl in concentrations
equivalent to those present in the respective
vermicompost treatments (Table 2). The nutrients
were supplied in two installments: half at the time
103
Table 1. Chemical and physical properties of vermicast
and soil used in the study
BDL – Below detection limit
of sowing and the rest at the onset of flowering.
The last set had no supplementation of nutrients
and served as control. In this manner 3 sets of
experiments, encompassing seven treatments were
carried out. As each treatment had 36 containers,
each with one plant, 252 containers were utilized
for the main experiment. Another 150 containers
were maintained with all the treatments, as spares
which were used to replace the containers of which
plants were sacrificed for analysis, or in which the
plants happened to have died. The Pusanavabahar
variety cluster bean, which is locally available, was
used. Following the assessment of germination over
eight days, growth and yield were monitored for an
additional three months. Throughout the
experiments, adequate watering was performed to
maintain 20-30% (v/w) moisture. Deweeding was
done periodically. Neem extract was applied as a
mild pesticide in a few instances when pests were
seen infesting the plants.
2.1. Germination, plant growth and yield
characteristics
Seed germination was assessed for 8 days
from the day of sowing and was quantified in terms
of germination value (GV) using the formula of
Djavanshir and Pourbeik (1976):
Where GV is germination value; DGS is daily
germination speed which is computed by dividing
cumulative germination percent by the number of
days elapsed since the beginning of the test; GP is
germination percentage at the end of the test, and N
is frequency or number of DGS that are calculated
during the test. The unit for GV is germinated
seeds/day. It is used as an index to statistically
assess the effects of different treatments.
The growth was assessed on the basis of stem
diameter, length of shoot and root, number of
leaves, number of nodules present in the root, the
nodules size and shoot/root biomass for 3 months.
Plants were harvested periodically, and at each
harvest, the fruits were counted and their fresh and
dry weights were determined. To quantify biomass
in terms of dry weight, samples were oven-dried at
85°C to constant weight. Chlorophyll a and b, and
carotenoids were extracted by the method described
by Moran and Porath (1980). One gram of mature
leaf was ground and incubated with N,N-di-methyl
formamide (DMF) for 24 h at 4 °C in dark, shaking
every 6 hours (Moran and Porath, 1980). The
resultant supernatant was read at 470, 647 and 664
nm and the concentration of pigments were
Variables Concentration
Vermicast Soil
Chemical properties
pH 6.51±0.06 6.30±0.10
Total organic carbon g kg-1
356.8±12.1 8.87±0.02
Total Nitrogen g kg-1
19.8±0.2 2.66±0.02
Plant available form of
Phosphorus mg kg -1
37.83±0.60 0.41±0.01
Potassium g kg-1
2.12±0.04 0.40±0.00
Sulphur mg 100g-1
4.64±0.09 0.54±0.01
Calcium g kg-1
15.5±2.6 8.27±0.01
Magnesium g kg-1
4.85±0.09 0.09±0.02
Boron mg kg-1
78.1±2.3 26.9±1.2
Copper mg kg-1
10.6±0.3 5.08±0.15
Iron mg kg-1
270.3±14.7 59.9±2.9
Manganese mg kg-1
117.4±5.0 45.1±2.2
Zinc mg kg-1
107.0±7.7 55.0±2.6
Molybdenum mg kg-1
BDL BDL
Physical properties
Dry weight % 48.4±1.1 94.7±0.1
Bulk density g cm-3
0.26±0.02 1.28±0.00
Particle density g cm-3
1.30±0.05 2.70±0.12
Water-holding capacity % 261.9±16.6 36.9±2.4
Electrical conductivity
mmhos cm-1
6.25±0.07 0.12±0.02
Total porosity % 80.2±1.8 53.1±2.0
Air filled porosity % 70.0±2.6 46.5±2.1
104
Table 2. Amount of inorganic fertilizer applied equivalent to vermicast treatment.
Nutrients Form in which
applied
Mass %
of nutrient
in applied
compound
10t ha-1
7.5t ha-1
5t ha-1
Concentra
tion of
nutrient in
VC
(kg ha-1
)
Amount
of IF
applied
(kg ha-1
)
Concentration
of nutrient in
VC
(kg ha-1
)
Amount
of IF
applied
(kg ha-1
)
Concentration
of nutrient in
VC
(kg ha-1
)
Amount
of IF
applied
(kg ha-1
)
Nitrogen CH4N2O 46.65 68.22 142.7a 51.16 107.0
a 34.11 71.34
a
Phosphorus (NH4)2HPO4 23.45 18.37 78.33 13.78 58.75 9.19 39.16
Potassium KCl 52.44 10.41 19.85 7.808 14.89 5.206 9.926
Sulphur 26.93 0.223 Nilb 0.167 Nil
b 0.111 Nil
b
Calcium CaCO3 40.04 75.14 187.7 56.36 140.7 37.57 93.83
Magnesium MgO 60.30 23.44 38.88 17.58 29.16 11.72 19.44
Boron Na2B4O7.10H2O 11.34 0.378 3.331 0.283 2.498 0.189 1.665
Copper CuSO4.5H2O 25.45 0.051 0.201 0.038 0.151 0.026 0.101
Iron FeSO4.7H2O 20.09 1.316 6.552 0.987 4.914 0.658 3.276
Manganese MnSO4. H2O 32.50 0.568 1.747 0.426 1.310 0.284 0.873
Zinc ZnCl2 47.97 0.517 1.078 0.388 0.809 0.259 0.539 a The total amount of N applied in the form of urea and di-ammonium phosphate is equal to the concentration of N applied in the
vermicast treatment. b The total amount of sulphate applied with other elements equalizes the sulphate in applied vermicast.
determined as detailed by Wellburn (1994). The
yield was calculated in the form of harvest index
(HI) which is the ratio of weight of beans per plant
to the above-ground biomass. The yield as reflected
in pod size, pea size, number of peas per pod and
diseased pods, if any, were recorded at each harvest
with randomly selected 50 fruits.
During the course of the experiment, the
plants were infected with bacterial blight
(Xanthomonas campestris) and alternaria blight
(Alternaria spp). Infestation of white fly (Bemisia
tabaci) also occurred. The bacterial blight infection
was seen in the symptoms of black leaf spots,
necrotic lesions at leaf tips, black streaks on
petioles and stems, split stems, and defoliation
(Mihail and Alcorn, 1985; Ren, 2014). Plants were
considered infected by Alternaria spp. when there
were dark brown lesions on leaves with concentric
zonations demarcated with light brown lines. In
severe infections, several spots merged and gave
the leaves a blighted look, eventually causing
defoliation (Orellana and Simmons, 1966;
Yogendra et al., 1995). The number of plants that
died, either with or without any symptoms of
infection, and stunted plants were also recorded.
2.3. Analytical methods
The analytical methods were the same as
detailed in section 2.3 of chapter 6.
2.4. Statistical analysis
One-way ANOVA and post hoc LSD tests
were employed for determining significance of
difference between the results (SPSS version 16;
Softonic, Barcelona, Spain).
3. Results and discussion
3.1. Seed germination
The germination value (GV) (F2,32 = 8.223, p
<0.01) and germination percentage (GP) (F2,32 =
33.02, p <0.001) were significantly influenced by
different treatments (Table 3). In all the treatments,
maximum GV was observed on 4th day since the
beginning of the experiment and then there was a
105
decreasing trend. In most of the treatments, lowest
GV was reported on the 8th day. In the VC
treatment, increase in the dose of application has
shown reduction in the germination rate (Table 3).
A maximum of 80% of GP was observed with 5 t
ha-1
VC treatment and lowest GP of 62.9% with 10
t ha-1
treatments. The increasing IF application
equivalent to VC treatment showed increasing trend
in GP. The IF treatment showed maximum GP of
78.6% with 10 t ha-1
treatment followed by 74.3%
and 68.6% with 7.5 t ha-1
and 5 t ha-1
treatments
respectively. However, a maximum GV of 31.9 was
in 7.5 t ha-1
treatment on the 4th day.
This result indicates that, seed germination may be
strongly influenced by the applied IF constituent,
particularly nitrate and ammonium they contains.
As these compounds break dormancy and stimulate
germination (Egley and Duke, 1985; Hilhorst and
Karssen, 2000), increasing the concentration of IF
would have increased the germination success.
Their nature of readily leaching from surface soil
by irrigation water may be the reason for higher
germination at initial days followed by steep
inclination till the end. In the case of VC, a
maximum germination success was recorded in 5 t
ha-1
VC. With increasing dose of VC application a
declining trend of seed germination was observed.
However, in all the VC treatment, GV was much
higher than the control. Few studies have reported
similar suppressive effect on germination with
increasing concentration of VC generated from
kitchen waste, paper waste, yard waste and cattle
dung (Roberts et al., 2007; Warman and AngLopez,
2010; Ievinsh, 2011). Several factors have been
postulated by different authors for the suppressive
effect of VC on germination, such as high
concentrations of gallic acid and chlorogenic acid
(Reigosa et al., 1999), auxin and gibberlellin-like
substances in humic and fulvic acids of
vermicompost (Arancon et al., 2006). In the present
study, presence of some of these components in
excess quantity might be the reason of suppressive
effect on seed germination with increasing dose of
VC.
3.2. Plant growth
The VC and IF application showed a
differential impact on all the plant growth (Table
4). The plants which were treated with VC grew
faster than the IF treated plants and control. The
plants treated with VC showed about 15% higher
stem diameter, 17% longer shoot length, 33%
higher dry weight of shoot, and 16% higher number
of leaves than those of equivalent IF. Even though
Table 3. Germination value (GV) and germination percentage (GP) of the seeds of cluster bean as influenced
by ipomoea vermicast and inorganic fertilizers. Results which do not differ significantly (LSD test; p <0.05)
carry at least one character in the superscript which is common.
Treatment Amount Fourth day Fifth day Sixth day Seventh day Eighth day
GV GP GV GP GV GP GV GP GV GP
Vermicompost
5t ha-1 a
24.70 62.86 17.27 65.71 13.06 68.57 13.06 80.00 10.00 80.00
7.5 t ha-1 ac
22.50 60.00 15.81 62.86 11.99 65.71 10.41 71.43 9.30 77.14
10 t ha-1 b
14.74 48.57 13.06 57.14 10.00 60.00 7.35 60.00 6.17 62.86
Inorganic
fertilizer
equivalent to
the VC
5t ha-1 cb
16.53 51.43 13.06 57.14 11.99 65.71 8.81 65.71 7.35 68.57
7.5 t ha-1 a
31.89 71.43 22.08 74.29 15.33 74.29 11.26 74.29 8.62 74.29
10 t ha-1 a
29.39 68.57 20.41 71.43 15.33 74.29 12.14 77.14 9.65 78.57
Control Nil d 2.50 20.00 2.64 25.71 2.74 31.43 4.26 45.71 3.47 47.14
106
Table 4. Plant growth, leaf pigments, flowering, disease incidence, and retarded growth /death in cluster bean plants as impacted by ipomoea
vermicast or inorganic fertilizers. Results which do not differ significantly (LSD test; p <0.05) carry at least one character in the superscript which is
common.
*p<0.05,
**p<0.01,
***p<0.001,
n.s - not significant.
Parameters
Vermicompost at dose Inorganic fertilizers at dose
Control
F-value
5 t ha-1
7.5 t ha-1
10 t ha-1
5 t ha-1
7.5 t ha-1
10 t ha-1
Type of
fertilizer Amount
Plant growth
Stem diameter (mm) 16.9±1.5a 22.6±4.2
b 24.1±2.5
b 16.8±1.4
a 18.4±1.2
a 22.9±2.0
b 11.2±1.7
c 36.70
*** 70.94
***
Shoot length (cm) 144.5±5.0a 197.6±9.8
b 231.6±10.5
c 138.6±5.7
a 162.7±7.3
d 191.4±6.9
b 76.8±5.9
e 70.73
*** 202.3
***
Root length (cm) 55.0±8.0a 41.4±4.1
b 38.3±4.8
b 47.7±5.2
c 47.2±4.3
c 39.6±3.9
b 50.2±8.4
c 2.364
n.s 18.33
***
Number of leaves 98.4±5.5a 120.3±7.7
b 133.8±5.6
c 92.3±3.8
d 108.3±6.0
e 112.2±6.6
e 42.0±5.3
f 162.1
*** 236.9
***
Number of nodules 61.3±10.8a 90.3±15.4
b 129.3±23.2
c 31.9±7.2
d 50.0±11.9
e 37.1±7.3
d 13.5±6.0
f 76.96
*** 15.41
***
Shoot dry weight (g) 51.5±5.6a 89.3±6.6
b 102.6±10.8
c 42.9±4.6
d 59.6±5.2
e 82.8±6.6
f 45.3±5.2
d 18.64
*** 74.71
***
Root dry weight(g) 5.02±0.22a 6.25±0.60
b 11.90±2.14
c 4.60±0.74
a 8.10±1.18
d 6.84±0.93
b 3.21±0.58
f 15.21
*** 40.90
***
Leaf pigments
Chlorophyll a (mg g-1
) 2.17±0.14a 3.21±0.25
b 3.45±0.11
c 1.99±0.09
d 2.69±0.10
e 2.72±0.06
e 1.32±0.05
f 58.35
*** 145.0
***
Chlorophyll b (mg g-1
) 2.21±0.05a 2.30±0.06
b 2.47±0.08
c 2.10±0.14
d 2.28±0.10
ba 2.57±0.08
e 1.45±0.04
f 137.5
*** 370.8
***
Total chlorophyll (mg g-1
) 4.38±0.14a 5.51±0.24
b 5.92±0.16
c 4.09±0.08
d 4.97±0.12
e 5.29±0.10
f 2.77±0.04
g 88.05
*** 329.7
***
Carotenoids (mg g-1
) 0.42±0.04a 0.51±0.05
b 0.55±0.08
b 0.36±0.04
c 0.41±0.05
a 0.53±0.07
b 0.26±0.02
d 38.36
*** 59.70
***
Flowering
Number of flowers 27.4±7.0a 37.6±12.4
b 31.7±11.1
cd 15.7±4.2
e 27.9±8.0
ad 18.1±8.7
ce 0.7±1.0
f 158.0
*** 84.87
***
Disease incident, plant death and stunted plants
Number of infected plant 13a 3
a 8
a 17
a 9
a 15
a 16
a 1.201
n.s 1.193
n.s
Number of plant died Nila 1
a Nil
a 2
a 2
a 1
a Nil
a 2.873
n.s 0.874
n.s
Number of stunted plant Nila 1
a 1
a 1
a 5
a 6
a 11
a 2.380
n.s 1.384
n.s
107
the plants from both the VC and IF treatments
received same amount of all the essential nutrients,
there was a drastic difference in its growth, which
probably due to a number of reasons, including
favorable nodules formation, nutrient release
synchronized with plant need, and the contribution
of beneficial plant growth regulators, hormones and
microbes with VC application.
In this experiment, maximum number of
nodules was observed with the plants which were
treated with VC than IF. The increasing dose of VC
application has increased the number of nodules to
the extent of 89%, in comparison with control. In
the case of 5 and 7.5 t ha-1
VC treatments, about
45% higher number of nodules was recorded in
comparison to the equivalent IF treatments and
those of 10 t ha-1
VC showed 89% higher number
of nodules. The results are indicates that VC
application favored nodule formation, in contrast IF
treatments induce depression in nodulation.
Inhibition of nodulation with IF treatments
probably the reason of its high nitrate content
(Carroll and Gresshoff, 1983; Harper and Gibson,
1984; Nie, 1989). van Schreven (1959) had
reported that increasing concentration of urea
application decrease the weight of nodules; even
the lowest concentration of 0.5% exerted this
suppressing effect on clover plants (Trifolium
repens) and lucerne plants (Medicago sativa).
Presence of higher Mn in the IF may also have
contributed to the suppression of nodulation in the
IF treatment (Dobereiner, 1966; Foy et al., 1978).
Even though VC also contains the same inorganic
nutrients, and in the same concentration, as IF, the
release from VC might have been at much slower
rate because of the predominantly organic matrix of
the VC. This might have made nutrients available
to the legume at a rate best suited to promote
nodules formation. The slow release of nutrients by
the VC may also have caused lesser nutrient loss
from the rhizosphere. In addition, supplement of
biologically active substances such as fulvic acids,
humic acids and phytohormones and improvement
in the physical properties of the soil with VC
application also might have contributed to better
plant growth in VC treated plants, which is
discussed in detail in the chapter 6.
3.3. Photosynthetic pigments
Photosynthetic pigments such as chlorophyll and
carotenoid concentration of the plants were
significanly influnced by the applied VC and IF
(p<0.001;Table 4). In general, VC application was
more effective than the respective inorganic
fetilizer treatment. However, in most of the weeks,
VC treatment at the dose of 5 t ha-1
has shown
lesser amount of chlorophyll a and b than the
equivalent inorganic fertilizer treatments. Lower
rate of nutrient application and their poor
availability due to the slower releasing property of
vermicast might have reduced these pigments
concentration with 5 t ha-1
VC treatment. As the
formation of chlorophyll depends on the adequate
supply of nitrogen and iron, the plants which has
supplied with maximum of these nutrients either in
the form of VC and IF had increased its pigment
content. The photoprotection pigments, carotenoids
also found to be synthesized more in the plants
grown in VC treated soil than equivalent IF
treatment.
3.4. Effect on flowering
The VC treated plants produced more number
of flowers in comparison to the equivalent IF
treated plants (p< 0.001) (Table 4). A maximum
number of flowers was recorded with 7.5 t ha-1
treatment with both VC and IF treatments.
Increasing dose fertilizer application showed the
trend of 7.5 t ha-1
> 10 t ha-1
> 5 t ha-1
> control
with VC and equivalent IF treatment. The plants
grown in the soil treated with 7.5 t ha-1
of both VC
and IF showed early flowering than the other
treatments. Adequate availability of nutrient in this
treatment could be the reason for the higher and
early flowering in this study.
108
3.5. Disease incidence, plant death and stunted
growth
The plants were infected with bacterial blight
(Xanthomonas campestris), and alternaria blight
(Alternaria spp) and infested with whiteflies
(Bemisia tabaci) maximally during the first two
months (Table 4). The incidence of disease was not
significantly differing with VC and IF application.
However, a noticeable difference was observed
with the amount of nutrient applied in both VC and
IF treatments. The both VC and equivalent IF
supplemented at the dose of 5 t ha-1
showed
maximum number of infected plants.
Moreover, a few plants died during the
experiment, but there was no significant difference
in the number among different treatments (Table 4).
The maximum number of stunted plants was seen
in the first 12 days since beginning of the
experiment. After a month, all the plants grew well.
Inorganic treatment exhibited more number of
stunted growth plants in comparison to the VC
treatment. In IF treatment at the dose of 10 t ha-1
, a
maximum of 6 stunted plants was recorded,
whereas, in all VC treatments lesser number of
plants exhibited stunted growth.
3.6. Effect on yield
The observation on yield attributes is given in
Table 5. The weight and number of harvested pods
from the experimental plants were significantly
different with different form and amount of nutrient
applied (p<0.001) (Table 8). The plants treated with
VC at the dose of 5, 7.5 and 10 t ha-1
produced 6,
20 and 24% higher mass of fruits, respectively in
comparison to its equivalent IF treatments. In both
VC and IF treatments, the weight and number of
pods harvested were maximum at 7.5 t ha-1
;
whereas, increase dose to 10 t ha-1
, reduced the fruit
yield. Reduction of yield in the 10 t ha-1
VC/IF
applications probably due to the excess nutrient
applied in the treatments which is well above the
levels required by a plant may induced higher
immobilization of nutrients in the plant biomass
and less partitioned to fruits (Wada et al., 1989;
Sujatha and Bhat, 2013).
The pods harvested from the VC treated
plants were lengthier, wider and contains more
number of seeds than those of IF treatments. The
seed diameter and thickness were also higher in VC
than the equivalent IF treatments. In both VC and
IF treatments at 7.5 t ha-1
showed these yield
Table 5. Harvest index and yield attributes of plants as impacted by ipomoea vermicast and equivalent inorganic
fertilizers (mean ±SD). Results which do not differ significantly (LSD test; p <0.05) carry at least one character in the
superscript which is common.
*p<0.05, **p<0.01, ***p<0.001,n.s - not significant.
Parameters, average
value
Vermicompost at dose Inorganic fertilizer at dose F-value
5 t ha-1 7.5 t ha-1 10 t ha-1 5 t ha-1 7.5 t ha-1 10 t ha-1 Type of
fertilizer Amount
No.of pod per plant 20.1±8.1a 36.3±10.0b 28.3±7.2c 19.5±6.7a 30.9±7.8c 22.4±7.5a 103.4*** 91.39***
Weight of the pods (g) 110.1±43.8a 201.2±55.9b 159.3±42.4c 103.7±35.7a 161.4±42.7cd 120.8±41.2a 134.8*** 147.8***
Pod length (cm) 14.6±1.8a 15.8±1.6b 15.5±1.6b 14.1±1.6a 14.7±1.9a 14.3±1.7a 22.24*** 6.851**
Pod width (cm) 10.2±1.0a 10.8±0.9bd 16.0±1.3c 10.2±1.3ade 10.4±0.9be 10.8±1.5b 53.32*** 83.730***
Pod thickness (cm) 6.39±0.85a 6.01±0.75b 6.79±0.58c 5.80±0.54b 5.99±1.04b 5.74±0.74b 36.21*** 2.604 n.s
No.of seeds per pod 9.24±0.92a 9.50±1.16a 9.50±1.16a 9.18±1.16a 9.30±1.28a 9.18±1.12a 2.175 n.s 0.728 n.s
Seed diameter (mm) 5.59±1.01a 6.01±0.86b 6.23±1.69c 5.61±0.95a 5.96±0.89b 6.51±0.72d 5.565* 170.9***
Seed thickness (mm) 2.30±0.57a 1.90±0.59b 2.43±0.59c 2.14±0.63d 2.15±0.46d 2.51±0.49e 5.705* 153.3***
Harvest Index % 65.2±40.0abcf 86.9±67.8bdg 49.7±34.3ceh 73.4±51.1ade 87.8±61.4agh 47.0±39.2cf 0.292 n.s 7.713**
109
attributes in better measure than the 5 and 10 t ha-1
treatments, whereas seed diameter and seed
thickness were maximum in 10 t ha-1 treatments.
Even though, VC treated plants given significantly
higher fruit yield, the harvest index (HI) values of
VC treatments are not significantly higher than that
of the IF treated plants. This is because HI is a ratio
of mass of fruits and mass of the above-ground
biomass. In VC treated plants both are higher than
in IF treated plants leading to similar HI. However,
different dose of nutrient application showed
significant influence on HI. Among all the
treatment, the maximum HI was recorded with 7.5 t
ha-1
VC equivalent IF treatment followed by 7.5 t
ha-1
VC treatment; more than 12% higher HI was in
the 7.5 t ha-1
IF treatment.
4. Conclusions
The impact of vermicompost (VC) generated
from the pernicious, allelopathic weed Ipomoea
carnea on germination, growth and fruition of a
botanical species was studied. Response of plants to
the vermicompost was compared with that of an
inorganic fertilizer (IF) which had all the main
macro and micro – nutrients in concentrations
equivalent to the ones present in the VC, and a
control without any nutrient supplement. In this
experiment, nutrient has been supplemented at three
doses, 5, 7.5 or 10 tons per hectare (t ha-1
) and a
legume species, cluster bean (Cyamopsis
tetragonoloba) was used.
In this study, the plant growth in terms of
stem diameter, shoot length, shoot mass, number of
leaves, and leaf pigments were significantly
improved with VC application than with IF. In
addition, VC application enhanced root nodule
formation, and permitted lesser number of stunted
plants; whereas, the IF application suppressed the
root nodulation in the experimental plants. The
plant growth was significantly improved with
increasing dose of both VC and IF application.
However, the yield was maximum in 7.5 t ha-1
VC/IF treatments than the 5 and 10 t ha-1
treatments. Moreover, the root elongation was also
suppressed by excess nutrient application in both
VC and IF treatments. Overall results show that
vermicompost generated from ipomoea had positive
impact on the growth and yield of the plant studied.
In terms of germination, vermicast showed
suppression effect, indicating the presence of some
of the germination inhibitory components in
vermicast.
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cyamopsidis. Ann. Plant Prot. Sci. 3, 171-172.
112
Supplementary material
Table 4A. Stem diameter (mm) of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
1.0 1.6 2.1 4.3 5.2 7.1 8.3 12.7 13.7 16.2 16.9 16.9
7.5 t ha-1
1.3 2.1 2.9 3.9 7.6 9.0 11.5 15.3 15.5 22.3 22.5 22.6
10 t ha-1
1.4 2.3 3.2 5.6 8.1 10.1 12.2 17.8 19.2 23.7 24.0 24.1
Inorganic
fertilizer
5 t ha-1
1.2 2.2 2.4 4.5 5.0 6.7 8.6 11.5 13.2 16.6 16.8 16.8
7.5 t ha-1
1.5 2.3 2.7 4.8 5.9 7.9 9.3 14.4 17.4 18.1 18.3 18.4
10 t ha-1
1.5 2.7 2.9 5.7 6.3 8.5 10.2 16.4 20.3 22.7 22.9 22.9
Control Nil 0.7 1.8 2.0 2.3 4.5 5.2 5.8 6.1 1.2 10.8 11.1 11.2
Table 4B. Shoot length (cm) of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
4.8 9.5 14.6 23.5 25.2 28.5 37.6 53.1 88.0 130.5 1394.1 144.5
7.5 t ha-1
7.5 11.7 19.7 24.8 32.6 40.5 51.4 59.7 90.2 190.8 192.8 197.6
10 t ha-1
11.5 12.5 21.2 27.4 37.8 44.2 59.8 72.9 115.6 220.1 225.4 231.6
Inorganic
fertilizer
5 t ha-1
6.5 10.5 15.8 17.5 23.7 28.0 34.8 54.0 80.2 122.5 132.8 138.6
7.5 t ha-1
7.5 11.7 16.3 19.1 25.9 29.5 36.9 48.9 94.3 140.5 153.9 162.7
10 t ha-1
10.5 13.5 18.8 21.4 27.2 29.0 41.3 47.2 103.6 175.8 180.2 191.4
Control Nil 3.5 11.4 11.8 12.5 18.0 21.8 23.0 26.0 34.5 42.0 59.3 76.8
Table 4C. Root length (cm) of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
4.2 16.1 18.5 25.5 27.6 29.2 31.9 32.8 34.6 51.4 54.8 55.0
7.5 t ha-1
5.5 15.4 19.6 21.5 23.6 24.5 25.7 27.1 29.6 37.5 38.5 41.4
10 t ha-1
6.8 21.5 22.5 24.3 25.7 26.2 28.1 29.6 31.6 34.2 36.8 38.3
Inorganic
fertilizer
5 t ha-1
4.9 13.1 19.5 27.3 29.7 33.0 35.9 38.7 41.8 43.1 45.4 47.7
7.5 t ha-1
5.5 13.4 21.4 28.2 30.2 33.2 37.6 40.7 41.5 42.8 45.3 47.2
10 t ha-1
7.6 15.7 18.2 22.3 24.2 25.8 27.1 28.3 32.3 36.8 38.1 39.6
Control Nil 3.8 13.7 14.0 23.1 29.1 29.2 37.4 42.0 45.7 47.5 49.3 50.2
113
Table 4D. Number of leaves of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
03 03 04 14 19 33 37 48 55 87 92 98
7.5 t ha-1
03 04 07 19 24 34 42 60 72 105 117 120
10 t ha-1
03 04 07 20 28 43 47 65 81 118 125 134
Inorganic
fertilizer
5 t ha-1
03 03 04 17 21 33 39 58 62 72 85 92
7.5 t ha-1
03 04 07 18 24 33 41 52 68 82 98 108
10 t ha-1
03 04 08 21 27 28 48 59 82 99 107 112
Control Nil 03 03 04 09 15 17 21 25 28 31 35 42
Table 4E. Number of nodules of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
Nil 8 11 12 15 20 26 42 38 42 58 61
7.5 t ha-1
Nil 12 13 10 21 24 29 24 42 57 72 90
10 t ha-1
Nil 12 15 15 26 33 35 52 60 59 86 129
Inorganic
fertilizer
5 t ha-1
Nil 3 5 8 9 25 28 39 42 28 29 32
7.5 t ha-1
Nil 12 15 22 27 34 31 30 43 49 42 50
10 t ha-1
6 8 7 4 14 29 18 35 29 31 30 37
Control Nil Nil 0 7 8 12 14 19 11 11 9 16 14
Table 4F. Shoot dry weight (g) of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.031 0.102 0.459 0.981 2.147 4.446 13.067 21.07 25.93 47.58 49.16 51.46
7.5 t ha-1
0.036 0.181 0.367 0.758 2.668 5.755 15.45 24.81 29.18 83.53 88.04 89.31
10 t ha-1
0.054 0.134 0.648 1.065 4.093 7.033 19.31 27.58 37.13 92.62 99.27 102.56
Inorganic
fertilizer
5 t ha-1
0.082 0.120 0.493 0.820 2.105 4.223 12.30 19.88 24.79 40.21 41.72 42.92
7.5 t ha-1
0.047 0.136 0.355 0.647 2.485 5.261 14.94 20.38 27.34 56.24 57.93 59.64
10 t ha-1
0.008 0.094 0.539 0.602 3.827 6.875 17.11 24.11 34.60 74.55 79.62 82.81
Control Nil 0.032 0.109 0.205 0.395 0.955 1.490 4.572 6.799 12.11 41.50 44.08 45.32
114
Table 4G. Root dry weight (g) of cluster bean at different weeks
Table 4H. Leaf chlorophyll a (mg g-1
) content of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.398 0.779 1.095 1.503 1.716 1.791 1.645 1.798 1.823 1.925 2.092 2.167
7.5 t ha-1
0.414 1.642 1.955 2.115 2.326 2.401 2.578 2.774 2.823 2.890 3.172 3.211
10 t ha-1
0.543 1.795 2.093 2.375 2.583 2.741 2.874 3.108 3.195 3.284 3.376 3.452
Inorganic
fertilizer
5 t ha-1
0.378 0.914 1.218 1.649 1.789 1.589 1.674 1.699 1.718 1.825 1.959 1.990
7.5 t ha-1
0.307 1.458 1.541 1.632 1.735 1.77 1.823 2.236 2.419 2.497 2.556 2.691
10 t ha-1
0.541 1.618 1.481 1.781 1.974 2.122 2.249 2.364 2.592 2.614 2.628 2.717
Control Nil 0.103 0.465 0.64 0.907 1.128 1.164 1.177 1.193 1.201 1.224 1.238 1.316
Table 4I. Leaf chlorophyll b (mg g-1
) content of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.102 0.627 0.948 1.06 1.18 1.254 1.635 1.806 1.902 2.132 2.160 2.207
7.5 t ha-1
0.148 0.757 0.978 1.209 1.497 1.712 1.964 2.024 2.100 2.195 2.222 2.296
10 t ha-1
0.291 0.999 1.167 1.249 1.591 1.854 2.072 2.168 2.131 2.205 2.234 2.474
Inorganic
fertilizer
5 t ha-1
0.179 0.304 0.963 1.118 1.244 1.418 1.64 1.823 1.825 2.014 2.060 2.097
7.5 t ha-1
0.141 0.475 1.021 1.393 1.571 1.846 1.91 2.116 2.176 2.182 2.220 2.278
10 t ha-1
0.374 0.52 1.134 1.586 1.618 1.987 2.128 2.351 2.404 2.481 2.517 2.568
Control Nil 0.032 0.155 0.223 0.504 1.612 1.935 1.065 1.409 1.510 1.532 1.544 1.451
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.005 0.024 0.039 0.066 0.125 0.607 0.984 1.642 3.094 4.751 4.882 5.024
7.5 t ha-1
0.001 0.026 0.042 0.086 0.276 0.847 1.446 2.044 3.916 5.983 6.034 6.248
10 t ha-1
0.005 0.019 0.057 0.084 0.283 0.621 1.349 2.323 6.800 9.019 9.275 11.937
Inorganic
fertilizer
5 t ha-1
0.000 0.022 0.040 0.072 0.174 0.501 0.728 1.847 2.171 4.192 4.301 4.605
7.5 t ha-1
0.003 0.024 0.042 0.089 0.226 0.738 1.203 2.403 6.064 7.652 7.824 8.102
10 t ha-1
0.000 0.012 0.029 0.064 0.198 0.515 1.004 1.077 5.903 6.061 6.489 6.841
Control Nil 0.001 0.014 0.016 0.029 0.102 0.256 0.289 0.981 1.763 2.929 3.160 3.214
115
Table 4J. Leaf total chlorophyll (mg g-1
) content of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.500 1.406 2.043 2.563 2.896 3.045 3.280 3.604 3.725 4.057 4.252 4.374
7.5 t ha-1
0.562 2.399 2.933 3.324 3.823 4.113 4.542 4.798 4.923 5.085 5.394 5.507
10 t ha-1
0.834 2.794 3.26 3.624 4.174 4.595 4.946 5.276 5.326 5.489 5.610 5.926
Inorganic
fertilizer
5 t ha-1
0.557 1.218 2.181 2.767 3.033 3.007 3.314 3.522 3.543 3.839 4.019 4.087
7.5 t ha-1
0.448 1.933 2.562 3.025 3.306 3.616 3.733 4.352 4.595 4.679 4.776 4.969
10 t ha-1
0.915 2.138 2.615 3.367 3.592 4.109 4.377 4.715 4.996 5.095 5.145 5.285
Control Nil 0.135 0.62 0.863 1.411 2.740 3.099 2.242 2.602 2.711 2.756 2.782 2.767
Table 4K. Leaf carotenoids (mg g-1
) content of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.101 0.139 0.197 0.261 0.292 0.314 0.335 0.364 0.371 0.398 0.405 0.424
7.5 t ha-1
0.126 0.235 0.289 0.322 0.357 0.398 0.423 0.456 0.477 0.483 0.491 0.512
10 t ha-1
0.166 0.244 0.302 0.384 0.416 0.427 0.438 0.479 0.489 0.503 0.528 0.547
Inorganic
fertilizer
5 t ha-1
0.16 0.192 0.231 0.249 0.276 0.288 0.291 0.301 0.312 0.333 0.348 0.361
7.5 t ha-1
0.104 0.151 0.247 0.275 0.291 0.317 0.341 0.366 0.371 0.382 0.400 0.412
10 t ha-1
0.107 0.266 0.382 0.416 0.427 0.435 0.442 0.456 0.482 0.499 0.529 0.530
Control Nil 0.102 0.122 0.136 0.158 0.164 0.191 0.203 0.218 0.224 0.236 0.248 0.261
116
COMPARATIVE EFFICACY OF VERMICOMPOSTED
PAPER WASTE AND INORGANIC FERTILIZER ON
SEED GERMINATION, PLANT GROWTH AND
FRUITION OF CLUSTER BEAN (CYAMOPSIS
TETRAGONOLOBA)
Chapter
8
117
A paper based on this chapter has been published in
Journal of Applied Horticulture, 16 (1), 40 – 45, 2014.
CChhaapptteerr 88
Comparative efficacy of vermicomposted paper waste and
inorganic fertilizer on seed germination, plant growth and
fruition of cluster bean (Cyamopsis tetragonoloba)
Abstract
The aim of the present study was to assess the influence of vermicompost generated from the paper waste
spiked with cow dung slurry on the germination, plant growth and fruition of cluster bean (Cyamopsis
tetragonoloba). Two kinds of treatments were studied: (i) vermicast was applied to the soil at the rates of 5,
7.5, 10 t ha-1
, (ii) amounts of essential nutrients equivalent to those present in the vermicast treatments in
inorganic form was amended to the soil. There was a control with only soil without any nutrient supplement.
The finding is in contrast to the reports on the beneficial impacts of vermicast on plant growth. In the present
study the inorganic fertilizer treatment exhibited better seed germination and plant growth than the
equivalent vermicast treatments. The results indicate that the dose of vermicompost used in the present study
was not sufficient to satisfy the nutrient demand of plant species studied. Additional fertilization would have
improved the crop productivity.
1. Introduction
Paper waste generation has been continually
increasing over the past years due to increasing
population, industrialization, urbanization and
literacy. In India, the paper consumption is about
8.5 kg per capita per year and it is 0.81-5.8 % of
municipal solid waste (MSW) (Gupta and Garg,
2009; www.indiastat.com). Due to the absence of
waste segregating practices, the paper waste is
dumped along with all other kinds of waste in open
and poorly managed landfills, which is very
common practice in most of the cities in India. The
improper disposal of this degradable waste may
lead to long term threat to the environment and
public health, such as the risk of ground water
pollution due to leachate seepage, fugitive
greenhouse gas emission contributing to climate
change and odor pollution caused by non-methane
organic compounds, which is direct harassment to
adjacent communities (Zhang et al., 2012). Also,
open dumping of wastes facilitates the breeding for
disease vectors such as flies, mosquitoes,
cockroaches, rats, and other pests (CPCB, 2000).
Paper and cardboard have a relatively high
heating value, similar to wood, and this energy
utilized via incineration can be transformed into
electricity (Villanueva and Wenzel, 2007).
However, incineration is not very much practiced in
India due to lack of awareness and absence of waste
segregating practices. The paper waste when mixed
with other moist organic waste and inert material
reduces its calorific value (Negi and Suthar, 2013).
In recent years, due to shortage of raw material,
waste paper is preferred for paper production. Also,
recycling of paper consumes only 40% of the
energy in comparison to the process based on other
raw materials (Gupta et al., 1998). However, the
paper recycling industries prefer to use imported
waste paper because of its better quality in terms of
fibre strength and also due to inadequate domestic
118
supply owing to the unorganized collection of
waste paper within the country. In addition, the
yield from imported waste paper can be as high as
90%, whereas the available waste paper in India
gives yield less than 50% due to shorter fibre length
(IPMA, 1996). Nevertheless, if waste papers are
segregated at the source itself, it could be the input
material for paper recycling units. However, due to
lack of efficient waste management service, the
paper waste invariably finds its way to the MSW at
the end.
The huge generation of these paper wastes
can be treated by vermicomposting which convert
the waste into useful end product that can be used
as a soil amendment (Sinha et al., 2010). Unlike
recycling and incineration, the biological
composting process not affected either by quality of
waste paper or when mixed with other organic
wastes. Processing of paper waste through
vermicomposting may provide an answer to the
minimization of waste accumulation and also to
widespread deteriorated agricultural land due to
rampant use of inorganic fertilizers. It is well
established that vermicompost application have
beneficial impact on soil physical, chemical and
biological properties and can increase the
germination, plant growth and yield in both natural
and agricultural ecosystem (Edwards and Bohlen,
1996). These beneficial effects have been attributed
to improvement in soil properties and structure, to
greater availability of mineral nutrients to plants
(Edwards, 1998). In addition to this, vermicompost
contain plant growth regulating components,
including plant growth hormones and humic acids
that are reported to be responsible for promoting
germination, growth and yields of plants, in
response to vermicompost applications or
substitutions, independent of the nutrients they
contain (Tomati et al., 1988; Muscolo et al., 1999;
Atiyeh et al., 2002; Arancon et al., 2003, 2006).
However, the vermicompost generated from
waste paper causes apprehension towards the
beneficial impact on plant growth due to the low
nutrient content of this substrate. Therefore, attempt
has been taken to investigate the beneficial impact
of vermicompost generated from the paper waste
(VC) on the germination, plant growth and yield of
cluster bean, a vegetable crop. This plant has
chosen due to their drought tolerance which reduces
the error due to the other environmental factors.
Moreover as it is a leguminous plant, influence of
vermicast and inorganic fertilizers on nodules
formation and its growth can be revealed. In
addition, to evaluate the possible non-nutrient (i.e.
hormones and other growth regulating components)
dependent effect of vermicast over inorganic
fertilizers, all essential nutrients present in the
vermicast were supplied in inorganic form (IF) to
the plants, and the response of plants to the
different fertilizers is briefed in this paper.
2. Materials and methods
2.1. Study area
This experiment was conducted at
Pondicherry University, Puducherry, India,
located on the east coast of Indian peninsula
(11°56’N, 79°53’E). The climate of the
experimental site is typical maritime of tropical
climate with a disymmetric rainfall. The
average annual rain fall is about 1300 mm with
57.25 mean rainy days, and around 60% of the
total rainfall is received during period of
October to December through the north–east
monsoon.
2.2. Treatments
The experiment was set up in 49 liter volume
wooden containers (40 cm height with surface area
35 x 35 cm), lined with high-density polyethylene
(HDPE) sheets. These containers were filled with
low fertile barren land soil collected inside the
Pondicherry University campus to reduce the errors
due to previous soil practices. The experimental
soil was characterized as sandy loam soil and its
physico-chemical properties are shown in Table 1.
119
Table 1. Chemical and physical properties of
vermicast and soil used in the study.
BDL – Below detection limit
The experiment was conducted during the Kharif
season which is best time for sowing the cluster
beans in south India. The Pusanavabahar variety
was used which is locally available in the
experimental area. The vermicompost was
generated from the paper waste spiked with cow
dung slurry by employing an epigeic species,
Eudrilus eugeniae Kinberg. The VC was applied to
the plant growth containers at the rate of 5, 7.5 and
10 t ha-1
. In another set, an equivalent amount of all
major and minor nutrients present in vermicompost
was supplied as inorganic chemical form to check
the efficiency of vermicast over the inorganic
fertilizer.
In the IF treatment, the primary nutrients N, P
and K, secondary nutrients Ca, Mg and S, and
micronutrients of Fe, Mn, Cu, Zn, B, Mo and Cl
were applied to an equivalent amount of 5, 7.5 and
10 t ha-1
VC treatment. The chemical fertilizers
were applied in the form of urea, di-ammonium
phosphate, potash, CaCO3, MgO, Na2B4O7, CuSO4,
FeSO4, MnSO4 and ZnCl2. Besides these
treatments, one more set was maintained without
any supplementation, as control i.e only soil. The
nutrients were supplied in two phases. The first
phase was at the time of sowing which comprised
half of the total nutrients. The second
supplementation of nutrients was done at the time
of flowering of plants.
2.3. Germination, plant growth and yield
characteristics
Two seeds per container, totally 72 seeds per
treatment were sown in all the containers. Seeds
were considered germinated when they exhibited
radical extension of >3 mm. Counts of germinated
seeds were made daily up to eight days to
determine the germination rate in terms of
germination percentage (GP) and germination value
(GV) by method described by Djavanshir and
Pourbeik (1976). The GV is used as a comparative
index to statistically assess the effects of the
treatments. After germination, one plant for each
compartment was maintained by removing excess
plant grown from the sown seeds. A separate
nursery was also maintained with all VC and IF
treatments. Healthy plants from the nursery were
transplanted to containers where seeds failed to
germinate. Adequate amount of water was provided
during the experiment. Deweeding was done
manually. In few instances when pests were seen,
organic pesticides such as neem extract and cow
urine were used.
The plant height, length of shoot and root,
Variables Concentration
Vermicast Soil
Chemical properties
pH 7.83±0.05 6.30±0.10
Total organic carbon g kg-1
259.6±8.2 8.87±0.02
Total Nitrogen g kg-1
11.7±0.4 2.66±0.02
Plant available form of
Phosphorus mg kg -1
71.40±3.31 0.41±0.01
Potassium g kg-1
1.98±0.10 0.40±0.00
Sulphur mg 100g-1
0.40±0.04 0.54±0.01
Calcium g kg-1
15.9±1.4 8.27±0.01
Magnesium g kg-1
7.13±0.07 0.09±0.02
Boron mg kg-1
7.28±0.09 26.9±1.2
Copper mg kg-1
16.9±1.0 5.08±0.15
Iron mg kg-1
208.3±11.4 59.9±2.9
Manganese mg kg-1
63.8±1.8 45.1±2.2
Zinc mg kg-1
94.1±2.2 55.0±2.6
Molybdenum mg kg-1
BDL BDL
Physical properties
Dry weight % 49.6±1.9 94.7±0.1
Bulk density g cm-3
0.24±0.01 1.28±0.00
Particle density g cm-3
1.21±0.03 2.70±0.12
Water-holding capacity % 118.0±11.2 36.9±2.4
Electrical conductivity
mmhos cm-1
2.96±0.04 0.12±0.02
Total porosity % 80.08±1.0 53.1±2.0
Air filled porosity % 68.14±1.2 46.5±2.1
120
Table 2. Amount of inorganic fertilizer applied equivalent to vermicast treatment.
Nutrients Form in which
applied
Mass %
of nutrient
in applied
compound
10t ha-1
7.5t ha-1
5t ha-1
Concentra
tion of
nutrient in
VC
(kg ha-1
)
Amount
of IF
applied
(kg ha-1
)
Concentration
of nutrient in
VC
(kg ha-1
)
Amount
of IF
applied
(kg ha-1
)
Concentration
of nutrient in
VC
(kg ha-1
)
Amount
of IF
applied
(kg ha-1
)
Nitrogen CH4N2O 46.65 30.39 65.15a 22.79 48.86
a 15.19 32.57
a
Phosphorus (NH4)2HPO4 23.45 35.40 150.9 26.55 113.2 17.70 75.45
Potassium KCl 52.44 9.806 18.70 7.354 14.02 4.903 9.348
Sulphur 26.93 0.020 Nilb 0.015 Nil
b 0.010 Nil
b
Calcium CaCO3 40.04 78.98 197.2 59.23 147.9 39.49 98.62
Magnesium MgO 60.3 35.36 58.63 26.52 43.97 17.68 29.31
Boron Na2B4O7.10H2O 11.34 0.036 0.318 0.027 0.239 0.018 0.159
Copper CuSO4.5H2O 25.45 0.084 0.329 0.063 0.247 0.042 0.165
Iron FeSO4.7H2O 20.09 1.033 5.141 0.774 3.855 0.516 2.570
Manganese MnSO4. H2O 32.5 0.316 0.973 0.237 0.730 0.158 0.487
Zinc ZnCl2 47.97 0.466 0.972 0.350 0.729 0.233 0.486 a The total amount of N applied in the form of urea and di-ammonium phosphate is equal to the concentration of N applied in the
vermicast treatment. b The total amount of sulphate applied with other elements equalizes the sulphate in applied vermicast.
number of leaves, stem diameter, number of
nodules present in the root and their size, biomass
of shoot and root and its dry weight were recorded
with randomly collected samples at each week.
Leaf chlorophyll a, b and carotenoids content were
estimated photometrically by using N,N-di-Methyl
formamide (DMF) as extractent (Moran and Porath,
1980; Wellburn, 1994). Throughout the study, the
disease incidence, and number of flowers produced
were recorded.
2.3. Analytical methods
The analytical methods were the same as
detailed in section 2.3 of chapter 6.
2.4. Statistical analysis
One-way ANOVA and post hoc LSD tests
were employed for determining significance of
difference between the results. The statistical
analysis was carried out using SPSS software
(version 16). The level of significance was at p <
0.05.
3. Results and discussion
3.1. Seed germination
In both VC and IF treatments, maximum GV
was observed on fourth day since the experiment
began and then a decreasing trend was observed as
the days progressed. The results of GV did not
show any significant difference with different
forms nor amount of nutrients applied. However,
both forms of fertilizer and amount of nutrient
applied showed significantly higher GV than the
control (p<0.001), and it was more than 10 folds
higher in most of the cases (Table 4). In the VC
treatment maximum GP of 88.6% was observed
with both 7.5 and 10 t ha-1
treatment and maximum
GV of 40 was also recorded in these treatments.
The VC treatment at dose of 5 t ha-1
showed lowest
GP of 74.3% among both VC and IF treatments. In
the IF treatment, an increasing trend in GP was
observed with increasing dosage of fertilizer
application. In this treatment, GP of 88.6, 91.4 and
94.3 % recorded with 5, 7.5 and 10 t ha-1
treatment
on the eighth day and, a maximum GV of 42.9 was
in 7.5 t ha-1
treatments. With all the dose of nutrient
121
application, the VC had shown lesser germination
rate than their respective inorganic fertilizer
treatments (Table 3).
Other studies on influence of VC on seed
germination either showed inhibitory or stimulation
effect with low concentration of VC substitution in
growth media. The discrepancy in the response to
vermicompost depends on plant species which
reacts differently to the concentration of
vermicompost application (Atiyeh et al., 2000;
Edwards et al., 2004). The different organic
substrate used for vermicomposting also changes
the quality of vermicast (Zaller, 2007). Ievinsh
(2011) reported inhibitory effect similar to the
present study on beetroot, beans and pea at low
dose of manure-based vermicompost (5-10%), but
germination dramatically increased with the
increase of vermicompost concentration. The
higher germination in IF treatments may be due to
nitrate which would have been converted from the
urea applied as nitrogen source. These constituents
are breakers of dormancy and also stimulator of
germination (Egley and Duke, 1985; Hihorst and
Karssen, 2000). Since the nitrate readily leaches
from surface soil by irrigation water, the
concentration of nitrate would have declined as the
days progressed, and this may be the reason for
higher germination at initial days followed by steep
inclination till the end.
3.2. Plant growth
There was a differential impact of the vermicast
and inorganic fertilizer treatment on all the
morphological parameters studied (Table 3). The
plants which were treated with IF grew faster than
the VC treated plants. The IF treatment equivalent
to 10 t ha-1
VC recorded higher stem diameter,
length, fresh and dry weight of shoot, number of
leaves, number and size of nodules than VC
treatment and control. The highest of these
parameters: 14.4 mm stem diameter, shoot length of
134 cm, 72 numbers of leaves, 182.1 g of shoot
fresh weight and 52.4 g shoot dry weight was
recorded in the plants treated with IF treatment
equivalent to 10 t ha-1
paper waste VC. Plants
treated with VC at the rate of 10 t ha-1
showed a
maximum of 14.1 mm of stem diameter; 121 cm
shoot length, 65 numbers of leaves, 180 g of shoot
fresh weight and 57.3 g of shoot biomass.
Many authors have reported that high dose of
inorganic fertilizer treatments induce depression in
nodules formation in soybean, chick pea and lupins
(van Schreven, 1959; Carroll and Gresshoff, 1983;
Table 3. Germination value (GV) and germination percentage (GP) of the seeds of cluster bean as influenced
by paper waste vermicast and inorganic fertilizers. Results which do not differ significantly (LSD test; p
<0.05) carry at least one character in the superscript which is common.
Treatment Amount Fourth day Fifth day Sixth day Seventh day Eighth day
GV GP GV GP GV GP GV GP GV GP
Vermicompost
5t ha-1 a
24.70 62.86 17.27 65.71 15.33 74.29 11.26 74.29 8.62 74.29
7.5 t ha-1 b
40.00 80.00 27.46 82.86 20.41 85.71 16.01 88.57 12.26 88.57
10 t ha-1b
40.00 80.00 29.38 85.71 21.79 88.57 16.01 88.57 12.26 88.57
Inorganic
fertilizer
equivalent to
the VC
5t ha-1 b
40.00 80.00 27.46 82.86 20.41 85.71 16.01 88.57 12.26 88.57
7.5 t ha-1 b
42.91 82.86 29.38 85.71 20.41 85.71 14.99 85.71 13.06 91.43
10 t ha-1 b
40.00 80.00 33.44 91.43 24.70 94.29 18.14 94.29 13.89 94.29
Control Nil c 2.50 20.00 2.64 25.71 2.74 31.43 4.26 45.71 3.47 47.14
122
Table 4. Plant growth, leaf pigments, flowering, disease incidence, and retarded growth/death in cluster bean plants as impacted by paper waste
vermicast or inorganic fertilizers. Results which do not differ significantly (LSD test; p <0.05) carry at least one character in the superscript which is
common.
*p<0.05,
**p<0.01,
***p<0.001,
n.s - not significant.
Parameters
Vermicompost at dose Inorganic fertilizers at dose
Control
F-value
5 t ha-1
7.5 t ha-1
10 t ha-1
5 t ha-1
7.5 t ha-1
10 t ha-1
Type of
fertilizer Amount
Plant growth
Stem diameter (mm) 10.9±0.8a 12.8±0.9
bc 14.1±1.2
de 11.9±0.9
ab 13.1±1.4
cd 14.4±2.0
e 11.2±1.7
a 5.634
** 22.55
***
Shoot length (cm) 96.8±4.1a 118.0±10.6
b 121.0±7.9
b 108.4±7.3
c 119.1±4.7
b 134.3±10.9
d 76.8±5.9
e 54.58
*** 97.36
***
Root length (cm) 59.2±9.2a 72.1±14.9
b 83.4±6.1
c 55.7±11.0
ad 89.1±15.5
c 50.1±7.8
d 50.2±8.4
d 7.178
** 14.88
***
Number of leaves 58.8±4.2a 62.2±3.2
ab 64.8±5.7
bc 51.2±6.0
d 68.3±5.4
ce 71.8±4.0
e 42.0±5.3
f 36.49
*** 66.51
***
Number of nodules 27.8±10.2a 40.9±13.9
b 45.3±8.4
b 38.4±11.8
b 38.3±6.6
b 37.1±14.0
b 13.5±6.0
c 23.45
*** 19.26
***
Shoot dry weight (g) 37.1±3.8ab
47.6±5.7c 57.3±6.5
d 39.8±4.0
b 43.2±3.8
be 52.4±6.2
f 45.3±5.2
ce 0.699
n.s 36.92
***
Root dry weight(g) 4.78±0.29a 6.82±0.20
b 7.22±0.55
bc 7.53±0.62
c 9.39±0.86
d 12.7±1.7
e 3.21±0.58
f 74.94
*** 35.04
***
Leaf pigments
Chlorophyll a (mg g-1
) 1.83±0.12a 1.92±0.15
a 1.87±0.10
a 1.84±0.08
a 2.24±0.14
b 2.29±0.12
b 1.32±0.05
c 98.03
*** 57.51
***
Chlorophyll b (mg g-1
) 1.61±0.10ab
1.63±0.11bc
2.05±0.17b 1.51±0.15
ad 2.12±0.19
c 2.28±0.11
e 1.45±0.04
d 14.77
*** 52.62
***
Total chlorophyll (mg g-1
) 3.44±0.12ab
3.55±0.22a 3.92±0.22
c 3.35±0.17
b 4.36±0.23
d 4.57±0.17
e 2.77±0.04
f 45.88
*** 63.60
***
Carotenoids (mg g-1
) 0.29±0.04a 0.46±0.06
b 0.46±0.08
b 0.36±0.10
c 0.36±0.07
c 0.37±0.05
c 0.26±0.02
a 13.17
*** 15.32
***
Flowering
Number of flowers 1.42±1.05a 3.08±1.79
b 3.61±1.99
bc 3.17±1.70
b 4.08±2.47
c 4.39±2.56
c 0.72±1.02
a 34.90
*** 27.57
***
Disease incident, plant death and stunted plants
Number of infected plant 19a 28
a 21
a 8
a 14
a 12
a 16
a 2.221
n.s 0.378
n.s
Number of plant died Nila 1
a Nil
a 1
a Nil
a Nil
a Nil
a 0.162
n.s 0.490
n.s
Number of stunted plant Nila 2
a 1
a 1
a 6
a 11
a 11
a 1.276
n.s 0.749
n.s
123
Harper and Gibson, 1984; Nie, 1989). The finding
of the present study contradicts with the previous
reports on suppression of nodules formation with IF
treatment. There was no significant difference in
the number and size of nodules between VC and IF
treatments. The results indicates that there was no
suppression or stimulation of nodules formation
with both VC and IF treatment. The reason may
that very low amount of nutrient has been supplied
to the plants in both VC and IF treatments. In
addition to this, slow releasing property of
vermicast might have reduced the nutrient
availability to the plants. In the VC treated
containers, it was observed that the applied VC did
not disintegrate till the end of the experiment
period. This stable nature of VC might have slow
down the nutrient release to plants. It might be the
reason for lower plant growth in the VC treatment
than the equivalent IF treatment.
3.3. Photosynthetic pigments
The photosynthetic pigments such as
chlorophyll and carotenoid in the leaves of plants
amended with vermicast and inorganic fertilizer
highly correlated with amount of nutrient applied
(Table 3). In general, IF application was more
effective than respective VC treatment. Lower rate
of nutrient application and their poor availability
due to the slower releasing property of vermicast
may be the reason for lower photosynthetic
pigments in this treatment compared to IF treated
plants. In the VC treatment, the maximum total
chlorophyll content of 3.92 mg g-1
was recorded
with the dose of 10 t ha-1
followed by 3.55 and 3.44
mg g-1
with 7.5 and 5 t ha-1
treatments respectively.
In the case of IF treatment, the maximum total
chlorophyll content of 4.57 mg g-1
was recorded in
10 t ha-1
treatment followed by 4.35 and 3.35 mg g-1
with 7.5 and 5 t ha-1
IF treatment. The control
treatment showed lowest chlorophyll content of
2.77 mg g-1
at the end of the experiment.
Carotenoids pigment exhibited similar trend of
results. The increasing nutrient availability with
increasing dose of fertilizer application can be
attributed to the formation of leaf pigments
(Mengel and Kirkby, 1987; Shadchina and
Dmitrieva, 1995; Ruza, 1996; Tejada et al., 2007).
3.4. Disease incidence, plant death and stunted
growth
In the first two months of the experimental
period many plants were infected with bacterial
blight (Xanthomonas campestris), and alternaria
blight (Alternaria spp) and infested with whiteflies
(Bemisia tabaci). As a consequence some plants
died and some did not exhibit normal growth
(Table 3). The observed disease incidence and plant
mortality was not significantly differing between
different forms of nutrient application (vermicast
and inorganic fertilizer). The number of stunted
plants was maximum in the beginning of the
experiment. After a month, all the plants grew well.
Inorganic treatment showed more stunted plants in
comparison to the VC treatment. In IF treatment
totally 18 stunted plants were recorded, whereas in
the case of VC treatment at different doses, two or
less stunted plants were recorded.
3.5. Effect on flowering
The IF treated plants were produced more
number of flowers in comparison to the equivalent
VC treated plants (p<0.001; Table 5). A maximum
was recorded with 10 t ha-1
treatment with both VC
and IF treatments. Increasing dose of both fertillizer
application showed the trend with maximum
number of flowers: 10 t ha-1
> 7.5 t ha-1
> 5 t ha-1
>
control. The plants grown in the soil treated with
10 t ha-1
of both VC and IF showed early flowering
than the other treatments. Increased in availability
of nutrient in this treatment could be the reason for
the higher and early flowering in this treatment.
3.6. Effect on yield
In the VC, IF treated plants and control no
vegetables were produced. The reason may be due
to inadequate availability of minor nutrients to the
plants in all the treatments. In this study, low fertile
124
soil collected from barren land was used to
minimize the errors due to the previous soil
practices in the experimental results. The soil was
characterized as very low nutrient soil. Therefore,
the growth and yield of plants relied on the applied
nutrient either in the form of VC or IF. The minor
nutrient applied would have been exhausted thereby
impeding the fruiting in all the treatments.
4. Conclusions
The results obtained from this experiment
shows that application of VC generated from the
paper waste had no beneficial impact on growth of
cluster bean plant. Moreover, the IF treatment
exhibited better seed germination and plant growth
than the equivalent VC treatment. This indicates
that the slow releasing property of VC might have
led to depletion of nutrient availability to the plants.
In this experiment, increasing dose of both
vermicast and inorganic fertilizer improved the
germination rate and plant growth parameters.
However, the application of inorganic fertilizer did
not suppress the root nodules, which is
contradictory to the previous reports (van Schreven,
1959; Nie, 1989). There was no significant
difference in the number and size of nodules with
different treatments. In addition to this, the plants
did not fructify in all the treatments. The low fertile
experimental soil and insufficient nutrient
application might have impeded the production of
vegetables in the present study.
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Supplementary material
Table 4A. Stem diameter (mm) of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
1.2 1.6 1.9 2.5 3.9 5.1 5.6 6.3 9.7 10.2 10.9 10.9
7.5 t ha-1
1.6 1.7 1.8 3.8 4.1 5.6 5.8 6.7 11.2 11.9 12.3 12.8
10 t ha-1
1.7 1.8 2.2 3.3 3.8 3.9 5.3 8.6 11.9 12.6 13.3 14.1
Inorganic
fertilizer
5 t ha-1
1.7 1.8 2.1 2.9 3.2 4.4 5.3 6.1 10.1 11.1 11.4 11.9
7.5 t ha-1
1.6 1.6 1.8 3.1 4 4.1 5.8 6.1 10.4 11.5 11.9 13.1
10 t ha-1
1.2 1.5 1.2 3.7 5.5 8.1 8.6 9.1 11.4 13.1 13.6 14.4
Control Nil 0.7 1.8 2.0 2.3 4.5 5.2 5.8 6.1 1.2 10.8 11.1 11.2
Table 4B. Shoot length (cm) of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
14.2 22.3 23.1 25.4 26.5 33.7 41.3 55.7 63.5 73.6 89.1 96.8
7.5 t ha-1
14.1 21.5 21.5 23.4 26.1 39.0 62.4 65.2 77.5 82.3 103.6 118
10 t ha-1
13.1 21.0 14.6 18.6 27.5 33.0 52.5 72.1 98.7 105 113 121
Inorganic
fertilizer
5 t ha-1
11.5 17.1 18.5 22.0 24.6 33.4 34.9 38.0 56.0 67.0 95 108
7.5 t ha-1
14.4 18.5 20.8 25.9 32.0 38.0 48.3 56.2 82.0 96.0 113 119
10 t ha-1
7.2 10.4 14.5 17.6 25.9 37.4 41.2 93.6 117 121 129 134
Control Nil 3.5 11.4 11.8 12.5 18.0 21.8 23.0 26.0 34.5 42.0 59.3 76.8
Table 4C. Root length (cm) of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
3.4 17.4 22 24.2 34.5 38.1 41.2 42 48.6 51.2 55.3 59.2
7.5 t ha-1
9.3 22 24.8 25.2 28.6 35.5 53 56.4 62.7 65.3 64.7 72.1
10 t ha-1
11 12.9 19.4 21.2 29.1 29.8 52.5 61.8 63.7 68.9 80.1 83.4
Inorganic
fertilizer
5 t ha-1
8.2 11.9 18 19.2 20.2 22.7 24.2 29 36.1 47.4 44.8 55.7
7.5 t ha-1
7.4 13.5 19.6 20.4 31.2 38.1 52.5 69.2 79.6 79 85.3 89.1
10 t ha-1
9.6 12.1 14.2 23.6 25 28.4 43.2 43.2 55 57.2 65.8 50.1
Control Nil 3.8 13.7 14.0 23.1 29.1 29.2 37.4 42.0 45.7 47.5 49.3 50.2
128
Table 4D. Number of leaves of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
03 04 04 09 09 18 23 27 37 42 53 59
7.5 t ha-1
03 04 04 12 16 20 31 33 47 51 58 62
10 t ha-1
03 04 05 06 15 23 31 37 46 59 61 65
Inorganic
fertilizer
5 t ha-1
03 04 04 07 09 19 22 27 36 38 49 51
7.5 t ha-1
03 04 04 09 20 22 28 35 54 59 63 68
10 t ha-1
03 03 03 06 18 33 37 48 59 66 69 72
Control Nil 03 03 04 09 15 17 21 25 28 31 35 42
Table 4E. Number of nodules of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
Nil 5 7 14 15 18 20 21 18 21 27 28
7.5 t ha-1
Nil 7 11 14 19 25 28 31 28 34 41 39
10 t ha-1
Nil 2 7 9 12 19 30 35 33 39 37 45
Inorganic
fertilizer
5 t ha-1
Nil 3 7 9 12 21 21 33 24 27 26 38
7.5 t ha-1
Nil 3 9 12 15 20 20 22 31 29 38 37
10 t ha-1
Nil 4 8 9 15 17 19 29 28 22 37 34
Control Nil Nil 0 7 8 12 14 19 11 11 9 16 14
Table 4F. Shoot dry weight (g) of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.05 0.11 0.15 0.29 0.38 0.74 1.29 2.35 14.2 22.6 36.7 37.1
7.5 t ha-1
0.05 0.16 0.23 0.70 0.77 0.98 4.72 5.57 15.3 34.1 46.2 47.6
10 t ha-1
0.07 0.15 0.38 0.66 0.78 1.49 3.87 11.7 17.6 43.0 51.8 57.3
Inorganic
fertilizer
5 t ha-1
0.05 0.09 0.16 0.27 0.36 1.08 1.58 2.07 18.9 23.5 33.7 39.8
7.5 t ha-1
0.08 0.13 0.17 0.44 1.14 1.22 3.11 3.70 19.4 30.1 35.8 43.2
10 t ha-1
0.05 0.09 0.18 0.41 1.37 3.23 6.06 9.71 20.6 33.9 45.3 52.4
Control Nil 0.03 0.11 0.21 0.40 1.0 1.49 4.57 6.80 12.1 41.5 44.1 45.3
129
Table 4G. Root dry weight (g) of cluster bean at different weeks
Table 4H. Leaf chlorophyll a (mg g-1
) content of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.244 0.436 1.064 1.222 1.564 1.659 1.662 1.701 1.726 1.735 1.798 1.825
7.5 t ha-1
0.225 0.516 0.980 1.201 1.377 1.654 1.678 1.759 1.769 1.802 1.858 1.916
10 t ha-1
0.336 0.758 1.154 1.470 1.570 1.727 1.763 0.769 1.813 1.841 1.871 1.866
Inorganic
fertilizer
5 t ha-1
0.388 0.544 0.992 1.184 1.434 1.663 1.766 1.783 1.819 1.826 1.837 1.840
7.5 t ha-1
0.368 0.755 1.083 1.169 1.477 1.770 1.962 2.059 2.096 2.115 2.148 2.236
10 t ha-1
0.568 0.847 1.044 1.172 1.678 2.080 2.179 2.179 2.197 2.206 2.225 2.291
Control Nil 0.103 0.465 0.64 0.907 1.128 1.164 1.177 1.193 1.201 1.224 1.238 1.316
Table 4I. Leaf chlorophyll b (mg g-1
) content of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.113 0.311 0.623 0.971 1.097 1.375 1.390 1.421 1.475 1.519 1.590 1.613
7.5 t ha-1
0.242 0.310 0.827 0.752 1.032 1.293 1.369 1.401 1.460 1.593 1.616 1.629
10 t ha-1
0.210 0.517 0.970 1.111 1.152 1.440 1.463 1.790 1.875 1.826 1.989 2.051
Inorganic
fertilizer
5 t ha-1
0.223 0.613 0.735 0.827 1.073 1.189 1.164 1.181 1.325 1.419 1.487 1.513
7.5 t ha-1
0.120 0.318 1.069 1.166 1.532 1.795 1.882 1.903 2.016 2.054 2.102 2.117
10 t ha-1
0.218 0.412 1.012 1.124 1.799 1.921 2.156 2.166 2.185 2.204 2.217 2.279
Control Nil 0.032 0.155 0.223 0.504 1.612 1.935 1.065 1.409 1.510 1.532 1.544 1.451
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.01 0.02 0.02 0.04 0.14 0.23 0.43 0.97 1.87 2.93 4.41 4.78
7.5 t ha-1
0.01 0.03 0.03 0.07 0.20 0.34 0.87 2.02 2.55 4.81 6.28 6.82
10 t ha-1
0.01 0.02 0.04 0.05 0.18 0.39 0.69 1.85 3.29 4.02 6.52 7.22
Inorganic
fertilizer
5 t ha-1
0.01 0.01 0.02 0.03 0.23 0.21 0.42 1.65 3.05 4.50 6.60 7.53
7.5 t ha-1
0.01 0.02 0.05 0.07 0.29 0.37 0.76 2.27 3.81 5.70 8.21 9.39
10 t ha-1
0.01 0.03 0.04 0.11 0.28 1.13 1.54 4.04 5.87 7.42 10.2 12.7
Control Nil 0.01 0.01 0.02 0.03 0.10 0.26 0.29 0.98 1.76 2.93 3.16 3.21
130
Table 4J. Leaf total chlorophyll (mg g-1
) content of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.357 0.747 1.687 2.193 2.661 3.034 3.052 3.122 3.201 3.254 3.388 3.438
7.5 t ha-1
0.467 0.826 1.807 1.953 2.409 2.947 3.047 3.16 3.229 3.395 3.474 3.545
10 t ha-1
0.546 1.275 2.124 2.581 2.722 3.167 3.226 2.559 3.688 3.667 3.86 3.917
Inorganic
fertilizer
5 t ha-1
0.611 1.157 1.727 2.011 2.507 2.852 2.93 2.964 3.144 3.245 3.324 3.353
7.5 t ha-1
0.488 1.073 2.152 2.335 3.009 3.565 3.844 3.962 4.112 4.169 4.25 4.353
10 t ha-1
0.786 1.259 2.056 2.296 3.477 4.001 4.335 4.345 4.382 4.41 4.442 4.570
Control Nil 0.135 0.62 0.863 1.411 2.740 3.099 2.242 2.602 2.711 2.756 2.782 2.767
Table 4K. Leaf carotenoids (mg g-1
) content of cluster bean at different weeks
Treatment Amount Weeks
1 2 3 4 5 6 7 8 9 10 11 12
Vermicompost
5 t ha-1
0.119 0.121 0.145 0.209 0.300 0.205 0.214 0.221 0.236 0.258 0.271 0.293
7.5 t ha-1
0.108 0.154 0.228 0.259 0.313 0.334 0.345 0.396 0.418 0.437 0.441 0.462
10 t ha-1
0.123 0.133 0.331 0.291 0.326 0.347 0.324 0.390 0.428 0.459 0.462 0.459
Inorganic
fertilizer
5 t ha-1
0.132 0.156 0.182 0.190 0.213 0.231 0.259 0.268 0.281 0.314 0.340 0.358
7.5 t ha-1
0.140 0.171 0.184 0.215 0.253 0.280 0.298 0.311 0.336 0.348 0.352 0.361
10 t ha-1
0.175 0.197 0.203 0.222 0.288 0.294 0.312 0.322 0.342 0.348 0.356 0.374
Control Nil 0.102 0.122 0.136 0.158 0.164 0.191 0.203 0.218 0.224 0.236 0.248 0.261
EFFECT OF VERMICAST GENERATED FROM ALLELOPATHIC
WEEDS AND PAPER WASTE ON PHYSICAL AND CHEMICAL
PROPERTIES OF POTTING SOIL GROWING CLUSTER BEAN
(CYAMOPSIS TETRAGONOLOBA)
Chapter
9
131
A paper based on this chapter has been
submitted for publication.
CChhaapptteerr 99
Effect of vermicast generated from allelopathic weeds and
paper waste on physical and chemical properties of potting
soil growing cluster bean (Cyamopsis tetragonoloba)
Abstract
There is accumulation of scientific evidence on the positive impact of vermicompost on plant growth.
However, the vermicompost derived from different parent materials have different physical, chemical and
biological qualities, and their impact on plant growth is also reported to vary considerably. To understand the
factors which attribute the differential impact of vermicompost from different parent materials on the growth
and yield of plants, it is necessary to recognize their impact on soil health. Hence, in the present study, the
application of vermicast generated from different organic wastes such as paper waste, leaves of ipomoea
(Ipomoea carnea), and of lantana (Lantana camara) on physical and chemical properties of potting soil
housing cluster bean (Cyamopsis tetragonoloba) were investigated. Seeds of cluster bean were sown in soil
to which vermicast was applied at the rate of 5, 7.5 and 10 t ha-1
respectively. In another set of treatment,
amounts of essential nutrients equivalent to those present in the vermicast treatments were applied in the
inorganic form and a control set comprising of only soil has been kept without any nutrient supplement.
Samples from all these treatments were collected on weekly basis during different stages of plant growth.
The essential physical and chemical variables of soil which is related to the plant growth were studied. The
results reveal that vermicast application created a suitable physical environment and also reduced the nutrient
loss at surface soil than the inorganic fertilizer.
1. Introduction
Vermicompost has long been recognized as
beneficial organic soil amendment in agriculture for
the maintenance of soil fertility to support plant
growth. Supplement of vermicompost, stimulate
seed germination, vegetative growth, shoot and root
development of several plant species. Stimulation
of flowering, increase in fruit yield and its
nutritional quality are also reported with
vermicompost supplement (Edwards et al., 2011).
Additionally, it increases the disease resistance of
plants to parasitic nematodes, insects, and mites
(Yardim et al., 2006; Zaller, 2006; Edwards et al.,
2009; Simsek-Ersahin et al., 2009; Simsek-Ersahin,
2011). The improvement in plant production and
protection with vermicompost substitution could be
attributed to several factors. First, vermicompost
amendments improve the overall physico-chemical
and biological characteristics of the soils that favor
better plant growth. The presence of plant growth-
influencing substances, such as plant growth
regulating hormones and humic acids in
vermicompost has also been suggested as a possible
factor contributing to increased plant growth and
yields (Tomati et al., 1988; Muscolo et al., 1999;
Arancon et al., 2003a; Arancon et al., 2006).
Through the vermicomposting technology,
wide range of organic wastes can be processed,
which includes paper waste (Elvira et al., 1997,
1998; Gajalakshmi and Abbasi, 2004), sewage
sludge (Domínguez et al., 2000; Khwairakpam and
Bhargava, 2009; Hait and Tare, 2011), urban
132
residues, and food and animal waste (Edwards et
al., 1985; Edwards, 1988; Domínguez and
Edwards, 1997; Atiyeh et al., 2001; Aira et al.,
2006; Garg et al., 2006; Suthar, 2007; Lazcano et
al., 2008), as well as horticultural residues
(Gajalakshmi et al., 2005; Gupta et al., 2007;
Pramanik et al., 2007; Suthar, 2007) and food
industry waste (Edwards, 1983; Butt, 1993;
Nogales et al., 1999a, 1999b, 2005). Nevertheless,
the vermicompost prepared from different organic
wastes, with different processes and duration,
produce a final product which differs in its quality
(Tandon, 1995; Campitelli and Ceppi, 2008).
The previous studies (Chapters 6, 7 and 8)
dealing with the effect of vermicast generated from
paper waste and from pernicious weed, lantana
(Lantana camara) and ipomoea (Ipomoea carnea)
on the germination, growth and yield of cluster
bean also manifests this finding. The vermicompost
generated from different parent material show
different physical and chemical qualities and their
influence on germination, growth and yield of test
plant also varied considerably. It was seen that the
application vermicompost derived from phytomass
such as lantana and ipomoea supported the
germination, growth and better fruit yield than
inorganic fertilizers. In contrast, the plants treated
with paper waste vermicompost exhibited lower
growth and yield than the equivalent inorganic
fertilizer. To understand the factors which attribute
the disparity on the growth and yields of plants
which were treated with vermicompost from
different parent materials, it is necessary to
understand their impact on soil health. Hence, the
objectives of the presence study are, (i) to evaluate
the impact of vermicast from different parent
materials on the physical and chemical properties of
potting soil germinating and growing cluster bean,
(ii) to determine the dose dependent changes in the
soil properties with vermicast application, and (iii)
to compare the impact of different vermicompost
on soil properties with soil fortified with inorganic
fertilizers (IF) in concentrations equivalent to those
present in the respective vermicompost (VC)
treatments.
2. Materials and methods
The studies were conducted at Pondicherry
University, Puducherry, India, located on the east
coast of the Indian peninsula (11°56’N, 79°53’E).
This region experiences hot summers during March
– July (maximum day temperature 35-38°C), and
mild winters during December - February
(maximum day temperature 29-32°C). The average
annual rainfall is about 1300 mm, concentrated
mainly during October – December but with a few
rainy days occurring in July–August and January as
well. For vermicomposting, reactors fabricated with
aluminum sheet of 140 liter volume were employed
for direct vermicomposting of lantana and ipomoea
leaves by a process recently developed by the
author’s group (Gajalakshmi et al., 2002, 2005;
Abbasi et al., 2009, 2011; Kumar et al., 2012;
Tauseef et al., 2013). As paper waste is almost
entirely cellulosic, with only traces of elements
other than C, H, and O, the feed was spiked with
9% w/w of cow dung in order to provide NPK and
other nutrients in adequate amounts and
vermicomposted in similar fashion (Gajalakshmi et
al., 2012). The periodically harvested vermicast
was stored in sealed plastic containers. The study
on germination and growth were conducted
outdoors with 49 liter containers (40 cm height with
surface area 35 x 35 cm), lined with high-density
polyethylene (HDPE) sheets. The soil used in the
experiments was collected from a previously
uncultivated piece of land so that the results are not
influenced by any earlier fertilizer application. It is
sandy loam soil which is known to be low on
organic carbon and nutrients. The physico-chemical
properties of the vermicast and of the soil with
which it was used are given in Table 1. The
experiments were carried out during July – October
which is reported to be a season ideal for growing
cluster beans in the study area (ICAR, 2011).
133
Table 1. Chemical and physical properties of vermicast and soil used in the study.
BDL – Below detection limit
For studying the impact of vermicast from
paper waste and leaves of lantana and ipomoea on
potting soil housing cluster bean, three sets of
experiments were conducted for each substrate. In
set I, one batch of containers was supplied with 7.5
t ha-1
of VC while two other batches were given 5
and 10 t ha-1
of VC. Set II comprised of chemical
fertilizers (IF) which were treated with nutrients N,
P, K, Ca, Mg, S, Fe, Mn, Cu, Zn, B, Mo and Cl in
concentrations equivalent to those present in the
respective vermicompost treatments. The last set
had no supplementation of nutrients and served as
control. Seven set, each comprising of 36
containers, or 252 containers in all, were prepared
in this fashion. The Pusanavabahar variety seeds of
the cluster bean, which is locally available in the
authors’ study area, was sown in all the containers.
Following germination, the plants were grown for
three months. Throughout the experiments,
adequate watering was done. Deweeding was done
periodically.
The soil samples from all the treatments were
collected on weekly basis during different stage of
plant growth, for physical and chemical analysis.
Undisturbed soil cores (5.7 cm diameter and 40 cm
Variables Vermicast
Lantana camara Ipomoea carnea Paper waste Soil
Chemical properties
pH 6.47±0.01 6.51±0.06 7.83±0.05 6.30±0.10
Total organic carbon g kg-1
330.3±4.3 356.8±12.1 259.6±8.2 8.87±0.02
Total Nitrogen g kg-1
18.4±0.1 19.8±0.2 11.7±0.4 2.66±0.02
Plant available form of
Phosphorus mg kg -1
80.7±0.9 37.83±0.60 71.40±3.31 0.41±0.01
Potassium g kg-1
5.73±0.03 2.12±0.04 1.98±0.10 0.40±0.00
Sulphur mg 100g-1
1.34±0.01 4.64±0.09 0.40±0.04 0.54±0.01
Calcium g kg-1
5.49±0.02 15.5±2.6 15.9±1.4 8.27±0.01
Magnesium g kg-1
5.77±0.05 4.85±0.09 7.13±0.07 0.09±0.02
Boron mg kg-1
46.2±0.8 78.1±2.3 7.28±0.09 26.9±1.2
Copper mg kg-1
44.2±3.9 10.6±0.3 16.9±1.0 5.08±0.15
Iron mg kg-1
50.6±2.0 270.3±14.7 208.3±11.4 59.9±2.9
Manganese mg kg-1
227.2±8.4 117.4±5.0 63.8±1.8 45.1±2.2
Zinc mg kg-1
162.4±6.0 107.0±7.7 94.1±2.2 55.0±2.6
Molybdenum mg kg-1
BDL BDL BDL BDL
Physical properties
Dry weight % 43.1±0.3 48.4±1.1 49.6±1.9 94.7±0.1
Bulk density g cm-3
0.40±0.00 0.26±0.02 0.24±0.01 1.28±0.00
Particle density g cm-3
1.35±0.01 1.30±0.05 1.21±0.03 2.70±0.12
Water-holding capacity % 248.0±10.2 261.9±16.6 118.0±11.2 36.9±2.4
Electrical conductivity mmhos cm-1
9.36±0.01 6.25±0.07 2.96±0.04 0.12±0.02
Total porosity % 70.6±0.2 80.2±1.8 80.08±1.0 53.1±2.0
Air filled porosity % 48.1±0.1 66.8±2.6 68.14±1.2 46.5±2.1
134
height) were taken from the 0 to 30 cm layer by
inserting core sampler gently into the soil. The
sampled cores were divided into two pieces (0-15
cm and 15-30 cm) and packed separately in airtight
polyethylene bags. These samples were stored at
2°C until analyses could take place. Forty two soil
samples per week (7 treatments x 3 replications x 2
soil depth layers: 0–15 cm, 15–30 cm), leading to
504 samples overall were collected during the
course of three months of the experiment. The
samples were air-dried at room temperature,
crushed, and passed through a 2 mm diameter
sieve, before analyses. For bulk density
measurement, undisturbed soil cores were collected
separately from each soil depth.
Total organic carbon (Corg) was measured by
modified dichromate redox method according to
Heanes (1984). Total nitrogen (Ntot) content was
determined by the modified Kjeldahl method
(Kandeler, 1993) using Kel Plus™ semi-automated
digester and distillation units. Inorganic N - NH4+
and NO3- were extracted in 2M KCl solution (1:10,
w/v) and determined by modified indophenol blue
and Devarda’s alloy methods, respectively (Jones
2001; Bashour and Sayegh, 2007). Extractable
potassium (Kext) and calcium (Caext) were
determined using a flame photometer (Elico™
CL378) after extraction with neutral 1N ammonium
acetate solution (Carter and Gregorich, 2008).
Extractable form of magnesium (Mgext), boron
(Bext), copper (Cuext), iron (Feext), manganese
(Mnext), zinc (Znext), molybdenum (Moext) were
determined using a Jobin Yvon – Ultima 2 model
inductively coupled plasma atomic emission
spectroscopy (ICP-AES) by extracting
sample/solution ratio of 1:25 (w/v) with Mehlich 3
extraction solution (Mehlich, 1984). The same
extract was used to determine the extractable
phosphorus (Pext) according to the ammonium
molybdate-ascorbic acid method (Knudsen and
Beegle, 1988).
The pH and electrical conductivity (EC) of
the samples were measured in suspension of 1:2
(v/w) by using EI™ 611E EC meter and Digison™
digital pH meter 7007 respectively. Bulk density
and particle density were measured by undisturbed
core and volumetric flask methods, respectively
(Bashour and Sayegh, 2007). The total and water-
filled porosity (WFPS) were calculated from the
particle and bulk density values of respective
samples using standard formulae (Carter and
Gregorich, 2008). Water-holding capacity (WHC)
of the samples was obtained by determining their
water retention ability after they were immersed in
water and the excess water was drained off
(Margesin and Schinner, 2005).
One-way ANOVA and post hoc LSD tests
were employed for determining significance of
difference between the results. SPSS version 16
was used for this purpose.
3. Results and discussion
3.1. Physical properties
The physical properties of soil were
significantly affected by different VC and IF
treatments (p<0.05; Tables 2-8). The soil treated
with VC from all three substrates had significantly
lower bulk density than the ones treated with
corresponding IF treatments and control (Table 2).
During the initial month, there was no significant
difference in bulk density of soils treated with VC
and the equivalent IF. However, in the second and
third months, the bulk density of soil treated with
ipomoea based VC was about 7% lower than the IF
treatments and those of lantana and paper waste VC
showed 8 and 12% lesser bulk density, respectively.
The bulk density of lantana based VC treated soil
were 23% lower than the control; whereas the
ipomoea and paper waste VC treatments showed 20
and 16% lesser bulk density, respectively. In all the
VC treatments, increasing dose of VC showed a
steady decline in bulk density, which was possibly
due to the reasons, (i) addition of lower density
organic matter in the form of vermicast might have
diluted the bulk density of mineral soil (Tejada et
135
Table 2. Changes in the bulk density (g cm−3
) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or equivalent
inorganic fertilizers (IF), at different periods of time (mean ± SD).
Treatments Lantana Ipomoea Paper waste
July Aug Sep July Aug Sep July Aug Sep
Layer
0-15 cm VC 1.08±0.06 1.13±0.04 1.15±0.04 1.06±0.11 1.14±0.05 1.19±0.05 1.11±0.05 1.17±0.04 1.23±0.03
IF 1.07±0.07 1.18±0.04 1.25±0.03 1.10±0.08 1.22±0.06 1.27±0.04 1.10±0.05 1.17±0.03 1.21±0.02
Cont. 1.23±0.04 1.27±0.01 1.29±0.01 1.26±0.04 1.30±0.01 1.32±0.01 1.26±0.04 1.30±0.01 1.32±0.01
15-30 cm VC 1.05±0.06 1.10±0.05 1.13±0.05 1.03±0.07 1.12±0.06 1.17±0.05 1.08±0.04 1.15±0.04 1.22±0.03
IF 1.06±0.08 1.16±0.04 1.23±0.03 1.07±0.08 1.18±0.05 1.24±0.05 1.09±0.06 1.16±0.04 1.20±0.02
Cont. 1.20±0.03 1.23±0.01 1.27±0.03 1.23±0.03 1.26±0.01 1.30±0.03 1.23±0.03 1.26±0.01 1.30±0.03
F value 6.517***
22.59***
3.847**
5.248**
12.22***
16.49***
9.309***
17.44***
25.04***
Amount
5 t ha-1
VC 1.07±0.06 1.15±0.01 1.18±0.01 1.13±0.02 1.18±0.03 1.25±0.02 1.14±0.04 1.20±0.02 1.25±0.03
IF 1.06±0.05 1.19±0.03 1.26±0.02 1.15±0.08 1.27±0.03 1.31±0.01 1.07±0.05 1.16±0.01 1.21±0.01
7.5 t ha-1
VC 1.12±0.02 1.14±0.01 1.16±0.01 0.94±0.05 1.08±0.05 1.15±0.03 1.09±0.02 1.16±0.02 1.22±0.02
IF 0.99±0.05 1.13±0.04 1.21±0.02 1.01±0.06 1.15±0.02 1.22±0.04 1.07±0.03 1.14±0.03 1.19±0.01
10 t ha-1
VC 1.01±0.04 1.05±0.02 1.08±0.02 1.06±0.07 1.13±0.01 1.14±0.01 1.05±0.03 1.12±0.02 1.20±0.03
IF 1.14±0.03 1.20±0.03 1.25±0.03 1.09±0.03 1.19±0.03 1.24±0.02 1.15±0.03 1.19±0.01 1.23±0.01
Control
1.21±0.04 1.25±0.03 1.28±0.02 1.24±0.04 1.28±0.03 1.31±0.02 1.24±0.04 1.28±0.03 1.31±0.02
F value
21.09***
68.46***
4.212**
20.50***
45.52***
82.83***
21.99***
47.96***
44.18***
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
136
Table 3. Changes in the particle density (g cm−3
) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or equivalent
inorganic fertilizers (IF), at different periods of time (mean ± SD).
Treatments Lantana Ipomoea Paper waste
July Aug Sep July Aug Sep July Aug Sep
Layer
0-15 cm VC 2.63±0.03 2.64±0.07 2.70±0.30 2.65±0.04 2.69±0.04 2.68±0.05 2.61±0.04 2.62±0.05 2.65±0.04
IF 2.65±0.09 2.65±0.02 2.63±0.03 2.66±0.05 2.66±0.03 2.70±0.08 2.65±0.03 2.67±0.06 2.66±0.02
Cont. 2.64±0.02 2.65±0.05 2.66±0.03 2.64±0.02 2.65±0.05 2.66±0.03 2.64±0.02 2.65±0.05 2.66±0.03
15-30 cm VC 2.62±0.04 2.64±0.04 2.67±0.04 2.63±0.02 2.65±0.03 2.65±0.03 2.63±0.05 2.63±0.07 2.63±0.05
IF 2.64±0.05 2.65±0.03 2.92±1.03 2.67±0.08 2.65±0.03 2.64±0.08 2.66±0.02 2.66±0.04 2.65±0.04
Cont. 2.64±0.02 2.68±0.04 2.64±0.03 2.64±0.02 2.68±0.04 2.64±0.03 2.64±0.02 2.68±0.04 2.64±0.03
F value 0.401n.s
0.748n.s
0.637n.s
0.597n.s
1.990n.s
1.738n.s
3.183* 1.589
n.s 0.998
n.s
Amount
5 t ha-1 VC 2.62±0.03 2.62±0.07 2.61±0.11 2.63±0.03 2.67±0.02 2.67±0.05 2.58±0.04 2.63±0.08 2.60±0.04
IF 2.66±0.05 2.64±0.02 2.63±0.03 2.68±0.04 2.67±0.04 2.67±0.05 2.66±0.01 2.70±0.04 2.67±0.03
7.5 t ha-1 VC 2.63±0.02 2.65±0.04 2.75±0.34 2.65±0.04 2.66±0.05 2.65±0.06 2.62±0.03 2.61±0.03 2.66±0.04
IF 2.60±0.05 2.65±0.03 2.66±0.04 2.68±0.10 2.64±0.02 2.67±0.07 2.66±0.03 2.64±0.06 2.64±0.02
10 t ha-1 VC 2.61±0.04 2.65±0.04 2.68±0.05 2.64±0.03 2.68±0.04 2.66±0.04 2.65±0.04 2.64±0.06 2.66±0.03
IF 2.67±0.08 2.66±0.04 3.04±1.26 2.64±0.04 2.64±0.03 2.67±0.12 2.65±0.02 2.66±0.04 2.65±0.03
Control
2.64±0.02 2.67±0.05 2.65±0.03 2.64±0.02 2.67±0.05 2.65±0.03 2.64±0.02 2.67±0.05 2.65±0.03
F value
1.852n.s
1.149n.s
0.891n.s
0.924n.s
1.228n.s
0.190n.s
5.736***
2.363* 5.185
***
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
137
Table 4. Changes in the total porosity (%) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or equivalent
inorganic fertilizers (IF), at different periods of time (mean ± SD).
Treatments Lantana Ipomoea Paper waste
July Aug Sep July Aug Sep July Aug Sep
Layer
0-15 cm VC 59.0±2.2 57.2±2.3 56.9±4.1 60.0±4.5 57.4±2.0 55.5±2.1 57.6±2.4 55.3±2.1 53.5±1.6
IF 59.6±1.9 55.4±1.8 52.7±1.7 58.8±2.9 54.2±1.8 52.9±2.1 58.5±2.0 56.2±1.2 54.5±0.7
Cont. 53.6±1.6 52.1±1.0 51.4±0.6 52.5±1.6 50.9±1.0 50.3±0.6 52.5±1.6 50.9±1.0 50.3±0.6
15-30 cm VC 59.9±2.2 58.2±1.9 57.7±2.3 60.9±2.6 58.0±2.1 55.8±2.0 58.8±2.0 56.3±1.8 53.8±1.8
IF 60.0±3.2 56.1±1.4 55.3±7.2 59.8±3.5 55.4±2.0 53.1±2.3 59.2±2.3 56.5±1.5 54.6±1.0
Cont. 54.7±1.1 54.2±1.0 51.9±1.4 53.6±1.1 53.1±1.0 50.7±1.4 53.6±1.1 53.1±1.0 50.7±1.4
F value 7.339***
15.83***
2.042n.s
4.807**
13.63***
11.51***
7.605***
10.33***
14.56***
Amount
5 t ha-1
VC 59.1±1.9 56.0±1.5 54.8±2.2 57.1±1.3 55.6±1.2 53.2±1.2 55.8±1.6 54.1±1.7 51.9±1.3
IF 60.3±2.1 55.0±1.0 52.1±1.1 57.2±2.7 52.6±1.1 51.1±0.9 59.9±2.0 57.1±0.4 54.8±0.7
7.5 t ha-1
VC 57.7±0.8 56.9±0.6 57.4±3.9 64.5±2.0 59.6±1.6 56.7±1.0 58.3±0.6 55.7±0.7 54.1±1.1
IF 61.8±2.0 57.2±1.6 54.5±1.3 62.2±3.0 56.5±1.0 54.4±1.9 59.9±1.3 56.8±1.6 55.1±0.6
10 t ha-1
VC 61.5±1.6 60.3±0.6 59.7±1.4 59.7±2.4 58.0±0.8 57.1±0.5 60.4±1.3 57.7±1.4 55.0±1.0
IF 57.3±1.0 55.0±1.4 55.4±9.1 58.5±1.1 55.2±1.0 53.6±2.1 56.7±1.1 55.2±0.8 53.7±0.5
Control
54.2±1.4 53.1±1.4 51.6±1.0 53.1±1.4 52.0±1.5 50.5±1.0 53.1±1.4 52.0±1.5 50.5±1.0
F value
19.01***
37.78***
2.330* 19.21
*** 41.34
*** 34.38
*** 23.06
*** 20.25
*** 36.52
***
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
138
Table 5. Changes in the water-filled pore space (%) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or
equivalent inorganic fertilizers (IF), at different periods of time (mean ± SD).
Treatments Lantana Ipomoea Paper waste
July Aug Sep July Aug Sep July Aug Sep
Layer
0-15 cm VC 24.7±4.7 25.3±5.3 30.2±4.3 20.0±5.4 26.4±3.5 27.4±3.9 24.0±3.4 26.0±4.2 28.9±4.7
IF 23.3±2.5 27.3±5.5 31.6±7.6 25.4±6.2 29.8±6.0 29.1±5.8 22.9±4.5 25.7±4.9 27.6±3.9
Cont. 26.1±1.7 28.5±5.2 29.1±3.5 27.3±1.8 29.9±5.5 30.5±3.7 27.3±1.8 29.9±5.5 30.5±3.7
15-30 cm VC 26.8±3.7 30.5±4.3 29.2±4.6 21.2±5.1 25.9±4.3 32.4±5.9 27.2±3.9 29.5±4.7 33.0±4.8
IF 27.6±3.0 31.2±5.3 33.7±5.0 25.8±4.9 30.7±6.5 33.5±5.0 25.9±3.3 28.4±4.2 31.1±3.6
Cont. 29.2±1.2 30.6±3.2 33.0±2.1 30.5±1.2 32.1±3.3 34.5±2.3 30.5±1.2 32.1±3.3 34.5±2.3
F value 2.891* 2.865
* 2.103
n.s 3.168
* 2.060
n.s 3.542
** 2.987
* 2.115
n.s 4.019
**
Amount
5 t ha-1
VC 25.9±5.1 30.2±4.6 31.3±4.2 24.7±2.9 26.4±4.7 31.8±5.8 26.6±5.3 30.9±4.1 33.3±5.1
IF 24.7±3.2 31.5±7.5 37.0±7.2 27.1±4.8 33.3±6.5 32.9±4.9 24.1±5.0 24.4±4.7 30.3±4.1
7.5 t ha-1
VC 27.3±3.7 30.1±3.4 29.5±5.8 16.0±2.4 24.5±3.4 28.5±5.7 25.6±3.2 27.7±4.4 31.2±5.6
IF 24.7±3.3 26.5±4.2 30.6±5.3 20.4±2.2 25.1±4.0 30.5±5.1 22.8±3.1 28.6±2.7 27.6±4.3
10 t ha-1
VC 24.1±4.0 23.4±5.5 28.3±2.4 21.2±5.4 27.5±3.3 29.5±5.0 24.5±3.3 24.7±3.7 28.3±3.4
IF 27.1±3.9 29.8±3.8 30.4±4.6 29.4±4.5 32.3±4.4 30.6±7.3 26.2±4.0 28.1±5.5 30.2±3.8
Control
27.6±2.1 29.6±4.1 31.1±3.4 28.9±2.2 31.0±4.3 32.5±3.6 28.9±2.2 31.0±4.3 32.5±3.6
F value
1.156n.s
3.053* 1.073
n.s 10.87
*** 5.069
*** 0.880
n.s 1.565
n.s 3.046
* 2.302
*
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
139
Table 6. Changes in the water holding capacity (%) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or
equivalent inorganic fertilizers (IF), at different periods of time (mean ± SD).
Treatments Lantana Ipomoea Paper waste
July Aug Sep July Aug Sep July Aug Sep
Layer
0-15 cm VC 41.1±4.2 43.9±3.5 46.4±3.2 45.4±5.1 48.6±6.0 50.6±6.6 42.2±5.0 45.1±5.0 48.1±5.6
IF 33.5±1.6 35.3±0.9 36.4±0.9 35.2±2.8 38.6±2.8 39.7±2.6 36.5±1.6 39.3±3.0 42.6±2.3
Cont. 31.0±0.6 33.4±0.6 35.5±0.8 31.0±0.6 33.4±0.6 35.5±0.8 31.0±0.6 33.4±0.6 35.5±0.8
15-30 cm VC 39.0±2.6 41.7±2.5 44.3±2.6 43.3±5.3 46.6±6.0 49.8±6.3 39.2±4.4 42.1±4.1 44.9±4.0
IF 32.1±1.5 33.3±0.9 34.5±0.7 35.0±1.6 37.5±1.3 39.9±1.1 35.3±1.3 37.7±1.3 40.5±1.8
Cont. 32.9±0.2 33.8±1.0 36.3±0.7 32.9±0.2 33.8±1.0 36.3±0.7 32.9±0.2 33.8±1.0 36.3±0.7
F value 18.87***
46.53***
83.67***
15.45***
18.71***
23.31***
8.784***
12.99***
16.94***
Amount
5 t ha-1
VC 36.5±0.8 39.5±1.7 41.7±1.1 38.3±1.0 39.8±1.2 41.5±0.5 34.9±1.3 37.7±1.5 40.1±1.0
IF 32.6±1.7 34.1±1.6 35.4±1.2 36.4±1.1 38.7±1.9 40.4±0.8 34.8±0.7 36.7±0.9 39.6±1.4
7.5 t ha-1
VC 39.9±1.8 42.8±1.1 46.3±1.1 45.0±2.7 50.3±2.5 53.5±0.7 42.0±2.3 45.5±2.0 49.1±2.7
IF 34.3±1.2 35.1±1.3 36.2±1.3 36.3±1.2 39.1±2.2 41.2±1.3 37.1±1.2 39.4±2.2 42.8±0.7
10 t ha-1
VC 43.8±2.6 46.1±2.2 48.1±1.9 49.7±2.2 52.7±1.5 55.5±1.6 45.2±2.3 47.5±2.3 50.2±2.3
IF 31.5±0.7 33.9±1.1 34.9±1.0 32.6±1.6 36.3±1.8 37.8±1.7 35.8±1.6 39.4±2.8 42.3±2.7
Control
31.9±1.1 33.6±0.8 35.9±0.8 31.9±1.1 33.6±0.8 35.9±0.8 31.9±1.1 33.6±0.8 35.9±0.8
F value
54.93***
95.63***
207.6***
91.47***
132.8***
443.9***
51.31***
51.92***
78.60***
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
140
Table 7. Changes in the electrical conductivity (mmhos cm−1
) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste,
or equivalent inorganic fertilizers (IF), at different periods of time (mean ± SD).
Treatments Lantana Ipomoea Paper waste
July Aug Sep July Aug Sep July Aug Sep
Layer
0-15 cm VC 0.18±0.03 0.20±0.02 0.19±0.02 0.17±0.02 0.18±0.02 0.18±0.01 0.15±0.02 0.16±0.02 0.15±0.02
IF 0.19±0.02 0.19±0.02 0.17±0.02 0.19±0.02 0.19±0.02 0.17±0.02 0.17±0.02 0.17±0.01 0.16±0.00
Cont. 0.11±0.00 0.12±0.00 0.12±0.00 0.11±0.00 0.12±0.00 0.12±0.00 0.11±0.00 0.12±0.00 0.12±0.00
15-30 cm VC 0.18±0.01 0.19±0.02 0.18±0.01 0.17±0.01 0.18±0.01 0.18±0.01 0.14±0.01 0.14±0.02 0.13±0.02
IF 0.20±0.03 0.20±0.03 0.18±0.02 0.20±0.03 0.19±0.02 0.18±0.02 0.18±0.02 0.18±0.02 0.17±0.02
Cont. 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00
F value 15.59***
21.57***
32.74***
23.29***
28.97***
50.93***
20.46***
19.33***
29.00***
Amount
5 t ha-1
VC 0.16±0.00 0.17±0.00 0.17±0.00 0.16±0.01 0.17±0.00 0.17±0.00 0.14±0.01 0.14±0.01 0.12±0.01
IF 0.17±0.01 0.16±0.01 0.15±0.01 0.17±0.01 0.16±0.00 0.16±0.01 0.17±0.01 0.17±0.01 0.16±0.00
7.5 t ha-1
VC 0.17±0.01 0.20±0.01 0.19±0.01 0.17±0.01 0.19±0.01 0.19±0.00 0.14±0.01 0.14±0.01 0.13±0.01
IF 0.20±0.01 0.20±0.01 0.18±0.01 0.20±0.01 0.20±0.01 0.18±0.01 0.16±0.01 0.16±0.01 0.16±0.00
10 t ha-1
VC 0.20±0.02 0.21±0.01 0.20±0.01 0.18±0.01 0.19±0.01 0.19±0.00 0.17±0.01 0.18±0.01 0.16±0.01
IF 0.22±0.01 0.22±0.01 0.20±0.00 0.22±0.01 0.21±0.01 0.19±0.01 0.19±0.01 0.20±0.02 0.18±0.02
Control
0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00
F value
58.90***
102.3***
227.6***
77.90***
184.5***
343.5***
62.71***
53.14***
56.16***
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
141
Table 8. Changes in the pH of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or equivalent inorganic fertilizers
(IF), at different periods of time (mean ± SD).
Treatments
Lantana Ipomoea Paper waste
July Aug Sep July Aug Sep July Aug Sep
Layer
0-15 cm VC 6.22±0.07 6.17±0.09 6.15±0.11 6.30±0.01 6.31±0.03 6.30±0.03 6.40±0.04 6.70±0.28 6.82±0.29
IF 6.24±0.06 6.22±0.06 6.21±0.06 6.52±0.29 6.61±0.41 6.61±0.39 6.83±0.53 7.03±0.48 7.07±0.46
Cont. 6.30±0.00 6.28±0.01 6.27±0.00 6.30±0.00 6.28±0.01 6.27±0.00 6.30±0.00 6.28±0.01 6.27±0.00
15-30 cm VC 6.06±0.08 5.93±0.29 5.98±0.01 6.10±0.13 6.08±0.13 6.07±0.13 6.28±0.08 6.33±0.08 6.36±0.08
IF 6.09±0.08 6.04±0.05 6.02±0.04 6.16±0.10 6.17±0.12 6.18±0.13 6.34±0.08 6.40±0.09 6.42±0.08
Cont. 6.27±0.02 6.21±0.01 6.19±0.01 6.27±0.02 6.21±0.01 6.19±0.01 6.27±0.02 6.21±0.01 6.19±0.01
F value 15.57***
9.140***
41.59***
9.745***
0.901n.s
14.71***
7.187***
17.03***
1.199n.s
Amount
5 t ha-1
VC 6.11±0.14 6.09±0.14 6.09±0.13 6.28±0.02 6.28±0.02 6.27±0.02 6.29±0.09 6.36±0.13 6.40±0.14
IF 6.19±0.12 6.17±0.12 6.16±0.12 6.28±0.03 6.27±0.03 6.27±0.02 6.34±0.07 6.41±0.11 6.44±0.11
7.5 t ha-1
VC 6.22±0.08 6.01±0.41 6.11±0.13 6.16±0.15 6.14±0.15 6.13±0.15 6.33±0.11 6.45±0.14 6.54±0.20
IF 6.22±0.06 6.17±0.08 6.14±0.10 6.19±0.18 6.19±0.20 6.20±0.20 6.45±0.15 6.68±0.33 6.72±0.34
10 t ha-1
VC 6.10±0.06 6.05±0.03 5.99±0.02 6.16±0.17 6.16±0.19 6.16±0.19 6.40±0.03 6.74±0.35 6.83±0.40
IF 6.08±0.09 6.06±0.09 6.05±0.09 6.55±0.40 6.71±0.48 6.72±0.45 6.97±0.61 7.06±0.59 7.08±0.58
Control
6.28±0.02 6.24±0.04 6.23±0.04 6.28±0.02 6.24±0.04 6.23±0.04 6.28±0.02 6.24±0.04 6.23±0.04
F value
4.572**
1.754n.s
6.263***
3.162* 0.997
n.s 9.221
*** 5.883
*** 7.027
*** 1.044
n.s
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
142
al., 2009), and (ii) vermicast containing binding
agents such as polysaccharides and bacterial gums
would have promoted the soil aggregation, which
might have increased the porosity of soil resulting
in reduction in bulk density (Bhatia and Shukla,
1982; Rasool et al., 2007). In the case of IF treated
soil, the highest reduction in bulk density was
recorded in 7.5 t ha-1
VC equivalent IF treatments,
followed by 10 and 5 t ha-1
. In most of the cases,
bulk density of the IF treated soil were 10% lesser
than the control. Few past studies also showed that
the balanced inorganic fertilizer application
increased the porosity of soil, resulting in the
reduction of bulk density (Yeoh and Oades, 1981;
Schjonning et al., 1994; Prasad and Sinha, 2000).
In all the treatments, the first 15 cm soil layer had
higher bulk density than deeper soil layer (15-30
cm). The particle density of untreated soil was
2.70±0.12 g cm-3
, which has not changed
considerably throughout the experiment (Table 3).
Total porosity of ipomoea and lantana based
VC treated soil were about 5% higher than the
corresponding IF treatments; whereas the paper
waste VC treated soil had shown 2% lower porosity
than the IF treatments (Table 4). Lantana and
ipomoea VC treated soil showed about 10% higher
porosity compared to the control treatment. Paper
waste based VC treatments increased the porosity
to the extent of 6%. In all IF treatments, porosity of
soil increased to the extent of 7% in comparison
with control. The VC and balanced fertilizer
application might have improved the aggregation
by microbial extracellular polysaccharide
production, filamentous structures (e.g. fungal
hyphae) and extruded biopolymer-induced
aggregation (Oades, 1993), resulting in an increase
in porosity of soil (Bhatia and Shukla, 1982).
Moreover, the VC amendment may have enhanced
the microbial decomposition of organic matter it
contained, and subsequent gaseous release fractured
the soil matrix which might have increased the pore
space (Pagliai et al., 1981).
The water-filled pore space (WFPS) of soils
has not showed any significant variation with
different treatments (Table 5). In most of the cases,
WFPS of soil treated with VC and IF were similar
to the control. The different doses of VC and IF
applications have also not had any significant
difference in WFPS. The water-holding capacity
(WHC) of soil significantly increased with
increasing dose of VC application (Table 6). Higher
WHC of VC treated soils may be probably the
reason of high pore space in these treatments (Liu
et al., 2013). Amongst different VC treatments, the
ipomoea VC showed higher increase of about 28%
in WHC compared to the control. In both lantana
and paper waste VC; it increased to the extent of
21%. The IF treatments increased the WHC to the
extent of 10% in comparison to the control. In all
the treatments, increase in WHC was prominent in
the first 15 cm layer than the soil below. EC of both
VC and IF treated soil were higher than the control
(Table 7). In the lantana and ipomoea based VC
and equivalent IF treatments, EC increased to the
extent of 39%, and in the paper waste VC and
equivalent IF treatments, it increased to the extent
of 15 and 31%, respectively in comparison to the
control. During the first month, higher EC was
recorded in both VC and IF treatments, and then
declined till the end. There was no significant
difference in EC of soil either treated with different
VC and IF, or collected from different layers. The
pH of soil from all the treatments was in the range
of 5.95 – 7.64. All IF treatments showed slightly
higher pH than the corresponding VC treatments
(Table 8).
3.2. Chemical properties
Chemical characteristics of soil were
significantly influenced by different treatments
(p<0.001; Tables 9-19). Application of VC
significantly increased the organic carbon (Corg)
content in soil; more than 50% higher compared to
the control and IF treatments during the first month
(Table 9). Among different treatments, the lantana
143
based VC increased the Corg maximally, followed
by ipomoea and paper waste VC. The Corg content
of IF treated soil did not show any significant
difference compared to the control. However,
throughout the experiment the Corg in all the
treatments increased, which may be due to the
addition of fallen leaves from the experimental
plants. The VC treated soils showed higher Corg
content throughout which may be due to
stabilization of humified organic compounds in the
vermicast as organo-mineral complex in soil (Li et
al., 2000). Total nitrogen (Ntot) of experimental soil
was higher after the fertilization with VC and IF
(Table 10). In the case of IF treated soils, higher
Ntot was in 15-30 cm layer from the surface than the
first 15 cm layer. The VC treated soil had higher
Ntot in surface soil layer during the first two months
and then the deeper soil layer showed high Ntot
content. The slow release of nutrients by the VC
might have caused higher Ntot retention in surface
soil layer during first two months. Later, increase of
Ntot in deeper soil layers of VC treatments may be
probably due to symbiotic nitrogen fixation by
rhizobia in the roots of the leguminous
experimental plants. These findings are also
supported by higher stimulation of nodulation in
plants grown in experimental soil with VC than
with IF and control.
The ammonium (NH4+) and nitrate (NO3
-)
content of soil were also significantly higher in VC
treatments than the IF treatments and control
(p<0.001; Tables 11-12). Ipomoea based VC
increased the soil NH4+ to the extent of 91% than
the control; and those of lantana and paper waste
based VC showed up to 88 and 74% increase,
respectively. The higher mineral nitrogen content in
VC treated soil was probably the reason of its
higher Corg content which could have provided a
larger source of N for mineralization. Hence, the
VC based treatments might have produced more
residual N in soil than the inorganic fertilizer
(Nethra et al., 1999; Arancon et al., 2006). As the
number of days progressed, the NH4+ content of the
soil reduced to 80% with all the three VC
treatments. In the case of IF treated soils, the NH4+
reduced to the extent of 87%. The NO3- content of
experimental soil also showed similar trend of
results with both VC and IF treatments. In both VC
and IF treatments, the NO3- content reduced up to
46% at the end of the experiment, and the reduction
was much pronounced in ipomoea based VC and its
equivalent IF treatments.
The lantana and ipomoea VC and its
equivalent IF application increased the soil
extractable phosphorus (Pext) content to the extent
of 10% and those of paper waste VC and IF showed
up to 3% (Table 13). As the days progressed, the
Pext content of soil reduced in all the treatments,
which was much pronounced in IF treatments. In all
the VC and IF treatments the higher Pext reduction
was recorded in 15-30 cm soil layer from the
surface than the first 15 cm layer. In the deeper soil
layer, the extensive root of the plants grown in
experimental soil might have uptaken the Pext
maximally resulting in higher Pext depletion in this
layer. The extractable potassium (Kext) content of
soil increased to the extent of 8% after fertilization
with both VC and IF (Table 14). As the days
progressed, the Kext content of soils showed
declining trend with all the treatments. At the end
of experiment, higher reduction in Kext content was
in IF treatments than the corresponding VC
treatments. Among the different VC treatments, the
lantana based VC showed higher reduction of up to
23% in Kext content and in its equivalent IF
treatments, it was up to 25% of reduction. The
reduction of Kext in the ipomoea and paper waste
based VC and its equivalent IF treatments were up
to a maximum of 20%.
Extractable calcium (Caext) content of soil
showed higher reduction to the extent of 45 and
50% with lantana based VC and its equivalent IF
treatments, respectively (Table 15). The paper
waste and ipomoea based treatments showed
maximum reduction of 45%. In all the VC
treatments, the first 15 cm layer of soils showed
higher Caext content than the corresponding IF
144
Table 9. Changes in the organic carbon (g kg−1
) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or equivalent
inorganic fertilizers (IF), at different periods of time (mean ± SD).
Treatments Lantana Ipomoea Paper waste
July Aug Sep July Aug Sep July Aug Sep
Layer
0-15 cm VC 25.0±4.6 27.1±4.9 32.7±6.0 23.5±5.7 27.2±4.3 30.0±5.1 18.0±3.3 21.7±4.0 24.8±4.5
IF 14.6±1.4 19.6±3.1 23.6±2.7 17.3±2.0 18.0±2.3 22.5±2.7 13.6±1.0 15.9±1.6 18.5±1.3
Cont. 16.8±2.0 17.5±1.4 18.7±1.1 16.8±2.0 17.5±1.4 18.7±1.1 16.8±2.0 17.5±1.4 18.7±1.1
15-30 cm VC 20.7±3.2 23.3±3.6 27.9±4.8 18.7±5.9 22.3±5.5 25.1±4.7 14.1±3.3 17.1±2.7 21.0±3.9
IF 13.9±1.8 16.2±2.0 20.4±2.0 13.5±1.4 16.2±1.3 22.0±2.8 13.1±2.2 13.5±2.0 16.5±1.3
Cont. 13.2±1.2 16.2±1.7 20.0±1.3 13.2±1.2 16.2±1.7 20.0±1.3 13.2±1.2 16.2±1.7 20.0±1.3
F value 27.71***
27.87***
25.89***
7.829***
20.33***
17.83***
8.617***
14.33***
15.89***
Amount
5 t ha-1
VC 18.8±2.2 20.8±2.5 24.2±3.1 16.2±2.4 21.6±2.1 23.6±1.6 14.2±4.0 15.7±2.4 18.4±2.1
IF 13.2±1.3 16.5±2.5 20.8±1.9 15.2±2.0 17.1±2.2 25.2±1.7 12.7±1.1 14.1±1.6 17.9±1.1
7.5 t ha-1
VC 23.5±2.5 26.4±2.3 32.8±3.2 18.9±4.0 21.9±5.0 25.5±4.5 15.8±3.3 19.8±2.7 23.0±2.5
IF 14.7±0.8 18.3±3.0 21.3±2.5 16.0±3.6 16.4±1.0 19.6±0.4 14.7±2.2 15.0±2.6 18.1±1.4
10 t ha-1
VC 26.3±4.6 28.4±4.8 33.9±5.3 28.3±3.2 30.8±2.1 33.7±2.8 18.2±3.5 22.7±3.9 27.4±3.3
IF 14.8±2.1 19.0±3.5 23.9±3.2 15.1±2.2 17.7±2.6 22.0±1.6 12.7±0.7 15.1±2.3 16.6±2.0
Control
15.0±2.4 16.8±1.6 19.4±1.3 15.0±2.4 16.8±1.6 19.4±1.3 15.0±2.4 16.8±1.6 19.4±1.3
F value
30.86***
38.29***
43.49***
19.36***
43.42***
53.47***
6.05***
15.57***
35.78***
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
145
Table 10. Changes in the total nitrogen (g kg−1
) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or equivalent
inorganic fertilizers (IF), at different periods of time (mean ± SD).
Treatments Lantana Ipomoea Paper waste
July Aug Sep July Aug Sep July Aug Sep
Layer
0-15 cm VC 7.4±4.0 7.6±4.9 3.8±2.3 7.5±3.7 5.8±3.4 3.3±1.2 4.3±1.7 3.9±1.7 2.7±0.7
IF 7.0±4.9 6.4±4.9 1.7±0.9 6.7±4.4 5.5±4.1 2.1±0.9 5.8±3.1 5.2±3.6 2.7±1.0
Cont. 0.4±0.1 0.7±0.2 0.5±0.2 0.4±0.1 0.7±0.2 0.5±0.2 0.4±0.1 0.7±0.2 0.5±0.2
15-30 cm VC 2.8±1.3 3.3±1.1 4.1±1.4 4.1±2.2 3.6±1.5 4.8±1.7 1.9±0.9 2.2±0.9 1.8±0.4
IF 5.9±3.2 5.4±2.9 2.7±0.9 5.8±2.8 4.0±2.7 2.2±0.9 4.3±2.4 3.6±2.3 2.3±1.2
Cont. 0.4±0.1 1.2±0.4 1.5±0.4 0.4±0.1 1.2±0.4 1.5±0.4 0.4±0.1 1.2±0.4 1.5±0.4
F value 5.954**
5.528**
11.47***
4.173** 4.072**
17.27***
8.311***
5.822**
8.263***
Amount
5 t ha-1
VC 2.8±1.2 3.0±0.7 2.5±0.6 3.6±1.2 2.7±0.7 2.7±1.0 1.9±0.7 2.0±0.7 1.6±0.3
IF 2.1±1.1 3.0±1.3 1.5±0.6 2.5±0.8 2.0±0.9 2.0±0.8 1.8±0.7 1.6±0.8 1.2±0.3
7.5 t ha-1
VC 4.2±2.2 4.7±1.8 3.4±0.9 4.6±2.3 4.1±1.3 3.9±0.7 2.8±1.5 2.9±1.4 2.4±0.6
IF 6.6±0.9 5.0±2.0 2.2±1.0 6.2±0.9 4.8±2.1 1.7±0.5 5.7±1.6 4.7±2.5 2.6±0.7
10 t ha-1
VC 8.4±4.5 8.7±5.7 6.0±1.7 9.2±3.3 7.2±3.3 5.6±1.7 4.7±1.8 4.3±1.7 2.7±0.6
IF 10.7±2.9 9.7±4.3 2.9±1.0 10.1±3.0 7.5±4.2 2.8±0.9 7.6±1.6 6.9±2.9 3.6±0.4
Control
0.4±0.1 0.9±0.4 1.0±0.6 0.4±0.1 0.9±0.4 1.0±0.6 0.4±0.1 0.9±0.4 1.0±0.6
F value
15.45***
9.304***
27.08***
24.59***
10.25***
29.18***
23.57***
11.71***
31.49***
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
146
Table 11. Changes in the ammonium (mg kg−1
) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or equivalent
inorganic fertilizers (IF), at different periods of time (mean ± SD).
Treatments
Lantana Ipomoea Paper waste
July Aug Sep July Aug Sep July Aug Sep
Layer
0-15 cm VC 24.5±11.2 27.2±18.2 7.1±6.1 33.8±19.2 30.3±19.7 6.5±4.5 12.3±5.1 12.1±7.1 3.2±2.6
IF 22.9±10.8 17.4±9.0 4.7±3.1 25.3±12.0 18.1±10.6 3.7±2.9 17.3±7.7 16.3±9.7 4.9±3.9
Cont. 2.2±0.9 0.6±0.2 0.6±0.2 2.2±0.9 0.6±0.2 0.6±0.2 2.2±0.9 0.6±0.2 0.6±0.2
15-30 cm VC 15.3±9.2 13.9±8.6 3.8±3.8 19.5±8.2 16.7±13.4 2.2±1.6 6.8±3.8 7.0±4.7 1.2±1.0
IF 12.7±5.2 8.7±6.1 1.4±0.9 15.0±6.7 10.4±7.0 1.7±1.6 9.3±4.2 8.4±5.5 1.6±1.4
Cont. 0.6±0.1 0.3±0.1 0.2±0.1 0.6±0.1 0.3±0.1 0.2±0.1 0.6±0.1 0.3±0.1 0.2±0.1
F value 8.024***
8.939***
7.134***
7.886***
7.671***
10.07***
10.42***
8.199***
8.217***
Amount
5 t ha-1
VC 10.0±5.3 9.9±6.8 2.1±1.4 19.1±8.2 16.3±9.4 3.1±2.5 6.2±3.7 6.9±4.8 0.9±0.8
IF 13.4±7.6 11.4±8.0 3.0±3.5 15.2±8.0 11.7±6.1 2.1±1.8 10.0±4.5 7.4±5.5 1.5±1.2
7.5 t ha-1
VC 21.3±9.0 21.1±13.1 4.3±3.4 22.5±14.7 20.6±16.3 3.5±3.4 8.8±4.3 8.7±5.6 2.3±1.8
IF 16.4±7.5 11.6±6.7 3.2±2.6 17.8±8.9 11.6±5.8 2.3±2.4 12.5±6.6 13.5±7.8 4.2±4.0
10 t ha-1
VC 28.4±9.9 30.6±18.1 10.0±6.2 38.5±18.6 33.6±22.5 6.4±5.2 13.7±5.1 13.0±7.6 3.4±2.9
IF 23.5±12.0 16.2±11.1 2.9±2.6 27.5±12.4 19.5±13.7 3.8±2.9 17.4±9.0 16.1±10.5 4.1±3.5
Control
1.4±1.0 0.4±0.2 0.4±0.3 1.4±1.0 0.4±0.2 0.4±0.3 1.4±1.0 0.4±0.2 0.4±0.3
F value
7.393***
6.507***
8.131***
5.877***
5.202***
3.792**
5.648***
4.928**
3.959***
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
147
Table 12. Changes in the nitrate (mg kg−1
) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or equivalent
inorganic fertilizers (IF), at different periods of time (mean ± SD).
Treatments Lantana Ipomoea Paper waste
July Aug Sep July Aug Sep July Aug Sep
Layer
0-15 cm VC 121.5±13.4 116.2±19.3 76.5±15.7 127.0±15.4 99.9±18.1 68.4±17.7 96.1±13.0 81.4±8.1 58.9±11.8
IF 116.8±17.2 112.3±30.9 73.5±15.6 114.7±18.0 110.6±31.1 67.3±13.6 98.8±12.5 99.1±16.4 66.8±12.1
Cont. 43.3±4.7 32.3±2.4 23.3±2.6 43.3±4.7 32.3±2.4 23.3±2.6 43.3±4.7 32.3±2.4 23.3±2.6
15-30 cm VC 106.0±12.7 103.0±15.3 70.0±11.9 108.3±7.5 97.7±13.5 63.3±14.9 86.3±7.9 77.4±9.3 57.0±12.1
IF 103.2±16.2 95.2±15.8 66.3±9.5 100.4±17.3 94.4±18.9 62.2±11.8 88.5±7.9 86.8±9.0 62.5±12.6
Cont. 39.7±5.4 28.2±1.9 23.1±3.6 39.7±5.4 28.2±1.9 23.1±3.6 39.7±5.4 28.2±1.9 23.1±3.6
F value 34.78***
27.63***
33.45***
36.50***
22.52***
20.91***
35.51***
56.18***
25.79***
Amount
5 t ha-1
VC 103.5±11.1 98.3±9.1 66.4±8.7 108.0±10.0 84.4±7.7 53.9±11.0 84.6±5.2 71.5±7.7 54.3±10.7
IF 97.9±11.9 86.4±5.7 63.9±8.1 95.5±11.9 85.8±7.8 60.6±9.3 86.9±8.1 85.6±10.7 61.1±11.0
7.5 t ha-1
VC 113.4±7.3 104.0±10.8 72.7±11.6 116.6±9.6 97.7±13.5 66.8±12.6 90.5±9.8 79.6±6.0 55.6±11.8
IF 107.1±15.7 97.9±15.4 65.9±8.5 99.6±13.5 93.3±13.4 62.0±10.0 91.1±10.2 91.3±12.7 63.4±11.5
10 t ha-1
VC 124.4±18.0 126.5±19.7 80.7±17.8 128.4±18.4 114.4±6.8 76.9±16.8 98.6±14.7 87.0±4.6 64.1±11.5
IF 125.0±14.8 127.0±29.6 79.9±15.9 127.6±11.1 128.4±29.7 71.6±16.1 102.9±10.6 102.1±15.5 69.4±14.0
Control
41.5±4.9 30.3±2.9 23.2±3.0 41.5±4.9 30.3±2.9 23.2±3.0 41.5±4.9 30.3±2.9 23.2±3.0
F value
30.11***
34.13***
28.78***
37.02***
37.76***
21.36***
26.77***
46.71***
19.38***
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
148
Table 13. Changes in the exchangeable phosphorus (mg kg−1
) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste,
or equivalent inorganic fertilizers (IF), at different periods of time (mean ± SD).
Treatments Lantana Ipomoea Paper waste
July Aug Sep July Aug Sep July Aug Sep
Layer
0-15 cm VC 4.35±0.31 4.00±0.26 3.64±0.25 4.11±0.22 4.41±0.67 3.60±0.54 4.08±0.18 4.04±0.29 3.75±0.29
IF 4.07±0.43 4.04±0.41 3.58±0.37 4.38±0.48 4.17±0.52 3.54±0.31 4.10±0.08 3.85±0.35 3.57±0.39
Cont. 3.98±0.38 3.81±0.24 3.61±0.14 3.98±0.38 3.81±0.24 3.61±0.14 3.98±0.38 3.81±0.24 3.61±0.14
15-30 cm VC 4.30±0.47 3.89±0.38 3.43±0.29 4.43±0.41 4.21±0.36 3.66±0.31 4.14±0.46 3.72±0.45 3.50±0.33
IF 4.27±0.19 3.77±0.74 3.39±0.43 4.40±0.69 3.86±0.44 3.48±0.27 4.14±0.29 3.97±0.23 3.76±0.26
Cont. 4.11±0.47 3.40±0.45 2.99±0.26 4.11±0.47 3.40±0.45 2.99±0.26 4.11±0.47 3.40±0.45 2.99±0.26
F value 17.72***
5.795**
0.864n.s
16.01***
5.341**
1.124n.s
16.56***
16.45***
1.471n.s
Amount
5 t ha-1
VC 4.34±0.40 3.74±0.36 3.52±0.33 4.29±0.41 4.24±0.59 3.46±0.39 3.92±0.40 3.93±0.50 3.56±0.26
IF 3.89±0.38 3.36±0.55 3.13±0.39 3.90±0.56 3.84±0.57 3.50±0.27 4.03±0.17 3.82±0.43 3.40±0.31
7.5 t ha-1
VC 4.26±0.28 4.05±0.32 3.51±0.29 4.25±0.40 4.40±0.51 3.64±0.36 4.38±0.28 3.79±0.38 3.54±0.44
IF 4.28±0.28 4.22±0.44 3.63±0.28 4.54±0.48 4.04±0.58 3.63±0.28 4.13±0.30 3.94±0.23 3.69±0.32
10 t ha-1
VC 4.38±0.51 4.05±0.19 3.59±0.26 4.29±0.33 4.29±0.56 3.78±0.52 4.04±0.16 3.92±0.36 3.79±0.22
IF 4.34±0.16 4.14±0.41 3.70±0.29 4.74±0.35 4.15±0.33 3.40±0.31 4.21±0.12 3.97±0.17 3.89±0.20
Control
4.04±0.39 3.60±0.40 3.30±0.38 4.04±0.39 3.60±0.40 3.30±0.38 4.04±0.39 3.60±0.40 3.30±0.38
F value
17.49***
6.239***
0.982n.s
24.98***
3.371**
1.374n.s
21.01***
11.75***
0.998n.s
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
149
Table 14. Changes in the exchangeable potassium (g kg−1
) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or
equivalent inorganic fertilizers (IF), at different periods of time (mean ± SD).
Treatments Lantana Ipomoea Paper waste
July Aug Sep July Aug Sep July Aug Sep
Layer
0-15 cm VC 0.42±0.02 0.38±0.02 0.36±0.01 0.41±0.03 0.39±0.02 0.36±0.02 0.41±0.02 0.38±0.02 0.36±0.02
IF 0.40±0.02 0.36±0.02 0.34±0.02 0.41±0.02 0.38±0.02 0.35±0.02 0.40±0.03 0.38±0.01 0.36±0.01
Cont. 0.40±0.01 0.36±0.02 0.34±0.01 0.40±0.01 0.36±0.02 0.34±0.01 0.40±0.01 0.36±0.02 0.34±0.01
15-30 cm VC 0.41±0.02 0.37±0.01 0.35±0.01 0.40±0.02 0.38±0.02 0.35±0.02 0.40±0.01 0.36±0.03 0.34±0.02
IF 0.42±0.02 0.37±0.02 0.33±0.03 0.40±0.03 0.39±0.02 0.36±0.01 0.40±0.02 0.38±0.01 0.35±0.01
Cont. 0.39±0.02 0.35±0.01 0.33±0.01 0.39±0.02 0.35±0.01 0.33±0.01 0.39±0.02 0.35±0.01 0.33±0.01
F value 2.846* 15.73
*** 19.93
*** 2.473
n.s 15.54
*** 18.62
*** 1.799
n.s 18.86
*** 28.68
***
Amount
5 t ha-1
VC 0.40±0.02 0.37±0.01 0.35±0.01 0.40±0.02 0.38±0.03 0.34±0.02 0.39±0.02 0.35±0.03 0.32±0.01
IF 0.39±0.02 0.35±0.01 0.31±0.01 0.39±0.02 0.38±0.01 0.35±0.02 0.39±0.01 0.38±0.01 0.36±0.01
7.5 t ha-1
VC 0.42±0.02 0.38±0.02 0.35±0.02 0.41±0.03 0.40±0.02 0.36±0.02 0.40±0.01 0.39±0.01 0.37±0.01
IF 0.41±0.02 0.36±0.02 0.33±0.01 0.41±0.01 0.38±0.01 0.35±0.02 0.40±0.02 0.38±0.01 0.35±0.01
10 t ha-1
VC 0.43±0.02 0.38±0.01 0.36±0.02 0.41±0.02 0.39±0.02 0.37±0.02 0.41±0.02 0.38±0.02 0.36±0.02
IF 0.42±0.02 0.38±0.01 0.35±0.02 0.42±0.02 0.39±0.03 0.36±0.02 0.41±0.02 0.38±0.01 0.35±0.01
Control
0.39±0.01 0.36±0.02 0.33±0.01 0.39±0.01 0.36±0.02 0.33±0.01 0.39±0.01 0.36±0.02 0.33±0.01
F value
2.142***
11.66***
26.79***
1.020n.s
9.598***
11.91***
1.015n.s
6.576***
17.52***
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
150
Table 15. Changes in the exchangeable calcium (g kg−1
) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or
equivalent inorganic fertilizers (IF), at different periods of time (mean ± SD).
Treatments Lantana Ipomoea Paper waste
July Aug Sep July Aug Sep July Aug Sep
Layer
0-15 cm VC 8.20±0.82 7.11±1.00 5.73±1.02 7.78±1.17 6.66±1.69 5.58±0.91 7.91±0.72 7.15±1.08 6.40±0.95
IF 8.04±0.54 6.64±0.61 5.60±0.89 6.93±1.03 6.62±1.08 5.67±0.87 7.97±0.57 6.55±0.60 5.38±0.52
Cont. 6.32±1.69 5.15±0.45 4.81±1.14 6.32±1.69 5.15±0.45 4.81±1.14 6.32±1.69 5.15±0.45 4.81±1.14
15-30 cm VC 7.79±0.84 5.88±0.82 5.07±0.77 7.05±1.03 5.66±1.09 5.66±0.87 7.18±1.14 5.44±0.74 4.19±0.61
IF 7.61±0.87 6.21±0.97 5.31±0.99 7.72±0.63 6.45±1.42 5.94±1.12 7.92±0.60 5.78±0.76 4.74±0.62
Cont. 5.81±1.58 5.29±0.74 4.55±0.88 5.81±1.58 5.29±0.74 4.55±0.88 5.81±1.58 5.29±0.74 4.55±0.88
F value 2.409n.s
9.511***
24.44***
5.464**
2.488n.s
8.956***
12.49***
17.74***
17.90***
Amount
5 t ha-1
VC 7.41±0.84 5.97±1.05 5.13±0.90 6.81±1.54 5.60±1.36 5.26±0.91 6.96±1.20 5.99±0.96 4.60±1.13
IF 7.37±0.63 5.97±0.97 4.73±0.69 6.62±1.07 5.82±0.81 5.67±0.92 7.81±0.53 5.87±0.87 4.91±0.52
7.5 t ha-1
VC 7.97±0.47 6.69±1.10 5.64±1.19 7.64±1.05 6.18±1.36 5.92±1.03 7.52±0.84 6.53±1.25 5.84±1.27
IF 7.89±0.88 6.35±0.71 5.23±0.39 7.34±0.63 6.93±1.30 5.94±0.94 7.66±0.68 6.19±0.71 5.13±0.78
10 t ha-1
VC 8.60±0.75 6.83±1.06 5.43±0.75 7.77±0.49 6.69±1.67 5.67±0.60 8.16±0.61 6.36±1.60 5.44±1.52
IF 8.21±0.49 6.95±0.47 6.40±0.76 8.03±0.42 6.86±1.34 5.80±1.18 8.37±0.13 6.44±0.73 5.14±0.66
Control
6.06±1.49 5.22±0.57 4.68±0.97 6.06±1.49 5.22±0.57 4.68±0.97 6.06±1.49 5.22±0.57 4.68±0.97
F value
8.544***
11.47***
13.7`***
3.884**
0.908n.s
4.822***
31.33***
19.17***
14.93***
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
151
Table 16. Changes in the micronutrient content (mg kg−1
) of potting soil amended with vermicast (VC) from lantana or equivalent inorganic
fertilizers (IF), at different periods of time (mean ± SD).
Treatments Mg Cu Fe Mn Zn B
Initial Final Initial Final Initial Final Initial Final Initial Final Initial Final
Layer
0-15 cm VC 99.8 63.1 6.55 2.76 59.3 52.8 65.0 36.8 58.7 26.7 28.3 12.4
IF 98.4 49.0 6.10 1.48 58.3 46.7 60.3 33.0 57.4 24.9 28.1 10.3
Cont. 88.0 43.4 5.09 1.24 55.4 49.4 46.2 37.4 53.5 25.5 26.0 13.8
15-30 cm VC 91.9 48.6 5.25 2.10 56.9 44.2 52.2 29.4 56.0 24.3 25.8 11.2
IF 88.2 56.3 5.52 1.80 55.4 40.5 54.3 30.9 56.3 24.1 26.2 10.5
Cont. 86.3 40.9 5.04 3.45 58.3 54.6 46.0 28.2 54.8 22.2 27.2 14.9
F value 5.833**
7.054**
18.47***
34.16***
2.243n.s
2.497n.s
13.78***
7.194**
8.757**
3.569* 2.668
n.s 1.447
n.s
Amount
5 t ha-1
VC 91.3 50.5 5.67 2.30 56.4 40.5 54.2 30.7 56.6 24.9 26.7 12.4
IF 88.9 51.5 5.58 1.24 55.2 44.7 53.8 29.8 55.7 23.9 26.2 13.0
7.5 t ha-1
VC 93.8 53.0 5.79 2.50 57.5 46.7 58.1 36.5 57.4 25.7 26.7 11.1
IF 93.6 53.5 5.83 1.77 56.7 40.6 57.3 31.3 57.2 24.7 27.8 9.5
10 t ha-1
VC 102.4 64.1 6.23 2.50 60.2 58.3 63.4 32.2 58.1 25.9 27.7 11.9
IF 97.4 53.0 6.02 1.91 58.5 45.4 60.8 34.8 57.8 24.9 27.4 8.7
Control
87.2 42.2 5.06 2.35 56.9 52.0 46.1 32.8 54.1 23.9 26.6 14.4
F value 3.413* 2.788
n.s 1.582
n.s 0.640
n.s 1.914
n.s 3.818
* 4.430
* 0.430
n.s 3.627
* 0.672
n.s 0.364
n.s 3.404
n.s
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
152
Table 17. Changes in the micronutrient content (mg kg−1
) of potting soil amended with vermicast (VC) from ipomoea or equivalent inorganic
fertilizers (IF), at different periods of time (mean ± SD).
Treatments Mg Cu Fe Mn Zn B
Initial Final Initial Final Initial Final Initial Final Initial Final Initial Final
Layer
0-15 cm VC 103.9 50.7 5.42 2.67 72.1 41.4 55.8 25.6 56.9 25.6 31.0 15.2
IF 98.0 44.4 5.35 1.75 69.4 49.7 53.1 31.0 55.6 27.4 31.3 14.6
Cont. 88.0 43.4 5.09 1.24 55.4 49.4 46.2 37.4 53.5 25.5 26.0 13.8
15-30 cm VC 90.3 40.6 5.08 2.78 56.7 32.0 50.1 21.5 55.0 23.6 26.4 11.9
IF 88.6 40.7 5.07 2.30 57.0 49.0 48.8 26.3 54.5 24.1 27.0 14.4
Cont. 86.3 40.9 5.04 3.45 58.3 54.6 46.0 28.2 54.8 22.2 27.2 14.9
F value 3.600* 5.958
** 2.129
n.s 18.86
*** 20.27
*** 6.841
** 6.324
** 33.98
*** 3.544
* 3.071
n.s 4.668
* 1.110
n.s
Amount
5 t ha-1
VC 86.3 46.4 5.17 2.32 61.7 32.0 49.8 25.1 54.5 22.9 27.2 14.2
IF 89.0 40.0 5.41 2.12 60.9 52.8 48.0 27.8 53.8 26.8 27.1 15.4
7.5 t ha-1
VC 99.0 44.1 5.24 2.98 63.7 41.2 52.8 21.4 56.6 24.8 28.3 14.5
IF 92.4 44.2 5.10 2.13 64.2 52.0 50.3 29.9 55.1 25.4 29.2 14.6
10 t ha-1
VC 106.0 46.6 5.34 2.88 67.7 36.8 56.3 24.2 56.6 26.1 30.6 11.9
IF 98.7 43.4 5.11 1.81 64.6 43.3 54.5 28.1 56.1 25.0 31.1 13.5
Control
87.2 42.2 5.06 2.35 56.9 52.0 46.1 32.8 54.1 23.9 26.6 14.4
F value 3.552* 0.514
n.s 0.722
n.s 0.503
n.s 1.065
n.s 5.443
** 5.502
** 1.846
n.s 2.186
n.s 1.003
n.s 1.186
n.s 0.605
n.s
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
153
Table 18. Changes in the micronutrient content (mg kg−1
) of potting soil amended with vermicast (VC) from paper waste or equivalent inorganic
fertilizers (IF), at different periods of time (mean ± SD).
Treatments
Mg Cu Fe Mn Zn B
Initial Final Initial Final Initial Final Initial Final Initial Final Initial Final
Layer
0-15 cm VC 105.7 57.4 5.47 2.52 65.8 35.8 52.5 36.2 56.5 22.2 26.4 6.2
IF 105.2 58.3 5.18 2.67 63.6 33.2 55.4 31.9 55.8 23.7 26.5 7.4
Cont. 88.0 43.4 5.09 1.24 55.4 49.4 46.2 37.4 53.5 25.5 26.0 13.8
15-30 cm VC 90.2 54.1 5.09 2.04 59.1 31.0 47.8 28.6 54.9 21.5 25.3 6.2
IF 87.6 43.4 5.15 1.37 58.1 28.0 49.5 29.8 55.0 21.7 26.3 11.7
Cont. 86.3 40.9 5.04 3.45 58.3 54.6 46.0 28.2 54.8 22.2 27.2 14.9
F value 4.861* 2.601
n.s 3.406
* 35.22
*** 8.143
** 68.54
*** 10.49
*** 7.154
** 3.785
* 2.894
n.s 0.875
n.s 3.664
*
Amount
5 t ha-1
VC 93.2 51.3 5.17 2.03 60.0 33.2 48.3 29.6 54.4 21.7 25.7 7.9
IF 90.0 45.5 5.07 1.77 57.9 29.6 50.1 28.2 54.8 22.1 26.7 10.3
7.5 t ha-1
VC 93.4 56.5 5.42 2.24 62.1 32.9 50.7 31.4 56.3 22.2 25.2 5.9
IF 94.8 51.9 5.11 2.04 61.5 28.8 53.3 30.7 55.0 22.7 26.0 11.1
10 t ha-1
VC 107.1 59.4 5.25 2.56 65.2 34.1 51.6 36.1 56.4 21.5 26.7 4.8
IF 104.3 55.3 5.31 2.24 63.1 33.4 54.0 33.7 56.2 23.4 26.5 7.2
Control
87.2 42.2 5.06 2.35 56.9 52.0 46.1 32.8 54.1 23.9 26.6 14.4
F value 1.746n.s
1.275n.s
1.352n.s
0.151n.s
3.043n.s
17.23***
3.248* 0.540
n.s 2.065
n.s 0.797
n.s 0.920
n.s 2.721
n.s
*p<0.05,
**p<0.01,
***p<0.001,
n.s. not significant
154
treatments; whereas, in the 15–30 cm layer a higher
Caext content was recorded with IF treatments. The
concentration of trace nutrients such extractable
form of magnesium (Mgext), copper (Cuext), iron
(Feext), manganese (Mnext), zinc (Znext) and boron
(Bext) significantly increased in soil with both VC
and IF fertilization (Tables 16-18). The Mgext
content in soil increased to the extent of 19% with
paper waste based VC and its equivalent IF
treatments. Increase in Mgext with ipomoea and
lantana based treatments were up to 18 and 15%,
respectively. At the end of the experiment, the
Mgext with ipomoea based treatments showed
higher reduction of 56% followed by 50 and 46%
with paper waste and lantana based treatments.
Fertilization with VC and IF, increased the Cuext
content to the extent of 70%. Finally, a maximum
reduction of about 19% in Cuext content was
recorded in lantana based treatments; whereas, the
paper waste and ipomoea based treatments showed
only 7% maximally.
Lantana, paper waste and ipomoea based VC
and its equivalent IF treatments increased the Feext
content of soil to the extent of 23, 16 and 7%,
respectively. In the case of Znext up to 7, 4.4 and
4.1% increase was recorded with lantana, ipomoea
and paper waste based treatments. Except the
ipomoea based treatments, in all others, the Feext
and Znext depletion was lower in the VC compared
to corresponding IF treated soils. Similarly, the
ipomoea and paper waste based VC treatments
showed lesser reduction in Mnext compared to its
equivalent IF treatments; whereas, the lantana
based VC showed higher reduction. By the end of
the experiment, the reduction in Mnext was to the
extent of 60, 59 and 43% with ipomoea, lantana and
paper waste based treatments, respectively. The
increase of about 14.5% in Bext content was in
lantana based treatments, and those of ipomoea and
paper waste showed 3 and 0.4%, respectively. The
higher Bext content in lantana based treatments
could be the reason for better growth of
experimental plants with maximum nodulation.
Increase in availability of Bext was reported to
enhance N fixation in the nodules of soybeans
(Yamagishi and Yamamoto, 1994). Reduction in
Bext content after plant harvest was 82, 68 and 61%
with paper waste, lantana and ipomoea based
treatments, respectively. The trace nutrients, such
as Cuext, Feext, Znext and Bext depleted maximally in
IF based treatments, in both surface and deeper soil
layer; whereas Mgext and Mnext content of surface
soil layers reduced maximally with IF treatments
and in the deeper soil layer VC showed higher
reduction. In all the cases, the increasing dose of
nutrient application exaggerated its changes as
stated above, and there was no differential trend of
results with different dose of VC and IF
application.
The nutrient status of soil is influenced by
several factors. Amongst the nutrient up take by
plants, nutrient leaching from soil, microbial
immobilization and mineralization are considered
to influence the fate of nutrients in the soil largely
(Carlile and Wilson, 1993). In these experiments,
lower concentration of many nutrients in IF treated
soils may be due to high leaching of mineral
nutrients from the soil. Organic matter inputs
through VC, in addition to supplying nutrients,
improve soil aggregation, and stimulate microbial
diversity and activity (Shiralipour et al., 1992;
Carpenter-Boggs et al., 2000). The changes in the
physical and microbial properties of soil influence
many chemical and biological reactions (Sharma
and Bhushan, 2001). In the present study, increase
in microbial and enzymatic activity with
vermicompost application (Masciandaro et al.,
1997; Arancon et al., 2006) might have mineralized
the organic bound nutrients, which attributed to
increase in the mineral nutrient content in soil
together with improved growth and productivity of
experimental plants.
4. Conclusions
The paper describes the impact of
vermicompost derived from different parent
materials on the physical and chemical qualities of
155
soil which are directly related to the plant growth.
The impact of the vermicompost on soil health in
terms of its physical and chemical qualities were
compared with that of an inorganic fertilizers which
had all the main macro- and micro-nutrients in
concentrations equivalent to the ones present in the
vermicompost. Several sets of experiments were
carried out in which the vermicast generated from
different organic wastes such as paper waste, leaves
of ipomoea (Ipomoea carnea), and of lantana
(Lantana camara) or inorganic fertilizers were
applied in potting soil housing cluster bean
(Cyamopsis tetragonoloba). Samples from all these
treatments were collected on weekly basis during
different stages of plant growth. The results reveal
that vermicast application created a suitable
physical environment by reduction in bulk density
and improving the water holding capacity and
porosity of soil used. In addition, throughout the
experiment, the nutrient content of soil was
significantly higher compared to inorganic
fertilizers treated one. Although, the inorganic
fertilizers application initially increased the nutrient
content in soil, as the days progressed, substantial
quantity of applied nutrient became unavailable to
the plants, probably it lost due to high leaching of
mineral nutrients from the soil. Apparently, no
significant impact has been observed on physical
properties of soil with inorganic fertilizers
application. Consequently, the vermicompost
amendment may be considered a good strategy for
improving plant growth which reduces the
deterioration of agricultural lands due to rampant
use of inorganic fertilizers.
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EFFECT OF STORAGE ON SOME PHYSICAL AND
CHEMICAL CHARACTERISTICS OF VERMICAST: A
PRELIMINARY STUDY
Chapter
10
159
A paper based on this chapter has been accepted for publication in
Journal of Applied Horticulture
CChhaapptteerr 1100
Effect of storage on some physical and chemical
characteristics of vermicast: A preliminary study
Abstract
It is widely acknowledged that vermicast has beneficial effect on plant growth but little is known on how the
manner and duration of storage affect the vermicast quality. Investigating the impact of storage on the
characteristics of castings is of great importance with respect to understanding its optimum age for best
utilization. In an attempt to cover this knowledge-gap we have carried out studies on the morphology, water-
retention capacity, and availability of vermicast nutrients as function of vermicast ageing when the latter is
stored. The study reveals that during storage of vermicast, the physical and chemical properties of castings
get altered. The prolonged storage period reduced the nutrient concentration and in turn the beneficial
properties of vermicast required for plant growth.
1. Introduction
Vermicomposting is the term given to the
process of conversion of biodegradable matter by
earthworms into vermicast (Abbasi and Ramasamy,
2001). In the process, a major fraction of the
nutrients contained in the organic matter is
converted to more bioavailable forms. Application
of vermicompost improves the soil structure by
increasing porosity and reducing the bulk density. It
improvises soil aeration, water-holding capacity,
buffer capacity, and cation exchange capacity of
soil (Nada et al., 2011). In addition, the
vermicompost is also reported to contain
biologically active substances such as plant growth
regulators and have been shown to increase growth
of many plants (Tomati et al., 1983, 1988, 1990;
Tomati and Galli, 1995; Abbasi and Ramasamy,
1999; Atiyeh et al., 2002; Arancon et al., 2003,
2004; Gajalakshmi and Abbasi, 2004; Acevedo and
Pire, 2004; Edwards, 2004; Sinha, 2009).
Although a considerable number of studies
have been carried out on vermicomposting of
various organic materials with different earthworm
species and their impact on the soil and plant
growth (Logsdon, 1994; Sundaravadivel and Ismail,
1995; Gajalakshmi et al., 2001a,b, 2002; Singh and
Sharma, 2002; Gajalakshmi and Abbasi, 2003,
2004; Suthar, 2006, 2007; Padmavathiamma et al.,
2008), there is still a lack of knowledge on the
change in the properties of vermicast during the
course of storage. The castings of anecic and
endogeic earthworms have been extensively studied
in relation to the changes in the physico-chemical
properties during ageing process (Hindell et al.,
1997; Decaëns et al., 1999; Decaëns, 2000; Aira et
al., 2005; Mariani et al., 2007). There are also few
studies on the enzymes and microbial aspects of
castings generated from the epigeic earthworms
(Parthasarathi and Ranganathan, 1998, 1999;
Scullion et al., 2003). There is no study on changes
in the characteristics of vermicast during the ageing
as a function of manner of storage. Investigating
this storage effect on castings is of great importance
with respect to understanding the changes in
nutrient status and physical properties of vermicast.
The present work has been taken up to investigate
160
the changes in the properties of the vermicast when
stored so that the optimum age for best utilization
could be understood. The present study was
conducted with vermicast generated from neem
leaves.
2. Materials and methods
Neem leaves were collected from the
Pondicherry University campus and its vicinity.
The collected leaves were washed with water to
remove adhering material and soaked for 48 hours
in order to remove phenolic compounds and to
make substrate softer and palatable to earthworms
(Agarwal et al., 1987; Nath et al., 1987; Agarwal et
al., 1991). Rectangular wooden boxes (depth 30
cm, width 35 cm, length 39 cm) were used as
vermireactors. The reactors were filled from bottom
up with successive layers of coarse sand and soil to
a thickness of 3 and 5 cm, respectively. Neem
leaves was added as feed with an epigeic
earthworm species Eudrilus eugeniae. After 2
weeks, the vermicast was harvested. An aliquot of
fresh castings was analyzed immediately whilst the
rest were stored for <7, 14, 21, 60, 90 and >120
days. The casts were stored in the polyethylene
bags of 20 micron thick and 25 x 18 cm size.
Plastic bags filled with 500 g of vermicast were
stored at room temperature in order to imitate the
general way of storage of vermicomposting in
commercial sectors.
Moisture content of castings was determined
by weight loss at 105°C. To estimate bulk density
(pBulk), sample volume was measured with a
graduated cylinder and its dry weight determined
by oven drying. The particle density (pParticle) was
determined by volumetric flask method (Bashour
and Sayegh, 2007). The quotient value of weight of
the sample and its volume which was measured
through volume of water displaced by known
amount of soil sample in the volumetric flask is
reported as particle density. To measure the water
holding capacity (WHC), the samples were filled in
cylinders with a perforated base and immersed in
water and drained. The quantity of water taken up
by samples is determined by drying to constant
mass at 105°C (Margesin and Schinner, 2005). The
total and water filled porosity were calculated from
the particle and bulk density values of the
respective samples, using the following equation
(Carter and Gregorich, 2008).
….…….…….… (1)
………. (2)
.(3)
were, Db is the bulk density, Dp is the particle
density, θw is gravimetric water content, and Dw is
the density of water at corresponding temperature.
Electrical conductivity (EC) and pH were
measured with suspensions of samples in water
(1:2, w/v) (Bashour and Sayegh, 2007) by using
EI™ 611E EC meter and Digison™ digital pH
meter 7007, respectively. Thin sections of casts
were made after impregnating the samples in
araldite using Bueller PetroThin™ thin sectioning
system (FitzPatrick, 1993). The internal and
external structures of thin sectioned castings were
observed under the binocular microscope.
Total organic carbon (Corg) was determined
following modified dichromate redox method
(Heanes, 1984). External heating was applied
during the oxidation process in order to quicken
and complete oxidation of organic carbon in the
sample. Total nitrogen (Ntot) was determined by
modified Kjeldahl method (Kandeler, 1993) using
Kel Plus™ semi-automated digester and distillation
units. In order to include nitrate, nitrite, nitro and
nitroso groups in the assay, a mixture of salicylic
acid and sulphuric acid was used for digestion. All
the elemental present in the vermicast were
analyzed by Bruker™ S4-Pioneer model
wavelength dispersive X-ray fluorescence
spectrophotometer (WD-XRF). The samples were
ground to particle size well below 100 µm using
ball mill in order to minimize the grain size
interference on XRF-measurement. The
161
concentration of major elements found in the
vermicast is reported in this chapter. The data were
analyzed statistically with software SPSS 16
package and subjected to one-way ANOVA.
Comparisons between means were tested with LSD
test.
3. Results and discussion
Fresh castings of E.eugeniae produced from
neem leaf litter were fine, long, slender and pellet-
like in structure. The size was in the range of 0.1 to
2.0 mm length and 0.1 to 1.2 mm breadth. During
storage, the casts had undergone significant
changes in their physical and chemical properties
(p< 0.001) (Tables 1-2). The castings stored for
prolonged period (>120 days), had its structure
disintegrated and formed clod like aggregates. The
initial moisture content of 77.4 ± 3.5% was reduced
to 66.7±2.5% in 60 days stored vermicast.
Afterwards, the moisture content drastically
decreased to about 20% in the 90 and >120 days of
storage. Decrease in the moisture content of
vermicast during the storage may exert a strong
influence on the microorganisms (Nannipieri et al.,
2003) and their enzyme activity in the vermicast
(Parthasarathi and Ranganathan, 1998). The
microbiota and their enzymes in turn reflect on the
mineralization of nutrient (Birch, 1964). The results
indicate that the drastic loss in moisture content
(>75%) after 90 days leads to reduction in
microbial mediated activity in the vermicast.
In the first 3 weeks of storage, the particle
density (pParticle) was stable, after that it increased
and became twofold high at > 120 days stored
vermicast compared to the fresh castings (p<0.001)
(Table 1). Ruhlmann et al. (2006) has reported that
the pParticle of castings varied considerably due to
the degree of decomposition of organic matter
present in it. In the present study the pParticle was
ranging from 1.359 to 2 g cm-3
, which is very lower
than the soil pParticle range of 2.6–2.8 g cm-3
in
relation to the plant growth. Low pParticle of the
vermicast can be explained by their high Corg
content. It has been reported that increase of Corg in
the soil, decrease the pParticle at the rate of 0.04-0.06
g cm-3
per percentage of Corg (Ruhlmann et al.,
2006).
As the age of the vermicast progressed, there
was a significant increase in bulk density (pBulk)
(p<0.001; Table 1). During the first week of
storage, the pBulk was 0.307 g cm-3
, and then it
increased, till the end of the experiment
(0.603±0.014 g cm-3
). There was twofold increase
in pBulk in four months of storage. However, it is
lower than the soil pBulk range of 0.7–1.8 g cm-3
in
relation to the plant growth (Lal and Shukla, 2004).
Low pBulk is desirable for plant growth, as it makes
easier for plant root penetration and it posses high
water infiltration rates. On the other hand, high pBulk
would impede root penetration and reduce the air
and water movement (Edwards, 2004).
Porosity of castings decreased distinctly
(p<0.001) throughout the >120 days of storage and
its range was between 69.9 to 77.4% (Table 1).
Porosity directly influences water infiltration,
hydraulic conductivity and water storage capacity
in soils (Blanchart et al., 2004). Moreover, it
greatly influences the structure, function and
interaction of microbial and microfaunal
communities (Hattori, 1994). The percentage of
water-filled pore space (WFPS) was high, but not
significantly different in the first 21 days of storage,
ranging between 30.5 and 33.4%. High WFPS may
decline the microbial activity, presumably as a
result of additional water presenting a barrier for
diffusion of oxygen and the waste products away
from microorganisms (Linn and Doran, 1984a,b;
Doran, 1990). The castings stored for 90 and >120
days showed 14.9 and 18.2% of WFPS,
respectively; this lower WFPS is also not suitable
for the microbial activity. Usually excessive
dryness is more prejudicial to microorganisms than
an excess of water-filled pores (Tate, 1985;
Paradelo and Barral, 2009).
WHC was the maximum in castings stored
162
Table 1. Physical characteristics (mean± SD) of castings stored for different periods and the calculated F-
values using one-way ANOVA
*p<0.05,
**p<0.01,
***p<0.001,
n.s - not significant.
for 2 weeks and it was 651.0 ± 37.4 and 798.0 ±
9.0% in the first and second week, respectively
(Table 1). High WHC of the castings during the
initial period of storage may be due to the
abundance of micropores present in the castings
(Chaudhuri et al., 2009). At the end of the
experiment nearly 6 fold decrease in WHC was
recorded compared to fresh castings (p<0.001).
While estimating the WHC of the vermicast stored
for 90 and > 120 days, it was observed that the
vermicast did not absorb water for many hours,
indicating high degree of water repellency. This
may be due to conformational rearrangements of
the organic matter (Mashum and Farmer, 1985;
Valat et al., 1991; Roy et al., 2000), and excess
dryness (King, 1981; Ritsema et al., 1998; De
Jonge et al., 1999; Bachmann and van der Ploeg,
2002; Quyum et al., 2002).
Electrical conductivity indicates the
concentration of total soluble salts in solution, thus
reflecting the degree of soil salinity and it affects
plants at all stages of development. The sensitivity
may vary from one growth stage to another for
some crops (Maas and Hoffman, 1977). In the
neem castings of our study, maximum EC of
4.893±0.210 mmhos cm-1
was recorded in the fresh
castings and it significantly declined during the
storage (p<0.001) (Table 1). There was a maximum
of about 68% reduction in EC during first 21 days
of storage and thereafter there was a slow and
steady decline till the end of the experiment.
Table 2. Total nitrogen and organic carbon (mean± SD) of castings stored for different periods and the
calculated F-values using one-way ANOVA
Parameter Days of storage
F value < 7 14 21 60 90 >120
Total Kjeldahl
nitrogen mg g-1
21.77 ± 1.33 14.95 ± 0.35 14.33 ± 1.46 12.55 ± 2.19 13.94 ± 2.49 11.20 ± 0.83 239.5
***
Total organic carbon
mg g-1
108.0 ± 2.1 115.2 ± 8.7 130.8 ± 2.5 144.6 ± 5.4 151.2 ± 1.7 90.0 ± 1.9 46.5
***
*p<0.05,
**p<0.01,
***p<0.001,
n.s - not significant.
Days of
storage
Water Content
%
Bulk Density
g cm-3
Particle Density
g cm-3
Pore Space
%
Water-Filled
Pore Space
%
Water Holding
Capacity
%
EC
mmhos cm-1
<7 77.44 ± 3.46 0.307 ± 0.006 1.359 ± 0.034 77.41 ± 0.94 30.49 ± 1.77 651.0 ± 37.4 4.893 ± 0.210
14 76.67 ± 0.84 0.317 ± 0.009 1.349 ± 0.33 76.47 ± 0.70 31.63 ± 1.14 798.0 ± 9.0 3.673 ± 0.361
21 75.70 ± 3.86 0.334 ± 0.005 1.350 ± 0.012 75.26 ± 0.48 33.41 ± 2.06 592.1 ± 30.8 1.569 ± 0.023
60 66.73 ± 2.45 0.383 ± 0.014 1.435 ± 0.037 73.31 ± 0.86 34.65 ± 1.57 518.0 ± 20.6 1.526 ± 0.050
90 20.63 ± 3.49 0.524 ± 0.015 1.849 ± 0.028 71.68 ± 0.54 14.87 ± 2.94 132.7 ± 5.0 1.489 ± 0.029
>120 21.36 ± 2.82 0.603 ± 0.014 2.000 ± 0.037 69.86 ± 0.70 18.24 ± 2.94 117.8 ± 9.0 1.439 ± 0.040
F value 772.1***
1034.7***
774.6***
145.9***
135.5***
1441.1***
649.4***
163
Table 3. Major elements present in the casting stored for different periods (expressed in %).
Elements Days of storage
< 7 14 21 60 90 >120
Ca 60.18 60.14 60.09 48.91 17.444 15.56
Al 2.280 1.260 1.190 1.120 1.042 1.045
Cl 3.890 3.850 3.790 3.150 0.898 0.774
Fe 3.773 2.505 2.382 1.878 1.643 1.108
K 12.92 14.77 14.85 10.69 2.580 2.864
Mg 2.578 3.716 3.848 2.443 0.633 1.111
Mn 0.258 0.193 0.179 0.199 0.070 0.245
Na 0.429 0.428 0.428 0.397 0.394 0.436
P 2.090 1.460 1.430 1.430 1.425 1.230
S 5.360 4.169 4.146 4.058 1.060 1.480
Si 15.63 15.54 15.38 15.01 13.38 12.82
Ti 0.888 0.446 0.368 0.133 0.105 0.070
Zn 0.377 0.093 0.090 0.090 Nil Nil
The total nitrogen content in fresh castings
was 21.77±1.33 mg g-1
(Table 2). In the 14 days
stored castings, 31% reduction in Ntot was observed.
Afterwards the concentration decreased slightly
during the entire storage process. The gaseous loss
of nitrogen from the casts may be the reason for the
maximum Ntot loss during 14 days of storage. A
higher loss of N during the initial weeks was likely
due to the intense ammonia volatilization. In
general, NH3+ volatilization is strongly dependent
on the NH3+ and NH4
+ concentration (Pagans et al.,
2006). It has been reported that up to 30% nitrogen
loss occurs in fresh castings by denitrification
(Kharin and Kurakov, 2009). The organic carbon
content of fresh castings was 108.0±2.1 mg g-1
.
During the storage process, the Corg content in the
castings significantly increased up to 29% in the
course of 90 days. After 120 days, there was a
drastic decrease and its concentration was near to
those of fresh castings (Table 2).
Decaëns et al. (1999) observed same pattern
of Corg increase but without any decline over
prolonged ageing of vermicast of large species of
anecic earthworm, Martiodrilus carimaguensis.
They have also summarized a combination of
several factors for Corg increase during the ageing
process. Among those factors, the possible reasons
that would be applicable in the present studies are
fixation of atmospheric CO2 from autotrophic
microorganisms, such as algae or nitrification
microorganisms (Vinceslas- Akpa and Loquet,
1997) and accumulation of organic matter by cast-
dwelling macroinvertebrates. The significant
decrease of Corg in vermicast stored for more than
120 days can be attributed to excessive dryness of
castings, which is not suitable and beneficial to
microbiota. The fresh castings are noted to be
enriched in Ca, followed by K (Table 3). The
concentration of these elements was almost stable
until 21 days of storage. Afterwards, there was a
decline throughout the study. Lal and de
Vleeschauwer, (1981) and Schrader and Zhang
(1997) ascribed the high Ca content of casts to the
presence of calcite spheroids originating from
worms’ calciferous glands which regulates the CO2
in their tissues (Briones et al., 2008).
In the fresh castings, 12.9% of K was
recorded and there was 77% of loss at the end of 4
months storage. In the case of P, the initial
concentration of 2.1% was reduced to 1.2%, at the
end of 4 months. A maximum P loss of 73% was
observed in the first two weeks of storage. After
that, there was slow and steady decline of P content
till the end of the experiment. Except Mg, the
164
concentration of other elements such as Al, Fe, Na,
Cl, S, Si, Ti and Zn decreased as the storage period
increased (Table 3). Changes in Mg concentration
did not show any trend as the age of the castings
progressed. The overall results show that 60 days of
storage did not show much variation; after that
there was a significant decrease in the concentration
of these elements. The reduction of K, P and other
metals content may be due to nutrient assimilation
by the bacterial and fungal grazing macro-
invertebrates in the castings. Loss of these trace
nutrients may slide down the positive impact of
vermicompost on the plant growth.
4. Conclusions
The present study reveals that during storage
of vermicast, the physical and chemical properties
of castings get altered. The prolonged storage
period reduced the nutrient concentration and in
turn the beneficial properties required for plant
growth. According to the results of this study, most
of the characteristics of the castings are retained
during the first 60 days of storage. Further as
storage was continued, the nutrient status depleted.
The changes in physical properties are
disintegration of the structure of the vermicast,
increase in bulk density, water repellency, decrease
in water holding capacity and water content. All
these factors lead to adverse impact on plants when
applied as manure. Therefore, utilization of
vermicast before nutrient loss is recommended or
castings need to be stored by appropriate methods
which should prevent the loss of nutrient
concentration and maintain the physical
characteristics of vermicast. At present, there are no
prescribed guidelines for storage of castings; hence
comprehensive method of storage needs to be
explored extensively.
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168
EFFECT OF STORAGE ON THE PROPERTIES OF
VERMICOMPOST GENERATED FROM PAPER WASTE –
WITH FOCUS ON PRE-DRYING AND EXTENT OF
SEALING
Chapter
11
169
A paper based on this chapter has been published in
International Journal of Energy and Environmental Engineering, DOI: 10.1007/s40095-014-0135-z
CChhaapptteerr 1111
Effect of storage on the properties of vermicompost generated
from paper waste – with focus on pre-drying and extent of
sealing
Abstract
In chapter 10, a preliminary study on the impact of storage of vermicast of different age, without any
pretreatment was reported. The study revealed that the physical and chemical properties of vermicast get
altered during storage thereby reducing the fertilizer value of vermicast. In the present study the effect of pre-
drying and type of packing is reported. Vermicast generated from paper waste was packed in airtight and
partially sealed bags with and without pre-drying for 24 hours. Changes in several physical, chemical, and
biological properties of the castings were monitored for three months with weekly assessments. The results
reveal that the beneficial properties of vermicast were the highest when it was fresh. There was deterioration
on storage, which can be minimized if the castings are contained in airtight bags after pre-drying the casts.
1. Introduction
The vermicast that is deposited by the
earthworms on the soil is known to fertilize the soil
as well as influence its physical and chemical
properties in a way that is beneficial to plant growth
in particular and soil environment in general. Due
to this realization several studies have been
conducted on the fate of vermicast, especially how
the biological, chemical, and physical attributes of
the vermicast change with time (Hindell et al.,
1997a; Decaëns et al., 1999; Parthasarathi and
Ranganathan, 1999, 2000; Decaëns, 2000; Tiunov
and Scheu, 2000; Scullion et al., 2003; Aira et al.,
2005, 2010; Mariani et al., 2007; Kawaguchi et al.,
2011). These studies have been on either vermicast
generated from non-specific substrates in nature or
from blends of soil and phytomass. The focus of the
studies has been primarily on the stability of
vermicast generated by anecic and endogeic
(geophagous and geophytophagous) earthworm
species as such casts are rich in soil and influence
the stability of biogenic structures. Very few
studies have been done on epigeic or phytophagous
(‘humus feeder’) species. Moreover, when
vermicast is deposited in nature, its fate is strongly
influenced by (a) soil dwelling invertebrates –
which colonize the vermicast and feed upon the
organic matter it contains (Decaens et al., 1999;
Decaëns, 2000); (b) vegetation – which takes up the
nutrients from the castings (Jiménez and Decaëns,
2004); (c) soil microbes including autotrophic
algae, nitrification bacteria or fungi – which are
involved in fixation of atmospheric CO2 (Jiménez
and Decaëns, 2004); (d) immobilization or
mineralization of nutrients in the vermicast (Kharin
and Kurakov, 2009), and (e) environmental factors
such as rain, flooding, or drought.
A few controlled studies have been reported
on the change in the properties of vermicast upon
ageing (Shipitalo and Protz, 1989; Marinissen and
Dexter, 1990; Marinissen et al., 1996; Hindell et
al., 1997a,b; McInerney and Bolger, 2000; Tiunov
and Scheu, 2000). These studies have primarily
aimed to simulate the conditions which the biogenic
170
structures experience in nature. For this the casts
were generated from either soil or blends of soil
and phytomass and stored in soil/sand columns. It
was seen that the soil particles present in the casts
get chemically bound with organic matter, perhaps
through chelation, which increases the stability of
the casts. It also protects the organic matter content
of the casts from decomposition (Shipitalo and
Protz, 1989; McInerney et al., 2001), as the organic
matter that is attached to the minerals with strong
chemical bounds is less accessible to
microorganisms (Kaiser et al., 2007). In addition,
extracellular enzymes are protected from
degradation and proteolysis by the clay minerals
contributed by the soil (Nannipieri and Smalla,
2006).
In contrast to the focus of the prior art
summarized as above, the conditions associated
with the storage of vermicast when it is produced
by anthropogenically controlled vermicomposting
and for the specific purpose of use as a fertilizer are
very different. The concern here is to ensure that
the vermicast retains as many and as much of the
plant-friendly attributes as does fresh vermicast and
the physical integrity of the cast is not of much
significance. The only pre-existing studies on the
effect of storage on vermicompost (Parthasarathi
and Ranganathan, 1999, 2000), have been based on
the use of two-month old press-mud as feed for
earthworms, and assessment of the changes in
major nutrients (N,P and K), microbial activity and
enzyme activity of the vermicast that was
generated. In these studies, the environmental
conditions under which the casts have been exposed
during the ageing – either in vermireactors or in a
controlled systems – have not been defined. Also
the studies were done only at two stages – 15th and
30th day of vermicast generation. Hence, no useful
pointers can be drawn from these studies on the
impact of storage.
The present study, which is perhaps the first
of its kind, explores the changes in the physical,
chemical and biological properties of vermicast that
occur during storage with the objective of finding
conditions that minimize the deterioration in the
fertilizer value of the vermicast. The studies
provide useful pointers on how best to store and
package vermicast.
2. Materials and methods
2.1. Types of storage
The vermicompost used in the present work
was generated from paper waste and the epigeic
species, Eudrilus eugeniae. As paper waste is
almost entirely cellulosic, with only traces of
elements other than C, H, and O, the feed was
spiked with 9% w/w of cow dung in order to
provide NPK and other nutrients in adequate
amounts. The vermicomposting was accomplished
with a high–rate process recently developed by the
author’s group (Gajalakshmi et al., 2012). The
vermireactors were fabricated with aluminum
sheets and each had a volume of 135 liter (15 cm
height with surface area 150 x 60 cm). The
vermicast was harvested after 30 days. One part of
it was stored in two types of packs: (a) airtight
sealed transparent polyethylene bags of 20 micron
thickness (AUD), and (b) partially sealed nylon
mesh (0.3mm) bags (PUD). Both types of bags
were 25 cm long and 18 cm wide, each capable of
holding half kg of vermicompost. Another part of
the casts was pre-dried for 24 hours at room
temperature (29±4°C) and stored in both airtight
sealed transparent polyethylene bags (APD) and
partially sealed nylon mesh bags (PPD). In each set
36 packs were utilized; overall 144 packs were
studied. All storage was at room temperature
(29±4°C) as this is the temperature at which
vermicast is handled in the region where the
authors work. Three packs of vermicast were taken
once in a week for physical and biochemical
analysis from each storage.
2.2. Analysis
The physical properties of vermicast such as
bulk density, particle density and water holding
171
capacity (WHC) were determined by the standard
methods outlined by Bashour and Sayegh (2007)
and Margesin and Schinner (2005). The total
porosity and water filled pore space (WFPS) were
calculated from the particle and bulk density values
of the respective samples (Carter and Gregorich,
2008). Electrical conductivity (EC) and pH were
measured with sample suspension prepared with
distilled water (1:2, w/v) (Bashour and Sayegh,
2007). Total organic carbon (Corg) was determined
following modified dichromate redox method
(Heanes, 1984). Dissolved organic carbon (Cdis)
was extracted in 0.5 M K2SO4 solution (1:10, w/v)
and determined by the dichromate redox method
(Jenkinson et al., 2004). The total nitrogen (Ntot)
was determined by modified Kjeldahl method
(Kandeler, 1993) using Kel Plus™ semi-automated
digester and distillation units. Inorganic nitrogen
was extracted from moist samples with 2M KCl
solutions (1:10, w/v) followed by determination of
ammonium (NH4+-N) and nitrate (NO3
--N) content
in the suspensions by modified indophenol blue and
Devarda’s alloy method respectively (Jones, 2001).
Extractable potassium (Kext), calcium (Caext)
and sodium (Naext) were determined using a flame
photometer (Elico™ CL378) after extraction with
neutral 1N ammonium acetate solution. Extractable
phosphorus (Pext) was determined according to the
ammonium molybdate-ascorbic acid method
(Knudsen and Beegle, 1988) after extracting with
Mehlich 3 extraction solution (1:25, w/v) (Mehlich,
1984). Mineral sulfur (SO42-
-S) was extracted with
0.0125M CaCl2 solution (1:4, w/v), and determined
by turbidimetric method described by Bashour and
Sayegh (2007). Dehydrogenase enzyme activity
(DHA) was determined by iodo-nitro-
tetrazoliumchloride reduction method (Mersi and
Sehinner, 1991). β-glucosidase (BGA), alkaline
phosphatase (APA) and arylsulphatase (ASA)
enzymes activities were assayed by p-nitrophenol
method as described by Eivazi and Tabatabai
(1977, 1988) and Tabatabai and Bremner (1970).
Cellulase (CEA) activity was assayed by
determination of the reducing sugars released after
incubation of samples with carboxymethyl cellulose
sodium salt (Schinner and von Mersi, 1990). Urease
(URA) activity was assessed by incubating samples
with urea followed by determination of NH4+
released in the hydrolysis reaction by steam
distillation method (Tabatabai and Bremner, 1972).
Microbial biomass carbon (Cmic) was determined by
the chloroform fumigation-extraction method
(Jenkinson et al., 2004).
2.3. Processing of data
The experimental findings were statistically
analyzed to assess whether different treatments
exerted significant impact on the properties of
vermicompost over the course of the storage.
Pearson correlation was used to estimate the degree
of association between each of the vermicast
properties studied and their influence over others.
Statistical significance is recognized at p value ≤
0.05. The SPSS windows 16 package (Softonic,
Barcelona, Spain) was used throughout.
3. Results and discussion
3.1. Physical properties
The physical properties of vermicast were
significantly effected by pre-drying and storage
(Table 1). Pre-drying reduced the moisture content,
WHC, total porosity, and WFPS, while it increased
the bulk and particle densities of the cast (Figures
1, 2). In the course of 12 weeks, the moisture
content of the cast of PUD and PPD treatments was
reduced by 69.4±0.1 and 62.1±0.6%, and those of
the AUD and APD by 5.7±0.8 and 7.6±0.6%,
respectively. The bulk density increased in the PUD
and PPD storage to the extent of 49.7±0.8 and
45.8±1.7% and in AUD and APD to the extent of
21.8±2.0 and 29.1±0.3%, respectively. Structural
compactness occurring due to water loss in drying
172
Figure 1. Changes in the moisture content (a), bulk density (b), particle density (c) and water holding
capacity (d) of un-dried and pre-dried castings stored in airtight sealed bags (AUD and APD respectively)
and un-dried and pre-dried castings stored in partially sealed bags (PUD and PPD respectively), at different
periods of time.
may be the reason for the greater increase in the
bulk density in the PUD and PPD storage. The
WHC also reduced drastically due to this reason in
both PUD and PPD storage. It was reduced by
about 50% in PUD and PPD storage, in comparison
to 14.1±0.3 and 19.2±1.2%, respectively in the
AUD and APD storage. A significant linear
relationship (p<0.001) was found between WHC
and porosity of castings. At the end of the
experiments, the cast in the PUD and PPD storage
showed about 25% reduction in the total porosity
and those in the AUD and APD about 6% and 12%
reduction, respectively. Hydrophilic components of
organic matter, such as polysaccharides, might also
have influenced the WHC of castings (Li et al.,
2007).
The WFPS values increased to the extent of
22.4±3.4 and 32.7±0.5% in AUD and APD storage
and decreased by 18.4±2.0 and 8.5±5.8% in the
PUD and PPD storage, respectively. Reduction in
structural pores during dehydration and
decomposition of organic matter may be the reason
for lower WFPS of the PUD and PPD storage
0
20
40
60
80
0 2 4 6 8 10 12
Mo
istu
re c
on
ten
t %
No. of weeks
a
0.25
0.35
0.45
0.55
0.65
0 2 4 6 8 10 12
Bu
lk d
en
sity
g c
m-3
No. of weeks
b AUD APD
PUD PPD
1.05
1.10
1.15
1.20
1.25
0 2 4 6 8 10 12
Par
ticl
e d
en
sity
g c
m-3
No.of weeks
c
150
250
350
450
0 2 4 6 8 10 12
WH
C %
No.of weeks
d
173
Figure 2. Changes in the total porosity (a), water filled porosity (b), pH (c) and EC (d) of un-dried and pre-
dried castings stored in airtight sealed bags (AUD and APD respectively) and un-dried and pre-dried castings
stored in partially sealed bags (PUD and PPD respectively), at different periods of time.
(Schwärzel et al., 2002; Kechavarzi et al., 2010).
The changes in the WFPS of vermicast may have a
distinct impact on the regulation of hydrological
properties, gas diffusion, microbial colonization,
nutrient mineralization etc. (Gorres et al., 2001;
Schjønning et al., 2011; Yu et al., 2013). The
particle density of cast showed an increasing trend
in all the storage and the maximum of about 9%
increase was observed in the PUD and PPD. The
rate of increase in particle density of casts indicates
the degree of decomposition of organic matter they
contain (Hassink, 1995; Ruhlmann et al., 2006).
Increasing trend was also observed with EC, in
which maximum increase of 33.5±0.1 and
27.1±2.6% was in AUD and APD storage and
15.5±1.0 and 6.8±3.4%, respectively in PUD and
PPD. The castings in all types of storage had pH
close to neutral all the time (6.99 – 7.21); minor
fluctuations occurred due to the production of
organic acids and the release of CO2 during the
microbial decomposition of organic matter (Elvira
et al., 1998; Ahmad and Qazi, 2014).
3.2. Chemical properties
The chemical properties of cast were
significantly influenced by pre-drying and storage
55
65
75
85
0 2 4 6 8 10 12
Tota
l po
rosi
ty %
No. of weeks
a
15
19
23
27
31
0 2 4 6 8 10 12
WFP
S %
No. of weeks
b AUD APD
PUD PPD
6.9
7.0
7.1
7.2
7.3
0 2 4 6 8 10 12
pH
No.of weeks
c
1.0
1.2
1.4
1.6
0 2 4 6 8 10 12
EC m
mh
os
cm-1
No.of weeks
d
174
Table 1. F values of repeated measures analysis of variance on the effect of extend of sealing and pre-treatment on physical properties, EC and pH of vermicast
during the storage.
Treatment Moisture
content
Bulk
density Particle density
Water holding
capacity Total porosity
Water filled
pore space EC pH
Extend of Sealing 5676.8***
6641.1***
113.8***
886.9***
1781.0***
7.738* 5.290
n.s 4.503
n.s
Pre-Treatment 63025.5***
58466.9***
395.8***
8369.3***
17477.4***
6425.2***
2097.3***
0.318 n.s
Extend of Sealing X Pre-
Treatment 1627.7
*** 4155.3
*** 6.362
* 200.0
*** 1932.8
*** 329.73
*** 3.116
n.s 15.17
*
*p<0.05,
**p<0.01,
***p<0.001,
n.s - not significant.
Table 2. F values of repeated measures analysis of variance on the effect of extend of sealing and pre-treatment on chemical properties of vermicast during the
storage.
Treatment
Total
organic
carbon
Dissolved
organic
carbon
Total
nitrogen
Ammonium
-nitrogen
Nitrate-
nitrogen
Available
phosphorus
Exchangeable
potassium
Available
sulfur
Exchangeable
calcium
Exchangeable
sodium
Extend of Sealing 5.953* 10.94
* 1518.9
*** 2447.3
*** 336.5
*** 463.0
*** 271.6
*** 258.6
*** 6763.5
*** 2265.4
***
Pre-Treatment 2766.7***
2414.3***
38905.8***
16255.9***
113692.8***
775.4***
392.5***
1043.4***
1746.4***
34.74***
Extend of Sealing X Pre-
Treatment 0.346
n.s 19.59
** 1053.5
*** 104.3
*** 536.1
*** 36.84
*** 390.9
*** 791.8
*** 7676.5
*** 2464.6
***
*p<0.05,
**p<0.01,
***p<0.001,
n.s - not significant.
Table 3. F values of repeated measures analysis of variance on the effect of extend of sealing and pre-treatment on biochemical properties of vermicast during the
storage.
Treatment Dehydrogenase
activity
Cellulase
activity
β-Glucosidase
activity Urease activity
Alkaline
phosphatase
activity
Arylsulphatase
activity
Microbial
biomass carbon
Extend of Sealing 54.07
*** 19418.5
*** 1393.6
*** 11285.5
*** 4538.0
*** 2654.1
*** 1876.5
***
Pre-Treatment 98894.9
*** 13.06
** 12427.8
*** 4375.5
*** 5307.3
*** 65483.6
*** 4852.6
***
Extend of Sealing X Pre-
Treatment 544.3***
11275.7***
281.3***
3477.9***
3957.7***
760.1***
4.894 n.s
*p<0.05,
**p<0.01,
***p<0.001,
n.s - not significant.
175
(p < 0.001) (Table 2). Pre-drying reduced the Corg,
Cdis, Ntot, NH4+-N and Pext and increased the NO3
--
N, Kext, SO42-
-S, Caext and Naext content of the cast
significantly. The PUD and PPD storage showed
higher reduction in Corg, Cdis, Ntot, Pext and Kext than
the AUD and APD storage (Figure 3,4). In the case
of Corg, about 8% reduction was observed with
PUD and PPD storage and less than 1.5% in AUD
and APD. The higher reduction of Corg in the PUD
and PPD, which was maximum in the first two
weeks, was probably due to rapid mineralization of
C by aerobic microbes (Franzluebbers et al., 1994).
A few past studies on the ageing of cast of anecics
and endogeics have also reported high C
mineralization in the initial period (Martin, 1991;
Burtelow et al., 1998; Scullion et al., 2003). The
reduction in WFPS with time may be influencing
the extent of C mineralization as the days
progressed because reduction in the water-filled
pores during the storage might be curtailing the
microbial access to the substrate (Hassink et al.,
1993; Gorres et al., 2001). The cast in the PUD and
PPD storage also showed a maximum reduction in
Cdis content: more than 90% in comparison to
71.7±0.6 and 88.0±0.3%, respectively in the AUD
and APD storage. Low C utilizing efficiency of
anaerobic microbial community, which is expected
to dominate in the AUD and APD storage could be
the reason for the lesser reduction of Cdis content in
these types of storage (Søndergaard and Middelboe,
1995; Song et al., 2008).
The Ntot content of the cast reduced to the
extent of 66.9±0.5 and 54.6±0.3% in the PUD and
PPD storage, respectively. There was about 30%
reduction in the Ntot during the first week, probably
due to intense ammonia volatilization. The high
NH4+-N content of fresh castings also supports this
assumption. As the number of days progressed, the
ammonium content of the casts in the PUD and
PPD storage reduced up to 83% due to the intense
nitrification and ammonia volatilization from the
existing ammonium pools. There was a
concomitant increase in nitrate content of the cast
over time: 72.2±0.3 and 60.0±2.0% with the PUD
and PPD storage, respectively by the end of the
study. Previous studies on cast ageing have also
reported rapid exhaustion of most of the ammonium
present in the castings (Lavelle et al., 1992; Aira et
al., 2005; Kawaguchi et al., 2011b). In contrast, the
cast of AUD and APD storage showed reduction in
both NH4+-N and NO3
--N. Anoxic condition created
in AUD and APD storage due to the airtight sealing
might have impeded the nitrification process
(Jouquet et al., 2011; Aguilar et al., 2014), even as
the slow reduction in the mineral nitrogen content
of the casts may be due to microbial immobilization
(Lavelle and Martin, 1992; Decaens et al., 1999).
In all cases, the Pext in cast increased during
the initial week and further storage showed a steady
decline till the end. The high availability of carbon
and nitrogen in fresh cast would have increased the
phosphorus demand and it probably enhanced
phosphatase activity resulting in increased Pext
during the initial week (Parthasarathi and
Ranganathan, 1999; Flegel and Schrader, 2000). As
the number of days progressed, the Pext in the PUD
and PPD storage reduced by 71.5±0.3 and
73.9±1.5%. In AUD and APD it was reduced by
68.0±1.2 and 61.9±1.6%, respectively. The Kext in
the casts also increased in the initial week, but
further storage reduced it in all the treatments. This
was particularly pronounced in the PUD and PPD
when the reduction in Kext was 16.2±2.1 and
27.5±1.6%; in comparison to 5.4±2.0 and 1.6±2.3%
in AUD and APD, respectively. Similarly the SO42-
-
S content of the cast showed about 20% increase
during the initial week, while further storage
decreased it. The increase in the C/S ratio of the
cast to above 600 that occurred during this period
may be attributed to high immobilization of
available sulfur (Tabatabai and Chae, 1991; Reddy
et al., 2002). Throughout the experiment, the Caext
in the cast fluctuated in the AUD and APD storage.
Increased solubility of organic carbon and
increased competition between the cations for the
negatively charged sites due to increased levels of
Fe and Mn under reducing conditions may be the
reasons for this fluctuation (Wolt, 1994; Phillips
176
Figure 3. Changes in the total organic carbon (a), dissolved organic carbon (b), total nitrogen (c), ammonium
nitrogen (d), nitrate nitrogen (e) and available phosphorus (f) content of un-dried and pre-dried castings
stored in airtight sealed bags (AUD and APD respectively) and un-dried and pre-dried castings stored in
partially sealed bags (PUD and PPD respectively), at different periods of time.
250
260
270
280
290
0 2 4 6 8 10 12
Org
anic
car
bo
n m
g g-1
No. of weeks
a
0.0
2.0
4.0
6.0
8.0
0 2 4 6 8 10 12
DO
C m
g g-1
No. of weeks
b AUD APD
PUD PPD
3
6
9
12
15
0 2 4 6 8 10 12
Tota
l Nit
roge
n m
g g-1
No. of weeks
c
0
100
200
300
400
0 2 4 6 8 10 12
Am
mo
niu
m µ
g g-1
No. of weeks
d
0
100
200
300
0 2 4 6 8 10 12
Nit
rate
µg
g-1
No.of weeks
e
0
40
80
120
160
200
0 2 4 6 8 10 12
Ph
osp
ho
rus
µg
g-1
No.of weeks
f
177
Figure 4. Changes in the exchangeable form of potassium (a), sulfur (b), calcium (c) and sodium (d) content
of un-dried and pre-dried castings stored in airtight sealed bags (AUD and APD respectively) and un-dried
and pre-dried castings stored in partially sealed bags (PUD and PPD respectively), at different periods of
time.
and Greenway, 1998). The Caext in cast of PUD and
PPD storage steadily declined till the end, while
Naext declined to the extent of 11.1±0.8 and
31.1±0.1% in the PUD and PPD storage and
19.0±0.6 and 23.2±0.5%, respectively in AUD and
APD.
3.3. Biochemical properties
Pre-drying of the casts had strong influence
on the enzyme activities (Table 3). Except URA,
the activities of all other enzymes assayed – DHA,
CEA, BGA, APA and ASA – initially increased in
the pre-dried cast before declining. The extent of
this change varied with the type of storage (Figure
5). As much as 82 and 77% increase in DHA was
recorded in the first few weeks of AUD and APD
storage, but further storage reduced the DHA
activity to only 20.9±4.4 and 5.0±4.0%,
respectively. In the case of PUD and PPD storage,
DHA activity increased during the first week, and
then declined to the extent of 89.0±0.3 and
97.6±0.2%, respectively. This trend may be due to
enhanced growth of facultative anaerobic
microorganisms caused by exhaustion of oxygen in
10
12
14
16
18
0 2 4 6 8 10 12
Po
tass
ium
µg
g-1
No. of weeks
a
0
200
400
600
800
0 2 4 6 8 10 12
Sulf
ate
µg
g-1
No. of weeks
b AUD APD
PUD PPD
60
70
80
90
100
0 2 4 6 8 10 12
Cal
ciu
m µ
g g-1
No.of weeks
c
3.5
4.5
5.5
6.5
0 2 4 6 8 10 12
Sod
ium
µg
g-1
No.of weeks
d
178
Figure 5. Changes in the dehydrogenase (a), β- glucosidase (b), cellulase (c), urease (d), alkaline
phosphatase (e) and arylsulphatase (f) enzymes activity of un-dried and pre-dried castings stored in airtight
sealed bags (AUD and APD respectively) and un-dried and pre-dried castings stored in partially sealed bags
(PUD and PPD respectively), at different periods of time.
0
30
60
90
120
0 2 4 6 8 10 12
De
nh
ydro
gen
ase
µg
INT
g-1 2
h-1
No. of weeks
a
0
20
40
60
80
0 2 4 6 8 10 12
β-G
luco
sid
ase
act
ivit
y µ
g P
NG
g-1
h-1
No. of weeks
b AUD APD
PUD PPD
0
2
4
6
8
0 2 4 6 8 10 12
Ce
llula
se a
ctiv
ty µ
g C
MC
g-1
24
h-1
No. of weeks
c
0
40
80
120
160
200
0 2 4 6 8 10 12
Ure
ase
act
ivty
µg
NH
4-N
g-1
2 h
-1
No. of weeks
d
0
150
300
450
600
0 2 4 6 8 10 12
Ph
osp
hat
ase
act
ivit
y µ
g P
NP
g-1
h-1
No.of weeks
e
0
30
60
90
120
0 2 4 6 8 10 12
Ary
lsu
lph
atas
e µ
g P
NS
g-1 h
-1
No.of weeks
f
179
Figure 6. Changes in the microbial biomass carbon
content of un-dried and pre-dried castings stored in
airtight sealed bags (AUD and APD respectively)
and un-dried and pre-dried castings stored in
partially sealed bags (PUD and PPD respectively),
at different periods of time.
these types of storage (Stepniewska et al., 1990;
Brzezinâska et al., 1998). Further storage reduced
the DHA activity, possibly due to subsequent
decline in the availability of nutrients.
The BGA activity of the casts increased to
about 73% in the PUD and PPD storage by the
second week. Then there was a steady decline till it
fell to 48.8±0.2 and 49.5±1.2%, respectively at the
end. Similar trend was observed with AUD and
APD storage, in which there was about 67%
increase in BGA activity during the second week,
and 62.4±1.0 and 69.0±0.6% reduction by the
twelfth week. The CEA activity of castings also
increased with PUD, PPD and APD storage during
the initial week, but then fell as much as sevenfold;
those of AUD reduced ninefold. During the initial
week, more than four times increase in APA
activity was observed with the PUD and PPD
storage, and about twice with AUD and APD.
Further storage, reduced the APA activity in all the
cases. In the AUD and APD storage, the reduction
in APA activity was 40.8±1.1 and 47.1±1.2% and
in PUD and PPD storage it was 61.3±1.0 and
71.4±1.9%, respectively.
During the first week, there was a slight
increase in ASA activity with AUD and APD
storage. In the case of PUD and PPD storage, the
ASA activity increased up to third and fourth
weeks, respectively. Further storage showed a
drastic reduction in the ASA activity with all types
of storage: 93.3±0.4, 96.8±0.3, 97.9±0.2, and
99.4±0.1%, in AUD, APD, PUD and PPD,
respectively. Fresh casts had the highest URA
activity, which was reduced to 83.1±1.0, 89.7±0.3,
95.1±0.3, and 94.8±0.3% in AUD, APD, PUD and
PPD, respectively at the end.
In summary, except URA, the activities of all
other enzymes first rose in the initial weeks
possibly due to high availability of nutrients and
physical conditions favorable for aerobic microbial
growth (Allison et al., 2007). Subsequent decline in
the nutrient content (Sinsabaugh et al., 2005; Yao
et al., 2009), moisture content (Poll et al., 2006)
and availability of oxygen (Kang and Freeman,
1999; Xiao-Chang and Qin, 2006) with different
types of storage may have contributed to the
subsequent decline in the enzyme activities.
Pre-drying increased the Cmic content of the
casts significantly; it was 16.5±0.7% higher than
the fresh ones (Figure 6). During the first week,
Cmic increased to 20% in the PUD and PPD storage,
probably due to the high availability of nutrients
which might have promoted high microbial activity
during this period. Further storage led to reduction
across the board: 70.0±0.6, 78.2±0.7, 93.8±0.6,
96.3±0.2%, in AUD, APD, PUD, and PPD storage,
respectively. Subsequent decline in the availability
of nutrients and the moisture content in PUD and
PPD storage may probably be the reason for the
decline in Cmic. In the case of AUD and APD, the
reduction in Cmic may be attributed to the shift of
aerobic microbial groups to anaerobes due to
induced anoxic condition and which has very low C
0
5
10
15
20
0 2 4 6 8 10 12
Bio
mas
s ca
rbo
n m
g g-1
No.of weeks
AUD APD
PUD PPD
180
utilizing efficiency than the former (Song et al.,
2008).
4. Conclusions
A 3-month long study has been described on
the effect of storage on the fertilizer value of
vermicast. In what is arguably the first study of its
kind, several physical, chemical, and biological
attributes of the vermicast as stored with or without
pre-drying, and with or without airtight
containment, were assayed at 7–day intervals. The
manner of storage was seen to influence the plant-
friendly attributes of vermicast in a strong fashion.
Airtight storage after pre-drying was the most
beneficial, followed by airtight storage of the fresh,
undried, vermicast. In partially sealed storage there
was significantly more rapid deterioration of the
beneficial attributes than in airtight storage.
Interestingly, whereas 24-hr pre-drying before
airtight storage was helpful in retaining the plant-
friendly attributes of the vermicast for longer than
fresh-airtight storage, pre-drying before partially
sealed storage had the opposite effect. Apparently,
partially sealed storage added to the water loss that
had already occurred during the pre-drying, and
brought the water content below a level that was
needed to support biological activity within the
vermicast matrix. This indicates that a certain level
of water content is most appropriate for retaining
the microbiological and enzyme activities of the
vermicast; and the presence of water above or
below that level hastens the cast’s ageing. Further
work should be aimed at determining the most
beneficial water levels and how best to retain them.
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4:44
EFFECT OF PRE-DRYING AND EXTENT OF SEALING
ON THE PROPERTIES OF VERMICAST GENERATED
FROM THE NEEM LEAVES DURING STORAGE
Chapter
12
185
A paper based on this chapter has been
submitted for publication.
CChhaapptteerr 1122
Effect of pre-drying and extent of sealing on the properties of
vermicast generated from the neem leaves during storage
Abstract
In chapter 11, the effect of pre-drying and extent of sealing on the properties of vermicast generated from
paper waste was studied. To understand the impact of these storage practices on the properties of phytomass-
based vermicompost, a similar attempt has been made with the vermicast generated from the neem leaves. In
the present work, neem vermicast was packed in airtight and partially sealed bags with and without pre-
drying for 24 hours and the changes in the physical, chemical and biological characteristics of castings
during storage were monitored for 3 months at 7–day intervals. The storage of vermicast as explored in this
study showed significant differential influence on the properties of vermicast. Reduction in plant available
nutrient and enzyme activity along with changes in the physical properties of castings stored in partially
sealed bags indicate that their beneficial impact on plant growth may get reduced than the castings stored in
airtight bags. Even though, pre-drying of vermicast also reduced some of the beneficial properties of
vermicast during the initial period of storage, this process is recommended before storage as it has prevented
the structural disintegration of castings during storage.
1. Introduction
Enhanced physical, chemical properties and
microbial activity of soil with exogenous organic
amendment such as animal manure and compost
has been reported in numerous studies (Doan et al.,
2013; Ngo et al., 2013). In particular,
vermicompost has received renewed attention in
recent years due to its several plant-friendly
attributes. Application of vermicompost has shown
enhanced physical properties of soil such as
increase in water and nutrient storage capacity,
infiltration and aeration and resistance to
compaction and erosion (Edwards et al., 2004).
Since, most of the nutrients present in the vermicast
are in plant-available form in addition to the
presence of plant growth regulators such as auxins,
gibberellins, cytokinins produced by
microorganisms through interaction with
earthworm (Abbasi and Ramasamy, 1999; Atiyeh et
al., 2001) it possesses remarkable plant growth-
promoting potential on wide range of plants
(Gajalakshmi and Abbasi, 2004; Edwards, 2004).
Studies on a variety of crops such as cereals,
legumes, vegetables, ornamental, medicinal and
flowering plants assessed in both greenhouse and
field studies have confirmed that vermicast,
whether used as soil additives or as components of
plant growth container media, have significant
beneficial effects on seed germination, plant growth
and overall productivity (Edwards, 2004).
Development of disease resistance to a wide range
of plant pathogens is also reported with
vermicompost application (Sahni et al., 2008;
Szczech, 1999).
In the recent years due to the growing interest
of commercial sectors in vermicast, there is the
development of many large-scale commercial
vermicomposting units. In addition, many agro-
186
based industries throughout India, have adopted
vermicast production as part of their commercial
activities. Many thousands of tons of vermicast
produced from these units are supplied throughout
the country in packs which are either fully airtight
or partially sealed polyethylene bags. The vermicast
is packed after drying in shade or sometimes
without drying. There are no specified packing and
storage guidelines for vermicast. The physical,
chemical and biological properties of vermicompost
are subject to change during the storage which is
primary importance in the regulation of soil fertility
and plant growth promotion. Investigating the
effect of storage on vermicast is of great
importance with respect to understanding the
optimum age and storage condition for its best
utilization. The present work is an attempt towards
formulating packing guidelines for storing
vermicast.
2. Materials and methods
2.1. Experimental set up
Neem leaves were collected from the
Pondicherry University campus and its vicinity and
processed with Eudrilus eugeniae, an epigeic
earthworm species. After 30 days, the vermicast
was harvested. In one set of experiment, the
harvested fresh castings was stored in two types of
packing bags: (a) airtight sealed transparent
polyethylene bags of 20 micron thickness, and (b)
partially sealed nylon mesh (0.3mm) bags. The
second set of experiments comprised pre-dried
vermicast at room temperature for 24 hours and
stored in both types of packs. Both types of bags
were 25 cm long and 18 cm wide, each capable of
holding half kg of vermicompost. In this study,
there were 36 packs for each treatment, so totally
144 packs were prepared. All the packs of
vermicast were stored at room temperature in order
to imitate the general way of storage of
vermicompost in commercial sectors. Three packs
of vermicast from the all the treatments were
sampled once in a week for physical, chemical and
biochemical analysis.
2.2. Analytical methods
The analytical methods were the same as
detailed in section 2.2 of chapter 11.
2.3. Data analysis
Data was analyzed using repeated analysis of
variance at the 0.01% level for which with/without
pre-drying and type of sealing were fixed as
between-subject factors and storage period was
fixed as within-subject factor. Relationships
between different vermicast properties and their
influence over others were described and tested by
Pearson correlation coefficients. All analyses were
performed using SPSS 16 package.
3. Results and discussion
3.1. Physical properties
The change in the physical properties of
vermicast with different treatments is given in
Figures 1 and 2. During storage, castings stored in
partially sealed bags showed about 70% loss in
moisture content. In the airtight bags, a maximum
of 7.8% of water loss was recorded in 12 weeks of
storage. The changes in the moisture content of the
castings during air drying and storage has promoted
compactness in the structure resulting in the
increase in bulk density. An 11.5±1.6% increase in
bulk density was recorded during air-drying. The
bulk density of the castings stored in partially
sealed bags increased more sharply between the
first 42 days. At the end of 12 weeks, the increase
in bulk density was 52.0±0.2 and 45.3±1.9% with
un-dried and pre-dried castings, respectively. The
castings stored in airtight bags showed 21.6±3.8
and 25.9±2.6% increase in bulk density with un-
dried and pre-dried castings, respectively.
For organic matter, the particle density
depends on the degree of decomposition, and
187
Figure 1. Changes in the moisture content (a), bulk density (b), particle density (c) and water holding capacity (d), of
un-dried and pre-dried castings stored in airtight sealed bags (T1 and T2 respectively) and un-dried and pre-dried
castings stored in partially sealed bags (T3 and T4 respectively), at different periods of time.
ranges between 1.0 and 1.5 g cm-3
(Hassink, 1995;
Ruhlmann et al., 2006). In the present study, the
particle density of castings increased significantly
till the end of the experimental period irrespective
of the treatments. Increase in particle density was
higher in castings stored in partially sealed bags
than the airtight bags and it was ranging from 1.063
to 1.736 g cm-3
. In the case of water-holding
capacity (WHC) and total porosity, there was no
significant difference observed during the pre-
drying. The castings stored in the partially sealed
bags showed higher reduction of about 45% in
WHC in 12 weeks; whereas it was 19.0±0.8 and
34.3±1.2% with un-dried and pre-dried castings in
airtight bags. In this study, a significant linear
relationship (p<0.001) was found between WHC
and porosity of castings (Table 1). It indicates that
the decrease in WHC of castings during the storage
was likely due to the reduction of pore space in the
castings (Chaudhuri et al., 2009). Moreover, a
decrease in hydrophilic components of organic
matter e.g. polysaccharides (Piccolo and Mbagwu,
1999) during the storage might be limiting the
available surface area that absorbs water, resulting
in the decline in water holding capacity of castings
(Li et al., 2007).
The total porosity of castings decreased in all
the treatments. The castings stored in partially
sealed bags showed 10.3±0.5 and 11.8±1.0%
0
20
40
60
80
0 2 4 6 8 10 12
Mo
istu
re c
on
ten
t %
No. of weeks
a
0.25
0.35
0.45
0.55
0.65
0 2 4 6 8 10 12
Bu
lk d
en
sity
g c
m-3
No. of weeks
b T1 T2 T3 T4
0.9
1.1
1.3
1.5
1.7
1.9
0 2 4 6 8 10 12
Par
ticl
e d
en
sity
g c
m-3
No. of weeks
c
300
400
500
600
700
800
0 2 4 6 8 10 12
WH
C %
No. of weeks
d
188
Figure 2. Changes in the total porosity (a), water filled porosity (b), pH (c), and EC (d), of un-dried and pre-dried
castings stored in airtight sealed bags (T1 and T2 respectively) and un-dried and pre-dried castings stored in partially
sealed bags (T3 and T4 respectively), at different periods of time.
reduction in total porosity with un-dried and pre-
dried castings, respectively. The change in airtight
bags was only between 2.9±1.4 to 6.1±1.4%. The
water filled pore space (WFPS) of the castings was
reduced by 23.9±1.2% during pre-drying. In case of
airtight bags it was showing an increasing trend
with 20.7±6.4 and 24.8±4.3% increase in un-dried
and pre-dried vermicast respectively. In the
partially sealed bags, a steep reduction in WFPS
was observed as the number of days progressed and
it was 46.5±1.0% and 35.7±1.6% with un-dried and
pre-dried vermicast. Higher reduction in WFPS in
the partially sealed bags may be due to loss of
structural pores during dehydration and
decomposition (Schwärzel et al., 2002; Kechavarzi
et al., 2010). The changes in the WFPS of
vermicast have distinct impact on the other
physical, chemical and biological properties; WFPS
is of primary importance in the regulation of
hydrological properties, gas diffusion, microbial
colonization, nutrient mineralization etc. (Gorres et
al., 2001; Schjønning et al., 2011).
The pH of fresh vermicast was 6.71±0.01 on
the day of the deposition and it reduced to 6.11-
6.48 at the end of the experiment. The castings
stored in the airtight bags showed little fluctuation
in the pH, whereas the castings in partially sealed
bags exhibited a steady decline throughout the
experimental period. The reduction in pH during
63
67
71
75
79
0 2 4 6 8 10 12
Tota
l po
rosi
ty %
No. of weeks
a
10
20
30
40
0 2 4 6 8 10 12
WFP
S %
No. of weeks
b T1 T2
T3 T4
6.0
6.2
6.4
6.6
6.8
0 2 4 6 8 10 12
pH
No. of weeks
c
1.2
1.5
1.8
2.1
0 2 4 6 8 10 12
EC m
mh
os
cm -1
No. of weeks
d
189
Table 1. F values of repeated measures analysis of variance on the effect of extend of sealing and pre-treatment on physical properties, EC and pH of vermicast
during the storage.
Treatment Moisture
content
Bulk
density Particle density
Water holding
capacity Total porosity
Water filled
pore space EC pH
Extend of Sealing 1.420***
5.820***
2.487***
4.470***
72.95***
3.050***
111.1***
29.68**
Pre-Treatment 1.147***
5.372***
2.872***
1.534***
45.22***
871.6***
9913.9***
953.1***
Extend of Sealing X Pre-
Treatment 3.993
*** 497.1
*** 984.0
*** 687.7
*** 1.664
n.s 195.5
*** 672.5
*** 603.8
***
*p<0.05,
**p<0.01,
***p<0.001,
n.s - not significant.
Table 2. F values of repeated measures analysis of variance on the effect of extend of sealing and pre-treatment on chemical properties of vermicast during the
storage.
Treatment
Total
organic
carbon
Dissolved
organic
carbon
Total
nitrogen
Ammonium-
nitrogen
Nitrate-
nitrogen
Available
phosphorus
Exchangeable
potassium
Available
sulfur
Exchangeable
calcium
Exchangeable
sodium
Extend of Sealing 1.701***
1.576***
9.188***
3.363***
2.714***
257.2***
3.342***
9.410***
6.058***
1.114***
Pre-Treatment 4.671***
629.4***
2.363***
1.064***
1.067***
2.836***
141.0***
1.385***
35.85***
4.199***
Extend of Sealing X
Pre-Treatment 211.0
*** 532.4
*** 362.4
*** 4.556
*** 49.25
*** 13.71
** 622.2
*** 901.1
*** 888.3
*** 5.109
***
*p<0.05,
**p<0.01,
***p<0.001,
n.s - not significant.
Table 3. F values of repeated measures analysis of variance on the effect of extend of sealing and pre-treatment on biochemical properties of vermicast during the
storage.
Treatment Dehydrogenase
activity
Cellulase
activity
β-Glucosidase
activity Urease activity
Alkaline
phosphatase
activity
Arylsulphatase
activity
Microbial
biomass carbon
Extend of Sealing 9.370
*** 360.3
*** 3.454
*** 5.675
*** 3.341
*** 4.301
*** 2.209
***
Pre-Treatment 90.71
*** 1.930
*** 8.834
*** 1.893
*** 1.890
*** 9.446
*** 2.766
***
Extend of Sealing X Pre-
Treatment 155.3***
2.257***
4.569***
1.597 n.s
7.252***
8.230***
4.699***
*p<0.05,
**p<0.01,
***p<0.001,
n.s - not significant.
190
the storage may be due to production of carbon
dioxide and organic acid during the microbial
decomposition of organic matter (Elvira et al.,
1998). The electrical conductivity (EC) of fresh
vermicast was 1.44±0.03 mmhos cm-1
on the day of
deposition and it increased till the end of the
experiment with castings of airtight bags. In the
case of partially sealed bags, an increasing trend in
EC was observed during the first three weeks and
further storage reduced the EC till end of the
experimental period.
3.2. Chemical properties
There was a significant influence (p < 0.001)
on the chemical properties of vermicast with pre-
drying and storage (Figures 3 and 4; Table 2). Pre-
drying of vermicast showed reduction in organic
carbon and nitrogen content. During storage, the
castings stored in the partially sealed bags showed
constant reduction of C content and it was 6.5±0.1
and 7.3±0.2% for un-dried and pre-dried castings
respectively. The reduction of C content in partially
sealed bags was due to the rapid mineralization of
C by aerobic microbes in this treatment
(Franzluebbers et al., 1994). In this study, the C
loss was high during the initial week of storage and
this finding is similar to the report of Martin
(1991), Burtelow et al. (1998) and Scullion et al.
(2003). The reduction in C loss/mineralization as
the days progressed might be the reason for
reduction in WFPS which holds a greater
proportion of bacteria (Hassink et al., 1993). The
reduction in this pore size during the storage
curtails the microbial access to the substrate
(Gorres et al., 2001). The DOC in the vermicast
showed differential response to different treatments
(Figure 3b). The castings stored in partially sealed
bags showed a steady decline in DOC content
during the entire period of storage. In the course of
12 weeks storage, reduction in DOC was 86.0±0.2
and 88.4±0.1% with un-dried and pre-dried
castings, respectively. The castings stored in the
airtight bags showed maximum DOC content
throughout the experiment and the reason may be
the shift in aerobic condition in this treatment
which has reduced the C utilizing efficiency of the
microbial community (Søndergaard and Middelboe,
1995; Song et al., 2008). In this treatment, an
increasing trend in DOC content was observed
during second to fifth weeks, and further storage
decreased the DOC content by 44.3±0.8 and
37.6±1.0% with pre-dried and un-dried vermicast,
respectively.
The concentration of total nitrogen decreased
in all the treatments (Figure 3c) and the changes
were significantly different among the treatments
(p<0.001) (Table 2). The N concentration in un-
dried and pre-dried castings stored in partially
sealed bags reduced to 32.8±0.2 and 37.2±0.2% in
the course of 84 days storage. In these treatments
around 30% of nitrogen loss with pre-dried and un-
dried castings was observed in the first three weeks
of storage. The results reflected the high rate of
nitrogen loss in the castings during the initial weeks
of storage and it was likely due to the intense
ammonia volatilization. This result is in contrast to
the report of other field studies in which the total N
of castings were rather constant during the entire
ageing process (Decaëns et al., 1999; Jiménez and
Decaëns, 2004). These studies (Decaëns et al.,
1999; Jiménez and Decaëns, 2004) were conducted
with castings in the size of about 6 cm diameter
produced by the large anecic earthworm,
Martiodrilus sp. The larger size of the castings may
impede aeration and prevent the ammonium from
volatilization within the aggregates, whereas
castings that was stored in the present study was <
2 mm dia. Moreover, in the above cited studies the
pH of castings was 4.5 and 5 and the ammonia
volatilization completely stops at this pH (Hartung
and Phillips, 1994).
The concentration of NH4+ and NO3 in the
fresh castings was 669.4±3.9 and 57.5±0.9 mg kg-1
on the day of deposition. The high NH4+ content in
the castings may be due to high mineralization of
substrate during the gut passage and the addition of
urine in the posterior part of the digestive tract
191
Figure 3. Changes in the total organic carbon (a), dissolved organic carbon (b), total nitrogen (c), ammonium nitrogen
(d), nitrate nitrogen (e) and available phosphorus (f), of un-dried and pre-dried castings stored in airtight sealed bags (T1
and T2 respectively) and un-dried and pre-dried castings stored in partially sealed bags (T3 and T4 respectively), at
different periods of time.
340
360
380
400
0 2 4 6 8 10 12
Org
anic
car
bo
n m
g g-1
No. of weeks
a
0.0
1.0
2.0
3.0
4.0
5.0
0 2 4 6 8 10 12
DO
C m
g g-1
No. of weeks
b T1 T2 T3 T4
10
14
18
22
26
30
0 2 4 6 8 10 12
Tota
l nit
roge
n m
g g-1
No. of weeks
c
0
200
400
600
800
0 2 4 6 8 10 12
Am
mo
niu
m µ
g g-1
No. of weeks
d
0
200
400
600
0 2 4 6 8 10 12
Nit
rate
µg
g-1
No. of weeks
e
0
100
200
300
0 2 4 6 8 10 12
Ph
osp
ho
rus
µg
g-1
No. of weeks
f
192
Figure 4. Changes in the extractable potassium (a), sulfur (b), calcium (c) and sodium (d) of un-dried and pre-dried
castings stored in airtight sealed bags (T1 and T2 respectively) and un-dried and pre-dried castings stored in partially
sealed bags (T3 and T4 respectively), at different periods of time.
(Decaëns et al., 1999). Pre-drying of castings had
showed 35.5±0.2% of reduction in the initial NH4+
concentration. The decease of NH4+ in the castings
is reported to be reflected by increase in NO3-
concentration due to the nitrification of existing
ammonium pools (Lavelle and Martin, 1992;
McInerney and Bolger, 2000). Similar high
nitrification process was observed in the castings of
all the treatments during the first week of storage
and then it decreased progressively. Amongst
different treatments, maximum nitrification was
observed in castings stored in partially sealed bags.
In this treatment, the un-dried and pre-dried
castings showed 38.4±0.3 and 24.7±0.7% reduction
in NH4+ concentration at the first week, and then
81.7±0.7 and 78.5±0.5% reduction at the end of 84
days, respectively. The results are agreeing with
other findings of N dynamics during the cast
ageing. The castings of geophagous tropical
earthworm Pontoscolex corethrurus had reported
41% rapid nitrification during 16.5 days of
incubation (Lavelle et al., 1992). The casts of large
endogeic species Aporrectodea caliginosa has
shown up to 92.3% of NH4+ reduction during the 60
days laboratory incubation study (Aira et al., 2005).
Kawaguchi et al. (2011) reported that castings of
epigeic earthworm species Metaphire hilgendorfi
showed up to 30% NH4+ reduction during the 56
5
9
13
17
21
25
0 2 4 6 8 10 12
Po
tass
ium
µg
g-1
No. of weeks
a
0
200
400
600
800
0 2 4 6 8 10 12
Sulf
ate
µg
g-1
No. of weeks
b T1 T2
T3 T4
100
200
300
400
0 2 4 6 8 10 12
Cal
ciu
m µ
g g-1
No.of weeks
c
2.0
3.0
4.0
5.0
6.0
0 2 4 6 8 10 12
Sod
ium
µg
g-1
No.of weeks
d
193
days incubation.
The castings stored in airtight bags showed
30.2±0.8 and 35.1±0.5% of NH4+ reduction with
un-dried and pre-dried castings respectively. It can
be noted that the NH4+ reduction of up to
14.2±0.7% was observed in the first week itself and
then the rate of NH4+ loss largely reduced. Since,
nitrification process require oxic condition (Jouquet
et al., 2011), the anoxic condition created in airtight
bags after a week of storage would have impeded
this process. However, there was a slow reduction
in NH4+ observed throughout the storage process
along with gradual reduction in NO3-, certainly as
the result of denitrification and microbial
immobilization (Lavelle and Martin, 1992; Decaëns
et al., 1999). After 42 days of storage, fluctuation in
mineral N greatly reduced and this may be due to
reduction in microbial activity which in turn is due
to excess dryness. Similar findings were reported
with castings of endogeic earthworm Milsonia
anomala during the ageing process (Martin, 1991;
Lavelle and Martin, 1992).
In both airtight and partially bags increase in
labile P was observed in the initial week of the
storage and then there was a steady decline till the
end. Largely available nutrient pool and high
moisture content of fresh castings would have
enhanced phosphatase activity; the resultant higher
mineralization increased the fraction of labile P
pool in the vermicast during this period
(Parthasarathi and Ranganathan, 1999; Flegel and
Schrader, 2000). A similar result was also reported
with the castings of same earthworm species with
press-mud as feed and other species such as
Lampito mauritii and Martodrilus carimaguensis
during the ageing process (Satchell and Martin,
1984; Parthasarathi and Ranganathan, 1999;
Jimenez et al., 2003). Further storage decreased the
labile P content and there was no significant
difference between different treatments.
Exchangeable K content in the fresh castings
increased to 26.7±1.9% during the pre-drying and
this may be due to increase in the microbial
biomass during this period. Similarly, an increase in
K release during the drying was reported in soils
(Gupta And Rorison, 1974) and wet meadows
(Johnston et al., 1995). However, other authors
observed a decreased K release after soil drying
(Koerselman et al., 1993; Lamers et al., 1998), or
an unaffected K release (Grootjans et al., 1986).
During storage, the exchangeable K content
declined in all the treatments. The decrease in
exchangeable K was much higher in the casting of
partially sealed bags than the airtight bags (Figure
4a). Amongst all the treatments, the castings stored
in the partially sealed bags showed maximum of
43.5±0.6 and 67.6±0.3% reduction with un-dried
and pre-dried castings, respectively; whereas, the
decrease in airtight bags was 5.5±1.6 and
26.4±0.8% with the un-dried and pre-dried castings
during 84 days storage. The differential response of
exchangeable K during the storage with different
treatments could not be attributed to any particular
reason as relevant reports on the dynamics of K in
the vermicast and even on the biogeochemistry of
K in soils are very limited.
The sulfate content of vermicast increased by
7.8±1.1% during the pre-drying and the
mineralization of S continued to different extent
with different treatments (Figure 4b). The increase
in labile S pool in the vermicast during the pre-
drying was probably due to the breakdown of very
labile sulfate esters (Li et al., 2001). Increase in
arylsulphatase activity also indicates the increase in
microbial mineralization of sulphur during pre-
drying. During storage, an increase in labile S was
recorded in the initial weeks of storage with all the
treatments. Further storage showed reduction of
55.0±1.0 and 41.9±1.0% in S content with un-dried
and pre-dried castings stored in partially sealed
bags. In the case of airtight bags, the reduction was
about 30%. The castings stored in the airtight bags
showed fluctuation in both Ca and Na content
during the entire period of storage (Figures 4c and
4d). At the end of the experimental period, the un-
dried and pre-dried castings stored in airtight bags
showed 40.8±2.2 and 32.6±1.6% increase in Ca
194
content; whereas, in the partially sealed bags,
7.6±4.8 and 19.2±0.4% reduction in Ca content was
observed with un-dried and pre-dried castings,
respectively. The Na content of the un-dried
castings stored in the airtight bags showed increase
of 13.6±1.1% at twelfth week. Although, the pre-
dried castings showed a similar trend of results, at
the end of the experimental period about 30%
reduction in Na content was observed. In the case
of partially sealed bags, the Na content of the
castings was reduced about 50% during the storage.
The increase in Ca and Na content in the airtight
bags may be attributed to increased solubility of
organic carbon, and increased competition between
the cations for the negatively charged sites due to
increased levels of Fe and Mn under reducing
conditions (Wolt, 1994; Phillips and Greenway,
1998).
3.3. Biochemical properties
Typical changes of enzyme activity of
vermicast during the pre-drying and storage with
different treatments is shown in Figure 5. The
changes in enzyme activity were significantly
different among the treatments (p<0.001; Table 3).
It can be seen that dehydrogenase activity increased
to 22.4±0.4% during the pre-drying and the
increasing trend in this enzyme activity was
continued up to 42 and 49 days with un-dried and
pre-dried castings stored in airtight bags,
respectively. The increase in dehydrogenase
activity was more distinct in the un-dried castings
stored in airtight bags with the maximum of
74.4±0.3% higher activity in comparison to the
fresh castings. Whereas, 64.9±0.04% increase in
dehydrogenase activity was observed in the pre-
dried castings of similar treatment. After this, there
was a steady decline in dehydrogenase activity in
the airtight bags till the end of the experiment. The
castings stored in the partially sealed bags, showed
about 17% increase in dehydrogenase activity
during the first week and further storage decreased
the enzyme activity. In this treatment, about 85%
reduction in dehydrogenase activity was observed.
During storage, shift in aerobic to anaerobic
condition in airtight bags could be the reason for
the higher dehydrogenase activity (Brzezinâska et
al., 1998). Many studies on soil enzyme activity
reported similar high dehydrogenase activities in
anoxic condition (Stepniewska et al., 1990;
Brzezinâska et al., 1998).
The β-glucosidase activity increased to
34.1±0.5% during pre-drying. During the initial
period of storage, there was an increase in β-
glucosidase activity with both airtight and partially
sealed bags treatment, and the increase was more
distinct in the partially sealed bags. A maximum of
80% increase in enzyme activity was observed in
this treatment during the third week of storage;
whereas it was about 70% increase with airtight
bag. In the case of partially sealed bags, the higher
β-glucosidase activity during the first 3 weeks of
storage can be explained by their oxic condition.
Studies have shown similar changes in β-
glucosidase activity in the wetland soil during
changes in their oxic condition by varying moisture
content (Vo and Kang, 2013). From third week
onwards, a constantly reduced β-glucosidase
activity was observed in all the treatments till the
end of the experiment. Lower β-glucosidase activity
was observed in airtight bags indicating that the
change in the redox states of the soil impedes the β-
glucosidase activity in the vermicast; whereas, in
the partially sealed bag, reduction may be attributed
by reduction of microbial activity and degradation
of this enzyme due to excess dryness (Poll et al.,
2006).
Like other hydrolytic enzymes, the cellulase
activity also increased during the pre-drying
process. In this study, the fresh castings showed
1.34±0.02 µg CMC g-1
24h-1
cellulase activity and
increased by greater than twofold during pre-
drying. The un-dried and pre-dried castings stored
in the airtight bags showed a constant reduction in
cellulase activity and at the end of the twelfth week
reduction was 74.0±2.4 and 86.0±1.0%,
respectively (Figure 5c). Castings stored in the
195
Figure 5. Changes in the dehydrogenase (a), β- glucosidase (b), cellulase (c), urease (d), alkaline phosphatase (e) and
arylsulphatase (f) enzymes activity of un-dried and pre-dried castings stored in airtight sealed bags (T1 and T2
respectively) and un-dried and pre-dried castings stored in partially sealed bags (T3 and T4 respectively), at different
periods of time.
0
20
40
60
80
100
0 2 4 6 8 10 12
De
nh
ydro
gen
ase
µg
INT
g-1 2
h-1
No. of weeks
a
0
20
40
60
80
100
0 2 4 6 8 10 12
β-G
luco
sid
ase
act
ivit
y µ
g P
NG
g-1
h-1
No. of weeks
b T1 T2
T3 T4
0
1
2
3
4
0 2 4 6 8 10 12
Ce
llula
se a
ctiv
ty µ
g C
MC
g-1
24
h-1
No. of weeks
c
0
40
80
120
160
200
0 2 4 6 8 10 12
Ure
ase
act
ivty
µg
NH
4-N
g-1
2h
-1
No. of weeks
d
0
200
400
600
800
0 2 4 6 8 10 12
Ph
osp
hat
ase
act
ivit
y µ
g P
NP
g-1
h-1
No. of weeks
e
0
10
20
30
40
50
0 2 4 6 8 10 12
Ary
lsu
lph
atas
e a
ctiv
ity
µg
PN
S g-1
h-1
No. of weeks
f
196
partially sealed bags also showed a similar trend, in
which 75.3±1.4 and 89.1±1.3% reduction in
cellulase activity was observed with un-dried and
pre-dried castings, respectively. During storage, the
reduction in the cellulase activity of castings in all
the treatments may be probably due to reduction in
available nutrient such as inorganic N as reported in
soil and plant litter based studies (Sinsabaugh et al.,
2005; Yao et al., 2009).
The casting showed significantly high urease
activity of 180.6±2.2 µg NH4-N g-1
2h-1
on the day
of deposition and reduced to 8.5±1.3% during the
pre-drying. The declining trend of urease activity
was seen in all the treatments (Figure 5d). Amongst
different treatments, the un-dried and pre-dried
castings stored in partially sealed bags showed
fastest reduction in urease activity, where the
reduction was 50.0±0.5 and 54.2±0.4%,
respectively during the second week of storage. At
the end of the experiment, castings stored in these
treatments showed more than 22 and 26 folds
reduction in urease activity in the case of un-dried
and pre-dried castings, respectively. In the case of
castings stored in airtight bags, the reduction rate
was significantly lower than the castings of
partially sealed bags. In this treatment, urease
activity of un-dried and pre-dried castings reduced
to 84.1±0.4 and 88.2±0.3% respectively; it was
about 6 and 8 folds lower than the one day old
castings of respective treatments. The considerable
portion of urea present in the castings as earthworm
urine could be the reason of observed higher urease
activity in fresh vermicast (Edwards, 2004). The
decline in urease activity during the pre-drying and
storage probably happened due to the denaturing of
enzyme (Gould et al., 1973). Similarly, the change
in urease activity of soil during the incubation
under moist and dry condition has been reported in
previous studies (Zornoza et al., 2006; Geisseler et
al., 2011).
Alkaline phosphatase activity of the fresh
vermicast increased during the pre-drying and the
first week storage. As the days of storage
progressed, there was a steady decline in the
phosphatase activity with all the treatments. In the
case of un-dried castings stored in airtight bags,
decline in phosphatase activity was 44.7±1.6 and
36.5±0.3%, respectively; whereas in the partially
sealed bags, maximum reduction of 71.8±0.6% was
observed in the pre-dried castings followed by un-
dried castings which showed 45.4±0.9% reduction.
A possible explanation for the high phosphatase
activity during the pre-drying and first week of
storage may be due to high microbial activity in
vermicast during this period. Increase in labile N
and other nutrient content in the fresh vermicast
probably increased the P demand, a likely
consequence of higher phosphatase activity
stimulated by the active microbial population
(Allison and Vitousek, 2005; Allison et al., 2007).
The increase in labile P content by stimulated high
enzyme activity may have contributed to the slow
decline in phosphatase activity in the continuing
weeks of storage. In the case of castings stored
airtight, reduction in enzyme activity is probably
due to the low P requirement of anaerobic bacteria
and inhibition of phosphatase activity with oxygen
depletion as reported in many soil based studies
(Pulford and Tabatabai, 1988; Hinojosa et al.,
2004).
The castings showed 22.5±0.0 µg PNS g-1
h-1
of arylsulphatase activity on the day of deposition
and pre-drying increased the enzyme activity by
43.4±0.1%. Increase in arylsulphatase activity in
vermicast during the air-drying was consistent with
the reports of Tabatabai and Bremner (1970).
During storage, the castings stored in airtight bags
showed steady decline in enzyme activity from the
initial day of storage. In this treatment, 94.5±0.1
and 97.8±0.1% of reduction in enzyme activity was
observed within un-dried and pre-dried castings,
which was higher than the castings of partially
sealed bag treatments. The decline in arylsulphatase
activity in airtight bags is probably due to the
increase of redox states as reported on waterlogged
soil (Kang and Freeman, 1999; Xiao-Chang and
Qin, 2006). Reduction in arylsulphatase activity in
197
Figure 6. Changes in the microbial biomass carbon
content of un-dried and pre-dried castings stored in
airtight sealed bags (T1 and T2 respectively) and un-
dried and pre-dried castings stored in partially sealed
bags (T3 and T4 respectively), at different periods of
time.
anoxic condition might be the reason for changes in
the microbial community. Moreover, anaerobic
conditions also mobilize some metal ions, notably
Fe2+
, which might have impeded this enzyme
activity in castings stored in the airtight bags
(Pulford and Tabatabai, 1988; Freeman et al.,
1996). In the case of partially sealed bags, un-dried
and pre-dried castings showed increase in enzyme
activity during the initial period of storage, and then
there was a slow reduction in enzyme activity till
the end. In this treatment, a maximum reduction of
85.5±0.1% was observed in the pre-dried castings
followed by 67.1±0.1% in un-dried castings in the
course of 12 weeks of storage.
Microbial biomass carbon in the fresh
vermicast increased by 24.7±0.4% during pre-
drying. Storage of vermicast greatly affected the
microbial biomass C content of castings stored in
both airtight and partially sealed bags (Figure 6).
During the first week of storage, increase in the
microbial biomass C was observed in all the
treatments, and the increase was predominant in the
un-dried castings stored in both airtight and
partially sealed bags. During this period, the un-
dried castings showed 23.02±0.6 and 21.01±0.2%
increase in microbial C with partially sealed and
airtight bags, respectively. Continuing storage of
castings was characterized by reduction in
microbial C in the course of 12 weeks storage.
Castings stored in airtight bags showed 75.50±0.2
and 76.07±0.7% reduction in biomass with un-dried
and pre-dried castings, respectively at the end of the
experiment. Partially sealed bags treatment showed
steady decline in biomass and it was 85.03±0.5 and
94.05±0.2% with un-dried and pre-dried castings.
Although other studies showed reduction
(Parthasarathi and Ranganathan, 1999; Tiunov and
Scheu, 2000; Scullion et al., 2003) or no changes
(Aira et al., 2005, 2010) in biomass during the
initial period of ageing, in the present study higher
microbial activity and biomass was observed during
the first week.
Most of these previous studies are field based
and conducted with castings from the geophagous
earthworm, which are reported to contain a
considerable portion of mineral particles (Lavelle
and Spain, 2003; Blanchart et al., 2004; Jouquet et
al., 2008) and lower nutrient than the castings of
phytophagous worms, epigeic. This lower nutrient
content of the castings of geophagous are subject to
exhaustion very soon by intense microbial activity
(Tiunov and Scheu, 2000; Aira et al., 2005). In the
present study, availability of excess phytomass
attributed abundant nutrient availability in the fresh
vermicast, which would have created hotspot of
microbial activity during the first week of storage
and increased their biomass. The progressive
decline of available nutrients and moisture content
of vermicast was related to the resultant decline in
microbial biomass in further storage (Scheu, 1987).
The higher reduction of microbial C in the airtight
treatment could be due to the elimination of aerobic
microbial groups and shift in microbial community
due to induced anoxic condition. Microbial
communities under anoxic condition has low
energy yield from metabolizing reduced substrates,
leading to low C and other nutrient use efficiency
(Song et al., 2008).
0
5
10
15
20
25
0 2 4 6 8 10 12
Bio
mas
s ca
rbo
n m
g g-1
No.of weeks
T1 T2
T3 T4
198
4. Conclusions
The finding of this study reveals that
physical, chemical and biochemical properties of
vermicast are subject to modification during the
storage, irrespective of different treatments used in
this study. Amongst the treatments, the castings
stored in the partially sealed bags showed a drastic
change in their physical, chemical and enzymatic
properties. The physical properties such as
gravimetric water content, porosity and water-
holding capacity largely reduced whereas the bulk
density and particle density increased maximally in
the partially sealed treatment. These changes can
impede the water availability, oxygen diffusion and
plant root penetration in the field. Although, the
castings stored in airtight bags also showed changes
in these properties, it was less intense. The castings
stored in the partially sealed bags also showed a
higher reduction in total C, N and other nutrients
than the airtight treatment. Results of enzymes
assays and microbial biomass C content in partially
sealed treatment indicate intense aerobic microbial
activity in the initial period of storage which would
have immobilized and stabilized the available
nutrient pool in the organic matter.
Due to the excess dryness of castings stored
in partially sealed bag, nutrients would have highly
stabilized in organic matter and this would impede
the instant nutrient release to plants in the field.
Therefore, the beneficial impact of the vermicast on
the plant growth will be very minimum when stored
in partially sealed bags in comparison to either
fresh vermicast or castings stored in airtight bags.
Although, the beneficial properties reduced in the
castings stored in airtight bags, it was less intense.
Since there is no loss in nutrient due to their airtight
sealing and the physical properties of these castings
also favor the growth of microbes even after
prolonged storage, the disappeared nutrient pool
can be regained once it is applied into the field.
Even though, pre-drying of vermicast reduced some
of the beneficial properties of vermicast, this
practice can be recommended before storage as it
prevents the disintegration of the structure of
castings due to excess moisture. The small pellet
like structure of castings having higher surface area
will be available for microbial activity thereby
enhancing the mineralization of nutrients even after
prolonged storage.
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2134.
EFFECT OF PRE-DRYING AND EXTENT OF SEALING
ON THE PROPERTIES OF VERMICOMPOSTED COW
DUNG DURING STORAGE
Chapter
13
203
A paper based on this chapter has been
submitted for publication.
CChhaapptteerr 1133
Effect of pre-drying and extent of sealing on the properties of
vermicomposted cow dung during storage
Abstract
In chapters 11 and 12, studies on impact of pretreatment process and packing method on the properties of
vermicast generated from paper waste and neem leaves during storage were reported. In continuation to these
studies, the impact of the storage practices on manure based vermicast have been examined and reported in
this chapter. The changes in the physical, chemical and biochemical characteristics of castings during storage
were monitored for 3 months with weekly samples. The results reveal that the properties of castings get
altered irrespective of the mode of storage explored in this study. The castings stored in the bags which are
not fully airtight showed drastic losses in the plant available nutrient and enzyme activity along with adverse
changes in its physical properties than the castings stored in fully airtight bags. Even though, preprocessing
of vermicast such as drying has reduced some of the beneficial properties of vermicast; it prevented the
disintegration of the structure of castings due to excess moisture during storage.
1. Introduction
As discussed in chapters 11 and 12,
vermicast, whether used as soil additives or as
components of plant growth in container media,
have significant beneficial effects on seed
germination, plant growth and overall plant
productivity (Edwards, 2004; Lazcano, and
Domínguez, 2011). The beneficial aspects
contributed by vermicast on plant growth may be
attributed by various mechanisms, such as a
modification in soil structure, change in water
availability, increase in availability of macro and
micronutrients, stimulation of microbial and
enzymatic activities, or production of plant growth-
promoting materials by microorganisms through
interactions with earthworms (Abbasi and
Ramasamy, 1999; Atiyeh et al., 2001).
All these beneficial mechanisms rely on the
properties of vermicast which is subject to various
changes during storage. As there are no specified
packing and storage guidelines to preserve the
beneficial properties of vermicast during storage,
the present study was conducted. An an attempt has
been taken to explore the changes in the physical,
chemical and biological properties of manure based
vermicast during storage in order to understand the
optimum age and storage condition for its best
utilization.
2. Materials and methods
2.1. Experimental design
Vermicompost was generated from the cow
dung using an epigeic earthworm, Eudrilus
eugeniae. For vermicomposting, 140 litre volume
vermireactors fabricated with aluminum sheet were
employed. Fifty number adult worms per kg of feed
were introduced into 25 kg of cow dung on dry
weight basis and composted for 30 days. In order to
assess the impact of different storage method on
vermicast properties, the harvested vermicast was
stored by following methods: (i) fresh vermicast
stored in airtight bags (T1); (ii) vermicast pre-dried
204
for 24 hours (at room temperature under shade)
stored in airtight bags (T2); (iii) fresh vermicast
stored in partially sealed bags (T3), and (iv)
vermicast pre-dried for 24 hours stored in partially
sealed bags (T4). Each treatment comprised 36
packs of vermicast with 500 g of castings. All the
144 packs were stored at room temperature in order
to mimic the general way of storage of
vermicompost in commercial sectors. Three packs
of vermicast from the all the treatments were
sampled once in a week for physical and
biochemical analysis.
2.2. Analytical methods
The analytical methods were the same as
detailed in section 2.2 of chapter 11.
2.3. Data analysis
The influence of different storage methods on
the properties of vermicast were tested through
repeated analysis of variance. Pearson correlation
was also used to estimate the degree of association
between each of the vermicast properties studied
and their influence over others. All statistical
calculations were carried out using SPSS windows
16 package. Differences were considered
significant only when p-values were lower than
0.05.
3. Results and discussion
3.1. Physical properties
The physical properties of vermicast changed
significantly during the pre-drying and storage,
irrespective of different treatments used in this
study (Table 1). Pre-drying of vermicast reduced
the moisture content, water-holding capacity
(WHC) and water filled pore space (WFPS) by
11.2±1.3, 18.8±2.7, 11.48±1.42%, respectively;
whereas, the bulk density, particle density and total
porosity of the casting increased by 0.78±0.39,
2.66±0.19 and 0.60±0.18% respectively (Figures 1
and 2). As number of days progressed, there was a
steady decline in the moisture content, WHC, total
porosity and WFPS with all the treatments, and it
was more pronounced in castings stored in partially
sealed bags. In the partially sealed bags,
approximately 65% of moisture loss was recorded
during the 12 weeks of storage. The castings stored
in the airtight bags showed about 8% moisture loss
probably due to evaporation during sample
preparation.
During storage, the rapid change in the
moisture content of the castings stored in partially
sealed bags reduced the structural pores of organic
matter resulting in the increase in bulk density. In
this treatment, the bulk density increased more
sharply between the first to sixth week and in the
remaining period the changes was slow. The bulk
density of castings increased about twofold in
partially sealed bags. In this treatment, changes in
the bulk density of castings positively correlated
(p<0.001) with gravimetric water loss and WFPS in
the respective treatment (Table 1). The castings
stored in airtight sealed bags showed about 20%
increase in bulk density in the course of 12 weeks
of storage. Similarly, the particle density of castings
increased significantly in partially sealed bags than
the airtight bags. The particle density of castings
increased about 19 and 6% with partially sealed and
airtight bags, respectively. Changes in particle
density of castings depend on the degree of
decomposition, which reported to be varying
between 1.0 and 1.5 g cm-3
(Hassink, 1995;
Ruhlmann et al., 2006). During the storage, there
was no significant influence of pre-drying process
on particle density of the castings stored in both
partially sealed and airtight bags.
Total porosity of castings decreased around
20% in partially sealed bags, whereas it was less
than 6% with castings stored in airtight bags.
Nevertheless, the WFPS of castings showed a
different trend of results, which reduced maximally
in airtight sealed bags than castings of partially
sealed bags. The reduction in WFPS of castings
stored in airtight bags was 15 and 19% with un-
205
Figure 1. Changes in the moisture content (a), bulk density (b), particle density (c) and water holding capacity (d), of
un-dried and pre-dried castings stored in airtight sealed bags (T1 and T2 respectively) and un-dried and pre-dried
castings stored in partially sealed bags (T3 and T4 respectively), at different periods of time.
dried and pre-dried castings respectively. In the
partially sealed bags, 10 and 12% reduction in
WFPS was observed with un-dried and pre-dried
castings. During storage, the castings of partially
sealed bags developed low WFPS which may be
due to loss of structural pores during dehydration
and decomposition. This was reported with peat by
Schwärzel et al. (2002) and Kechavarzi et al.
(2010). The changes in the WFPS of vermicast
have a distinct impact on the other physical,
chemical and biological properties; WFPS is of
primary importance in the regulation of
hydrological properties, gas diffusion, microbial
colonization, nutrient mineralization etc. (Gorres et
al., 2001; Schjønning et al., 2011).
In general, water in the castings is retained in
pore spaces and adsorbed onto the organic matter
(Chaudhuri et al., 2009). In this study, a significant
linear relationship (p<0.001) was found between
water holding capacity and porosity of castings
(Table 1). In the partially sealed bags, WHC of the
castings reduced around 60% with both un-dried
and pre-dried castings, whereas it was 9 and 16%
with respective to airtight bag treatments. It
indicates that the decrease in WHC of castings
during the ageing process was likely due to the
20
30
40
50
60
70
80
0 2 4 6 8 10 12
Mo
istu
re c
on
ten
t %
No. of weeks
a
0.25
0.35
0.45
0.55
0.65
0 2 4 6 8 10 12
Bu
lk d
en
sity
g c
m-3
No. of weeks
b T1 T2 T3 T4
1.2
1.3
1.4
1.5
1.6
0 2 4 6 8 10 12
Par
ticl
e d
en
sity
g c
m-3
No.of weeks
c
100
200
300
400
500
0 2 4 6 8 10 12
WH
C %
No.of weeks
d
206
Figure 2. Changes in the total porosity (a), water filled porosity (b), pH (c) and EC (d), of un-dried and pre-dried
castings stored in airtight sealed bags (T1 and T2 respectively) and un-dried and pre-dried castings stored in partially
sealed bags (T3 and T4 respectively), at different periods time. reduction of pore space in the castings. Moreover, a
decrease in hydrophilic components of organic
matter e.g. polysaccharides (Piccolo and Mbagwu,
1999) during the storage might be restricting the
available surface area that absorbs water, resulting
in the decline in water holding capacity of castings
(Li et al., 2007).
The castings stored in both airtight and
partially sealed bags showed little reduction in the
pH throughout the experimental period. The
decrease in pH during the storage may be due to
CO2 and organic acid generated by the microbes
during the decomposition of organic matter (Elvira
et al., 1998). Moreover, the mineralization of
nitrogen and phosphorous into nitrites/nitrates and
orthophoshates and bioconversion of organic
material into intermediate species of organic acids
may also have contributed to lowering the pH of
vermicast (Ndegwa and Thompson, 2000; Suthar,
2007). The EC of castings stored in both airtight
and partially sealed bags showed increasing trend
and it was up to twofold higher than the fresh
castings.
3.2. Chemical properties
There was a significant influence (p<0.001)
55
60
65
70
75
80
0 2 4 6 8 10 12
`To
tal P
oro
sity
%
No. of weeks
a
20
24
28
32
36
0 2 4 6 8 10 12
WFP
S %
No. of weeks
b T1 T2 T3 T4
6.8
7.0
7.2
7.4
7.6
7.8
0 2 4 6 8 10 12
pH
No.of weeks
c
1
2
3
4
5
0 2 4 6 8 10 12
EC m
mm
ho
s cm
-1
No.of weeks
d
207
Figure 3. Changes in the total organic carbon (a), dissolved organic carbon (b), total nitrogen (c), ammonium nitrogen
(d), nitrate nitrogen (e) and available phosphorus (f), of un-dried and pre-dried castings stored in airtight sealed bags
(T1 and T2 respectively) and un-dried and pre-dried castings stored in partially sealed bags (T3 and T4 respectively), at
different periods of time.
300
320
340
360
0 2 4 6 8 10 12
Org
anic
Car
bo
n m
g g-1
No. of weeks
a
0
2
4
6
0 2 4 6 8 10 12
DO
C m
g g-1
No. of weeks
b T1 T2
T3 T4
10
20
30
40
0 2 4 6 8 10 12
Tota
l nit
roge
n m
g g-1
No. of weeks
c
0
200
400
600
0 2 4 6 8 10 12
Am
mo
niu
m µ
g g-1
No. of weeks
d
0
200
400
600
0 2 4 6 8 10 12
Nit
rate
µg
g-1
No.of weeks
e
0
100
200
300
0 2 4 6 8 10 12
Ph
osp
ho
rus
µg
g-1
No.of weeks
f
208
on chemical properties of vermicast with pre-drying
and storage (Table 2). Pre-drying of vermicast
showed reduction in organic carbon, DOC, total
nitrogen, NH4-N, extractable form of phosphorus,
potassium and calcium content of vermicast
(Figures 3 and 4). The NO3-N and sulfate content of
castings increased by 51.2±2.8, 13.5±1.0%,
respectively. The castings stored in the partially
sealed bags showed higher reduction of C content
than airtight bags and it was about 12% during the
12 weeks of storage; whereas, it was less than 3%
in castings stored in airtight bags. The higher
reduction of C content in partially sealed bags was
due to the rapid mineralization of C by aerobic
microbes (van Gestel et al., 1993) and the reduction
was maximum in the first two weeks. Few studies
on the ageing of castings of anecics and endogeics
have also reported high C reduction in the initial
period (Martin, 1991; Burtelow et al., 1998;
Scullion et al., 2003). The reduction in C
mineralization as the days progressed might be the
reason for reduction in WFPS which holds a greater
proportion of bacteria (Hassink et al., 1993). The
reduction in this pore size during the storage
curtails the microbial access to the substrate
(Gorres et al., 2001).
In the partially sealed bags, more than 86%
of DOC was exhausted in the 12 weeks storage. In
the case of castings stored in the airtight bags there
was increasing trend in DOC content during second
to fourth week and then there was a slow decline
till the end. However, the reduction rate in this
treatment was about half comparing to the partially
sealed bags. Less reduction of DOC in the airtight
bags may be due to the low C utilizing efficiency of
anaerobic microbial community (Søndergaard and
Middelboe, 1995; Song et al., 2008). The extent of
sealing of vermicast packs also showed significant
differential response to N loss from the vermicast.
The high aeration in the partially sealed bags would
have promoted the volatilization of ammonium
present in the fresh vermicast (Ndegwa and
Thompson, 2000; van der Stelt et al., 2007), hence,
there was around 25% loss of nitrogen during the
first two weeks of storage and the total N loss was
46% in 120 days.
The result of the present study is in contrast
to the report of ageing studies on vermicast in
which the total N of castings was rather constant
during the entire ageing process (Decaëns et al.,
1999; Jiménez and Decaëns, 2004). Studies
reported by Decaëns et al. (1999) and Jiménez and
Decaëns (2004) were conducted with castings of
size of up to 6 cm diameter and an average dry
weight of 25 g produced by the large anecic
earthworm, Martiodrilus sp. The larger size of the
castings may impede aeration and prevent the
ammonium from volatilization within the
aggregates, whereas castings stored in the present
study was < 2mm dia. Moreover, in the above cited
studies the pH of castings might be around 5 as the
pH of the study area was 4.5 and 5. This inference
is drawn based on the previous reports on Amynthas
khami, another anecic earthworm. The pH of fresh
castings was 4.3 to 5.7 where the pH of bulk soil
was in the range of 4.0 to 5.3 (Jouquet et al., 2008).
Generally, in different animal manure, large
ammonia volatilization takes place between a pH of
7 and 10: below pH 7 ammonia volatilization
decreases, and completely stops at pH of about 5
(Hartung and Phillips, 1994). Therefore, lower pH
of these castings may be the reason for no N loss
during the ageing. In addition, the diet of the above
mentioned anecic earthworm species consisted of a
considerable portion of mineral soil and the
castings they produced had a lesser concentration of
ammonium (220 to 290 µg g-1
) which was around
threefold lower than the castings used in the present
study.
The concentration of NH4+
-N in the fresh
castings was 525.6±8.2 mg kg-1
. The high NH4+
content in the fresh castings may be due to
mineralized nitrogen from the substrate during the
gut passage and the addition of urine in the
posterior part of the digestive tract (Decaëns et al.,
1999). The castings stored in partially
209
Table 1. F values of repeated measures analysis of variance on the effect of extend of sealing and pre-treatment on physical properties, EC and pH of vermicast
during the storage.
Treatment Moisture
content
Bulk
density
Particle
density
Water holding
capacity Total porosity
Water filled
pore space EC pH
Extend of Sealing 97.59***
0.257***
450.8***
484.3***
19.02***
57.91***
34.50***
459.3***
Pre-Treatment 3531.7***
11951.1***
9426.5***
1811.2***
5920.0***
195.6***
13889.2***
78.37***
Extend of Sealing X Pre-
Treatment 32.13
*** 23.63
*** 516.5
*** 110.9
*** 0.935
n.s 5.31
n.s 814.2
*** 73.76
***
*p<0.05,
**p<0.01,
***p<0.001,
n.s - not significant.
Table 2. F values of repeated measures analysis of variance on the effect of extend of sealing and pre-treatment on chemical properties of vermicast during the
storage
Treatment
Total
organic
carbon
Dissolved
organic
carbon
Total
nitrogen
Ammonium-
nitrogen
Nitrate-
nitrogen
Available
phosphorus
Exchangeable
potassium
Available
sulfur
Exchangeable
calcium
Exchangeable
sodium
Extend of Sealing 12.52***
86.57***
18.56***
7118.8***
191.4***
792.1***
36.44***
7679.8***
1014.7***
1165.1***
Pre-Treatment 313.0**
14198.0***
593.1**
26891.3***
23530.2***
1293.7***
10243.5***
660.1***
9912.2***
13756.7***
Extend of Sealing
X Pre-Treatment 13.94
** 10.35
* 12.49
** 442.4
*** 23.58
** 1.309
n.s 190.7
*** 2971.0
*** 374.2
*** 1758.1
***
*p<0.05,
**p<0.01,
***p<0.001,
n.s - not significant.
Table 3. F values of repeated measures analysis of variance on the effect of extend of sealing and pre-treatment on biochemical properties of vermicast during the
storage
Treatment Dehydrogenase
activity
Cellulase
activity
β-Glucosidase
activity Urease activity
Alkaline
phosphatase
activity
Arylsulphatase
activity
Microbial
biomass carbon
Extend of Sealing 2052.5
*** 703.6
*** 63.82
*** 903.1
*** 2311.4
*** 92.91
*** 18..15
**
Pre-Treatment 316645.8
*** 65.80
*** 11661.0
*** 1261.1
*** 76.24
*** 5681.6
*** 397.6
***
Extend of Sealing X Pre-
Treatment 2048.5***
1785.9***
1545.3***
3.706 n.s
3956.0***
7.259* 36.80
***
*p<0.05,
**p<0.01,
***p<0.001,
n.s - not significant.
210
Figure 4. Changes in the extractable potassium (a), sulfur (b), calcium (c) and sodium (d) of un-dried and pre-dried
castings stored in airtight sealed bags (T1 and T2 respectively) and un-dried and pre-dried castings stored in partially
sealed bags (T3 and T4 respectively), at different periods of time.
sealed bag showed a constant reduction in the NH4+
concentration and it was reflected by an increase in
NO3 concentration due to the nitrification of
existing ammonium pools (Lavelle and Martin,
1992; McInerney and Bolger, 2000). In this
treatment, about 21% of reduction in ammonium
content was observed in the first week, and then
there was more than 75% reduction at the end of 84
days. The results are agreeing with the findings on
N dynamics of castings generated from the
endogeic species Pontoscolex corethrurus,
Aporrectodea caliginosa and the epigeic Metaphire
hilgendorfi during the ageing process (Lavelle et
al., 1992; Aira et al., 2005; Kawaguchi et al.,
2011). In these studies, most of the ammonium
present in the vermicast was exhausted rapidly as
recorded in the present study. The castings stored in
airtight bags showed maximum NH4+ reduction of
18% during the first week of storage and then there
was a slow reduction till the end. The anoxic
condition created in airtight bags after a week of
storage would have impeded the nitrification
process (Jouquet et al., 2011). However, there was
slow reduction in NO3- observed throughout the
storage process along with gradual reduction in
NH4+, certainly as the result of denitrification and
microbial immobilization (Lavelle and Martin,
1992; Decaëns et al., 1999).
15
20
25
30
35
0 2 4 6 8 10 12
Po
tass
ium
µg
g-1
No. of weeks
a
0
200
400
600
800
0 2 4 6 8 10 12
Sulf
ate
µg
g-1
No. of weeks
b T1 T2
T3 T4
80
120
160
200
240
280
0 2 4 6 8 10 12
Cal
ciu
m µ
g g-1
No.of weeks
c
3.0
5.0
7.0
9.0
0 2 4 6 8 10 12
Sod
ium
µg
g-1
No.of weeks
d
211
Figure 5. Changes in the dehydrogenase (a), β- glucosidase (b), cellulase (c), urease (d), alkaline phosphatase (e) and
arylsulphatase (f) enzymes activity of un-dried and pre-dried castings stored in airtight sealed bags (T1 and T2
respectively) and un-dried and pre-dried castings stored in partially sealed bags (T3 and T4 respectively), at different
periods of time.
0
20
40
60
80
100
0 2 4 6 8 10 12
De
nh
ydro
gen
ase
µg
INT
g-1 2
h-1
No. of weeks
a
0
20
40
60
80
100
0 2 4 6 8 10 12
β-G
luco
sid
ase
act
ivit
y µ
g P
NG
g-1
h-1
No. of weeks
b T1 T2
T3 T4
0
1
2
3
4
0 2 4 6 8 10 12
Ce
llula
se a
ctiv
ty µ
g C
MC
g-1
24
h-1
No. of weeks
c
0
40
80
120
160
200
0 2 4 6 8 10 12
Ure
ase
act
ivty
µg
NH
4-N
g-1
2 h
-1
No. of weeks
d
0
200
400
600
800
0 2 4 6 8 10 12
Ph
osp
hat
ase
act
ivit
y µ
g P
NP
g-1
h-1
No.of weeks
e
0
20
40
60
80
0 2 4 6 8 10 12
Ary
lsu
lph
atas
e a
ctiv
ity
µg
PN
S g-1
h-1
No.of weeks
f
212
The available phosphorus content of the
castings stored in both airtight and partially sealed
bags increased by 20% in the un-dried castings, and
about 10% in the pre-dried castings during the first
week of storage, and then it progressively declined
over the 84 days of storage. In the initial weeks of
storage, largely available nutrient pool and high
moisture content enhanced phosphatase activity; the
resultant higher mineralization increased the
fraction of labile P pool in the vermicast
(Parthasarathi and Ranganathan, 1999; Flegel and
Schrader, 2000). The findings are very similar to
the reports on castings of Lampito mauritii,
Martiodrilus carimaguensis and other species
(Satchell and Martin, 1984; Parthasarathi and
Ranganathan, 1999; Jimenez et al., 2003). Further
storage, decreased the inorganic P content of
castings of partially sealed bags and the reason may
be due to reduction in microbial activity which in
turn is due to developing low moisture content.
Although similar trend of labile P content was
observed in the case of airtight bags, the amount of
labile P was higher than the partially sealed bags.
The reason may be due higher unutilized available
P pool due to reduction condition in this treatment
(Reddy et al., 2011).
During storage, the exchangeable K content
declined in all the treatments. The decreased
exchangeable K content in partially sealed bags was
about four times higher than the casting of airtight
bags. The amount of reduction in exchangeable K
was 12 and 46% with partially sealed and airtight
bags respectively. The variation in the response of
exchangeable K during storage with different
treatments could not be attributed to any particular
reason as relevant reports on dynamics of K in the
vermicast and even on the biogeochemistry of K in
soils are very limited. The sulfate content of the
vermicast stored in the partially sealed bags
increased by 25% during the first week of storage
and then there was a decline till the end. Increase in
arylsulphatase activity in this treatment indicates
the increase in microbial mineralization of sulphur
during this period. Further storage decreased the
sulfate content of vermicast and the reason may be
attributed to high C/S ratio of above 600 during this
period. The high C/S ratio might have promoted
the immobilization of available sulfur (Tabatabai
and Chae, 1991; Reddy et al., 2002). In the case of
airtight bags, except in the first week of storage, the
inorganic S was always higher than the castings of
partially sealed bags.
The castings stored in the airtight bags
showed fluctuation in both Ca and Na content
during the entire period of storage. In this
treatment, the maximum of 47% increase in Ca
content was observed during the tenth week of
storage with pre-dried castings. The Na content of
the pre-dried castings stored in the air tight bags
showed a maximum increase of 33% at the second
week followed by 17% with un-dried castings of
the same week. In the case of partially sealed bags,
both Ca and Na content steadily declined till the
end of experimental period. The increase in Ca and
Na content in the airtight bags may be attributed to
increased solubility of organic carbon, and
increased competition between the cations for the
negatively charged sites due to increased levels of
Fe and Mn under reducing conditions (Wolt, 1994;
Phillips and Greenway, 1998).
3.3. Biochemical properties
The changes in enzyme activity of vermicast
during the pre-drying and storage with different
treatments are presented in Figure 5. The β-
glucosidase, urease, alkaline phosphatase and
arylsulphatase activities of vermicast decreased by
14.6±0.4, 22.4±1.6, 11.3±4.9 and 14.8±2.9%
respectively during pre-drying. Whereas, the
dehydrogenase and cellulase activity increased by
1.77±2.63 and 29.1±2.6%, respectively. In the
airtight bags, the dehydrogenase activity increased
threefold with un-dried and pre-dried castings at
fourth and seventh week, respectively. After this,
there was a steady decline in dehydrogenase
activity till the end of the experiment. The rapid
exhaustion of oxygen present in the airtight bags
213
would have been bringing a shift of the activity
from aerobic to anaerobic microorganisms which
could be the reason for the higher dehydrogenase
activity (Brzezinâska et al., 1998). Many studies on
soil enzyme activity reported similar high
dehydrogenase activities in anoxic condition
(Glinski et al., 1986; Stepniewska et al., 1990;
Brzezinâska et al., 1998). The castings stored in the
partially sealed bags, showed a 19% increase in
dehydrogenase activity during the first week of
storage. After that there was a steady decline in
enzyme activity till the end of the experimental
period and it was about sixfold lower than the fresh
castings.
Unlike dehydrogenase, the β-glucosidase and
arylsulphatase activities of vermicast were higher in
the partially sealed bags than airtight bags. A
maximum of 74 and 77% increase in enzyme
activity was observed in un-dried and pre-dried
castings stored in partially sealed bags during the
third week of storage; whereas, it was 53 and 70%
in corresponding airtight bag treatments. At the end
of the experimental period, half of the initial
enzyme activity was reduced in both partially
sealed and airtight bags. The arylsulphatase activity
in partially sealed bags increased by 15 and 31%
with un-dried and pre-dried castings, respectively
during the initial period of storage, and then there
was a steady decline by 97% lower than the fresh
casting. The castings stored in airtight bags showed
steady decline in enzyme activity from the initial
day of storage and the maximum reduction of 97%
in the 12 weeks. The decline in arylsulphatase
activity in airtight bags is probably due to the
development of anoxic condition in this treatment
(Kang and Freeman, 1999; Xiao-Chang and Qin,
2006).
The cellulase, urease and alkaline
phosphatase activities of vermicast showed varying
response to different treatments. The cellulase
activity of pre-dried castings stored in airtight bags
and un-dried castings of partially sealed bags
increased by twofold during the first three weeks of
storage. Whereas, un-dried castings of the airtight
bags and pre-dried castings of partially sealed bags
showed 6 and 16% increase in cellulase activity
during the first week of the storage. The results
indicate that the castings which was exposed to
aeration either for short period during the pre-
drying or continuously in partially sealed bags has
increased the cellulase activity. The reduction in the
cellulase activity during further storage may be due
to the reduction in available inorganic N as reported
on soil and plant litter (Sinsabaugh et al., 2005;
Yao et al., 2009). The alkaline phosphatase activity
of vermicast increased by twofold in un-dried
castings of airtight bags and more than threefold
with others during the first week of storage. A
possible explanation for the high phosphatase
activity during the pre-drying and first week of
storage may be due to high microbial activity in
vermicast during this period. Increase in labile N
and other nutrient content in the fresh vermicast
probably increased the P demand, a likely
consequence of higher phosphatase activity
stimulated by the active microbial population
(Allison et al., 2007). The increase in labile P
content by stimulated high enzyme activity may
have contributed to the slow decline in phosphatase
activity in the continuing weeks of storage. A
maximum urease activity was observed in the fresh
castings probably due to a considerable portion of
urea present in the castings as earthworm urine
(Edwards, 2004). During the storage, the declining
trend of urease activity was seen in all the
treatments till the end of the experimental period.
The decline in urease activity during the storage
may be due to the reduction in urea content and
denaturing of enzyme (Gould et al., 1973). In this
experiment, about 6 and 15 times reduction in
urease activity was recorded at the end of 12 weeks
of storage.
Microbial biomass carbon in the fresh
vermicast increased by 12% during pre-drying. The
storage of vermicast greatly affected the microbial
biomass C content of castings stored in both airtight
and partially sealed bags (Figure 6). During the first
214
Figure 6. Changes in the microbial biomass carbon
content of un-dried and pre-dried castings stored in
airtight sealed bags (T1 and T2 respectively) and un-
dried and pre-dried castings stored in partially sealed
bags (T3 and T4 respectively), at different periods of
time.
week of storage, increase in the microbial biomass
C was observed in all the treatments, and the
increase was predominant in the castings stored in
partially sealed bags. Although other studies
showed reduction (Parthasarathi and Ranganathan,
1999; Tiunov and Scheu, 2000; Scullion et al.,
2003) or no changes (Aira et al., 2005, 2010) in
biomass during in the initial period of ageing, in the
present study, higher microbial activity and
biomass was observed during the first week. Most
of these previous studies are field based and
conducted with anecic and endogeic earthworm
species, which are reported to have a considerable
portion of mineral particles in their castings and
organic matter (Blanchart et al., 2004; Jouquet et
al., 2008) than the castings of epigeic species,
which is subject to exhaustion very soon by intense
microbial activity (Tiunov and Scheu, 2000; Aira et
al., 2005).
In the present study, castings generated from
the phytomass attributed abundant nutrient
availability in the fresh vermicast, which would
have created hotspot of microbial activity during
the first week of storage and increased their
biomass. Continuing storage of castings was
characterized by reduction in microbial C in the
course of 12 weeks storage. The progressive
decline of available nutrient and moisture content
of vermicast was related to the resultant decline in
microbial biomass in further storage (Scheu, 1987).
The higher reduction of microbial C in the airtight
treatment could be due to the elimination of aerobic
microbial groups and shift in microbial community
due to induced anoxic condition. Microbial
communities under anoxic condition has a low
energy yield from metabolizing reduced substrates,
leading to low C use efficiency. An unpublished
data reported by Song et al. (2008) shows that
microbial biomass C accounted for 2.28, 1.79, and
0.99% of soil organic C in oxic, intermittent, and
anoxic soils, respectively. The microbial
community structure with increasing oxygen
demand lead to the emergence of other stress
factors. For example, the nutritional stress indicator
in the anoxic soils was two to fourfold of that in the
oxic soils (Song et al., 2008).
4. Conclusions
The physical, chemical and biochemical
properties of vermicast were significantly altered
during the storage with different methods used in
this study. Amongst the treatment, the castings
stored in the partially sealed bags showed drastic
reduction in porosity, WHC and most of the plant
available nutrient studied. The bulk density of
castings also increased maximally in the castings
stored in partially sealed bags. These changes may
impede the water availability, oxygen diffusion and
plant root penetration and also low nutrient
availability in the vermicast when applied in field.
In addition, the excess dryness of castings stored in
partially sealed bags would have highly stabilized
the nutrients in organic matter and this would
impede the instant nutrient release to plants in the
field. The changes in the microbial and enzyme
activities of the castings support all the assumptions
mentioned above. Although, the beneficial
properties reduced in the castings stored in airtight
bags, it was less intense than the partially sealed
0
5
10
15
20
25
0 2 4 6 8 10 12
Bio
mas
s ca
rbo
n m
g g-1
No.of weeks
T1 T2 T3 T4
215
bags. Since the physical properties of these castings
also favor the growth of microbes even after
prolonged storage, the disappeared nutrient pool
can be regained once it is applied in the field. Even
though, pre-drying reduced some of the beneficial
properties of vermicast, this practice can be
recommended before storage as it prevents the
disintegration of the structure of castings due to
excess moisture; and would enhance the physical
properties and microbial activity.
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Biol. Biochem. 32, 265-275.
van der Stelt, B., Temminghoff, E.J.M., van Vliet,
P.C.J., van Riemsdijk, W.H., 2007.
Volatilization of ammonia from manure as
affected by manure additives, temperature and
mixing. Bioresour. Technol. 98, 3449-3455.
van Gestel, M., Merckx, R., Vlassak, K., 1993.
Microbial biomass responses to soil drying and
rewetting: The fate of fast- and slow growing
microorganisms in soils from different
climates. Soil Biol. Biochem. 25, 109-123.
Wolt, J.D., 1994. Soil solution chemistry:
Applications to environmental science and
agriculture. John Wiley and Sons, New York,
NY.
Xiao-Chang, W., Qin, L., 2006. Effect of
waterlogged and aerobic incubation on enzyme
activities in paddy soil. Pedosphere 16, 532-
539.
Yao, H., Bowman, D., Rufty, T., Shi, W., 2009.
Interactions between N fertilization, grass
clipping addition and pH in turf ecosystems:
Implications for soil enzyme activities and
organic matter decomposition. Soil Biol.
Biochem. 41, 1425-1432.
SUMMARY AND CONCLUSION
Chapter
14
219
CChhaapptteerr 1144
Summary and conclusion
Vermicomposting is the name given to the
process of conversion of biodegradable matter by
earthworms into vermicast. In general, vermicast
has been found to improve the seed germination,
growth, and productivity of many plant species. It
is also seen to improve soil health in terms of
aeration, water holding, and pH neutralization.
Although, there is accumulating scientific evidence
on these aspects, our understanding is largely
confined to the influence of vermicast derived from
the manure-based or manure-augmented substrates
on soils and plants whereas studies on vermicast
generated solely from phytomass are lacking. It has
also been a matter of concern whether phytomass-
based vermicompost will be as beneficial to the soil
and plants as manure-based vermicompost is. This
concern is particularly relevant for plants like
lantana and ipomoea which are known to possess
constituents that are toxic to animals and other
species of plants.
Information on the food preference and the
ageing of castings has been well documented for
geophagous and geophytophagous earthworms.
Much less attention has been paid towards
phytophagous earthworms. As vermicomposting is
primarily carried out using phytophagous species, it
is important to cover this knowledge gap as well.
The first chapter of the thesis comprises of
brief introduction to the studies reported in
Chapters 3 to 14. The importance of the studies
carried out by us in the context of the prior art is
brought out in Chapter 2.
Chapters 3-5 are dedicated to understanding
the feeding behavior of the epigeic earthworms,
which are species extensively used for
vermicomposting of various types of organic waste.
This group of earthworms is phytophagous in habit
and is believed to prefer organic matter, principally
phytomass. In contrast, the anecics and endogeics
consume an appreciable amount of mineral soil
with organic matter which facilitates assimilation of
nutrients in earthworm gut probably by keeping
their gizzard muscles toned up. To check whether
epigeics will ingest soil/sand even when there is
luxury availability of phytomass, set of experiments
were carried out with epigeic species, Eudrilus
eugeniae, and neem, ipomoea and cow dung as
feed.
It was seen that even though initially E.
eugeniae did ingest sand and soil despite the luxury
availability of phytomass, this tendency reduced as
the time passed indicating adaptive response to the
phytomass feed. Moreover, the assimilation of
soil/sand particles in the vermicast increased the
bulk density and particle density, reduced the pore
space, water holding capacity and nutrient content
of castings, which may reduce their beneficial
impact on plant growth and soil. These findings are
of significance in vermireactor’s design and
optimization because they indicate that sand-soil-
gravel bedding as used in conventional
vermireactors is necessary neither to ensure the
survival, growth and fecundity of the earthworm
nor the quality of the vermicompost.
Chapters 6-9 describe the effect of
vermicompost derived from the allelopathic weeds
such as lantana (Lantana camara) and ipomoea
(Ipomoea carnea), and paper waste on the
germination, growth and fruition of cluster bean
(Cyamopsis tetragonoloba). The impact of the
vermicompost (VC) was compared with that of an
220
inorganic fertilizer (IF) which had all the main
macro and micro-nutrients in concentrations
equivalent to the ones present in the VC. It was
seen that significantly greater germination rates
occurred in the lantana based VC treatments
compared to controls, while ipomoea based VC
showed suppression, indicating the presence of
some of the germination inhibitory components in
vermicast. However, both lantana and ipomoea
based VC also supported better plant growth in
terms of stem diameter, shoot length, shoot mass,
number of leaves, and leaf pigments. The positive
impact extended up to pod yield. In addition,
vermicast application enhanced root nodule
formation, reduced disease incidence, and limited
stunted plants.
The findings reveal that the allelopathic
ingredients of these weeds seem to have been
totally eliminated during the course of its
vermicomposting, and that these vermicompost has
the potential to support growth and fruit yield better
than equivalent quantity of inorganic fertilizers.
These studies open up the possibility that other
allelopathic weeds, and also plants which are toxic
in other ways, may be utilizable as substrates in
high-rate vermireactors as vermicomposting is
likely to destroy the toxic components of these
substrates as it is seen to have done in case of
lantana and ipomoea. This, in turn, may
enormously enhance the applicability of
vermicomposting as well as provide a means of
utilizing the biomass of several invasives which,
otherwise, goes to waste. In case of paper waste
based VC, application had no beneficial impact on
growth of cluster bean plant; whereas, the
corresponding IF treatments exhibited better seed
germination and plant growth than the former one.
In this experiment, the plants did not fructify in
both VC and IF treatments. The low fertile
experimental soil and insufficient minor and trace
nutrient present in the VC generated from the paper
waste might have impeded the production of
vegetables.
The findings of the studies reported in
Chapters 6-8 revealed that the vermicompost
derived from different parent materials have
different physical, chemical and biological
qualities, and their impact on germination, growth
and yield of plant also varied considerably.
Therefore, to understand the impact of
vermicompost from different parent materials on
soil health which is directly related to the growth
and yield of plants was assessed and reported in
Chapter 9. Several sets of experiments were carried
out in which the vermicast generated from different
organic wastes such as paper waste, leaves of
ipomoea, and of lantana or inorganic fertilizers
were applied in potting soil anchoring cluster bean.
Samples from all these treatments were collected on
weekly basis during different stages of plant
growth. The results reveal that vermicast
application created a conducive physical
environment by reducing bulk density, and
improving the water holding capacity and porosity
of soil. In addition, throughout the experiment, the
nutrient content of the soil was significantly higher
compared to inorganic fertilizers treated one.
Although, the inorganic fertilizers application
initially increased the nutrient content in soil, as the
days progressed, substantial quantity of applied
nutrient became unavailable to the plants. The loss
probably may be due to high leaching of mineral
nutrients from the soil. Apparently, no significant
impact has been observed on physical properties of
soil with inorganic fertilizers application.
Consequently, the vermicompost amendment may
be considered a good strategy for improving plant
growth, which reduces the deterioration of
agricultural lands due to the rampant use of
inorganic fertilizers.
The studies reported in Chapters 10-13,
which are perhaps the first of the kind, explores the
changes in the physical, chemical and biological
properties of vermicast that occur during storage
with the objective of finding conditions that
minimize the deterioration in the fertilizer value of
the vermicast. Vermicast generated from different
221
organic waste such as paper waste, neem and cow
dung was packed in airtight and partially sealed
bags with and without pre-drying for 24 hours.
Changes in several physical, chemical, and
biological properties of the castings were monitored
for three months with weekly assessments. The
manner of storage was seen to influence the plant-
friendly attributes of vermicast in a strong fashion.
Airtight storage after pre-drying was the most
beneficial, followed by airtight storage of the fresh,
undried, vermicast. In partially sealed storage there
was significantly more rapid deterioration of the
beneficial attributes than in airtight storage.
Interestingly, whereas 24-hr pre-drying before
airtight storage was helpful in retaining the plant-
friendly attributes of the vermicast for longer than
fresh-airtight storage, pre-drying before partially
sealed storage had the opposite effect. Apparently,
partially sealed storage added to the water loss that
had already occurred during the pre-drying, and
brought the water content below a level that was
needed to support biological activity within the
vermicast matrix. This indicates that a certain level
of water content is most appropriate for retaining
the microbiological and enzyme activities of the
vermicast; and the presence of water above or
below that level hastens the cast’s ageing. Further
work should be aimed at determining the most
beneficial water levels and how best to retain them.
In the present chapter, the gist of all the
studies reported in this thesis is given in brief.
222
STANDARDIZATION OF ANALYTICAL METHODS
Appendix
223
AAppppeennddiixx
Standardization of analytical methods
There are a number of analytical methods available for the variables assessed in the present study but in soil
or plant matrices. It is not known whether the methods will be applicable to vermicast also. Therefore, some
of the available analytical procedures for a few selected variables were identified on the basis of required
precision and accuracy. The procedures were then assessed for their applicability to vermicompost matrix by
means of the standard addition method, in which varying concentration of standards were spiked with
samples, and analyzed from its quantitative recovery. After ascertaining near-quantitative recovery with
adequate precision the methods were short-listed and utilized. The following table gives the details of the
standard addition methods for selected variables utilized for the experimental works detailed in the previous
chapters.
Parameters Methods Standards
Amount of
standard
added
Recovery
percentage
(Mean±SD) References
Total organic
carbon
Modified dichromate
redox method Glucose anhydrate 10-100 mg g
-1 94.5±2.4 Heanes, 1984
i
Total nitrogen Modified Kjeldahl method Ammonium nitrate 10-100 mg g-1
96.5±3.9 Kandeler,
1993ii
Ammonium
Extraction: Potassium
chloride extraction
Determination: Modified
indophenol blue method/
Nitroprusside catalyst
method
Ammonium sulfate 0.1-10 mg g-1
97.1±3.9 Bashour and
Sayegh, 2007iii
Nitrate
Extraction: Potassium
chloride extraction
Determination: Devarda’s
alloy method
Potassium nitrate 0.1-10 mg g-1
94.5±2.4
Gavlak et al.,
1994iv; Jones,
2001v
Extractable
Phosphorus
Extraction: Mehlich 3
extraction
Determination:
Ammonium molybdate-
ascorbic acid method
Potassium
dihydrogen
phosphate
0.1-10 mg g-1
96.8±3.6
Knudsen and
Beegle, 1988vi;
Mehlich,
1984vii
Extractable
Sulphate
Extraction: Calcium
chloride extraction
Determination: Turbidi-
metric method
Potassium sulfate 0.1-10 mg g-1
95.7±4.1
Houba et al.,
2000viii
;
Bashour and
Sayegh, 2007iii
Exchangeable
Potassium Extraction: Neutral
ammonium acetate
solution extraction
Determination: Flame
photometry
Potassium chloride 0.1-10 mg g-1
95.5±3.6
Lavkulich,
1981ix
Exchangeable
Calcium Calcium carbonate 0.1-10 mg g
-1 97.0±2.2
Exchangeable
Sodium Sodium chloride 0.1-10 mg g
-1 96.8±2.4
224
For each parameter, the Shewhart charts/control charts were prepared to assess the bias and precision
of standardization data (Figures 1-9). For that, the samples were fortified with known concentration of
chemical of interest, and quantitative recovery was calculated using following formula.
The quantitative recovery was plotted over the cumulative mean of the repeated measures. Control lines were
drawn which represent mean + 2 sigma (upper and lower warning limits, UWL and LWL) and mean + 3
sigma (upper and lower control limits, UCL and LCL). In these graphs, the warning lines (UWL and LWL)
represent a 95% confidence interval, and the control lines were corresponding to a 99% confidence interval
for acceptability of the methods for the specific matrix. In general, a single value outside the control lines is
considered unacceptable. The control charts for the few selected variables are shown in Figures 1-9. The
precision level and consistence in results for these variables are in acceptable range, hence these methods
have been utilized for analyzing the constituents in vermicast matrix.
Figure 1. Control chart of the quantitative recovery of organic carbon in vermicast matrix; where UCL – upper control
level, mean + 3σ; UWL – upper warning level, mean + 2σ; mean – average of values; LWL – lower warning level,
mean – 2σ; LCL – lower control level, mean – 3σ.
Figure 2. Control chart of the quantitative recovery of total nitrogen in vermicast matrix, where UCL – upper control
level, mean + 3σ; UWL – upper warning level, mean + 2σ; mean – average of values; LWL – lower warning level,
mean – 2σ; LCL – lower control level, mean – 3σ.
86
89
92
95
98
101
104
Sp
ike
reco
ver
y %
82
88
94
100
106
112
Sp
ike
reco
ver
y %
UWL
Mean
LWL
LCL
UCL
UWL
Mean
LWL
LCL
UCL
225
Figure 3. Control chart of the quantitative recovery of ammonium in vermicast matrix, where UCL – upper control
level, mean + 3σ; UWL – upper warning level, mean + 2σ; mean – average of values; LWL – lower warning level,
mean – 2σ; LCL – lower control level, mean – 3σ.
Figure 4. Control chart of the quantitative recovery of nitrate in vermicast matrix, where UCL – upper control level,
mean + 3σ; UWL – upper warning level, mean + 2σ; mean – average of values; LWL – lower warning level, mean –
2σ; LCL – lower control level, mean – 3σ.
Figure 5. Control chart of the quantitative recovery of extractable phosphorus in vermicast matrix, where UCL – upper
control level, mean + 3σ; UWL – upper warning level, mean + 2σ; mean – average of values; LWL – lower warning
level, mean – 2σ; LCL – lower control level, mean – 3σ.
83
88
93
98
103
108 S
pik
e re
cov
ery
%
86
90
94
98
102
106
Sp
ike
reco
ver
y %
83
88
93
98
103
108
Sp
ike
reco
ver
y %
UWL
Mean
LWL
LCL
UCL
UWL
Mean
LWL
LCL
UCL
UWL
Mean
LWL
LCL
UCL
226
Figure 6. Control chart of the quantitative recovery of extractable sulfur in vermicast matrix, where UCL – upper
control level, mean + 3σ; UWL – upper warning level, mean + 2σ; mean – average of values; LWL – lower warning
level, mean – 2σ; LCL – lower control level, mean – 3σ.
Figure 7. Control chart of the quantitative recovery of exchangeable potassium in vermicast matrix, where UCL – upper
level, mean + 3σ; UWL – upper warning level, mean + 2σ; mean – average of values; LWL – lower warning level,
mean – 2σ; LCL – lower control level, mean – 3σ.
Figure 8. Control chart of the quantitative recovery of exchangeable calcium in vermicast matrix, where UCL – upper
control level, mean + 3σ; UWL – upper warning level, mean + 2σ; mean – average of values; LWL – lower warning
level, mean – 2σ; LCL – lower control level, mean – 3σ.
83
88
93
98
103
108
Sp
ike
reco
ver
y %
83
88
93
98
103
108
Sp
ike
reco
ver
y %
89
92
95
98
101
104
Sp
ike
reco
ver
y %
UWL
Mean
LWL
LCL
UCL
UWL
Mean
LWL
LCL
UCL
UWL
Mean
LWL
LCL
UCL
227
Figure 9. Control chart of the quantitative recovery of exchangeable sodium in vermicast matrix, where UCL – upper
control level, mean + 3σ; UWL – upper warning level, mean + 2σ; mean – average of values; LWL – lower warning
level, mean – 2σ; LCL – lower control level, mean – 3σ.
i Heanes, D.L., 1984. Determination of total organic-C in soils by an improved chromic acid digestion and
spectrophotometric procedure. Commun. Soil Sci. Plant Anal. 15, 1191-1213. ii Kandeler, E., 1993. Bestimung von Gesamtstickstoff nach kjeldahl. In: Schinner, F., Kandeler, E., Ohlinger, R.,
Margesin, R. (eds.), Bodenbiologische Arbeitsmethoden, pp. 346-366. Spinger, Berlin. iii
Bashour, I., Sayegh, H.A., 2007. Methods of analysis for soils of arid and semi-arid regions. Food and Agriculture
Organization of the United Nations, Rome. iv Gavlak, R.G., Horneck, D.A., Miller, R.O., 1994. Plant, Soils, and Water Reference Methods for the Western Region,
Western Regional Extension Publication WREP 125, University of Alaska, Fairbanks, 45–47. v Jones, J.B., 2001. Laboratory guide for conducting soil tests and plant analysis. CRC Press, New York.
vi Knudsen, D., Beegle, D., 1988. Recommended phosphorus tests. In: Dahnke, W.C. (eds.), Recommended chemical
soil tests procedures for the North Central region, p. 12-15. Bulletin No. 499 (Revised), North Dakota Agric. Exp. Sta.,
Fargo, North Dakota. vii
Mehlich, A., 1984. Mehlich 3 soil test extractant: A modification of the Mehlich 2 extractant. Commun. Soil Sci.
Plant Anal. 15, 1409-1416. viii
Houba, V.J.G., Temminghoff, E.J.M., Gaikhorst, G.A., van Vark, W., 2000. Soil analysis procedures using 0.01 M
calcium chloride as extraction reagent. Commun. Soil Sci. Plant Anal. 31, 1299–1396. ix
Lavkulich, L.M. 1981. Methods Manual, Pedology Laboratory. University of British Columbia, Vancouver, British
Columbia, Canada.
88
91
94
97
100
103
Sp
ike
reco
ver
y %
UWL
Mean
LWL
LCL
UCL
228
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