污水河道污泥中大肠杆菌抗生素抗性与金属耐受性的耦合分析

城市污水河道受人为活动的影响,可能作为抗性细菌与抗性基因的储库,给人类生产生活带来巨大的威胁.本研究以中国寒区城市污水河道污泥为取样点,研究大肠杆菌（Escherichia coli）的抗生素与金属的耐药现状,以评估中国寒区城市水环境受抗性细菌污染的情况与传播抗性的风险.利用选择性培养基从河道淤泥中分离出455株肠杆菌,并从中筛选出110株E. coli.抗生素表型结果显示：69.1%的E. coli分离株对至少1种抗生素具有抗性,其中,抗性最高的是氧四环素（42.7%）;所有的E. coli对亚胺培南和呋喃妥因没有抗性;分离株E. coli DL27对检测的17种抗生素中的13种具有抗性,分离株E. coli DL18、分离株E. coli DL46对12种抗生素具有抗性.金属抗性表型检测结果显示：40.9%的E. coli对银（Ag）具有耐受性,3.6%的分离株对汞（Hg）、锌（Zn）具有耐受性,所有菌株对铜（Cu）无耐受性;且发现分离株E. coli DL35对9种抗生素和2种金属具有抗性.对E. coli群体15种抗生素抗性基因（Antibiotic Resistance Genes,ARGs）、4种金属耐受性基因（Metal Resistance Genes,MRGs）、1种整合酶基因的检测结果显示,ARGs中tetA（56.4%）、blaTEM（26.4%）、sul2（24.5%）、sul1（22.7%）基因的检出率最高,MRGs的检出率分别为20.0%（silB）、13.6%（silE）、8.2%（merC）、16.4%（pcoE）,整合酶基因intI1的检出率为2.7%.研究表明,在河道污泥中不仅存在多重抗生素抗性E. coli,而且存在同时具有抗生素抗性和金属耐受性的E. coli.河道污泥是抗性细菌和抗性基因的储库,对人类生产生活构成了潜在的巨大威胁.     

水产细菌耐药性的最新研究概况

细菌耐药性是指细菌多次与药物接触以后,对药物的敏感性减小甚至消失,致使药物对耐药菌的疗效降低甚至无效。细菌耐药性的形成,是细菌进化发展的结果,是细菌适应性的表现,但这种适应给养殖业甚至整个人类社会构成了威胁。本文就国内外水产病原菌对不同药物的耐药性新近研究概况,包括其耐药机制、耐药基因、耐药质粒、研究方法等进行了综述,以期为科学合理的用药提供参考依据。     

在超滤草浆黑液过程中膜阻力的分布

采用截留分子量30000,20000,10000和6000的超滤膜,对草浆黑液进行了超滤过程中膜阻力分布的研究.通过测定不同条件下膜样品的通量,对聚乙二醇标准物的截留率,指出在超滤黑液过程中各部分阻力的比例和超滤膜的分离特性的变化.各种膜样品的热重分析结果表明,膜污染是由微量有机物引起的污染.     

山东地区海水养殖区常见抗生素耐药菌及耐药基因分布特征

对山东地区海水养殖海域常见抗生素耐药菌及耐药基因分布进行调查,结果表明所调查5个海水养殖区水样中四环素类、磺胺类、-内酰胺类耐药菌比例显著(p0.05)高于氯霉素类和喹诺酮类耐药菌;此外用RT-PCR方法对共计15种耐药基因在水样中的丰度测定表明磺胺类(sul、dfra/16S rRNA=10-6~10-2)、喹诺酮类(qnr/16S rRNA=10-6~10-2)以及四环素类耐药基因(tet/16S rRNA=10-7~10-2)在各水样中丰度差异不显著,而氯霉素耐药基因(cata、cmle/16S rRNA=10-8~10-2)在不同水样中丰度差异显著,且cata1和cmle1丰度与可培养氯霉素耐药菌比例存在相关线性关系。实验数据说明山东海水养殖区水样中存在一定的抗生素耐药菌和耐药基因污染。     

Film mulching reduces antibiotic resistance genes in the phyllosphere of lettuce

Phyllosphere is an important reservoir of antibiotic resistance genes(ARGs), but the transfer mechanism of ARGs from soil and air to phyllosphere remains unclear. This study demonstrated that soil-air-phyllosphere was the dominant ARG transfer pathway, and blocking it by film mulching can reduce typical phyllosphere ARGs in lettuce by 80.7%-98.7%(89.5% on average). To further eliminate phyllosphere ARGs in lettuce grown with film mulching, the internal soil-endosphere-phyllosphere transfer pathw...     

On wear resistance of tool steel

Maintaining a reasonably low cutting tool wear when producing forming tools is a general challenge in the development of new forming tool materials. The tool life of a hot forming tool steel (H13) has been significantly improved by reducing its Si-content from 1.0 to 0.06 wt.%. However, this modified H13 (MH13) also displays a reduced cutting tool life due to higher cutting forces and a stronger tendency to form built up layers (BUE) on the cutting edge. This paper explains why.Gleeble tests of MH13 revealed a significantly higher flow stress in the 820–900 °C temperature interval in MH13 compared to H13. Thermo-Calc simulations showed that when reducing the Si-content from 1.0 to 0.06 wt.% the initial temperature for ferrite-to-austenite transformation (A1) was reduced from 900 °C to 820 °C. Knowing that austenite has totally different mechanical and thermal properties than ferrite, the difference in A1 between the two steels explains the higher cutting forces and higher tendency for BUE-formation. The conclusion is that the difference in machinability between H13 and MH13 is primarily related to their difference in A1.An attempt was also made to find a new tool material composition that can combine the wear resistance of MH13 and the good machinability of H13. Thermo-Calc simulations were performed with slightly modified alloying content without changing its properties as a good forming tool material, with the aim to increase A1. For instance, reducing the Mn content from 0.5 to 0.05 wt.% proved to increase A1 from 820 to 850 °C.     

一株茎点霉菌的抗镉机制

从矿区土壤筛选到一株对镉具有较强抗性和富集能力的丝状真菌,编号为F2,根据其形态主要是分生孢子器的特征鉴定为茎点霉菌.对不同镉浓度液体培养下菌体的生长及培养液的pH值变化作了分析,利用扫描电镜、透射电镜与能谱分析相结合的方法研究镉在F2胞内外的沉积作用及分布情况,从而分析该菌对镉的抗性和富集机制.结果表明,100mg/L的镉浓度下,在细胞壁及周围区域存在大量颗粒状镉沉积物,而在菌体表面也有沉淀物附着.因此胞内外沉积作用可能是该菌对高浓度镉的抗性和富集作用的重要途径,而且因为该菌生长过程中尤其是高镉浓度下pH值的大幅度上升,沉积过程可能与该菌在生长过程中产生碱性物质有关.     

Occurrence and distribution of antibiotic resistance genes in the sediments of the East China Sea bays

The coastal area of the East China Sea has experienced rapid urbanization and industrialization in China since 1980 s, resulting in severe pollution of its environments.Antibiotic resistance genes(ARGs) are regarded as a kind of emerging pollutant with potential high risk. The sediment samples were collected from Hangzhou Bay(HB),Xiangshan Bay(XB), and Taizhou Bay(TB) to investigate the spatial occurrence and distribution of 27 ARGs and class I integron–integrase gene(intI1) in the coastal area of the East China Sea. The PCR results showed the frequent presence of 11 ARGs and intI1 in the sediments of the three bays. The qPCR results further showed that sulfonamide resistance was the most prevalent ARG type and antibiotic target replacement and protection were the most important resistance mechanisms in the sediments. Regarding the subtype of ARGs, sulI, tetW, and dfrA13 were the most abundant ARGs, in which sulI was higher in TB(based on both the absolute and relative abundances) and dfrA13 was higher in HB(based on the relative abundances). The network analysis revealed that intI1 had significant correlations with tetC, sulI, sulII, and blaPSE-1. Oil was the key connected factor, which had positive connections with sulI, sulII, and blaPSE-1. In addition, the joint effect of heavy metals and nutrients organic pollutants might be crucial for the fate of ARGs in the coastal sediments.     

微塑料影响抗性基因的传播与水平基因转移

微塑料作为一种新兴污染物,在多种环境介质（大气、土壤、水体、沉积物）中均有广泛检出.微塑料能够通过表面形成的生物膜结构携带抗性基因,对人体健康造成的潜在风险已经引起了国内外学者的广泛关注.但是,微塑料在抗性基因的长距离传播以及水平转移方面所发挥的功能,以及微塑料传播耐药性所导致的生态与环境效应还尚不明晰.本文重点对不同环境中的微塑料与抗性基因的污染现状进行了综述讨论,对微塑料传播抗性基因的主要方式与过程进行了对比分析,最后概述了微塑料参与调控抗性基因水平转移的机制,并对未来相关研究的重点方向进行了展望,以促进微塑料污染问题的科学解决.     

From chemical mixtures to antibiotic resistance

In real environment, it is unlikely that contaminants exist singly; environmental contamination with chemical mixtures is a norm. However, the impacts of chemical mixtures on environmental quality and ecosystem health have been overlooked in the past. Among the complex interactions between different contaminants, their relationship with the rise of antibiotic resistance (AR) is an emerging environmental concern. In this paper, we review recent progresses on how chemicals or chemical mixtures promote AR. We propose that, through co-selection, agents causing stress to bacteria may induce AR. The mechanisms for chemical mixtures to promote AR are also discussed. We also propose that, mechanistic understanding of co-selection of chemical mixtures for AR should be a future research priority in environmental health research.     

Characterizing the antibiotic resistance genes in a river catchment: Influence of anthropogenic activities

Previous studies on environmental antibiotics resistance genes(ARGs) have focused on the pollution sources such as wastewater treatment plants, aquaculture and livestock farms,etc. Few of them had addressed this issue in a regional scale such as river catchment. Hence,the occurrence and abundances of 23 ARGs were investigated in surface water samples collected from 38 sites which located from the river source to estuary of the Beijiang River.Among them, 11 ARGs were frequently detected in this region and 5 ARGs(sul I, sul II, tet B,tet C, and tet W) were selected for their distribution pattern analysis. The abundances of the selected ARGs were higher in the upstream(8.70 × 10~6 copies/ng DNA) and downstream areas(3.17 × 10~6 copies/ng DNA) than those in the midstream areas(1.23 × 10~6 copies/ng DNA), which was positively correlated to the population density and number of pollution sources. Pollution sources of ARGs along the Beijiang River not only had a great impact on the abundances and diversity, but also on the distribution of specific ARGs in the water samples. Both sul I and sul II were likely originated from aquaculture farms and animal farms,tet W gene was possibly associated with the mining/metal melting industry and the electric waste disposal and tet C gene was commonly found in the area with multiple pollution sources.However, the abundance of tet B was not particularly related to anthropogenic impacts. These findings highlight the influence of pollution sources and density of population on the distribution and dissemination of ARGs at a regional scale.     

厌氧和好氧处理过程中四环素抗药基因的丰度

为了了解抗生素生产废水不同处理过程(厌氧生物处理和好氧生物处理)中抗药基因的行为,本文以两种四环素(土霉素、金霉素)生产废水处理系统为调查对象,采用PCR和实时定量PCR(qPCR)方法考察厌氧和好氧处理过程中常见的6种四环素抗药基因(tet(A), tet(C), tet(G), tet(Q), tet(W), tet(X))及2种转移因子(I型整合子intI1, 异常插入序列ISCR3)的丰度特征.结果表明,tet(C)在所有样品中均未检出,其它基因在所有样品中检出.四环素生产废水处理系统中,厌氧污泥中tet(A)、tet(G)、tet(X)的相对丰度(与16S rRNA基因的比值)范围为(1.25±0.16)×10^-4~(4.52±0.002)×10^-2,显著低于好氧污泥[(9.88±0.67)×10^-5~(2.70±0.29)×10^-1],而tet(Q)、tet(W)在厌氧污泥中的相对丰度为(1.66±0.03)×10^-2~(7.48±1.22)×10^-2,比好氧污泥中[(1.94±0.12)×10^-3~(2.85±0.16)×10^-2]高1个数量级;转移因子intI1和ISCR3在厌氧污泥中相对丰度范围为(1.48±0.01)×10^-3~(2.61±0.31)×10^-2,显著低于好氧污泥[(1.18±0.15)×10^-1~(8.99±0.75)×10^-1],表明厌氧处理过程中由这两种转移因子介导的水平转移潜力较小.研究表明,好氧处理促进了tet(A)、tet(G)、tet(X)的传播,但对tet(Q)和tet(W)有控制效果,而厌氧处理过程与之相反.抗药基因的分布与水平转移因子、抗药机制、群落结构有关.     

汾河水体抗生素抗性基因赋存特征

抗生素抗性基因(Antibiotic resistance genes,ARGs)被认为是一种新型污染物,而水环境系统是ARGs释放和传播的重要媒介.为探究汾河水体环境中ARGs的赋存特征及分布特征,选取汾河水体从源头到入黄口的12个位点,对环境因子、ARGs的丰度及多样性,以及二者之间的关系进行研究.结果表明,汾河水...     

基于病毒宏基因组学鉴定水环境中噬菌体携带的新型耐药基因及其耐药特点分析

水环境作为耐药基因的存储库，为耐药基因的水平转移和新型耐药基因的产生提供重要场所.但到目前为止，对水环境中病毒携带耐药基因的特征及其功能还知之甚少.因此以南四湖和东平湖为研究对象，利用病毒宏基因组学结合体外实验的方法探讨水环境中病毒携带新型耐药基因的情况及其耐药特点.通过分析病毒基因组的片段（reads）和重叠群（contigs），鉴定出多种耐药基因，包括AAC（6’）、Qnr A、Van Y、Vat和β-内酰胺酶基因.值得关注的是，通过核心序列比对和进化分析，挖掘出两种噬菌体携带的新型β-内酰胺酶基因bla_NSDPV-1和bla_NSVM-1.最小抑菌浓度试验表明，这两种编码新型β-内酰胺酶的耐药基因对临床常用头孢类和碳青霉烯类抗生素具有一定的耐药性.综上所述，水环境中噬菌体很可能在新型耐药基因的产生和耐药基因散播中起着重要的作用.另外，病毒宏基因组学结合体外实验是发现噬菌体携带新型耐药基因的重要方法.     

一株抗镉细菌的分离鉴定及其抗性基因定位的初步研究

报道了从长期重金属镉处理土壤中分离纯化 ,获得一株可在含镉 2 0 0mg L的培养基上正常生长的抗镉细菌菌株 ,编号为DKC1.经对该菌株进行形态观察、Biolog生理生化分析、G Cmol%测定及 16SrDNA序列同源性比较 ,该菌株鉴定为Ral stoniaeutropha .利用RDPⅡ数据库的PhylipInterface进行了系统发育地位分析 ,构建了进化树 ,并对KDC1菌株进行了抗重金属谱测定 ,结果表明该菌株对镉有较高的抗性 (35 0mg L)而对其它重金属耐性较低 ,通过质粒检测 ,该菌株含有一较大质粒 ,抗镉基因位于质粒上 .     

土壤中抗生素抗性基因的分布及迁移转化

为明晰土壤中抗生素抗性基因的存在及传播状况,并为开展抗生素抗性基因土壤污染的风险评价工作提供科学的理论依据.本文根据抗生素抗性基因存在和转移的理论基础,结合国内外研究进展,详细阐述了土壤中抗生素抗性基因的来源、分布传播及其影响因素,发现已有研究对外源抗生素抗性基因在土壤中的丰度和种类变化的研究较为深入,而对其在土壤中的空间分布特征、传播扩散及行为机理等研究工作尚处于起步阶段.因此提出未来土壤抗生素抗性基因的研究重点应为掌握抗生素抗性基因的空间分布、溯源、迁移转化规律及其关键制约因子,并建立相应的数量关系.     

海洋耐镉微生物筛选及其生物学特性

从重金属污染的近海域筛选到一株对镉具有较强抗性和富集能力的微生物Cd20002,该菌株在液体培养基中的最大耐受浓度为1 000 mg/L,在镉浓度达到200 mg/L时,其吸附率高达93.4%,根据形态特征和生理生化鉴定及16S rRNA测序鉴定为蓝杆菌。通过在不同的镉浓度环境对蓝杆菌Cd20002进行培养,测定其生长曲线,并优化菌体生长的条件及其吸附能力。结果表明,蓝杆菌Cd20002既能在重金属镉环境中生存,且富镉能力较强,对水产环境中重金属的生物清排具有重要意义。     

水产养殖环境中抗生素抗性基因的研究进展

为摸清我国水产养殖环境抗生素抗性基因（Antibiotic resistance genes,ARGs）污染状况,统计了当前水产养殖环境水、沉积物和养殖对象中七大类抗性基因的赋存状况,进一步分析了影响其赋存的因素,对未来我国水产养殖ARGs污染研究进行了展望.水产养殖环境中ARGs主要以iDNA形式存在;磺胺类和四环素类抗性基因的丰度和检出率较高,被认为是水产养殖环境主要的ARGs,已受到当前研究的广泛关注;而喹诺酮类、氯霉素类、β-内酰胺类、大环内酯类和氨基糖苷类抗性基因报道的丰度和检出率相对较低.养殖过程中,养殖用药、饲料及养殖模式是影响水产养殖环境ARGs赋存的主要因素,但环境理化因子调节养殖环境ARGs的作用也不容忽视.