Haematotoxicity of Cypermethrin to African Catfish Clarias ...

JOURNAL OF ENVIRONMENTAL ENGINEERING AND TECHNOLOGY VOL. 1, NO. 2, MARCH 2012 20Haematotoxicity of Cypermethrin to AfricanCatfish Clarias Gariepinus under LaboratoryConditions1* Akinrotimi, O.A 2 Gabriel, U.U and 3 Ariweriokuma, S.VAbstract: Juveniles of Clarias gariepinus (meanweight 276.443±81.91SD and mean length29.47cm±1.88SD were exposed (three fish PC tank)to 0.00 (control), 0.05, 0.10, 0.20 and 0.25ppmcypermethrine solution in triplicates in rectangularglass aquaria for 10 days to determined the effect ofthe exposure on some haematological parameterssuch as Haemoglobin (Hb), Packed Cell Volume(PCV), leucocrit, White Blood Cell (WBC),differential counts (neutrophils, lymphocytes,eosinophils and monocytes), erythrocytesedimentation rate (ESR), Thrombocytes, RedBlood Cell (RBC) and its indices Mean CorpuscularHaemoglobin (MCH), Mean CorpuscularHaemoglobin Concentration (MCHC), and MeanCorpuscular Volume (MCV). The results from thestudy indicated a significant (P

JOURNAL OF ENVIRONMENTAL ENGINEERING AND TECHNOLOGY VOL. 1, NO. 2, MARCH 2012 21al., 2006), Monocrotophous (Yaji and Auta,2007), Kerosene (Gabriel et al., 2007). Thesestudies showed that various degrees ofhaematological changes could be produced byexposure of C. gariepinus to different levels oftoxicants under laboratory conditions.Despite a number of studies on the effects oftoxicants on haematology of C. gariepinus littleis known on the haematological changes that C.gariepinus may suffer under exposure tocypermethrin, a pesticide commonly applied inagriculture operations. The present study wascarried out to assess the effect of exposure of C.gariepinus to cypermethrin on its haematologicalparameters under laboratory conditions.II. MATERIALS AND METHODSA total of 18 Clarias gariepinus (mean bodyweight 276.44g ±8190SD and mean total length29.47cm±1.88SD) were obtained from a privatefarm of Abuloma Town in Port Harcourt LocalGovernment Area of Rivers State in Nigeria.They were then transported in plastic aquaria tothe fisheries research laboratory of the RiversState University of Science and Technology, PortHarcourt, Nigeria. The fish were acclimated tolaboratory condition for a period of one week.During the exposure period, the fish were fed oncommercial diet (35%CP) once a day at 3% oftheir body weight.After the period of acclimation, the fish wererandomly distributed into eighteen glass aquaria(56 x 28 x 28cm) with 30 litres of water at threefish per aquarium. Each of the aquaria containedeach of the following concentrations ofcypermethrin: 0.05, 0.10, 0.15, 0.20 and 0.25ml/l. The control group of fish were kept inaquaria devoid of toxicant (0.00mlL -1 ) each ofthese treatments had three replicates. During theexposure period which lasted for 10days, somewater quality parameters namely pH,conductivity, dissolved oxygen, alkalinity andhardness were measured thrice using the methodsdescribed by APHA (1985). Fresh preparation ofthe test solution were introduced into theexperimental media on daily basis.At the end of the experiment, blood wascollected from the fish by cardiac puncture usinga 21 – gauge hypodermic needle and syringe.The following haematological parametershaemoglobin (Hb), Packed Cell Volume (PCV),Red Blood Cell (RBC) and ErythrocyteSedimentation Rate (ESR), were determinedusing the methods described by Blaxhall andDaisley (1973). Differential counts (neutrophils,lymphocytes, eosinophils, monocytes) andleucorit were evaluated according to McLeay andGordon (1977). Red blood indices such as meancorpuscular haemoglobin concentration (MCHC),mean corpuscular haemoglobin (MCH), andMean Corpuscular Volume (MCV) werecalculated using the formula of Seiverd (1983).Data obtained from the study were analyzed bya one-way analysis of variance (ANOVA) test at0.05% probability level and differences amongmeans were indicated using Tukey test (Zar,1984).III. RESULTSThe water quality parameters in the varioustreatment levels (Table 1) showed that there wereno significant differences between the treatmentand the control. The haematological changesproduced by the effects of exposure of C.gariepinus to different levels of the pesticideunder daily renewal condition showed asignificant (p

JOURNAL OF ENVIRONMENTAL ENGINEERING AND TECHNOLOGY VOL. 1, NO. 2, MARCH 2012 22Table 1: Physio-Chemical Parameters of Exposure aquariaConc. Ofcypermethrin.(mlL -1 )pHConductivitys/cmDissolved(Oxygen)(mlL -1 )Alkalinity(mlL -1 )Hardness(mlL -1 )0.00 5.46±0.23 a 69.33±0.67 a 7.82±0.07 a 25.57±0.02 a 6.61±0.01 a0.05 6.07±0.27 a 69.75±0.75 a 7.50±0.30 a 24.48±0.28 a 6.07±0.03 a0.10 5.89±0.39 a 69.05±0.55 a 7.22±0.53 a 24.27±0.07 a 6.29±0.02 a0.15 5.87±0.45 a 70.15±0.55 a 7.08±0.42 a 24.16±0.41 a 6.04±0.01 a0.20 6.15±0.37 a 70.15±0.55 a 7.04±0.11 a 24.16±0.08 a 6.28±0.23 a0.25 6.06±0.41 a 69.80±0.56 a 6.85±0.56 a 24.77±0.58 a 6.36±0.05 aMean with the same superscript in the column are not significant different (P>0.05)22

JOURNAL OF ENVIRONMENTAL ENGINEERING AND TECHNOLOGY VOL. 1, NO. 2, MARCH 2012 23Table 2: Heamatological responses of Clarias gariepinus in various concentrations of solution of cypermethrin for 10 days (Mean±SD)Conc. Of cypermethrin0.00 0.05 0.10 0.15 0.20 0.25(mlL -1 )PCV (%) 34.76±3.06 a 30.00±6.25 ab 28.33±4.73 b 25.33±5.13 ab 15.66±10.02 ab 8.67±0.58 cHb (g/dl) 11.57±1.03 a 9.97±2.08 ab 9.47±1.57 b 8.43±1.96 ab 8.43±3.37 ab 3.97±0.25 cLeucocrit (%) 4.18±0.00 ab 3.33±2.31 a 2.00±1.00 ab 1.33±0.58 ab 1.00±0.00 b 1.33±0.587 abWBC (x 10 9 cells) 5.43±2.61 a 5.53±1.79 ab 6.20±0.69 ab 7.03±3.33 ab 11.23±0.06 b 15.23±1.29 aNeutrophils (%) 21.67±2.89 a 22.67±7.51 a 23.33±11.55 c 24.67±7.64 d 25.33±7.64 c 26.00±1.00 fLymphocytes (%) 78.33±20.21 a 71.33±20.13 a 71.67±15.14 a 71.67±18.93 a 71.67±15.28 a 41.33±8.08 abEosinophils (%) 10.00±17.32 a 8.33±14.43 a 5.81±4.00 b 4.67±8.08 b 3.41±5.00 b 3.33±8.62 aMonocytes (%) 4.33±5.77 a 3.33±14.43 a 3.01±0.66 a 2.67±3.06 a 1.67±2.89 a 1.01±0.10 aESR (mm/h) 2.33±0.58 b 2.01±1.53 b 3.97±1.15 b 5.67±2.52 b 7.87±2.08 b 14.67±6.43 aThrombocyte (%) 132.67±31.64 ab 120.00±0.58 ab 110.00±138.92 a 98.33±38.19 b 86.67±12.58 ab 53.33±40.72 bRBC 7.32±1.01a 6.01±1.11b 5.24±1.10c 4.01±1.02 ab 3.66±0.86 d 2.98±0.74MCHC (g/dl) 33.00±0.00 a 32.33±0.58 ab 31.67±0.58 c 30.00±0.00 abc 29.00±0.00 a 22.00±0.58 bcMCH (pg) 32.33±0.58 a 31.67±0.58 a 31.67±0.58 a 31.67±0.58 a 32.00±1.00 a 31.33±0.58 aMCV (fl) 96.67±0.58 a 96.33±0.58 a 95.33±1.15 ab 96.00±1.00 ab 96.33±2.08 a 93.33±2.31 bMean with the same superscript in the column are not significantly different (P

JOURNAL OF ENVIRONMENTAL ENGINEERING AND TECHNOLOGY VOL. 1, NO. 2, MARCH 2012 24IV. DISCUSSIONThe water quality variables in the experimentaltanks were within the range acceptable foroptimum performance of fish in culture medium(Boyd, 1998). There was concentrationdependent reduction in the values of dissolvedoxygen levels in the exposure tanks, an indicationthat cypermethrin affects the dissolved oxygenconcentration during the experimental procedureresulting in asphyxiation. This result agreed withthe findings of Gabriel et al. (2006) in Clariasgariepinus exposed to proxone in the laboratory.The haematological parameters in the controlgroup falls within the range of normal valuesreported for several cultured catfishes (Erondu etal., 1993: Ogbulie and Okpokwasili, 1999;Akinrotimi, 2008). Exposure of C. gariepinus tosublethal concentrations of cypermethrin caused asignificant decrease in the values of eosinophils,Hb, PCV, RBC, leucocrit, lymphocytes,monocytes and thrombocytes of the fish. Similarreductions were reported by some athors:Sampath et al. (1993) in Oreochromismossambicus exposed to OrganophosphorusOmoregie et al. (1994) in Oreochromis niloticusexposed to formalin, Svoboder et al. (2001) inCyprinus carpio exposed to diazinon and Gabrielet al. (2007) in C. gariepinus exposed to refinedcrude oil products kerosene. The reduction inRBC, Hb and PCV which are related to oxygencarrying capacity of the blood, may be due to theinhibition of erythropoiesis hemosynthesis, andincrease in the rate of erythrocyte destruction inhaemopoietic organs.Concentration dependent exposure of C.gariepinus to cypermethrin in this work caused areduction in the lymphocytes, leads tolymphopaenia, an indicator of unfavourableenvironment, thus corroborating the reports ofOluah and Mgbeka (2004) in C. albopunctatusexposed to sublethal concentrations of actellic25EC. Also, in this work, the reduction observedin leucocrit which is the leucocyte, thrombocytespacked value expressed as a percentage of thewhole blood is comparable to that of Adhikari etal. (2004) in fresh water teleost, Labeo rohitaexposed to cypermethrin and cabofuran underlaboratory conditions. This is an indication ofincipient stressful conditions of the fish.The reduction in the values of eosinophils andmonocytes reported in this study is in agreementwith the findings of Benariji and Rajendranath(1990) in Clarias batrachus exposed todichlorvos, but contrary to that of Velisek et al.(2007) in rainbow trout, after exposure todeltamethrin the disparity may be due to thedegree of potency of the toxicant and its mode ofactions in different species of fish. The ESRvalue of fish exposed to cypermethrin increasedsignificantly when compared to control values.This corroborates the report of Svobodova et al.(2003) in Cyprinus Carpio treated withdeltamethrin. The increase ESR value may bedue to fragility of erythrocytes as a result of stressinduces disruptions in the formation and releaseof erythrocytes from haematopaetic tissues(Akinrotimi et al., 2010b).Another type of haematological response to theeffect of cypermethrin was a significantincrement in WBC. This response was equallyobserved in common carp Cyprinus carpio afteracute effect of phenitrothion, imidan anddichlorvos (Svobodova, 1991, 1996, 1998). Thismay be due to release of white blood cells fromspleen into the blood stream to combat thetoxicant.Alteration in the erythrocytes indices of C.garipienus, consequent of cypermethrinexposure, caused a non significant change withno definite trend, this is in line with theobservation of Tsuda et al. (2007) in killifishsubjected to organophosphorus insecticide in thelaboratory.V. CONCLUSIONEvaluation of haematological parameters offish exposed to toxicants in this study, providedvaluable information in the assessment of fishhealth and in monitoring stress responses. Theexposure of C. gariepinus to sublethalconcentrations of cypermethrin caused markedvariations in the blood indices of the fish anindication that this chemical is highly toxic tofish and its use should be controlled so as toprevent its entrance into water bodies to minimizeaquatic pollution consequent of agrochemicals.REFERENCES[1] Adhikari, S., Sarkar B., Chatterjee, A., Mahapatra, C.T.and Ayyappan, S. Effects of cypermethrin andcarbofuran on certain haematological parameters andprediction of their recovery in a freshwater teleost Labeorohida. Ecotoxicology and Environmental Safety. 2004.58:220 – 226.[2] Akinrotimi, O.A. Comparative haematology of someculturable clariids raised in freshwater tidal and stagnantearthen ponds. M.Sc Thesis submitted to theDepartment of Fisheries, Rivers State University of24

JOURNAL OF ENVIRONMENTAL ENGINEERING AND TECHNOLOGY VOL. 1, NO. 2, MARCH 2012 25Science and Technology, Port Harcourt, Rivers State,Nigeria. 2008. 103pp.[3] Akinrotimi, O.A., Abu, O.M.G., Agokei, E.O. andUedeme-Naa, B. Effects of direct transfer to fresh wateron the haematological parameters of Tilapia guineensis(Bleeker 1862). Animal Research International 2010a.7(2):1199-1205.[4] Akinrotimi, O.A., Uedeme-Naa, B. and Agokei, E.O.Effects of acclimation on haematological parameters ofTilapia guineensis (Bleeker 1862). Science WorldJournal 2010b. 5(4):1-4.[5] Annune, P.A. and Ahuma, F.T.A. Haematologicalchanges in mudfish exposed to sublethal toconcentrations of copper and lead. Journal of AquaticSciences 1998. 13:33 – 36.[6] APHA American Public Health Association. StandardMethod for the Examination of Water and Waste Water.(17 th edition). Washington D.C, U.S.A. 1985. 1391pp.[7] Benarji, G. and Rajendranath, T. Haematologicalchanges induced by an organophosphorus insecticide ina freshwater fish Clarias batrachus. TropicalFreshwater Biology. 1990. 2:197-202.[8] Blaxhall, P.C. and Daisley, K.W. RoutineHaematological methods for use with fish blood Journalof Fish Biology. 1973. 5:771-781.[9] Boyd, C.E.. Water quality in warm water fish ponds.Aubun University Alabama. USA 1998. 122pp.[10] Bradbury, S.P. and Coast, J.R. Comparative toxicologyof the pyrethroid insecticides. EnvironmentalContamination Toxicology 1989. 108:134 – 177.[11] Erondu, E.S., Nnubia, C. and Nwadukwe, F.O.Haematological Studies on Four Catfish species raisedin freshwater ponds in Nigeria. Ichthyology 1993. 805:1-6.[12] Frank, R. Contamination of rural ponds with pesticides.Canadian Bulletin Contamination Toxicology. 1990.44:401-409.[13] Gabriel, U.U. Ezeri, G.N.O., Ikeme, P.M. and Ikwut,E.F. Haematoxicity of proxone to juvenile Clariasgariepinus (Burchell, 1822) under laboratory conditions.Pollution Research 2006. 25(2):1-6.[14] Gabriel, U.U., Amakiri, E.U. and Ezeri, G.N.O.Haematology and gill pathology of Clarias gariepinusexposed to refined petroleum oil under laboratoryconditions. Journal of Animal and Vetrinary Advances.2007. 6(3): 461-465.[15] Gagnon, A., Jumarie, C. and Hondela, A. Effects of Cuon plasma contisol and corrtisol secretion by adraanocoritical cells of rainbow trout. Aquatic Toxicology2006. 78:59-65.[16] McLeay, D.J. and Gordon, M.R. Leucocrit a simplehaematological technique for measuring acute stress insalmon fish, including stressful concentrations ofpulpmill effluent. Journal of Fishery Research 1977.34:2164 – 2175.[17] Meise, S.M., Siengupta, A., Lan, C., Zhou M andZhang, L. The common insecticides of cyfluthrin andchlorpyrifor alter the expression of a subset of geneswith diverse functions in fish. Toxicology Science. 2006.93:125-135.[18] Musa, S.O. and Omoregie, E. Haematological changesin Clarias gariepinus exposed to malachite green.Journal of Aquatic Sciences. 1999. 14:37-42.[19] Ogbulie, J.N. and Okpokwasili, G.C.. Haematologicaland histological responses of Clarias gariepinus andHeterobrancus bidorsalis to some bacterial diseases inRivers State, Nigeria. Journal Nature and Science. 1999.27(1):1-16.[20] Oluah, N.S. and Mgbenka, E.O. Effects of actellic 25ECon the leucocyte counts of the catfish, Clariasalbopunctatus. Animal Research International 2004.1(1):52-56.[21] Omoregie, E., Eseyin, T.G. and Ofojekwu P.C. Chroniceffects of formalin on erythrocyte counts and plasmaglucose of Nile tilapia Oreochromis niloticus. AsianFisheries science 1994. 7:1-6.[22] Perschbacther, P. and Ludang, G. Herbicide drift andChannel Catfish ponds. World Aquaculture 2002. 33(4):24 – 26.[23] Sampath, K., Velamnia, S.K. and James, R.Haematological changes and their recovery inOreochromis mossambicus as a function of exposureperiod and sublethal levels of Elalus. ActaHydriobiology 1993. 35:73-83.[24] Seiverd, C.E. Haematology for medical Technologists.(3 rd edition). Lea and Febiger, Philadelphia, U.S.A.1983. 946pp.[25] Svoboda, M., Luskova, V., Drastichova, J. and Zlabek,V. The effect of diazinon on haematological indices ofcommon carp Cyprinus carpio Acta Vetrinary Bruno.2001. 70:457-465.[26] Svobodova, Z., Luskova, V., Drastichovo, J., Svobovo,M and Zlabek, V. Effect of deltamethrin onhaematological indices of common carp Cyprinuscarpio. Acta Vetrinary Bruno 2003. 72:79-85.[27] Svobodova, Z., Machova, J., Kolarovo, J., Vykosovo, B.and Piacka, V. The effect of selected negative factorson haematological parameters of common carp Cyprinuscarpio. Review Zoology. 1996. 31:112 – 116.[28] Svobodova, Z., Pravda, D. and Palackova, J. Unifiedmethods of haematological examination of fish ResearchInstitute of Fish Culture and Hydrobiology, Vodnary,1991. 31pp.[29] Svobodova, Z., Vykosovo, B. and Groch, L. Pesticidesand fish poisoning. Agrochemicals and AnimalPoisoning. Acta Vetrinary Bruno. 1999. 60:58-62.[30] Tsuda, T., Kojima, M., Harada, H., Nakajima, A. andAoki, S. Acute toxicity, accumulation and excretion oforganophosphous insecticides and their oxidationproducts in killifish. Chemosphere 1997. 35:939-949.[31] Velisek, J., Juraikovo, J., Dosiskova, R., Svobodovo, Z.,Plackova, V., Machova, J. and Novotory, L.. Effects ofdeltamethrin on rainbow trout. EnvironmentalToxicology. 2007. 23:167-204.[32] Yaji, A.J. and Auta, J. Sublethal effects ofmonocrotophus on some haematological indices ofAfrican Catfish Clarias gariepinus. Journal of FisheriesInternational. 2007. 2(1):115-117.[33] Zar, J.H. Biostatistical Analysis. Prentice-Hall,Engelwood, Cliff, New Jersey, USA 1984. 260pp.25