基于高通量定量PCR与高通量测序技术研究城市湿地公园抗生素抗性基因污染特征

High-throughput qPCR and Amplicon Sequencing as Complementary Methods for Profiling Antibiotic Resistance Genes in Urban Wetland Parks

城市湿地公园是城市湿地保护与利用的重要实践,具有重要的生态系统服务价值.抗生素抗性基因(ARGs)作为一种新污染物,在水体环境中普遍存在,是当前环境研究和公众关注热点.基于高通量定量PCR与16S rRNA基因高通量测序技术,深入研究了厦门城市湿地公园水体环境中抗生素抗性基因的种类、丰度水平和分布特征.结合微生物群落特征和水质理化性质,阐释了城市湿地公园抗生素抗性基因污染的影响因子和驱动机制.结果表明,城市湿地公园水体环境总共发现了217种抗生素抗性基因,丰度水平高达6.48×10^(9)copies·L^(-1),抗性基因呈现出丰度较高、种类多样以及普遍共存的特点,城市湿地公园是抗生素抗性基因分布的重要热区和存储库;Marivivens、NS5_marine_group和Planktomarina等9种微生物是41种抗生素抗性基因的潜在携带者,微生物群落结构对城市湿地公园水体环境抗性基因变化的单独解释量达到了51%,因此微生物群落是城市湿地抗生素抗性基因赋存和演化最重要的驱动力.基于研究结果,暗示着城市湿地公园水体环境抗生素抗性基因具有潜在水生态安全及人类健康风险,应该加强城市湿地公园水体环境微生物污染研究与控制.

Urban wetland parks are an important practice for urban wetland protection and utilization due to the vast ecosystem service value.As emerging contaminants,antibiotic resistance genes(ARGs)are great attractions for environmental research and public concerns.Based on high-throughput qPCR and high-throughput amplicon sequencing techniques,we investigated the occurrence,abundance,and distribution profiles of antibiotic resistance genes in the aquatic environment of Xiamen urban wetland parks(five sites).The influencing factors and driving mechanisms of antibiotic resistance genes were deciphered on the basis of microbial community structure and water quality.Diverse and abundant ARGs were observed and coexisted in urban wet parks.A total of 217 ARGs were detected in the water body of urban wetland parks,with an abundance up to 6.48×10^(9) copies·L^(-1).Urban wetland parks were important hotspots and repositories of the antibiotic resistome.A total of nine bacterial genera,including Marivivens,NS5_marine_group,and Planktomarina,were identified as the potential carriers of diverse resistance genes(41 ARGs).The microbial communities could alone explain 51%of alterations in the antibiotic resistome in the aquatic environment of the urban wetland parks.Therefore,the microbial community was the key driving force for the occurrence and evolution of ARGs in urban wetland parks.Based on the results,with the presence of ARGs and antibiotic resistance bacteria,it is suggested that the water environments of urban wetland parks have potential risks of water ecological security and human health,and it is necessary to further enhance the research and control of microbial contaminants in the aquatic environment of urban wetland parks.