The aim of this study was to assess the effects of a sublethal dose (1.6 mg/l) of DDVP (dichlor- vos) on hematological parameters of rainbow trout (Oncorhynchus mykiss) after 28 days of exposure. The DDVP caused increases in the red and white blood cell counts, hemoglobin, ery- throcyte sedimentation rate, mean corpuscular volume, and mean corpuscular hemoglobin con- centration (MCHC). On the other hand, it decreased thrombocyte (Plt), hematocrit (packed cell volume), and mean corpuscular volume. The only statistically significant differences between exposed and unexposed fish were in white blood cell count and MCHC

Aksakal, Ecrument

Angis, Simay

Atamanalp, Muhammed

Oguzhan, Pinar

eVols at University of Hawaii at Manoa

The Open Access Israeli Journal of Aquaculture – BamidgehAs from January 2010 The Israeli Journal of Aquaculture - Bamidgeh (IJA) will be published  exclusively  as  an  on-line  Open  Access  (OA) quarterly  accessible  by  all AquacultureHub  (http://www.aquaculturehub.org)  members  and  registered  individuals and institutions.  Please visit our website (http://siamb.org.il) for free registration form, further information and instructions.  This transformation from a subscription printed version to an on-line OA journal, aims at supporting the concept that scientific peer-reviewed publications should be made available to all, including those with limited resources. The OA IJA does not enforce author or subscription fees and will endeavor to obtain alternative sources of income to support this policy for as long as possible.Editor-in-ChiefDan Mires Editorial BoardSheenan Harpaz Agricultural Research OrganizationBeit Dagan, IsraelZvi Yaron Dept. of ZoologyTel Aviv UniversityTel Aviv, IsraelAngelo Colorni National Center for Mariculture, IOLREilat, IsraelRina Chakrabarti Aqua Research LabDept. of ZoologyUniversity of DelhiIngrid Lupatsch Swansea UniversitySingleton Park, Swansea, UKJaap van Rijn The Hebrew University Faculty of AgricultureIsraelSpencer Malecha Dept. of Human Nutrition, Food and Animal SciencesUniversity of HawaiiDaniel Golani The Hebrew University of JerusalemJerusalem, IsraelEmilio Tibaldi Udine UniversityUdine, ItalyCopy Editor Ellen Rosenberg Published under auspices ofThe Society of Israeli Aquaculture and Marine Biotechnology (SIAMB), University of Hawaii at Manoa Library andUniversity of Hawaii Aquaculture Program in association withAquacultureHub http://www.aquaculturehub.org              ISSN 0792 - 156X Israeli Journal of Aquaculture - BAMIGDEH.PUBLISHER: Israeli Journal of Aquaculture - BAMIGDEH -Kibbutz Ein Hamifratz, Mobile Post 25210, ISRAELPhone: + 972 52 3965809http://siamb.org.il The Israeli Journal of Aquaculture – Bamidgeh  60(1), 2008, 9-12. 9Alterations in Hematological Parameters of RainbowTrout (Oncorhynchus mykiss) Exposed to DDVPMuhammed Atamanalp*, Simay Angis, Pinar Oguzhan and Ercument AksakalFisheries Department, Agricultural Faculty, Ataturk University, 25240 Erzurum, Turkey(Received 12.6.07, Accepted 30.7.07)Key words: rainbow trout, DDVP, hematology, pesticideAbstractThe aim of this study was to assess the effects of a sublethal dose (1.6 mg/l) of DDVP (dichlor-vos) on hematological parameters of rainbow trout (Oncorhynchus mykiss) after 28 days ofexposure. The DDVP caused increases in the red and white blood cell counts, hemoglobin, ery-throcyte sedimentation rate, mean corpuscular volume, and mean corpuscular hemoglobin con-centration (MCHC). On the other hand, it decreased thrombocyte (Plt), hematocrit (packed cellvolume), and mean corpuscular volume. The only statistically significant differences betweenexposed and unexposed fish were in white blood cell count and MCHC. IntroductionDichlorvos (DDVP; 2,2-dichlorovinyl dimethylphosphate) is an organophosphate pesticidecommonly used to eradicate crustaceanectoparasites and treat sea lice (Lepophtheirussalmonis and Caligus elogatus) on commercialfish farms (Murison et al., 1997; Varo et al.,2003).Dichlorvos is volatile and highly toxic byinhalation, dermal absorption, and ingestion.As with all organophosphates, dichlorvos isreadily absorbed through the skin. Comparedto poisoning by other organophosphates,dichlorvos causes a more rapid onset ofsymptoms and is often followed by a similarlyrapid recovery (EXTOXNET, 1996). The toxi-city of DDVP has been studied in variousaquatic invertebrates and fish. Also, the dis-persion of the chemical after release from thecages has been simulated and monitored(Murison et al., 1997). Hematological parameters respond quick-ly to changes in environmental conditions andhave been studied in a wide range of marineto freshwater fish. Hematological parametersare used to describe fish health, monitorstress response, and predict systematic rela-tionships and physiological adaptations of ani-mals. Hematological parameters more quicklyreflect the poor condition of fish than othercommonly measured parameters.The goal of this research was to determinechanges in rainbow trout blood parametersfollowing exposure to DDVP. Materials and MethodsFish and maintenance. Rainbow trout of bothsexes (145±20 g) were obtained from theTrout Breeding and Research Center of theFaculty of Agriculture at Ataturk University.Seven fish were allocated to the treated group*  Corresponding author. E-mail: mataman@atauni.edu.tr10and five to the control. Fish were acclimated toconditions in the research unit for four weeksin 600-l fiberglass tanks with a fresh water flowof 0.5 l/min/kg fish, a waste water dischargeoutlet, and a water temperature of 11.0±0.5°C.Fish were fed commercial trout pelletsthroughout acclimation and the experiment. Toxicant. A commercial package of DDVPwas obtained from IMPA Tarim (Turkey). Alltanks were filled with 600 l water. Treatedtanks received DDVP at a sublethal concen-tration of 1.6 mg/l. The chemical was added tothe tanks once a day for 28 days. Blood collection and blood analyses.Approximately 2 cc blood was collected fromthe caudal vein of each fish, anticoagulantheparin was added, and the blood was placedin Vacutainer tubes. Red and white blood cellsand thrombocytes (Plt) were counted accord-ing to Blaxhall and Daisley (1973). The hemo-globin (Hb) concentration was determined asin Hoffmann and Lomel (1984), the erythro-cyte sedimentation rate as in Blaxhall andDaisley (1973), and the hematocrit (PCV) asin Schalm et al. (1975). The mean corpuscu-lar volume (MCV), mean corpuscular hemo-globin (MCH), and mean corpuscular hemo-globin concentration (MCHC) were calculatedaccording to Heath (1987) and as in Reddyand Bashamoiden (1989).Statistical analysis. Differences amonggroups were tested with variance analysesand group averages were analyzed usingDuncan’s test. ResultsThe red and white blood cell counts, Hb, ery-throcyte sedimentation rate, MCH, and MCHCwere higher in treated fish than in control fish(Table 1). On the other hand, thrombocyte(Plt), hematocrit (PCV), and MCV were lowerin treated fish. The only statistically significantdifferences were in white blood cell count andMCHC. DiscussionThe red blood cell count in the control groupwas within the range of Kocabatmaz andEkingen (1984; 0.538-1.185 x 106/mm3) andsimilar to that found by Atamanalp et al. (2002;0.603 x106/mm3). After DDVP exposure, thered blood cell count was higher in the treatedgroup but did not significantly differ from thecontrol. Likewise, this parameter increased inTilapia mossambica treated with cadmium(Aziz et al., 1993), Ctenopharyngodon idellaAtamanalp et al.Parameter Control (n = 5) Treated group (n = 7)Red blood cells (106/mm3) 0.64±0.19 0.65±0.17White blood cells (104/mm3) 6.9±1.84 11.79±1.76**Thrombocytes (104/mm3) 8.80±2.65 8.57±2.30Hemoglobin (g/100 ml) 6.90±2.46 7.70±2.21ESR (mm/h) 0.40±1.74 1.42±1.33Hematocrit (%) 43.00±12.40 41.00±11.16MCV (µm3) 661.54±212.84 655.75±200.84MCH (pg) 106.15±51.18 124.58±48.83*MCHC (%) 16.05±2.99 18.75±2.07ESR = erythrocyte sedimentation rateMCV = mean corpuscular volumeMCH = mean corpuscular hemoglobinMCHC = mean corpuscular hemoglobin concentration* = p<0.05; ** = p<0.01 Table 1. Hematological parameters of rainbow trout treated with dichlorvosfor 28 days, compared to untreated control fish (means±SD).treated with fenvalerate (Shakoori et al.(1996), Heteropneustes fossilis treated withdeltamethrin (Kumar et al., 1999), andOncorhynchus mykiss treated with cyperme-thrin (Atamanalp et al., 2002). On the otherhand, some researchers reported a decreasein red blood cell count after exposure, forexample, in Cyprinus carpio treated with fen-valerate (Reddy and Bashamohideen, 1989),Ctenopharyngodon idella treated with mercurychloride (Shakoori et al., 1991), and Anabastestudineus treated with monocrotophos(Santhakumar et al., 1999).The white blood cell count also increasedafter exposure to DDVP, similar to results ofShakoori et al. (1991), Aziz et al. (1993),Kumar et al. (1999), and Santhakumar et al.(1999). In contrast, Shakoori et al. (1996) andAtamanalp et al. (2002) reported that pesti-cides caused a reduction in white blood cellsin their studies. The number of thrombocytes (Plt) in thecontrol was much higher than obtained byKocabatmaz and Ekingen (1984) andAtamanalp et al. (2002) but did not statistical-ly differ from the value in the treated fish.Kocabatmaz and Ekingen (1984) reported 2.1x 104/mm3 thrombocytes in rainbow troutbefore stress and 4.3 x 104/mm3 after. Thesedifferences can be explained by differences instress level. Hemoglobin (Hb) was greater in treatedfish, perhaps because the oxygen carryingcapacity of the fish was affected by the DDVP.Dichlorvos appears to interfere with the abilityto bind hemoglobin to oxygen during respira-tion. Due to a possibly insufficient supply ofoxygen, respiration may have been affected.As a result, the demand for hemoglobinincreased. The increased hemoglobin mayalso be attributed to increased erythropoiesisand hemoglobin synthesis which, in turn,explains the increased MCHC. Sublethaldoses of mercury chloride increased thehemoglobin of C. idella (Shakoori et al., 1991)as did cadmium in T. mossambica (Aziz et al.,1993), fenvalerate in C. idella (Shakoori et al.,1996), and cypermethrin in O. mykiss(Atamanalp et al., 2002). In contrast, hemo-globin decreased in C. carpio treated with fen-valerate and cypermethrin (Reddy andBashamohideen, 1989), C. idella treated withfenpropathrin (Ahmad et al., 1995), andHeteropneustes fossilis treated withdeltamethrin (Kumar et al., 1999). DDVP caused a insignificant reduction inhematocrit concentrate (PCV), as it did in C.carpio exposed to fenvalerate (Reddy andBashamohideen, 1989), C. idella exposed tofenpropathrin (Ahmad et al., 1995), C. idellaexposed to fenvalerate (Shakoori et al.,1996), and O. mykiss exposed to cyperme-thrin (Atamanalp et al., 2002). The decreasein hematocrit concentrate suggests thatDDVP may interfere with the normal physiolo-gy of red blood cells. On the other hand, expo-sure to mercury chloride raised the hematocritconcentrate in C. idella (Shakoori et al., 1991)and T. mossambica (Aziz et al., 1993), possi-bly due to specific effects of this chemical in orreactions of these fish species. The erythrocyte sedimentation rate (ESR)of the control fish was similar to that of healthyrainbow trout found by McCarthy et al. (1975)but lower than that found by Kocabatmaz andEkingen (1984). ESR was greater in treatedfish, in agreement with Kumar et al. (1999),indicating that they were intoxicated by thetoxic chemical. ESR is raised by acute infec-tion, heavy metal toxicity, and kidney defor-mation (Blaxhall and Daisley, 1973). The difference in mean corpuscular vol-ume (MCV) between the control and treatedfish was not statistically significant. The dropin treated fish agrees with Shakoori et al.(1996) and Atamanalp et al. (2002). DDVP significantly increased the meancorpuscular hemoglobin (MCH) in the treatedfish. Toxicity and stress factors have beenreported to raise MCH (Reddy andBashamohideen, 1989; Aziz et al., 1993;Pages et al. 1995) or lower it (Shakoori et al.,1996; Kumar et al., 1999). This discrepancymay originate from the chemicals or differ-ences among fish species. The mean corpuscular hemoglobin con-centration (MCHC) rose in treated fish, similarto Shakoori et al. (1991, 1996) but in contrastto Reddy and Bashamohideen (1989), Aziz etal. (1993), and Santhakumar et al. (1999). 11Hematological parameters of rainbow trout exposed to DDVP12This research shows that dichlorvos hasprofound effects on the hematological para-meters of rainbow trout. More such studiesare needed because different life stages with-in this species and other species may reactdifferently.AcknowledgementsThe authors thank Dr. Nurinisa Esenbuga forthe statistical analyses and Dr. E. MahmutKocaman for the fish. References Ahmad F., Ali S.S. and A.R. Shakoori, 1995.Sublethal effects of Danitol (fenpropathrin), asynthetic pyrethroid, on freshwater Chinesegrass carp, Ctenopharyngodon idella. Folia.Biol. (Krakow). 43:151-159.Atamanalp M., Yanik T., Haliloglu H.I. andM.S. Aras, 2002. Alterations in the hemato-logical parameters of rainbow trout,Oncorhynchus mykiss, exposed to cyperme-thrin. Israeli J. Aquac. - Bamidgeh, 54(3):99-103.Aziz F., Amin M. and A.R. Shakoori, 1993.Toxic effects of cadmium chloride on thehaematology of fish, Tilapia mossambica.Proc. Pakistan Cong. Zool., 13:141-154.Blaxhall P.C., 1972. The haematologicalassessment of the health of freshwater fish: Areview of selected literature. J. Fish. Biol.,4:593-604.Blaxhall P.C. and K.W. Daisley, 1973.Routine haematological methods for use withfish blood. J. Fish Biol., 5:771-781.EXTOXNET, 1996. Pesticide, InformationProfiles, Extension Toxicology Network.http://extoxnet.orst.edu/pips/dichlorv.htm.Heath A.G., 1987. Water Pollution and FishPhysiology. CRC Press. p. 245.Hoffmann R. and R. Lomel, 1984. Effects ofrepeated blood sampling on some blood para-meters in freshwater fish. J. Fish Biol.,24:245-251.Kocabatmaz M. and G. Ekingen, 1984. Thestandardization of blood sampling and hema-tological methods in different species of fish.Doga Bilim Dergisi D1, 8(2):149-159 (inTurkish with English summary).Kumar S., Lata S. and K. Gopal, 1999.Deltamethrin induced physiological changesin freshwater catfish Heteropneustes fossilis.Bull. Environ. Contam. Toxicol., 62:254-258.McCarthy D.H., Stevenson J.P. and M.S.Roberts, 1975. Some blood parameters ofthe rainbow trout (Salmo gairdneriiRichardson). II. The Sasha variety. J. FishBiol., 7:215-219.Murison D.J., Moore D.C., McHenery J.G.,Robertson N.A. and I.M. Davies, 1997.Epiphytic invertebrate assemblages anddichlorvos usage at salmon farms.Aquaculture, 159:53-66.Pages T., Gomez E., Suner O., Viscor C.and L. Tort, 1995. Effects of daily manage-ment stress on haematology and blood rheol-ogy of the gilthead sea bream. J. Fish. Biol.,46:775-786.Reddy P.M. and M.D. Bashamoiden, 1989.Fenvalerate and cypermethrin inducedchanges in the haematological parameters ofCyprinus carpio. Acta. Hydrochim. Hydrobiol.,17(1):101-107.Santhakumar M., Balaji M. and K. Ramudu,1999. Effect of sublethal concentrations ofmonocrotophos on erythropoietic activity andcertain hematological parameters of fishAnabas testudineus. Bull. Environ. Contam.Toxicol., 63:379-384.Schalm O.W., Jain N.C. and E.J. Carrol,1975. Veterinary Haematology, 3rd ed. Lea &Febinger, Philadelphia.Shakoori A.R., Iqbal M.J., Mughal A.L. andS.S. Ali, 1991. Drastic biochemical changesfollowing 48 hours of exposure of Chinesegrass carp Ctenopharyngodon idella, to sub-lethal doses of mercuric chloride. Proc. 1stSymp. Fish Fish., Pakistan. pp. 81-98.Shakoori A.R., Mughal A.L. and M.J. Iqbal,1996. Effects of sublethal doses of fenvaler-ate (a synthetic pyrethroid) administered con-tinuously for four weeks on the blood, liverand muscles of a freshwater fish,Ctenopharyngodon idella. Bull. Environ.Contam. Toxicol,. 57:487-494.Varo I., Navarro J.C., Amat F. and L.Guilhermino, 2003. Effect of dichlorvos oncholinesterase activity of the European seabass (Dicentrarchus labrax). PesticideBiochem. Physiol., 75:61-72.Atamanalp et al.

Alterations in Hematological Parameters of Rainbow Trout (Oncorhynchus mykiss) Exposed to DDVP

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Alterations in Hematological Parameters of Rainbow Trout (Oncorhynchus mykiss) Exposed to DDVP

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