Natural Zeolite Mediated Mercury Toxicity in Fish

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Asian J. Exp. Sci., Vol. 20, No. 2, 2006, 303-308 Natural Zeolite Mediated Mercury Toxicity in Fish M. K. Chaurasia and Subodh Kumar Jain* Department of Zoology Dr. H.S. Gour University SAGAR 470003 M.P. (India) Abstract : There is no truly effective treatment of mercuralism, once it gets in to the body. In the present investigation, efforts have been made to study the role of natural zeolite stilbite for remediation of mercury toxicity. Zeolites are naturally and artificially occurring ion exchangers. Heavy metal viz. mercury takes the position of element present in the molecular sieve of zeolite in the exposure water. Thus, heavy metal becomes unavailable to fish. During the study, experimental fish were exposed to mercuric chloride, mercuric chloride + stilbite and stilbite only. Values of protein contents in the liver, kidney and gills of teleost fish Heteropneustis fossilis in various experimental groups indicates that the toxicity of mercury is reduced due to addition of zeolite. It is also observed that exposure of fish to stilbite only increase the protein contents in the fish tissue significantly, suggests that natural zeolites can be used not only for the remediation of metal toxicity, rather also for enhancing protein contents in the fish. Key words : Stilbite, Mercury, Fish, Protein, Liver, Kidney, Gill. Introduction : According to both, the World Health Organization (WHO) and the US Environmental Protection Agency (EPA), consumption of fish and marine mammals, is the single most important source of human exposure to methyl mercury. Methyl mercury is a potent neurotoxin that concentrates at increasingly poisonous levels in the body. It can pass through the placental barrier during pregnancy causing severe neurological damage in developing fetuses. Fish dependent populations are not only exposed to mercury but their exposure is the most highly potent organic form of mercury that damages the central nervous system and can decimate fishing communities for generations. Mercury poisoning, also known as mercuralism, is the phenomenon of toxicities by contact with mercury. Human activities like the application of agricultural fertilizers and industrial waste water disposal are examples of how humans release mercury directly in to the soil and water. Mercury toxicity is a serious problem since it does not degrade in the environment; therefore its removal is difficult. It is highly toxic particularly to children and the developing foetus, where it interferes with development, particularly the maturation process of the brain. Whatever form, mercury is in elemental, inorganic or organic, it is toxic, and there is no truly effective treatment once it gets into the human body. Chelating therapy may help but there are doubts about the effectiveness. Fish accumulates mercury directly from food and the surrounding water which gets biomagnified in fish and fish products and constitutes an important part of human diet. * Corresponding author : Subodh Kumar Jain, Phone : +91-7582-237175 E-mail : subodhjain@sancharnet.in 303

Chaurasia M.K. and Jain S.K. (2006) Asian J. Exp. Sci., 20(2), 303-308 Methyl mercury is a well-known human neurotoxin. The accumulation of heavy metals in different organs of fish as a whole in marine as well as freshwater has been described by several workers (Eaton, 1974; Wright, 1976; Pentreath, 1977; Edgren and Notton, 1980). It has been demonstrated that gills and gut could be important transfer route for mercury in insect nymphs (Saouter et al., 1991 and Odin et al., 1995 and 1996). Mercury accumulates in fish tissue is usually in the form of methyl mercury, while the source is usually inorganic mercury (Huckabee et al., 1979). It is generally agreed that mercury concentrations in carnivorous fish are higher than in non carnivorous species (Watras and Huckabee, 1994). Schweiger (1957) investigated the effect of mercury ions on fish and their food organism and suggested a toxic threshold concentration of 0.03 mg/litre for various species tested. There is no truly effective treatment of mercuralism, once it gets in to the body. In the present investigation, efforts have been made to study the role of natural zeolite stilbite for remediation of mercury toxicity. Zeolites are naturally and artificially occurring ion exchangers. Heavy metal viz. mercury takes the position of element present in the molecular sieve of zeolite in the exposure water. Thus, heavy metal becomes unavailable to fish. Zeolites are naturally occurring, finegrained materials with pronounced ion exchange properties. Small amount of zeolites increase biomass production and improves nitrogenous efficiency. They promote better plant growth by improving the value of fertilizer. They also improve water quality and filtration in aquaculture. Element present within molecular sieve of zeolite is exchanged with heavy metal of water resource. This result into nonavailability of heavy metal to fish and thus deleterious effects to fish (Jain et al., 1997; Jain, 1999 and Shrivastava et al., 2004). Mercury causes depletion in protein, RNA and glycogen contents (Shakoori et al., 1994). In the contaminated water the sodium of zeolite is exchanged with heavy metal ions in their molecular sieve; at the same time, natural zeolite helps in protein digestion in cattle, sheep and goats (Petkova et al., 1984). The present study has been planned at assessing the protein contents of liver, kidney and gills of teleost fish Heteropneustes fossilis in normal conditions and under the influence of mercuric chloride, mercuric chloride+stilbite and only stilbite. This study explores the possibility of usefulness of natural zeolite stilbite for remediation of mercury toxicity and economic opportunities to fish farmers. Materials and Methods : Prior to experimentation, 120 healthy, adult specimens of teleost fish Heteropneustes fossilis (Bloch.) have been acclimatized for two weeks to laboratory conditions. By interpolation method, 96h LC 50 value for HgCl 2 determined (2 mgl -1 day -1 ). Fish were divided into four equal groups. Group I served as control while group II, III and IV exposed to sublethal concentration of mercuric chloride, mercuric chloride+stilbite and only stilbite, respectively. During experimental study, sublethal concentration of HgCl 2 and equal amount of stilbite powder sprinkled into exposure water. Fishes were sacrificed after 7, 14, 21, 28 and 35 days of exposure, their liver, kidney and gill tissues were removed 304

Natural Zeolite Mediated Mercury Toxicity Table : Values of protein content (mg/g) in the fish tissue in various treatment groups. Fish Days of Control Treatment groups tissue Exposure HgCl 2 HgCl 2 +Stilbite Only Stilbite 7 82.06±1.65 57.87±1.37 P<0.001 59.97±1.40 P<0.001 86.90±1.70 P<0.001 14 82.19±1.65 57.08±1.38 P<0.001 60.34±1.41 P<0.001 86.94±1.70 P<0.001 Liver 21 82.29±1.65 58.15±1.39 P<0.001 61.30±1.42 P<0.001 86.12±1.70 P<0.001 28 82.46±1.65 59.02±1.40 P<0.001 62.43±1.44 P<0.001 88.48±1.71 P<0.001 35 82.53±1.65 61.02±1.42 P<0.001 63.42±1.45 P<0.001 89.22±1.72 P<0.001 7 69.84±1.52 47.08±1.25 P<0.001 50.12±1.29 P<0.001 71.23±1.54 P<0.001 14 69.88±1.52 48.05±1.26 P<0.001 51.00±1.30 P<0.001 74.51±1.57 P<0.001 Kidney 21 69.89±1.52 49.04±1.27 P<0.001 52.20±1.31 P<0.001 75.21±1.58 P<0.001 28 69.98±1.52 50.15±1.29 P<0.001 53.38±1.33 P<0.001 76.42±1.59 P<0.001 35 69.98±1.52 51.48±1.31 P<0.001 54.72±1.35 P<0.001 78.06±1.61 P<0.001 7 63.93±1.46 44.70±1.22 P<0.001 47.67±1.26 P<0.001 64.85±1.47 P<0.001 14 63.85±1.46 45.68±1.23 P<0.001 48.48±1.27 P<0.001 69.50±1.52 P<0.001 Gill 21 63.98±1.46 46.43±1.24 P<0.001 49.81±1.28 P<0.001 70.02±1.52 P<0.001 28 64.00±1.46 47.09±1.25 P<0.001 50.73±1.30 P<0.001 71.03±1.53 P<0.001 35 64.02±1.46 48.81±1.27 P<0.001 51.70±1.31 P<0.001 72.06±1.54 P<0.001 305

Chaurasia M.K. and Jain S.K. (2006) Asian J. Exp. Sci., 20(2), 303-308 and processed for quantitative estimation of protein by Lowery (Folin-Ciocalteau) method. Student's t test was applied for statistical evaluation of data. Results and Discussion : According to observation table, values of protein contents in liver, kidney and gills are almost similar to those of the initial control fish. Sublethal concentration of mercuric chloride decreases the protein contents significantly. Addition of stilbite to the exposure water improves the values towards normal. When fishes were exposed to stilbite only, protein contents in the tissue increased in comparison to control. Ram and Sathyanesan (1984) reported that mercuric chloride reduces the protein, lipid and cholesterol contents in various tissues of Channa punctatus. Sastry and Gupta (1978) emphasized that over all decrease in protein contents is probably due to enzyme inhibition which plays an important role in protein synthesis. Kidney lysosomes tend to accumulate mercury (Suzuki, 1977). Although Backstrom (1969) reported that despite relatively high level of methyl mercury, fishes do not show any over toxicity. Dhanapakiam et al. (1998) studied the gill of mature Channa punctatus exposed to effluent of industrial water in the Cauvery river water for 45 days, revealed deformities and also observed that the secondary lamellae of primary filaments showed hyperplasia. Khangarot (1982) reported that gill epithelium-exhibited varying degrees of hypertrophy, the gap between the basement membrane and the epithelial layer increased in size and the lamellar capillary was dilated due to zinc toxicity in Puntius sophore. Liver is the most important target organ for heavy metal toxicity (Holocombe et al, 1976). Decrease in soluble protein, RNA and glycogen contents and hepatic enzymes due to mercury have been reported by Shakoori et al. (1994). There are a number of metal chelators which are used for the remediation of metal toxicity as reported by Graziano et al. (1985); Klavassen (1985); Chislom (1970 & 1971); Friedhein et al. (1978) and Hammond (1971). The element in the molecular sieve of zeolites is exchanged with metal ion thus the concentration of metal is decreased in the exposure water; as a result deleterious effects in the tissues are reduced (Jain, 1999, 2001; Jain and Shrivastava, 2000; Jain et al., 2003). The removal of ammonia from aqueous solution by natural zeolites has been investigated by Aral et al. (1999). Among the various cation exchangers, zeolites are preferred due to its high specificity for heavy metal cations (Sherman, 1978; Semmens and Sayfarth, 1978). In the present study, it can be concluded that natural zeolite stilbite is useful and can be used for attenuation of mercury toxicity and enhancing the protein contents in fish. References : Aral N., Gunay A., Serimoglu O., Cali M. and Debik E. (1999) : Ammonia removal from aqueous solution by ion exchange using natural zeolite. Fres. Environ. Bull. 8, 344-349. Backstrom J. (1969) : Distribution studies of mercuric pesticides in quail and some fresh water fishes. Acta Pharmacol. Toxicol. 27(3), 3-103. Chislom U. Jr. (1970) : Treatment of acute lead intoxication choice of chelating agents and supportive therapeutic measurements. Clin. Toxicol., 3, 527-540. Chislom U. Jr. (1971) : Treatment of lead poisoning, Mod. Treat. 8, 593-611. 306

Natural Zeolite Mediated Mercury Toxicity Dhanapakiam P., Ramasamy V.K. and Sompooranin (1998) : A study of the histopathological changes in gills of Channa punctatus in Cauvery river water. J. Environ. Bio., 19(3), 265-269. Eaton J. (1974) : Chronic cadmium toxicity to blue gill (Leposmis marco chritus rafinesque) Trans. Fish. Soc. 108, 729-735. Edgren M. and Notton M. (1980) : Cadmium uptake by fingerlings of perch (Pera flueriatits) studies by CD-115 nm at two different temperatures. Bull. Env. Contam. Toxicol. 24, 644-651. Friedhein E., Graziano J.H., Popovac D., Dragaric D. and Kaul B. (1978) : Treatment of lead poisoning by 2,3-dimercaptosuccinic acid. Lancet, 2, 1234-1236. Graziano J. H., Siris E. S., LoIacono N., Silverberg S.J. and Turgeon L. (1985) : 2, 3-dimercaptosuccinic acid as an antidote for lead intoxication Clin. Pharmacal. Ther., 37, 431-438. Hammond P.B. (1971) : The effect of chelating agents on the tissue distribution and excretion of lead. Taxicol Appl. Pharmacol., 18, 296-310. Holcombe G.W., Benoit D.A., Leonard E.N. and Mckim I.M. (1976) : Long term effects of lead exposure on three generations of brook trout Salvelinus fonlinalis. J. Fish Res. Board Can., 33, 1731-1741. Huckabee J.W., Elwood J.W. and Hildebrand S.G. (1979) : In : Accumulation of mercury in freshwater biota. (ed. Nriagu JO) : The biogeochemistry of mercury in the environment. p. 277-302. Jain S.K., Raizada A.K., Jain K. and Shrivastava S. (1997) : Protective role of zeolite on lead toxicity in freshwater fish. XII ISEB (21-26 Sept.) at Monopoli, Bari, Italy. Jain S.K. (1999) : Protective role of zeolite on short and long term lead toxicity in the teleost fish Heteropneustes fossilis, Chemosphere. 39(2), 247-251. Jain S.K. and Shrivastava S. (2000) : Zeolite mediated lead accumulation in fish tissue. Him. J. Environ. Zool., 14, 65-68. Jain S.K. (2001) : Zeolite influence on remediation of heavy metal toxicity in fish. Polish society of Humic substances, Grunwaldzka, 53, 350-357, Wroclaw, Poland, 77-78. Jain S.K. Shrivastava R.K. and Mishra M. (2003) : Chabazite mediated remediation of mercury toxicity in fish tissue: A biochemical study. 16 th international symposium on Environmental Biogeochemistry. Aomori prefecture, Japan Page 227. Khangarot B.S. (1982) : Histopathological changes in the branchial apparatus of Puntius sophore (Hamilton.) subjected to toxic doses of zinc Acta. Hydrobiol. 93(3), 352-358. Klavassen C.D. (1985) : Heavy metals and heavy metals antagonist. In: The Pharmacological basis of Therapeutics, (eds. Gilman, AG, Goddman, LS, Rail, TW, Murad F) Mc Millan, New York, p. 1605-1627. Odin M., Ribeyre F. and Boudou A. (1995) : Cadmium and methyl mercury bio- accumulation by nymphs of the burrowing mayfly Hexaginia rigida from the water column and sediment. Environ. Sci. Pollut. Res. 2, 145-152. Odin M., Ribeyre F. and Boudou A. (1996) : Temperature and ph effects on cadmium and methyl mercury bioaccumulation by nymphs of the burrowing mayfly Hexaginia rigida. Arch Environ. Toxicol. 31, 339-349. Pentreath R.J. (1977) : The accumulation of cadmium by the plaice Pleuronectes plastassa and the Thornbackary, Roja clavata, L. J. Exp. Mar. Biol. Ecol. 30, 223-232. Petkova E., Venkov T. and Chushkov D. (1984) : Cited in uses of natural zeolites by Alen Dyer paper presented at Symposium held in Manchester, April 1984. Ram R.N. and Sathyanesan A.G. (1984) : Mercuric chloride induced changes in the protein, lipid and cholesterol levels of liver and ovary of the fish Channa punctatus. Environ. Ecol. 2, 113-117. Saouter E. Lemenn R. Boudou A. and Ribeyre F. (1991) : Structural and ultra structural analysis of gills and gut of Hexagenia rigida nymphs in 307

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