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埃塞俄比亚中部选定地区绵羊肺炎病例相关溶血曼海姆菌分离株的分子特征。

Molecular characterization of Mannheimia haemolytica isolates associated with pneumonic cases of sheep in selected areas of Central Ethiopia.

机构信息

National Veterinary Institute, P.O. Box 19, Bishoftu, Ethiopia.

College of Veterinary Medicine and Agriculture, Addis Ababa University, P.O. Box 34, Bishoftu, Ethiopia.

出版信息

BMC Microbiol. 2018 Dec 5;18(1):205. doi: 10.1186/s12866-018-1338-x.

Abstract

BACKGROUND

Mannheimia haemolytica has been recognized as the principal cause of pneumonic pasteurellosis in sheep and goats. It is one of the important diseases of small ruminants in Ethiopia. While annual vaccination using a monovalent vaccine (inactivated Pasteurella multocida biotype A) is common, respiratory diseases are still reported in various parts of Ethiopia. This suggests the need for further investigation into the species and strains responsible for the disease, which is vital information for development of a multivalent vaccine. The objective of the current study was to isolate M. heamolytica associated with pneumonic cases of sheep in selected areas of Central Ethiopia, determine its role and the strains/genotypes of the bacterium circulating in the study area.

RESULTS

Bacteriological analysis of nasal swab samples collected from a total of 76 pneumonic cases of sheep showed that M. haemolytica was isolated from 26 of them while B.trehalosi from two cases. Further molecular analyses of the isolates using M. haemolytica species-specific and M.haemolytica serotype-1 antigen specific PCR assays revealed, 26 of the isolates were identified as M. haemolytica of which 21 of them were M. haemolytica serotype-1. Both M. haemolytica and B.trehalosi isolates were not detected in a PCR assay targeting capsular biosynthesis gene (capA) of P.multocida despite the non-specific products observed in M. haemolytica isolates. Phylogenetic analysis of M. haemolytica isolates included in this study in comparison with the reference strains with respect to PHSSA and Rpt2 genes revealed that the Ethiopian M. haemolytica isolates constituted three distinct genotypes consistent with site of origin.

CONCLUSION

The study indicated that M.haemolytica is commonly associated with cases of pneumonia in sheep in the study areas of central Ethiopia although the remaining other pathogens responsible for majority of the cases are yet to be determined. Molecular characterization revealed the existence of three genotypes of M. haemolytica circulating in the study areas consistent to the site of isolation. The findings suggest further extensive work to determine all pathogens associated with sheep pneumonia and the strain distribution of M. heamolytica to understand its molecular epidemiology at national level and design cost effective prevention and control methods.

摘要

背景

曼海姆菌已被确认为绵羊和山羊肺炎巴氏杆菌病的主要病原体。它是埃塞俄比亚小反刍动物的重要疾病之一。虽然每年使用单价疫苗(灭活多杀性巴氏杆菌生物型 A)进行接种很常见,但在埃塞俄比亚的不同地区仍有呼吸道疾病的报道。这表明需要进一步调查导致这种疾病的物种和菌株,这是开发多价疫苗的重要信息。本研究的目的是分离与埃塞俄比亚中部选定地区的绵羊肺炎病例相关的曼海姆菌,确定其在该研究地区流行的细菌的作用和菌株/基因型。

结果

对从 76 例绵羊肺炎病例中采集的鼻拭子样本进行的细菌学分析表明,从其中 26 例中分离出曼海姆菌,从 2 例中分离出 B.trehalosi。使用曼海姆菌种特异性和曼海姆菌血清型 1 抗原特异性 PCR 检测对分离株进行的进一步分子分析表明,26 株分离株被鉴定为曼海姆菌,其中 21 株为曼海姆菌血清型 1。尽管在曼海姆菌分离株中观察到非特异性产物,但针对多杀性巴氏杆菌荚膜生物合成基因 (capA) 的 PCR 检测未检测到曼海姆菌和 B.trehalosi 分离株。对本研究中包含的曼海姆菌分离株与参考菌株在 PHSSA 和 Rpt2 基因方面的系统发育分析表明,埃塞俄比亚曼海姆菌分离株构成三个不同的基因型,与起源地一致。

结论

研究表明,曼海姆菌通常与埃塞俄比亚中部研究地区的绵羊肺炎病例有关,尽管仍有待确定导致大多数病例的其他剩余病原体。分子特征表明,在研究地区流行的曼海姆菌存在三种基因型,与分离地点一致。研究结果表明,需要进一步广泛开展工作,以确定与绵羊肺炎相关的所有病原体和曼海姆菌的菌株分布,以了解其在全国范围内的分子流行病学,并制定具有成本效益的预防和控制方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bd81/6280500/ff68a02b393f/12866_2018_1338_Fig1_HTML.jpg

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