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INTERNATIONAL JOURNAL OF ENVIRONMENTAL SCIENCES Volume 1, No 7, 2011

Copyright 2010 All rights reserved Integrated Publishing Association

Research article ISSN 0976 4402

Received on April 2011 Published o n June 2011 1702

Phytodegradation of textile dyes by Water Hyacinth (Eichhornia Crassipes)

from aqueous dye solutionsVasanthy Muthunarayanan, Santhiya. M, Swabna. V, Geetha. A

Department of Environmental Biotechnology, School of Environmental Sciences,Bharathidasan University, Trichy-620024, Tamilnadu

[email protected]

ABSTRACT

In this study, the removal of textile dyes, namely Red RB and Black B from their respective

aqueous solutions have been studied using the Water Hyacinth (Eichhornia crassipes). Batch

type experiments were done using the hydrophytes and its dye removal capacity was

observed. The used plant material after the experiment was subjected to GC-MS analysis for

determining the phytochemical components. The remaining waste material was subjected for

composting and the compost produced was characterized in terms of Total Kjeldahl Nitrogen,Total carbon, Total Phosphorus ,pH ,EC and C:N ratio. The above mentioned experiments

have proved the efficiency of Eichhornia crassipes to remove the color and degrade the dyeby about 95% with Red RB and 99.5% with black B. The phytochemical component analysis

indicates the increased production of Hexadecanoic acid, which may be a promising result,but t he reduction in phytol content records a significant reduction in the chlorophyll content.

Keywords:Phytodegradation, Red RB, Black B,Eichhornia crassipes, Phytochemicals

1. Introduction

Synthetic dyes are extensively used in many fields of up to- date technology, e.g., in various

branches of the textile industry (Gupta et al., 1992 Shukla and Gupta, 1992 Sokolowska-Gajda et al., 1996), of the leather tanning industry (Tunay et al., 1999 Kabadasil et al.,

1999) in paper production (Ivanovet al., 1996), in food technology (Bhat and Mathur, 1998

Slampova et al., 2001), in agricultural research (Cook and Linden, 1997 Kross et al., 1996),

in light-harvesting arrays (Wagner and Lindsey, 1996), in photoelectrochemical cells (Wrobel

et al., 2001), and in hair colorings (Scarpi et al., 1998). Unfortunately, the exact amount of

dyes produced in the world is not known. It is estimated to be over 10,000 tons per year.

Exact data on the quantity of dyes discharged in the environment are also not available.

Because of their commercial importance, the impact (Guaratini and Zanoni, 2000) and

toxicity (Walthall and Stark, 1999 Tsuda et al., 2001) of dyes that are released in the

environment have been extensively studied (Hunger, 1995 Calin and Miron, 1995).

Traditional wastewater treatment technologies have proven to be markedly ineffective for

handling wastewater of synthetic textile dyes because of the chemical stability of thesepollutants. A wide range of methods has been developed for the removal of synthetic dyesfrom waters and wastewaters to decrease their impact on the environment. The technologies

involve adsorption on inorganic or organic matrices, decolorization by photocatalysis, and/orby oxidation processes, microbiological or enzymatic decomposition, etc. (Hao et al., 2000).

But for all of these methods phytoremediation proves to be an efficient method.

Phytoremediation is an emerging technology that is rapidly gaining interest and promises

effective and inexpensive cleanup of hazardous waste sites contaminated with metals,


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Phytodegradation of t extile dyes by Water Hyacinth (Eichhornia Crassipes) from aqueous dye solutions

Vasanthy Muthunarayanan, Santhiya. M, Swabna. V, Geetha. AInternational Journal of Environmental Sciences Volume 1 No.7, 2011

1703

hydrocarbons, pesticides, and chlorinated solvents (Macek et al., 2000 Susarla et al., 2002

Xia et al., 2003). The use of plants to degrade, assimilate, metabolize, or detoxify

contaminants is cost-effective and ecologically sound. Four mechanisms are involved in

phytoremediation of organic pollutants: direct uptake and accumulation of contaminants and

subsequent metabolism in plant tissues transpiration of volatile organic hydrocarbons

through the leaves release of exudates that stimulate microbial activity and biochemical

transformations around the root system and enhancement of mineralization at the rootsoilinterface that is attributed to mycorrhizal fungi and microbial consortia associated with the

root surface (Schnoor et al., 1995).The economic success of phytoremediation largelydepends on photosynthetic activity and growth rate of plants.

The water hyacinth Eichhornia crassipes is a floating macrophyte that originated in tropicalSouth America and is now widespread in all tropic climates. It is listed as one of the most

productive plants on earth and is considered one of the world's worst aquatic plants (Epstein,1998). Many large hydropower schemes have to devote significant time and money in

clearing the weed in order to prevent it from entering the turbine and causing damage and

power interruptions.

On the other hand, increased evapotranspiration due to water hyacinth can have seriousimplications where water is already scarce. Water hyacinth can also present many problems

for the fisherman such as decreased fish population, difficult access to the fishing sites and

loss of fishing equipment, resulting in reduction in catch and subsequent loss of livelihood

(Anushree Malik, 2007). Water hyacinth is blamed for the reduction of biodiversity as well.

If it is introduced into foreign aquatic ecosystems, it could cause severe water management

problems because of its vegetative reproduction and high growth rate (Gopal and Sharma,

1981 Giraldo and Garzo n, 2002). However, its enormous biomass production rate, its high

tolerance to pollution, and its heavy-metal and nutrient absorption capacities (Misbahuddin

and Fariduddin, 2002 Trivedy and Pattanshetty, 2002 Williams, 2002 Singhal and Rai,

2003 Ghabbour et al., 2004 Jayaweera and Kasturiarachchi, 2004) qualify it for use inwastewater treatment ponds.

Water hyacinth (Eichhornia crassipes Solms), due to its fast growth and large biogasproduction (Singhal and Rai, 2003), has potential to cleanup various wastewaters. Inorganic

contaminants such as nitrate, ammonium and soluble phosphorus (Reddyet al., 1982 Reddy,1983), heavy metals (Muramoto and Oki, 1983 Zhu et al., 1999) can be removed efficiently

by water hyacinth through uptake and accumulation. Previously the roots of water hyacinthplants and their roots were used for phytoremediation of ethion and biosorption of reactive

dyes (Huilong Xia, Xiangjuan Ma, 2005). The objective of this study is to use Eichhornia for

dye removal and to subject the plant further for composting.

2. Materials and Methods

2.1 Dyes used for the study

The reactive dyes used as adsorbates for the study were Red RB and Black B. The structures

of these dyes are elucidated below:


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Figure 1:Structure of Reactive Red 198 Figure 2:Structure of Reactive Black 5

2.2 Dye solutions

Dye stock dye solutions of both the dyes were prepared by dissolving 100 mg of dye in 100ml sterile distilled water to get 1000 ppm dye solution. A suitable aliquot of the sample

solution containing dye was transferred into a 100 ml volumetric flask and the solution was

made up to the mark with double distilled water. The absorbance was measured at the

respectivelmax against a blank. A standard graph was plotted for 10 100 mg/L of dye.

2.3 Preparation of live plants

Plants of E. crassipes were obtained from local pond near Ariyalur city and from TANCEM

mines near Ariyalur city respectively. The plants were washed thoroughly. The fresh plantswere grown under laboratory conditions. The plants were used for the present investigation.

The plants were grown in a nutrient solution which was renewed once a week. The nutrientsolution contained N (NH4NO3), 38 mgl_1 P (KH2PO4), 3.5 mgl_1 K (KCl), 30 mgl_1 Ca

(CaCl2 2H2O), 9 mgl_1 Mg (MgSO4 7H2O), 7 mgl_1 trace elements such as Fe, Mn,B, Zn, Mo, Cu, and Co at the concentrations of 3, 0.45, 0.12, 0.16, 0.05, 0.005, and 0.005

mgl_1, respectively (Huilong Xia, Xiangjuan Ma, 2005).

2.4 Outdoor cascade experiments (set nos. 1 and 2)

The first and second sets of laboratory experiments were both performed with five identicalcontainers (0.39 - 0.56m

2and 0.45 - 0.7m

2floor area, respectively). These containers were

operated at 10 L levels of aqueous dye solutions of concentrations ranging from 10, 20, 30,

40 and 50 ppm (each) were prepared for both the sets.

2.4.1 Set no 1

In five containers of Red RB aqueous dye solutions, which were set as a cascade, floating Eichhornia crassipes plants (12 pieces in each) were introduced and one container each

(without plants) for all the five concentrations of aqueous dye solutions was maintained as the

controls (Table 1).The color reduction in the cascades were checked in terms of optical

density in all the concentrations at different time intervals namely 24,48,72,96,120 and 144

hours. The max for the dye was 519 nm.


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2.4.2 Set no 2

The similar set up was also used for Black B aqueous dye solutions. In five containers of

Black B aqueous dye solutions, which were set as a cascade, floating Eichhornia crassipes

plants (12 pieces in each) were introduced and one container each (without plants) for all the

five concentrations of aqueous dye solutions was maintained as the controls (Table 1). The

color reductions in the cascades were checked in terms of optical density in all theconcentrations at different time intervals namely 24,48,72,96,120 and 144 hours. The maxfor the dye was 597 nm.

Table 1:Outdoors pulsed cascade setup

AqueousReactive

dyes

Concentrationof dye (ppm)

Biomass ofEichhornia crassipes

(gm)

Time (hrs)

Red RB 10-50 400

Black B 10-50 400

24

48

72

96120

144

168

2.5 Determination of Phytochemical components present in the plant material.

(Eichhornia crassipes)(Kokate, 1993)

About 4 gm of powdered plant material was soaked in 20 ml of absolute alcohol overnightand then filtered through whatmann filter paper No.41 along with 1gm sodium sulfate to

remove the sediments and traces of water in the filtrate. Before filtering, the filter paper alongwith sodium sulphate was wetted with absolute alcohol. The filtrate was then concentrated by

bubbling nitrogen gas into the solution and the volume was reduced to 1ml. This extract wasanalyzed using GS-MS for the analysis of phytochemical components of the plant materials

used.

2.6 Preparation of compost (Reuse of the used filter papers and plant materials)

The waste plant materials (Eichhornia sp.,) obtained after the pilot scale treatment of the

aqueous dye solutions and used filter papers were subjected for the process of composting

along with garden waste and cow dung (Table 2). The pre composted compost was further

subjected to vermicomposting using the earthworm speciesPerionyx excavatus.

Table 2:Preparation of Compost

S.No. Component Weight (Kg) Total Weight (Kg)

1

Cow dung + Garden

waste + Eichhornia

used plants + used filter

papers

2 kg + 1 kg +

3 kg6


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2.7 Analysis of the compost

After 60 days the compost was taken from the pit and was subjected for the analysis. The pH,

EC, available Nitrogen, Potassium, Phosphorus, C/N ratio were estimated. (Muthuvel and

Udayasoorian, 1999).

3. Test results and discussion

3.1 Color reduction in Outdoor Cascade Experiments

The maximum color reduction was observed at 144 hours after the introduction of thefloating and submerged plants into the 10 ppm RB and Black B aqueous dye solutions. It

accounts for 95% removal in Red RB dye and 99.5 % in Black B dye in 10 ppm aqueous dyesolution at 144 hours respectively. In the 50 ppm aqueous dye solutions the color removal

was observed after 6 days at the rate of 71.7% removal in Red RB dye and 76.7%respectively (Tables 3 and 4 ) (Figures 1 and 2) . Similarly Vasanthy et al., (2006) has

checked the treatability of aqueous Majanta HB solutions (5, 10, 15, 20 and 25 ppm) usingEichhornia crassipes. The plant saplings were found to remove 95% color from 50ppm dye

solution after 6 days. The highest color removal obtained from 25 ppm dye solution was70% after 144 hours.

Table 3:Effect of time on percent dye (Red RB) removal usingEichhornia crassipes

Percentage (%)Concentration

24 hrs 48 hrs 72 hrs 96 hrs 120 hrs 144 hrs

10

20

30

40

50

65.4

56.9

38.4

27.3

16.9

69.1

63.1

55.2

42.7

39.7

76.4

69.6

64.7

54.7

54.1

79.5

71.5

66.2

62.7

59.8

83.5

78.5

77.2

72.6

67.9

95

87.7

83.1

80

71.7

The removal of the aqueous dyes may be due to Biosorption i.e., the sorption of dye

molecules onto the root, shoot and the leaves of the plant. Similar result has been put forth by

Vengata Mohan et al., 2002. Interestingly, the insight into the speciation and localization of

dyes in plant tissues also provides a due rate and extent of uptake by particular plant parts. Itis often observed that roots accumulate much higher concentration of pollutants (Anushree

Malik, 2007).The efficiency may be due to the fact that the biological processes has the

potential to convert or degrade the pollutants into water,CO2 and various salts of inorganic innature. The complete breakdown of an organic molecule into inorganic component should bethe desired outcome to avoid the persistence of potentially hazardous components in the

environment.


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Table 5:Phytochemical components identified in the water hyacinth (Eichhornia crassipes)

before treatment

No RT Name of the

compound

Molecular

Formula

MW Peak

Area

Compoun

d Nature

Activity*/*

1 2.92 Butane, 1,1-

diethoxy-C8H18O2 146 6.60 Et her No activity reported

2 5.91 Propane, 1,1,3-

triethoxy-C9H20O3 176

6.50

Ether No activity reported

3 7.54 4H-Pyran-4-one,

2,3-dihydro-3,5-

dihydroxy-6-methyl-

C6H8O4 144

0.65

Flavonoid

fraction

No activity reported

4 8.75 Methyl Salicylate C8H8O3 152

0.80

Analgesic

compound

Antipyretic, Anti-

inflammatory,

Analgesic, Antiseptic,

Pesticide, Insectifuge,Cancer- preventive,

Carminative, Perfumery

5 11.5

8

Pipradrol C18H21N

O

267

0.35

Alkaloid Antimicrobial

6 12.7

7

1-(2,4-

dihydroxyphenyl)-2-(4-methoxy-3-

nitrophenyl)ethanon

e

C15H13N

O6

303

0.18

Phenolic

compound

Antimicrobial

7 13.2

8

Nonanoic acid, ethyl

esterC11H22O

2

186

0.41

Fatty acid

ester

Antimicrobial

8 14.0

9

N-Phenethyl-2-

methylbutylidenimine

C13H19N 189

0.31

Nitrogen

compound


9 16.9

8

1H-Pyrrole, 1-

phenyl-C10H9N 143

0.14

Alkaloid Antimicrobial

10 17.4

0

Nonanoic acid C9H18O2 158

0.43

Fatty acid Antimicrobial

11 17.7

4

1-Amino-2-

methylnaphthaleneC11H11N 157

0.18

Aromatic

compound

Insecticide

12 17.8

1

Diethyl Phthalate C12H14O

4

222

0.53

Plasticizer

compound


13 23.5

7

3,7,11,15-

Tetramethyl-2-

hexadecen-1-ol

C20H40O 296

44.4

6

Terpene

alcohol

Antimicrobial

14 23.7

5

Didodecyl phthalate C32H54O

4

502

7.62

Plasticizer

compound


15 24.4

3

3,7,11,15-

Tetramethyl-2-

hexadecen-1-ol

C20H40O 296

1.29

Terpene

alcohol

Antimicrobial

16 25.8

9

n-Hexadecanoic

acidC16H32O

2

256

14.4

0

Palmitic

acidAntioxidant, Hypocholesterolemic

Nematicide, Pesticide, Lubricant,

Antiandrogenic, Flavor, Hemolytic 5-

Alpha reductase inhibitor

17 28.8

3

Phytol C20H40O 296 21.1

2

Diterpene Diuretic, Antimicrobial,

Anticancer


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Table 6:Phytochemical components identified in the water hyacinth (Eichhornia crassipes)

after treatment

No RT Name of the

compound

Molecular

Formula

M

W

Peak

Area

Compound

Nature

Activity**

1 2.92 Butane, 1,1-

diethoxy-

C8H18O2 146 0.91 Ether No activity reported

2 5.92 Propane, 1,1,3-

triethoxy-C9H20O3 176 8.50 Ether No activity reported

3 7.55 4H-Pyran-4-one,

2,3-dihydro-3,5-

dihydroxy-6-

methyl-

C6H8O4 144 0.68 Flavonoid

fraction


4 8.51 Octanoic acid,

ethyl ester

C10H20O

2

172 0.19 Fatty acid ester Insecticide

5 8.76 Methyl

SalicylateC8H8O3 152 1.05 Analgesic

compound

Antipyretic, Anti-

inflammatory,

Analgesic,

Antiseptic, Pesticide,Insectifuge,

Cancer- preventive,

Carminative,

Perfumery

6 12.78 2',4'-

Dihydroxypropi

ophenone

C9H10O3 166 0.28 Phenolic

compound

Antimicrobial

7 13.29 Decanoic acid,

ethyl esterC12H24O

2

200 0.63 Fatty acid ester Antimicrobial

8 17.40 Nonanoic acid C9H18O2 158 1.42 Antimicrobial

9 17.81 Diethyl

PhthalateC12H14O

4

222 0.55 Plasticizer

compound


10 23.56 3,7,11,15-

Tetramethyl-2-

hexadecen-1-ol

C20H40O 296 37.84 Terpene

alcohol

Antimicrobial

11 23.73 Didodecyl

phthalate

C32H54O

4

502 8.22 Plasticizer

compound


12 24.42 3,7,11,15-

Tetramethyl-2-

hexadecen-1-ol

C20H40O 296 11.29 Terpene

alcohol

Antimicrobial

13 25.86 n-Hexadecanoicacid

C16H32O2

256 17.79 Palmitic acid Antioxidant, Hypocholesterolemic

Nematicide,

Pesticide, Lubricant,

Antiandrogenic, Flavor,

Hemolytic, 5-Alpha reductase

inhibitor

14 28.79 Phytol C20H40O 296 10.67 Diterpene Diuretic AntimicrobialAnticancer

Anti-inflammatory


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3.2 Analysis of the phytochemical components through GC-MS

The phytochemical components of Eichhornia crassipes have shown changes before and

after treatment (Tables 5 and 6) (Figures 3 & 4).The aromatic components of plant

determined includes Butane, 1,1 diethyl, Propane, Nonanoic acid, n-Hexadecanoic acid and

Phytol. Butane and Propane compounds have been reported both before and after treatment

and the peak area have increased from 6.50% to 8.50% for n-Hexadecanoic acid. The peakarea for phytol has got decreased from 21.12 to 10.67 (Table 6). However Chlorophyll is themost abundant photosynthetic pigment in higher plants. Normally, chlorophyll is reported to

be hydrolyzed, resulting in the release of free phytol and chlorophyllide. Although thedegradation of chlorophyllide has been studied in depth, the metabolic fate of phytol in plants

is reported to be less clear. But the reduction in phytol content may be interpreted due to thereduction in chlorophyll which may be due to the stress posed by the dye stuffs. Further

Puvaneswari et al (2006) reported that industrial effluents could increase the enzymechlorophyllase, which is responsible for the chlorophyll degradation or decrease in the

cytokinins which stimulates chlorophyll synthesis.

Production of n-Hexadecanoic acid during the degradation of textile dyes has been reported

earlier. The compound is found to be a biopolymer which can be degraded rapidly between125 C to 225 C (Dhawal P. Tamboli et al, 2010). Hence the polymer that is produced

during dye degradation can be purified and used for further biopolymer studies after further

investigation.

PPRCTANJORE, 24-Sep-2007 + 11:13:3Water Hyacinth alcohol ext-Black dye

6.67 11.67 16.67 21.67 26.67 31.67 36.67Time0

100

%

Medicinal plant analysis164 Scan EI+

TIC6.49e7

23.55

5.92

2.92 5.39 20.3317.408.75

25.8623.73 38.6435.89

28.7829.59

Figure 3:Phytochemical components of water hyacinth before treatment (Control)

3.3 Compost Analysis

The treated plants were then subjected to composting along with used filter papers, cow dung

and neem leaves. The various components such as total nitrogen, total phosphorus, total

potassium, organic carbon, organic matter and C: N ratios were analyzed. The amount of

nitrogen before composting was found to be 1.47% and it has increased to 2.51% after the

process.


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Figure 4:Phytochemical components of water hyacinth after treatment (Affected)

3.3 Compost Analysis

The treated plants were then subjected to composting along with used filter papers, cow dung

and neem leaves. The various components such as total nitrogen, total phosphorus, total

potassium, organic carbon, organic matter and C: N ratios were analyzed. The amount ofnitrogen before composting was found to be 1.47% and it has increased to 2.51% after theprocess. The total organic carbon content has reduced from 42.08% to 25.45%. The

potassium and phosphorus content have been recorded to be 0.62% and 0.49% respectivelyin the initial mass and after composting it has changed to 0.98% and 0.75%. The carbon

nitrogen ratio has got reduced from 28.63:1 to 10:1 (Table 7).

Table 7:Manural value of the compost

ValuesS.No. Parameters

Initial Final

1.

2.3.

4.

5.

6.

Organic Carbon %

Total Nitrogen %Total Phosphorus%

Total Potassium %

C:N

Colour

42.08

1.470.49

0.62

28.63:1

Yellowish brown

25.45

2.510.75

0.98

10:1

Brown

The initial organic carbon has varied from 42 to 25.45%. The TOC has decreased as the

decomposition progressed. Similarly, the organic carbon was recorded to get reduced for the

vegetable and fruit waste subjected for Vermicomposting (by about 83%). This could be

attributed to the faster decomposition of carbon present in the form of lignin in vegetable and

fruit waste by earthworms (Susila, 2009). Similarly Goyal et al. (2005) reported the lowest

organic carbon in the water hyacinth waste and Atkinson et al. (1996) reported that during

poultry waste decomposition with sawdust, about 29% of carbon reduction had occurred. Asimilar result has been obtained by Garcia et al. (1991). Normally, in all the composting

mixtures the carbon content has been found to be reduced and the nitrogen content increased,

thus causing a general decrease in the C: N values. The C: N ratio helps to gauge how far the

process has gone (Troeh and Thompson, 2005). Pandey (2009) have reported a C: N ratio of8.15 in poultry manure amended compost. In the mixtures the range of C: N ratio was about

79. Many such wastes have been found to be readily decomposable by soil microbes. Thus,the decomposition of organic matter reduces the amount of TOM and leaves the compost

enriched with nitrogen. The C: N ratio has reduced substantially from 28.63:1 to 10:1 .A


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decline of C: N ratio to less than 20 indicate an advanced degree of organic matter

stabilization and reflects a satisfactory degree of maturity of organic wastes according to

Senesi (1989). As the decomposition progressed during the composting process, the carbon

content has reduced as it was lost as carbon dioxide and the N content has increased as the

complex proteinaceous material has broken down into simpler N containing compounds like

ammonia. This metabolic trend has ultimately reduced the C: N of Eichhornia wastes

subjected for Vermicomposting. Li et al. (2001) have recorded the reduction in C: N ratioduring composting process and inferred that the reduction in carbon and lowering of C: N

ratio in the Vermicomposting process could be achieved either by the respiratory activity ofearthworms and microorganisms or by increase in nitrogen by microbial mineralization of

organic matter in combination with the addition of the worms nitrogenous waste throughtheir excretion (Christry and Ramalingam, 2005).

The initial and final TKN of the waste subjected for vermicompost were 1.47 and 2.51respectively. As the C: N ratio of all the wastes were relatively higher initially, there was not

much loss of N as ammonia and hence the N content increased with days. (Goyal et al.,

2005 SanchezMonedero et al., 2001 Reddy et al. 1979). Hence the present study has

established the fact that the used plant materials along with the cow dung and leaf wastes

could be very well subjected for the process of vermicomposting. And it has resulted in acompost material with a favorable C:N ratio. Similarly Umamaheswari et al., (2006) have

checked the possibility of converting Eichhornia as compost. Further the compost has been

used for the germination ofAbelmoschus esculentus.

4. Conclusion

As per the study the promising attributes of Water hyacinth includes its tolerance to dye and

dye absorption along with good root development, low maintenance and ready availability in

contaminated regions. These characteristics prove the suitability of water hyacinth in dyeing

industry effluent treatment ponds. However further experiment could be done to optimize the

conditions for the treatment of the direct effluents and caution must be always taken as these

Hydrophytes can easily contaminate the aquatic ecosystem.

1. The above mentioned experiment has proved the efficiency ofEichhornia crassipestoremove the color and degrade the dye by about 95% with Red RB and 99.5% with

black B.

2. The phytochemical component analysis indicates the increased production of

Hexadecanoic acid, which may be a promising result, but the reduction in phytolcontent records a significant reduction in the chlorophyll content which needs further

investigation.

3. It has been further established by subjecting the Eicchornia plants used for treatment

for vermicomposting with cow dung and leaves.

4. The process further solves the solid waste disposal problem also and could be

accepted as a reliable method for dye degradation and solid waste reduction.

5. References

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