摘要
为了探索宁夏平原地区农田土壤中兽药残留和抗生素抗性基因(ARGs)分布差异,选取位于宁夏平原的15个区域,采集农田土壤、堆肥样本,采用超高效液相色谱质谱联用技术检测了这些样本中48种兽药残留情况,同时采用磁珠法提取纯化基因组脱氧核糖核酸(DNA),利用宏基因组二代测序技术对提取的基因组DNA进行测序,与现有的数据库进行比对,分析了研究区主要土壤微生物群落的结构特征以及ARGs的分布特征。结果表明,在土壤及堆肥样本中检测出7种兽药,其中甲氧苄啶的检出率为33.3%。变形菌门(Proteobacteria)和拟杆菌门(Bacteroidetes)两个门类微生物的相对丰度总量占各采样点的7.6%~79.6%。在宁夏平原南部地区厚壁菌门(Firmicutes)相对丰度较高,为4.1%~29.8%,个别地区堆肥样本中放线菌门(Actinobacteria)的相对丰度高达48.65%。土壤及堆肥样本中共检出99种ARGs,抗性基因种类较为相似,相对丰度较高的ARGs为macB、rpoB2、evgS等基因。各采样点样本中转座子(transposons)、自转移广宿主质粒(self-transmissible broad host range plasmids)以及插入序列(insertion sequences)三大类可移动基因元件(MGEs)的相对丰度占各样本比对出的MGEs总量的86.3%~97.0%。研究结果还表明,变形菌门、拟杆菌门是研究区土壤环境中的优势菌门,微生物的群落分布区域化明显,呈现北部、中部、南部采样区域各异的趋势。四环素类、糖肽类、大环内酯类、肽类是研究区土壤环境中最主要的4大类ARGs。转座子、自转移广宿主质粒、插入序列是土壤环境中主要含有的三大类MGEs。
In order to explore the occurrence and distribution of antibiotic resistance genes and veterinary drug residues in the cropland of Ningxia Plain region,the study focused on samples collected from 15 areas around the Ningxia Plain,inlcuding cropland soil samples and manure composts samples.A Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry(UPLC-MS/MS)method was used to detect residual situation of 48 veterinary drugs,and genomic DNA extraction and purification was conducted by magnetic bead method.Extracted genomic DNA were sequenced by metagenomic next-generation sequencing,and microbial community characteristics and antibiotic resistance genes distribution were identified by comparison with existing databases.Results showed that,seven veterinary drugs were detected in samples,and the detection rate of trimethoprim was 33.3%.The combined relative abundance of Proteobacteria and Bacteroidetes accounted for 7.6%to 79.6%at each sampling site.In the southern part of Ningxia Plain,the relative abundance of Firmicutes was higher,ranging from 4.1%to 29.8%.The relative abundance of Actinobacteria in compost samples from the individual areas was as high as 48.65%.A total of 99 antibiotic resistance genes(ARGs)were detected in the cropland soil and manure composts samples.The ARGs with higher relative abundance were macB,rpoB2,evgS,ect.The relative abundance of three major classes of mobile genetic elements(MGEs),including transposons,self-transmissible broad host range plasmids,and insertion sequences,ranged from 86.3%to 97.0%of the total MGEs identified in each sample.The research findings also indicated that Proteobacteria and Bacteroidetes were the dominant bacterial phyla in the soil environment of the study area,with distinct regional patterns in microbial community distribution,showing varying trends among northern,central,and southern sampling regions.Tetracyclines,glycopeptides,macrolides,and peptides were the four most predominant classes of ARGs in the soil environment of the study area.Transposons,self-transmissible broad host range plasmids,and insertion sequences were the three major types of MGEs primarily present in the soil environment.
作者
朱捷
李瑞雪
许立慧
王泽岚
何微
ZHU Jie;LI Ruixue;XU Lihui;WANG Zelan;HE Wei(Ningxia Food Testing and Research Institute,Key Laboratory of Quality and Safety of Wolfberry and Wine for State Administration for Market Regulation,Yinchuan Ningxia 750002;College of Veterinary Medicine,Nanjing Agricultural University,Nanjing Jiangsu 210095)
出处
《环境污染与防治》
北大核心
2025年第6期103-110,I0006,共9页
Environmental Pollution & Control
基金
宁夏回族自治区自然科学基金资助项目(No.2022AAC03718)。
