Phytodegradation of textile dyes by Water Hyacinth ... · The similar set up was also used for...

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 on June 2011 1702

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

Vasanthy Muthunarayanan, Santhiya. M, Swabna. V, Geetha. A Department of Environmental Biotechnology, School of Environmental Sciences,

Bharathidasan University, Trichy620024, 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 GCMS 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 dye by 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 the 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 (Tu¨nay et al., 1999; Kabadasil et al., 1999) in paper production (Ivanov et 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 lightharvesting 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 these pollutants. A wide range of methods has been developed for the removal of synthetic dyes from waters and wastewaters to decrease their impact on the environment. The technologies involve adsorption on inorganic or organic matrices, decolorization by photocatalysis, and/or by 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,


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

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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 costeffective 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 root–soil interface that is attributed to mycorrhizal fungi and microbial consortia associated with the root surface (Schnoor et al., 1995).The economic success of phytoremediation largely depends on photosynthetic activity and growth rate of plants.

The water hyacinth Eichhornia crassipes is a floating macrophyte that originated in tropical South 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 serious implications 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, it’s enormous biomass production rate, its high tolerance to pollution, and its heavymetal 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 in wastewater treatment ponds.

Water hyacinth (Eichhornia crassipes Solms), due to its fast growth and large biogas production (Singhal and Rai, 2003), has potential to cleanup various wastewaters. Inorganic contaminants such as nitrate, ammonium and soluble phosphorus (Reddy et 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 hyacinth plants 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 100 ml 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 respective λ max 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 plants were 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 nutrient solution contained N (NH4NO3), 38 mgl_1; P (KH2PO4), 3.5 mgl_1; K (KCl), 30 mgl_1; Ca (CaCl2 Æ 2H2O), 9 mgl_1; Mg (MgSO4 Æ 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 identical containers (0.39 0.56m 2 and 0.45 0.7m 2 floor 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 λ 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 the concentrations at different time intervals namely 24,48,72,96,120 and 144 hours. The λ max for the dye was 597 nm.

Table 1: Outdoors pulsed cascade setup

Aqueous Reactive dyes

Concentration of dye (ppm)

Biomass of Eichhornia crassipes (gm)

Time (hrs)

Red RB 1050 400

Black B 1050 400

24487296 120 144 168

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 overnight and 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 along with 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 was analyzed using GSMS 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 species Perionyx 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 kg 6



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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 the floating 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 dye solution 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) using Eichhornia 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 was 70% after 144 hours.

Table 3: Effect of time on percent dye (Red RB) removal using Eichhornia 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. It is 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 in nature. The complete breakdown of an organic molecule into inorganic component should be the desired outcome to avoid the persistence of potentially hazardous components in the environment.



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Table 4: Effect of time on percent dye (Black B) removal using Eichhornia crassipes

Percentage (%) Concentration 24 hrs 48 hrs 72 hrs 96 hrs 120 hrs 144 hrs

10

20

30

40

50

74.25

68.69

62.28

58.63

51.94

79.48

74.31

67.12

62.37

58.97

86.4

76.46

71.51

68.84

66.11

89.5

81.75

75.62

72.87

69.18

93.5

88.29

79.2

74.6

67.9

99.5

94.77

83.1

80

76.7

0

10

20

30

40

50

60

70

80

90

100

24 48 72 96 120 144

Time (hours)

10 ppm 20 ppm 30 ppm 40 ppm 50 ppm

Figure 1: Effect of time on percent dye (Red RB) removal using Eichhornia crassipes

Figure 2: Effect of time on percent dye (Black B) removal using Eichhornia crassipes

0

20

40

60

80

100

120

24 48 72 96 120 144

Time (hours)

Percentage removal

10 ppm

20 ppm

30 ppm

40 ppm

50 ppm



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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 Ether No activity reported

2 5.91 Propane, 1,1,3 triethoxy

C9H20O3 176 6.50

Ether No activity reported

3 7.54 4HPyran4one, 2,3dihydro3,5 dihydroxy6methyl

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 (4methoxy3 nitrophenyl)ethanon e

C15H13N O6

303

0.18

Phenolic compound

Antimicrobial

7 13.2 8

Nonanoic acid, ethyl ester

C11H22O 2

186 0.41

Fatty acid ester

Antimicrobial

8 14.0 9

NPhenethyl2 methylbutylidenimin e

C13H19N 189

0.31

Nitrogen compound


9 16.9 8

1HPyrrole, 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

1Amino2 methylnaphthalene

C11H11N 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 Tetramethyl2 hexadecen1ol

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 Tetramethyl2 hexadecen1ol

C20H40O 296

1.29

Terpene alcohol

Antimicrobial

16 25.8 9

nHexadecanoic acid

C16H32O 2

256

14.4 0

Palmitic acid

Antioxidant, 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

MW

Peak Area

Compound Nature

Activity**

1 2.92 Butane, 1,1 diethoxy


2 5.92 Propane, 1,1,3 triethoxy


3 7.55 4HPyran4one, 2,3dihydro3,5 dihydroxy6 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 Salicylate

C8H8O3 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 ester

C12H24O 2

200 0.63 Fatty acid ester Antimicrobial

8 17.40 Nonanoic acid C9H18O2 158 1.42 Antimicrobial

9 17.81 Diethyl Phthalate

C12H14O 4

222 0.55 Plasticizer compound


10 23.56 3,7,11,15 Tetramethyl2 hexadecen1ol

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 Tetramethyl2 hexadecen1ol

C20H40O 296 11.29 Terpene alcohol

Antimicrobial

13 25.86 nHexadecanoic acid

C16H32O 2

256 17.79 Palmitic acid Antioxidant, Hypocholesterolemic

Nematicide,

Pesticide, Lubricant,

Antiandrogenic, Flavor,

Hemolytic, 5Alpha reductase

inhibitor 14 28.79 Phytol C20H40O 296 10.67 Diterpene Diuretic Antimicrobial

Anticancer Antiinflammatory



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3.2 Analysis of the phytochemical components through GCMS

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, nHexadecanoic 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 nHexadecanoic acid. The peak area for phytol has got decreased from 21.12 to 10.67 (Table 6). However Chlorophyll is the most abundant photosynthetic pigment in higher plants. Normally, chlorophyll is reported to be hydrolyzed, resulting in the release of free phytol and chlorophyllide. Although the degradation 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 the reduction 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 enzyme chlorophyllase, which is responsible for the chlorophyll degradation or decrease in the cytokinins which stimulates chlorophyll synthesis.

Production of nHexadecanoic acid during the degradation of textile dyes has been reported earlier. The compound is found to be a biopolymer which can be degraded rapidly between 125° 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.

PPRC TANJORE, 24Sep2007 + 11:13:34 Water Hyacinth alcohol extBlack dye

6.67 11.67 16.67 21.67 26.67 31.67 36.67 Time 0

100

%

Medicinal plant analysis164 Scan EI+TIC

6.49e7 23.55

5.92

2.92 5.39 20.33 17.40 8.75

25.86 23.73 38.64 35.89

28.78 29.59

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 of nitrogen before composting was found to be 1.47% and it has increased to 2.51% after the process. 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% respectively in 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

Values S.No. Parameters Initial Final 1. 2. 3. 4. 5. 6.

Organic Carbon % Total Nitrogen % Total Phosphorus% Total Potassium % C:N Colour

42.08 1.47 0.49 0.62 28.63:1 Yellowish brown

25.45 2.51 0.75 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. A similar 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 of 8.15 in poultry manure amended compost. In the mixtures the range of C: N ratio was about 7–9. 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 ratio during 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 of earthworms and microorganisms or by increase in nitrogen by microbial mineralization of organic matter in combination with the addition of the worm’s nitrogenous waste through their excretion (Christry and Ramalingam, 2005).

The initial and final TKN of the waste subjected for vermicompost were 1.47 and 2.51 respectively. 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 ; Sanchez–Monedero 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 a compost 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 of Abelmoschus 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 of Eichhornia crassipes to remove 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 phytol content 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.

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