纳污河流抗性基因和微生物群落相关性

通过Miseq高通量测序分析和荧光定量PCR技术,研究某纳污河流中四环素类与磺胺类抗性基因（ARGs）的分布特征、传播情况及微生物群落结构相关性.结果表明：在河流地表水与沉积物中均检测到四环素类抗性基因tetA和tetB与磺胺类抗性基因sul1和sul2,四环素类抗性基因和磺胺类抗性基因的绝对丰度分别为5.09×10³~1.26×10⁷,3.94×10⁵~1.32×10⁹copics/L,磺胺sul1抗生素抗性基因丰度显著高于其它基因;河流主要优势菌门为Proteobacteria,Bacteroidetes,Firmicutes,Actinobacteria和Cyanobacteria,其平均总相对丰度占总比例的95.62%,且总体差异较小.抗性基因与微生物群落冗余分析显示,Methylotenera菌属是影响tetA抗性基因分布丰度的主要因素,Dechloromonas和Clostridium sensu stricto 1菌属是影响tetB抗性基因分布丰度的主要因素,Dechloromonas、Clostridium sensu stricto 1和Methylotenera菌属是影响sul1、sul2抗性基因分布丰度的主要因素.     

污水处理厂中磺胺类抗生素、抗性菌、抗性基因的特性

抗生素滥用引起抗性菌的扩散和抗生素抗性基因的污染已成为严重的公共卫生安全隐患。从某污水处理厂出水中分离磺胺类抗性菌,检测其对抗生素耐受性及磺胺类抗生素浓度、抗性基因(antibiotic resistance genes,ARGs)的含量。结果表明水中磺胺类抗生素检测出5种磺胺类抗生素,但浓度较低,3种磺胺类抗生素未检出;磺胺类抗性菌含量为3.18×102~2.24×104CFU/m L,分离的抗性菌对9种抗生素具有抗性,最高耐受力为512 mg/m L,其中,对氯霉素耐受能力最强,对青霉素、氨苄青霉素、头孢氨苄、环丙沙星、庆大霉素、阿奇霉素耐受力次之,对利福平耐受力最弱;抗性基因的绝对含量较低,其中最高含量sul1为105.648~106.956。该研究为STPs抗生素污染的风险评估与抗生素抗性基因污染控制提供了基础数据支持。     

江苏省代表性水源地抗生素及抗性基因赋存现状

抗生素和抗生素抗性基因（antibiotic resistance genes,ARGs）是环境中重要的新兴污染物,为探明江苏省代表性水源地多种环境介质中抗生素和ARGs的污染水平及影响因素,于2018年12月和2019年6月采集苏北、苏中和苏南的5处代表性集中式饮用水水源地取水口处水体、表层沉积物和石相附着生物膜样品,对3种介质中10种代表性抗生素浓度、1类整合子酶基因intl1和7种代表性ARGs的绝对丰度进行检测分析.结果表明,5处水源地中目标抗生素和ARGs处于较低赋存水平.磺胺类抗生素在水体、表层沉积物和附着生物膜中的赋存量分别为NF（未检出）~37.4 ng·L^-1,NF~47.3 ng·g^-1和NF~3759.1 ng·g^-1.喹诺酮类抗生素在3种介质中的浓度和含量分别为NF~5.3 ng·L^-1、0.4~32.5 ng·g^-1和NF~4220.9 ng·g^-1.目的ARGs中,sul1、sul2、tetW和tetQ的检出率为100%,其中磺胺类抗生素ARGs,即sul1和sul2基因丰度最高.表层沉积物和附着生物膜中的ARGs丰度相当,高于水体中ARGs的丰度.网络分析结果表明,所属拟杆菌门、变形菌门、厚壁菌门、疣微菌门和放线菌门的细菌最有可能成为代表性水源地中ARGs的潜在宿主,在ARGs的扩散和转移过程中起重要作用.研究结果对于江苏省集中式饮用水源地水环境质量状况评估和水质安全保障具有一定的科学指导意义.     

北京地区菜田土壤抗生素抗性基因的分布特征

为研究北京地区菜田土壤抗生素抗性基因(antibiotic resistance genes,ARGs)的污染状况和分布特征,采集了11个长期施用粪肥蔬菜基地的温室土壤和大田土壤,进行了抗生素以及ARGs种类和丰度的检测.结果表明,菜田土壤中四环素类抗生素残留量最高,其次为磺胺类抗生素,温室土壤抗生素残留普遍高于大田土壤.温室和大田土壤磺胺类抗性基因sul1、sul2和四环素类抗性基因tet L的检出率均为100%.温室土壤的Ⅰ类整合子(int I1)检出率比大田土壤高1.5倍.定量PCR的检测结果表明,菜田土壤中sul2和tet L的相对丰度介于10-4~10-2之间,温室土壤sul2和tet L的相对丰度普遍高于大田土壤.sul2的相对丰度与磺胺二甲嘧啶和强力霉素的含量显著正相关(P0.05),tet L相对丰度与抗生素含量无明显相关性(P0.05).本研究结果对于掌握北京地区农田土壤ARGs的污染现状,以及从ARGs角度评估农艺措施具有重要的指导意义.     

珠江口典型水产养殖区抗生素抗性基因污染的初步研究

使用普通PCR和荧光定量PCR方法对珠江口典型水产养殖区水和沉积物中3种磺胺类、7种四环素类、1种喹诺酮类抗生素抗性基因(antibiotic resistance genes,ARGs)和1种整合子基因进行了定性和定量研究.结果表明,除tetW外,所有其他ARGs在珠江口养殖区中均被检出,其中sul1、sul2和int1的检出率为100%.相同养殖模式下,养殖时间越长ARGs的相对含量越高,表明ARGs具有累积效应;不同养殖模式池塘中ARGs的含量存在差异,表明养殖模式可能影响ARGs的含量与分布.int1相对含量与sul1和ARGs总量之间均存在显著相关性(P<0.05),表明int1在ARGs的水平传播中起着非常重要的作用.此外,珠江口水产养殖区沉积物中抗生素浓度与ARGs总量存在显著相关性(P<0.05),说明水产养殖区中抗生素残留是诱导养殖区ARGs的主要因素.     

一株磺胺二甲嘧啶抗性菌的筛选及抗性机制研究

为研究地下水中磺胺类抗性菌的抗性特征与抗性机制,在调研北京市东南发展区地下水受抗生素污染情况基础上,从中筛得一株磺胺二甲嘧啶（SMZ）抗性菌SMZ-R9,经形态学观察、生理生化试验及16S rRNA基因测序分析,鉴定为乳酸不动杆菌（Acinetobacter lactucae）.使用荧光定量PCR与普通PCR定量以及定性检测抗性基因（ARGs）,发现菌株SMZ-R9同时携带3种磺胺ARGs（sul1、sul2、sul3）、1种甲氧苄啶ARGs（dfrA1）以及Int1基因;通过对基因序列进行同源性比对及KEGG功能注释,发现sul2、dfrA1基因为编码二氢蝶呤合成酶（DHPs）与二氢叶酸还原酶（DHFR）的核酸序列,表达产物分别催化叶酸合成的上游与下游途径,共同介导菌株SMZ-R9的SAs抗性.可见,通过对ARGs编码蛋白的功能注释与分析,可为微生物的抗生素抗性机制在基因水平的研究提供理论依据.      

长江口近岸地区抗生素抗性基因与微生物群落分布特征

由于抗生素的大量使用,环境中微生物对抗生素的抗性不断增加,抗生素抗性基因(ARGs)问题越来越严重,严重威胁生态安全和人类健康.为研究长江口近岸地区水体和底泥沉积物中的ARGs和微生物群落的分布特征,通过野外调查采集了8个站点的水样和沉积物样本,对2种磺胺类抗性基因(sul1、sul2)、6种四环素类抗性基因(tetM、tetC、tetX、tetA、tetO、tetQ)、1种整合子基因intI1、16S rRNA基因和微生物群落进行检测分析.结果表明,长江口近岸地区10种抗性基因的检出率为100%.其中,整合子基因intI1和水样中多种ARGs呈显著正相关关系.变形菌门(Proteobacteria)和拟杆菌门(Bacteroidota)是长江口近岸地区水环境中的优势菌门;Chloroplast为水体中的主要菌属,Chloroplast和Nitrospira为沉积物中的主要菌属.在水体中,硝化螺旋菌门(Nitrospirota)是4种四环素类抗性基因(tetX、tetA、tetO和tetQ)共同的潜在宿主;在沉积物中,Sva0485是sul1和intI1的共同潜在宿主.微生物群落的分布...     

人工湿地去除畜禽养殖废水中磺胺类抗生素抗性基因研究

采用垂直潜流人工湿地研究了畜禽养殖废水中磺胺类抗生素抗性基因(ARGs)在人工湿地中的去除及累积情况.结果表明,畜禽养殖废水中的3种磺胺类ARGs(sul Ⅰ,sul Ⅱ及sul Ⅲ)的平均绝对含量分别为1.15×1010、7.51×1010及7.51×107 copies/L.通过湿地系统处理后sul Ⅰ、sul Ⅱ及sul Ⅲ的平均去除率分别为89％、88％及84％.在系统运行末期,湿地表层土壤和底层土壤中sul Ⅰ、sul Ⅱ及sul Ⅲ的绝对拷贝数和相对表达量均有明显的升高.结果表明,人工湿地系统可有效降低畜禽养殖废水中ARGs含量.     

同一城市的两个市政污水处理厂中抗生素抗性基因消长研究

为了考察处理工艺对污水中抗生素抗性基因(antibiotic resistance genes,ARGs)削减的影响,本研究检测了同一城市的两个市政污水处理厂(1#:缺氧+好氧,2#:CASS+纤维转盘滤池)污水中六种典型ARGs(tetX、sul1、blaTEM、qnrS、ermB、intl1)绝对丰度和相对丰度的变...     

长江武汉段水源地典型抗生素及抗性基因污染特征与生态风险评价

为了探究长江武汉段水源地抗生素和抗性基因(ARGs)的分布特征和相关性以及潜在生态风险,采用固相萃取-超高效液相色谱串联质谱法(SPE-UPLC-MS/MS)和实时荧光定量PCR技术分别分析了全武汉市饮用水水源地中13种抗生素和10种ARGs的赋存水平.结果表明,在16处水源地样品中共检出9种抗生素,浓度范围为ND(未检出)～177.36 ng·L^-1,红霉素、磺胺嘧啶和磺胺对甲氧嘧啶的检出率均为100%,抗生素浓度分布呈现(以汉江汇入口划分长江干流上下游):支流举水<长江干流下游<长江干流上游<支流汉江<支流滠水.长江干流下游ARGs总绝对丰度明显高于长江干流上游和汉江,磺胺类ARGs显著高于其他三类ARGs耐药基因的平均丰度(P<0.05),ARGs中sul1与sul2、ermB、qnrS、tetW和intI1等5种ARGs存在显著正相关关系(P<0.01),相关系数分别为0.768、 0.648、 0.824、 0.678和0.790,磺胺类ARGs组内相关性弱于组间ARGs的相关性.磺胺甲■唑、金霉素、罗红霉素和恩诺沙星...     

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.     

Arsenic uptake by arbuscular mycorrhizal maize （Zea mays L.） grown in an arsenic-contaminated soil with added phosphorus

The effects of arbuscular mycorrhizal (AM) fungus (Glomus mosseae) and phosphorus (P) addition (100 mg/kg soil) on arsenic (As) uptake by maize plants (Zea mays L.) from an As-contaminated soil were examined in a glasshouse experiment.Non-mycorrhizal and zero-P addition controls were included.Plant biomass and concentrations and uptake of As,P,and other nutrients,AM colonization,root lengths,and hyphal length densities were determined.The results indicated that addition of P significantly inhibited root colonization and development of extraradical mycelium.Root length and dry weight both increased markedly with mycorrhizal colonization under the zero-P treatments,but shoot and root biomass of AM plants was depressed by P application.AM fungal inoculation decreased shoot As concentrations when no P was added,and shoot and root As concentrations of AM plants increased 2.6 and 1.4 times with P addition,respectively.Shoot and root uptake of P,Mn,Cu,and Zn increased,but shoot Fe uptake decreased by 44.6%,with inoculation, when P was added.P addition reduced shoot P,Fe,Mn,Cu,and Zn uptake of AM plants,but increased root Fe and Mn uptake of the nonmycorrhizal ones.AM colonization therefore appeared to enhance plant tolerance to As in low P soil,and have some potential for the phytostabilization of As-contaminated soil,however,P application may introduce additional environmental risk by increasing soil As mobility.     

Degradation of metabolites accumulated of benzo[a]pyrene by coupling Penicillium Chrysogenum with KMnO4

Several main metabolites of benzo[a]pyrene （BaP） formed by Penicillium chrysogenum, Benzo[a]pyrene-1,6-quinone （BP 1,6- quinone）, trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene （BP 7,8-diol）, 3-hydroxybenzo[a]pyrene （3-OHBP）, were identified by high-performance liquid chromatography （HPLC）. The three metabolites were liable to be accumulated and were hardly further metabolized because of their toxicity to microorganisms. However, their further degradation was essential for the complete degradation of BaP. To enhance their degradation, two methods, degradation by coupling Penicillium chrysogenum with KMnO4 and degradation only by Penicillium chrysogenum, were compared; Meanwhile, the parameters of degradation in the superior method were optimized. The results showed that （1） the method of coupling Penicillium chrysogenum with KMnO4 was better and was the first method to be used in the degradation of BaP and its metabolites; （2） the metabolite, BP 1,6-quinone was the most liable to be accumulated in pure cultures; （3） the effect of degradation was the best when the concentration of KMnO4 in the cultures was 0.01% （w/v）, concentration of the three compounds was 5 mg/L and pH was 6.2. Based on the experimental results, a novel concept with regard to the bioremediation of BaP-contaminated environment was discussed, considering the influence on environmental toxicity of the accumulated metabolites.     

Impact of atrazine and nitrogen fertilizers on the sorption of chlorotoluron in soil and model sorbents

Sorption of chlorotoluron in ammonium sulfate, urea and atrazine multi-solutes system was investigated by batch experiments. The results showed application of nitrogen fertilizers to the soil could affect the behavior of chlorotoluron. At the same concentration of N, sorption of chlorotoluron decreased as the concentration of atrazine increased on the day 0 and 6 in soil, respectively. The sorption of chlorotoluron increased from 0 to 6 d when soils were preincubated with deionized water, ammonium sulfate and urea solution for 6 d. That indicated incubation time was one of the most important factors for the sorption of chlorotoluron in nitrogen fertilizers treatments. The individual sorption isotherms of chlorotoluron in rubbery polymer and silica were strictly linear in single solute system, but there were competition sorption between pesticides or between pesticides and nitrogen fertilizers. That indicated the sorption taken place by concurrent solid-phase dissolution mechanism and sorption on the interface of water-organic matter or water-mineral matter.     

Distribution and ecological effect of mercury in Laogang landfill, Shanghai, China

Laogang landfill near Shanghai is the largest landfill in China, and receives about 10000 t of daily garbage per day, Samples of topsoil and plants were analyzed to evaluate mercury pollution from the landfill. For topsoil samples, there were significant correlations among total mercury （HgT）, combinative mercury （Hgc） and gaseous mercury （HgG）, and content of total organic carbon （TOC）, but, no significantly relationship was found between Hg content and filling time. Hg content changes in vertical profiles with time showed that the average Hgv of profiles 1992, 1996, and 2000 was similar, but their average HgG was quite different. HgT was significantly correlated with Hgc in profile 1992 and 2000, and Hgv was significantly correlated with Hg6 in profile 1996. HgG/Hgv ratio in profile samples decreased in the order of （HgG,/HgT）1992〉（HgG/HgT）1996〉〉（HgG/HgT）2000. A simple outline of Hg release in landfill could be drawn： with increasing of filling time, degradation undergoes different biodegradation, accordingly, gaseous mercury goes through small, more, and small proportion to total mercury. Distribution of Hg in plants was inhomogeneous, following the order of leaf〉root〉stem. The highest value of leaf may be associated with higher atmospheric Hg from landfill. Ligneous plants （e.g. Phyllostachys glanca, Prunus salicina and Ligustrum lucidum） are capable of enriching more Hg than herbaceous plants.     

EDTA-enhanced phytoremediation of lead contaminated soil by Bidens maximowicziana

Phytoremediation is a potential cleanup technology for the removal of heavy metals from contaminated soils.Bidens maximowicziana is a new Pb hyperaccumulator,which not only has remarkable tolerance to Pb but also extraordinary accumulation capacity for Pb.The maximum Pb concentration was 1509.3 mg/kg in roots and 2164.7 mg/kg in overground tissues.The Pb distribution order in the B. maximowicziana was:leaf>stem>root.The effect of amendments on phytoremediation was also studied.The mobility of soil Pb and the Pb concentrations in plants were both increased by EDTA application.Compared with CK(control check),EDTA application promoted translocation of Pb to overground parts of the plant.The Pb concentrations in overground parts of plants was increased from 24.23-680.56 mg/kg to 29.07-1905.57 mg/kg.This research demonstrated that B.maximowicziana appeared to be suitable for phytoremediation of Pb contaminated soil,especially,combination with EDTA.     

Decomposition of alachlor by ozonation and its mechanism

Decomposition and corresponding mechanism of alachlor, an endocrine disruptor in water by ozonation were investigated. Results showed that alachlor could not be completely mineralized by ozone alone. Many intermediates and final products were formed during the process, including aromatic compounds, aliphatic carboxylic acids, and inorganic ions. In evoluting these products, some of them with weak polarity were qualitatively identified by GC-MS. The information of inorganic ions suggested that the dechlorination was the first and the fastest step in the ozonation of alachlor.     

Enzymatic oxidation of aqueous pentachlorophenol

The influence of the nonionic surfactant Tween 80 on pentachlorophenol （PCP） oxidation catalyzed by horseradish peroxidase was studied. The surfactant was tested at concentrations below and above its critical micelle concentration （CMC）. Enhancement of PCP removal was observed at sub-CMCs. The presence of Tween 80 in the reaction mixture reduced enzyme inactivation which occurred through a combination of free radical attack and sorption by precipitated products. A simple first-order model was able to simulate time profiles for enzyme inactivation in the presence or absence of Tween 80. At supra-CMCs, the surfactant caused noticeable reductions in PCP removal, presumably through micelle partitioning of PCP which precluded the hydrophobic PCP molecule from interacting with the enzyme.     

Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes

In this study an effort has been made to use plant polyphenol oxidases; potato （Solanum tuberosum） and brinjal （Solanum melongena）, for the treatment of various important dyes used in textile and other industries. The ammonium sulphate fractionated enzyme preparations were used to treat a number of dyes under various experimental conditions. Majority of the treated dyes were maximally decolorized at pH 3.0. Some of the dyes were quickly decolorized whereas others were marginally decolorized. The initial first hour was sufficient for the maximum decolorization of dyes. The rate of decolorization was quite slow on long treatment of dyes. Enhancement in the dye decolorization was noticed on increasing the concentration of enzymes. The complex mixtures of dyes were treated with both preparations of polyphenol oxidases in the buffers of varying pH values. Potato polyphenol oxidase was significantly more effective in decolorizing the dyes to higher extent as compared to the enzyme obtained from brinjal polyphenol oxidase. Decolorization of dyes and their mixtures, followed by the formation of an insoluble precipitate, which could be easily removed simply by centrifugation.     

Removal of heavy metals from sewage sludge by low costing chemical method and recycling in agriculture

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