圈养林麝铜绿假单胞菌的分离鉴定及耐药性和病原特性分析

Isolation, identification and drug-resistance, pathogeny characteristics of Pseudomonas aeruginosa from captive forest musk deer

摘要:

【背景】铜绿假单胞菌感染所致的化脓性疾病是困扰林麝驯养的重要因素，是一类较难防治的细菌性疾病，目前尚无疫苗可用来预防该病。【目的】研究林麝源铜绿假单胞菌的感染现状和分子流行病学规律。【方法】对2014年10月至2015年10月四川宝兴和陕西镇坪两个林麝养殖中心发病林麝中铜绿假单胞菌进行分离鉴定，并对其耐药情况进行分析，利用脉冲场凝胶电泳(PFGE)分型技术研究分离菌的PFGE指纹图谱，探究其流行病学趋势，并对部分菌株的致病性进行分析。【结果】分离得到60株铜绿假单胞菌，其中34株来自镇坪，26株来自宝兴。耐药结果表明：60株林麝源铜绿假单胞菌对17种抗菌药物呈现不同程度耐药性，不同地区和不同样本源间分离的铜绿假单胞菌对17种抗菌药物的耐药性总体趋于一致，多重耐药情况严重，以5耐、6耐为主。分型结果显示：60株铜绿假单胞菌PFGE图谱的相似性为49.1%?100%。经聚类分析得到A?O共15种基因簇，其中优势基因簇为C、E、G、J。致病性结果表明，流行菌株的致病力强弱依次为：动物源菌株>环境源菌株，且主要流行菌株(基因簇E、F、J)的致病力大于其余菌群。【结论】铜绿假单胞菌在两地区具有水平传播的途径，本研究可为跨地区林麝化脓性炎症的防控提供理论依据。

Abstract:

[Background] The suppurative disease, a bacterial infection caused by Pseudomonas aeruginosa and relatively difficult to prevent, has been a difficult problem in the domestication and breeding of forest musk deer. Currently, no vaccine is available. [Objective] To study the status of infection and molecular epidemiology of P. aeruginosa. [Methods] P. aeruginosa strains were isolated from two forest musk breeding centers in Zhenping County, Shaanxi Province and Baoxing County, Sichuan Province from October 2014 to October 2015, as well as their drug resistance was tested by traditional bacteriological methods. The fingerprint data bank of all strains was established by pulsed field gel electrophoresis (PFGE) typing, PFGE data were analyzed by BioNumerice software to reveal the prevalence pattern, and the pathogenicity of some isolates was analyzed. [Results] In total 60 P. aeruginosa strains were isolated, of which 34 were isolated from Zhenping, and 26 were isolated from Baoxing. The isolated P. aeruginosa strains were resistant to 17 antimicrobial agents at different levels, and the isolates from different area or different samples exhibited similar resistance phenotype. Multi-resistance was observed among the P. aeruginosa isolates, most of them exhibited multidrug-resistance to 5 and 6 antimicrobial agents. PFGE spectrum similarity of the 60 strains was between 49.1% and 100%, and A to O 15 PFGE types were obtained through cluster analysis, C, E, G, J were the dominant types. The virulence experiment on mice (LD50) suggested that the strains from animals were far more virulent than those isolated from the environmental, and the dominant bacterial type (E, F, J gene type) were more virulent than other strains. [Conclusion] There was horizontal transmission path with P. aeruginosa in different areas. The data provide a basis for prevention and treatment of suppurative inflammation in the captive musk deer.