猪源葡萄球菌的分离鉴定及耐药性分析

Isolation,Identification and Drug Resistance of Staphylococcus from Swine

为明确从福建某规模猪场分离到的病原菌及其生物学特性、致病性与耐药等特征,为疫病的防治提供参考依据。对分离菌进行纯化,采用革兰氏染色、PCR鉴定、药敏试验、消毒剂消杀试验、耐药基因检测及动物攻毒试验等,对分离菌进行生物学特性、耐药等相关检测。结果显示,分离菌为猪源葡萄球菌。该菌对30种常用抗菌药物中的苯唑西林、诺氟沙星、头孢类等14种药物高度敏感,对阿米卡星、庆大霉素、四环素、万古霉素、氟苯尼考5种药物中度敏感,对氨苄西林、哌拉西林、卡那霉素等11种药物不敏感,检测cfr、tetM、floR、mcr-1、bla NDM-1、optrA 6种耐药基因均为阴性;该菌对26种中药中的单宁酸高度敏感,对五味子、五信子多酚中度敏感,对叶下珠、厚朴、虎杖低度敏感,对半边莲、铁苋菜、艾叶、重楼等20种中药不敏感;在8种消毒剂中,84消毒液对该分离菌的消杀效果差,新洁尔灭、月苄三甲氯铵、复合酚3种消毒剂对该分离菌消杀效果好;动物攻毒试验表明,该菌对25日龄健康断奶仔猪具有强致病性,能导致猪出现典型的葡萄球菌病临床症状和病理特征,攻菌猪组织病理可见脾脏红髓淤血、肺泡腔内见红细胞渗出及少量粉染的渗出物。研究结果可为在养殖饲料端禁止添加抗生素的背景下,为猪葡萄球菌病的防治提供理论参考和技术支撑。

To clarify the pathogenic bacteria isolated from a large-scale pig farm in Fujian,their biological characteristics,pathogenicity,and resistance to Chinese and western medicines were conducted to provide reference basis for the prevention and control of diseases.In the test,the isolated bacteria were purified,and the biological characteristics,resistance to Chinese and western medicines and other related tests of the isolated bacteria were carried out by Gram stain,PCR identification,drug sensitivity test,disinfectant disinfection and sterilization test,drug resistance gene test and animal challenge test.The results showed that the isolated bacteria were Staphylococcus.The bacterium is highly sensitive to 14 of the 30 commonly used western antimicrobials,including oxacillin,norfloxacin and cephalosporins.It is moderately sensitive to 5 drugs,including amikacin,gentamicin,tetracycline,vancomycin and florfenicol.It is not sensitive to 11 drugs,including ampicillin,piperacillin and kanamycin.The detection of six resistance genes,including cfr,tetM,floR,mcr-1,blaNDM-1 and optrA,is negative;The bacterium is highly sensitive to tannic acid in 26 kinds of traditional Chinese medicines,moderately sensitive to Schisandra chinensis and Wuxinzi polyphenols,low sensitive to Yexiazhu,Magnolia officinalis,Polygonum cuspidatum,and insensitive to 20 kinds of traditional Chinese medicines,such as Lobelia chinensis lobata,iron amaranth,wormwood leaf,Paris polyphylla;Among the eight disinfectants,84 disinfectant had poor disinfection and sterilization effect on the isolated bacteria,while three disinfectants,namely bromogeramine,trimethylammonium chloride and compound phenol,had good disinfection and sterilization effect on the isolated bacteria;The animal challenge test showed that the bacterium was highly pathogenic to 25 day old healthy weaned piglets,and could lead to typical clinical symptoms and pathological characteristics of staphylococcosis in pigs.The histopathology showed congestion of the spleen red pulp,exudation of red blood cells in the alveolar cavity,and a small amount of powder stained exudate.The research aims to isolate pig derived Staphylococcus aureus and clarify its pathogenicity and drug resistance in the context of prohibiting the addition of antibiotics in the feed,providing theoretical reference and technical support for the prevention and treatment of pig staphylococcosis.