噬菌体编码的抗生素抗性基因研究进展

噬菌体作为地球上丰度最高的生物体,对于全球生态系统的微生物群落组成与动态,生物地球化学循环和细菌进化等都具有重要作用,但噬菌体对抗生素抗性的贡献还所知甚少.通过宏基因组测序和定量PCR等技术,发现了海洋,河流,土壤,动物肠道等环境的噬菌体基因组中存在多种耐药基因(antibiotic resistance genes,ARGs).因此,噬菌体及其介导的转导作用可能对ARGs的富集扩散发挥着重要作用.本文综述了不同环境介质的噬菌体基因组中ARGs的分布特征以及噬菌体介导的ARGs水平转移的研究进展,同时展望了为阐明噬菌体对抗生素抗性贡献需要解决的科学问题.本文将为了解噬菌体在ARGs传播扩散中的作用以及扩展对可移动抗性组的认知奠定基础.     

长江中上游部分江段水细菌质粒的研究

<正> 质粒是宿主细胞染色体外的遗传因子,它常常携带着有重要功能的基因,据报道:细菌的产毒性、抗药性以及对难分解物质的降解性与质粒都有密切关系。许多研究表明,虽然质粒较广泛地分布在细菌各种属中,但在一般环境中它对宿主本身的生存并非都是必要的。在不良条件下,如营养物质缺乏,有毒污染物和酸、碱等限制因子存在的情况下,细菌质粒有着重要的生态学意义。     

人工湿地微生物生态学研究

本文从微生物生态学的角度阐述了脱氮的细菌和真菌、硫酸盐还原细菌以及和产甲烷细菌等主要功能微生物的相关研究进展,并总结了水生植物、营养物质以及基质、温度等对人工湿地中微生物的空间分布的影响,对人工湿地微生物生态学的前景进行了展望,为相关研究提供资料和借鉴。     

细菌总数测定在水生态环境中的应用探讨

本文中细菌总数指1mL水样在营养琼脂中,36±2°c培养24h后,所生长细菌菌落的总数。细菌在自然界的分布很广,广泛存在于土壤、水、空气和动植物体表面及消化道等处。因为细菌对营养物质的消耗,环境中细菌总数的多少除了反映水体中微生物污染外,同时又能指示出水体营养物质的存在情况,就细菌总数对水体水生态环境中营养物质的指示作用进行应用探讨。     

细菌气溶胶在大气冰核核化过程中作用的研究进展

介绍了大气中细菌气溶胶的种类和浓度分布的变化特征,对冰核细菌促进冰核核化的分子生物学机制,影响成冰活性的因素以及冰核在大气冰核核化过程中作用的研究进展等做了重点介绍,对冰核细菌的生物学、分子生物学和形态学及其实际应用等方面的研究进展也做了讨论,同时也指出了今后需要进一步研究的问题,尤其是生物冰核在云和降水中作用方面的问题.     

荧光素酶基因(Lux)在环境监测中的应用和研究

通过基因工程技术从发光细菌中无性繁殖荧光素酶基因(Lux),并转移到对污染物有特别敏感性的受体菌中表达,或组入对特定细菌种群有专一感染性的噬菌体基因组中,构建成基因工程菌或工程噬菌体用于环境监测,将能极大地提高监测的效率和灵敏性。该技术自80年代创立至今,已成功地用于多种污染物或细菌的监测,显示了较传统发光细菌监测法的明显优势,迅速成为污染生物监测研究的一个热点。本文综述了近年有关荧光素酶基因(Lux)的分子生物学特性及其监测应用的最新研究进展和尚存的问题。      

应用FISH法对污水生物处理系统中细菌数量的计数

近年来,FISH(Fluorescence in Situ Hybridization,荧光原位杂交)法作为一种应用分子生物学技术对细胞及细菌等微生物进行定性以及定量分析的研究方法,在国际上得到广泛的应用。本文介绍了应用FISH法对污水处理工艺中细菌数量的计数方法。同时介绍了活性污泥试样制作、FISH法观测试样的制作以及各种试剂的配制、作用以及使用方法,基因探针的选择原则、杂交方法等实际操作方法与过程。     

厌氧水环境中噬菌体MS2的存活和团聚特性及理化因素的影响

为探究病毒在厌氧水环境中的存活特性,以噬菌体MS2为模式病毒,采用双层平板法进行噬菌体MS2的定量检测,研究温度、pH值、悬浮颗粒物、乙酸等理化条件对噬菌体MS2的影响,分析其衰减动力学特征,并通过测定Zeta电位和噬菌斑直径考察噬菌体颗粒在不同条件下的聚集状况.结果表明,在厌氧水环境中,噬菌体MS2的衰减符合一阶指数衰减模型.在众多研究因素中,温度是影响噬菌体MS2存活的最主要因素.噬菌体MS2在4,17,25和35℃时的T₉₀分别为20.36,6.14,5.15和0.46d.35℃条件下12h后噬菌体失活率高达2.44lg,而4℃条件下7d后噬菌体失活率仅有0.78lg.增加乙酸浓度能明显提高噬菌体MS2的衰减速率.低pH值和悬浮颗粒物条件会促进噬菌体的团聚,使噬菌体颗粒Zeta电位降低,水力学直径增大,但悬浮颗粒物浓度过高会影响颗粒间的静电作用.噬菌体的团聚也增加了噬菌斑的直径,pH=6和20mg/L的悬浮颗粒物条件下,直径1.0mm以上的大噬菌斑数量占比分别达到了45.61%和57.74%.明确厌氧水环境中各种理化因素对噬菌体MS2的影响,可为水环境病毒控制提供科学依据.     

热泉微生物席及其成员光合细菌中的可抽提类脂物的比较分析

为了研究重要的生物标志即类脂物的分布是否与不同种类的光合细菌的分布一致,对热泉微生物席和细菌的可抽提烃类、蜡酯、醇和脂肪酸进行了对比。研究了两种蓝细菌席、两种厌氧光合细菌席及它们的成员光合细菌(Chloroflexus aurantiacus,Chromatium tepidum 和 Chlorobium sp.)。总的说来,菌席类脂反映了当时主要建席光养型微生物的输入。蓝细菌席主要含有烃类,其中以正十七烷和中部支链烷烃为主;而光合细菌席中则缺乏中部支链烷烃。除了C.aurantiacus以具有C_(29)-C_(32)不饱和的一、二和三烯烃为特征以外,光合细菌和菌席含烃不丰富。蜡酯是C.aurantiacus和Chlorobium sp.以及含这些微生物的菌席的主要成分。占优势的游离醇反映了这些菌席的光合微生物中叶绿素 a 或细菌叶绿素的分布。蓝细菌席和厌氧光合细菌席中的痕量甾醇可能是外来的,在所有的菌席中游离的脂肪酸为次要成分,且分布相似。     

生活垃圾填埋场细菌气溶胶粒径分布及种群特征

卫生填埋是一种常用的生活垃圾处置方法.在倾倒、堆放、推平和压实等垃圾填埋过程中,有大量带有致病菌的微生物气溶胶逸散,污染空气,危害人体健康.本研究在华北地区某生活垃圾卫生填埋场设置采样点,采集空气中的细菌气溶胶,解析细菌气溶胶的浓度、粒径分布和种群特征,研究空气温度、相对湿度以及风速对细菌气溶胶逸散的影响.结果表明,作业区和覆盖区空气细菌浓度分别为(5 437±572) CFU·m~(-3)和(2 707±396) CFU·m~(-3).垃圾渗滤液处理区空气中的细菌气溶胶浓度最高,平均为9 460 CFU·m~(-3).细菌气溶胶的浓度呈现明显的季节变化,夏季浓度明显高于其他季节.冗余分析(RDA)显示,气象参数如相对湿度、温度和风速,显著影响细菌气溶胶在空气中的数量.作业区和覆盖区空气细菌粒径分布高峰分别在2. 1～4. 7μm和0. 65～2. 1μm.渗滤液处理逸散的细菌气溶胶大部分大于4. 7μm. Moraxellaceae,Bacillus aerius,Arcobacter以及Aeromonas是垃圾填埋场细菌气溶胶中检出潜在或机会致病菌.     

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.     

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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Single and joint effects of pesticides and mercury on soil urease

The influence of two pesticides including chlorimuron-ethyl and furadan and mercury （Hg） on urease activity in 4 soils （meadow burozem and phaeozem） was investigated. The soils were exposed to various concentrations of the two pesticides and Hg individually and simultaneously. Results showed that there was a close relationship between urease activity and organic matter content in soil. Chlorimuron-ethyl and furadan could both activate urease in the 4 soils. The maximum increment of urease activity by chlorimuronethyl was up to 14%-18%. There was almost an equal increase （up to 13%-21%） in the urease activity by furadan. On the contrary, Hg markedly inhibited soil urease activity. A logarithmic equation was used to describe the relationship （P〈0.05） between the concentration of Hg and the activity of soil urease in the 4 tested soils. Semi-effect dose （ED50） values by the stress of Hg based on the inhibition of soil urease in the 4 soils were 88, 5.5, 24 and 20 mg/kg, respectively, according to the calculation of the corresponding equations. The interactive effect of chlorimuron-ethyl or furadan with metal Hg on soil urease was mainly synergic at the highest tested concentrations.     

Isolation and preliminary characterization of a 3-chlorobenzoate degrading bacteria

A study was conducted to compare the diversity of 2-, 3-, and 4-chlorobenzoate degraders in two pristine soils and one contaminated sewage sludge. These samples contained strikingly different populations of mono-chlorobenzoate degraders. Although fewer cultures were isolated in the uncontaminated soils than contaminated one, the ability of microbial populations to mineralize chlorobenzoate was widespread. The 3- and 4-chlorobenzoate degraders were more diverse than the 2-chlorobenzoate degraders. One of the strains isolated from the sewage sludge was obtained. Based on its phenotype, chemotaxonomic properties and 16S rRNA gene, the organism S-7 was classified as Rhodococcus erythropolis. The strain can grow at temperature from 4 to 37℃. It can utilize several （halo）aromatic compounds. Moreover, strain S-7 can grow and use 3-chlorobenzoate as sole carbon source in a temperatures range of 10-30℃ with stoichiometric release of chloride ions. The psychrotolerant ability was significant for bioremediation in low temperature regions. Catechol and chlorocatechol 1,2-dioxygenase activities were present in cell free extracts of the strain, but no （chloro）catechol 2,3- dioxygenase activities was detected. Spectral conversion assays with extracts from R. erythropolis S-7 showed accumulation of a compound with a similar UV spectrum as chloro-cis,cis-muconate from 3-chlorobenzoate. On the basis of these results, we proposed that S-7 degraded 3-chlorobenzoate through the modified ortho-cleave pathway.     

Organochlorine pesticides in soils under different land usage in the Taihu Lake region, China

A field study was conducted in the Taihu Lake region, China in 2004 to reveal the organochlorine pesticide concentrations in soils after the ban of these substances in the year 1983. Thirteen organochlorine pesticides (OCPs) were analyzed in soils from paddy field, tree land and fallow land. Total organochlorine pesticide residues were higher in agricultural soils than in uncultivated fallow land soils. Among all the pesticides, ΣDDX (DDD, DDE and DDT) had the highest concentration for all the soil samples, ranging from 3.10 ng/g to 166.55 ng/g with a mean value of 57.04 ng/g and followed by ΣHCH, ranging from 0.73 ng/g to 60.97 ng/g with a mean value of 24.06 ng/g. Dieldrin, endrin, HCB and α-endosulfan were also found in soils with less than 15 ng/g. Ratios of p,p'-(DDD DDE)/DDT in soils under three land usages were: paddy field ＞ tree land ＞ fallow land, indicating that land usage inlfuenced the degradation of DDT in soils. Ratios of p,p'-(DDD DDE)/DDT ＞1, showing aged residues of DDTs in soils of the Taihu Lake region. The results were discussed with data from a former study that showed very low actual concentrations of HCH and DDT in soils in the Taihu Lake region, but according to the chemical half-lives and their concentrations in soils in 1980s, the concentration of DDT in soils seemed to be underestimated. In any case our data show that the ban on the use of HCH and DDT resulted in a tremendous reduction of these pesticide residues in soils, but there are still high amounts of pesticide residues in soils, which need more remediation processes.     

Sorption of 1-naphthol by plant cuticular fractions

The contribution of aliphatic-rich plant biopolymer to sorption of hydrophobic organic compounds is significantly important because of their preservation and accumulation in the soil environment,but sorption mechanism is still not fully understood.In this study, sorption of 1-naphthol by plant cuticular fractions was examined to better understand the contributions of respective fraction.Toward this end,cuticular materials were isolated from the fruits of tomato by chemical method.The tomato cuticle sheet consisted of waxes (6.5 wt%),cuticular monomer (69.5 wt%),and polysaccharide (24.0 wt%).Isotherms of l-naphthol to the cuticular fractions were nonlinear (N value (0.82-0.90)) at the whole tested concentration ranges.The KodKow ratios for bulk cuticle (TC1),dewaxed cuticle (TC2),cutin (TC4),and desugared cuticle (TC5) were larger than unity,suggested that tomato bulk cuticle and cutin are much powerful solption medium.Sorption capability of cutin (TC4) was 2.4 times higher than the nonsaponifiable fraction (TC3).The 1-naphthol interactions with tomato cuticular materials were governed by both hydrophobic-type interactions and polar (H-bonding) interactions. Removal of the wax and polysaccharide materials from the bulk tomato cuticle caused a significant increase in the sorption ability of the cuticular material.There was a linear negative trend between K_(oc) values and the amount of polysaccharides or fraction's polarities ((N O)/C);while a linear positive relationship between K_(oc) values and the content of cutin monomer (linear R~2=0.993) was observed for present in the cuticular fractions.Predominant sorbent of the hydrophobic organic compounds (HOCs) in the plant cuticular fraction was the cutin monomer,contributing to 91.7% of the total sorption of tomato bulk cuticle.