Sie sind auf Seite 1von 12

Continental J. Pharmacology and Toxicology Research 4 (1): 18 - 29, 2011 ISSN: 2141 4238 Wilolud Journals, 2011 http://www.wiloludjournal.

l.com ` Printed in Nigeria

HAEMATOLOGICAL RESPONSES OF Clarias gariepinus EXPOSED TO ANAESTHETICS METOMIDATE Gabriel, U.U., 2Deelae. S.N. 3Akinrotimi, O.A 4Orokotan, O.O. Department of Fisheries and Aquatic Environment, Faculty of Agriculture, P.M.B. 5080, Rivers State University of Science and Technology, Port Harcourt, Rivers State, 3African Regional Aquaculture Centre/Nigeria Institute for Oceanography and Marine Research P.M.B. 5122, Port Harcourt, Rivers State, 4 Gabrovic Agric Nig. Ltd. Fishery Consultancy Service 100 East West Road, Eliozu Junction, Rumuodara, Port Harcourt, Rivers State, Nigeria. Corresponding author:Akinrotimi , E-mail:ojoakinrotimi@yahoo.com
1,2 1

ABSTRACT Haematological responses of Clarias gariepinus exposed to different levels of anaesthetic metomidate were investigated, using varying concentrations (0.00 control, 0.25, 0.75, 1.00, 2.00, 4.00, 6.00, 8.00, 10.00 and 12.00 mL-1) in triplicates. The result obtained from the study indicates that the anaesthetic caused a concentration dependent changes in the blood of C. gariepinus with significant reduction (P < 0.05) in the values of Haemoglobin (Hb), Packed Cell Volume (PCV), Leucocrit (Lct), White Blood Cell (WBC), Platelets (PLT), Monocytes (Mon) and red blood cell (RBC), while the values of Erythrocyte sedimentation Rate (ESR) and Eosinophils (EOS) increased significantly (p < 0.05) with concentration, which was more pronounced in the fish exposed to higher concentrations of 10.00 and 12.10mL-1 INTRODUCTION The use of anti-stressors is a common practice in modern aquaculture. Such substances are used to induce anaesthesia during handling and sorting, tagging, and artificial reproduction procedures, thus reducing stressinduced problems such as decrease in feeding and immune functions (Ross and Ross 1999). The anaesthetics most commonly used in aquaculture are MS-222, benzocaine, quinaldine sulphate, metomidate, clove oil and 2phenoxy ethanol (Svoboda and Kolarova, 1999: Waterstrat, 1999; Velisek et al., 2007). Anaesthesia is usually induced by immersing the fish in an anaesthetic solution. For appropriate application of these chemicals, Guilderhus and Marking (1987) defined three criteria that an anesthetic applied in aquaculture most fulfil,l effectiveness, wide safety margin and inexpensive. The criterion of efficacy of any anesthetics depends on a number of factors, including the concentration of an anesthetic, water temperature, fish size and species (Mcfarland, 1960; Feireira et al., 1984; Soto and Burhanddin, 1995). The efficacy of 2-phenoxyethenol, MSS222 - TMS and metomidate have been documented for many species: Sockeye salmon (Sehdeve et al., 1963); Rainbow trout (Guilderhus and Marking, 1987; Iwama et al., 1988), Platy fish (Guo et al., 1992); grass and silver cap (McCarter, 1992); Storgeon hybrid (Di Marco, et al., 2011); gold fish (Kaiser and Vine, 1998); yellow perch (Hamackova et al., 2001); sea bream (Tort et al; 2002) tench (Myszkowski et al., 2003; Hamachova et al., 2004) common carp (Velisek and Svobodova, 2004a). However, there are no such reports on any of the clariids. Yoshikawa et al. (1994), noted that unanesthesized fishes suffer a great stress during catching, transportation or when subjected to various aquaculture manipulations, (such as relocation from) its natural environment and artificial propagation. The stress and pains can distort on the physiological responses, (haematological and biochemical profiles) and inducing endocrine and metabolic disturbances (Yokohoma et al., 1993). Hence, anesthesia of the fish could minimize biochemical and physiological disturbances due to stress. However, the anaesthetic drug itself can induce undesirable effects also distorting biochemical and haematolgoical results. Investigations revealed that tricaine methanesulphonate (MS-222), the most commonly used drug, leads to physiological and biochemical disturbances, if the recommended doses is exceeded (Morales et al., 1990; Velisek et al., 2007). Nonetheless, Hurin and Greer, (1991) reported no major changes in blood chemistry after anesthesia with tricaine and benzocaine. Until recently, tricaine methanesuphonate (TMS, MS-222) is the only anaesthetic common in Nigeria, there are now many brands used in the market for aquacultural purposes. One of such drugs is metomidate. Metomidate, marketed as a fish anaesthetic under the trade name Transquil , is a rapid-acting non barbiturate hypnotic with

18

Gabriel, U.U et al.,: Continental J. Pharmacology and Toxicology Research 4 (1): 18 - 29, 2011

potential cortisol suppressing properties. Metomidate is an effective anaesthetic in both fresh and salt water and has been reported to be more potent in adult salmon adapted to sea water (Olsen et al., 1995). In assessment of the negative effects of this analysis of blood indices have proved to be reliable indices especially for aquatic organisms (Velisek et al., 2009). The aim of this study therefore was to investigate the haematological responses of C. gariepinus a common aquaculture species in Nigeria, exposed to anaesthetic metomidate, at various concentrations. MATERIALS AND METHODS Anaesthetic Metomidate hydrochloride with the trade name tranquil manufactured by Syndell Aqua Ltd, Canada, was purchase off shelf from Gabrovic Agric Ltd, Rumuodara, Port Harcourt in Rivers State, Nigeria. Experimental Procedure Thirty three gravid female fish ready to spawn, brood fish (mean length 60.23cm0.36SD and 2,640kg 1.36SD) were obtained from a private fish farm (Water Shed Farm, Rumuodara, Port Harcourt, Rivers State, Nigeria), and transferred into fishery laboratory in Department of Fishery, Rivers State University of Science and Technology, Port Harcourt, Rivers State, Nigeria. They were acclimated to laboratory conditions for a period of 10 days. After this, they were exposed to solution of metomidate hydrochloride in different concentrations (0.00-control, 0.25, 0.50, 0.75, 1.00, 2.00, 4.00, 6.00, 8.00, 10.00 and 12.00 mlL-1) in 30L aquaria in triplicates for a period of 1 hour, when, the fish in all the exposure concentrations have become immobilized (Velisek et al., 2009). Blood Sampling and Analysis Blood was collected from the caudal vessel of the fish with a 21 Gauge hypodermic needle and 5 ml disposable syringe, and transferred immediately into blood sample bottles containing EDTA (ethylene-diamine tetra-acetic acid) which is used in routine haematological investigation in human medicine and gives good result with preserved blood. After collection of blood samples, they were analyzed in laboratory using Standard methods as described by Akinrotimi et al. (2010). Red Blood cell RBC) were counted immediately, in haemacytometer (Improved Neubauer, Weber Scientific Limited, Middlesex, UK) using the method of Wintrobe (1934). Packed Cell Volume (PCV) was determined by filling herparinised haematocrit capillary tubes with blood and centrifuged for 5 minutes at 500xg in a micro capillary reader (Serveid, 1983). Haemoglobin was done using cyanmethemoglobin methods (Blaxhall and Daisley, 1973). The total white blood cell counts (WBC) were similarly enumerated in an improved haemocytometer using shaws diluting fluid (Miale, 1982). Blood smear were stained, using Grumwald-Giensa stain, for differential examination (Tavares Dias et al., 1999). Statistical Analysis One way analysis of variance (ANOVA) was used to test for the significance (P < 0.05) of the effects of anaesthetic on fish blood. The differences among were analyzed by ANOVA using the SPSS statistics package (SPSS 9.0, SPSS, Chicago, IL, USA), and multiple comparisons were performed using Turkeys comparison test (Wahua, 1999). RESULTS Generally, treatment with the anesthetic caused a progressive, decrease (p < 0.05) in the values of PCV, Hb, Leucocrit, WBC, Platelets, monocytes and RBC, (Fig 1, 2, 3, 5, 6, 10 and 11). But reverse was the case with ESR, neutrophils, eosinophils and MCV (Fig. 4, 7, 9 and 14) while the responses of lymphocytes, MCHC and MCH relative to metomidate concentrations varied with no definite pattern (Figs 8, 12, 13). C. gariepinus treated with metomidate had highest PCV (35.067.86%), Hb (13.060.64g/dl) and leucocrit (4.007.13%) value at the control and lowest at 12.00ml/L (Fig. 1, 2). ESR (11.413.22mm/hr) was recorded at exposed 12.00ml/L concentration and the lowest value (2.141.21mm/hr), observed in the fish under control fish (Figs. 4).

19

Gabriel, U.U et al.,: Continental J. Pharmacology and Toxicology Research 4 (1): 18 - 29, 2011

The highest values of WBC (22.642.71mm/hr) and platelets (90.763.86mm/hr) were recorded in the fish exposed to 0.00ml/L concentration and the lowest values were recorded in the 12.00ml/L concentrations (Figs. 5 and 6). The highest values of neutrophils were observed in the fish exposed to 12.00ml/l concentration, while the lowest value was in the control trial (fig. 7). The highest value of lymphocytes (66.646.12mm/hr) was observed in the fish exposed to 0.00ml/L, while the lowest value in 0.50ml/L (fig 8). The lowest value of eosinophils was recorded in the fish exposed to 1.00ml/L concentration and those values tends to increase as the concentration levels increases and peaked at 12.00ml/L. (Fig. 9). The value of Monocytes recorded during the trial has no definite trend, the lower values were observed from 6.00 to 12.00ml/L level of metomidate concentration (Fig. 10). The red blood cell (RBC) values decreased as the concentration level increases with the highest and lowest values recorded in the control and 12.00ml/L respectively (Fig. 11). The red blood indices (MCH, MCV, MCHC) recorded during this trial has no particular tends in relation to the level of concentrations (Figs 12, 13 and 14).

PCV(%)

Concentration (m/L )

-1

Fig. 1: Changes in PCV of C. gariepinus exposed to metomidate anaesthetics at different concentrations. Different letters on the bars indicate statistical significance (p<0.05)

Haemoglobin (g/dl)

Concentration (m/L )

-1

Fig. 2: Changes in Haemoglobin of C. gariepinus exposed to metomidate anaesthetics at different concentrations. Different letters on the bars indicate statistical significance (p<0.05)

20

Gabriel, U.U et al.,: Continental J. Pharmacology and Toxicology Research 4 (1): 18 - 29, 2011

Leucocrit (Cell x 1012/L)

Concentration (mg/l)
Fig. 3: Changes in Leucocrit of C. gariepinus exposed to metomidate anaesthetics at different concentrations. Different letters on the bars indicate statistical significance (p<0.05)

ESR (mm/hr)

Concentration (mg/l)
Fig. 4: Changes in ESR in C. gariepinus exposed to metomidate anaesthetics at different concentrations. Different letters on the bars indicate statistical significance (p<0.05)

WBC (Cells x 10 /L)

Concentration (mg/l)
Fig. 5: Changes in WBC in C. gariepinus exposed to metomidate anaesthetics at different concentrations. Different letters on the bars indicate statistical significance (p<0.05)

21

Gabriel, U.U et al.,: Continental J. Pharmacology and Toxicology Research 4 (1): 18 - 29, 2011

Platelets (Cells x 109/L)

Concentration (mg/L-1) Fig. 6: Changes in Platelets of C. gariepinus exposed to metomidate anaesthetics at different concentrations. Different letters on the bars indicate statistical significance (p<0.05)
Neutrophils (%)

Concentration (mg/L-1) Fig. 7: Changes in Neutrophils of C. gariepinus exposed to metomidate anaesthetics at different concentrations. Different letters on the bars indicate statistical significance (p<0.05)

22

Gabriel, U.U et al.,: Continental J. Pharmacology and Toxicology Research 4 (1): 18 - 29, 2011

Lymphocytes (%)

Concentration (mg/L )

-1

Fig. 8: Changes in Lymphocytes of C. gariepinus exposed to metomidate anaesthetics at different concentrations. Different letters on the bars indicate statistical significance (p<0.05)

Eosinophils (%)
Concentration (mg/L) Fig. 9: Changes in Eosinophilis of C. gariepinus exposed to metomidate anaesthetics at different concentrations. Different letters on the bars indicate statistical significance (p<0.05)

Monocytes (%)

Concentration (mg/L ) Fig. 10: Changes in Monocytes of C. gariepinus exposed to metomidate anaesthetics at different concentrations. Different letters on the bars indicate statistical significance (p<0.05)

-1

23

Gabriel, U.U et al.,: Continental J. Pharmacology and Toxicology Research 4 (1): 18 - 29, 2011

RBC (Cells x 106/L)

Concentration (mg/L) Fig. 11: Changes in Red Blood Cell of C. gariepinus exposed to metomidate anaesthetics at different concentrations. Different letters on the bars indicate statistical significance (p<0.05)

MCHC (%)

Concentration (mg/L) Fig. 12: Changes in MCHC of C. gariepinus exposed to metomidate anaesthetics at different concentrations. Different letters on the bars indicate statistical significance (p<0.05)

24

Gabriel, U.U et al.,: Continental J. Pharmacology and Toxicology Research 4 (1): 18 - 29, 2011

MCH (pg)

Concentration (mg/L) Fig. 13: Changes in MCH of C. gariepinus exposed to metomidate anaesthetics at different concentrations. Different letters on the bars indicate statistical significance (p<0.05)

MCV (FI)

Concentration (mg/L) Fig. 14: Changes in MCV of C. gariepinus exposed to metomidate anaesthetics at different concentrations. Different letters on the bars indicate statistical significance (p<0.05)
DISCUSSION The effects of anaesthetics on haematological characteristics is an important tool to monitor various types of stressors that can infer the fish population health, as haematological indices are closely related to response of the animal to the environment. According to Gomes et al. (2003), haematological response in fishes differed depending on the nature, type and duration of the stressor. In evaluating the effects of anesthetics, MS222 on haematology of giant cods, Dissothichu mawsoni and Pagothenia borchgre,it was discovered that these fishes have a reduction in the haemoglobin and heamatocrit and blood pH (Wells et al., 1984). In sea water adapted mummichogs, Fundulus heteroclitus, anaesthesis with MS-222 and metomidate resulted in swollen and appreciably elevated blood lactate (Velisell et al., 2005), while Abdolazizi, et al. (2011) reported tubular necrosis and decreased of haematocrit in Carassius auratus exposed to clove oil. Weber et al. (2009) confirmed that the introduction of clove oil, 2-phenoxyethanol, and metomidate anaesthetics at high levels in water that contain silver perch (Perca flaviatillis) did in fact affect haematological parameters and enzymes profiles of the brood fish, and hatchability of eggs. Also, Velisek et al. (2003), in common carp Cyprinus carpio, showed that exposure of this fish to varying degrees of anaesthetics caused a decrease in

25

Gabriel, U.U et al.,: Continental J. Pharmacology and Toxicology Research 4 (1): 18 - 29, 2011

haemoglobin, haematrocrit and dysfunctional enzyme activities. Furthermore, they observed that the fish exposed to metomidate for 96hrs showed a decline in blood glucose, haemoglobin, haematocrit, and produce a negative trend in blood immunological functions. The exposure of C. gariepinus to various levels of metomidate, led to a decrease in the values of packed cell volume (PCV) and haemoglobin (Hb) similar to a finding by Myszkowski et al. (2003) in tech Tinca tinca exposed to the anesthetic 2-phenoxyethanol. The decrease in the values of PCV and Hb could be attributed to haemolysis resulting in haemodilution, a mechanism for diluting the concentration of the anaesthetics in the circulatory system of the fish (Smith, et al., 1979). Changes in the values of WBC in brood C. gariepinus was similar to the one reported by Sudagal et al. (2009) in poach, (Rutilus rutilus) on application of 2-phenoxy ethanol. The decrease in the number of WBC (leucopoenia) may be as a result of the bioconcentration of anesthetics in the liver and kidneys (Agrawal and Srivastava, 1980). The bio-concentration appeared to be concentration-dependent as reflected in the values of WBC. The reduction observed in leucocrit, platelets and lymphocytes may be due to the reaction of fish to the effect of stress induced by anaesthetics. This was attributed to a generalized stress response resulting from increased pituitary interrenal activity (Thompson and Eling, 1989). Holloway et al. (2004) reported a significant reduction in lymphocytes and platelets of rainbow trout, after exposure to clove oil and MS-222. The concentration dependent increase (p > 0.05) in the values of neutrophils, eosinophils, and monocytes under metomidate agrees with the report of Wey et al. (1996), in gold fish, Carassius auratus exposed to 2-phenoxy ethanol at different temperatures and concentrations. The increase in number of these differential counts may be due to recruitment of more cells to resist the stressor. CONCLUSION In summary, metomidate anaesthetics was associated with changes in blood parameters of C. gariepinus. However, the effects of the anaesthetics were different from one another in the measured variables. On the basis of this experiment, it appears that metomidate effects on the blood parameters is concentration dependent, with the highest effects observed in the fish exposed to 12.0Ml/L. Hence application of metomidate should not be too high, not exceeding 8.0m/L-1 in application. REFERENCES Abdolazizi, S. Ghaderi, E., Naghdi, N. and Barzan B.K. (2011). Effects of Clove oil as an anaesthetic on some haematological parameters of Carassius auratus. Aquaculture Research and Development 2(1):1-3. Ackerman, P. A. (2006). Stress hormones and the cellular stress response in salmonids. Journal Fish Physiology Biochemistry. 23:236-240. Agrawal, S. J. and Srivastava, A. K. (1980). Haematological responses in a freshwater fish exposed to experimental manganese poisoning. Toxiocology 17:97-100. Ainsworth, A. J; C. Dexlag and P. R. Waterstrat (1991). Changes in peripheral blood leukocyte percentages and function of new cells in stressed channel catfish. J. Aquat. Anim. Health 3:41-47. Akinrotimi, O. A., Gabriel, U. U., Anyanwu, P. E. and Anyanwu, A. O. (2007). Influence of sex, acclimation methods andperiod on haematology of Sarotherodon melanotheron. Res. J. Biol. Sci. 2(3): 348-352. Akinrotimi, O.A., Abu, O.M.G., Uedeme-Naa, B. and Aranyo, A.A. (2010). Haematological characteristics of Tilapia guineensis from Buguma Creek, Niger Delta, Nigeria. Electronic Journal of Environmental, Agricultural and Food Chemistry 9(8):1415 1422. Di Marco, P., Petochi, T., Longoband, A., Priori, A., Finoia, M.G. Donadelhi, V., Corsalini, I, and Marino, G. (2011). Efficacy of tricaine methane sulphonate, clove and metomidate Ketamine and their side effects on the physiology of sturgeon hybrid, Acipenser nacarii X Acipenser baerii. J. Appl. Ichthyol. 27:611 617. Fereira, J. T; Shoonbee, H. J. and Smith G. L. (1984). The uptake of the anaesthetic benzocaine hydrochloride by the gills and skin of three freshwater fish species. Journal of fish Biology (25):35-41.

26

Gabriel, U.U et al.,: Continental J. Pharmacology and Toxicology Research 4 (1): 18 - 29, 2011

Gomes, L. C; Duran E; Gasquez, S; Mosotz,M and Roncero, V. (2003). Effects of fish density during transportation on stress and mortality of juvenile tambaqui Colossoma macropomum. J. World Aquacult. Soc. 34(1):76-84. Graham, M. S. and Iwama, G. K. (1990). The physiologic effect of the anaesthetic, ketamine hydrochloride on two salmons species. Aquaculture, 90:323-332. Guilderhus P. A. and Marking L. L. (1987). Comparative efficacy of 17 anaesthetic chemicals on rainbow trout. North American Journal of Fish management (7):288-292. Guo F. C; Teo L. H and Chea T. W. (1992). Effects of anaesthetics on plasma cortisol and lactic and levels in Platys (Xiphophorus maculates). Bulletin of Faculty of Science Natural University, Singapore (12):30-33. Hamackova, J; Sedpva, M. A; Pjaova, S. V. and Lepicova, A. (2001). The effect of 2-phenoxyethanol, clove oil and propiscin anesthetics on perch (Perca fluviatilis) in relation to water temperature. Czech J. Anim. Sci. 46:469-473. Hamockova, J; Lepicova, A; Kozakp; Stupla, Z. Kouril, J and Lepic P. (2004). The efficacy of various anaesthetics in tench (Tinca tinca) related to water temperature Vet. Med. 49:467-472. Holloway A. C., Keene J; Noakes, D. G; Moccia, R. D. (2004). Effects of clove oil and MS-222 on blood hormones profiles in rainbow trout Oncorhyrchus mykiss. Aquacult. Res. 35:1025-1030. Iwama G. K; McGeer, J. C. and Pawluk, M. (1988). The effect of five fish anesthetics on acid-base balance, haematrocirit, blood gasses, cortisol and adrenaline in rainbow trait. Can J. Zool. 67:2065-2073. Kaise, H and Vine, N. (1998). The effect of 2-phenoxyethanol and transport packing density on the porttransport survival rate and metabolic activity in the goldfish, Carassius auratus. Aquarium Science and Conservation (2):1-7. Kawauchi, H. (1989). The duality of teleost gonado tropins. Fish Physiol Biochem. 7:29-38. Laidley C. W., Leatherland, J. F and Moryen J. F. (1988). Cohort sampling, anaesthesia and stocking density effects on plasma cortisol, thyroid hormone, metabolite and ion levels in rainbow trout, Salmo gairdneri; J. Fish, Biol. 33:73-88. Levavi-sivan, B. and Avita, A. (2005). Sequence analysis, endocrine regulation and signal transduction of reactive cholesterol pools in mitochandria isolated from gonads of male goldfish (Carassius auratus). Gen. Comp. Endocrinol. 142:67-73. Levavi-Sivan, B; Avitan, A; and Kanias, T. (2003). Characterization of the inhibitory dopamine receptor from the pitutary of tilapia. Fish Physiol. Biochem. 28:73-75. McCarter, N. (1992): Sedation of grass carp and silver carp with 2-phenoxyethanol, clove oil and propiscin anaesthetics on perch (Perca fluviatilis) in relation to water temperature. Czech Journal of Animal Science (46):469-473. McFarland W. M. (1960): The use of anesthetics for the handling and transport of flounder. California Fish Bull. (46):406-431. Miale, J.B. (1982). Laboratory Medicine haematology (6th edition). The CV Mushy Publishers, London. 863pp. Morales, A. E., Garcia R and Higuera M. (1990). Influence of handling and/or anaesthesia on stress response in rainbow trout: effects of liver primary metabolism. Comp. Biochem. Physiol. 95A:87-94. Myszkowski L; Kaminski R. and Wolnick J. (2003). Response of juvenile tench Tinca tinca to the anaesthetic 2phenoxyethanol. J. Appl. Ichtch. 19:142-143.

27

Gabriel, U.U et al.,: Continental J. Pharmacology and Toxicology Research 4 (1): 18 - 29, 2011

Ross, L. G. and Ross, B. (1999). Anaesthetics and sedative technique for Aquatic Animals. Blackwell Science, Oxfrod, UK, 76pp. Seiverd, C.E. (1983). Haematology for medical technologists. Lea and Fabiger, Philadelphia, USA 590pp. Smith, G. L., J. Hattingh and A. P. Burger (1979). Haematological assessment of anesthesia, MS-222 in natural and neutral forming three freshwater fish species: interspecific differences. J. Fish Biol. 15:633-643. Snieszko, S. F. and Axerfold, R. H. (1989). Disease of Fishes. T. H. publication Inc., New Jersey 30pp. Sudagar, M., Mohammaeizarajabae, A., Mazandarani, R. and Pooralimottengh, S. (2009). The efficacy of clove powder as an anaesthetic and its effects on haematological parameter on roach (Rutilus rutilus) Journal of Aquaculture Feed Science and Nutrition. 1:1-5. Svobodova M. and Kolarova J. (1999). A survey of anesthetics used in the fish farming (in Czech) Pp. 49-72. In Health Protection of Fish-Proceeding of papers, Research Institution of Fish Culture and Hydrobiology, Vodonany, Czech Republic. Tavares Dias, M., Moraes, F.R. and Pandi C.L. (1999). Physiological responses of tambaqui Colossoma macropomom to acute stress. Bulletin do Instituto de pesca Sao Paulo 27(1): 43 48. Thompson, D. and Eling, T. (1989). Mechanism of inhibition of prostaglandin H. synthase by eugenol and other phenolic perioxidase substrates. Mol. Phamacol. 36:809-817. Tort, L; Puigcerver, M., Crespo, S. and Padros F. (2002). Cortisol and haematological response in sea bream and trout subjected to the anesthetics clove oil and 2-phenoethanol. Aquaculture Research 33:907-910. Velisek J. and Svobodova Z. (2004a). Anesthesia of rainbow trout (Oncorhynchos mykiss) with 2phenoxyethanol: acute toxicity and effects on biochemical blood profile. Act Vet. Brno. 73:378-384. . Velisek J., and Svobodova. Z. (2004): Anesthesia of rainbow trout, Oncorhynchus mykiss. Acta Veterinaria Brno, (74): 139-146. Velisek, J. and Svobodova, Z., Piackova V., Groch L., Nepekchalovah G.H(2003). Effects of clove oil anesthesia in comm. carp. Vet. Med. 50:269-274. Velisek, J., Wlasow T., Gomulka, P, Z., Svobodova and L. Novotry (2007). Effects of 2-phenoxyethanol anesthesia on sheatfish Silvurus glanis. Vet Med. 52(3):103-110. Velisek, J., Stesjskal, V., Kouril, J. and Svobodovo, Z (2009). Comparison of the effects of four anaesthetic on Biochemical blood profiles of Perch. Aqualculture Research 40:354 361. Wagner, E., Arudt. R. and Hilton, B. (2002). Physiological stress responses, egg survival and sperm mobility for rainbow trout brood stock anaesthetized with clove oil, tricaine methane sulfonate and carbon dioxide. Aquaculture 211:353-366. Wahua, T. A. T. (1999). Applied Statistics for scientific studies. Afrika link books. Aba, Nigeria. 365pp. Waterstrat P. R. (1999): Induction and recovery from anesthesia in channel catfish Ictalurus punctatus fingerlings exposed to clove oil. Journal of the World Aquaculture Society. (30): 250-255. Weber, R.A., Peleteiro, J.B., Garcia, Martin, L.O. and Aldegunde, M. (2009). The efficacy of 2-phenoxyethanol, metomidate, clove oil and MS-222 as anaesthetic agents in the Senegalese sole (Solea senegalensis) Aquaculture 288:147-130. Wedemeyer, G.A and Yasutake, W. T (1977). Clinical Methods for the Assessment of the Effect of environmental Stress on Fish Health. US Fish and Wild Life services, Washington. Tech Paper 89, 19pp.

28

Gabriel, U.U et al.,: Continental J. Pharmacology and Toxicology Research 4 (1): 18 - 29, 2011

Wells, R. M. G., Tetens, V. and DeVries, A. L. (1984). Recovery from stress following capture and anesthesia of Antarctic fish haematology and blood chemistry. J. Fish Biol. 25:567-576. Wey, O., Kaises, H and Hetch, T. (1996). On efficacy and mode of action of 2-phenonoxy ethanol as an anesthetic for gold fish, Carasius auratus at different temperatures and concentrating, Aquaculture Research 27:757-764. Wintrobe, M.M (1934). Variations of the size and haemoglobin content of erythrocytes in the blood of various vertebrates. Folia, Haematology Leipzig 51:32 49. Yokohama, Y; Kawai F. and Kanamori, M. (1993). Effect of cold CO2 anesthesia on post-mortem level or ATP related compounds, pH and glycogen in carp muscle. Nippon Sunsan. Gakkabhi 59:2047-2052. Yoshikawa, H; Kawai F., and Kanamori, M. (1994). The relationship between the EG and brain pH in the carp, Cyprinus carpio subjected to environmental hypercapnia at an anaesthetic level. Comp. Biochem. Phyisiol, 107A: 307-312. Received for Publication: 26/06/11 Accepted for Publication: 26/08/11 Akinrotimi, O.A African Regional Aquaculture Centre/Nigeria Institute for Oceanography and Marine Research P.M.B. 5122, Port Harcourt, Rivers State, Nigeria. E-mail:ojoakinrotimi@yahoo.com

29