Je Env, Zool, Vol. 4, 1990, pp, 166-172 EFFECTS OF SUBLETHAL TOXICITY OF SEVIN (CARBARYL) ON BLOOD PARAMETERS IN CLARIAS BAIRACHUS (L). (Received October 15, 1990; Accepted December 20, 1990) Birendra Kumar & V. Banerjee Haematology Laboratory, Department of Zoolozy, Patna University, Patna—809 005, India Changes in blood parameters of Clarias barrachus due to exposure to sub-lethal concentration of Sevin (17.1 mg/l) for 15,30, 45 and 60 days are reported. There was rapid fluctuations in blood parameters upto 45 days, Cell surface and nuclear surface area increased but CEG, Hb level and PCV decreased sign icantly even after 60 days of exposure. Sevin (Carbaryl), a methyl carbamate is extensively used all over the world to control agricultural as well as non-agricultural pests. It has been reported to cause adverse effects on reproductive physiology in several animal species. In long term studies, disturbances of carbohydrate and protein metabolism and protein metabolism and endocrine function has been reported. It has been selected for this study because it is frequently used by Indian farmers. However, a few investigations of pesticide toxicity at sublethal concentration on fish blood have appeared’. As the authors found no similar record with ‘Sevin, therefure this report is on the effects of sublethal exposure of Sevin on the blood parameters of @ common Indian siluroid air-breathing fish Clarias batrachus (L). MATERIALS AND METHODS Adult healthy Glarias batractus collected from local catchers were acclimatized inEffects of Sublethal Toxicity of Sevin on Blood [ 107 ine boratory in large glass aquaria (1.5% 1,0%0.5 m) undor natural photoporiod Im temperature (21°C —23°C) during January to February 1989. Fishes ranging In weight from 16 to 20 cm were considered for the study. During acclimation fishes Wore fed with minced goat liver which was stopped before 24h of the experiment In each case. Preliminary experiments were performed according to rocommonded procadur The 48h LCyy value for this fish was 51.6 mg/l and 1/3 of this was taken as gublethal concentration, Ten fish were kept in each four 2001 glass aquarium Containing top wator and 10 fich wore kept in anathar four such aquaria containing sublethal concentration of Sevin. There was no fish mortality during the entire course uf the experimont and the water in contral and water with pesticide in expo ced fish aquaria were renewed after 90h to keep the pesticide concentration at requisite level. After 15, 30, 45 and 60 days, 10 fish of one control and 10 treated fish of one aquarium were sampled for blood collection by cutting the caudal pedun- cle region by sharp sterilized razor blade". For morphological and differential counts blood film from each fish was prepared, atained in haemotoxylin and eosin. For cell dimensions evar micrometer (1 dive3.65 um) was Used. The Total Erythrocyte Count (TEC). fb content (Hb). Packed Gel! Volume (PVC). Mean Corpuscular Haemoglobin (MCH), Mean Corpuscular Fenes globin Concentration (MCHG), Mean Call Volume (MCV), Erythrocyte Sedimes teem mate. (ESR) and Total Leucocyte Count (TLC) wore determined after earlier she significant difference was recorded by applying student‘ test at RESULTS Variatians in blood parameters due to sub-lethal exposure of Sovin in Clarias atrachus nave been included in Table 1. Thara was fluctuation of the blood parame tare considered in this study but the effect was woll marked only after 60 ays. ‘There was significant increase in length of erythrocyte nucieus, eryihrocyle wud nuclear surface areas, tendancy of cell and nuclous {0 ovalness, significant decrease in TEC, Hb level, PCV and MCH. There was also significant increase in ESR and inKumar & Sanerie® rerers of Jigs bahavhus exposed Co sublethal Changes in Blood param seyte and its nucleus; Band Secin, Land L=tength of evytheacyte and ae ma ite mucleas; GA and NAorythrocyte aud nuclear a te; M=rmonocyte; N-neutro. Himensions 1000 erythrocytes and ed were considered, Table 1 ebreadeh of erythro sorface area: LL and SLo-large and small lye inophil |; For erythrocyte d phil: Ecosinophil; Bebasophil; For ery for other values 10 tadividuals of cach control and treat On 18th di On soth day On 45th day On Goth day ro Treated Control Traated Control Treated Control Treated Parameter Control Treated Loferythrocyte (am) ¥.S3 187 IZ HO ioe 9.35 10.88 B of erythrocyte (am) 8.81 7.76 8.68 ae 9.85 U of nucleus (#m) 20 3.56 25 3.64" 5! of nucleus (+m) 3.08 sat 06 3.29 us 17d 1168 L061 L103 us 1.056 L146 1.062 1,107 CA (am?) 55.65 69.15 64,75 84,25 NA (x4) 1.64 a7 78st OAL NACA 0.136 0.125 0,120 0.111 TEC (Xtot/mm' 2.85 219 2851.97" Hg (9%) 11,60 9.68 8.90 6,78" PCV (%) 21.98 20.19 21,03 17,70° MCV (em?) 94.53 95.55 95.44 92.03 92,72 89.54 MCH (pg) 51.86 45.39 1546 4442 37,80 34.28" MCHC (9%) 54.68 48.51 49.26 4748 47.93 40,77 38.28 ESR (mm/h) 2.02 274 3A7 W297 ag 1.90 Total and Difterontial leucocyte Counts (%) St oa 7. 35 7759.9 778 gs LL 159 Ie 20,395 14.0 Ae wee 1eL8 M 108 7.7 12.7 8.0 0 69 ne uD N as 59 72.758 86 42 aA 57 E 30 1.9 OG 23 Lo me 3.8 B 19 09 La 04 12, 0.3 Ll 0.6 TLE (K10%mm*) 7S 12.30 9.74 "9.561099 ag 6g os *Significant at P<0.05Effects of Sublethal Toxicity of Sevin on Blood [ 169 TLC along with small lymphocyte percentage and decre ase in all other types (Table 1), ot leucocyte DISCUSSION In the blood film preparations of treated fish there were erythrocytes of different shapes and hypochromasia was evident but the authors found no cells with vacuoles or dividing cells as were earlier raported in other fishes due to pes! cide treatments",%°. The significant increase in erythrocyte and nuclear surface ‘areas towards the end of the experiment showed the long term effect of Sevin on the erythrocytes of thie fish. Exposure in sublethal concentration of Sevin to Clarias butrchus resulted in gradual decrease in TEC, Hb and PCV with increased period of exposure. This study vas continued for G0 days and so the values of each blood parameter were deter vrined for contro! and treated fish separately to avoid soasonal variations®. TEC, Hb and PCV showed fluctuation in tho beginning but decreased significantly after 60 days. The absolute values showed irregular decrease or increase but decreased at the and of the experiments Thus the Sevin acted like thiment 10G, elsan 2% dust bazanon 5G (Channa punctatus), dichlorvos",* (a sslis and Clariar balrachie), Lindane®™ (Hy fost) Dut differs {rom the works of other authors'** who found Increase in TEC, Hb and PCV values due to sublethal exposure of pesticides. Inthe present work lowering of TEC indicated low activity of haemopoietic organs of the fish due to pesticide which continued till 60 days showing interferen CMlved forming elements by Sevin for a long period which may continue for days fagether until te fIsii become abnormal or dia. The long term toxic effect of Sevin ig beyond doubt ac the values did not return to wormal values after 60 days. in Savin treated fislh the omall lymphacytes increased sionificantly and all other types of leucocytes recorded significant decrease,Kumar & Baneriee 170] on the effect of pesticides and other chemicals on decrease in small lymphocytes due to sub-~ 2» difference in lymphocyte percentage in ot ure of Cobalt” and there was no - tee eaknent in Colisa fasciatus, In sublethal organuphosphorus peste estnen there was fluctuations in. lymphocyte percentage in Channa rnc The, lymphocytes Inthe present study finds support from a few wor . There are some reports differontial counts in fishes. There was In this sublethal treatment of Sevin to Clarias there was continuous increese in immunocyte population (lymphocytes) and decrease In phagocytic population (mono- cytes and granulocytes) of leucocytes. So the phagocytic defence mechanism was gradually decreased and the pesticide stimulated the haemopoietic organs to produce more cells of immune defence system i, e- lymphocytes. Itis also aknown fact that prolonged stimulation of immunological defence system lowers the capacity of the fish to struggle with infection which may lead to the death of the fish in long run which may happen in the present case it the sublethal toxicity is prolonged turther. In any ease the pesticide used in this study was harmful to this fish and should be used cautiously. There are some reports on the variations of TLC due to exposure in sub-lethal concentrations of pesticides and chemicals in fishes. In folidal treat ; er 5. iment TLC showed fluctuations®! in Heleopestes “fosilis, increased in weedone treatment in Pundius Ceere Aue {e Rogar Clarias batrachus™ and Anahas testdinews ®, However, TLC recorded g recorde decreased value in Heteropneustes fossilis di 08 fe ue to treatment. sublethal dichlorvos* and lindane’ Y only after 30 and 60 days of @ long period of exposure. Thus leuc i Se. Ceviosis and lymphocytosis, with respect fo ACKNOWLEDGEMENTS One of us (BK) is thay nkful to UGC for ti he grant of JRF.Effects of Sublethal Toxicity of Sevin an Blood Cav REFERENCES 4, Mahajan, C. L. and Juneja, \ ©. 4. (1979). Indian 9, Envivon, lds, 21 Pandey, P. K. Singh, N, K., Choudhury, B, P. Jish. Soe, India, 13 : 120, Dhillon, S. S. and Gupta, A. K. (1983), Water, 5162, and Thakur, GK, (1981). J. Jaland i air and Soil Pollution, 20 : 63. Srivastava, As IC. and Mishra, J, (1983). J. Comp. Path., 93: 97. Mishra, J. and Srivastava, A. K, (1983). 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