Brucellosis is considered an economically important highly contagious and zoonotic bacterial disease of water buffaloes. Control of brucellosis in buffaloes is very important for public health. The efficacy of control program depends on the detection and eradication of infected animals coupled with vaccination and application of biosecurity. This study was carried out to control the brucellosis in buffalo farm in Assuit Governorate, Egypt during the period from April 2015 to August 2016. Out of 620 unvaccinated buffaloes, 87 (14.03%) aborted. Moreover, 90/620(14.51%), 82/620(13.22%), 82/620(13.22%), and 80/620 (12.9%) buffaloes were serologically positive by BAPA, RBPT, m SAT and Riv.T, respectively. Three isolates were differentiated as Brucella melitensis, biovar 3, one strain isolated from one vaginal swap out of 10 Riv.T. positive recently aborted buffaloes (10%) and two strains were isolated out of ten milk samples of Riv.T. positive buffaloes (20%). Eighty serological positive buffaloes to Riv.T were culled from the herd, while 60 serological negative heifers were vaccinated by Brucella abortus S 19 vaccine, with a dose of 3-8×109 cfu/5ml and monitored for serological titer for 240 days. After 6 months of vaccination, the number of serologically positive calves declined marginally to 50 (83.33%), 40 (66.67%), 50 (83.33%), 0 (0%), 40 (66.67%) and 0 (0%) by BAPA, RBPT, mSAT, CFT, iELISA and cELISA, respectively. Three successive serological tests every three weeks were done by screening tests, BAPA and RBPT and confirmed by Riv.T. At the end of the control program, all examined buffaloes were serologically negative. Application of biosecurity in the farm was applied by the sanitary disposal of aborted material and application of proper disinfectants at its recommended work strength and contact time.
Brucellosis is one of the world's major zoonoses that still has veterinary, public health and economic concern in many parts of the world. In livestock, brucellosis is the major impediment for trade and export. In this work a total of 118 different samples (45 serum samples, 45 blood samples, stomach contents of 18 aborted feati and 10 fecal samples) were collected from 45 diseased and apparently healthy cows at selected veterinary clinics at El-Sharkia governorate. The serum samples were subjected to investigation by different serological methods (Rose Bengal, BAPAT, CFT and ELISA) for detection of bovine brucellosis. While, molecular differentiation between Brucella and Yersenia enterocolitica O:9 were made on blood samples, stomach contents and fecal samples of sero positive animals for brucellosis using multiplex PCR assay. The results revealed that 26 out of 45 serum samples were positive for brucellosis, as detected by ELISA while 19 samples gave positive amplification of 223bp for Brucella and seven positive amplification of 325bp for Yersinia representing 26.92% of total brucellosis seropositives of which two samples (7.69%) were positive for both Yersinia and Brucella as tested by multiplex PCR assay. Therefore, we concluded that multiplex PCR proved to be a reliable molecular method for differentiation between Brucella and Yersinia as rapid diagnostic tool directly from clinical specimens.
Background and Aim: Different polymerase chain reaction (PCR) techniques have and are still being used for the direct detection of Brucella DNA in serum samples of different animal species and humans without being validated or properly validated, resulting in discrepancies. Thus, this study aimed to evaluate the diagnostic performance of the TaqMan Real- Time-PCR (RT-PCR) targeting the bcsp31 gene versus conventional PCR for the accurate diagnosis of brucellosis at the genus level in cattle sera. Materials and Methods: One hundred and eighty-four serum samples were collected from bacteriologically positive and negative cows with ages ranging from 1 to 5 years old at some infected private farms in the Nile Delta under quarantine measures as well as brucellosis free farms. These samples were classified into four groups after serological diagnosis and investigated by TaqMan RT-PCR and conventional PCR targeting the IS711 gene for Brucella DNA detection. The diagnostic performance characteristics of both PCR techniques were estimated considering the bacteriological results as a gold standard. Results: TaqMan RT-PCR revealed superiority over conventional PCR; it was able to detect Brucella DNA in 95% (67/70) and 89% (25/28) of the cattle sera samples belonging to Group 1 (serologically and bacteriologically positive) and Group 2 (serologically negative but bacteriologically positive), respectively. On evaluating the diagnostic performance, TaqMan RT-PCR showed superior diagnostic sensitivity (93.9%), diagnostic specificity (88.4%), performance index (182.3), almost perfect kappa agreement (0.825±0.042), strong positive correlation (r=0.826), high accuracy based on the receiver operating characteristic (ROC) curve, and area under the ROC curve (0.911) at p<0.05 and CI of 95%. Conclusion: A cattle serum sample is not the metric of choice for targeting Brucella genomic DNA by conventional PCR. The time-saving and rapid TaqMan RT-PCR method revealed a better diagnostic performance in the detection of Brucella DNA in cattle sera. Such performance offered by TaqMan RT-PCR may be considered a step toward the possibility of using such technology in the direct differentiation between Brucella-infected and -vaccinated cattle immunized by smooth vaccines from cattle sera using primers specific for such vaccines.
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