Aim::Brucellosis is a major bacterial zoonosis of global importance affecting a range of animal species and man worldwide. It has economic, public health, and bio-risk importance. Control and prevention of animal brucellosis mainly depend on accurate diagnostic tools and implementation of effective and safe animal vaccination program. There are three types of animal Brucella live vaccines - Brucella melitensis Rev-1 vaccine, Brucella abortus S19, and B. abortus RB51. Evaluation of these vaccines depends mainly on enumeration of Brucella viable count. At present, used colony count method is time consuming, costly and requires especial skills. Hence, the aim of this study is to use and standardize real-time polymerase chain reaction (RT-PCR) as an alternative, quantitative, sensitive, and rapid method to detect the colony count of Brucella in live Brucella vaccine.Materials and Methods::Four batches of different live Brucella vaccines were evaluated using of conventional bacterial count and RT-quantitative PCR (RT-qPCR) using BSCP31 gene specific primers and probe. Standard curve was generated from DNA template extracted from 10-fold serial dilution of living B. abortus RB51 vaccine to evaluate the sensitivity of RT-qPCR.Results::Results revealed that three batches of living Brucella vaccines were acceptable for Brucella colony count when traditional bacterial enumeration method was used. Results of RT-qPCR were identical to that of conventional bacterial count.Conclusions::Results concluded that RT-qPCR was relatively sensitive compared to traditional bacterial colony count of these vaccines.
Pigeon paramyxovirus type 1 (PPMV-1) and salmonellosis are two of the major health problems that affect pigeons worldwide. Immuno-protection of pigeons against PPMV-1 infection and salmonellosis is a very important preventive measure. In the present work, a combined inactivated montanide ISA-206 oil adjuvanted vaccine of local isolates of PPMV-1 and S. Typhimurium was prepared. Quality control assessment of such preparatıon revealed that it is free from foreign contaminants, safe and immunogenic. The sero-evaluation using microplate agglutination test revealed that the humoral immune response developed against S. Typhimurium in the vaccinated pigeons reached 64 three weeks post 1 st dose and reached its maximum value 256 two weeks post boostering. Results of HI test showed that the vaccine induced detectable humoral immune response to PPMV-1 expressed by marked increased HI antibody titer till the end of the experiment (8.0 log 2). The vaccination-challenge assay with the virulent strain of PPMV-1 with 10 6 EID 50 /mL showed 100% protection in vaccinated group (Ia). While, the virulent S. Typhimurium organism with 5 x10 7 CFU/bird showed 90% protection in vaccinated birds. The unvaccinated control group showed 10% and 20% protection against both virulent PPMV-1 and S. Typhimurium, respectively. In conclusion, this vaccine could be recommended as safe, potent and could be useful when used for protection against PPMV-1 and S. Typhimurium under field condition.
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