Phytoremediation of Metals Contaminated Distillery Effluent Using Water Hyacinth (Eichhornia crassipes)

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1 International Journal of Engineering Technology, Management and Applied Sciences Phytoremediation of Metals Contaminated Distillery Effluent Using Water Hyacinth (Eichhornia crassipes) Dr. Poonam Bathla Department of Applied Science, First year Engineering Vidyalankar Institute of Technology, Mumbai-37 Abstract: Phytoaccumulation potential of water hyacinth (E.crassipes) against anaerobically treated distillery effluent was studied.metal content (Fe, Zn, Na, K, Mg and Ca) in the effluent reduced with increasing time period after treatment with water hyacinth (E. crassipes). Maximum reduction was observed at 2% effluent concentration after 15 days. Metal content in water hyacinth plant increased with increasing time period. Maximum increase was observed after 15 days of treatment at 8% effluent concentration. Total chlorophyll and free proline content in water hyacinth was studied to analyze stress due to metal uptake. Total Chlorophyll in water hyacinth plant increased at 2% effluent concentration, and decreased thereafter.maximum decrease was observed at 8 % effluent concentration after 15 days of treatment. Free proline content in water hyacinth was not affected at 2 % effluent concentration; however it increased with the further increase in effluent concentration. Maximum increase was observed at 8% effluent concentration after 15 days of treatment. Key words: Distillery effluent, water hyacinth, phytoremediation, heavy metals, spent wash Introduction : Rapid industrial development and population explosion has created problems of environmental pollution which are gaining severe dimensions day by day all over the world, in general, and in developing countries in particular. Water pollution by industrial wastes has become amajor problem in India. Distilleries are one of the major polluting industries with 88% raw material ending up as waste. The large volume of effluent generated during the production of alcohol from the distillation section, contributes to major fraction of waste water from distilleries. Distillery effluent also known as spentwash is one of the most complex, caramelized, and cumbersome waste having very high BOD (35,-5, ppm), COD (85,-1, 3, ppm) and high concentration of heavy metals. The disposal of distillery effluent results in massive destruction of aquatic flora and fauna, deterioration of water quality of the receiving streams and possess serious public health hazards. Phytoremediation is novel eco-friendly and cost effective approach used for treatment of waste water. Phytoremediation describes the use of plants, taking advantage of their ability to concentrate elements and compounds from the environment and to metabolize various molecules in their tissues.several aquatic plants have been employed for the removal of organic and inorganic matter from waste water, among them aquatic free floating macrophytes are found to be very effective as they are easily available and have capability to biologically concentrate heavy metals and synthetic organic chemicals. This paper reviews phytoaccumulation efficiency of water hyacinth (Eichhornia crassipes) as a cost effective approach for removal of metals from distillery wastewater. Materials and Methods : The distillery effluent was collected from Kesar Enterprises, Baheri (U.P.), before and after primary (anaerobic) treatment.the effluent was analyzed for metal content and other parameters as per standard methods (APHA,1995).The water hyacinth plant viz Eichhornia crassipes was collected from a natural pond on road side, Lalkuan. After collection, the plants were washed thoroughly with running tap water followed by distilled water to avoid surface contamination and were kept in pots containing water for 24 hours prior to experimentation for acclimatization. To access the phytoaccumulation efficiency of water hyacinth against distillery effluent, four concentrations (2, 4, 6 and 8%) were prepared with water. The effluent concentrations for phytoremediation efficiency evaluation were decided on basis of preliminary experiments, which showed almost no growth of these plants beyond % effluent concentration.for experiment, five sets of triplicate pots were filled with 2 litre of distillery effluent of different concentration (2, 4, 6, and 8%). Two plants of average size were allowed to grow in 283 Dr. Poonam Bathla

2 International Journal of Engineering Technology, Management and Applied Sciences each pot for 15 days. One set of triplicate pots, filled with % effluent (water) and plant was maintained as control. In order to provide oxygen to plant roots mechanical aeration was given by air pump at rate of min/pot at 2 days interval throughout the experiment.effluent samples from all the pots were then analyzed after 5, and 15 days for heavy metal with the help of Atomic Absorption Spectrometer (AAS) using nitric acid-perchloric acid digestion (APHA, 1995). Simultaneously, the plant samples were analyzed for total chlorophyll, free proline content and heavy metals (Arnon, 1949; Bates et al. 1973; APHA 1995). Results anddiscussions : A. Physico-chemical Characteristics of Effluent Physico-chemical characteristics of the raw and anaerobically treated effluent were analyzed and the data are given in Table1. In general, the results of the present study are in agreement with other studies on distillery effluent (Chandra and Singh, 1999; Chanrda and Pandey, 2; Vaidyanathan et al., 1995; Singh and Kumar, 1995). Discharge parameters are much higher than BIS standards (Bhasin et al., 27) and therefore requires treatment before being disposed in nearby water streams. Table 1: Physicochemical characteristics of Raw and Anaerobically treated distillery effluent S.No. Parameter Raw** Anaerobically Treated* 1. ph 4.11± ± Colour Dark brown Greenish brown 3. Odour Alcoholic Sugary 4. Colour (cu) ± ± COD (ppm) 86± ± Total solids (ppm) ± ± Total Nitrogen (ppm) ± ± Total Phosphorous (ppm) 769.5± ± Cobalt (ppm).87±.1.355±.12. Cadmium (ppm).656± ± Iron(ppm) 49.43± ± Zinc(ppm) 4.495± ± Sodium(ppm) ± ± Potassium(ppm) 39.3± ± Magnesium(ppm) 17.8± ± Calcium(ppm) ± ±27.18 * Each reading is the mean of eight samples ± SE ** Each reading is the mean of three samples with ± SE B.Phytoaccumulation of Metals from Distillery Effluent Water hyacinth has been employed for phytoremediation of metals (Fe, Zn, Na, K, Mg and Ca) from the distillery effluent. At all the effluent concentrations reduction in various metals was observed with increasing time periods. 284 Dr. Poonam Bathla

3 Reduction (%) Reduction (%) International Journal of Engineering Technology, Management and Applied Sciences The reduction in metals at various effluent concentrations is associated with uptake of these metals through plant roots from the effluent. At 2% effluent concentration, the reduction was maximum (67.7, 55.55, 49.61, 52.9, and 45.8%) for Fe, Zn, Na, K, Mg and Ca, respectively after 15 days. Further reduction in metal content decreased with increasing effluent concentration and was minimum at 8% effluent concentration after 15 days ( 57.48, 42.85, 41.92, 4.27, and 33.47%) for iron, zinc, sodium, potassium, calcium and magnesium respectively as presented in Figure 1and 2. Removal of metals is likely to be due toplant uptake. As most of heavy metals are positive ions, they are attracted towards the negative charges born on roots of water hyacinth while decrease in reduction at higher concentration may be due to metal toxicity induced at higher concentration (Basu et al., 23).In the past, various macrophytes and microphytes have been employed for removal of various metals from the effluents (Abbasi and Ramasami, 1999; Gopal et al., 1991; Ackin et al., 1993 and Zhu et al., 1999). Srivastav and Pandey (1999) reported decrease in Fe, Zn, Cu and Mn content using E. crassipes and Spirodela polyrrhiza against paper mill effluent. Aquatic macrophytes have capability of bio-concentrating heavy metals and synthetic organic compounds as reported by O Breen (1981) and Sulton and Kenenth (1983). Singhal et al.(23) and Upadhyay (23) reported metal accumulation by water hyacinth when employed for treatment of pulp and paper, distillery and brass industry effluent. Selvapathy et al.(1997) also reported metal accumulation from waste water using water hyacinth. Gupta and Sujatha (1996) also reported metal accumulation from tannery wastewater using water hyacinth. Water hyacinth has been found to be an effective agent for reduction of metals present in distillery effluent, because of its high efficiency of removing pollutants from industrial waste water, especially organic pollutants and heavy metals which is in agreement with the results of present study Effluent concentration (2%) 5 15 Fe Zn Na K Ca Mg Fig. 1: Reduction in metal content of distillery effluent ( 2% ) after treatment with water hyacinth (E. crassipes) 6 Effluent concentration (8%) 5 Fe Zn Na K Ca Mg Fig. 2: Reduction in metal content of distillery effluent (8%) after treatment with water hyacinth (E. crassipes) 285 Dr. Poonam Bathla

4 Reduction (%) International Journal of Engineering Technology, Management and Applied Sciences B. Changes in Chlorophyll, Free Proline, and Metal Content of Water Hyacinth a. Changes in Chlorophyll Content of Water Hyacinth The present study shows that chlorophyll a,chlorophyll b and total chlorophyll (a+b) content was even greater at 2% effluent concentration over the control (water), % effluent concentration (Fig 3). Except at 2% effluent concentration; chlorophyll content declined further with increasing effluent concentration. At 2% effluent concentration the chlorophyll content increased with increasing time period upto days, and after that no further increase was observed. Similarly Chlorophyll content decreased at 4, 6 and 8 % effluent concentration up to days and after that there was no change in the total chlorophyll content. Similar results were reported by Srivastav and Pandey (1999) when E. crassipes, P. stratiotes and H. verticillataon were exposed to different concentrations of effluent for different durations. Minimum reduction in total chlorophyll was observed at lower concentration and it increased with increasing treatment duration. Basu et al. (23) reported chlorophyll content did not show significant change at lower concentration. However, at higher concentration 98% loss in chlorophyll was observed. Similar type of results were reported by Singhal et al. (23) and Upadhyay (23) when channel grass and water hyacinth were employed for phytoremediation of pulp and paper, distillery and brass industry effluent. The reduction in chlorophyll was associated with the uptake of toxicant by the plant from effluent. Izawa (1979) suggested that the inhibition of chlorophyll may be possible due to the effluent induced inhibition of electron transport system in photosystem-ii, the fall in chlorophyll content under higher concentration might be due to inhibitory effect of toxicant. Wong and Chang (1991) also associated inhibition of chlorophyll synthesis in plant with toxicant in exposure medium % 2% EC 4% EC 6% EC 8% EC Fig 3: Reduction in total chlorophyll (a+b) content of water hyacinth (E. crassipes) during phytoremediation of distillery effluent b. Changes in Free Proline Content of Water Hyacinth Effect of different concentration of distillery effluent on free proline content of E. crassipes at varying time intervals are shown in Fig 4.Free proline content increased with increasing time period at 4, 6, and 8% effluent concentrations, however the rate of increase in proline content slowed down after days. At % (water) and 2% effluent concentration, the proline content was almost the same with increasing the period. At 8% effluent concentration it was 5.7, and 89.9% after 5,, and 15 days, respectively. Except at 2% effluent concentration, the proline content increased with increasing effluent concentration at all time periods. After 5 days of treatment, increase in proline content was 26.87, and 5.7% at 4, 6, and 8% effluent concentrations, respectively. However, it was 45.16, and 89.9% at 4, 6, and 8% effluent concentrations, respectively after 15 days. The present study shows that proline content was not effected upto 2% effluent concentration, but it increased with further increase in effluent concentration. It is well know that increase in proline content is associated with various biotic and abiotic stresses (Levitt, 1972). Thus, upto 2% effluent concentration does not induce any stress in water hyacinth. However, the plant is under stress at higher effluent concentration. This supports the fact that the water hyacinth yielded more phytoremediation at 2% effluent concentration as compared 286 Dr. Poonam Bathla

5 Increase(%) International Journal of Engineering Technology, Management and Applied Sciences to 4, 6 and 8% effluent concentration. The present study does not explain the mechanism of proline accumulation in water hyacinth in response to various effluent concentrations. It is known that proline biosynthesis in plants begins with glutamic acid. Steward et al. (1981) suggested that proline oxidation could function as a control mechanism for maintaining low cellular level of proline in turgid tissue, however, proline oxidation is reduced to negligible rates in stressed plants. According to them it seems likely that inhibition of proline oxidation is necessary in maintaining the high level of proline in stressed leaves. Thus, the accumulation of proline in response to different effluent concentration may be associated with the biosynthesis of proline from glutamic acid or/and due to inhibition of oxidation of proline into glutamic acid which is likely to be due to metal toxicity. Further studies are required to investigate the exact mechanism of proline accumulation in response to distillery effluent % 2% EC 4% EC % EC 8% EC Fig 4: Percentage Increase in Proline content of Water Hyacinth (E. crassipes) during phytoremediation of distillery effluent c. Metal Accumulation in Water Hyacinth The results of metal (iron, zinc, sodium, potassium, calcium and magnesium ) accumulation during phytoremediation of various concentrations of distillery effluent with in water hyacinth are presented in Fig 5 and 6. Concentration of iron, zinc, sodium, potassium, calcium and magnesium increased with in plant tissue with increasing time period at all the concentrations of distillery effluent. In the present study, it was found that the various heavy metals accumulated in plant tissue. The accumulation was more at 8% effluent concentration as compared to 6, 4, and 2% effluent concentration. Maximum accumulation of metals occurred at highest effluent concentration (8%) after 15 days. At 8% effluent concentration, the increase in iron, zinc, sodium, potassium, calcium and magnesium was 37.68, 29.35, 8.54,17.7, and 23.42%, respectively after 5 days; however, it was 62.8, and 32.17, 36.84, 21.5 and 27.31%, respectively after 15 days.the metal accumulation may be due to high uptake efficiency of water hyacinth due to its quest for nutrients. E. crassipes, has a property to accumulate heavy metals, such as cadmium, copper, lead and zinc in the root tissues of the plants. It is likely that water hyacinth contains metallothioneins (metal binding proteins) in response to heavy metal stresses (Kong and Tsang, 1998) which removes heavy metal by formation of chelates. 287 Dr. Poonam Bathla

6 Reduction (%) Reduction (%) International Journal of Engineering Technology, Management and Applied Sciences 35 Effluent concentration (2%) Fe Zn Na K Ca Mg Fig. 5: Metal Accumulation in Water Hyacinth (E. crassipes) during phytoremediation of (2%) Distillery Effluent Effluent concentration (8%) Fe Zn Na K Ca Mg 5 15 Time ( days) Fig. 6: Metal Accumulation in Water Hyacinth (E. crassipes) during phytoremediation of (8%) of Distillery Effluent The present study indicates E. crassipes can be successfully employed for reduction in metal content of effluent at lower effluent concentration however its reduction efficiency reduces at higher effluent concentration which can be directly related to the health of the plant and is reflected in terms of chlorophyll reduction and increase in free proline content of the water hyacinth. Conclusions : Based on present investigation, the aquatic weed E.crassipes an ideal floating macrophyte for phytoremediation of distillery effluent as it is highly efficient in terms of metal bioaccumulation. E.crassipes could be profitably used to remediate distillery effluent as it is widely available, fast growing, has simple growth and treatment setup requirements, high metal absorption capacities and ability to survive under adverse environmental conditions.phytoaccumulation with water hyacinth could be opted as a cost effective method for treatment of distillery effluent. References : Abbasi, S.A. and Ramasami, E Aquatic macrophytes in waste water treatment system: suitability, mechanism of action, design considerations economics and environmental impact. In: Biotechnological methods of pollution control. University Press (India) Ltd., Hyderabad, pp Dr. Poonam Bathla

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