International Journal of Scientific & Engineering Research, Volume 4, Issue 5, May ISSN

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1 International Journal of Scientific & Engineering Research, Volume 4, Issue 5, May Helminthology and Haematological parameters of S. labiatus, in Shallabugh Wetland &River Sindh Ibraq Khurshid and Fayaz Ahmad Post Graduate Department of Zoology University of Kashmir, Srinagar , Kashmir Abstract Helminthology and Haematology parameters have been recognised as the important tools for assessing the fish health. Haematological parameters were studied and compared different feeding behaviour of fishes in the two water bodies. Blood parameters such as red blood cell count (RBC) and white blood cells count (WBC), haemoglobin and ESR were estimated from fishes of different trophic level. Statistical analysis revealed that differences in haematological parameters between lotic and lentic fish were significant (P<0.01). The result revealed that haematological RBC/WBC ratio and ESR were significantly correlated at P<0.05 level. The RBC level decreases due to the increase in the helminth infection. WBC and ESR increases due to increase of helminth infection. Key Words: Helminth, Schizothorax, River Sindh, Shallabugh Wetland 1. INTRODUCTION For the last few decades, fishes have been extensively used as a protein rich diet for human consumption in India and thus, contribute a lot to its economy. According to Tatcher (1981) many parasites can live in a host, sometimes causing damage, sometimes not. Therefore, the changes associated with haematological parameters due to various parasites establish a data base and allow precise diagnosis guiding the implementations of treatment or preventive measures which are indispensable in fish farming and fish industry (Roberts, 1981). Knowledge of the haematological characteristics is an important tool that can be used as an effective and sensitive index to monitor physiological and pathological changes in fishes. Normal ranges for various blood parameters in fish have been established by different investigators in fish physiology and pathology (Rambhaskar and Srinivasa Rao 1986; Xiaoyun et al. 2009). The analysis of blood indices has proven to be a valuable approach for analysing the health status of fish as these indices provide reliable information on metabolic disorders, deficiencies and chronic stress status before they are present in a clinical setting (Bahmani et al. 2001). Therefore, the present study was designed to study the haematological abnormalities of fish fauna of Shallabugh wetland 2013

2 International Journal of Scientific & Engineering Research, Volume 4, Issue 5, May and River Sindh, Kashmir arising due to helminth infections so that necessary steps are taken to improve the health condition of these economically important fish fauna. 2. Hypothesis It was predicted that with increase in the helminth infection haematological parameters vary and with increase in length the prevalence of infection increases. 3. Materials and Methods This study was conducted on 179 live fish specimens belonging to family Schizothoracinae. Live fish specimens were collected from different sites of Shallabugh wetland and River Sindh,which were then taken to the laboratory in large water containers. The fish were identified by using the key provided by Sven et al. (1999). The fish were acclimatized to standard laboratory conditions for 12 hrs and were subjected to haematological and helminth parasitic investigations. For haematological investigations, blood samples were collected from all fish hosts in glass tubes containing EDTA and were properly labeled. For estimation of haemoglobin content (Hb), routine method was employed. The RBC and WBC counting methods were based on the dilution of obtained blood with dilution fluids (Hayem and Turk) in RBC and WBC counting pipettes (Mitruka and Rawnsley, 1977). Individual cells were then counted in the counting chamber (Haemocytometer). Giemsa s staining method was used for the differential count of WBC. The parasitic worms were collected according to the routine parasitological techniques and proper record was maintained. 4. Study site: Shallabugh wetland is 16 km northwest of Srinagar and measures about 12 square kilometres while the River Sindh contains the fresh flowing melting water of the glaciers.the Wetland consists of larger area of riverine marshes and shallow fresh water with associated reedbeds on the floodplains of river Jhelum. The Wetland together with marshes is fed by the Anchar Lake and local runoff. The average depth of the water varies from 0.3 to 2.0 meters. The Wetland supports a locally important fishery and reed harvesting Industry, which provides excellent opportunities for sport Hunting. 5. Map of study area 2013

3 International Journal of Scientific & Engineering Research, Volume 4, Issue 5, May Experimental Design Three collection sites were selected for collection in Shallabugh wetland. One site is where the Anchar Lake meets the Shallabugh Wetland (inlet), second is the middle of Shallabugh Wetland and the third is the sangam (outlet). In River sindh wayil, wahidpora and Shadipora was selected as the collection sites. 7. Data Analysis Pearsons correlation was used to asses association between different physic chemical features and helminth infection. Chisquare analysis was also used for calculating the prevalence of helminth infection and the relation between length and infection rate. 8. Results and discussion 2013

4 International Journal of Scientific & Engineering Research, Volume 4, Issue 5, May Prevalence, Mean intensity in relation to length of S. labiatus Factors like age, weight, length and sex of the host have been found to play a significant role in the Helminth infection of fishes. In the present study length of fish was correlated with the infection parameters like prevalence and mean intensity. In this present table it is clearly mentioned that with increase in length, parasitic infection goes on increasing. Table 1. showing Prevalence, Mean intensity in relation to length of S. labiatus Host Location Length No. No. No. of Prevalence Mean examined positive parasites Intensity S. Shallabugh labiatus Wetland 5.5 River Sindh S. SW labiatus 20.5 RS S. SW labiatus 25.5 RS S. SW labiatus 30.5 RS Total Table 2. Relationship of host length and parasites distribution of S. labiatus in Shallabugh Wetland Host Mean length No infected No. uninfected S. labiatus S. labiatus S. labiatus S. labiatus DF = 6, P-Value = Table 3. Correlation of host length and parasitic burden of S. labiatus in Shallabugh Wetland Host Mean length No. of parasites S. labiatus 13 5 S. labiatus S. labiatus S. labiatus DF = 3, P-Value = Correlation of length and No. of parasites =

5 International Journal of Scientific & Engineering Research, Volume 4, Issue 5, May Table 4. Relationship of host length and parasitic distribution of S. labiatus in River Sindh Host Mean length No infected No. uninfected S. labiatus S. labiatus S. labiatus S. labiatus DF = 6, P-Value = Table 5. Correlation of host length and parasitic burden of S. labiatus in River Sindh Host Mean length No. of parasites S. labiatus 13 1 S. labiatus 18 6 S. labiatus S. labiatus DF = 6, P-Value = Correlation of length and No. of parasites = From above tables our findings demonstrate that there exist significant positive relationship between total length and the number of worms per fish. So we can say that the prevalence of intestinal helminth infection in fish species examined increases with length. The probable reason for differences in prevalence of infection between the juvenile and the adult fish as related to their length may be due to change in diet from weeds, seeds, Phytoplanktons and Zooplanktons to insect larvae, snails, crustaceans, worms and fish in both juveniles and adulthood respectively (Read et al., 1967). Previous studies have also reported the positive relationsship between level of infection and size of the host fish ( Dogial 1970 and Arai 1998)., Similarly Positive relationship between parasitic burden and host size in S. niger can be due to the fact that larger fish provides more surface area for the parasite attachment (Nie and Kennedy 1991; Koskivaara 1992). Anosike et al.1992 reported that number of parasites and its diversity increase with age of fish. Geets and Ollevier 1996 fish19 see choti dairy reported that increase in the abundance of parasites with host size. Brouder 1999 stated that Round tail chubs, G. robusta infected with Asian tapeworm showed a significant difference in total length compared to uninfected chubs, and a positive correlation between infection intensity and total length of fish (Brouder 1999). Kouyn and altunnel 2007 reported a positive significant correlation between the fish length and infection rate. Tekin-Ozan et al concluded from their results that the prevalence of parasites increased according to length of fish host. The increase in the infection with the increase in length may be due to the ageing factor of the fish, Oniye and Aken Ova , Mohammed et al.2009 reported prevalence was found to increase as the fish grows, and that could be attributed to the longer time of expose to the environment by body size. 2013

6 International Journal of Scientific & Engineering Research, Volume 4, Issue 5, May Haematology and blood biochemistry picture of uninfected and infected Fish hosts. Haemato- biochemical parameters have been recognized as valuable tools for monitoring fish health. Haematological and serum biochemical parameters were studied and compared different feeding behaviour of fishes. The blood from the hosts was screened for the haematological parameters including Hb. Definite variations between the fish inhabiting the different water bodies were detected in all haematological and biochemical values. However, significant differences in the haemato-biochemical were observed between values of the groups (infected and uninfected) in this study, suggesting that in the intensive helminth infection elevated haematological and biochemical parameters will be observed in the fish. Table 6. Haemotological picture of uninfected and infected S. labiatus Shallabugh wetland River Sindh p-value Haematological parameters Uninfected (mean) Infected (mean) Uninfected (mean) Infected (mean) Hb 8.40± ± ± ±0.42 T=19,P= RBC 2.86± ± ± ±0.34 T=12.63, P=0.05 TLC 1.19± ± ± ±0.28 T=10.82,P=0.05 ESR 1.35± ± ± ±0.62 T=13,P=0.04 The Hb thus shows ve correlation with the prevalence of infection and decreases as the intensity of infection increases.the same results were obtained by Ivasik and Virepo (1969) in carp due to sanguinicolosis in which haemoglobin reduced by 20% in mild infection and 61% in serious cases; Evans (1974) in cut throat trout due to Sanguisicola kiamathensis, Skvortosva (1977) in carp infected with Dilepis unilateralis larvae; Natrajan and Balakrishnan (1977) in Hemirhamphus xanthopterus infected with Lerncteenicus hemirhamiphi, Kawatsu (1978) in crucian caip infected by Diplozoon nipponicum, The Hb thus shows ve correlation with the prevalence of infection and decreases as the intensity of infection increases.the same results were observed by; Voznyi et al (1975 ) for carp with parasitic infections; Natrajan et al (1977) in Hemirhamphns xanthopterus and H. far infected with Lernaeenicus hemirhamphi; Denisov (1979) in silver carp infected with Posthodiplostomun jiicokt wherein RBC count decreased by 25%; The total leucocyte count increased in all fish species depending upon the intensity of infection to combat against the pathogenicity of the parasite Denisov (1979) in silver carp infected with Posihodiplostonntm cuticola due to which TLC rose by 44%, Saxena el al. (1993) in Heteropneiisteus fossils infected with Lucknoma indica due to which TLC increased by 3.72% 2013

7 International Journal of Scientific & Engineering Research, Volume 4, Issue 5, May The ESR thus showed positive correlation with the prevalence of infection and increased with increase in infection intensity. The same results were obtained by Voznyi et.al (1975) for carp with parasitic infections, Denisov (1979) for silver carp infected with Posthodiplostomum cuticola where in ESR increased 2 to 4 fold and Agarwal et.al ( 1989 ) for fresh water fish, Rita rita infected with trematodes, thus supporting our results. REFERENCES Agarwal (1989). Studied on haematology of trematode infected fresh water fish Rita rita (Ham.). Ind. J. Helminthol. 41: Bahmani M, Kazemi R, Donskaya P (2001) A comparative study of some hematological features in young reared sturgeons (Acipenser persicus and Huso huso). Fish Physiol Biochem 24: Denisov A.K. (1979). The pathogenic effect of Posthodiplostomum cuticola infection on silver carp. Bolezni rybi bor ba snimi. 23: Dubey N.K. (1980). Studies on some Pathological aspects of certain fresh water Air- Breathing Fishes.Ph.D. Thesis. LNMU, Darbhanga. Engelherdt, A., Le Mirle and H. Peterman, Hematological studies into rainbow trout affected by Proteocephalus neglectus. Monatsh Veterin Aermed. 44: Evans W.A. (1974). Growth, mortality and hematology of cut throat trout experimentally infected with the blood fluke Sanguinicola Kiama thensis. J. Wild Life Dis. 10: Mushiake K., Mori K. and Arimoto M. (2001). Epizootiology of anaemia in wild Japanse flounder, GYOBYO Kenkyu-Fish Pathol. 36: Roberts R.J. (1981). Patologia de los Peces. Madrid, Mundi-Prensa, pp: 366. Rambhaskar B, Srinivasa Rao K (1986) Comparative haematology of ten species of marine fish from Visakhapatnam Coast. J Fish Biol 30: Ivasik V.M. and Virepo B.G.S. (1969). Sanguinicolosis in carp in the winter, Helminthologia (Bratisl). 10: Mitruka B.M. and Rawnsley H.M. (1977). Clinical, biochemical and haematological reference values in normal experimental animals. Mason Publishing, NewYork, USA. Sarwar S.G. (1999). Water quality and Periphytic algal component of Anchar lake, Kashmir. In: In land water resources, India. In: Durga Prasad, M.K. and P. Sankara Pitchaich (Eds.). Discovery publishing house, New Delhi). pp:

8 International Journal of Scientific & Engineering Research, Volume 4, Issue 5, May Satpute L.R. and Agrawal S.M. (1974). Parasitic effects on its haematology and Histopathology. Ind. J. Exp. Biol. 12: Saxena K.K. and Chauhan R.R.S. (1993). Effects of parasitic infection on the blood of Heteropneustus fossilis (Bloch). Bioved. 4: Shamim A.B. and Pandit A.K. (2002). Limnological investigation of Anchar Lake, Kashmir. J. Res. Dev. 2: Shukla G.S. and Upadhyay (1998). A text book of economic zoology. Rastogi Publications. investigations on pigeons and clarias batrachus carrying cestode infections. Ann. Zool. 10: Sinha K.P. (2000). Hematological manifestation in clarias batrachus carrying helminthic infection. J. Parasitic Dis. 24: Sven O. Kullander, Fang Fang, Deping B. and Erik Ahlander (1999). The fishes of the Kashmir Valley pp In: Nyman L. (Ed.). River Jhelum, Kashmir Valley-impacts on the aquatic environment published by Swedmar. The international Board of Fisheries, Sweden. Tatcher V.E. (1981). Patologia de peixes da Amazonia brasileira, 1. Aspectos gerais. Acta Amazonica. 11: Yoshinaga T., T. Kamaishi I. Segawa K. Yamano, H. Ikeda and M. Sorimachi (2001). Anaemiacaused by challenges with the monogenean Neo heterobothrium hirame in Japanese flounder. Gyobyo Kenkyu-Ish Pathol. 36: Yousuf A.R. and Shah G.M. (1988). Comparative limnology of some fresh water habitats of Kashmir. Geobioas New Reports. 7: Zutshi (1980). Comparative limnology of nine lakes of Jammu and Kashmir Himalayas. Hydrobiol. 72:101. Xiaoyun Z, Mingyun L, Khalid A, Weinmin W (2009) Comparative of haematology and serum biochemistry of cultured and wild Dojo loach Misgurnus anguillicadatus. Fish Physiol Biochem 35:

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