ELISA BASED SURVEILLANCE OF BOVINE FASCIOLIASIS IN DISTRICT SARGODHA

Abstract

An excretory/ secretory antigen based ELISA was developed for diagnosis of bovine fascioliasis and evaluated in field survey in district Sargodha in comparison with coprological examination and commercial kit (DRG Diagnostica). The study was divided into three experiments. First experiment was slaughter house based epidemiology of bovine fasciolasis. Livers, bile ducts and fecal samples of 146 cattle and 184 buffaloes were examined for presence of Fasciola from slaughter house of district Sargodha. Significantly higher prevalence was found in buffaloes as compared to cattle in both coprological (χ2=25.312; P-value=0.0080) and fecal examination (χ2=17.453; P value=0.0001). By coprological examination, Fasciola eggs were detected in 29.89% (55/184) buffaloes and in 20.55% (30/146) cattle whereas 48.37% (89/184) buffaloes and 37.67% (55/146) cattle were found infected during liver examination. Sex was found to be non-significantly associated with risk of infection while negative association was found between age and occurrence of disease. Prevalence of disease was lower in higher age groups. In second experiment, development and standardization of ES antigen based ELISA was conducted. Adult Fasciola and blood samples were collected from 146 cattle and 184 buffaloes. Excretory/ secretory antigens were prepared by incubating adult Fasciola in PBS for 6-8 hrs and then filtering through 0.22 μm syringe filter. Then checker board titration was performed and optimum concentration of antigen and serum was determined and used in screening of all serum samples. Seroprevalence was found to be 50% and 38.35% in buffaloes and cattle, respectively. In house ELISA was found to be 100 % sensitive while specificity was 96.84% in buffaloes and 98.90% in cattle. In third experiment, serological and coprological survey of bovine fascioliasis of district Sargodha was carried out. A total of 600 blood samples and 5580 fecal samples were collected from all six tehsils of district Sargodha. Sedimentation- floatation technique was adopted to identify Fasciola egg in feces. Sera were screened for presence of antifasciola antibodies by indigenous ELISA kit and DRG kit. Both kits were equally sensitive while indigenous ELISA was more specific. The highest prevalence was found during month of December in both serological and coprological examination. Higher prevalence was recorded in tehsil Bhalwal, Sahiwal and Shahpur as compared to Sargodha, Kot-Momin and Silanwali. Risk of fascioliasis was found to be negatively associated (OR=1.181; χ2=105.6757; P-value <0.0001) with age categories, highest prevalence of fascioliasis in >2-4 years age group and then decreasing with advancing age. Sex was found to be non-significantly associated with disease. As for managemental practices are concerned, higher prevalence was found in grazing group (χ2=61.3443; P-value <0.0001), pond watered and river watered group (χ2=89.7096; P-value <0.0001) as compared to stall feeding and tap watered group.