多种动物源A型产气荚膜梭菌的分离鉴定及生物学特性

Isolation,Identification and Biological Characterization of Clostridium perfringens type A from Multiple Animal Origins

【目的】研究猪源及其它动物源A型产气荚膜梭菌(CpA)的β2毒素基因和相关生物学特性。【方法】从南昌周边地区随机采集猪、牛、羊、鸡、犬、鱼的新鲜粪便样品,采用4%D-环丝氨酸FTG梭菌选择培养基从各样品中分离纯化产气荚膜梭菌,并对分离菌株进行培养特性观察、革兰氏染色镜检、生化鉴定和16S rDNA分子鉴定;再通过多重PCR技术进行分子分型,对分离到的A型产气荚膜梭菌(CpA)进行β2毒素基因检测和溶血试验。【结果】所分离到的CpA中,除犬源CpA,其它动物源分离株均携带β2毒素基因;不同动物源的CpA溶血时间和溶血强度不一样:羊源CpA在血平板上4 h能观察到明显的溶血,溶血圈直径达到30 mm;猪源CpA 6 h才能观察到明显的溶血,溶血圈直径约为14 mm。【结论】为了解CpA致病机理,解析其在不同动物致病强弱和地区特异性提供参考依据。

[Objective]To study theβ2 toxin gene and biological characterization ofClostridium perfringens type A(CpA)from pig and otheranimals.[Method]The samples of fresh feces from pigs,cattle,sheep,chickens,dogs and fish were collected randomly in the surrounding area of Nanchang.Clostridium perfringens was isolated from each sample by the FTG selective medium containing 4%D-Cycloserine.Observations on the isolated strains were conducted,including culture characteristics,Gram staining microscopic examination,biochemical identification,16s rDNA molecular identification and hemolysis test.The molecular typing was carried out andβ2 toxin of CpA isolated from various animal isolates were detected by multiplex PCR assay.[Result]The results showed that all the isolations but canine origin CpA carriedβ2 toxin gene.The hemolysis time and intensity of CpA from various animals varied.The hemolysis in blood plate of CpA from sheep was observed about two hours earlier than that of the porcine CpA,the diameters of the hemolysis circles were about 14 mm and 30 mm.[Conclusion]This study provides a theoretical basis for understanding of the pathogenic mechanismof CpA and analysis of its pathogenic and regional specificity in different animals.