市售牛奶中4种四环素类抗生素抗性基因的比较研究

目的研究市售牛奶商品中4种代表性四环素类抗生素抗性基因(antibiotic resistance genes,ARGs)的分布特征。方法以12种商品牛奶为研究对象,通过对样品中的4种四环素类ARGs和16S r RNA进行实时荧光定量PCR测定,分析ARGs的分布特征,并对样品的保存条件、保质期、成分含量做相关性分析。结果牛奶样品中普遍检出抗性基因Tet(A)、Tet(W),检出率为100%,其抗性机制分别是外排泵和细胞保护;Tet(32 s)和Tet(L)均未检出。比较样品中ARGs的相对丰度,Tet(A)最高达到2.28,最低为0.06;Tet(W)最高达到0.24,最低为0.03。牛奶的产品类型、包装形式、保存条件和保质期与Tet(W)的相对丰度显著相关。结论市售牛奶样品中存在ARGs,不同牛奶样品中ARGs的富集程度存在差异。牛奶样品中ARGs相对丰度受到多种因素影响,值得从食品安全的角度深入研究。     

水产品批发市场中抗生素抗性基因的污染分析

为了探究待售水产品中抗生素抗性基因(Antibiotic resistance genes,ARGs)在水产品批发市场中的扩散污染情况,选取上海市3个大型水产品批发市场作为研究对象,采用PCR的方法,对市场内部和市场附近的排污沟表层淤泥中5大类(4种磺胺类、8种四环素类、6种β-内酰胺类、5种氯霉素类、4种链霉素类)共27种ARGs进行了检测,并利用PCR-DGGE技术分析了细菌种群多样性。结果显示共有17种ARGs在批发市场淤泥中被检出,其中磺胺类、四环素类和链霉素类ARGs的检出率较高且检出种类较多,表明水产品批发市场含有种类丰富的ARGs,水产品以磺胺类、四环素类和链霉素类ARGs污染为主。对市场外围的检测显示市场外围区域ARGs的种类分布略低于市场内部,表明水产品批发市场中ARGs的存在会通过污水排放途径扩散到周边环境。DGGE图谱的分析结果显示,3个批发市场污泥样品细菌种类差异性较大,表明环境中ARGs的种类分布与细菌的种类并无直接关联。本研究表明水产品携带的ARGs会扩散到水产品批发市场中,导致水产品批发市场成为ARGs的重要储存库,以此造成的污染会进一步传播到周边环境,此外,这些种类丰富的ARGs重新扩散到待售水产品中的风险很大,需引起人们足够的重视。     

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_TEM82.29%)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...     

4种市售水产品抗生素耐药基因和intⅠ1实时荧光定量PCR检测

为掌握水产品中抗生素耐药基因(ARGs)和Ⅰ类整合子intⅠ1含量及分布,本研究建立了水产品常见抗生素耐药基因实时荧光定量PCR(qPCR)检测体系。以含目的基因tetA、sul2、blaPSE、cmlA、qnrS、aac(6')-Ib、intⅠ1的重组质粒为标准品,建立qPCR标准曲线,按照已建立的qPCR体系对市售沼虾、鲫鱼、黄颡鱼、大黄鱼中ARGs和intⅠ1进行定量。结果表明,6种ARGs的最低检出限在11.5～202拷贝/μL之间,blaPSE的最低检出限为6.6×10~4拷贝/μL。沼虾、鲫鱼、黄颡鱼、大黄鱼4种样品都存在高丰度的blaPSE,相对丰度范围为1.17×10~(-4)～5.96×10~(-3),aac(6')-Ib的相对丰度在4种样品中都较低,在2.38×10~(-8)～6.71×10~(-7)之间。该检测体系以样品中细菌总基因组为模板,可对水产品中6种常见ARGs同时定量检测,能够较客观地揭示市售水产样品中ARGs的种类和数量分布信息,对控制水产品中ARGs污染并保障水产食品安全意义深远。     

南美白对虾肠道内乳酸菌的分离鉴定及其对抗生素的耐药性

以南美白对虾为研究对象,采用平板分离法从其肠道内筛得26株乳酸菌,通过16S rRNA测序鉴定可归为4种:乳酸乳球菌乳亚种L. lactis subsp. lactis、台湾乳球菌L. taiwanensis、格式乳球菌L. garvieae、乳酸乳球菌L. lactis。选取7株具有代表性的乳酸菌,用PCR的方法检测对虾中检出率较高的6类18种抗性基因(ARGs)在乳酸菌中的分布,辅以平板涂布的方法用抗生素选择性培养基研究乳酸菌对抗生素的耐药性。研究发现:7株乳酸菌具有相似的耐药谱,对四环素、磺胺吡啶、乙酰螺旋霉素表现出耐药性(抑菌率50.00%),对盐酸金霉、土霉素、硫酸庆大霉素、氯霉素、红霉素敏感(抑菌率100%);含有多重抗性基因,ARGs的检出率:磺胺类(92.86%)四环素类(53.06%)喹诺酮类(23.81%)氨基糖苷类(4.76%)氯霉素类=大环内酯类(0.00%),其ARGs基因型与菌株的抗生素表型并不能完全吻合。     

基于SmartChip HT-qPCR分析不同食品中抗生素抗性基因的多样性及丰度

抗生素抗性基因（Antibiotics Resistance Genes,ARGs）广泛存在于各种环境中,被认为是一种新型环境污染物,对环境及人类健康具有潜在威胁。但是目前对于不同食品中多种ARGs赋存状况的研究仍然有限。该研究采用SmartChip高通量实时定量PCR方法（High-Throughput Quantitative Real-Time PCR,HT-qPCR）对水产品、发酵食品、肉类、蔬菜四大类样品中的ARGs及可移动遗传元件（Mobile Genetic Elements,MGEs）进行了定量分析。结果显示,被分析的15个样品中共检出9个ARGs大类,涉及234个ARGs亚类。所有样品中共有的ARGs亚类为79个,特有的为51个;水产品和肉类样品富集多种ARGs,这些基因表现出较高的丰度（最高可达3.6 copies/16S rRNA gene）。MGEs的检出数量达10个,其中所有样均检出整合子基因intI-1(clinic)和转座酶基因tnpA-01。ARGs抵抗机制分析结果表明,抗生素灭活机制、外排泵机制和细胞保护机制为主要的ARGs抵抗机制,抵抗机制贡献大小依次为：抗生素灭活机制>外排泵机制>细胞保护机制。该研究进一步丰富了对不同食品中ARGs赋存情况的认识,可为食品中ARGs的风险分析提供依据。     

四川广元植烟土壤微塑料分布状况分析

【目的】为明确植烟土壤微塑料分布特征，解决微塑料污染的环境问题。【方法】以四川省广元市剑阁县汉阳镇植烟土壤为研究对象，采集25个调查点的土壤样品，采用密度浮选法提取土壤微塑料，通过体式显微镜观察土壤样品中微塑料丰度、面积、颜色、类型和粒径。【结果】广元烟田土壤微塑料丰度范围在1.20×10²～8.9×10² pieces·kg^-1，平均丰度为(4.81±3.53)×10² pieces·kg^-1，平均面积为(4.45±4.98)×10² mm²·kg^-1；土壤中主要存在4种类型的微塑料，分别是薄膜、碎片、泡沫、纤维，占比最多的为薄膜类；颜色存在黑色和透明，绝大部分以透明状态存在。薄膜、碎片、泡沫、纤维类微塑料的粒径平均值分别为1.18、1.12、1.69和2.14 mm。土壤微塑料粒径为[0-0.5)、[0.5-1)、[1-2)和[2-5) mm的分别占微塑料总数的5.59%、28.28%、48.41%和17.7...     

卷烟主流烟气气溶胶分布研究

对卷烟主流烟气气溶胶分布进行了研究,按照标准吸烟条件,对4个卷烟进行了测试,通过在线200倍稀释,采用测量范围在7～10000 nm的电子低压撞击器对气溶胶粒径和浓度进行了测试。试验结果表明,不同卷烟主流烟气气溶胶分布有一定的差异,烟气气溶胶浓度随抽吸口数增加而增加,所测试的A、D样品气溶胶最高浓度粒径出现在0.261～0.381μm,B、C样品卷烟气溶胶最高浓度粒径出现在0.028μm;A、D样品卷烟气溶胶平均粒径为0.23μm,而B、C样品卷烟平均粒径为0.16μm;4种卷烟主流烟气气溶胶平均浓度介于1.09×109～2.21×109个/cm3之间,而相应每支烟的总粒子数介于2.29×1011～3.88×1011个粒子。     

基于人体吸入和呼出烟气气溶胶粒径分布的气溶胶呼吸道沉降研究

为了考察烟气气溶胶在人体呼吸道的沉降特征,采用吸烟循环模拟机-快速粒径谱仪(SCS-DMS),基于志愿者实际抽吸曲线,对人体抽吸卷烟、加热卷烟和电子烟的吸入和呼出烟气气溶胶粒径分布进行在线表征,并采用多路径粒子剂量模型(MPPD),模拟研究了3类烟草制品的烟气气溶胶在人体呼吸道的沉降情况。结果表明:①在志愿者吸入和呼出过程中,卷烟、加热卷烟和电子烟烟气气溶胶的粒径增长因子分别为1.25~1.40、1.52~2.03和1.44~2.43。②不同类型样品的烟气气溶胶在呼吸道各部位的沉降系数为卷烟＜加热卷烟＜电子烟。③3类样品的烟气气溶胶在人体呼吸道不同部位的沉降系数均为口腔 < 气管支气管 < 肺部。④在右下肺叶的沉降系数相对较高,在肺部的最大沉降系数出现在第19~22级气管。      

电加热卷烟气溶胶物理特性的表征

收集市场上5个品牌的20个电加热卷烟样品,对其总释放气溶胶和逐口释放气溶胶的物理特性(粒数浓度、粒数中值粒径和体积浓度)进行表征及对比分析.结果表明:1)电加热卷烟样品总释放气溶胶的粒数浓度主要在109个/cm3的量级,体积浓度在105μm3/cm3的量级,粒径分布轮廓为单峰分布或双峰分布,粒数中值粒径主要集中在30～...     

Prevalence of clinically relevant antibiotic resistance genes in surface water samples collected from Germany and australia

The prevalence and proliferation of antibiotic resistant bacteria is profoundly important to human health, but the extent to which aquatic environments contribute toward the dissemination of antibiotic resistant genes (ARGs) is poorly understood. The prevalence of 24 ARGs active against eight antibiotic classes (β-lactams, aminoglycosides, glycopeptides, chloramphenicols, tetracycline, macrolides, trimethoprim, and sulfonamides) was evaluated in surface water samples collected from Germany and Australia with culture independent methods. The ARGs most frequently detected both in Germany and Australia were sulI, sulII (77-100%), and dfrA1 (43-55%) which code for resistance to sulfonamide and trimethoprim. Macrolides resistance gene ermB was relatively more prevalent in the surface water from Germany (68%) than Australia (18%). In contrast, the chloramphenicol resistance gene catII was more frequently detected in Australia (64%) than Germany (9%). Similarly, β-lactams resistance gene ampC was more prevalent in the samples from Australia (36%) than Germany (19%). This study highlights wide distribution of ARGs for sulfonamide, trimethoprim, macroline, β-lactams and chloramphenicol in the aquatic ecosystems. Aquatic ecosystems can therefore be reservoirs of ARGs genes which could potentially be transferred from commensal microorganisms to human pathogens.     

Antibiotic resistance genes as emerging contaminants: studies in northern Colorado

This study explores antibiotic resistance genes (ARGs) as emerging environmental contaminants. The purpose of this study was to investigate the occurrence of ARGs in various environmental compartments in northern Colorado, including Cache La Poudre (Poudre) River sediments, irrigation ditches, dairy lagoons, and the effluents of wastewater recycling and drinking water treatment plants. Additionally, ARG concentrations in the Poudre River sediments were analyzed at three time points at five sites with varying levels of urban/agricultural impact and compared with two previously published time points. It was expected that ARG concentrations would be significantly higher in environments directly impacted by urban/agricultural activity than in pristine and lesser-impacted environments. Polymerase chain reaction (PCR) detection assays were applied to detect the presence/absence of several tetracycline and sulfonamide ARGs. Quantitative real-time PCR was used to further quantify two tetracycline ARGs (tet(W) and tet(O)) and two sulfonamide ARGs (sul(I) and sul(II)). The following trend was observed with respect to ARG concentrations (normalized to eubacterial 16S rRNA genes): dairy lagoon water > irrigation ditch water > urban/agriculturally impacted river sediments (p < 0.0001), except for sul(II), which was absent in ditch water. It was noted that tet(W) and tet(O) were also present in treated drinking water and recycled wastewater, suggesting that these are potential pathways for the spread of ARGs to and from humans. On the basis of this study, there is a need for environmental scientists and engineers to help address the issue of the spread of ARGs in the environment.     

Sources of fine particles in a rural midwestern U.S. area

Ambient PM2.5 (particulate matter < or = 2.5 microm in aerodynamic diameter) samples collected at a rural monitoring site in Bondville, IL on every third day using Interagency Monitoring of Protected Visual Environments (IMPROVE) sampler were analyzed through the application of the positive matrix factorization (PMF). The particulate carbon fractions were obtained from the thermal optical reflectance method that divides particulate carbon into four organic carbon, pyrolyzed organic carbon (OP), and three elemental carbon fractions. A total of 257 samples collected between March 2001 and May 2003 analyzed for 35 species were used and eight sources were identified: summer-high secondary sulfate aerosol (40%), secondary nitrate aerosol (32%), gasoline vehicle (9%), OP-high secondary sulfate aerosol (7%), selenium-high secondary sulfate aerosol (4%), airborne soil (4%), aged sea salt (2%), and diesel emissions (2%). The compositional profiles for gasoline vehicle and diesel emissions are similar to those estimated in other U.S. areas. Backward trajectories indicate that the highly elevated airborne soil impacts were likely caused by Asian and Saharan dust storms. Potential source contribution function analyses show the potential source areas and pathways of secondary sulfate aerosols, especially the regional influences of the biogenic as well as anthropogenic secondary aerosol.     

重金属和抗生素抗性基因在土壤中的复合污染研究进展

抗生素滥用及其诱导产生的抗生素抗性基因(antibiotic resistance genes, ARGs)污染己经引起人们的广泛关注。与此同时,环境中重金属的污染状况也呈加剧趋势,与抗生素抗性基因不同,重金属在自然环境中很难被降解,二者同时存在不仅维持和加强了环境中的微生物对重金属的原有抗性,同时增强了对抗生素抗性基因的抗性。目前,重金属污染地区的养殖业中动植物细菌传染病的抗生素治疗总是无效的,其原因可能是重金属离子和抗生素抗性基因的协同抗性在选择压力下得到增强。本文总结了我国抗生素抗性基因和重金属元素在土壤中的污染状况及其复合污染特征,综述了这2类物质在土壤中的协同选择机制和综合修复措施,以期为重金属和抗生素抗性基因带来的污染所致危害的减轻和未来复合污染相关研究的深入提供参考。     

Characterization of saccharides in size-fractionated ambient particulate matter and aerosol sources: the contribution of primary biological aerosol particles (PBAPs) and soil to ambient particulate matter

Size-fractionated (equivalent to ambient PM2.5 and PM10) local soil, plant, and spore samples were collected in the Sonoran Desert near Phoenix, AZ and measured for saccharide content with the goal of characterizing ambient particulate matter sources including soil and primary biological aerosol particles (PBAPs) from plants and fungi. Different saccharide compositions were observed among soil, plant, and spore samples and between PM2.5 and PM10 fractions. The total measured nonlevoglucosan saccharide content relative to PM mass in ambient aerosols collected in a Phoenix suburb (Higley) was much higher compared to the local soil samples but much lower compared to the PBAP. The enrichment of saccharides from two saccharide-dominated PM source factors resolved by a positive matrix factorization model is also higher than the saccharide content in the size-fractionated local soil samples, but lower than that measured in the size-segregated PBAP samples. This indicates that ambient concentration of particulate saccharides at Higley was dominated by contributions from PBAPs directly injected into the atmosphere from plants and spores rather than from soil and associated biota. Our results also suggest the contribution to the fine size fraction of ambient PM from the primary biologically derived sources may be greater than previously acknowledged.     

Tertiary-treated municipal wastewater is a significant point source of antibiotic resistance genes into Duluth-Superior Harbor

In this study, the impact of tertiary-treated municipal wastewater on the quantity of several antibiotic resistance determinants in Duluth-Superior Harbor was investigated by collecting surface water and sediment samples from 13 locations in Duluth-Superior Harbor, the St. Louis River, and Lake Superior. Quantitative PCR (qPCR) was used to target three different genes encoding resistance to tetracycline (tet(A), tet(X), and tet(W)), the gene encoding the integrase of class 1 integrons (intI1), and total bacterial abundance (16S rRNA genes) as well as total and human fecal contamination levels (16S rRNA genes specific to the genus Bacteroides ). The quantities of tet(A), tet(X), tet(W), intI1, total Bacteroides , and human-specific Bacteroides were typically 20-fold higher in the tertiary-treated wastewater than in nearby surface water samples. In contrast, the quantities of these genes in the St. Louis River and Lake Superior were typically below detection. Analysis of sequences of tet(W) gene fragments from four different samples collected throughout the study site supported the conclusion that tertiary-treated municipal wastewater is a point source of resistance genes into Duluth-Superior Harbor. This study demonstrates that the discharge of exceptionally treated municipal wastewater can have a statistically significant effect on the quantities of antibiotic resistance genes in otherwise pristine surface waters.     

Occurrence, abundance, and diversity of tetracycline resistance genes in 15 sewage treatment plants across China and other global locations

Activated sludge was sampled from 15 sewage treatment plants (STPs) across China and other global locations to investigate the occurrence, abundance and diversity of tetracycline resistance genes (tet) in the STPs. Occurrence and abundance of 14 tet genes were determined using polymerase chain reaction (PCR) and quantitative real time PCR. Six genes (tet(A), tet(C), tet(G), tet(M), tet(S), and tet(X)) were detected in all the STPs, while no sludge sample contained tet(Q). Total concentration of the 14 genes was significantly different among the STPs and average tet abundance of the STPs varied greatly among the tet types (p<0.05). Tet(G) had the highest concentration in the STPs, followed by tet(C), tet(A) and tet(S). Phylogenetic diversity of the genes was investigated using DNA cloning. BLAST analysis showed that all of the 450 cloned sequences matched known tet genes, except for tet(G). The 56 tet(G) clones were grouped into 14 genotypes, among which type G24 had an identical sequence to tet(G) carried by Salmonella enterica or Acinetobacter baumannii, while the other sequences had low similarity to the known genes in GenBank. The results of this study might be useful to understand the diversity of these resistance genes in STPs.