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MANOHARBHAI PATEL INSTITUTE OF
ENGINEERING AND TECHNOLOGY
GONDIA
F O R W A R D I N G L E T T E R
Forwarded herewith to the Rashtrasant Tukdoji Maharaj Univesity, Nagpur, and the
dissertation
TREATMENT OF DOMASTIC WASTE WATER BY
PYTOREMEDATION (LEMNA PLANT)
Submitted by- Sandeep P. Ajmire , in partial fulfillment of the award of the
degree of Master of Technology in Environmental Engineering.
Prof. A. L. Nashine Prof. Dr.S.S. Rathor
Head of department Principal
Dept. of Civil Engg. MIET Gondia
MIET , Gondia
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MANOHARBHAI PATEL INSTITUTE OF
ENGINEERING AND TECHNOLOGY
GONDIA
C E R T I F I C A T E
This is to certify that dissertation entitled
TREATMENT OF DOMASTIC WASTE WATER BY
PHYTOREMEDATION (LEMNA)
Submitted by Sandip P.Ajmire , in practical fulfillment of the requirement for the
award of Degree of Master of Technology in Environmental Engineering to The
Rashtrasant Tukdoji Maharaj University, Nagpur , is bonafide research work carried
out under my supervision and guidance. The work embodied in this dissertation has
not submitted previously for the award of any degree or diploma.
Prof . A.M. Deshpande Prof.AL Nashine
Supervisor Head Of Department
Dept. Of Civil Engineering Dept. Of Civil Engineering
MIET GONDIA MIET GONDIA
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A C K N O W L E D G E M E N T
I express my profound gratitude towards Prof. A.M. Deshpande ,Lecturer ,
Department of Civil Engineering. MIET Gondia, for this able guidance.
I am extremely Grateful to Hon President Mr. Choudhari &
CEO A.V. Dhoke, Municipal Council Gadchiroli . Mr .M.G. Nisal , Lab Asst .
Environmental Engineering Lab MITE ,Gondia , without whose help the project
might have been completed. Mr. S.P. Waghmare, Executive Engineer Jeewan
Pradhikarn Gadchiroli & his technical and non technical staff , without whose help
the project might have been completed.
I express heartfelt thankful to Prof. Dr. S.S. Rathod , Principal & Prof
.A.L.Nashine , H.O.D., Civil Engineering & Prof. P.E.Mishra Coordinate, PG
Deptt. Of Environmentel Engg.,MIET, Gondia, for providing necessary facilities in
the completion of this work and for his constant encouragement.
Sandeep P. Ajmire
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C O N T E N T SINTRODUCTION
1.1. General
1.2 . Polution Problem
1.3 . Standards of Disposal
1.4. Treatment methodology
1.5. Objective and scope of study
LITERATURE REVIEW
2.1. General
2.2. Characteristics of domestic waste water
2.3. Tretment Processes
2.4. Process selection criteria for treatment of various domestic waste water
2.5. Application of Phytoremedation to domestic waste water
PHYTOREMEDATION
3.1. History & back round
3.2. Definition & types of Phytoremedation
3.3.Introduction of Phytoremedation by Lemna
3.3.1
3.3.2
3.3.3
3.3.4
3.3.5
3.3.6
3.3.7
3.3.8
3.3.9
3.4.
3.5
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PLANTS AND METHOD
4.1
4.2
4.3
4.5
4.6
4.7
4.8
4.9
4.9.1
4.9.2
4.9.3
4.9.4
4.9.5
4.9.6
4.9.7
4.10
OBSERVATIONS ,RESULTS,AND DISCUSSION
5.1
5.2
5.3
5.4
5.5
5.6
5.7
Reference:
PHOTOGRAPHS
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I N T R O D U C T I O N
1.1. General
The population of glob is increasing, the problem of municipal &
industrial waste tedious day by day. The legacy of rapid urbanization ,
industrialization ,fertilizer & pesticide use has resulted in major pollution
problems in both terrestrial and aquatic environments. In developing
countries is major problem to treat the polluted water from above sources.
Chemical & mechanical menace are used for this purpose is expensive. In
response , conventital, remediation systems based on high physical andchemical engineering approaches have been developed and applied to avert
or restore polluted sites. Much as these conventional remediation systems
are efficient, they are sparsely adopted because of some economical and
technical limitations. Generally, the cost of establishment and running deter
their use and meeting the demand particularly in countries with week
economy. Logical this high cost technology can neither be applied justifiably
where
1. The discharge is abruptly high for short time but the entire
average load is relatively small.
2. The discharge is very low but long term (entire load is medium).
3. The discharge is continuously decreasing over a long duration.
Thus conventional remediation approaches are best for circumstances
of high pollutants discharge like in industrial mining and domestic waste
water. Recently , it is evident that durability restoration and long term
contamination control in conventional remediation is questionable because
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in the long run the pollution problem is only is suspended or transferring
from one site to another.
The efficiency of duckweed (Lemna gibba L.) as an alternative cost
effective natural biological tool in wastewater treatment in general and
eliminating concentrations of both nutrients and soluble salts was examined
in an outdoor aquatic systems. Duckweed plants were inoculated into
primary treated sewage water systems (from the collector tank) for aquatic
treatment over eight days retention time period under local outdoor natural
conditions. Samples were taken below duckweed cover after every two days
to assess the plants efficiency in purifying sewage water from different
pollutants and to examine its effect on both phytoplankton and total and
fecal coliform bacteria.
The Lemnaceae family consists of four genera (Lemna, Spirodela,
Wolffia & Wolffiella) and 37 species have been identified so far. Compared
to most other plants, duckweed has low fiber content (about 5%), since it
does not require structural tissue to support leaves and stems. Of these,
applications ofLemna gibba L (duckweed) in wastewater treatment was
found to be very effective in the removal of nutrients, soluble salts, organic
matter, heavy metals and in eliminating suspended solids, algal abundance
and total and fecal coliform densities. Duckweed is a floating aquatic
macrophyte belonging to the botanical family Lemnaceae, which can be
found world-wide on the surface of nutrient rich fresh and brackish waters.
Outdoor experiments to evaluate the performance of the duckweed as a
purifier of domestic wastewater in shallow mini-ponds (20 & 30 cm deep)
showed that quality of resultant secondary effluents met irrigation reuse
criteria. Wastewater ammonia was converted into a protein rich biomass,
which could be used for animal feed or as soil fertilizer. The economic
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benefit of the biomass by-product reduced wastewater expenditures to
approx. US$ 0.05 per treated m3 of wastewater, which was in the range of
conventional treatment in oxidation ponds.
The present study was concerned with decreasing pollution of
municipal waste waster up to degree Standards of Disposal as per
National pollution control board.
1.2 . Pollution Problem
The quality of municipal wastewater of stagnant/ slow velocity may
create problem of high epidemics of malaria & other water born
diseases. The efficiency was tested by measuring some of
physicochemical characteristics of the control and plant treatments
after each seven days. The experiment lasted for a month, and recorded
the rates of reduction. The highest rates of reduction were for heavy
metals, accounting 99.8%, 99.6%, 98.7% and 72% for Copper,
Cadmium, Lead and Zinc, respectively, followed by Turbidity and
Nitrate recorded 64% and 57.1% respectively. The percentage
reduction of BOD5 was 49.6 %, while for COD 32.7%, either the rest of
the physicochemical characteristics as follows: 48.9% for Soluble Solids
and 43% of Oils and Grease, 41% of Total Alkalinity, 40% for Phenols,
39.1% for Sulfide, 38% of Suspended Solids and 30% of Phosphate.
Lower rates of reduction were recorded of each Temperature (17.2%),
pH and Sulfate (13.4%). The results showed that this aquatic plant canbe successfully used for wastewater pollutants removal
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Refinery wastewater is producing in a huge quantity in most the cities
of the country that contain a diverse range of pollutants including
Heavy Metals ,Oil and Grease ,Phenols, Sulphide, Sulphate ,Nitrate
,Phospate, Dissolved Solids, Suspended Solids, COD, BOD, which Its
disposal and treatment has become a challenge for the municipalities;
Many of the municipalities in growing cities neither have proper
disposal system nor have any treatment facility due to higher cost and in
such a situation refinery wastewater is discharge in to aquatic bodies
like river, ponds
and lakes, where it is posing a serious threat to the water quality and
become a big environmental problem throughout the development of
the petrochemical industry [1] [2].Physical, chemical, and biological technologies have been developed
to treat oily waste water and restore environmental quality; However
their costs are high and most of them are difficult to use under fieldconditions, hence in such a condition there is an urgent need to study
natural, simple, and cost-effective techniques for control pollution from
industrial effluents and treating such wastewater, such as
phytoremedation [1] [3].Viewing this fact Phytoremediation was assumed to be very useful, as
it is an innovative, eco-friendly and efficient technology in which naturalproperties of plant is used in engineered system to remediate hazardous
wastes through physical, chemical, and biological processes from
wastewater and sewage [4] [2].Phytoremedation is the utilization of plants accumulation capabilities
to remove contamination from water, soil and air, the capacity ofaquatic plants to remove pollutants from water is well documented [5].
The recent application of phytoremediation technology by duckweed inwastewater treatment and management is quite interesting andrevealing. Phytoremediation systems by duckweed are one of the
options that have been widely applied for combined handling of
wastewater with the nutrients used for poultry and aqua-culturalprojects [6] [7].
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Lemna minor L. known as common duckweed is a small, freefloating aquatic plant fast growing, adapt easily to various aquatic
conditions and play an important role in extraction and accumulation of
pollutants from waters [8]. In particular, species of Lemna are reportedto accumulate toxic metals and therefore are being used as experimental
model systems to investigate heavy metal induced responses,
Bioavaibility and bioaccumulation of various heavy metals in aquaticand wetland ecosystems is gaining tremendous significance globally [9].
Aquatic macrophytes take up metals from the water producing an
internal concentration several fold greater than their surroundings.Many of the aquatic macrophytes found to be the potential scavengers
of heavy metals from aquatic environment and are being used inwastewater renovation systems [10] [11]. Aquatic plants have shown
their efficiency in absorbing nutrients from various sources of polluted
water, [12] [13].This study aimed to assessing the efficiency of duckweed (Lemna minor)
in phytoremediate the pollutants of Basrah oil refinery wastewater.
Pour dterminer la tolrance et la capacit de phytoaccumulation ducuivre (Cu) et du nickel (Ni) par
une espce de lentilles deau, Lemna gibba L., les plantes sont exposes
diffrentes concentrationsde Cu et Ni (0,1 2,0 mg/L) dans une solution de Coc et Lesaint dilu
1/4. Le pH est maintenu
constant 6,0 ( 0,1) et le flux de lumire est de 12 h/jour. Le cuivre etle nickel sont tolrs par L.
gibba des concentrations 0,3 mg/L et 0,5 mg/L, respectivement.
Cependant, la croissance desplantes diminue de 50% (I50) quand le milieu de culture contient 0,45
mg/L de Cu ou 0,75 mg/L de Ni.La plus faible concentration causant une inhibition complte (LCI) est
de 0,5 et 1,0 mg/L
respectivement en prsence de Cu et Ni. Les rsultats de lanalyse dumtal dans les tissus des
plantes rvlent une grande accumulation de Cu et une faibleaccumulation de Ni dans les tissusvgtaux (pour la concentration ne causant aucune inhibition dans la
croissance). Une diminution de
la concentration de mtal dans leau est galement observe. On peutconclure que L. gibba peut tre
un bon candidat pour lpuration des eaux contamines par le cuivre.
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ABSTRACTTo assess the tolerance and phytoaccumulation ability of the duckweed
Lemna gibba L. to copper
(Cu) and nickel (Ni), the aquatic plants were exposed to differentconcentrations of Cu and Ni (0.1
2.0 mg/L) in quarter Coc and Lesaint solution at pH = 6.0 ( 0.1) and
under a daily regime of 12 hlight. Copper and nickel were tolerated by L. gibba at concentrations
0.3 mg/L and 0.5 mg/L,
respectively. However, plant growth decreased by 50% (I50) when themedium contained 0.45 mg/L of
Cu or 0.75 mg/L of Ni. The observed LCI (lowest concentration causingcomplete inhibition) were 0.5
and 1.0 mg/L respectively in the presence of Cu and Ni. Results from
metal analysis in plant tissuesrevealed a high accumulation of copper and a low accumulation of
nickel within the plant (for
concentrations causing no growth inhibition) and a correspondingdecrease of metals in the water.
The duckweed L. gibba L. could be a good candidate for the removal of
low concentrations of copperfrom polluted water.
Certain plants are able to accumulate in their tissues several metals
without showing any signs oftoxicity. This natural accumulation is related with the resistance which
represents response of plants
to metal stress conditions. According to Papazoglou et al. (2005), metalresistance can be based on
either avoidance or tolerance mechanisms. Avoidance reflects the cellprotection against the metal
whereas tolerance is the cell capability to protect themselves against
injury by the metal (Sabreen &sugiyama, 2008). Kanoun-Boul et al. (2008) reported in the tolerance of
duckweed to copper that therelease of organic anions might be involved in the protection of plantsby chelating the metal ions in
the rhizosphere to form non toxic complexes. However, according to
Sabreen and Sugiyama (2008),tolerant populations can be characterized by a lower metal
accumulation than the sensitive one.
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Duckweed commonly refers to a group of floating, flowering plants ofthe family Lemnaceae. The
different species (Lemna, Spirodela, Wolffia and Wolfiella) are
worldwide distributed in freshwater andwetlands, ponds and some effluents are the most common sites to find
duckweed. The plants are fast
growing and adapt easily to various aquatic conditions. They are able togrow across a wide range of
pH, from pH 3.5 to10.5 but survive best between pH 4.5 to 8.3
(Environnement Canada, 1999;Cayuela et al., 2007). The plants are found in temperate climates and
serve as an important foodsource for various water birds and fish (Drost et al., 2007).
Some studies indicate that duckweed plants are sensitive to toxicity.
Other studies however, reportthat duckweed plants are tolerant to environmental toxicity (Wang,
1990). To assess the tolerance of
the species L. gibba to heavy metals, plants were exposed toconcentrations of copper and nickel
higher than those used in medium cultures. Toxic effect of pollutant on
duckweed is generallyevaluated by phytotoxicity tests based on growth inhibition (Geoffroy et
al., 2004). Copper and nickel
were chosen as the metals for this study for a number of reasons. Theirpresence above trace levels
in the environment is an indicator of industrial pollution. On the other
hand, they are essentialmicronutrients for plants; copper is a structural and catalytic
component of many proteins andenzymes involved in metabolic pathways (Teisseire & Vernet, 2000) and
nickel has an important role
in the urease and hydrogenase metabolism (Harish et al., 2008).However, when the concentration
reaches a threshold value, these essential metals become first inhibitoryand afterwards toxic. Copperis responsible for many alterations of the plant cell (respiration,
photosynthesis, pigment synthesis
and enzyme activity) (Teisseire & Vernet, 2000; Kanoun-Boul et al.,2009). Nickel inhibits
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germination, chlorophyll production and proteins (Zhou et al., 2009) inplants; several animal
experimental studies have shown an increased cancer incidence
associated with chronic exposure tonickel.
Each plant species has different resistance and tolerance levels to
different contaminants (Kamal etal., 2004). Therefore, several studies have been performed to elucidate
heavy metal toxicity to plants.
In earlier study, we demonstrated that the duckweed L. gibba L. nativeto the north-east of Algeria
tolerated Zn up to 18 mg/L and was effective in removing 60% of themetal pollutant from the nutrient
medium (Khellaf & Zerdaoui, 2009). The present study investigates
copper and nickel toxicity toduckweed to determine tolerance of this aquatic species to Cu and Ni.
The study also investigates the
potential accumulation of these two metals by the duckweed L. gibba L.The goal was to assess the
possibility to use L. gibba L. for phytoremediation of Cu and Ni
contamination in water.
1.3 . Standards of Disposal
1.4. Treatment methodology
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1.5. Objective and scope of study
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MATERIALS AND METHODS
Preparation steps. Primary treated sewage water were transferred to the
laboratory from the tertiary sewage water treatment plant after the
preliminary sieving step to get rid of large suspended solids. The transferred
water was immediately collected into two opaque tanks (as replicates) to
prevent light entering except at the top (Parr et al., 2002), each tank with
dimensions of 50 cm long, 35 cm wide and 25 cm deep and was filled with
25 L primary treated sewage water. Duckweed (Lemna gibba L.) plants ere
collected from Ganabiet-Tersa drain (Fig. 1). The stock were cleaned by
tap water then washed by distilled water inocula of Lemna plants were
transferred to the water systems for aquatic treatment. The experiment was
kept under outdoor local environmental conditions for eight days retention
time.
Water sampling. Subsurface (under duckweed mt) water samples for
physico-chemical, biological and bacteriological parameters were collected
in polyethylene bottles from all sides of each tank and then mixed. This
procedure carried out every 2 days. Samples volume taken every two days
for each of phytoplankton count and chlorophyll a determination was 100
mL.
Parameters measured. Physico-chemical analyses (Table
I) were carried out according to standard methods for e examination of water
and wastewater (APHA, 1992). Field parameters (pH, conductivity &
dissolved oxygen) were measured in situ using the multi-probe system
(model Hydralab-Surveyor) and rechecked in laboratory using bench-top
equipment to ensure data accuracy for biological parameters including total
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coliform count and fecal coliform. count, phytoplankton identification and
counting andchlorophyll a determination.
Determination of duckweed growth rate. This was determined for fresh
and dry weights. Samples of 20 cm2 area ofLemnaplants were harvested
periodically at the designated time periods (every 2 days) and filtered using
filter papers then fresh weights were determined. These samples were then
dried at 60oC for 48 h to a constant weight and then dry weights were
calculated.
Protien content. Duckweed organic nitrogen content was estimated at the
beginning of the experiment and after 8 days retention time by using Micro-
Kjeldahl method, then the obtained values were multiplied by 6.25 to obtain
protein content values.
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