author(s)
MS Kala, LN Sankhala, Lakshmi Kant, Lalit Kumar, SR Puniya and NM Shah

abstract
Brucellosis is a widespread and economically important infectious disease of animals and humans caused by members of the genus Brucella. The present study was undertaken for isolation and identification of Brucella organisms in vaginal discharge, placental cotyledons and aborted faetal contents and detection of Brucella organisms by PCR based method of bovines of North Gujarat. Out of 107 clinical samples of bovines, Brucella could be recovered, using Brucella agar medium, from five samples. Out of these, two were from cattle while three were from buffaloes. All the five isolates were identified as Brucella organisms by cultural, morphological and biochemical characteristics and further confirmed by PCR using three different genus specific primer pairs viz. B4/F5, JPF/JPR and F4/R2.Of the 53 clinical samples tested by direct PCR, using three Brucella genus specific primer pairs, 4 were found positive by B4/B5 primer pair, 3 by JPF/JPR primer pair and 3 by F4/R2 primer pair. The B4/B5 primer pair found more suitable than other two as they resulted in highest positive numbers as well as gave all the samples positive that were positive by other two primer pairs.