The plate counting method widely used at present to discern viable from non-viable Brucella in the host or cell is time-consuming and laborious. Therefore, it is necessary to establish a rapid, simple method for detecting and counting viable Brucella organisms.

The plate counting method widely used at present to discern viable from non-viable Brucella in the host or cell is time-consuming and laborious. Therefore, it is necessary to establish a rapid, simple method for detecting and counting viable Brucella organisms. Using propidium monoazide (PMA) to inhibit amplification of DNA from dead Brucella, a novel, rapid PMA-quantitative PCR (PMA-qPCR) detection method for counting viable Brucella was established. The standard recombinant plasmid with the target BCSP31 gene fragment inserted was constructed for drawing a standard curve. The reaction conditions were optimised, and the sensitivity, specificity, and repeatability were analysed. The optimal exposure time and working concentration of PMA were 10 min and 15 μg/mL, respectively. The correlation coefficient (R²) of the standard curve was 0.999. The sensitivity of the method was 10³ CFU/mL, moreover, its specificity and repeatability also met the requirements. The concentration of B. suis measured by the PMA-qPCR did not differ significantly from that measured by the plate counting method, and the concentrations of viable bacteria in infected cells determined by the two methods were of the same order of magnitude. In this study, a rapid and simple PMA-qPCR counting method for viable Brucella was established, which will facilitate related research.

Objective: The study aimed to explore the seroprevalence of some bacterial (Brucella spp., Chlamydia abortus) and viral [Rift Valley fever virus (RVFV), foot and mouth disease virus (FMDV)] zoonoses in domestic ruminants in Medina. Materials and Methods: A total of 1,000 blood samples from 665 sheep, 228 goats, and 107 camels were collected from the central slaughterhouse, private farms, and veterinary clinics affiliated to the Ministry of Agriculture. The samples were screened using the Enzyme-Linked Immunosorbent Assay (ELISA). The results were statistically analyzed using Statistical Package for the Social Sciences version 20. Results: Brucella was found in 7.7%, 8.8%, and 6.5% of sheep, goats, and camels sera, respec¬tively. In humans, Brucella abortus and Brucella melitensis had higher frequencies in males (61.3%, 59.8%) than in females (38.7%, 40.2%). Chlamydia abortus was reported only in sheep at a rate of 0.75%. RVFV was prevalent in sheep (10.7%) and goats (17.9%). FMDV was reported in 27.8% of sheep and 7.9% of goats. There was a highly statistical significance between Brucella, RVFV, and FMDV seropositivity and locally bred animals (p < 0.01). Multiple seropositivities have been reported among sheep and goats. Brucella was commonly observed in mixed infection with other bacterial and viral agents under study. Conclusion: The surveyed viral and bacterial agents were prevalent in ruminants in the Medina region. Since Medina is an important destination for pilgrims from all over the world, therefore, an integrated approach involving strict control measures and routine vaccination programs should be adopted to reduce the possibility of global epidemics. [J Adv Vet Anim Res 2020; 7(1.000): 42-50]