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International Journal of Biotechnology (IJBT), Volume 1, Issue 1, January- March 2012Unlu E 2004). Usually among aquatic animals the bivalves which are filter feeders have verygood ability in accumulating large quantities of heavy metals and microbes mainlypathogens. The dumping of enormous of quantities of sewage and industrial effluents in toestuaries has resulted in a drastic reduction of shallow water fish populations, increasedpollution and ecological imbalance resulting in the large scale is appearance of numerousflora and fauna ( Rajendran et al .,2004).Aquatic pollution started long back ,but intensifiedduring the last few decades and now the situation has become alarming especially in India (Girija et al ,2007) .The distribution of heavy metals in solution has widely been recognisedas a major factor in the geochemical behaviour, transport and biological effects of theseelements in natural waters ( Ananthan et al.,1992: Karthikeyan et al.,2004,2007). Ennorecreek is situated in northeast coast of Chennai city, Tamilnadu India. Ennore comprises oflagoons, with salt marshes and backwaters, which are submerged under water during hightide and form an arm of the sea opening in to the Bay of Bengal. The total area of the creekis 2.25 sq km and is nearly 400 meter wide. Its channels connect it to the Pulicat lake to thenorth and to the Kortalaiyar river in the south ( Kannan et al., 2007).Ennore receives sewagefrom in and around Ennore area, Royapuram and industrial effluents from Manali. Thedredging activities in Ennore area result in changes in the landscape, Sediment transport, anddust pollution to the coast by quarrying process .Bioaccumulation of heavy metals affect theorganisms by accumulating in their tissues and are transferred to the next trophic level. Theaccumulated heavy metal becomes toxic when it crosses the permissible limit. Accumulationof heavy metal depends on various factors like concentration of metals in water and exposureperiod; although some other environmental factors such as salinity ,pH , hardness andtemperature play significant roles in metal accumulation. (Blackmore and Wang,2003).Mussels have been considered as potential biomonitor for metallic contamination inmarine ecosystem (Jung and Zauke,2008 ). Exposure to heavy metals can also affectreproduction efficiency of aquatic biota and can lead to gradual extinction of theirgenerations in polluted waters (Sridhara et al., 2008). Hence an study has been conducted toevaluate the heavy metals like Zn, Cd, Pb and Cu in the tissues of Perna viridis and in thewater.MATERIALS AND METHODSSample preparation for metal analysis The mussel tissue was weighed , dried at 60°c in an oven until they reached a constantweight. The analysis of trace metals was carried out using the standard method ( Alam et al.,2002). All the reagents used are of analytical grade. The sample was digested withconcentrated nitric acid. The samples were transferred to a clean beaker. Then 10ml ofconcentrated nitric acid was added and the sample was heated using a hot plate , continuinguntil every sample was completely digested. After cooling a further 10 ml of 1N nitric acidwas added. The solution was then diluted and filtered through whatman filter paper or0.45µm nitrocellulose membrane filter. Determination of the elements in all samples wascarried out by ICP-AES (optima 2100 DV, Perkin Elmer, USA). 9
International Journal of Biotechnology (IJBT), Volume 1, Issue 1, January- March 2012Analysis of water sample Water samples were filtered through 0.45µm whatmann no.1 filter paper and thenwater sample was adjusted to pH 3.5 with HCl. The solution was transferred to a separatoryfunnel. 10ml of a freshly prepared 2% solution of amino-pyrolidine dithiocarbanate (APDC)was added and the mixture was shaken by a mechanical shaker, 20ml of methyl isobutylketone was added and the mixture was again shaken for 2 minutes. The phase was separatedand analysed using ICP-AES (Optima 2100 DV, Perklin- Elmer ,USA). A blankdetermination was done using the same procedure but without water samples.RESULTS AND DISCUSSIONTable 1: Bioaccumalation of heavy metals (Cadmium, Zinc ,Lead, Copper and (ppm) inperna viridis from Ennore creek during pre monsoon, monsoon, post monsoon and summerS.No Heavy metal Post Summer Pre monsoon Monsoon Monsoon1 Cd 0.0087 0.0065 0.0078 0.00562 Zn 2.132 - 0.087 0.0843 Pb 0.097 0.054 0.353 0.3474 Cu 0.376 - 0.324 0.364Table 2: Bioaccumalation of heavy metal (Cadmium, Zinc ,Lead, Copper and (µg/ml ) inwater from Ennore creek during pre monsoon, monsoon, post monsoon and summerS.No Heavy metal Post Summer Pre monsoon Monsoon Monsoon1 Cd 4.370 3.532 2.457 3.7892 Zn 35.93 28.43 19.12 26.733 Pb 8.342 7.365 7.023 7.8724 Cu 153.073 176.732 132.33 151.22 The result on heavy metals observed during present study in Ennore estuary (June2009 to June 2010) is given in Table 1 and Table 2. The overall heavy metal accumulation can be presented in descending order asfollows. In mussel sample Zn<Cu<Pd<Cd , In water sample Cu<Zn<Pd<Cd In the present study higher pollution of Copper and Zinc was found in Ennore area.Heavy metals were maximum during post monsoon and summer seasons followed bymonsoon and minimum during pre monsoon season. In the Ennore seacoast theconcentrations of metals were observed to be higher during summer than the monsoon. Theseseasonal low values may be attributed to fresh water input following rain as well as due to the 10
International Journal of Biotechnology (IJBT), Volume 1, Issue 1, January- March 2012release of surplus water from the Poondi reservoir in to the sea, while the higher values insummer are due to the evaporation of water, raising metal concentrations. In an earlier study, lower metal concentrations were observed during winter andhigher during the summer season. (Caccia and Millero,2003).Our study reveals high levels ofZinc and Lead compared to Copper and Cadmium. Concentrations of heavy metals were notuniform in water. Copper was high during summer season. Zinc, Cadmium and Lead weremore in post monsoon season.Perna viridis accumulated heavy metal Zinc at maximum andheavy metal cadmium at minimum. The data obtained in the present study in green musselsshows that metal Zinc is accumulated in higher concentration than cadmium, Zinc, lead andCopper. And in water Copper is higher . There is no correlation between the metals from thepresent data.Some authors have sought to establish correlations between various metals butthe scientific views and interpretations continue to be at variance (Bruland and Franks, 1983).Kang, Choi oh, Wright and Koh (1999) who worked on Asian periwinkle ( Littorinabrevicula) noticed that certain toxic metals, Cadmium and Lead in the tissue reflectedenvironmental levels ,Whereas Copper and Zinc were regulated by marine gastropods. Thisphenomenon was also seen in other species of the periwinkle by Young (1975) Bryan et al.,1983, Langston and Zhole (1986), Marigomiz and Ireland (1989,1990).Zinc is well regulatedin crustaceans (Bryan,1967) with excess metals stored in the hepatopancreas or excreted. (Colvocoresses and Lynch 1975).The total metabolic requirement for zinc in marine decapodscrustaceans is 71 µg/g dry weight (White and Rainbow,1985).Cadmium is a non essentialelement and is not well regulated in decapods crustaceans (Bryan ,1979; Rainbow,1985,Wong and Rainbow,1986).Metals such as Fe, Cu, Zn and Mn are essential metals as theyplay an important role in biological systems ,Hg, Cd and Pb are non- essential and are toxictoo. Lead in water comes from industrial ,mines and smelter discharges before beingdeposited in the sediment sinks. Lead is also used in the production of lead acid batteries,solder, alloys ,cable sheathi ng, pigments, rust inhibitors, ammunition, glazes and plasticstabilizers. Prabhahar et al., (2010). Lead toxicity leads to anaemia both by impairment ofhaemo – biosynthesis and acceleration of red blood cell destruction. The high metalconcentration may be originated from anthropogenic sources such as waste incineration,vehicle operations, combustible consumptioned fertilizer use, Which likely come from theupper basin of the lotic system that flow in to the wetland .The great difference in metalconcentration between low and high water period should be taken in to account whenanalysing contamination in waters .It is clear that water quality has seasonal variation, factthat should be considered where seafood plays an important role. Heavy metal in to coastalwaters need to be examined for its impact on the ability of marine organisms to survive andreproduce. Evaluation of important and valuable mussels is necessary prior to the futuredevelopment of industries or power plants near the marine environment. A realisticassessment of metals as toxicants in man’s environment is essential to establish meaningfulpollution guidelines. 11
International Journal of Biotechnology (IJBT), Volume 1, Issue 1, January- March 2012REFERENCES 1. Alam ,M.G.M., Tanaka, A., Allinson , G., Stagni , F. and Snow, E., 2002. A comparison of trace element concentration in cultured and wild carp. Cyprinus carpio of lake Kasumigaura , Japan . Ecotoxicol . Environ .Saf., 53: 238-243. 2. Ananthan ,G., P.Sampathkumar, P.soundarapandiyan and L.Kannan : 2005 Heavy metal concentration in Ariyankuppam estuary and Verampattinam coast of Pondicherry. Indan J. Fish., 52,501 -506. 3. Ananthan, G., M. Ganesan, P. Sampathkumar, M. Matheven pillai and L. Kannan: 1992 Distribution of trace metals in water , sediment and plankton of the velar estuary. Seaweed Res .Utiln., 15,69-75. 4. Ananthan,G., M.Ganesan,P. Sampath kumar,M.Mathevan pillai and L.Kannan: 1992 Distribution of trace metals in water, sediment and plankton of the Vellar estuary seaweed Res.utiln., 15,69 -75. 5. Bruland ,K.W and Franks , R.P., 1983.Mn,Ni,Cu,Zn and Cd in the western North Atlantic.In : Trace metals in sea water ,et al. (Ed) Nato.series .Plenum press. Pp.395- 414. 6. Bryan ,G.W., 1979. Bioaccumulation of marine pollutants Philosophical Transactions of the Royal Society London series, B286 : 483-505. 7. Bryan ,G.W., Langston ,W.J., Hummerstone , L.G., Burst, G.R and Ho,Y.B1983. An assessment of the gastropod, Littorina littorea, as an indicator of heavy metal contamination in United kingdom estuaries. Journal of Marine Biological Association of UK,63: 327-345. 8. Bryan, G .W., 1976. Heavy metal contamination in the sea In: Marine pollution ,Johnston , R. (Ed) ,Academic press London .P. 185 – 302. 9. Bryand,G.W.,1984 Pollution due to heavy metals and their compounds.In: Marine e cology (Ed.o.Kinne) Vol v.Part 3, John wiley and sons,New york.pp 1289-1432 10. C.Prabhahar, K.saleshrani and R.Enbarasan .,2011. Seasonal variations of heavy metals distribution in water and sediments of Kadalur, coastal zone, Tamilnadu, India. Indian j. of Recent Scientific Research.,Vol.1 Issue ,3,pp.91-95,March 2011. 11. Caccia , V.G and Millero , F.J., 2003. The distribution and seasonal variation of dissolved trace metals in the Florida bay and adjacent waters. Aqua – Geochem., 9: 111- 144. 12. Colvocoresses , J.A and Lynch ,M.P., Variations in serum constituents of the blue crab, Callinectes sapidus : Copper and Zinc, Comparititive Biochemistry and Physiology , 50 A: 135 – 139. 12
International Journal of Biotechnology (IJBT), Volume 1, Issue 1, January- March 2012 13. Girija, T.R., Mahanta, C and Chandramouli, V.2007. Water quality assessment of an untreated effluent imparted urban stream: The Bharalu Tributary of the Brahmaputra River; India, Environ, Monitor ,Assess.,130:221-236 14. Jung ,K and Zauke ,G .p 2008. Bioaccumulation of trace metals in the brown shrimp Crangon crangon (Linnaeus,1758) from the German Wadden sea. Aquat. Toxicol., 88: 243-249 15. Jung,K and Zauke ,G.p 2008. Bioaccumulation of trace metals in the brown shrimp Crangon crangon (Linnaeus,1758) from the German Wadden sea. Aquat. Toxicol., 88: 243-249 16. Kannan , K .s., Lee, K J .Krishnamoorthy , R . Purusothaman , A. Shanthi, K and Rao, R.2007. Aerobic chromium reducing Bacillus cereus isolated from the heavy metal contaminated Ennore creek sediment, North of Chennai, Tamilnadu, South east India .Res .J. Microbiol.,2 (2):130-140. 17. Kannan, K .s., Lee, K J .Krishnamoorthy, R.Purusothaman, A. Shanthi, K and Rao, R.2007. Aerobic chromium reducing Bacillus cereus isolated from the heavy metal contaminated Ennore creek sediment, North of Chennai, Tamilnadu, South east India .Res .J. Microbiol.,2 (2):130-140. 18. Karadede H, Oymsak S A, Unlu E(2004) .Heavy metals in mullet ,Liza abu and catfish, Silurus triostegus, from the Ataturk dam lake(Euphrates) Turkey. Environ .Int 30: 183-188 19. Karthikeyan R., S.Vijayalakshmi and T. Balasubramanian,2004. Seasonal distribution of heavy metals in the sediments from Uppanar estuary (East coast of India) J. Aqua bioll., 19, 119-122. 20. Karthikeyan R., S.Vijayalakshmi and T. Balasubramanian,2004. Seasonal distribution of heavy metals in the sediments from Uppanar estuary (East coast of India) J. Aqua bioll., 19, 119-122. 21. Karthikeyan, R., S. Vijayalakshmi and T. balasubramanian: 2007.Monthly variations of heavy metals and metal resistant bacteria from the Uppanar estuary (South east coast of India ). Res .J Microbiol.,2 ,50-57. 22. Langston ,W.J and Zhou, M .,1986.Evaluation of the significance of metal binding proteins in the gastropod, Littorina littorea, Marine Biology, 92: 505 – 515. 23. Leland H V, Kuwahara J S (1985).Trace metals in Raul GM, petrocelli SR (2nd eds )Fundamentals of aquatic toxixicity , Hemisphere pub. corp . Newyork . pp. 374-415. 24. Marigomez , I and Ireland, M.P., 1989.Accumulation , distribution and loss of Cadmium in the marine prosobranch , Littorina littorea (L ) science of the environment. 13
International Journal of Biotechnology (IJBT), Volume 1, Issue 1, January- March 2012 25. Marigomez , I and Ireland, M.P., 1990. Laboratory study of Cadmium exposure in Littorina littarea in relation to environmental Cadmium and exposure time science of the Total Environment, 90 : 75-87. 26. Pillai, Mathevan M.,1994.Hydrobiological investigations on the inter- tidal diatoms of the Cuddalore – Uppanar estuary (India),Ph.D., Thesis, Annamalai Unversity ,India,p.86. 27. Prabhahar .C., Saleshrani .K .,and Enbarasan.R seasonal variations of heavy metals distribution in water and sediments of Kadalur, coastal zone ,Tamilnadu, India, vol 1, Issue , 3, pp 91-95. 28. Pragatheeswaran,V., B. Loganathan , A.Ramesh and V.K Venugopalan:1986 Distribution of heavy metals and organic carbon in sediments off Madras and Vishakpattinam.Mahasagar- bull.Nat.Inst .Oceanogr.,19,39-44. 29. Rainbow P.S , 1985.The biology of heavy metals in the sea .International Journal of Environmental studies, 25 : 195 – 211. 30. Rajendran , N., Baskarasanjeevi , S., Ajmalkhan S and Balasuramanian T(2004). Ecology and biodiversity of Eastern ghats – Estuaries of India. EPTRI – Envis News letter,10,1-11. 31. Rajkumar,J.S.I ,John Milton,M.C and Arockia Rita, J.J. Assesment of Heavy metals (Cd,Cu, Pb and Zn )in Mugil cephalus and Penaeus indicus and meriterix meriterix of the Ennore creek, Tamilnadu,India. 32. Ridgway, J and Shimmield, G.2002.Estuaries as repositories of historical contamination and their impact on shelf seas .Estua.coas. shelf sci., 55:903-928 33. Ridgway, J and Shimmield, G.2002.Estuaries as repositories of historical contamination and their impact on shelf seas .Estua.coas. shelf sci., 55:903-928 34. Senthilnathan,S and T.Balasubramanian, 1997.Distribution of heavy metals in estuaries of south east coast of India.Indian J Mar sci.,26,95-97. 35. Sridhara, C.N., Kamal, C.T and Raj, S.S.D.2008. Assesing risk of heavy metals from consuming food grown on sewage irrigated soils and food chain transfer. Ecotoxicol. Environ.Saf.,69(3):513-524. 36. Sridhara, C.N., Kamal, C.T and Raj, S.S.D.2008. Assesing risk of heavy metals from consuming food grown on sewage irrigated soils and food chain transfer. Ecotoxicol. Environ.Saf.,69(3):513-524. 37. White S.L and Rainbow , P.S, 1985 On the metabolic requirements for Copper and Zinc in Molluscs and Crustaceans. marine Environmental Research, 16: 215 – 229. 38. Wong V.W.T and Rainbow ,P.S 1986. Apparent and real variability in the presence and metal contents of metallothioneins in the crab Carcinus maenas including the effects of isolation,procedure and metal induction . comp. Biochem. Physiol., 83A :157-177 39. Young , M.L., 1975. The transfer of Zn 65 and Fe 59 along a Fucus serratus (L.) Littoria obstusata food chain. J .Mar. Biol. Assoc.UK.55: 583 14