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猪肺炎支原体田间分离株毒力评估

Evaluation of virulence of Mycoplasma hyopneumoniae field isolates.

作者信息

Vicca J, Stakenborg T, Maes D, Butaye P, Peeters J, de Kruif A, Haesebrouck F

机构信息

Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.

出版信息

Vet Microbiol. 2003 Dec 30;97(3-4):177-90. doi: 10.1016/j.vetmic.2003.08.008.

Abstract

The course of enzootic pneumonia, caused by Mycoplasma hyopneumoniae, is strongly influenced by management and housing conditions. Other factors, including differences in virulence between M. hyopneumoniae strains, may also be involved. The aim of this study was to evaluate the virulence of six M. hyopneumoniae field isolates and link it to genetic differences as determined by randomly amplified polymorphic DNA (RAPD) analysis. Ninety, conventional M. hyopneumoniae-free piglets were inoculated intratracheally with the field isolates, a virulent reference strain or sterile culture medium. Animals were examined daily for the presence of disease signs and a respiratory disease score (RDS) was assessed per pig. Twenty-eight days post infection, pigs were euthanized, blood sampled and a lung lesion score was given. Lung samples were processed for histopathology, immunofluorescence testing for M. hyopneumoniae and isolation of M. hyopneumoniae. RAPD analysis was performed on all M. hyopneumoniae strains. Significant differences between isolates were found for the RDS, lung lesion score, histopathology, immunofluorescence and serology. Based on the results of the different parameters, isolates were divided into three "virulence" groups: low, moderately and highly virulent strains. Typically, a 5000 bp RAPD fragment was associated with the highly and moderately virulent strains whereas it was absent in low virulent strains. It was concluded that high variation in virulence exists between M. hyopneumoniae strains isolated from different swine herds. Further studies are required to determine whether the 5000 bp fragment obtained in the RAPD analysis can be used as a virulence marker.

摘要

由猪肺炎支原体引起的地方性肺炎病程,受到管理和饲养条件的强烈影响。其他因素,包括猪肺炎支原体菌株之间毒力的差异,也可能起作用。本研究的目的是评估六株猪肺炎支原体田间分离株的毒力,并将其与通过随机扩增多态性DNA(RAPD)分析确定的基因差异联系起来。九十头常规无猪肺炎支原体的仔猪经气管内接种田间分离株、一株强毒参考菌株或无菌培养基。每天检查动物是否有疾病体征,并对每头猪评估呼吸疾病评分(RDS)。感染后28天,对猪实施安乐死,采集血液样本并给出肺损伤评分。对肺样本进行组织病理学处理、猪肺炎支原体免疫荧光检测以及猪肺炎支原体分离。对所有猪肺炎支原体菌株进行RAPD分析。在RDS、肺损伤评分、组织病理学、免疫荧光和血清学方面,分离株之间存在显著差异。根据不同参数的结果,将分离株分为三个“毒力”组:低毒、中等毒和高毒菌株。通常,一个5000 bp的RAPD片段与高毒和中等毒菌株相关,而低毒菌株中不存在该片段。得出的结论是,从不同猪群分离的猪肺炎支原体菌株之间存在毒力高度差异。需要进一步研究以确定在RAPD分析中获得的5000 bp片段是否可作为毒力标记物。

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