Background and Aim: Flinders Technology Associates (FTA) cards simplify sample storage, transport, and extraction by reducing cost and time for diagnosis. This study evaluated the FTA suitability for safe transport and storage of Gram-positive and Gram-negative bacterial cells of animal origin on its liquid culture form and from organ impression smears (tissues) under the same routine condition of microbiological laboratory along with detecting their nucleic acid over different storage conditions. Materials and Methods: Increase in bacterial count from 104 to 107 (colony-forming units/mL) of 78 isolates representing seven bacterial species was applied onto cards. FTA cards were grouped and inoculated by these bacteria and then stored at different conditions of 24-27°C, 4°C, and –20°C for 24 h, for 2 weeks, for 1 and 3 month storage, respectively. Bacteriological examination was done, after which bacterial DNA was identified using specific primers for each bacterial type and detected by polymerase chain reaction (PCR). Results: The total percentage of recovered bacteria from FTA cards was 66.7% at 24-27–C for 24 h, the detection limit was 100% in Gram-positive species, while it was 57.4% in Gram-negative ones. Regarding viable cell detection from organ impression smears, it was successful under the previous conditions. No live bacterial cells were observed by bacteriological isolation rather than only at 24-27°C for 24 h storage. All bacterial DNA were sufficiently confirmed by the PCR technique at different conditions. Conclusion: Overall, the FTA card method was observed to be a valid tool for nucleic acid purification for bacteria of animal origin in the form of culture or organ smears regardless of its Gram type and is used for a short time only 24 h for storage and transport of live bacteria specifically Gram-positive type. Moreover, the bacterial nucleic acid was intact after storage in –20°C for 3 months and was PCR amplifiable.
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.
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