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印度查谟和克什米尔地区绵羊腐蹄病中结节拟杆菌的流行率、血清学多样性及毒力测定,并鉴定其主要血清型作为潜在疫苗候选株

Determination of prevalence, serological diversity, and virulence of Dichelobacter nodosus in ovine footrot with identification of its predominant serotype as a potential vaccine candidate in J&K, India.

作者信息

Wani Shakil A, Farooq S, Kashoo Z A, Hussain I, Bhat M A, Rather M A, Aalamgeer S

机构信息

Bacteriology Laboratory, Division of Veterinary Microbiology and Immunology, SKUAST-K, Shuhama (Alusteng), Srinagar, Jammu and Kashmir, 190006, India.

Division of Veterinary Microbiology and Immunology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu (SKUAST-J), R.S. Pura, Jammu, Jammu and Kashmir, 181102, India.

出版信息

Trop Anim Health Prod. 2019 Jun;51(5):1089-1095. doi: 10.1007/s11250-018-01788-9. Epub 2019 Jan 9.

Abstract

The aim of this study was to determine the prevalence, serological diversity, and virulence of Dichelobacter nodosus in footrot lesions of sheep and identification of its predominant serotype as a potential vaccine candidate. The overall prevalence of footrot in sheep was 16.19%, and ranged from 13.69 to 19.71%, respectively. A total of 759 flocks with 22,698 sheep were investigated for footrot and 2374 clinical samples were collected from naturally infected sheep exhibiting footrot lesions. Of the 2374 samples collected, 1446 (60.90%) were positive for D. nodosus by polymerase chain reaction (PCR). These positive samples when subjected to serogroup-specific multiplex PCR, 1337 (92.46%) samples carried serogroup B, 247 (17.08%) possessed serogroup E, 86 (5.94%) serogroup I, and one (0.069%) serogroup G of D. nodosus. While mixed infection of serogroups B and E was detected in 127 (8.78%), B and I in 46 (3.18%) and B, E, and I in 26 (1.79%) samples, respectively. The serogroup B of D. nodosus was the predominant (92.47%) serogroup affecting sheep population with footrot followed by serogroup E (19.91%) and serogroup I (4.57%), respectively. Virulent status of D. nodosus strains were confirmed by presence of virulence-specific integrase A (intA) gene and the production of thermostable proteases. The intA gene was detected in 709 (72.79%) samples while gelatin gel test carried out on 246 representative isolates all positive for intA gene produced thermostable proteases, confirming their virulence nature. The PCR-restriction fragment length polymorphism (PCR-RFLP) of whole fimA gene of serogroup B revealed the predominance of serotype B5 (82.97%) of serogroup B. This information suggests that serotype B5 is the predominant serotype of D. nodosus associated with severe footrot lesions in sheep in Jammu & Kashmir (J&K), India. Hence, this serotype can be a potential vaccine candidate for the effective control and treatment of ovine footrot.

摘要

本研究的目的是确定绵羊腐蹄病病变中结节拟杆菌的流行率、血清学多样性和毒力,并鉴定其主要血清型作为潜在的疫苗候选物。绵羊腐蹄病的总体流行率为16.19%,分别在13.69%至19.71%之间。共调查了759个羊群中的22698只绵羊的腐蹄病情况,并从表现出腐蹄病病变的自然感染绵羊中收集了2374份临床样本。在收集的2374份样本中,通过聚合酶链反应(PCR)检测到1446份(60.90%)结节拟杆菌呈阳性。对这些阳性样本进行血清群特异性多重PCR检测时,1337份(92.46%)样本携带结节拟杆菌血清群B,247份(17.08%)携带血清群E,86份(5.94%)携带血清群I,1份(0.069%)携带血清群G。同时,分别在127份(8.78%)样本中检测到血清群B和E的混合感染,46份(3.18%)样本中检测到血清群B和I的混合感染,26份(1.79%)样本中检测到血清群B、E和I的混合感染。结节拟杆菌血清群B是影响患腐蹄病绵羊群体的主要血清群(92.47%),其次是血清群E(19.91%)和血清群I(4.57%)。通过毒力特异性整合酶A(intA)基因的存在和耐热蛋白酶的产生来确认结节拟杆菌菌株的毒力状态。在709份(72.79%)样本中检测到intA基因,而对246株代表菌株进行明胶凝胶试验,所有intA基因呈阳性的菌株均产生耐热蛋白酶,证实了它们的毒力性质。血清群B全fimA基因的PCR-限制性片段长度多态性(PCR-RFLP)显示血清群B中血清型B5占主导地位(82.97%)。该信息表明,血清型B5是印度查谟和克什米尔(J&K)地区与绵羊严重腐蹄病病变相关的结节拟杆菌的主要血清型。因此,该血清型可作为有效控制和治疗绵羊腐蹄病的潜在疫苗候选物。

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