Mobile genetic elements facilitate the transmission of antibiotic resistance genes in multidrug-resistant Enterobacteriaceae from duck farms

Multidrug-resistant(MDR)Enterobacteriaceae critically threaten duck farming and public health.The phenotypes,genotypes,and associated mobile genetic elements(MGEs)of MDR Enterobacteriaceae isolated from 6 duck farms in Zhejiang Province,China,were investigated.A total of 215 isolates were identified as Escherichia coli(64.65%),Klebsiella pneumoniae(12.09%),Proteus mirabilis(10.23%),Salmonella(8.84%),and Enterobacter cloacae(4.19%).Meanwhile,all isolates were resistant to at least two antibiotics.Most isolates carried tet(A)(85.12%),blaTEM(78.60%)and sul1(67.44%)resistance genes.Gene co-occurrence analysis showed that the resistance genes were associated with IS26 and integrons.A conjugative IncFII plasmid pSDM004 containing all the above MGEs was detected in Proteus mirabilis isolate SDM004.This isolate was resistant to 18 antibiotics and carried the blaNDM-5 gene.MGEs,especially plasmids,are the primary antibiotic resistance gene transmission route in duck farms.These findings provide a theoretical basis for the rational use of antibiotics in farms which are substantial for evaluating public health and food safety.

Performance Parameters:Demobilization Antibiotic Resistant Bacteria(ARB)and Carrying Genes(ARG)in Wastewater Disinfection

The UV irradiation is used for removing Antibiotic Resistant Bacteria(ARB)and Antibiotic Resistance Genes(ARG)from wastewater treatment.Bacteriophages are viruses that infect within bacteria,are recognized for bacterial control.The influence of some parameters in quantification and performance influencing of pathogen demobilization could be considered in disinfection of wastewater.The comparison of Polyvalent phage(NE1)versus Coliphage(NE4)in suppressing a bacterium Escherichia coli(NDM-1:b-lactam-resistant)with UV irradiation was observed the efficacy in reduction of cells in the disinfection and parameter process.The results with the effect of UV-C irradiation on NDM-1 infected with 1%of NE4 showed a decrease of cells from 8×10^(6)to 2×10^(5)in 60 min with UV-C dose.The NDM1(E.coli)was infected with 1%of NE4(Polyvalent Phage)under magnetic stirring for 1 h,the cells count was 8×10^(6).After 1 h in UV-C e×posure,the cells number reached 3×10^(5).The NDM1 that was e×posed in 1 h of UV-C irradiation and then was infected with 1%of NE4.Cells counting were done 24 h after this procedure.These cells were e×posed in UV-C and showed a reduction in the number of cells from 1×10^(8)to 4×10^(5)after 60 min.The results indicate that bacteriophages can mitigate bacteria species,and combined the conventional water disinfection technologies that can support the microbial safety control strategies.

Trifunctional Cu-Mesh/Cu_(2)O@FeO Nanoarrays for Highly Efficient Degradation of Antibiotic, Inactivation of Antibiotic-Resistant Bacteria, and Damage of Antibiotics Resistance Genes

Trifunctional Cu-mesh/Cu_(2)O@FeO nanoarrays heterostructure is designed and fabricated by integrating CuCu_(2)O@FeO nanoarrays onto Cu-mesh(CM)via an in situ growth and phase transformation process.It is successfully applied to efficiently mitigate the antibiotic pollution,including degradation of antibiotics,inactivation of antibiotic-resistant bacteria(ARB),and damage of antibiotics resistance genes(ARGs).Under visible-light irradiation,CM/CuCu_(2)O@FeO nanoarrays exhibit a superior degradation efficiency on antibiotics(e.g.,up to 99%in 25 min for tetracycline hydrochloride,TC),due to the generated reactive oxygen species(ROS),especially the dominant·O^(2−).It can fully inactivate E.coli(HB101)with initial number of~108 CFU mL^(−1) in 10 min,which is mainly attributed to the synergistic effects of 1D nanostructure,dissolved metal ions,and generated ROS.Meanwhile,it is able to damage ARGs after 180 min of photodegradation,including tetA(vs TC)of 3.3 log 10,aphA(vs kanamycin sulfate,KAN)of 3.4 log 10,and tnpA(vs ampicillin,AMP)of 4.4 log 10,respectively.This work explores a green way for treating antibiotic pollution under visible light.

微藻生物系统去除抗生素和ARGs的研究进展

抗生素与抗生素抗性基因(ARGs)污染严重危害人类健康,微藻生物技术在处理抗生素废水过程中表现出良好的效果。文章首先总结了微藻对于不同种类抗生素的去除机理,包括生物吸附、生物累积、生物降解、光生物降解和水解挥发5种。分析了微藻在去除抗生素过程中的重要影响因素,包括pH、水力停留时间、光照、抗生素的种类与浓度和微藻的种类。并总结了微藻对ARGs的去除机制,包括移动基因元件的去除及活性氧自由基的猝灭。最后,文章指出了微藻处理抗生素废水过程中仍然存在的问题,进一步明确了今后微藻生物技术处理抗生素废水的重点研究方向。

UV-Based Advanced Oxidation Processes for Antibiotic Resistance Control: Efficiency, Influencing Factors, and Energy Consumption

Antibiotic resistant bacteria(ARB)with antibiotic resistance genes(ARGs)can reduce or eliminate the effectiveness of antibiotics and thus threaten human health.The United Nations Environment Programme considers antibiotic resistance the first of six emerging issues of concern.Advanced oxidation processes(AOPs)that combine ultraviolet(UV)irradiation and chemical oxidation(primarily chlorine,hydrogen peroxide,and persulfate)have attracted increasing interest as advanced water and wastewater treatment technologies.These integrated technologies have been reported to significantly elevate the efficiencies of ARB inactivation and ARG degradation compared with direct UV irradiation or chemical oxidation alone due to the generation of multiple reactive species.In this study,the performance and underlying mechanisms of UV/chlorine,UV/hydrogen peroxide,and UV/persulfate processes for controlling ARB and ARGs were reviewed based on recent studies.Factors affecting the process-specific efficiency in controlling ARB and ARGs were discussed,including biotic factors,oxidant dose,UV fluence,pH,and water matrix properties.In addition,the cost-effectiveness of the UV-based AOPs was evaluated using the concept of electrical energy per order.The UV/chlorine process exhibited a higher efficiency with lower energy consumption than other UV-based AOPs in the wastewater matrix,indicating its potential for ARB inactivation and ARG degradation in wastewater treatment.Further studies are required to address the trade-off between toxic byproduct formation and the energy efficiency of the UV/chlorine process in real wastewater to facilitate its optimization and application in the control of ARB and ARGs.

Fate and Behavior of Tetracycline Resistance Genes in Activated Carbon Adsorption

The accessibility of tetracycline resistance gene (tetG) into the pores of activated carbon (AC), as well as the impact of the pore size distribution (PSD) of AC on the uptake capacity of tetG, were investigated using eight types of AC (four coal-based and four wood-based). AC showed the capability to admit tetG and the average reduction of tetG for coal-based and wood-based ACs at the AC dose of 1 g·L-1 was 3.12 log and 3.65 log, respectively. The uptake kinetic analysis showed that the uptake of the gene followed the pseudo-second-order kinetics reaction, and the uptake rate constant for the coal-based and wood-based ACs was in the range of 5.97 × 10-12 - 4.64 × 10-9 and 7.02 × 10-11 - 1.59 × 10-8 copies·mg-1·min-1, respectively. The uptake capacity analysis by fitting the obtained experiment data with the Freundlich isotherm model indicated that the uptake constant (KF) values were 1.71 × 103 - 8.00 × 109 (copies·g-1)1-1/n for coal-based ACs and 7.00 × 108 - 3.00 × 1010 (copies·g-1)1-1/n for wood-based ones. In addition, the correlation analysis between KF values and pore volume as well as pore surface at different pore size regions of ACs showed that relatively higher positive correlation was found for pores of 50 - 100 Å, suggesting ACs with more pores in this size region can uptake more tetG. The findings of this study are valuable as reference for optimizing the adsorption process regarding antibiotic resistance-related concerns in drinking water treatment.

Dissemination of Resistance Integrons and Genes Coding for Blse and Cabapenemases in the Urban Drainage Network in Cote d’Ivoire

Antibiotic resistance has become a major threat to human health worldwide. Environment, particularly the water environment, has long been overlooked as a player in the antibiotic resistance cycle, although its role remains unclear. These can provide an ideal setting for the acquisition and dissemination of antibiotic resistance, as they are frequently affected by anthropogenic activities. The objective of this study was to establish a diffusion map of resistance integrons used as genetic markers of resistance associated with antibiotic resistance conferring genes (ARGs). Total DNA extracts from non-cultivable bacterial communities were used for the analyses. These communities were obtained from wastewater samples from 14 sites upstream and downstream of drainage channels or effluents in the cities of Abidjan, Bouaké, and Yamoussoukro. The results obtained correspond to the number of positives among the treated samples (n = 39). Among the genetic markers of dissemination, class 1 integrons were the most evident in 94.8% of samples in Abidjan (93.3%), Bouaké (100%) and Yamoussoukro (91.6%). Class 2 integrons and class 3 integrons were found respectively in 41% and 51% of all samples. Genes coding for β-lactamases and blaTEM was identified in almost all samples at a rate of 97.4%. A co-presence of the three genes blaTEM, blaSHV and blaCTX-M is also remarkable in the sites of the city of Yamoussoukro. Among the genes coding for carbapenemases, only blaKPC 17.94%, blaNDM 30.76% and blaOXA48 38.46% were detected in the samples.

环境中抗生素抗性基因(ARGs)形成和传播机制研究进展

从DNA损伤和错误修复角度阐述了环境中抗生素抗性基因(Antibiotics resistance genes,ARGs)的形成机制,并对DNA损伤和错误修复过程中涉及的相关核心功能基因片段进行归纳总结。也从细胞代谢角度阐述了环境中ARGs进行水平传播的三种主要机制,包括:菌毛架桥作用、信号传导和IV型细胞分泌系统。现有研究进一步加强对ARGs在环境中形成和传播机制的认识,为今后研究ARGs的削减提供理论依据。目前关于抗生素诱导ARGs形成和传播的内在机制仍相对缺乏,后续可从生物毒性和分子生物学层面进行深入研究。

The Role of Wastewater Treatment Plants in the Environmental Dissemination of Antibiotic Resistant Bacteria (ARB) and Resistance Genes (ARG)

This study was conducted to evaluate the influence of wastewater treatment processes on the prevalence of antibiotic resistance fecal coliform (FC) and antibiotic resistance genes (ARGs) of FC. In addition, the occurrence of antibiotic resistant bacteria (ARB) and antibiotic resistant genes (ARGs) in surface waters receiving wastewater was evaluated. Greater resistance against penicillin (P), colisitin (CT) and ampicillin (AMP) were observed for FC isolated from effluent disinfected by chlorine (71%), than that disinfected by UV (45%). The greatest resistance against six antibiotics was recorded for FC isolates from effluent disinfected by chlorine. The prevalence of tetB and blaSHV was lowest in isolates from chlorine-disinfected effluents. The occurrence of ARG blaSHV was highest in FC isolated from effluent disinfected by UV. A significant correlation was recorded between FC levels in surface waters and the level of bacterial resistance to ampicillin (P SHV in effluents and in surface waters. TetA and tetC were highly prevalent in surface water compared to tetB. The results of the study demonstrate the widespread prevalence of ARB and ARG in wastewater and receiving water bodies. The result indicates that the source of ARB and ARG in surface waters originate from wastewater. Released ARB and ARG may serve as the source of ARG to pathogenic bacteria in surface waters. Disinfection processes may influence the selection of antibiotic resistant patterns of bacteria.

沈阳典型粳稻种植区稻田土壤ARGs和MGEs污染分布特征分析

农田土壤中抗生素抗性基因(ARGs)与可移动遗传元件(MGEs)的污染问题,在学术界已引发广泛关注。为研究沈阳市粳稻种植区稻田土壤中ARGs与MGEs的存在和相关性,并探讨其对土壤-植物系统的潜在影响,通过采集沈阳市辽中区、新民市、沈北新区和苏家屯区的粳稻种植区域土壤样品,利用高通量qPCR技术,对285个ARGs、10个MG-Es和一个16S rRNA基因进行检测和定量分析。结果表明:在不同地区的稻田土壤中共检测到273种ARGs和MGEs,且种类和丰度存在显著差异。进一步的相关性分析发现,ARGs与MGEs之间存在一定相关性,并且这种相关性在不同地区间有所差异。多种ARGs与MGEs之间存在显著正相关(p<0.05),其中部分ARGs[tet(32)等]与MGEs(tnpA-01等)存在及极显著正相关(p<0.01),这表明在土壤中MGEs对ARGs迁移和扩散起到促进作用。4个采样区根际土中ARGs和MGEs的总相对丰度均高于非根际土壤,这进一步凸显根际环境在ARGs和MGEs分布中的重要性。此外,还发现在不同地区的稻田土壤中,抗生素失活机制在ARGs中占据主导地位。综上所述,试验结果为深入了解东北水稻种植区稻田土壤中ARGs的污染现状及其潜在影响提供了数据支持和理论基础。这些发现不仅有助于认识农田土壤中ARGs和MGEs的分布与迁移规律,还为制定针对性的土壤污染防控措施提供了科学依据。

除草剂对质粒介导ARGs接合转移的影响及机制

本研究以同时携带四环素、氨苄青霉素和卡那霉素3种抗生素抗性基因(ARGs)的可移动质粒RP4为供体质粒(保存于供体菌E.coli DH5α中),以E.coli HB101为受体菌,以丁草胺、百草敌和草铵膦作为除草剂代表,研究了不同浓度除草剂对可移动质粒介导的ARGs接合转移的影响,并从细菌细胞膜通透性、胞内ROS含量以及与接合转移相关调控基因的转录水平等角度,阐明了其作用机制.结果表明,100~800mg/L的丁草胺以及40~160mg/L的百草敌都可促进质粒RP4的接合转移.供试浓度范围内,丁草胺皆可刺激细菌胞内产生ROS,百草敌则使ROS水平下降.丁草胺和百草敌处理下,受体菌表面皆出现明显的皱缩,且随着浓度升高,出现孔洞甚至断裂现象.400~800mg/L的丁草胺和40~80mg/L的百草敌皆可通过提高细胞膜通透性,下调kor A基因的转录,同时上调trb Bp、trf Ap和tra A基因的转录,进而促进RP4的接合转移.供试浓度下,草铵膦对RP4的接合影响甚微,对细菌细胞膜通透性、胞内ROS水平以及接合相关调控基因转录水平的影响也不大.研究结果提示需要关注农田环境中除草剂的使用对ARGs传播的风险,并建议通过控制除草剂的种类及施用浓度来控制ARGs在农田生态系统中的接合转移.

辽宁省主要设施农业集散地大棚土壤ARGs与MGEs分布特征分析

为分析设施农业集散地大棚土壤中抗生素抗性基因(antibiotic resistance genes,ARGs)污染与可移动基因元件(mobile genetic elements,MGEs)的分布特征,于辽宁省6个主要设施农业集散地采集温室大棚土壤样品进行实验分析。由于长期施用畜禽粪肥,致使土壤中ARGs的相对丰度和多样性显著增加。对采样点的土壤进行ARGs及MGEs种类和丰度的检测,并对其相关性进行分析。结果表明:土壤样品中检测出50种ARGs和10种MGEs,其中tetG-01、tetG-02、tetM-01、tetM-02、tetPA、tetPB-03、tetT、oprJ、acrA-04、sul2、dfrA1、folA、blaTEM、fox5、ermX、ermB、ermF、ermT-02以及intI-1、tnpA-01、tnpA-05的检出率为100%,且抗性机制主要为细胞保护;土壤中部分ARGs的相对丰度与MGEs的相对丰度之间存在一定的正相关。

Analysis of antibiotic resistance genes and their effects on marine environment

The application of overgrowth antibiotic resistance genes in marine research was discussed.Using the China National Knowledge Infrastructure(CNKI)network database to retrieve papers on resistance genes(n=30627)and antibiotic resistance genes(n=3277),the published trends,subject headings and other aspects of bibliometrics were analyzed.Literature in this field has increased rapidly in the past 38 years.55.3%of the papers were published in the last ten years,31.2%of the papers were published in the last five years.The relevance of research topics has increased in recent years,the current research mainly focus on antibiotics,resistance gene,antibiotic resistance gene,and drug resistance.Literature analysis is helpful to understand the overall development trend of overgrowth antibiotic resistance genes in marine research.

Antibiotic resistance analysis and coping strategies of helicobacter pylori

Helicobacter pylori(H.pylori)is a Gram-negative bacterium that mainly colonizes the stomach and duodenum,and it can cause gastrointestinal diseases such as gastric inflammation,peptic ulcer and gastric cancer,and eradication of H.pylori can effectively stop the occurrence and development of gastrointestinal diseases.Antibiotics are one of the main drugs used to treat H.pylori.Due to the long-term application of antibiotics,the resistance rate of H.pylori to antibiotics increases year by year,which greatly reduces the eradication rate of H.pylori and increases the difficulty of re-treatment and the economic burden of patients.In this paper,we will review three aspects of H.pylori resistance status,resistance mechanism and treatment to provide reference for the progress of H.pylori resistance research and its treatment strategy.

非抗生素类新污染物影响质粒携带的抗生素抗性基因(ARGs)水平转移研究进展

人们通常认为抗生素的选择压力是造成抗生素抗性基因快速扩散的原因,但是越来越多的研究表明环境中非抗生素类新污染物也能够造成抗生素抗性基因快速扩散。本文对非抗生素类新污染物影响质粒携带抗性基因水平转移规律和机制研究进展进行了归纳总结。目前的研究大多集中在内分泌干扰物、药品及个人护理产品以及纳米材料影响R质粒携带抗生素抗性基因水平转移,相关机制主要关注非抗生素类新污染物对活性氧、应激反应以及细胞膜通透性的影响。持久性有机污染物影响质粒携带抗性基因水平转移规律以及非抗生素类新污染物对其他质粒携带的抗生素抗性基因水平转移规律和其他类型的机制可以作为未来的研究方向。

饮水系统中抗生素抗性基因赋存特征及健康风险评估

近几年抗生素抗性基因(ARGs)作为一种新型污染物在世界各地的水体、土壤中被频繁检出,其在环境中大量扩散和增殖十分容易导致微生物获得抗生素抗性,对人体健康产生潜在威胁。结合国内外文献报道数据,介绍了ARGs在城市饮水系统中污染现状,描述了其赋存特征,发现在国内外城市饮水系统中ARGs的数量不容小觑,数量最高可达1.38×105 copies·ml-1。其次对城市饮水系统中常用工艺对ARGs的赋存与传播影响因素进行了总结,发现城市常用水处理工艺对ARGs的灭杀效果差,甚至对其富集和繁殖往往起着促进作用;在不同的环境影响因素中,微生物群落结构是影响ARGs的主要驱动力,其次是重金属。最后提出ARGs对人体可能造成的健康威胁以及现有健康风险评估方法的局限性,并对未来ARGs的研究进行了展望。

控制医院污水中抗生素耐药细菌传播的消毒技术研究进展

全球范围内抗生素抗药菌(ARB)和抗药基因(ARGs)的扩散已成为威胁人类健康的重要问题。医院是抗生素污染的集中之地,其排放的污水是耐药菌传播的重要途径。因此,应用绿色、高效的消毒技术对于控制抗生素耐药细菌传播意义重大。本文综述了医院污水中抗生素耐药性细菌传播主要消毒技术的研究进展,这些技术不仅包括传统氧化工艺,还包括高级氧化工艺(AOPs);介绍了这些技术的应用和研究现状,分析了其去除抗药致病菌和抗药基因的性能;并对未来消毒技术的研究和应用方向提出了展望。

施用猪粪水对青脆李果实品质和产量、土壤中微生物群落与抗生素耐药基因的影响

【目的】为了解猪粪浇灌对青脆李果实品质和产量、土壤性质、土壤中微生物群落与抗生素耐药基因的影响。【方法】以5年生青脆李作试材,设置猪粪水按不同比例(100%、75%、50%、25%、0%)替代化肥田间试验,通过单果重、单果横径等外观指标,李子每667 m^(2)产量,维生素C、可溶性固形物等果实品质指标测定,分析不同施肥处理对青脆李果实品质和产量的影响;基于土壤中pH、有机质含量等理化指标测定,分析不同施肥处理对李树土壤理化性质的影响;基于Illumina Hiseq 4000测序平台对李树土壤宏基因组测序,分析猪粪水浇灌对土壤中微生物群落变化、抗生素耐药基因分布的影响。【结果】不同施肥处理青脆李果实品质和产量研究结果表明,“75%猪粪+25%化肥”“50%猪粪+50%化肥”处理青脆李果实外观品质均较优,“50%猪粪+50%化肥”处理青脆李每667 m^(2)产量最高。不同施肥处理对李树土壤理化性质的研究结果显示,各处理间有机质、碱解氮等理化指标均不存在显著性差异(P>0.05)。宏基因组测序分析结果显示,“100%猪粪”组中的优势菌群为放线菌,而其余4组的优势菌群均为假单胞菌门;共鉴定39个抗生素本体(ARO),随着猪粪水用量的减少,土壤样品中检出ARO种类、ARO相对丰度和均呈依次递减趋势。【结论】本研究条件下,猪粪水部分替代化肥为青脆李树施肥是可行的,且以“50%猪粪+50%化肥”进行施肥最佳。

商品灭菌乳中常见抗生素抗性基因的筛查与定性分析

探究了市售常见商品灭菌乳中四环素类、氨基糖苷类、β-内酰胺类等抗生素抗性基因(Antibiotic resistance genes,ARGs)的分布与带染特征。以12种市售常见商品灭菌乳为研究对象,用CTAB法提取预处理后样品中的总DNA;超微量分光光度计测量DNA纯度与浓度;PCR扩增ARGs;PCR产物测序后提交GenBank进行序列比对,分析ARGs的种类;使用SPSS软件对ARGs检出率、样品品牌、包装类型、产品类型等进行相关性分析。结果显示,灭菌乳中可同时检出多种ARGs,检出率较高的基因分别为(str B(93.75%)、aad B (92.71%)、sul1(85.42%)、bla_(TEM)82.29%)和tet C(58.33%),未在35种ARGs中检出bla_(VEB)、bla_(PER)、bla_(DHA)、bla_(CMY2)和bla_(GIM)。商品灭菌乳的品牌、包装、产地和保质期与aad A、aad B、str A、str B等抗性基因检出率显著相关。市售商品灭菌乳中存在多种ARGs,且不同种类样品间存在差异,值得从食品安全角度深入研究。

畜禽粪肥中抗生素残留对土壤微生物及抗性基因的影响分析研究进展

简述了施用集约化养殖的畜禽粪肥对农田土壤造成不利影响的抗生素残留水平和种类,并结合抗生素检测方法的发展对国内外农田土壤抗生素残留现状进行分析。探究了抗生素对土壤微生物菌群的影响,揭示了抗生素抗性基因(ARGs)对农田土壤的潜在风险。最后展望了未来应深入了解土壤微生物和ARGs之间的作用机理,从而为管控抗生素的负面环境影响提供科技支撑。