鹅源产气荚膜梭菌的分离鉴定与药物敏感性试验

Isolation,identification of goose source Clostridium perfringens and in vitro antibacterial effect with Chinese herbal medicine

对2020年7月至2021年6月临床症状与病理剖检初步诊断为鹅坏死性肠炎疑似病例肠道内容物和肝脏进行细菌分离并纯化,获得52株菌。分离菌株经培养特性、生化试验、种特异性PCR鉴定,确定52株菌均为产气荚膜梭菌;采用PCR对分离株毒素基因鉴定,结果显示分离株均为毒素型A型,其中cpβ_(2)基因的检出率为61.5%,未检测到肠毒素cpe基因和netB毒素基因。药敏试验结果显示,分离株对头孢噻肟(100%)、头孢噻呋(100%)、氟苯尼考(100%)和多西环素(94%)表现为高度敏感,其次为恩诺沙星(88%)和氧氟沙星(88%);而耐药率最高的药物为磺胺类药物,其中磺胺甲恶唑耐药率为100%,其次是林可胺类药物,林可霉素与克林霉素耐药率分别为58%和46%;中药体外抑菌试验结果显示,单味中药黄连、黄柏、黄芩和复方中药乌梅散、白头翁汤、郁金散对产气荚膜梭菌均有一定的抑菌作用,其中黄连的MIC值为1.57 g/L,远低于其他试验中药的MIC值,具有较强的抑菌作用。本试验为广东省主要鹅养殖区域鹅产气荚膜梭菌病的有效防治提供了参考依据。

In order to understand the prevalence and the main biological characteristics of goose source Clostridium perfringens(CP)wihch was considered to be the pathogen inducing goose necrotizing enteritis(NE)in Guangdong Province,by measuring their culture characteristics,biochemical tests and PCR identification.52 CP strains were isolated and identificated from the intestinal mucosa and livers of geese showing significant NE clinical symptoms and pathological autopsy from 2020-2021.Furtherly,PCR metod was employed for identifying the toxin genes of the isolates,suggesting that the isolates were toxin type A,The detection rate of cpβ2 gene was 61.5%,and no enterotoxin cpe gene and netB toxin gene were detected.Moreover,the drug sensitivity test showed that isolates were highly sensitive to cefotaxime(100%),cefotaxime(100%),florfenicol(100%)and doxycycline(94%),followed by enrofloxacin and ofloxacin(88%).The sulfonamides,lincomycin and clindamycin were the first three antibiotics resisted by CP strains with resistance rates 100%,58%and 46%,respectively.Furthermore,under single dosage form,the CP strains were sensitive to the Coptis chinensis,Phellodendron amurense,Scutellaria baicalensis in vitro while the compound form,such as Wumei powder,Pulsatilla chinensis and tulip showed antibiotic effects against CPs.Noteworthy,among the above compound forms,Coptis chinensis had the lowest MIC 1.57 g/L.This experiment provided a reference for the epidemic status and Prevention of goose necrotizing enteritis in the main goose breeding areas of Guangdong Province.