Saidu A S, Singh Mahavir, Kumar Aman, Mahajan N K, Mittal Dinesh, Chhabra Rajesh, Joshi Vinay G, Musallam Imadidden I, Sadiq Usman
Department of Veterinary Public Health and Epidemiology, College of Veterinary Sciences, LUVAS, Hisar, 125004, Haryana, India.
Department of Veterinary Public Health and Preventive Medicine, Faculty of Veterinary Medicine, University of Maiduguri, P.M.B., 1069, Maiduguri, 600230, Borno State, Nigeria.
Heliyon. 2022 Feb 16;8(2):e08937. doi: 10.1016/j.heliyon.2022.e08937. eCollection 2022 Feb.
vaccines play a central role in bovine brucellosis control with tremendous success worldwide for decades. The study was aimed to evaluate the efficacy of reduced dose (5.0 × 10 9 cfu) of S19 vaccine in adult cattle and its shedding in the milk of vaccinated cattle using molecular techniques. The OIE recommended tests (RBPT, SAT, and iELISA) for brucellosis screening in cattle were used. Seronegative cattle (n = 90) of different age groups (young, old heifers & milking cows, n = 30 each) were selected for the vaccine trials. Antibody titers were recorded at 7th, 21st, 30th, 60th, 90th and 120th days post-vaccination (DPV) to monitor the immune responses following vaccination and at 150th, 180th, 210th and 240th DPB following booster-dose to an intraocular group. The humoral immune responses observed by RBPT and ELISA, proved that antibody titers persisted in s/c group compared to the i/o group in all categories. The IFN-γ stimulation (CMI) due to reduced dose vaccination was noticed early as 30th in all groups and declined after 90th DPV, with higher IFN-γ stimulation among the s/c group. The Bcsp31 and IS711 targeted PCR detected the presence of DNA in milk samples (n = 120) from the vaccinated cows (n = 30) and confirmed by qPCR (TaqMan assay) at 30th, 60th, 90th and 120th DPV. A Significant number, 70% (7/10) was detected in s/c by qPCR. BCSP31 sequence was deposited at NCBI GenBank (accession no. MK881173-6). PCR and qPCR techniques could provide a reliable diagnosis of brucellosis from milk. The intraocular route remains the safer route for vaccinating adult cattle than subcutaneous.
几十年来，疫苗在全球范围内对牛布鲁氏菌病的防控中发挥了核心作用，并取得了巨大成功。本研究旨在评估减毒剂量（5.0×10⁹ cfu）的S19疫苗在成年牛中的效力，以及使用分子技术检测接种疫苗的牛的乳汁中该疫苗的排出情况。采用了世界动物卫生组织（OIE）推荐的用于牛布鲁氏菌病筛查的检测方法（虎红平板凝集试验、试管凝集试验和间接酶联免疫吸附试验）。选择不同年龄组（幼年、老年小母牛和泌乳奶牛，每组n = 30头）的血清阴性牛（n = 90头）进行疫苗试验。在接种疫苗后第7、21、30、60、90和120天记录抗体滴度，以监测接种疫苗后的免疫反应，并在对眼内接种组进行加强剂量接种后的第150、180、210和240天记录抗体滴度。通过虎红平板凝集试验和酶联免疫吸附试验观察到的体液免疫反应证明，在所有类别中，与眼内接种组相比，皮下接种组的抗体滴度持续存在。由于减毒剂量接种导致的γ干扰素刺激（细胞介导免疫）在所有组中早在第30天就被注意到，并在接种疫苗后第90天下降，皮下接种组中的γ干扰素刺激更高。以Bcsp31和IS711为靶点的聚合酶链反应（PCR）检测了接种疫苗的奶牛（n = 30头）乳汁样本（n = 120份）中DNA的存在，并在接种疫苗后第30、60、90和120天通过实时荧光定量PCR（TaqMan检测法）进行了确认。通过实时荧光定量PCR在皮下接种组中检测到相当数量，即70%（7/10）。BCSP31序列已保存在美国国立生物技术信息中心（NCBI）的基因库中（登录号：MK881173 - 6）。PCR和实时荧光定量PCR技术可为从乳汁中诊断布鲁氏菌病提供可靠的诊断。对于成年牛接种疫苗而言，眼内接种途径比皮下接种途径更安全。
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