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动物源金黄色葡萄球菌生物被膜形成能力与分子分型关系研究
引用本文:唐子云,胡健欣,陈进,陆毅兴,孔伶俐,刁露,张发福,熊文广,曾振灵. 动物源金黄色葡萄球菌生物被膜形成能力与分子分型关系研究[J]. 中国农业科学, 2022, 55(3): 602-612. DOI: 10.3864/j.issn.0578-1752.2022.03.014
作者姓名:唐子云  胡健欣  陈进  陆毅兴  孔伶俐  刁露  张发福  熊文广  曾振灵
作者单位:华南农业大学兽医学院/广东省兽药研制与安全评价重点实验室/岭南现代农业科学与技术广东省实验室/国家兽医微生物耐药性风险评估实验室,广州 510642
基金项目:“十三五”国家重点研发计划(2018YFD0500300);
摘    要:[目的]调查产生生物被膜的金黄色葡萄球菌流行病学特征,探究生物被膜形成能力与分子型之间的相关性,为治疗动物金黄色葡萄球菌感染提供理论依据.[方法]以广东省兽药研制与安全评价重点实验室保存的98株金黄色葡萄球菌为研究对象,用结晶紫半定量法测定所有菌株生物被膜形成能力,将产膜菌株进行22种常见抗菌药的最小抑菌浓度测定,通过...

关 键 词:金黄色葡萄球菌  生物被膜  耐药性  分子分型  毒力基因
收稿时间:2020-12-10

Relationship Between Biofilm Formation and Molecular Typing of Staphylococcus aureus from Animal Origin
TANG ZiYun,HU JianXin,CHEN Jin,LU YiXing,KONG LingLi,DIAO Lu,ZHANG FaFu,XIONG WenGuang,ZENG ZhenLing. Relationship Between Biofilm Formation and Molecular Typing of Staphylococcus aureus from Animal Origin[J]. Scientia Agricultura Sinica, 2022, 55(3): 602-612. DOI: 10.3864/j.issn.0578-1752.2022.03.014
Authors:TANG ZiYun  HU JianXin  CHEN Jin  LU YiXing  KONG LingLi  DIAO Lu  ZHANG FaFu  XIONG WenGuang  ZENG ZhenLing
Affiliation:College of Veterinary Medicine, South China Agricultural University/Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation/Guangdong Laboratory for Lingnan Modern Agriculture/National Risk Assessment Laboratory for Antimicrobial Resistance of Animal Original Bacteria, Guangzhou 510642
Abstract:【Objective】 The aim of this study was to investigate the epidemiological characteristics of Staphylococcus aureus (S. aureus) in the biofilm producing strains and to explore the correlation between biofilm forming ability and molecular typing, so as to provide the theoretical basis for the treatment of S. aureus infection. 【Method】 The biofilm producing ability of all strains of S. aureus was determined by crystal violet semi-quantitative method. The minimum inhibitory concentrations of 22 common antibiotics were determined by the membrane producing strains. Molecular typing was conducted by three common typing methods of S. aureus, including spa typing, MLST typing and PFGE typing, and the correlation between membrane production capacity and molecular typing was analyzed. Finally, whole genome sequencing technology was used to analyze the antibiotics resistance gene and virulence genes in the biofilm producing strains. 【Result】 The semi-quantitative results of crystalline violet showed that a total of 23 strains (23.47%) of 98 S. aureus strains were able to produce biofilm, including 22 strains (25.29%, 22/87) from cow milk source, 14 strains (60.87%, 14/22) from Zhejiang dairy farms, 8 strains (39.13%, 8/22) from Fujian dairy farms, and 1 strain from pig source (9.10%, 1/11) from Guangdong slaughterhouse, indicating that the film-producing potential of S. aureus from cow's milk source was higher than that of pig source, 22 strains (95.65%) of which were from cow's milk source and 1 strain (4.35%) was from pig source. The film-producing ability was classified into strong, medium and weak, and among the 23 film-producing strains, 2 strains (8.70%, 2/23) were strong film-producing strains, 9 strains (39.13%, 9/23) were medium, and 12 strains (52.17%, 12/23) were weak. The results of the drug sensitivity test showed that the bovine milk-derived membrane- producing strains were sensitive to all the tested antibacterial drugs, while the pig-derived membrane-producing strains showed resistance to 13 antibacterial drugs, including penicillin, amoxicillin, ceftiofur, cefoxitin, enrofloxacin, ciprofloxacin, clindamycin, doxycycline, erythromycin, flupenthixol, cotrimoxazole, tiamulin, and tilmicosin. The spa typing results showed that 98 strains of S. aureus obtained 8 spa types, and 23 strains of film-producing S. aureus accounted for 3 of them: 1 strain t2922 from porcine origin in Guangdong, 14 strains t2119 from Zhejiang cow milk source, and 8 strains t189 from Fujian cow milk source. MLST typing results showed that 98 strains were classified into 9 ST types, of which 6 ST types did not have the ability to produce biofilm, namely ST398, ST522, ST705, ST1651, ST479 and ST151, and only 3 strains of ST type had biofilm production ability, namely ST9, ST7 and ST188. It was found that the molecular types of strong film-producing strains were mainly ST7-t2119, the medium film-producing strains were mainly ST7-t2119 and ST188-t189, and the weak film-producing strains were ST9-t2922, ST7-t2119 and ST188-t189. The ST type of weak film-producing strains could be well distinguished from the medium and strong film-producing strains, and only the specific ST type of S. aureus had the ability to produce biofilm. 23 film-producing strains PFGE typing all successful PFGE typing results showed that the results show that each strain of film-producing bacteria in Guangdong, Fujian and Zhejiang provinces were divided into 3 PFGE types, and there were geographical distribution characteristics of PFGE types; the strains isolating from the same region had clonal transmission, and the strains in the province were clonal to each other, but there were significant differences in biofilm production ability between clones.; the whole genome sequencing results showed that the drug resistance genes and virulence genes in the film-producing strains were diverse according to the molecular type. 【Conclusion】 S. aureus from different sources had different potential to produce biofilm and all carried different film-producing genes. The film-producing potential of S. aureus from bovine milk source was much higher than that of porcine source, and all carrid different film-producing genes. Whether strains could produce film or not may be strongly correlated with ST type, and the specific ST types, such as ST9, ST7 and ST188, were more likely to produce biofilm; however, at the same time, the strains with the same molecular type had different abilities to produce biofilm.
Keywords:Staphylococcus aureus  biofilm  antibiotics resistance gene  molecular typing  virulence genes  
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