抗生素的环境残留、生态毒性及抗性基因污染

抗生素的环境污染与生态毒害问题近年来引起了极大的关注.在总结国内外相关研究基础上,对环境中几种典型抗生素(四环素、土霉素、氯四环素和磺胺类等)的污染源以及在水和土壤环境中的残留与污染水平进行了分析;对抗生素的污染生态毒性最新研究进展给予了评述;对抗生素抗性基因在环境中可能的暴露途径进行了探讨,指出应将抗生素抗性基因作为一类新的环境污染物.鉴于我国抗生素污染的严峻事实,建议应从国家层面上尽快开展有关抗生素环境污染和生态毒害机理的系统研究.     

基于宏基因组学解析不同污水处理系统的耐药基因组分布特征和传播机制

污水处理厂是向水环境中传播抗生素抗性基因(antibiotic resistance genes, ARGs)的热点。与城镇污水相比,工业园区废水成分复杂、污染物浓度高,更有利于ARGs的增殖和扩散。为探究不同类型废水环境的ARGs组成特征和潜在的传播风险,采用宏基因组学技术分别对城镇生活污水处理系统(W1-SD)、工业园区废水处理系统(W1-SI)和2个城镇综合污水处理系统(W2-LH1和W2-LH2)进行取样调查。结果显示,多重耐药类、磺胺类、氨基糖苷类和杆菌肽类抗性基因是废水环境中的主要耐药类型,Ⅰ型整合子、转座酶基因等可移动遗传元件(MGEs)对sul1、aadA和ereA等基因亚型的增殖扩散发挥了关键作用,通过序列分型发现质粒型ARGs的相对丰度更高,尤其是在进水样品中,氨基糖苷类和磺胺类等抗性基因是主要的质粒型ARGs;污水处理过程削减了ARGs多样性,且经过二次沉淀工艺,ARGs丰度均明显降低,但在W1-SI和W2-LH2中,后续的深度处理工艺又使ARGs丰度升高;与城镇污水处理系统相比,W1-SI的ARGs组成更为稳定,最终排水中富集了较高丰度的质粒型ARGs,同时识别...     

CTX-M型产ESBLs耐药基因在城市典型河流中的生态分布

为了解流经我国城市的典型河流中CTX-M型耐药基因的分布情况及基因变异的多态性现状,进而为科学预防和避免抗生素耐药性的产生和传播提供生态学理论支持,采集我国东部6条典型河流流经城市区段的水样,采用宏基因组学技术与限制性酶切片段多态性分析(RFLP)技术相结合对质粒宏基因组PCR产物进行了分析,并将数据进行生态学统计处理.结果显示CTX-M三种亚型的基因群落在调查中的6条河流均有分布,且都保持了较高的均匀度(E≥0.9298),亚型1中分布于长江水体的基因群落具有明显高于其他河流和平均值的多样性(H’=2.4774)和物种优势度(D=0.9107),而亚型2、3的多样性和优势度最高值则均出现在黄河水体中;松花江与海河之间的群落相似性最高,达到0.806 2,而黄河与珠江基因群落之间的相异性则达到了0.580 0.说明在调查中采样的6条河流中均分布着结构稳定且基因优势突出的CTX-M型耐药基因,已对水体生态安全构成了隐患,需要结合进一步研究对其进行防治,以避免耐药基因以质粒为媒介向其他微生物传播造成抗生素耐药性扩散.     

北江河水中抗生素抗性基因污染初步研究（Preliminary Studies on the Pollution Levels of Antibiotic Resistance Genes in the Water of Beijiang River, South China）

环境中细菌的耐药性,尤其是对抗生素的耐药性已成为影响生态环境的重要因素.论文采用抗生素抗性平板法调查了北江河水中四环素、红霉素及磺胺类这3类抗生素耐药性细菌的存在,采用定性PCR及荧光定量PCR方法分别研究了该水域sul1和sul2这2种磺胺类抗生素抗性基因(ARGs)的存在和含量水平.结果表明,所采集北江水域的9个样品中有5个对四环素有耐药性,7个对红霉素有耐药性,8个对磺胺二甲嘧啶有耐药性;定性PCR实验并经基因测序结果证实,5个样品含有sul1,4个样品含有sul2.进一步的PCR定量分析结果显示,7个样品中均检出sul1和sul2磺胺抗性基因,它们与内对照基因16S-rRNA表达量比值分别在10-2.56~10-0.52及10-3.25~10-1.24范围内,该结果显著高于美国科罗拉多州北部河流的研究结果.此外,数据分析也发现,sul1和sul2磺胺抗性基因的含量水平与该区域水中磺胺含量分布具有一定的相关性,表明外源性抗生素对河流的污染是诱导抗性基因的重要因素.     

MBR工艺处理螺旋霉素制药废水过程中抗生素耐药菌与抗性基因的研究

抗生素废水含有大量的抗生素耐药菌(antibiotic resistant bacteria,ARB)与抗性基因(antibiotic resistance genes,ARGs),处理排放后可能增强受纳环境的微生物抗性,因此有必要深入研究抗生素废水处理过程中ARB与ARGs的削减效果及其影响因素。本研究采用膜生物反应器(membrane bioreactor,MBR)工艺处理螺旋霉素制药废水,考察了不同水力停留时间(hydraulic retention time,HRT)对ARB与ARGs削减效果的影响。结果表明,虽然在HRT=30 h时MBR对COD与氨氮的去除率略高于HRT=40 h,但HRT=40 h时,不仅总异养菌与肠球菌的去除效果更佳,出水肠球菌耐药率及携带的抗性基因检出率也更低,而且MBR对废水中erm B、erm F、erm X、mef A、ere A、mph B和转移元件ISCR 1、Tn 916/1545相对丰度的削减效果更好。这表明长HRT更有利于MBR工艺削减螺旋霉素废水的耐药菌与抗性基因。     

我国养猪业废弃物中四环素类、磺胺类抗生素及相关抗性基因污染研究进展

我国是世界养猪第一大国,生猪饲养量和猪肉产量均位居世界第一。养猪业每年所产生的粪便、废水中含有大量畜用抗生素及其代谢产物,使养猪业废弃物成为环境中重要的抗生素污染源之一,随之产生的抗性基因污染及传播问题也不容忽视。本文结合近年来国内外的研究数据,对我国养猪业废弃物中四环素类、磺胺类抗生素及其相关抗性基因的检测方法、污染状况及影响抗性基因传播的因素进行了分析,并基于控制我国养猪行业抗生素及抗性基因污染的目的,提出了今后的研究重点。     

Degradation of extracellular genomic,plasmid DNA and specific antibiotic resistance genes by chlorination

Extracellular DNA structure damaged by chlorination was characterized. Integrity of extracellular ARG genetic information after chlorination was determined. Typical chlorine doses will likely effectively diminish extracellular DNA and ARGs. Plasmid DNA/ARGs were less readily broken down than genomic DNA. The Bioanalyzer methodology effectively documented damage incurred to DNA. There is a need to improve understanding of the effect of chlorine disinfection on antibiotic resistance genes (ARGs) in order to advance relevant drinking water, wastewater, and reuse treatments. However, few studies have explicitly assessed the physical effects on the DNA. Here we examined the effects of free chlorine (1–20 mg Cl₂/L) on extracellular genomic, plasmid DNA and select ARGs. Chlorination was found to decrease the fluorometric signal of extracellular genomic and plasmid DNA (ranging from 0.005 to 0.05 mg/mL) by 70%, relative to a no-chlorine control. Resulting DNA was further subject to a fragment analysis using a Bioanalyzer, indicating that chlorination resulted in fragmentation. Moreover, chlorine also effectively deactivated both chromosomal- and plasmid-borne ARGs, mecA and tetA, respectively. For concentrations >2 mg Cl₂//L × 30 min, chlorine efficiently reduced the qPCR signal when the initial concentration of ARGs was 10⁵ copies/mL or less. Notably, genomic DNA and mecA gene signals were more readily reduced by chlorine than the plasmid-borne tetA gene (by ~2 fold). Based on the results of qPCR with short (~200 bps) and long amplicons (~1200 bps), chlorination could destroy the integrity of ARGs, which likely reduces the possibility of natural transformation. Overall, our findings strongly illustrate that chlorination could be an effective method for inactivating extracellular chromosomal- and plasmid-borne DNA and ARGs.     

Impacts of advanced treatment processes on elimination of antibiotic resistance genes in a municipal wastewater treatment plant

The distributions of ARGs were monitored in a WWTP in Harbin during six months. CASS had the best removal efficacy of ARGs compared to other processes in the WWTP. UV disinfection could effectively control the HGT. AGAC significantly remove ARGs and organics due to its high absorption capacity. Combination of ozone and AGAC significantly improve removal of ARGs and organics. Antibiotic resistance genes (ARGs) pose a serious threat to public health. Wastewater treatment plants (WWTPs) are essential for controlling the release of ARGs into the environment. This study investigated ARG distribution at every step in the treatment process of a municipal WWTP located in Harbin for six consecutive months. Changes in ARG distribution involved in two advanced secondary effluent treatment processes, ozonation and granular activated carbon (GAC) adsorption, were analyzed. Biological treatment resulted in the highest ARG removal (0.76–1.94 log reduction), followed by ultraviolet (UV) disinfection (less than 0.5-log reduction). Primary treatment could not significantly remove ARGs. ARG removal efficiency increased with an increase in the ozone dose below 40 mg/L. However, amorphous GAC (AGAC) adsorption with a hydraulic retention time (HRT) of 1 h showed better removal of ARGs, total organic carbon (TOC), total nitrogen (TN), and total phosphorus (TP) than ozonation at a 60 mg/L dose. UV treatment could efficiently reduce the relative ARG abundance, despite presenting the lowest efficiency for the reduction of absolute ARG abundance compared with GAC and ozone treatments. The combination of ozone and AGAC can significantly improve the removal of ARGs, TOC, TN and TP. These results indicate that a treatment including biological processing, ozonation, and AGAC adsorption is a promising strategy for removing ARGs and refractory organic substances from sewage.     

饮用水有机类消毒副产物毒理学研究方法进展

伴随着饮用水消毒技术的改进,有机类消毒副产物(DBPs)的种类日趋多样化,其生物毒性和健康风险受到广泛关注。基于国内外对DBPs毒理学效应的研究成果,结合消毒技术的发展历程,对有机类DBPs进行了系统的分类,针对含碳消毒副产物(C-DBPs)和含氮消毒副产物(N-DBPs)中典型的DBPs的毒理学研究方法进行了详细的介绍,在此基础上分析了相关研究的不足之处和发展趋势,对今后的研究方向提出了建议。     

纳米材料对环境抗生素抗性基因污染扩散影响的研究进展

抗生素抗性基因(antibiotic resistance genes,ARGs)的环境扩散严重威胁了人类健康和生态安全。除抗生素滥用所产生的选择性压力以外,其他环境物质也能影响ARGs的传播。而纳米材料的广泛应用使其不可避免地在环境中扩散并进而影响ARGs的环境分布。因此,笔者综述了近年来纳米材料影响ARGs污染扩散的研究,并探讨了纳米材料对ARGs传播的影响机制,旨在深入理解ARGs的环境扩散行为,为ARGs环境控制及纳米材料非毒性环境效应的评估提供理论和技术支持。     

Mitigating microbiological risks of potential pathogens carrying antibiotic resistance genes and virulence factors in receiving rivers: Benefits of wastewater treatment plant upgrade

● Abundance of MAGs carrying ARG-VF pairs unchanged in rivers after WWTP upgrade. ● Upgrade of WWTPs significantly reduced diversity of pathogenic genera in rivers. ● Upgrade of WWTPs reduced most VF (ARG) types carried by potential pathogens in rivers. ● Upgrade of WWTPs narrowed the pathogenic host ranges of ARGs and VFs in rivers. Wastewater treatment plants (WWTPs) with additional tertiary ultrafiltration membranes and ozonation treatment can improve water quality in receiving rivers. However, the impacts of WWTP upgrade (WWTP-UP) on pathogens carrying antibiotic resistance genes (ARGs) and virulence factors (VFs) in rivers remain poorly understood. In this study, ARGs, VFs, and their pathogenic hosts were investigated in three rivers impacted by large-scale WWTP-UP. A five-year sampling campaign covered the periods before and after WWTP-UP. Results showed that the abundance of total metagenome-assembled genomes (MAGs) containing both ARGs and VFs in receiving rivers did not decrease substantially after WWTP-UP, but the abundance of MAGs belonging to pathogenic genera that contain both ARGs and VFs (abbreviated as PAVs) declined markedly. Genome-resolved metagenomics further revealed that WWTP-UP not only reduced most types of VFs and ARGs in PAVs, but also effectively eliminated efflux pump and nutritional VFs carried by PAVs in receiving rivers. WWTP-UP narrowed the pathogenic host ranges of ARGs and VFs and mitigated the co-occurrence of ARGs and VFs in receiving rivers. These findings underline the importance of WWTP-UP for the alleviation of pathogens containing both ARGs and VFs in receiving rivers.     

管网生物膜菌株胞外聚合物的提取方法比较

以饮用水管网生物膜样品中一株强成膜能力的菌株Pleomorphomonas oryzae作为研究对象,考察了8种方法(高速离心法、超声法、加热法、EDTA法、H2SO4法、NaOH法、SDS法、甲醛法)对菌株胞外聚合物(EPS)的提取效果,并结合三维荧光光谱(EEM)和傅立叶红外光谱(FTIR)对提取的EPS进行成分分析.结果表明,EDTA法和H2SO4法既能提高EPS的提取效率,提取量分别为64.77 mg.g-1SS和74.43 mg.g-1SS,是离心方法提取量的1.62倍和1.86倍,又不会在提取过程中对菌株细胞造成破坏,是较为理想的EPS提取方法.EEM分析进一步证实,NaOH法对菌株细胞破坏严重,造成EPS成分变化较大.FTIR分析则说明,化学提取方法相较于物理提取方法会引入杂质对组分测定造成干扰.