养殖场废水中磺胺类和四环素抗生素及其抗性基因的定量检测  

Quantitative detection of sulfonamides and tetracycline antibiotics and resistance genes in sewage farms

作　　者：冀秀玲[1] 刘芳[1] 沈群辉[1] 刘扬[1] 

机构地区：[1]东华大学环境学院环境科学系,上海201620

出　　处：《生态环境学报》2011年第5期927-933,共7页Ecology and Environmental Sciences

基　　金：中央高校基本科研业务经费(9D11306);上海市重点学科开放基金项目(B604)

摘　　要：抗生素滥用及其诱导产生的抗生素抗性基因（antibiotic resistance genes,ARGs）污染已经引起人们的广泛关注。选取上海市某地养殖场作为研究对象,采用高效液相色谱-质谱法和实时荧光定量PCR法,对养殖场污水及附近农田灌溉渠河水中5种四环素及磺胺类抗生素,8种对应的ARGs的含量、特征及相关性进行了研究。研究结果显示,在采集的水样中均含有待检测的5种抗生素,养殖污水中抗生素含量高于农田灌溉河水,各样本中四环素类抗生素含量均略高于磺胺类抗生素,2种四环素抗生素总量为294.0~376.1μg﹒L-1,3种磺胺类抗生素总量为197.7~323.0μg﹒L-1。养殖场污水样本中8种ARGs均有检出,磺胺类抗性基因中sul（A）含量最高,绝对拷贝数为108.4108~1010.3728;四环素类抗性基因中tet（W）含量最高,绝对拷贝数为106.18805~107.8874。农田灌溉渠河水中除tet B（P）外,其它7种ARGs均有检出。样本中ARGs相对表达量总体呈现磺胺类ARGs高于四环素类ARGs的特点。抗生素浓度与ARGs相对表达量的Pearson相关性分析显示,样本中sul（Ⅲ）与磺胺类抗生素浓度存在较明显的正相关性,但其它几种ARGs与抗生素则未见或存在一定负相关性。表明除抗生素外,ARGs在水环境中的丰度可能与ARGs种类及其它环境因子有关。该研究将有助于认识上海地区养殖场废水中抗生素和ARGs污染状况,为进一步开展黄浦江水域抗生素尤其是ARGs的污染研究提供数据基础。The emerging pollution of antibiotics and antibiotic resistance genes （ARGs） has gained great attention around the globe in recent years. In this study, high performance liquid chromatography - mass spectrometry （HPLC-MS） and real-time quantitative PCR （RT-PCR） were used for detecting occurrence of two tetracyclines [tetracycline （TC）, oxtetracycline （OTC）], three sulfonamides [sulfamerazine （SDZ）, sulfamethazine （SDMe）, sulfamethoxazole （SMZ）], and 8 ARGs [sul （Ⅰ）, sul （Ⅱ）, sul （Ⅲ）, sul （A）, tet （M）, tet （O）, tet （W）, tet B（P）] in water samples collected from feedlot wastewater lagoons and irrigation ditch in Shanghai. The detection frequencies of the above 5 antibiotics were 100% in all samples. Antibiotics in feedlot wastewater lagoons are much higher than those in irrigation ditch. The total concentrations of sulfonamides and tetracyclines are 197.7-323.0μg · L^-1 and 294.0-376.1 μg · L^-1 respectively. In the collected samples, both the absolute copies and the relative abundances of sulfonamide ARGs are higher than those of tetracycline ARGs. The highest relative abundance of sulfonamide ARGs is sul （A）, and tet （W） is the highest among tetracycline ARGs, with the absolute copies 10^8.410 8-10^10.372 8 and 10^6.1880 5-10^7.887 4 respectively. All of the ARGs were detected in the two feedlot wastewater samples. 7 ARGs, except for tet B（P）, were detected in the irrigation ditch. In Pearson correlation analysis, almost no consistent positive correlations were found between ARGs and ＂their＂ antibiotics, except sul （III） maintained a significant positive correlation with sulfonamide antibiotics. It indicated that the abundance of ARGs might be affected by other kinds of envi- ronmental factors besides antibiotics residues in aquatic environments. This study will help to know the pollution status of the antibiotics and resistance genes in feedlot wastewater in Shanghai and provide valuable data for conducting further rese

关 键 词：抗生素 抗性基因 高效液相色谱-质谱法 实时荧光定量PCR 污水 养殖场 

分 类 号：X132[环境科学与工程—环境科学]