Investigation Into Haematological Characteristics Of Parasite Infested Farmed African Catfish At Owerri North L.G.A, IMO State

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1 AGRICULTURE AND BIOLOGY JOURNAL OF NORTH AMERICA ISSN Print: , ISSN Online: , doi: /abjna , ScienceHuβ, Investigation Into Haematological Characteristics Of Parasite Infested Farmed African Catfish At Owerri North L.G.A, IMO State Agbabiaka, L.A 1 ; C.A. Ekeocha 1 and T.T. Akande 2 1. Department of Fisheries Technology, Federal Polytechnic, Nekede, Owerri 2. Department of Agricultural Technology,, Federal Polytechnic, Ado-Ekiti agbabiakala@gmail.com ABSTRACT Blood characteristics of parasites infested farmed catfish in Owerri-North Local Government Area, Imo State, Nigeria was investigated to evaluate the physiological disturbance(s) caused by parasitism. Forty African Catfish (Clarias gariepinus) were selected using a stratified random sampling technique in the study area. Sampled fish were tranquilized with 150mg/litre of tricane methane sulphonate (MS222) before being transported alive to the laboratory for heamatological analysis according to standard methods. Results indicated decrease values of Haemoglobin, Packed Cell Volume and Red Blood cells of sampled fish relative to the control group (uninfested fish) (p<0.05) while the White blood cells, lymphocytes and monocytes were least in the control (p<0.05). This study has shown that parasitic disease of fish has deleterious effect on the physiology and/or health status of the fish with potential economic loses to the fish farmers. Keywords: Blood, Catfish, infestation, Parasites, cultured INTRODUCTION Fish disease is an abnormal condition characterized by a gradual degeneration of a fish s ability to maintain normal physiologic function (Francis-Floyd, 2002). It is a condition in which the fish is not in balance with its self or its environment. Like any animal, fish are susceptible to range of problems such as tumours, heart and other organ disease, as well as metabolic disorders such as diabetes. However, the overwhelming majority of common health problems involve external parasites, fungi, viruses and bacterial infections. Most of the diseases might have occurred as a result of parasitic infestation or environmental pollution (Hussain et al., 2003). In Nigeria, the level of awareness of the impact of parasites and diseases to aquaculture is lacking as reported by Kolndadacha et al., (2007). Parasitic disease of fish is very common all over the world and is of particular importance in the tropic, fish being about the most parasitized of all vertebrate hence, the importance of parasitic infection on production has largely remained an issue of concern to fish farming industry (Roberts and Jonovy, 2000) In fish farming or aquaculture, some parasites may be highly pathogenic and contribute to high fish mortalities and economic loss, while in natural systems they may threaten the abundance and diversity of fish species (Mashego, 2001). Clarias gariepinusis one of the most resistant and widely accepted valued fish that is cultivated in Nigeria, therefore, the need for documented research on parasite which might constitute problems on this fish species especially in a cultured system cannot be over emphasized (Dankishiya and Zakari, 2007). Analysis of blood constituents is considered physiological indicator of the whole body and therefore, are important in diagnosis of the structural and functional status of fish exposed to pollutant (Agbabiaka et al., 2013a). In this respect, Ranzani- Paiva et al. (2000) demonstrated alteration in blood composition related to parasitism in fish from the Parana River, indicating that determination of blood parameters of fishes is of great importance in evaluation of disturbance that causes parasitism. Therefore, this study was aimed to investigate the impact of parasite on some blood parameters of Clarias gariepinus at Owerri North Local Government Area fish farms.

2 MATERIAL AND METHOD Study Area: The study was conducted at Owerri North Local Government Area, Imo State, Nigeria. Owerri North is semi-urban Local Government Area, and is located between the latitude 5 o 29ˈN and 7 o 2ˈN and longitude 7 o 28ˈE and 8 o 35ˈE Collection and transportation of fish specimen: Forty African Catfish (Clarias gariepinus) were selected randomly, four from each of the ten fish farms in the study area in a sterilized plastic containers labeled accordingly; fish were tranquilized with 150mg/litre of tricane methane sulphonate (MS222) according to Osuigwe et al. (2005) before being transported alive to the Fish Pathologist of Federal Polytechnic Nekede Owerri for blood collection and examination. Haematological evaluation: Blood samples were collected from each of the fish collected by severing the caudal peduncle using 2ml syringe and the blood samples poured into vial bottles containing Ethyl- Diamine Tetra-Acetic Acid (EDTA) after which it was mixed thoroughly to avoid clotting. Haematological indices: The haematological indices analyzed were as follows: Haemoglobin (HB) estimation was according to Dacie and Lewis (2000), Packed Cell Volume (PVC) according to Siwicki and Anderson (1993), Red Blood Cell (RBC), White Blood Cell (WBC), differential WBC count (Lymphocyte, Neutrophil and Monocyte counts) according to the method of Henry and Bergmeyer (1986), Mean Corpuscular Haemoglobin Concentration (MCHC) as described by Dacie and Lewis (2000), Mean Morpuscular Haemoglobin (MCH) according to Dacie and Lewis (2000), and Mean Cell Volume (MCV) determined according to the method of Dacie and Lewis (2000). Statistical analysis: Data obtained from the experiment were analyzed using the Analysis of Variance (ANOVA) and where significant differences occurred, means were separated using Duncan Multiple Range Test (Steel and Torie, 1980). RESULTS Table 1 shows the haematological characteristics of infested and (Control) uninfested catfish (Clarias gariepinus) that were randomly selected from different fish farms in Owerri North L.G.A. Haemoglobin (g/dl): The highest value of was recorded from the control group while the least value of 7.30 was obtained from farm E. There was significant difference between the control group (uninfested fish) and the infested fish (p < 0.05) Packed Cell Volume (%): The highest value of 42.60% was recorded from the control group while 21.00% was the least found in sample from farm E. There was significant difference between the control and the parasites infested sampled fish (p < 0.05). White Blood Cell (10 6 µl): The highest value of 5368 was recorded from farm E while the least value of 2233 was obtained from the control group. There was no significant difference between the sampled fish in farm A and C (p > 0.05) but significant differences occurred between the control and the parasites infested sampled fish in farm B, D and E ( p < 0.05). Red Blood Cell (10 6 /mm 3 ): The least value of RBC ( /mm 3 ) was recorded from farms B and E while the highest value of 3.30 x 10 6 /mm 3 was obtained from the control group. There was a significant difference between the control and parasites infested fish (p < 0.05). Lymphocyte (%): The Lymphocyte values ranges from 89.00% at farm A to 58.70% in the control group. There was significant difference between the control and the sampled fish (p < 0.05). Monocyte (%): The Monocyte value was highest in farm A, B, D and E group with the value of 4% respectively while the least value of 1.60 was recorded from the control group. There was no significant difference between the sampled fish in farm A, B, D and E (p > 0.05) but significant difference between control and the sampled fish in farm C (p < 0.05). Neutrophil (%): The Neutrophil value of was highest in control group while the least value of 7.00 was recorded in farm A. There was no significant difference between the sampled fish in farm B, C and E (p > 0.05) but significant difference between control and the sampled fish in farm A and D (p < 0.05) Mean corpuscular haemoglobin concentration (%): The highest value of was recorded in farm D while the least value of was obtained from the control group. There was a significant difference between the control and the sampled fish (p < 0.05). Mean Corpuscular Haemoglobin (pg): The highest value of was recorded from the control group while the least value of was obtained from farm E. There was no significant difference between the sampled fish in farms C and D (p > 0.05) but significant difference between control and the sampled fish in farms A, B, and E (p < 0.05) 188

3 Mean Cell Volume (fl): The highest value of was recorded in the control group while the least value of was obtained from farm E. There was no significant difference between the sampled fish in farms A and D (p > 0.05) but significant difference between in farms B, C and E (p < 0.05). Table 1: Haematological profile of parasite infested catfish (Clarias gariepinus) in Owerri Metropolis Farm Zones / Treatments Parameters 0 (control) A B C D E Hb (g/dl) a 8.30 b 7.70 b 8.50 b 8.60 b 7.30 bc PCV (%) a b b b b b WBC (10 6 µl) 2233 a 5308 b 5320 b 5056 bc 3264 c 5368 b RBC (10 6 /mm3) 3.30 b 3.10 a 3.07 a 3.09 a 3.15 a 3.07 a LYMP (%) a b b b c b MONO (%) 1.90 c 4.00 a 4.00 a 2.00 b 4.00 a 4.00 a NEUT(%) a 7.00 cd c c b 9.00 c MCHC(%) a b c b b c MCH (pg) a bc c b b c MCV(fl) a b bc b b c Means having the same superscript within a row are not statistically different (p > 0.05) Hb Haemoglobin P CV - Packed cell volume WBCs - White blood cell RBCs - Red blood cell MCV - Mean cell volume MCHC - mean corpuscular haemoglobin concentration MCH - mean corpuscular haemoglobin NEUT - Neutrophil LYMP - Lymphocytes MONO Monocytes DISCUSSION All the haematological parameters studied varied significantly (p < 0.05) between the control group (uninfested) and the parasites infested fish among the different farms in the study area. The haemoglobin values ranged from g/dl in the parasites infested fish samples while the control had the relatively higher value of 12.30g/dl. There was significant difference between the control and the parasite infested sampled fish (p < 0.05). The reduction in these haemoglobin concentrations relative to the control group may be due to the effect of parasitic disease which was observed previously by Klinger et al., (2002) and reported that the signs of nematodiasis include anemia, emaciation, unthriftness and reduced vitality. Normal Packed Cell Volume (PVC) ranged from 40-45% (Agbabiaka et al., 2013a), but this experiment showed that the packed cell volume (PVC) values were within 21-23% in the sampled fish with parasites while the control (non-parasitic fish) recorded the highest value of 42.60% (p < 0.05). The reduction of Parked Cell Volume in the sampled fish from the different farms may be as a result of the parasitic infestation that often leads to anemia in fish such as African catfish (Martin et al., 2004). More studies have revealed that parasites simply act as a stressor, and during primary stage of stress,the PVC changes due to release of catecholamine, which can mobilize the red blood cells from spleen (Wells and Webere, 1990) or it can induce the red blood cells to swell as a result of fluid shift into the intracellular compartment (Chocchia and Motals, 1989). Red Blood Cells (RBCs) of an organism determine the carrying capacity of dissolved oxygen and it is an estimation of the number of red blood cells per litre of blood. The results of the RBCs of this study showed values from x10 6 /mm 3 in the parasites invaded fish while the control had the highest value of 3.30 x10 6 /mm 3 (p < 0.05). These values are within the range reported by Dada and Ikuerowo (2009) when when ethanoic extracts of Garcinia kola seeds were fed to Clarias gariepinus broodstock but above the values recorded by Agbabiaka et al. (2013a), 189

4 Adeyemo (2007) and Gabriel et al. (2004) respectively. The increase in red blood cell concentration in the sampled fish may be as a result of low dissolved oxygen level which may indicate congenital heart disease and some lung disease in the fish which may be attributed to lapse in management of the farms such as poor feeding, water quality and overstocking that consequently may cause the pond to be polluted and decrease oxygen concentration inside the ponds. White Blood Cells (WBCs) are primary line of immunological defenses (Agbabiaka, 2012). The mean value of WBC of this study ranged from x10 6 µl in the sampled fish while the control had the lowest value of 2233 x10 6 µl. There was no significant difference (p > 0.05) between the sampled fish in farms A and C which may be that their management practice were similar but significant differences (p<0.05) occurred between the control and the sampled fish in farms B, D and E respectively. Changes in white blood cells may occur when fishes are stressed and environmental quality parameters are altered, Hussain et al. (2003) recorded increased white blood cells due to pathological response of parasitic infestation in the fish by stimulating the haemopoietic tissues and the immune system to produce antibodies and chemical substances as defense agent against infection or invasion by parasite. Lymphocytes are involved in the immune process, producing antibodies against foreign body such as parasites and responsible for a wide variety of immune system functions (Savari, 2011; Martins et al., 2004). The Lymphocyte values were between 82-89% in the sampled fish when compared with the control group that recorded 58.70%. There was significant difference between the control and the parasites infested fish (p < 0.05). These differences may be as a result of microbial infection or presence of foreign body or antigen in the circulatory system (Oyawoye and Ogunkunle, 1998). Monocytes protect the body of fish against bacteria and eat up small particles of foreign matters like parasites. In this study, the Monocyte values ranged from 2-4% in the sampled fish while the control had the lower value of 1.60%. There was no significant difference between the sampled fish in farm A, B, D and E (p > 0.05) which may attribute to their management practice but significant difference between control and the sampled fish in farm C (p < 0.05). The difference may be low immunity of the fish to fight against the parasite in the blood due to poor management practices at their respective fish farms. Neutrophil protect the body and against foreign bodies in the blood of the fish. In this study, the Neutrophil values are from 7-14% in the sampled fish while the control had the highest value of 16.40%. There was no significant difference (p > 0.05) between the sampled fish in farm B, C and E but significant difference occurred (p < 0.05) between control and the sampled fish in farms A and D. The reduction may be that the immunity of the fish are weak to fight against foreign organisms such as parasite in the blood cells which implies that the fish immune response and their resistance to pathogen may be affected by environmental temperature, stress, water contamination as reported by Ramy et al.( 2009). Mean Cell Volume (MCV) is an estimate of the volume of Red Blood Cells. However, the MCV values ranged from fl in the parasitized fish while the control had relatively higher value of 79.66fl (p > 0.05) but significant difference occurred between control and the sampled fish in farm B, C and E (p < 0.05). These values were below the observations of Agbabiaka et al. (2013b) when tigernut were fed to broiler finishers. The differences may be due to species difference or indicate presence of anemia as a result of nutritional deficiency in fish, which implies that fish may not have been fed with good quality feed rather they were often fed with obnoxious materials such as raw chicken intestine or dead chicken as observed during survey/sampling. Mean Corpuscular Haemoglobin (MCH) is the haemoglobin content of the average red cell. The MCH values are from pg in the sampled fish while the control had the lowest value of 66.14pg. There was no significant difference between the sampled fish in farms C and D (p > 0.05) but significant difference between control and the sampled fish in farms A, B, and E (p < 0.05). The reduction of Mean Corpuscular Haemoglobin in the sampled fish may be that the red blood cells are abnormally small and the blood of the fish lacks folic acid or vitamin B12 which causes anemia and it may be as a result parasitic infection and poor absorption of food etc. Mean Corpuscular Haemoglobin Concentration (MCHC) is the average Haemoglobin concentration in a given volume of Packed Red Cells. The MCHC ranged from 18 to 30g/dl and it is lower in fishes than 190

5 in mammals due to the space occupying nucleus (Bernardf et al., 2002). MCHC values ranged from % in the sampled fish while the control had the lowest value of 30.60%. There was a significant difference between the control and the sampled fish (p < 0.05). The increase in MCHC maybe as a result of parasitic infestation that often leads to anemia in fish such as African catfish (Martin et al., 2004); and also may be due to environmental stress in the farms. Statistical analysis obtain from the result showed significant difference (p<0.05) among the blood parameters of the control and the sampled fish within the study area. This may be attributed to the sex and management intensity of the farms, for example feeding quality. It was observed that most of the sampled fish were not fed with the complete/appropriate diets to boost their immunity in order to fight against diseases/ parasites; the stocking density often were beyond carrying capacity aiming to make more income from the farm, poor attitude of changing stale water in concrete ponds as may be required results in disease outbreak and also environmental stress. Farmers in the studied area are recommended to adopt Best Aquaculture practices that will guarantee improved ecosystem approach to fish farming and reduce attributes that may negatively influence the homeostasis of cultured African Catfish species that potentially predisposes fish to parasites and/or diseases. REFERENCES Adeyemo O.K (2007). Haematological profile of Clarias gariepinus (Burchell 1822) exposed to lead. Turkish Journal of Fisheries and Aquatic Sciences. 7: Agbabiaka LA (2012) Evaluation of Tigernut meal (Cyperus esculentus) as replacement for maize in African catfish and Broiler diets. Ph.D Thesis, Department of Animal Science and Fisheries, Imo state University, Owerri. Nigeria. 172p. Agbabiaka. L.A, F.N. Madubuike and B.U. Ekenyem ( 2013a). Haematology and serum characteristics of African catfish (Clarias gariepinus Burchell ) fed graded levels of tigernut based diets. American Journal of Experimental Agriculture. 3(4): Agbabiaka. L. A, F.N. Madubuike, B.U. Ekenyem and B.O Esonu (2013b). Effect of tigernut based diets on haematology and serum biochemistry of broiler finisher. Agriculture and Biology Journal of North America, 4(3): Bernardf, F., G. Joseph, Zinkl, C. Memi, and Jain, (2000). Schalms veterinary haematology. Lippincott and Wikins, Baltimore, Maryland. USA Chiooccchia, G., and R. Motals, (1989). Effect of catechokmines on deformability of red cells from trout. In: Relative roles of cyclic AMP and cell volume. Journal of Physiology, 412: Dacie, J. V. and S. W. Lewis, (2000). Practical haematological.9 th Edition. Churchhill Livingstone, London Dada A.A and Ikuerowo M (2009). Effect of ethanoic extracts of Garcinia kola seeds on growth and haematology of catfish (Clarias gariepinus) broodstock. African Journal of Agricultural Research. 4(4): Dankishiya, A. S. and M. Zakari, (2007): Study on the gastrointestinal helminth parasite of Clarias gariepinus (Tueyels). In Gwagwakda, FCT, Nigeria. BestJournal, 4(2): Frances-Floyd, R. (2002). Aeromonas infections. Florida Institute of Food and Agricultural Science, University of Florida, U.S.A. Gabriel U.U, Ezeri G.N.O and Opabunmi O.O.(2004). Influence of sex, source, health status and acclimation on the haematology of Clarias gariepinus (Bruchell 1822). African Journal of Biotech. 3(9): Hussain, S., Hassan, M. Z., Y. Mukhtar, and B. N. Saddiqui, (2003): Impact of environmental pollution in human behaviour and up-left of awareness level through mass media among the people of Faisalabad city. International Journal of Agriculture Biology, 5: Klinger, R. E. and R. F. Floyd, (2002). Introduction to Freshwater Fish Parasites. Florida Cooperative Extension Service; Institute of Food and Agricultural Sciences. University of Florida. http.//edis.ifasufl.edu. Accessed 20th June Kolndadacha, O. D., A. N. Okaeme, I. I. I. Ibiwioye, R. Y. Attribom, and Y. M. Musa, (2007): Fish disease control : A key success to aquaculture development in Nigeria. National institute for fresh water fisheries Research.New Bussa, Niger state. Nigeria Best Journal, 4(1): Martins, M. L., M. Tavares-Dais, R. Y. Fujimoto, E. M. Onaka and D. T. Nomura. (2004). Haematological alteration of Leporinus macrocephalus (Osteichtyes ; Anisakidae) in fish pond. Argentina Brasileiro Medical Veterinary. Zootecnia, 56: Mashego, S. N., (2001): Redistribution of Proteocephanlus graduligar. Annals of the Transvaal Museum, 38: Osuigwe D.I, Obiekezie A.I, Onuoha G.C. (2005). Some haematological changes in hybrid catfish (Heterobranchus longifilis x Clarias gariepinus) fed different dietary levels of raw and boiled 191

6 jackbean (Canavalia ensiformis) seed meal. African Journal of Biotechnology. 4(9): Oyawoye, E. O., and O Ogunkunle M.(1998). Physiological and biochemical effects of raw jack beans on broilers. Proceedings of Annual Comference of Nigeria Society for Animal Production, 23: Ranzani-paiva, M. J. T., Ishikawa, C. M., Das Eliras, A. A., and F elizardo, N. N. (2000). Haematological analysis of chara Pseudoplatystoma fasciatum in captivity. Aqua Responsible Aquaculture in the New Millennium. Ramy. R., Avtalion. L., and Williams, C. (2009).Environmental Control of the Immunoresponse in fish. Aquaculture, 11(2): Roberts, L. S., and Janovy, J.J. (2005). Foundation of Parasitology (seventh ed.). New-york, McGraw-Hill. Savari, A., Hedayati, A., A. Safahieh, and A. Movahedinia, (2011). Characteristics of blood cells and haematological parameters of yellow fin sea bream (Acanthopagruslatus) on some creeks of Persian Gulf. World Journal of Zoology, 6(1): Siwicki, A. K., and Anderson, D. P. (1993). Immune simulation in fish, and measuring the effects of stimulant by serological and immunological methods. Nordic symp. Fish immunology. Lysekil, Steel, R. G. D and J. H. Torie(1980). Principles and Procedure of Statistics. McGraw Hill Book. New York. Sudova, E., Piackova, V., Kroupova, H., Pijacek, M., Svobodova, Z. (2008) the effect of praziquantel applied per as on selected haematological and biochemical indices in Common carp (Cyprinus carpio L.). Fish Physiology and Biochemistry, 35(4): Wells, R. M. G., and R. E. Webere, (1990).The spleen in hypoxic and exercised rainbow trout. Journal Exp.Biol. 150:

Growth and Haematological Characteristics of African Catfish (Clarias Gariepinus Burchel, 1822) Fed African Star Apple Kernel Diets

International Journal of Zoology Studies ISSN: 2455-7269 www.zoologyjournals.com Volume 1; Issue 3; March 2016; Page No. 25-29 Growth and Haematological Characteristics of African Catfish (Clarias Gariepinus