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Available online at www.jpsscientificpublications.com
Life Science Archives LSA)
ISSN: 2454-1354
Volume 1; Issue - 2; Year 2015; Page: 112 - 123
2015 Published by JPS Scientific Publications Ltd. All rights reserved
Research Article
I n vitroSTUDIES OF ANTIFUNGAL AND ENZYMATIC ACTIVITY OF
SELECTED FUNGI ISOLATED FROM WATER USING DIFFERENT BAITS
S.A. Aghizion Inbakani1, V. Bhuvaneswari*
2, G. Kathiravan
3and B. Shanmugapriya
2
1S.D.N.B. Vaishnav College for Women, Chromepet, Chennai - 600 044, Tamil Nadu, India.
2Chikkaiah Naicker College, Veerapan chitram, Erode 600 004, Tamil Nadu, India.
3Department of Biotechnology ,Vels University, Pallavaram, Chennai600 117, Tamil Nadu, India.
Abstract
Aquatic fungi present in the fresh water ecosystem are of biological importance. Some serve as a
food for freshwater crustacean and other organisms. Some are parasitic on freshwater fishes while many aredestructive in aquarium and fish hatcheries. In this study, enumerations of fungi were carried out using moist
chamber incubation and baiting technique. Seeds, plants and animal segments, insects, fruits and vegetables
were used as baits. A total of 4614 fungal colonies were isolated. The relative percentage of the individualgroups of fungi revealed that hyphomycetes was maximum followed by zygomycetes, sachharomycetes,
ascomycetes, sterile morphospecies, coelomycetes and oomycetes. In addition to that the isolated fungi were
tested for enzyme activities such as amylase, laccase, and lipolytic activity. In vitro antifungal activity of
methanol extracts of some Indian medicinal plants against test fungi was done by agar disc diffusion methodto evaluate its potential importance. All the experimental test fungi subjected to enzyme assay showed
positive results for amylase activity whereas laccase activity was observed in only Trichoderma viride.Ou
of the methanol extracts of the five medicinal plants tested, Boerhavia diffusa, Lantana camara,andRicinus
communis showed best antifungal activity against Aspergillus flavus, Cladosporium cladosporioides, andDrechslera halodes. Thus, this work shows that the aquatic environment is blessed with abundant supply of
microorganisms waiting to be explored in various areas such as biodegradation, waste management etc.
ArticleHistoryReceived : 21.03.2015Revised : 01.04.2015
Accepted : 06.04.2015
Key words Aquatic fungi, Enzyme activityMedicinal plants and Plant extract.
1. Introduction
Fungi are universally present in all typesof natural waters and form one of the most
important components of an ecosystem as
decomposers. Mycelia frequently appear on seeds,fruits, petals, leaves, twigs and other elements of
plants fallen into water (Kiziewic, 2005). Baiting
techniques have provided a wealth of information
on isolation & distribution of aquatic fungi.
* Corresponding author: V. Bhuvaneswari
Tel.: +91-9176599550
E-mail: [email protected]
Several baits autoclaved wood cubes, filter paper
strips, dead fishes, meat piece, dead flies, insect
larvae, cooked egg white, and discs of cork havebeen used to isolate aquatic fungi (Alabi, 1971
Sharp 1978; Agina and Kpu, 1988). The use of
hemp seeds as baits (Sparrow, 1960; Lui and Volz1977; Sharp, 1978) for aquatic fungi is popular
Zoosporic fungi from different water bodies have
been studied in many parts of the world by
numerous researchers (Ziegler, 1958; Roberts1963; Alabi, 1971 a & b, 1974; El-Hissy, 1994)
Some fruits such as lemon, oranges, apples &
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pears are very good as baits for aquatic
phytophthora species (Adenle, 1994). The aim of
this study is to access the suitability of differentkinds of baits for the enumeration of fungi
associated with water and the effect of the
ecological factors including certain physical &
chemical parameters of water & composition ofthe substratum from which the fungi were isolated.
In addition to that the isolated fungi were tested
for enzyme activities such as amylase, laccase,
and lipolytic activity. In vitro antifungal activityof methanol extracts of some Indian medicinal
plants against test fungi was done by agar disc
diffusion method to evaluate its potentialimportance.
2. Methodology
General microbiological and laboratorytechniques followed in the present investigation
were as there outlined by Booth (1971). In thepresent investigation, four kinds of water samples
like pond, canal, lake and tap water were collected
from ten different locations. Water samples
employed in the study was collected in and aroundChennai, early in the morning usually between
5.00 a.m. and 7.00 a.m. and stored in polythene
bags. About 1000 ml water samples were collectedand used for further experimentation. The list of
location and kind of water samples are presentedin Table - 1.
Attempts were made to study and analyze
the physicochemical characteristics of the water(El-Hissy et al, 1990). The following parameters
were analyzed in Tamil Nadu Water Supply and
Drainage (TWAD) Board, State Level WaterTesting Laboratory, Chennai -
600 005. Physical examinations - Colour, odour,
turbidity, electrical conductivity, total dissolved
solids & suspended solids.Chemical examinations
- Alkalinity, pH, hardness, sodium, potassium,iron, manganese, ammonia, nitrate, nitrite,
chloride, fluoride, sulphate, phosphate, calciumcarbonate, and silica.
2.1. Methodology for recovery of fungi
associated with water samples
Isolation of fungi was carried out by using
the following isolation techniques viz.,
(a) Incubation and (b) baiting techniques in the
laboratory.
2.1.1. Incubation Method
In incubation method, small fragments of
substrates of plant decaying leaf litter, aquatic
plant parts, woody materials and animal origin(fish scales, gills, fins and fish tails) were
collected from the lake. The materials were broken
into small pieces and incubated on wet blotters inpetriplates. The materials along with petriplates
were kept in the incubator under laboratory
condition (222C temperature) for about 8 days(Czeczuga, 1991 a & b). At the end of the
incubation period, the colonized fungi on
incubated materials were examined
microscopically and the fungal hyphae were
transferred to a sterilized petriplate containingculture media. The following culture media
namely, Water Agar Medium (WAM), PotatoDextrose Agar (PDA) Medium, Corn Meal Agar
(CMA) Medium, Glucose Yeast (GY) Medium
and Glucose Yeast Peptone (GYP) Medium was
used for the enumeration of fungi.
2.1.2. Baiting Technique
Seeds were used as baits to isolate
zoosporic fungi (Farida et al., 2001; Kiziewicz
2005). The selected seeds were placed in one litrecontainers and covered properly to protect thewater from penetration by bacteria. After a few
days of incubation, microscopically determined
mycelia were removed from the seeds and
transferred to culture media amended withchloramphenicol and cycloheximide (150 mg/L)
to inhibit bacterial growth (Roberts, 1963). These
petridishes were examined periodically for
identification.
For the isolation of zoosporic fungi
associated with aquatic plant and animalsegments, samples were collected from the same
location and washed thoroughly under tap water toremove the debris and surface sterilized with 1%
mercuric chloride for 10 min and rinsed with
distilled water for about 5 min and then finallydried on a sterilized filter paper. The surface
sterilized segments were platted on culture media
for the enumeration of fungi.
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2.2. Methodology for recovery of fungi
associated with mud samples
The following baiting technique was used
for the recovery of fungi from mud samples (El-Hissy and Abal-Eloah, 1989). A 50 g of each mud
sample was introduced into clean sterilepetridishes. The mud sample in each petridish was
then flooded with sterile distilled water withaddition of few sesame seeds. The submerged mud
samples were also processed using certain
vegetables as bait. In this method, the selectedvegetable baits were swabbed with alcohol and a
hole of approximately 10 mm diameter was cut
through to the core on one side using a sterile cork
borer. The hole was packed with soil and coveredwith sticky tape to retain the soil. The baits were
incubated at room temperature in the light for 4 to
5 days. Isolations were made after the emergence
of mycelium from the baits.
2.3. Incubation, isolation and identification of
fungi
The petriplates were incubated in a light
chamber and observations were done from the
second day onwards for a period of 3 - 4 weeks forthe fungal colonies (Kiziewicz, 2005). The light
regime was 12 hours light followed by 12 hours
darkness. The hyphae, which grew out from the
bait samples were transferred to fresh PDA slants.They were maintained by sub-culturing. To
prevent the rapidly growing fungi from inhibiting
the slow growing species, the former wereremoved as soon as they appeared on the plates.
The identification of fungi was based on
morphological features such as shape and size ofhyphae, shape of sporangium and spores, structure
of oogonium, oosporum, and antheridium.
Identification and characterization of fungi were
made as per the key and standard monographs.
2.4. Analysis
Calculations were made in terms ofpercentage by using the following formula
2.4.1. Colonization frequency (CF %)
Individual fungal colonies that appeared in
the bait samples
CF % = 100
Total number of colonized segments
2.4.2. Relative percentage occurrence (RPO %)
The distribution and percentage ofoccurrence of different groups of fungi was
calculated by using the following formula.
Density of colonization of single group
RPO % = X 100
Total density of colonization
2.5. Enzyme activity of selected test fungi
The enzyme activity of the selected test
fungi were carried out according to the proceduregiven by Chamier, 1985).
2.5.1. Amylolytic activity
Glucose Yeast Peptone (GYP) medium
with 0.2% soluble starch with pH 6 was used
After 3 to 5 days of colony growth, the plates wereflooded with iodine solution. A yellow zone
around the fungal colony in an otherwise blue
medium indicated amylolytic activity.
2.5.2. Laccase activity
Glucose Yeast Peptone (GYP) medium
with 0.05 g, 1 naphtol/L with pH 6 was used. Asthe fungus grows, the colourless medium turned
blue due to the oxidation of 1 naphtol by laccase.
2.6. I n vitroantifungal activity of five medicinal
plants against test fungi
Fresh plant/plant parts were collectedrandomly from S.D.N.B. Vaishnav College
Chrompet, Chennai - 44. The details ofplant/plant parts screened, their familiesvernacular names and their therapeutic uses are
given in Table - 2. Fresh plant materials were
washed under running tap water, air dried and then
homogenized to fine powder and stored in airtightbottles. The air dried and powdered plant material
(10 g of each) was extracted with 100 ml of
methanol, kept on rotary shakes for 24 hrs and itwas filtered and centrifuged at 5000 rpm for 15
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min. The supernatant was collected and
evaporated to dryness to give the crude dried
extract which was used for anti fungal assay (JignaParekh and Sumitra Chanda, 2008).
2.7. Antifungal assay
To evaluate the antifungal activity, sterileagar plates were used according to the disc
diffusion assay. The test fungi were inoculated at
the centre of the agar plates. The sterile filterpaper discs (0.5 mm diameter) were impregnated
with plant extracts dissolved in methanol and
dried. Five individual discs of different plantextracts were placed around the inoculated test
fungi and incubated at 28C for 48 hrs. Following
an incubation period of 4 days, the plates were
removed from the incubator and antifungal
activity was evaluated by measuring zones ofinhibition of fungal growth. Clear zones within
which fungal growth was absent were measuredand recorded as the diameter (mm) of complete
growth inhibition. Blank disc impregnated with
solvent methanol followed by drying off was used
as control. The whole experiment was performedby making 0.5 cm wells on agar plates around the
test fungi and the wells were filled with 2 ml plant
extracts in DMSO (Dimethyl Sulphoxide) forcomparison with the disc diffusion method.
Table - 1: List of location and kind of water
sample collected during the course of study
. NoKind of
samplePlace of collection
1. Lake water Erumaiyur
2. Lake water Poonamallee
3. Canal Water Thiruneermalai
4. Canal Water Kundrathur
5. Pond Water Pallavaram
6. Pond Water Krishna Nagar
7. Pond Water Pazanthandalam
8. Pond Water Pallavaram
9. Tap WaterS.D.N.B. Vaishnav
College, Chrompet
10. Tap Water Pallavaram
Table - 2: List of selected medicinal plants and their therapeutic uses
S. No. Host plants Common Name Tamil Name Medicinal Uses
1. Melia azedarachL. Neem tree Vembu Anthelmintic, antifungal,antidiabetic, antibacterial,
antiviral, contraceptive and
sedative
2. Boerhavia diffusa
Linn.
Pular Punarnava Scabies, myalgia, aphrodisiac
3. Lantana camara
Linn.
Red Sage Mukkarattai Antipyretic, carminative,
Antidote to snake venom,treatment of malaria, wounds,cuts, ulcers, eczema, and tumours
4. Plumeria rubraLinn.
CommonFrangipani
Champige tree Ulcers, leprosy, inflammations,rube facient
5. Ricinus communis
Linn.
Castor oil plant Aamanakku Antimicrobial, antihistamine,
Anti-inflammatory treatment ofjaundice and sores.
http://en.wikipedia.org/wiki/Anthelmintichttp://en.wikipedia.org/wiki/Antidiabetichttp://en.wikipedia.org/wiki/Antibacterialhttp://en.wikipedia.org/wiki/Antiviral_drughttp://en.wikipedia.org/wiki/Contraceptivehttp://en.wikipedia.org/wiki/Sedativehttp://en.wikipedia.org/wiki/Antipyretichttp://en.wikipedia.org/wiki/Carminativehttp://en.wikipedia.org/wiki/Carminativehttp://en.wikipedia.org/wiki/Antipyretichttp://en.wikipedia.org/wiki/Sedativehttp://en.wikipedia.org/wiki/Contraceptivehttp://en.wikipedia.org/wiki/Antiviral_drughttp://en.wikipedia.org/wiki/Antibacterialhttp://en.wikipedia.org/wiki/Antidiabetichttp://en.wikipedia.org/wiki/Anthelmintic -
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3. Results and Discussion
The widespread occurrence of aquatic
fungi present in the freshwater ecosystem has been
gained recognition only rather recently. Thepresent study has been aimed towards isolation
and identification of fungi associated with tendifferent water samples collected from different
locations. Observations were made to variousparameters as mentioned in materials and
methods.
The hydrochemical analysis of ten
different water samples revealed that the lakewater samples of Poonamallee and Erumaiyur
(code no. 34659 and 34660) showed high level of
Ammonia. This indicates that the water is
extraneously polluted and the water sample is
chemically not potable. Similarly, the canal water(code no. 34657 and 34664) collected from
Thiruneermalai and Kundrathur showed high levelof ammonia and nitrate value, which exceeds the
maximum allowable limit and the water samples
are chemically not potable. Among the four pond
water samples analyzed, two samples fromErumaiyur and Pazanthandalam (code no. 34662
and 31872) was found to have more iron and
nitrate content and turbidity value exceeded themaximum allowable limit and the water samples
were chemically not potable whereas theremaining two pond samples (code no. 34663 and
34665) collected from Pallavaram andKrishnanagar and two more tap water samples
collected from S.D.N.B. Vaishnav college for
Women, Chromepet and Pallavaram were found tobe potable.
Further, the water analysis of ten samples
indicates that the pH range was between 6.9 7.9
and it has almost all the nutrients namely calcium,
magnesium, sodium, potassium, iron, manganese,
free ammonia, nitrate, nitrite, chloride, fluoride,sulfate and phosphate which supports growth of
the aquatic microorganism in natural environment.A total of 1456 fungal colonies were obtained
using seeds as bait. These colonies were classified
into 33 species of fungi belonging to 20 genera (1
Oomycetes, 2 Zygomycetes, 2 Ascomycetes and15 Hyphomycetes) and 13 non-sporulating sterile
morphospecies. The most frequently isolated
fungi using seeds as bait from ten experimental
water samples during the study period were
Pythium sp.,Mucorsp.,Rhizopussp, Chaetomium
globosum, Alternaria alternata, Cladosporiumcladosporioides, Fusarium oxysporum
Nigrospora sphaerica and Trichoderma viride
The total numbers of fungi recovered from theexperimental samples using seeds as bait are
presented in Figure 1a.
Of the 983 fungal colonies recovered, 20
taxa (2 Zygomycetes, 2 Ascomycetes and 14Hyphomycetes and 2 Coelomycetes) and 5 non-
sporulating sterile morphospecies were recorded
The commonly encountered fungi using plant
segments as bait during the study period wereChaetomium gracile, Alternaria alternate
Aspergillus flavus, Cladosporium cladosporioides
Geomyces sp., Monilia sp., Penicillium citrinum
Pestalotiopsis sp., and Colletotrichum sp. Thetotal numbers of fungal colonies from ten water
samples are presented in Figure 1b.
In this study of the 687 colonies recovered
16 genera (2 Zygomycetes, 2 Ascomycetes, 2Saccharomycetes and 10 Hyphomycetes) and 2
non-sporulating sterile morphospecies were
identified. The fungi namely Saccharomycopsissp., Brettanomyces sp., Acremonium
Paecilomyces sp. and Penicillium oxalicum weremore frequently isolated from the ten experimental
water samples using insects as bait. Theemergences of fungal colony from ten water
samples are presented in Figure 1c.
Altogether 450 fungal colonies (including
non-sporulating colonies) were recorded from theten experimental water samples using aquatic
animal segments as bait (Figure 1d). The
colonies were classified under 13 genera (2
Saccharomycetes, 1 Ascomycetes, 10
Hyphomycetes) and 2 non-sporulating sterilemorphospecies. The fungi such as
Saccharomycopsis, Acremoniumsp., Trichoderma
viride and Verticillium sp. were repeatedly
recovered during this study.
A total of 1038 fungal colonies belonging
to 22 genera were isolated from water samples
using fruits and vegetables as bait (Figure 1e)The fungal composition included 2 Zygomycetes,
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184
201
147
127
140
172160 164
60
101
0
50
100
150
200
250
Totalnum
beroffungalisolate
Lake water
sample 1
Lake water
sample 2
Canal water
sample 1
Canal water
sample 2
Pond water
sample 1
Pond water
sample 2
Pond water
sample 3
Pond water
sample 4
Tap water
sample 1
Tap water
sample 2
Total number of fungi recovered from the experimental samples using seeds as a bait
Sesame Pepper Chilli Cumin Green gram
2 Ascomycetes, 17 Hyphomycetes, 1
Coelomycetes and 5 non-sporulating sterile
morphospecies. The most frequently isolatedfungi from ten water samples were Mucor sp.,
Rhizopus stolonifer, Curvularia lunata,Fusarium
oxysporum, Geomyces sp., Humicola sp. and
Verticillium sp.
A total of 4614 fungal colonies wererecorded. These colonies were classified into 27
species of fungi belonging to 23 genera (1
oomycetes, 2 zygomycetes, 1 ascomycetes, 2saccharomycetes, 16 hyphomycetes and 2
coelomycetes) and the remaining 27 colonies were
classified under sterile morphospecies. The list of
fungi enumerated from water samples usingdifferent techniques are presented in Table - 3.
The ecological differences in differentgeographical locations play an important role in
the species diversity of the fungi. In the presentinvestigation, the relative percentage occurrence
of different groups of fungi from ten experimental
samples showed the percent contribution of
hyphomycetes was very high followed byzygomycetes, sachharomycetes, ascomycetes,
sterile morphospecies, coelomycetes and
oomycetes (Figure - 2).
A total of 1456 fungal colonies were
recorded from ten test samples using seeds as bait
The seeds differ in their effectiveness as baits forfungi associated with water. Among the five seed
baits tested seasame seeds trapped maximum
number of fungal colonies (361) followed by
pepper (304), chilli (285), cumin (277), and greengram (229). Some of the seeds belong to starchy
oily and proteinous food, hence their suitability as
bait may be due to their food content, which may
be comparable with the food requirements of thefungi isolated. The effectiveness may also be
attributed to the kind of seeds used as well as the
exposed nature of the endosperm of the seeds(Sparrow 1960).
Likewise, the total number of colonies
observed was maximum in plant twigs (225)
among the plant baits tested. Similarly, ant bait
trapped more fungal colonies (245) whencompared to the rest of the insect baits. Among the
animal segments tested for the bait fins showed
maximum number (131) of colonies. Out of thefruits and vegetable baits tested more number of
colonies were observed in carrot (236) followed
by apple, beetroot, lemon and potato.
Figure 1a: Total number of fungi recovered from the experimental samples using seeds as bait
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119
99
119 121
9295
129
97
5062
0
20
40
60
80
100
120
140
Totalnumberoffungalisolate
Lake water
sample 1
Lake water
sample 2
Canal water
sample 1
Canal water
sample 2
Pond water
sample 1
Pond water
sample 2
Pond water
sample 3
Pond water
sample 4
Tap water
sample 1
Tap water
sample 2
Total number of fungi recovered from the experimental samples using plant segments as a bait
Pla nt t wi gs Grass L ea f li tt er Nym phea pe tio le W oody ma te ria ls
91
114
69
86
6964
4943
55
47
0
20
40
60
80
100
120
Totalnumberoffungalisolates
Lake water
sample 1
Lake water
sample 2
Canal w ater
sample 1
Canal w ater
sample 2
Pond w ater
sample 1
Pond w ater
sample 2
Pond w ater
sample 3
Pond w ater
sample 4
Tap water
sample 1
Tap water
sample 2
Total number of fungi recovered from the experimental samples using insects as a bait
Ant Housefly Wings of insects
Figure 1b: Total number of fungi recovered from the experimental samples using
plant segments as bait
Figure 1c: Total number of fungi recovered from the experimental samples
using insects as bait
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2927
46
28
53
46
61
69
49
42
0
10
20
30
40
50
60
70
Totalnumberoffungalisolate
Lake water
sample 1
Lake water
sample 2
Canal water
sample 1
Canal water
sample 2
Pond water
sample 1
Pond water
sample 2
Pond water
sample 3
Pond water
sample 4
Tap water
sample 1
Tap water
sample 2
Total number of fungi recovered from the experimental samples using animal segments as a bait
Fish scales Gills Fins Fish tails
120114
129
125
90 91
134
117
56 62
0
20
40
60
80
100
120
140
Totalnumberoffungalisolate
Lake water
sample 1
Lake w ater
sample 2
Canal w ater
sample 1
Canal w ater
sample 2
Pond w ater
sample 1
Pond w ater
sample 2
Pond w ater
sample 3
Pond w ater
sample 4
Tap water
sample 1
Tap water
sample 2
Total number of fungi recovered from the experimental samples using fruits and vegetables as a bait
Apple Lemon Potato Carrot Beetroot
Oomycetes
Zygomycetes
Ascomycetes
Sachharomycetes
Hyphomycetes
Coelomycetes
SterileMorphospecies
Seeds
Plant segm ents
Insects
Animal segments
Fruits and vegetables
Percent contribution of different groups of fungi associated with water from
selected baits
Figure 1d: Total number of fungi recovered from the experimental samples
using animal segments as bait
Figure 1e: Total number of fungi recovered from the experimental samples
using fruits and vegetables as bait
Figure 2: Percent contribution of different groups of fungi
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Table - 3: List of fungi enumerated from the
water sample during the course of study
The most commonly encountered fungi invarious ecosystems namely Mucor sp., Rhizopus
sp., Chaetomium sp., Saccharomyces sp.,
Aspergillus sp., and Penicillium sp., have been
previously reported in the bathing sites of the
river, Poland (Bozena Kiziewicz, 2004). In thisstudy, the experimental baits such as aquatic plant
twigs, grass, woody materials and vegetables like
carrot, beetroot act as a good nutritive source for
the isolation of fungi like Humicola sp., Moniliasp., Acremonium sp., Gliocladium sp., Fusarium
sp., Colletotrichum sp., and Verticillium sp
Bozena Kiziewicz and Alicja Kurzatkowska 2004
have remarked that the fungi associated with thewater samples can be trapped by using animal
segments as bait. In this study, the genera
Saccharomycopsis and Brettanomyces wereisolated more frequently by using insects as bait
The fungi Pythium, which is reported here has
already been recorded as an aquatic fungi (Bozena
Kiziewicz, 2004).
Most of the fungal biomass on decayingleaves consists of vegetative hyphae that cannot be
identified through conventional microscopy
(Nikolcheva et al., 2003). Aquatic hyphomycetesan artificial phylogenitically, heterogenous group
of true fungi are fungi dominating leaf
decomposition in streams. They are anamorphs of
Ascomycota and Basidiomycota (Alexopoulos eal., 1996). The presence of sterile forms continues
to frustrate the mycologists because of their
uncertain taxonomy. Moreover, they demand theuse of molecular techniques for classification
(Bills, 1996). The nonsporulating sterile forms
recovered during the current study were separated
into culture groups based on the colonymorphology, hyphal mat characteristics (texture
zonation), presence of sclerotia and pigmentation
as described by Frohlich et al. (2000).
Apart from trying to understand thebiology of fungi associated with water another
motivation to study these fungi is its ability toproduce enzyme. Farida et al. (2001) reported
that, most of the aquatic fungi produced amylase,cellulase, pectinase, protease, lipase and xylanase.
These enzymes may cause breakdown of
leaf tissues and increase the palatability of the
leaves to leaf eating invertebrates (Barlocher1992; Suber Kropp, 1992). In the present
investigation, amylase activity was observed in
S. No. List of Isolated fungi
OOMYCETES
1 Pythiumsp.
ZYGOMYCETES
2 Mucorsp.
3 Rhizopus stolonifer
ASCOMYCETES
4 Chaetomium globosum
5 Chaetomium gracile
SACCHAROMYCETES
6 Saccharomyccessp.
7 Brettanomycessp.HYPHOMYCETES
8 Acremoniumsp.
9 Alternaria alternate
10 Aspergillus flavus
11 A. fumigates
12 A. niger
13 A. terreus
14 Cladosporium cladosporioides
15 Curvularia lunata16 Curvularia tuberculata
17 Drechslera halodes
18 Fusarium oxysporum
19 Geomyces sp.
20 Gliocladium roseum
21 Humicola sp.
22 Nigrospora sphaerica
23 Penicillium citrinum
24 Penicillium oxalicum25 Trichoderma viride
COELOMYCETES
26 Colletotrichumsp.
27 Pestalotiopsissp.
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almost all the test fungi. Among the 11 fungi
tested, one test fungi namely Trichoderma viride
showed that laccase activity which has beenpreviously reported (Zaldivar et al., 2001).
In general, plants generally produce many
secondary metobolites which constitute animportant source of microbicides, pesticides and
many pharmaceutical drugs (Satish et al., 2008;Vetrivel Rajan et al., 2009). Although most of the
fungi associated with water have been reported to
play an important role in decomposition andbioremediation, certain fungi can cause diseases in
plants and animals. Their spores infect plants
most frequently perforating the tissue that covers
stems, leaves, fruits and roots.
Aquatic and soil environments play a
significant part in the process of mycotic
infections. A similar study of screening naturaplant extracts against different fungal pathogens
was well recorded in literature (Banso and
Adeyemo, 2007). In this study, attempts were
made to investigate the inhibitory effects oforganic solvent extracts from 5 medicinal plant
species against 10 test fungi (Table - 4) to evaluate
the potential application of medicinal plant based
treatments to control disease caused by pathogenicfungi. Results revealed that extracts of Boerhavia
diffusa, Lantana camara and Ricinus communis
canbe used as a potent biocide to treat diseases inplants.
Table - 4: Antifungal activities of the test fungi
S.
No.
Antifungal activities of the test fungi
Name of the test fungi
Methanol extracts of selected medicinal plants
Zone of inhibition (mm)
Melia
azedarach
Boerhavia
diffusa
Lantana
camara
Plumeria
rubra
Ricinus
communis
1 Alternaria alternate 5 3 - 10 -
2 Aspergillus flavus 14 - 10 5 16
3Cladosporium
cladosporioides- 14 9 5 11
4 Curvularia lunata - 7 9 5 -
5 Drechslera halodes 11 9 7 - 8
6 Fusarium oxysporum - - - - 5
7 Penicillium oxalicum - 12 11 - 10
8 Pestalotiopsis sp. - - 12 - -
9 Rhizopus stolonifer - 3 5 - 4
10 Trichoderma viride - - - - 7
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2015 Published by JPS Scientific Publications Ltd. All rights reserved
4. References
1) Adenle. V. O. 1994. Comparison of techniques
for isolation aquatic phytophtora specis Niger.
J. Pl. Prot. 15: 1-9.2) Agina. S.E. and Kpu. R.S. 1988. A Survey of
aquatic Phycomycetes of rock fish farm in Jos,Plateau state Nigeria. Nigerian Journal of
Applied fisheries and Hydrobiology.3: 39-44.3) Alabi. R.O. 1971a. Factors affecting seasonal
occurrence of Saprolegniaceae in Nigeria.
Transaction of the British Mycological society.56: 289-299.
4) Alabi. R.O. 1971b. Seasonal periodicity of
Saprolegniaceae in Nigeria.Transaction of the
British Mycological society. 56:337-341.5) Alabi. R.O. 1974. Distribution of allomyces in
Nigeria soils. Transaction of the British
Mycological society. 63: 600-602.
6) Alexopoulous, C. J., C.W. Mims., and M.Blackwell. 1966. Introductory Mycology, 4
th
edition. Join Wiley and Sons, Inc. New York.
7) Banso, A., and S.O. Adeyemo. 2007.Evaluation of antibacterial properties of
tannins isolated from Dich rostachys cineria.
Afr. J. Biotechnol. 6 (15): 1785-1787.
8) Barlocher. R. 1992. The ecology of aquatichyphpmycetes springer-verlag, Berlin,
Germany.
9)
Bills, G.F. 1996. Isolation and analysis ofendophytic fungal communities from woody
plants. Endophytic fungi in grass and woody
plants Systematic, ecology, and evolution.
Edited by S.C. Redliss and L.M. Carris.American Phytopathological Society Press. St.
paul. Minn. Pp:31-65.
10)Booth, C. 1971. Introduction to general
methods In Methods in Microbiology. Vol:4.Ed. C. Booth Academic Press, London and
New York, pp 1-74.
11)
Bozena Kiziewicz. P.M., and M.K. AlicjaKurzatkowska. 2004. Aquatic fungi and
fungus like organisms isolated from surface
waters situated near Bialystok in podlasieprovince of Poland using Notonnecta glauca as
bait.Mycologi Balcanica. 117-123.
12)Bozena Kiziewicz, P.M. 2004. Water fungi
occurrence in the water reservoir in zarzeczany
of podlasie province. Wiladomosci
parazytologiczne. 50 (3): 587-593.
13)Chamier, C.A. 1985. Cell wall degradingenzymes of aquatic hyphomycetes: A review
Botany journal of Linnean Society. 91: 67-81.14)
Czeczuga, B. 1991a. Studies of aquatic fungi
part XXII. Mycoflora of the River Wegorapaand its Tributary, the River Goldapa Janka
Act a hydrochim Hydrobiol.19: 517-528.
15)Czeczuga, B. 1991b. Studies of aquatic fungiXVIII. Aquatic fungi in lake sniardwy and
eighteen neighboring lakes. Int. Revue.ges
Hydrobiol. 76: 121-135.
16)El-Hissy, F.T., A.M. Moharram, and S.A. El-zayal. 1990. Studies on the Mycoflora of
Aswan High Dam Lake, Egypt: Monthly
variation. Journal of Basic Mycrobiology.30
231-236.17)El-Hissy, F.T., and G.A. Abal Eloah. 1989
Aquatic fungi from Egyptian soils (upper
egy[pt). Sydowia.41: 150-159.18)El-Hissy. F.T. 1994. Oomycetes and
Chytridio Mycetes (Mastigo Mycetes) from
water bodies in Tubingen region (Germany)
J.Basic Microbiol.34: 67-75.19)Farida, T. El Hissy, S.M. Mortada, A.M
Khallil and F. Fatima, and A. Motual. 2001
Aquatic fungi recovered from water andsubmerged mud polluted with industrial
effluents. Online Journal of Biological
Sciences. 1(9): 854-858.
20)Forhlich, J., K.D. Hyde and O. Petrini. 2000Endophytic fungi associated with palms
Mycological Research. 104: 1202-1212.
21)Jigna Parekh and Sumitra Chanda. 2008. In
vitro antifungal activity of methanol extractsof some Indian Medicinal plants against
pathogenic Yeast and Moulds.African Journal
of Biotechnology. 7 (23): 4349-4353.22)Kiziewicz, 2005. Aquatic fungi growing on
seeds of plants in various types of water bodies
of podlaise province polosh. Journal ofEnvironmental Studies. 14 (1): 49-55.
23)Liu. C.H., and P.A. Volz. 1977. Freshwater
fungi of Taiwan.Mycologia. 69: 1079-1080.
24)Nikolcheva, L.G., a. Cockshutt and FBarlocher. 2003. Diversity of fresh water fungi
-
7/24/2019 16 LSA - Shanmuga priya New.pdf
12/12
V. Bhuvaneswari/Life Science Archives (LSA), Volume 1, Issue 2, Page 112 to 123, 2015 123
2015 Published by JPS Scientific Publications Ltd. All rights reserved
on decaying leaves comparing traditional and
molecular approach. Applied and
environmental microbiology. 69: 2548-2554.25)Roberts, R.E. 1963. A study of the
distribution of certain members of the
Saprolegnials. Trans Br. Mycol. Soc.46:213-
224.26)Satish.S., M.P. Raghavendra, M. P. Mohana,
.D. C., and K.A. Raveesha. 2008. Antifungal
activity of a known medicinal plant Mimusops
elengi L. against grain moulds. J. Agri.Technol. 4(1), 151-165.
27)Sharp. R.F. 1978. Investigative Mycology.
Heinemann Educational books Ltd. London.28)Sparroe. F.K. 1960. Aquatic Phycomycete 2
nd
Ed. Ann. Arbes. The University of Michigan
Press.
29)
Suber kropp.K. 1992. Interaction withinvertebrate. In.f. Barlocher (Ed). The ecology
of aquatic hypomycets. Springer-verlag,
Beclin, Germany. 118-134.30)Vetrivel Rajan. A, N. Shanmugavalli, C.
Greety Sunitha and V. Umashankar 2009.
Hepatoprotective effects of Cassia tora on
CCl4 induced liver damage in albino rats.IndianJ. Sci. Technol. 2 (3), 41-44.
31)Zaldivar, J., J. Nielsen, L. Olsson. 2001. Fuel
ethanol production from lignocelluloses a
challenge for metabolic engineering andprocess integration.Applied Microbiology and
Biotechnology. 56: 17-34.
32)Ziegler, A.M. 1958. The Saprolegniaceae ofFlorida.Mycologia. 50: 693-696.