Isolation and molecular detection of brucella melitensis from disease outbreak in sheep and b. abortus from cattle farm by is711 and omp2a gene based pcr

Brucellosis is one of the world’s major zoonoses that still is of veterinarian, public health and economic concern in many parts of the world. Brucellosis due to Brucella melitensis is of much public health and economic importance in many developing countries including India and is considered to be the major cause of abortion in small ruminants. The present study was carried out with the objective of cultural isolation of B. melitensis from clinical samples (knee joint fluid and aborted foetal material from 56 animals) collected from a disease outbreak in sheep in Saharanpur District (U.P., India) and its confirmatory detection using molecular tool of polymerase chain reaction (PCR). Out of a total number of 56 clinical samples, 42 (75%) bacterial isolates of Brucella spp. were recovered. On the basis of colony morphology, staining characters, phenotypic and biochemical characterizations, the organisms from clinically infected sheep identified as B. melitensis. Further confirmation of B. melitensis done by PCR amplification of IS711 and omp2a target genes gave specific amplicons of 731 bp and 1104 bp fragment sizes, respectively for all the 42 cultural isolates obtained. In addition, using both cultural methods and PCR, 02 (33.33%) isolates of B. abortus were also isolated and identified from liver samples of aborted bovine foetus (n=6) collected from an organized Cattle farm of Bareilly (U.P.). Isolation and confirmatory diagnosis of B. melitensis and B. abortus indicates appropriate prevention and control strategies for this economically important pathogen having zoonotic significance.