STUDIES ON CLINICO-PATHOLOGICAL AND IMMUNOMODULATORY EFFECTS OF COW URINE AND ITS SYNERGISTIC ACTION WITH AQUEOUS POLY HERBAL EXTRACT IN COMMERCIAL LAYER CHICKS
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Date
2013
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AAU, Anand
Abstract
The present investigation was undertaken to study on clinico-pathological and immunomodulatory effects of cow urine distillate and its synergistic action with aqueous poly herbal extract containing Ocimum sanctum (Tulsi), Tinospora cordifolia (Guduchi) and Withania somnifera (Ashwagandha) in commercial layer chicks. A total of 270, one day old BV-300 layer chicks were randomly divided into nine groups comprising 30 chicks in each group. Group I served as negative control without administration of any treatment except plain water at ad libitum. Groups II and III were served as positive treatment control groups and administered with IBD intermediate vaccine and cyclophosphamide as immunosuppressive agents respectively. The remaining treatment groups IV to IX were given either cow urine distillate alone or its combination with aqueous poly herbal extract along with intermediate IBD vaccine and cyclophosphamide. All the birds of different experimental groups were observed for different parameters. Six birds from each experimental group were sacrificed on day 0, 15, 30, 45 and 60 for collection of material for haematological, biochemical, histopathological and lymphoblastogenesis study. Clinical signs like tendency to remain standing at one place with apathy, unthriftiness with ruffled feathers, dullness and drooping of the wings were observed in some of the birds belonged to group II (IBD vaccinated) and III (cyclophosphamide) after two to three days of administration of immunosuppressive agents. Except mild depression and dullness in occasional birds from treatment groups VI, VII, VIII & IX, no other clinical signs were observed in other treatment groups throughout the study period. There was significant (P<0.05) increase in body weight in groups IV and V as compared to group I as well as in groups VI and VII as compared to group II and in group VIII and group IX as compare to group III on 30th and 45th days of experiment. There was significant increase (P<0.05) in bursa body weight ratio and thymus body weight ratio in groups IV (cow urine distillate ) and V (cow urine distillate + aqueous poly herbal extract) on 15th and 30th days as compared to control group I. There was significant decrease (P<0.05) in bursa body weight ratio and thymus body weight ratio in groups II and III as compared to control group I on 30th day of experiment. There was significant increase (P<0.05) in both these ratios in groups VI and VII as compared to group II and in groups VIII and IX as compared to group III on 30th day of experiment. Different haematological parameters revealed significant decrease (P<0.05) in haemoglobin (Hb), packed cell volume and TEC count in group III on 15th and 30th days of experiment as compared to control group I. Among different treatment groups, only groups VIII and IX showed significant decrease (P<0.05) in Hb, PCV and TEC values in comparison to group I and significant increase as compared to group III on 15th day of experiment. There was significant decrease (P<0.05) in the TLC in groups III, VIII and IX as compared to group I whereas significant increase (P<0.05) in groups VIII and IX as compared to group III on 15th day of experiment. There was also significant decrease in heterophil, lymphocyte and monocyte counts in group III on 15th and 30th days as well as in groups VIII and IX on 15th day as compared to group I where as the values of heterophil and lymphocyte counts were significantly higher in groups VIII and IX as compared to group III. All these findings indicated that cow urine and its combination with poly herbal extract have ability to counteract the effect of cyclophosphamide in layer chicks. Among different biochemical parameters studied, ALT and AST values revealed significant increase in group III as compared to group I while significant decrease was observed in groups VIII and IX as compared to group III on 15th and 30th days of experiment. There was significant increase in BUN and creatinine in group III on 15th and 30th days as well as in groups VII and IX on 15 day as compared to group I where as significant decrease in groups VIII and EX as compared to group III on 15 and 30th days of experiment. There was significant decrease (P<0.05) in NDHI antibody titre in group II (IBD vaccinated) on 30th, 45th and 60th days of experiment as well as in group III (cyclophosphamide) on 30th day of experiment as compared to control group I. There was significant increase (P<0.05) in antibody titre in groups IV, V, VI, VII, VIII and IX as compared to their respective control groups on 30th , 45th and 60th days of experiment. Mean values of skin thickness revealed significant increase in groups IV, V, VI, VII, VIII & IX as compared to group I, in groups VI and VII as compared to group II and in groups VIII and IX as compared to group III at 24 hrs after DNCB application. Lymphoblastogenesis assay revealed significant decrease (P<0.05) in T-cell blastogenesis in group III and B-cell blastogenesis in groups II and III as compared to group I on 30th and 45th days of experiment. There was significant (P<0.05) increase (P<0.05) in T-cell and B-cell blastogenesis in groups IV and V as compared to group I, in groups VI and VII as compared to group II and in groups VIII and IX as compared to group III on 30th , 45th and 60th days of experiment. These results suggested the potent humoral and cell mediated immune response of cow urine distillate and its combination with poly herbal extract. There was marked gross enlargement of bursa of Fabricius and thymic lobes in all the treatment groups IV, V, VI, VII, VIII and DC on 30th and 40th days of sacrifice when compared to their respective control groups. There were no appreciable gross lesions in other internal organs. Variable lesions like erythema, indurations, haemorrhage and necrosis were seen in groups IV to IX at 24 his post challenge of DNCB application. Microscopic lesions in bursa of Fabricius revealed enlarged bursal follicles with lymphoid hyperplasia in groups IV and V as compared to group I on 30th and 45th days of sacrifice. There was mild to moderate atrophy of lymphoid follicles in groups II and III while in comparison to both these groups, the bursal follicles of birds belonging to groups VI, VII, VIII and IX showed mild to moderate regeneration in the form of lymphoid cell proliferation on 30th and 45th days of sacrifice. Lesions in the thymus revealed slightly enlarged lymphoid follicles with lymphoid hypeiplasia in groups IV and V on 30 and 45th days of sacrifice. Thymic lesions in group III treated with cyclophosphamide mainly consisted mild to moderate atrophy with depletion of lymphocytes while moderate enlargement of thymic follicles with regenerative proliferation of lymphocytes was observed in groups VIII and IX as compared to group III on 30th and 45th days of experiment. Sections of spleen revealed mild depletion of lymphocytes in group III on 30th and 45th days of sacrifice while there was regenerative RE cell hyperplasia in groups VIII and IX on 30 day of experiment. Section of kidney showed lesions only in group III. The kidney lesions were characterized by mild congestion, widespread degeneration and necrosis of tubular epithelium. The severity of renal lesions was markedly reduced in groups VIII and IX as compared to group III on 30th day of sacrifice. The histopathological lesions in the skin after 24 lirs of DNCB application revealed lesions like varying degrees of mononuclear cell infiltrations accompanied with necrosis, edema and congestion in superficial and deep layers of dermis in various groups treated with cow urine alone or its combination with poly herbal extract. It could be concluded from the present findings that cow urine distillate alone or its mixture with poly herbal aqueous extract was found to modulate the immunity in a positive maimer and can be used as an inamunomodulatory agents in commercial layer chicks.
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VETERINARY PATHOLOGY, A STUDY