Detection of animal brucellosis by cultural method

Brucellosis, an infectious disease of many vertebrate species, is an important re-emerging zoonosis with a worldwide distribution. The present research was aimed for detection of brucellosis in animals by isolation and identification of Brucella species isolates by conventional method and PCR. Clinical specimens (61) which included vaginal swabs (48), foetal membranes (08), semen samples (03) and abomasal contents of aborted foetuses (02) were subjected for isolation on Brucella Agar Medium (BAM), wherein 15 Brucella spp. isolates were recovered with isolation rate of 24.59%. All 15 conventionally identified Brucella abortus species isolates were further confirmed by 2 different PCR assays using genus specific BCSP31 and species specific IS711 (AB and BM) primers. The BCSP31 (223 bp) PCR assay was simple, effective and sensitive in detection of Brucella isolates at genus level, whereas, IS711/AB PCR was useful to detect isolates at species level. In IS711/AB PCR, abortus specific amplification product of 498 bp was produced in all 15 isolates confirming their identity as B. abortus, whereas none of isolate showed B. melitensis specific amplification in IS711/BM PCR.