Mhase, P. P.Walunj, T. D.2017-06-222017-06-222015http://krishikosh.egranth.ac.in/handle/1/5810022696Brucellosis is an important infectious disease of livestock affecting a wide range of animal species including human and is characterized by abortions, fetal death, reproductive tract complications and arthritis in animals with great losses. Early precise diagnosis with more sensitive and specific test is a very important for its control and eradication. For the diagnosis of brucellosis different serological tests are used widely, but often they come with certain disadvantages like longevity and lack specificity and gold standard isolation cultural test requires special biosecurity facilities and poses a danger of infection. Hence, highly sensitive genus specific molecular techniques are preferred. Hence present studies dealt with the screening of farms having clinical brucellosis with serological tests; RBPT and iELISA followed by the PCR assay. Total 401 recently aborted and ‘In-contact’ animals were screened with RBPT belonging to the organized and unorganized farms of Pune and Ahmednagar region out of whihc 200 samples each were processed by iELISA employing two kits, one indigenous (ELISA-1) and another was commercial (ELISA-2). The serum and whole blood samples (24 each) were processed with genus specific BCSP 31 PCR for detection of brucellosis and comparative evaluation of serological tests. With the RBPT, ELISA-1 and ELISA-2 the percent seropositivity in cattle of farms in Pune region was 27.18%, 42.00% and 48.00% respectively, while from the Ahmednagar region it was 36.76%, 44.00% and 50.00% respectively, thus seropositivity observed was 30.99% , 43.00% and 49.00%, respectively. The percent seropositivity detected in Buffaloes of Pune region was 35.71%, 45.00% and 47.50% respectively, while from the Ahmednagar region it was 41.48%, 48.33 and 50.00% respectively, hence total was 40.08%, 47.00% and 49.00%, respectively. With RBPT total percent seropositivity was 36.40%, with ELISA-1 it was 45.00% and with ELISA-2 it was 49.00% in buffaloes. The percent seropositivity for brucellosis in the cattle of organized farm of Pune indicated 25.86%, 41.37% and 48.27% while on the unorganized farm it was 28.88%, 42.85% and 47.61%, respectively. Similarly in the cattle from Ahmednagar district seropositivity detected on organized farms was 35.55%, 50.00% and 59.37% while in unorganized farms it was 39.13%, 33.33% and 33.33%, respectively. Overall seropositivity in cattle from organized farms was 30.09%, 45.90% and 54.09% and in unorganized farms was 32.35%, 38.46% and 41.02% respectively, thus overall in cattle it was 30.99%, 43.00% and 49.00%, respectively. The percent seropositivity in buffaloes of Pune organized farms was 34.61%, 42.30% and 46.15% while in unorganized it was 37.50%, 50.00% and 50.00%, respectively. Percent seropositivity in organized farm buffaloes was 39.86%, 46.15% and 49.23% respectively, while unorganized farms it was 41.66%, 48.57% and 48.50 % respectively. Hence, total was 40.08%, 47.00% and 49.00%, respectively. The aborted cattle and buffaloes with of Pune serologically were 42.85% and 66.66%, 72.72% and 66.66%, 81.81% and 55.55%, while Ahmednagar region were 47.61% and 34.54, 53.33% and 66.66 %, 73.33% and 77.77%, positive respectively. The In-contact cattle and buffaloes of Pune were 23.17% and 20.00%, 33.33% and 38.07%, 38.46% and 45.16% while of Ahmednagar were 31.91% and 44.36%, 40.00% and 39.53 %, 40.00% and 38.09%, positive, respectively. It was concluded that brucellosis was more in animals belonging to Ahmednagar than Pune, was higher in buffaloes than in cattle, was more in the aborted than In-contact animals and higher in animals of organized farms than unorganized farms. The seropositivity was marginally higher in ELISA-2 kit than Indigenous ELISA-1 followed by RBPT. A 223 bp amplification product was observed in 30/48 samples with genus specific BCSP31PCR out of which 62.50% each of cattle and buffaloes’ samples yielded positive. The proportion of positive cases in aborted animals was higher (82.14%) than ‘In-contact’ (44.44%) cases. In serum (66.66%) positivity was higher than in blood (58.33%) samples. In aborted (91.66) cases positivity was higher than In-Contact cases (33.33%). The positivity in Ahmednagar (70.83%) was higher than in Pune(54.16%). The results of PCR compared with RBPT revealed cent percent sensitivity (100%) but the specificity was 83.33% and agreement was 90.91 %. ELISA-1 revealed sensitivity of 90.00% but cent percent (100%) specificity and 90.00% agreement while ELISA-2 presented best results with cent percent specificity (100%) and 96.66% sensitivity with 96.67% agreement with PCR in detecting Brucellosis.ennullThesis