养猪场空气中抗性基因和条件致病菌污染特征

养殖场空气中含有较高浓度的抗生素抗性基因和条件致病菌,对人畜健康具有潜在威胁.采用中流量TSP采样器在某养猪场的生活区、猪舍内和猪舍外3个地点分别采样24 h和48 h,并采集猪舍内的饲料、粪便和饮水添加剂样品.采用普通PCR检测样品中的3类抗生素抗性基因(大环内酯类、β-内酰胺类、四环素类各3个基因)和7种致病菌/条件致病菌基因(弯曲杆菌属、产气荚膜梭菌、肠球菌属、大肠杆菌、小肠结肠炎耶尔森菌、葡萄球菌属和猪链球菌);选取检出率较高的6种基因,采用荧光定量PCR对其浓度进行测定.结果表明: 空气中大环内酯类抗性基因检出了3个,四环素类检出了2个,肠球菌、大肠杆菌、小肠结肠炎耶尔森菌、葡萄球菌等4种条件致病菌在空气样品中和饮水添加剂中都被检测到.绝大部分目的基因的浓度均在10⁴ copies·m^-3以上,并且猪舍附近浓度远高于生活区;猪场内主要的抗生素抗性基因和条件致病菌的可能来源是猪粪便和饮水添加剂.在养猪场内采样24 h即可满足PCR检测要求;在生活区采样48 h的采样效率高于采样24 h,而在猪舍外和猪舍内采样24 h的效率高于采样48 h.

Sources and pollution characteristics of antibiotic resistance genes and conditional pathogenic bacteria in concentrated swine feeding operation.

Air in concentrated animal feeding operations contains antibiotic resistance genes and airborne pathogens, with potential threat to human and animal health. In this study, air was sampled in the living area, outside, and inside of a fattening pig house in a pig farm for 24 and 48 hours. Feedstuffs, drinking water additives, and feces in the pig house were collected. Three kinds of antibiotic resistance genes (macrolide, β-lactam, and tetracycline) and seven pathogenic microorganisms (Campylobacter, Clostridium perfringens, Enterococcus, Escherichia coli, Yersinia enterocolitica, Staphylococcus spp., and Streptococcus suis) were detected by polymerase chain reaction (PCR). Six genes with high detection rates were selected, with their concentrations being determined by real-time quantitative PCR (qPCR). Results showed that three macrolide and two tetracycline resistance genes were detected in all air samples. Enterococcus, Escherichia coli, Yersinia enterocolitica, and Staphylococcus spp. were detected in air samples and drinking water additive. The concentrations of most target genes were above 10⁴ copies·m^-3. The gene concentrations near the pig house were much higher than those in the living area. Main sources of antibiotic resistance genes and pathogens in the air were pig manure and drinking water additive. Sampling time of 24 h in the pig farm met the requirements for PCR detection. Sampling time of 48 h had a higher sampling efficiency than that of 24 h in the living area of the pig farm, whereas sampling time of 24 h was more appropriate than that of 48 h in high bioaerosol concentration area such as the pig house.