水产养殖环境中抗生素抗性基因（ARGs）的研究及进展(Research Advancement of Antibiotics Resistance Genes（ARGs）in Aquaculture Environment)

抗生素在现阶段的水产养殖中具有不可替代的作用,但长期滥用抗生素会诱导水生动物体内产生携带抗性基因(ARGs)的细菌株,尤其是多重耐药性菌株的产生将使得各种疾病的治疗更加棘手.抗性基因一旦被排泄到水产环境中不仅会对养殖区域和周围的环境造成潜在的基因污染,而且还会通过各种可移动遗传元件如质粒、转座子、整合子等的水平迁移作用进入其他致病菌和环境细菌中,对人类的健康安全构成潜在的威胁.论文对抗生素抗性基因在水产养殖业中的来源、污染现状、潜在的传播途径和相关的检测方法进行了综述,指出了开展水产养殖业中抗生素抗性基因污染研究的必要性,建议政府和有关部门尽快进行水产养殖业抗生素抗性基因的污染机理与控制对策研究.     

抗生素和纳米银对大肠杆菌的联合毒性和抗性突变诱导效应

抗生素和纳米银因在各自领域的大量应用而不可避免地进入环境中，对生态环境尤其是微生物群体构成联合暴露的风险，但是二者对细菌生长和耐药性的联合作用，目前受到的关注较少.本研究测定了两种抗生素（四环素和氯霉素）以及纳米银（10—15 nm）对大肠杆菌的联合毒性及对细菌抗性突变的诱导作用，并对相关机制进行了初步探讨.结果表明，抗生素和纳米银对细菌的联合毒性呈现协同效应，但是二者联合作用下细菌对利福平的抗性突变频率显著降低. DNA测序和分子对接结果表明，编码利福平靶标蛋白的rpoB基因发生点突变，导致突变后的靶蛋白与利福平的结合能降低.同时，qPCR结果表明抗生素和纳米银联合作用时，细菌胞内rpoS、dinB和mutS等基因表达显著上调，可能是抗性突变频率在联合作用下降低的主要原因.本研究揭示了抗生素和纳米银联合作用对细菌抗性突变的影响，有利于全面认识二者联合暴露的环境和健康风险.     

抗生素抗性基因在环境中的来源、传播扩散及生态风险

近年来,由于抗生素的滥用首先诱导动物体内产生抗生素抗性基因(antib iotic resistance genes,ARGs),从而加速了抗性基因在环境中细菌间的传播扩散.目前,抗生素抗性基因作为一类新型环境污染物,在不同环境介质中的传播、扩散可能比抗生素本身的环境危害更大.本文针对抗生素抗性基因在地表水、地下水、医疗废水、城市污水处理厂、养殖场、土壤、沉积物以及大气环境中的来源、分布、传播情况以及国内外最新研究动态进行综述.分析了抗生素抗性基因在环境中的潜在传播途径及其可能影响因素,并指出光照,厌氧,高温处理可以有效遏制抗生素抗性基因在环境中的传播和扩散.揭示了抗生素抗性基因可能造成的生态风险,针对我国对该类污染物的研究现状,提出了今后的研究重点.     

抗生素抗性基因在典型行业和市政污水中的污染特征及消减研究进展

抗生素在养殖、制药及医疗卫生行业中的广泛使用导致环境中的抗性细菌(Antibiotic resistance bacteria,ARB)及抗性基因(Antibiotic resistance genes,ARGs)浓度日益增加,甚至已经威胁到人体健康.本文在总结大量文献的基础上,阐述了ARB和ARGs在抗生素典型行业(包括养殖业、制药业和医疗卫生业)和市政污水中的污染特征,重点介绍了污水处理系统(Wastewater treatment plants,WWTPs)不同处理工艺对抗性细菌和抗性基因消长的影响及机制,最后针对污水处理系统中抗性基因的消减提出了新的见解,并对今后研究方向进行了展望.     

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。     

海洋环境中抗生素存在与环境行为研究进展

由于抗生素的频繁使用和排放，抗生素不断排放进入水环境，表现为“持续存在”的状态，对生态环境系统造成长期、持续的环境风险，由此导致的环境污染和细菌耐药性是我国及全球都亟待解决的重大环境问题.抗生素持久性的选择压力会产生抗生素抗性基因（antibiotic resistant genes,ARGs），其在环境中的持久性残留，在菌群间的迁移、传播和扩增，比抗生素残留本身对生态环境的危害更大.随着海洋资源与环境的持续开发与利用，海洋正逐步成为抗生素和ARGs的重要储存库，海洋环境中的抗生素和ARGs对海洋生态环境和公众健康的威胁同样不容忽视.本文对海洋环境中抗生素的来源、赋存、迁移转化和生态风险等研究进行分析梳理，对海洋环境中ARGs的环境行为进行综述，以期为海洋环境中抗生素和ARGs环境行为和风险评估研究提供参考，最后对相关研究的发展趋势进行了展望.     

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。     

中国边缘海沉积物中抗生素抗性基因和致病菌的组成特征

抗生素抗性基因作为一种新型“污染物”已经受到广泛关注.研究边缘海中抗生素抗性基因和人类致病菌的组成特征,有助于深入认识海洋环境中抗生素抗性基因的起源和海洋致病菌的潜在健康风险.使用基于高通量测序的宏基因组学分析方法在中国边缘海沉积物中发现了多种抗生素抗性基因,主要以多重耐药基因为主,其中南海沉积物中抗生素抗性基因的总丰度约是黄渤海的2倍.与中国边缘海相比,珠江口沉积物中抗生素抗性基因丰度更高,与人类常用抗生素（如磺胺类、氨基糖苷类、β-内酰胺类、四环素类等）的耐药性更为相关.此外,中国边缘海沉积物中几乎没有发现质粒携带的抗性基因,而珠江口有约20%的抗性基因由质粒携带.中国边缘海沉积物中主要人类致病菌种包括肺炎链球菌（24.4%）、肺炎克雷伯菌（19.9%）、无乳链球菌（9.2%）、铜绿假单胞菌（6.9%）等.珠江口沉积物中致病菌群落组成与中国边缘海显著不同,副溶血性弧菌和空肠弯曲杆菌的丰度相对更高.上述研究表明,中国边缘海沉积物中存在高多样性的抗生素抗性基因和人类致病菌,人类活动干扰可导致其污染水平提高.     

生物炭添加对土壤中抗生素和抗性基因的环境行为影响研究进展

近年来,土壤抗生素和抗性基因污染已成为我国新兴的环境问题,生物炭作为土壤改良剂施用到土壤后会影响抗生素和抗性基因的环境行为.本文从我国土壤中抗生素和抗性基因污染现状和潜在风险出发,概述了生物炭添加土壤对抗生素的吸附、解吸及老化的影响,分析了生物炭特性、土壤类型、抗生素种类,和温度、pH值、共存物质等吸附条件对生物炭添加土壤吸附抗生素的影响,阐述了生物炭添加对土壤中抗生素和抗性基因迁移、消散、生物有效性,以及酶和微生物的影响,并对生物炭控制土壤中抗生素和抗性基因的研究前景进行了展望,拟为土壤中生物炭调控技术的发展提供参考.     

抗生素在地下水系统中的环境行为及生态效应研究进展

抗生素在环境中的残留已引起广泛关注。随着对地下水污染的报道日益增多,抗生素对地下水系统的潜在影响不容忽视。本文系统地阐述了地下水中抗生素的来源、污染水平及迁移转化规律,总结了抗生素对地下水微生物群落的影响以及抗生素诱导产生的抗性基因的潜在污染趋势。因地下水赋存隐蔽,一旦污染难以及时察觉,抗生素进入地下水系统后易长期残留。目前,针对抗生素及抗性基因在地下水系统中的环境行为及生态效应研究还十分有限,本文据此指出了当前形势下开展相关研究的必要性,并对今后的研究方向进行了展望。     

Association between heavy metals and antibiotic-resistant human pathogens in environmental reservoirs: A review

Heavy metals can act as co-selecting agents and promote antibiotic resistance. Most frequent resistances to heavy metals are observed for zinc and cadmium. P. aeruginosa and E. coli are commonly resistant to heavy metals and antibiotics. Heavy metals proliferate antibiotic resistance through co- and cross-resistance. Heavy metal and antibiotic resistances are common near anthropogenic activities. Antibiotic resistance in human pathogens can proliferate under selective pressures. Heavy metals in environmental reservoirs may contribute to selecting antibiotic-resistant strains. To determine the associations between heavy metals and antibiotic resistance, a literature review was conducted to systematically collect and categorize evidence for co-occurrence of resistance to heavy metals and antibiotics within human pathogenic bacteria in water, wastewater, and soil. In total, 42 publications adhered to inclusion criteria. Across the reservoirs, zinc and cadmium were the most commonly observed heavy metals associated with resistance to antibiotics. Pseudomonas aeruginosa and Escherichia coli were the most commonly studied bacteria with reported co-occurrence of resistance to several heavy metals and antibiotic classes. As co-selecting agents, prevalence of heavy metals in the environment can proliferate resistance to heavy metals and antibiotics through co-resistance and cross-resistance mechanisms. In comparing different reservoirs, soils and sediments harbor higher heavy metal and antibiotic resistances compared to water environments. Additionally, abiotic factors such as pH can affect the solubility and hence, the availability of heavy metals to bacterial pathogens. Overall, our review demonstrates heavy metals act as co-selecting agents in the proliferation of antibiotic resistance in human pathogens in multiple environmental reservoirs. More studies that include statistical data are needed to further describe the exposure-response relationships between heavy metals and antibiotic resistance in different environmental media. Moreover, integration of culture-based and molecular-based methods in future studies are recommended to better inform our understanding of bacterial co- and cross-resistance mechanisms to heavy metals and antibiotics.     

抗生素最小抑制浓度法(MICs)评估环境大肠杆菌抗生素抗性

2015年世界卫生组织将抗生素的滥用列为21世纪最大的挑战之一,全球范围内抗生素抗药性的散播已严重威胁人类的健康。如何检测环境中的细菌抗药性,并有效评估抗药性感染的风险,是环境微生物研究的一项重要课题。本文通过改进的培养基微量稀释(broth micro-dilution)法,确定了2个地区(中国成都和美国夏威夷)不同环境来源(天然水系和市政污水)的大肠杆菌的抗生素最小抑制浓度(minimum inhibitory concentrations,MICs)。计算所得的MIC分位数(MIC50和MIC90)、菌体抗性百分比及多抗药性指数(multiple antibiotic resistance indexes,MARIs)显示两地不同环境区划的抗生素抗性存在明显的差异。天然水系(成都锦江)中的抗生素抗性是随时间可变的,与当地的降雨事件相关。环境菌株的抗药性模式通过聚类和非度量多维测度(non-metric multidimensional scaling,NMDS)进行分析。广谱β-内酰胺酶基因筛查显示出抗性基因与抗性表型之间的正相关性。结合现有的两地抗生素的使用数据讨论了两地环境抗生素抗性与当地人类活动及抗生素的使用实践之间的紧密联系。采集环境菌株抗生素MIC数据的实验及数据分析方法实现了环境抗药性的跨时空对比,为规范抗生素的区域性使用提供了指导作用。     

土壤环境中四环素类抗生素残留及抗性基因污染的研究进展(Tetracycline Residues and Tetracycline Resistance Gene Pollution in Soil: A Review)

近年来,致病菌耐药性的增加和扩散已成为全世界关注的热点问题,而人类医疗和畜禽养殖业抗生素的滥用正不断加剧这一问题.众多研究表明土壤环境作为一个巨大的抗性基因储存库,在抗性微生物和抗性基因的传播中起重要的作用,具有潜在的生态与健康风险.在总结国内外最新研究基础上,对土壤环境中典型抗生素-四环素类抗生素的主要污染源以及其在土壤中的基本环境行为等进行了分析,并探讨了土壤中四环素类抗性基因的来源、迁移和扩散及分子检测手段等问题.我国作为抗生素的生产和消费大国,抗生素污染问题较其他国家更为严重,而国内相关研究才刚刚起步,迫切需要开展有关环境中抗生素和抗生素抗性基因污染的系统研究.     

Influence of lignin, pentachlorophenol and heavy metal on antibiotic resistance of pathogenic bacteria isolated from pulp paper mill effluent contaminated river water

Pulp paper mill pollutants are the major source of aquatic contamination having metals, lignin and chlorophenols. Study was conducted to see the effect of these contaminants on antibiotic resistance pattern of isolated bacteria. Pulp paper effluents were evaluated for its physico-chemical properties i.e, BOD 72143 +/- 164.81 to 22.32 +/- 2.48, COD 213136 +/- 583.59 to 60.40 +/- 6.34, total phenol 386 +/- 71.24 to 0.43 +/- 0.0, lignin 26312 +/- 258.59 to 73.67 +/- 31.81and microbial quality i.e. K. pneumonae, S. typhi, S. faecalis, P. aeruginosa, E. coli, Ent. faecalis, A. hydrophila, B. subtilis, S. aureus, Y enterolitica and V vulrificus. Antibiotic sensitivity (10-30 microg), heavy metal resistance (100-1000 microg ml(-1)), lignin (1000-10,000 ppm) and pentachlorophenol (100-1000 ppm) tolerance of bacterial strains were assessed by seven classes of antibiotics. Eleven bacterial isolates were found multidrug resistant towards antibiotics, heavy metal, lignin and PCP. Out of 11 isolates, 90.9% were found resistant against eleven antibiotics which acquired 100% resistant in presence of heavy metal, lignin and chlorophenols. Results also revealed that concentration of lignin (50-350 ppm) and PCP (5-30 ppm) induced maximum growth (273-8050 cfu ml(-1)) of pathogenic bacteria in river water.     

Antibiotic resistance genes in manure-amended paddy soils across eastern China: Occurrence and influencing factors

• Manure fertilization resulted in antibiotic residues and increased metal contents. • The tet and sul genes were significantly enhanced with manure fertilization. • Soil physicochemical properties contributed to 12% of the variations in ARGs. • Soil metals and antibiotics co-select for ARGs. Pig manure, rich in antibiotics and metals, is widely applied in paddy fields as a soil conditioner, triggering the proliferation of antibiotic resistance genes (ARGs) in soil. However, comprehensive studies on the effects of manure fertilization on the abundance of ARGs and their influencing factors are still insufficient. Here, pig manure and manure-amended and inorganic-amended soils were collected from 11 rice-cropping regions in eastern China, and the accumulation of antibiotics, metals, and ARGs was assessed simultaneously. The results showed that manure fertilization led to antibiotic residues and increased the metal content (i.e., Zn, Cu, Ni, and Cr). Tetracycline and sulfonamide resistance genes (tetM, tetO, sul1, and sul2) were also significantly enhanced with manure fertilization. According to variance partitioning analysis, the most important factors that individually influenced ARGs were soil physicochemical properties, accounting for 12% of the variation. Significant correlations between soil nutrients and ARGs indicated that manure application enhanced the growth of resistant microorganisms by supplying more nutrients. Metals and antibiotics contributed 9% and 5% to the variations in ARGs, respectively. Their co-occurrence also increased the enrichment of ARGs, as their interactions accounted for 2% of the variation in ARGs. Interestingly, Cu was significantly related to most ARGs in the soil (r = 0.26–0.52, p<0.05). Sulfapyridine was significantly related to sul2, and tetracycline resistance genes were positively related to doxycycline. This study highlighted the risks of antibiotic and ARG accumulation with manure fertilization and shed light on the essential influencing factors of ARGs in paddy soils.     

生态毒理学教学专栏寄语

抗生素的长期滥用,使得环境细菌耐药性不断增强,加速了耐药基因(antibiotics resistance genes,ARGs)在环境中的传播扩散,并给饮用水带来健康风险。在介绍饮用水中抗生素和耐药基因污染现状的基础上,阐述了耐药基因在饮用水中的来源和归趋,对环境中抗生素和耐药基因的人体健康风险做了初步探讨,针对目前饮用水的污染现状,对今后有关饮用水中细菌耐药的研究进行了展望。     

畜禽养殖过程抗生素使用与耐药病原菌及其抗性基因赋存的研究进展

兽用抗生素在提高畜禽生产性能、防治疾病方面发挥着重要作用,目前全球超过一半以上抗生素用于畜禽养殖,畜禽养殖源耐药病原菌、抗性基因及其传播风险愈益得到人们的重视。我国是畜禽养殖和抗生素使用大国,但兽用抗生素使用、病原菌耐药水平及其抗性基因类型等数据却较为缺乏,不利于今后畜禽养殖源耐药病原菌及其传播风险的控制。因此,本文通过文献调研,对我国和主要发达国家的兽用抗生素使用情况、畜禽养殖源耐药病原菌及其携带的抗性基因、基因移动元件以及向环境传播的途径进行分析、总结,以期为规范合理用药、降低耐药病原菌及其抗性基因传播风险,建立从畜禽养殖场至公共环境全过程的抗性污染控制链条提供借鉴。     

Analysis of antibiotic resistance of Escherichia coli isolated from the Yitong River in North-east China

The concentrations of four types of antibiotics in the Yitong River were detected. The concentration of total coliforms in summer was higher than that in spring. There was a seasonal difference in antibiotic resistance of E. coli. The E. coli in the Yitong River was found to have multiple antibiotic resistance. The Yitong River is one of the largest secondary tributaries of the Songhua River. The area where the Yitong River flows is densely populated and contains the livestock and poultry breeding areas of north-east China. These areas introduce a high risk of antibiotic contamination. In this study, the concentrations of four types of typical antibiotics including quinolones, tetracyclines, sulfonamides, and trimethoprim were determined by solid phase extraction-high performance liquid chromatography. The antibiotic resistance of Escherichia coli caused by antibiotic pollution was investigated. The concentration of total coliforms in the Yitong River was detected by the plate counting method. The antibiotic resistance of E. coli to quinolones, tetracyclines, sulfonamides, and trimethoprim was analyzed by the Kirby-Bauer method. The results showed that the concentration of total coliforms in the summer was higher than that in the spring. There was a seasonal difference in the resistance rate of E. coli to antibiotics except trimethoprim. The antibiotic resistance to fluoroquinolones was relatively low. The resistance rate to tetracyclines was higher during the summer. Moreover, resistance to several antibiotics was observed in all sections. This study provides basic data for research on pollution characteristics and prevention of antibiotic exposure in rivers.     

Multiple antibiotic resistance patterns of rhizospheric bacteria isolated from Phragmites australis growing in constructed wetland for distillery effluent treatment

Susceptibility patterns of 12 different antibiotics were investigated against rhizospheric bacteria isolated from Phragmites australis from three different zones i.e. upper (0-5 cm), middle (5-10 cm), lower (10-15 cm) in constructed wetland system with and without distillery effluent. The major pollutants of distillery effluent were phenols, sulphide, heavy metals, and higher levels of biological oxygen demand (BOD), chemical oxygen demand (COD) etc. The antibiotic resistance properties of bacteria were correlated with the heavy metal tolerance (one of distillery pollutant). Twenty-two species from contaminated and seventeen species from non-contaminated site were tested by agar disc-diffusion method. The results revealed that more than 63% of total isolates were resistance towards one or more antibiotics tested from all the three different zones of contaminated sites. The multiple-drug resistance property was shown by total 8 isolates from effluent contaminated region out of which 3 isolates were from upper zone, 3 isolates from middle zone and 2 isolates were from lower zone. Results indicated that isolates from contaminated rhizosphere were found more resistant to antibiotics than isolates from non-contaminated rhizosphere. Further this study produces evidence suggesting that tolerance to antibiotics was acquired by isolates for the adaptation and detoxification of all the pollutants present in the effluent at contaminated site. This consequently facilitated the phytoremediation of effluent, which emerges the tolerance and increases resistance to antibiotics.     

气溶胶中抗生素抗性基因研究进展:以养殖场和医院为例

抗生素在医药和养殖业的大量使用导致耐药菌的出现,加速了抗生素抗性基因(antibiotic resistance genes,ARGs)在不同环境介质中的传播扩散,抗生素耐药性已成为目前全球卫生、食品安全和发展的最大威胁之一。气溶胶作为ARGs的潜在储存库缺乏系统的研究数据,而通过空气传播具有较高抗生素抗性水平的细菌可能是引起重要疾病的主要传播途径。本文针对养殖场和医院2个抗生素大量使用的典型场所,对气溶胶中ARGs的污染现状、样品采集与检测技术进行综述,并探讨了这一环境污染的潜在风险,表明开展气溶胶中ARGs研究的必要性,并为以后需开展的工作提出几点建议。