种蛋孵化死胚及孵化厅环境样本中铜绿假单胞菌的分离鉴定及其耐药性分析

Isolation,Identification and Drug Resistance Analysis of Pseudomonas aeruginosa from Dead Embryos and Environment Samples of Hatchery in Breeding Chicken Farms

本研究旨在了解种鸡场中种蛋孵化死胚和周围环境中感染或污染铜绿假单胞菌(Pseudomonas aeruginosa,PA)的比率及分离株的耐药性。从广东省的5个规模化种鸡场采集孵化厅死胚及其周围环境样本进行PA的分离鉴定,采用K-B纸片扩散法分析其抗菌药物敏感性,并通过常规PCR检测对应的耐药基因。结果显示,在采集的679份样本中,分离到共77株(77/679,11.3%)PA,其中2株来源于环境样本(2/24,8.3%),75株来源于死胚(75/655,11.5%)。药敏试验检测显示,耐药率最高的是复方新诺明(100%)和四环素(100%),其次为氯霉素(75.4%),另外第三代头孢噻肟也有检测出耐药株,耐药率达11.7%;多重耐药率高达62.3%;耐药基因qacE△1-sul1(29.9%)和ant(3″)-I(29.9%)的检出率最高,其次为DHA(28.3%),而oprD_(2)基因阳性率为11.7%,提示其对碳青霉烯类抗菌药的耐药性转移风险水平较高。综上,所调查的5个种鸡场的种蛋孵化死胚及其周围环境中均存在不同程度的PA感染或污染,且分离鉴定的菌株表现出较强的耐药性。因此建议该5个种禽场在养殖过程中要加强饲养管理,及时清洁和消毒种蛋、孵化机及其周边环境;另一方面应定期进行药敏试验,合理使用抗菌药物,以尽量减少耐药及多重耐药菌株的出现。

This study aims to investigate the prevalence and drug resistance of Pseudomonas aeruginosa(PA)in breeder farms in Guangdong Province.Samples of dead embryos of breeding chickens and their surrounding environment were collected from different regions of Guangdong Province for the separation and identification of PA.The K-B paper disk diffusion method was used to analyze its antimicrobial sensitivity,and its resistance genes were detected by conventional PCR.The results showed that a total of 77 strains of PA(77/679,11.3%)including 2 strains from environmental samples(2/24,8.3%)and 75 strains(75/655,11.5%)from dead embryos were isolated.The drug susceptibility test showed that the highest drug resistance rate was compound trimethoprim(100%)and tetracycline(100%),followed by chloramphenicol(75.4%).In addition,drug-resistant strains were also detected in the third-generation cefotaxime,the drug resistance rate reached 9.3%;It is worth noting that the multi-drug resistance rate is as high as 62.3%;the resistance genes qacE△1-sul1(29.9%)and ant(3″)-I(29.9%)have the highest detection rate,followed by DHA(28.3%),and the positive rate of oprD_(2) gene was 11.7%,suggesting that its resistance to carbapenem antimicrobial drugs has a high level of transfer risk.In summary,there were varying degrees of PA infection or pollution in the five breeder farms and their surroundings in Guangdong Province,and the isolated and identified strains showed strong drug resistance.Therefore,it is suggested that the five poultry farms should strengthen the breeding management during the breeding process,clean and disinfect the breeding eggs.On the other hand,drug susceptibility tests should be conducted regularly,and antibiotics should be used rationally to minimize the emergence of drug-resistant and multi-drug resistant strains.