Isolation of Brucella melitensis from seronegative camel: potential implications in brucellosis control

•The first report of Brucella melitensis isolation from seronegative camel.•Infected camel may increase the risk of brucellosis within human and livestock.•There are moderate agreements between RBT, Wright and 2ME serological results.•PCR-based methods represent a powerful approach for camel brucellosis screening. Brucellosis is a significant zoonotic infection in Iran impacting both humans and animal health status. A number of reasons such as few clinical signs complicate the diagnosis of this infection in Camelidae. Despite the ubiquitous use of serological tests for the first screening of brucellosis in camel, this approach showed several restrictions because of the intracellular properties of this organism as well as decline antibody titers in chronic stage. This study aimed at identifying the presence of Brucella spp. in blood and lymph node samples collected from slaughtered male camels of Sistan-Baluchistan province by serology, culture and polymerase chain reaction (PCR) assay. For this purpose, 2854 blood camel samples were sampled and analyzed for Brucella detection by serological screening tests. The molecular detection of IS711 gene and Bruce-ladder assay as well as culture were performed using the lymph nodes of all seropositive camel (n = 10) and 30 seronegative samples. Results showed that 0.35% (10/2854), 0.24% (7/2854) and 0.21% (6/2854) of blood samples were positive by RBPT, SAT and 2ME, respectively. However, 0% (0/10) and 70% (7/10) of lymph node samples collected from seropositive camels were positive for Brucella infection by culture and PCR, respectively. Furthermore, 6.6% (2/30) of seronegative lymph node specimens showed the presences of Brucella by PCR and culture assay. The results of the present study indicated the low seroprevalence of Brucella infection in male camels of the Sistan-Baluchistan province and highlighted the complementary role of PCR techniques for a better screening of Brucella infection among seronegative camels. Moreover, the potential shedding of Brucella within undiagnosed camel milk and secretions is a serious problem which may result in further spread and maintenance of Brucella infection among both human and livestock. Thus, for brucellosis detection and control, our results suggested that a first PCR screening supported by a bacteriological isolation on positive samples should be performed along with the serological test in endemic countries to identify the source and prevent the uncontrolled spread of