抗生素作为禽畜疾病预防及治疗药物、生长促进剂、饲料添加剂等被广泛应用于养殖业并进入环境,造成了环境中抗性耐药菌和抗性基因(Antibiotic Resistance Genes,ARGs)的日益增加,从而影响人体健康和生态环境。因此,ARGs作为一种新型环境污染物已经引起了人们的广泛关注。本工作以珠海某些养殖场及周边土壤中的微生物及其ARGs为主要研究对象,采用聚合酶链反应(PCR)研究了养殖场土壤细菌DNA中9种四环素抗性基因存在和污染水平。结果表明,所研究的9种四环素抗性基因中除tetD之外,其他8种抗性基因(tetA、tetB、tetC、taG、tetL、tetO、tetQ和tetX)均可检出,其中tetG的样本检出率达到94%;另外,对其中常见的4种ARGs(tetA、tetC、tetG和tetX)的荧光实时定量PCR定量结果显示,在这4种基因中,以tetC在样品中的丰度最高,tetG次之。在16个土壤采样点中发现,tetC和tetG的copies/gDNA达到10^8量级均占50%,其他基因的copies/gDNA最低也能达到10^2~10^5之间,与北京某一养猪场周边土壤中抗性基因含量相当,有些甚至更为严重。
Use of antibiotics at subtherapeutic concentrations for agricultural applications has been believed to be an important factor in the proliferation of bacteria with antibiotic resistance genes i ARGs). In this work, total nine tetracycline ARGs (tet-ARGs) including tetA, tetB, tetC, tetD, tetG, tetL, tetO, tetQ and tetX) were studied in sixteen soil samples from livestock using qualitative PCR SYBR Green I real-time PCR methods. The results showed that eight of tet-ARGs except tetD could be detected in most of samples and the detection percentages ranged from 6% (tetB) to 94% (tetC). The quantitative results for four tetgenes (tetA, tetC, tetG and tetX) showed that 50% samples had the highest tetC abundance ( 10^8 ) and the others were in range of 10^2 to 10^5. As a result, the spread occurrences of high levels of tet-genes in livestock soil should not be overlooked.