Gülüzar ATLI, Esin GÜLNAZ CANLİ, Ali EROGLU, Zehra DOĞAN, Mustafa CANLI

Cadmium and Lead Alter the Antioxidant and Osmoregulation Systems in the Erythrocyte of Fish (Oreochromis niloticus)

Eritrositlerin tüm vücut hücrelerine oksijen taşımaları metabolizma için onları son derece hayati yapar. Metallerin kan yollarıyla taşınmaları eritrositlerin metallerle sürekli karşı karşıya kalması anlamındadır. Bu nedenle bu çalışma Nil çuprası (Oreochromis niloticus) eritrositlerinde akut (20µM, 2 gün) ve sub-kronik (10 µM, 20 gün) Cd ve Pb etkisi sonrası ozmoregülasyon (Na+/K+-ATPase, Mg+2-ATPase, Ca+2-ATPase) ve antioksidan sistem parametrelerinin (CAT, SOD, GPX, GR, GST, tGSH, rGSH, GSSG, GSH/GSSG) cevabını araştırmayı amaçlamaktadır. ATPaz'ların antioksidan sistem parametreleri ile karşılaştırıldığında daha duyarlı oldukları görülmüştür. Bütün ATPaz aktiviteleri akut ve sub-kronik metal etkilerinde önemli düzeyde artmıştır. Metal ve etki sürelerine bağlı olarak antioksidan sistem parametrelerinde farklılık olduğu ve Pb'nin daha çok etkiye neden olduğu gözlenmiştir. En çok GSH metabolizmasının özellikle de sub-kronik Pb etkisi sonrası değişmesine karşın CAT ve GPX aktiviteleri de azalmıştır. Akut Pb etkisi de azalan GPX ve artan GST aktivitelerine neden olmuştur. Akut ve sub-kronik Pb etkileri arasında GSH/GSSG oranı bakımından farklılıklar vardır. Cd, akut etki sonrası GSSG düzeyi ve GSH/GSSG oranındaki azalış ile sub-kronik etki sonrası GST aktivitesindeki azalış dışında antioksidan sistem parametrelerinde çok etkili olmamıştır. Bu çalışma, eritrosit osmoregülasyon ve antioksidan sistem parametrelerinin metal etkilerine duyarlı olduğunu ve stresle yüzleşen balıklar hakkında yararlı veriler sağlayabileceğini belirtmektedir.

Kadmiyum ve Kurşunun Balık (Oreochromis niloticus) Eritrositlerinde Antioksidan ve Ozmoregülasyon Sistemlerini Değiştirmesi

The erythrocytes deliver oxygen to all around body cells that make them extremely vital for the metabolism. Metals are transferred through the blood, meaning the erythrocytes face with metals continuously. The present study intended to explore the responses of osmoregulation (Na+/K+-ATPase, Mg2+-ATPase, Ca2+-ATPase) and antioxidant system parameters (CAT, SOD, GPX, GR, GST, tGSH, rGSH, GSSG, GSH/GSSG) in the erythrocytes of the Nile tilapia (Oreochromis niloticus) following acute (20 µM, 2 d) and sub-chronic (10 µM, 20 d) Cd and Pb exposures. ATPases were more sensitive compared to the antioxidant system parameters. All ATPase activities increased significantly in acute and sub-chronic metal exposures. Variations in antioxidant system parameters were recorded with metal and duration differences, Pb causing more alterations. GSH metabolism was mostly altered, especially after sub-chronic Pb exposure, though CAT and GPX activities decreased. Acute Pb exposure also caused a decrease in GPX activity and an increase in GST activity. There were differences in GSH/GSSG ratios between acute and sub-chronic Pb exposures. Cd caused few alterations in the antioxidant system parameters (GSSG and GSH/GSSG ratio decrease and GST increase). The present study demonstrated that the osmoregulation and antioxidant systems of the erythrocyte are sensitive to metal exposures and may provide useful data about the stress fish face.

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Akbarsha, M.A., Kadalmani, B., Girija, R., Faridha, A., Hamid, K.S. 2001. Spermatotoxic effect of carbendazim. Indian Journal of Experimental Biology, 39: 921-924.
Almeida, J.A., Diniz, Y.S., Marques, S.F.G., Faine, L.A., Ribas, B.O., Burneiko, R.C., Novelli, E.L.B. 2002. The use of the oxidative stress responses as biomarkers in Nile tilapia (Oreochromis niloticus) exposed to in vivo cadmium contamination. Environment International, 27: 673-679. doi:10.1016/S0160-4120(01)00127-1.
Alves, L.C., and Wood, C.M. 2006. The chronic effects of dietary lead in freshwater juvenile rainbow trout (Onchorhynchus mykiss) fed increased calcium diets. Aquatic Toxicology, 78: 217-232. doi:10.1016/j.aquatox.2006.03.005.
Atkinson, A., Gatemby, A.O., and Lowe, A.G. 1973. The determination of inorganic orthophosphate in biological systems. Biochimica et Biophysica Acta, 320: 195-204.
Atli, G., Alptekin, Ö., Tükel, S., and Canli, M. 2006. Response of catalase activity to Ag+, Cd2+, Cr6+, Cu2+ and Zn2+ in five tissues of freshwater fish Oreochromis niloticus. Comparative Biochemistry and Physiology, 143C: 218-224. doi:10.1016/j.cbpc.2006.02.003.
Atli, G., and Canli, M. 2007. Enzymatic responses to metal exposures in a freshwater fish Oreochromis niloticus. Comparative Biochemistry and Physiology, 145C: 282-287. doi: 10.1016/j.cbpc.2006.12.012.
Atli, G., and Canli, M. 2008. Responses of metallothionein and reduced glutathione in a freshwater fish Oreochromis niloticus following metal exposures. Environmental Toxicology and Pharmacology, 25: 33-38. doi:10.1016/j.etap.2007.08.007.
Atli, G., and Canli, M. 2010. Response of antioxidant system of freshwater fish Oreochromis niloticus to acute and chronic metal (Cd, Cu, Cr, Zn, Fe) exposures. Ecotoxicology and Environmental Safety, 73: 1884-1889. doi:10.1016/j.ecoenv.2010.09.005.
Atli, G., and Canli, M. 2011a. Essential metal (Cu, Zn) exposures alter the activity of ATPases in gill, kidney and muscle of tilapia Oreochromis niloticus. Ecotoxicology, 20: 1861-1869. doi: 10.1007/s10646- 011-0724-z.
Atli, G., and Canli, M. 2011b. Alterations in ion levels of freshwater fish Oreochromis niloticus following acute and chronic exposures to five heavy metals. Turk Journal of Zoology, 35: 725-736. doi:10.3906/zoo- 1001-31.
Atli, G., Yuzbasioglu Ariyurek, S., Kanak, E.G., and Canli, M. 2015. Alterations in the serum biomarkers belonging to different metabolic systems of fish (Oreochromis niloticus) after Cd and Pb exposures. Environmental Toxicology and Pharmacology, 40: 508-515. doi:10.1016/j.etap.2015.08.001.
Barata, C., Varob, I., Navarro, J.C., Arun, S., and Porte, C. 2005. Antioxidant enzyme activities and lipid peroxidation in the freshwater cladoceran Daphnia magna exposed to redox cycling compounds. Comparative Biochemistry and Physiology, 140C: 175-186. doi:10.1016/j.cca.2005.01.013.
Baysoy, E., Atli, G., and Canli, M. 2013. The Effects of Salinity and Salinity+Metal (Chromium and Lead) Exposure on ATPase Activity in the Gill and Intestine of Tilapia Oreochromis niloticus. Archives of Environmental Contamination and Toxicology, 64: 291-300. doi:10.1007/s00244-012-9825-9
Bergmeyer, H.U. 1974. Methods of enzymatic analysis. Academic press, New York.
Canli, M., and Stagg, R.M. 1996. The effects of in vivo exposure to cadmium, copper, and zinc on theactivities of gill ATPases in the Norway lobster Nephrops norvegicus. Archives of Environmental Contamination and Toxicology, 31: 491-501. doi: 10.1007/BF00212433.
Cao, L., Huang, W., Liu, J., Yin, X., and Dou, S. 2010. Accumulation and oxidative stress biomarkers in Japanese flounder larvae and juveniles under chronic cadmium exposure. Comparative Biochemistry and Physiology, 151C: 386-392. doi:10.1016/j.cbpc.2010.01.004.
Carlberg, I., and Mannervik, B. 1975. Purification and characterization of the flavoenzyme glutathione reductase from rat liver. The Journal of Biological Chemistry, 250: 5475-5480.
Congleton, J.L., and La Voie, W.J. 2001. Comparison of blood chemistry values of samples collected from juvenile chinook salmon by three methods. Journal of Aquatic Animal Health, 13: 168-172. doi:10.1577/1548-8667(2001)013_____0168:COBCVF>2.0.CO;2.
De la Torre, F.R., Salibian, A., and Ferrari, L. 2000. Biomarkers assessment in juvenile Cyprinus carpio exposed to waterborne cadmium. Environmental Pollution, 109: 277-282. doi:10.1016/S0269-7491(99)00263-8.
Dimitrova, M.S.T., Tsinova, V., and Velcheva, V. 1994. Combined effect of zinc and lead on the hepatic superoxide dismutase-catalase system in carp (Cyprinus carpio). Comparative Biochemistry and Physiology, 108C: 43-46.
Eroglu, A., Dogan, Z., Kanak, E.G., Atli, G., and Canli, M. 2015 Effects of heavy metals (Cd, Cu, Cr, Pb, Zn) on fish glutathione metabolism. Environmental Science and Pollution Research, 22: 3229-3237. doi: 10.1007/s11356-014-2972-y.
Gabryelak, T., Filipiak, A., and Brichon, G. 2000. Effects of zinc on lipids of erythrocytes from carp (Cyprinus carpio L.) acclimated to different temperatures. Comparative Biochemistry and Physiology, 127C: 335-343. doi:10.1016/S0742-8413(00)00161-4
Griffith, O.W. 1980. Determination of glutathione and glutathione disulfide using glutathione reductase and 2-Vinylpyridine. Analytical Biochemistry, 106: 207- 212. doi:10.1016/0003-2697(80)90139-6.
Grosell, M., McDonald, M.D., Walsh, P.J., and Wood, C.M. 2004. Effects of prolonged copper exposure in the marine gulf toadfish (Opsanus beta) II: Copper accumulation, drinking rate and Na+-K+ ATPase activity in osmoregulatory tissues. Aquatic Toxicology, 68: 263-275. doi:10.1016/j.aquatox.2004.03.007.
Habig, W.H., Pabst, M.J., and Jakoby, W.B. 1974. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. Biological Chemistry, 249: 7130-7139.
Heath, A.G. 1987. Water Pollution and Fish Physiology. CRC Press, Florida, 245 pp.
Jorgensen, S.W. 2012. Ecotoxicology: a derivative of encyclopedia of ecology. Academic Press, London.
Larsson, A., Haux, C., Sjobeck, M.L. 1985. Fish physiology and metal pollution: Results and experiences from laboratory and field studies. Ecotoxicology and Environmental Safety, 9: 250-281. doi:10.1016/0147- 6513(85)90045-4.
Lima, P.L., Benassi, J.C., Pedrosa, R.C., Dal Magro, J., Oliveira, T.B., Wilhelm Filho, D. 2006. Time-course variations of DNA damage and biomarkers of oxidative stress in tilapia (Oreochromis niloticus) exposed to effluents from a Swine industry. Archives of Environmental Contamination and Toxicology, 50: 23-30. doi: 10.1007/s00244-004-0178-x.
Livingstone, D.R., Lips, F., Martinez, P.G., and Pipe, R.K.1992. Antioxidant enzymes in the digestive gland of the common mussel Mytilus edulis. Marine Biology, 112: 265-276. doi: 10.1007/BF00702471.
Lowry, O.H., Rosebrough, N.J., Farra, N.J., and Randall, R.J. 1951. Protein measurements with the folin phenol reagent. The Journal of Biological Chemistry, 193: 265-275.
Mance, G. 1987. Pollution threat of heavy metals in aquatic environment. Elsevier, London, 126 pp.
Marcon, J.L., Filho, D.W. 1999. Antioxidant processes of the wild tambaqui, Colossoma macropomum (Osteichthyes, Serrasalmidae) from the Amazon. Comparative Biochemistry and Physiology, 123C: 257-263.
McCord, J.M., and Fridovich, I. 1969. Superoxide dismutase: An enzymatic function for erythrocuprein (hemocuprein). The Journal of Biological Chemistry, 244: 6049-6055.
McGeer, J.C., Szebedinszky, C., McDonald, D.G., and Wood, C.M. 2000. Effects of chronic sublethal exposure to waterborne Cu, Cd or Zn in rainbow trout. 1: Iono-regulatory disturbance and metabolic costs. Aquatic Toxicology, 50: 231-243. doi:10.1016/S0166-445X(99)00105-8.
Mills, G.C. 1959. The Purification and Properties of Glutathione Peroxidase of Erythrocytes. The Journal of Biological Chemistry, 234: 502-506.
Min, E.Y., and Kang, C.J. 2008. Effect of waterborne benomyl on the hematological and antioxidant parameters of the Nile tilapia, Oreochromis niloticus. Pesticide Biochemistry and Physiology, 92: 138-143. doi:10.1016/j.pestbp.2008.07.007
Nagalakshmi, N., and Prasad, M.N.V. 1998. Copperinduced oxidative stress in Scenedesmus bijugatus: Protective role of free radical scavengers. Bulletin of Environmental Contamination and Toxicology, 61: 623-628.
Oner, M., Atli, G., and Canli, M. 2008. Changes in serum biochemical parameters of freshwater fish Oreochromis niloticus following prolonged metal (Ag, Cd, Cr, Cu, Zn) exposures. Environmental Toxicology and Chemistry, 27: 360-366.
Orbea, A., Fahimi, H.D., and Cajaraville, M.P. 2000. Inmunolocalization of four antioxidant enzymes in digestive glands of mollusks and crustaceans and fish liver. Histochemistry and Cell Biology, 114: 393-404. doi 10.1007/s004180000207.
Parvez, S., Sayeed, I., and Raisuddin, S. 2006. Decreased gill ATPase activities in the freshwater fish Channa punctata (Bloch) exposed to a diluted paper mill effluent. Ecotoxicology and Environmental Safety, 65: 62-66. doi:10.1016/j.ecoenv.2005.07.010.
Pena-Llopis, S., Pena, J.B., Sancho, E., Fernandez-Vega, C., and Ferrando, M.D. 2001. Glutathione-dependent resistance of the European eel Anguilla anguilla to the herbicide molinate. Chemosphere, 45: 671-681. doi:10.1016/S0045-6535(00)00500-2.
Pinto, E., Sigaud-Kutner, T.C.S., Leitao, M.A.S., Okamoto, O.K., Morse, D., and Colepicolo, P. 2003. Heavy metal-induced oxidative stress in algae. Journal of Phycology, 39: 1008-1018. doi: 10.1111/j.0022- 3646.2003.02-193.x.
Ramos-Vasconcelos, G.R., and Hermes-Lima, M. 2003. Hypometabolism, antioxidant defenses and free radical metabolism in the pulmonate land snail Helix aspersa. Journal of Experimental Biology, 206: 675- 685. doi: 10.1242/jeb.00124.
Roche, H., and Bogé, G. 1996. Fish blood parameters as a potential tool for identification of stress caused by environmental factors and chemical intoxication Marine Environmental Research, 41: 27-43. doi:10.1016/0141-1136(95)00015-1.
Ruas, C.B.G., Carvalho, C.S., Araujo, H.S.S., Espindola, E.L.G., and Fernandes, M.N. 2008. Oxidative stress biomarkers of exposure in the blood of cichlid species from a metal-contaminated river. Ecotoxicology and Environmental Safety, 71: 86-93. doi:10.1016/j.ecoenv.2007.08.018.
Rudneva, I.I., Skuratovskaya, E.N., Dorohova, I.I., and Kovyrshina, T.B. 2012. Use of fish blood biomarkers for evaluating of marine environment health. World Journal of Science and Technology, 2: 19-25.
Sanchez, W., Palluel, O., Meunier, L., Coquery, M., Porcher, J.M., and Ait-Aissa, S. 2005. Copperinduced oxidative stress in three-spined stickleback: relationship with hepatic metal levels. Environmental Toxicology and Pharmacology, 19: 177-183. doi:10.1016/j.etap.2004.07.003.
Saglam, D., Atli, G., Dogan, Z., Baysoy, E., Gurler, C., Eroglu, A., and Canli, M. 2014. Response of the Antioxidant System of Freshwater Fish (Oreochromis niloticus) Exposed to Metals (Cd, Cu) in Differing Hardness. Turkish Journal of Fisheries and Aquatic Sciences, 14: 43-52. doi: 10.4194/1303-2712- v14_1_06.
Sevgiler, Y., Piner, P., Durmaz, H., and Uner, N. 2007. Effects of N-acetylcysteine on oxidative responses in the liver of fenthion exposed Cyprinus carpio. Pesticide Biochemistry and Physiology, 87: 248-254. doi:10.1016/j.pestbp.2006.08.003.
Watson, T.A., and Beamish, F.W. 1981. The effects of zinc on branchial adenosine triphosphatase enzymes in vitro from rainbow trout, Salmo gairdneri. Comparative Biochemistry and Physiology, 68C: 167-173.
Woo, S.P.S., Liu, W., Au, D.W.T., Anderson, D.M., andWu, R.S.S. 2006. Antioxidant responses and lipidperoxidation in gills and erythrocytes of fish (Rhabdosarga sarba) upon exposure to Chattonella marina and hydrogen peroxide: Implications on the cause of fish kills. Journal of Experimental Marine Biology and Ecology, 336: 230-241. doi:10.1016/j.jembe.2006.05.013.
Zikic, R.V., Stajn, A.S., Pavlovic, S.Z., Ognjanovic, B.I., and Saicic, Z.S. 2001. Activities of superoxide dismutase and catalase in erythrocytes and plasma transaminases of gold fish (Carassius auratus gibelio Bloch.) exposed to cadmium. Physiological Research, 50: 105-111.