理化因素及感染复数对噬菌体MS2活性的影响

噬菌体MS2作为潜在的水中肠道病毒指示生物,已逐渐成为回用水微生物学安全评价指标的主要研究方向之一。测定了温度、pH值、紫外线和感染复数对噬菌体MS2活性的影响。结果表明:噬菌体MS2在70℃时5 min内即会全部失活,在pH值4～10的范围内均可稳定存在,在短波紫外线照射下25 min内即可失活;噬菌体MS2感染其宿主大肠杆菌的最佳感染复数为1。     

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.     

紫外活化过硫酸钠灭活水中噬菌体MS2的特性及机制

水环境中的病毒对常见的消毒技术有较强的抵抗力.为了开发水中病毒的高效灭活技术,以噬菌体MS2为对象,研究紫外活化过硫酸钠（UV/PS）体系灭活病毒的特性和机制.采用双层平板法对噬菌体MS2进行定量检测,研究UV/PS对水样中噬菌体MS2的灭活率和动力学特征,并考察PS用量、pH值和噬菌体初始浓度等因素对灭活效果的影响.利用透射扫描电镜观察UV/PS处理前后噬菌体的形貌,利用电子顺磁共振波谱法确认反应体系中存在的自由基种类.在自由基淬灭实验的基础上,分析计算UV/PS体系中各因素对噬菌体灭活的贡献率.结果表明,当紫外辐照强度为160 μW·cm^-2时,UV/PS处理4 min即可去除4.39 lg的噬菌体MS2,较单独使用同样辐照剂量的UV消毒灭活率高1.44 lg.UV/PS体系对噬菌体MS2的灭活动力学过程符合一级反应动力学模型.增加体系中的PS初始浓度能明显提高对噬菌体的灭活率和灭活速率,而pH和噬菌体初始浓度对UV/PS灭活噬菌体的影响较小.UV/PS处理可导致噬菌体的衣壳破损,促进了噬菌体颗粒团聚.UV/PS体系中存在SO₄^-·和·OH,是噬菌体MS2灭活的重要因素.·OH比SO₄^-·对噬菌体MS2灭活的贡献更大.     

冠状病毒IBV和噬菌体MS2在饱和多孔介质中的运移规律

为研究病毒在土壤中的运移机理,选择鸡的传染性支气管炎冠状病毒(IBV)和噬菌体(MS2)为材料,采用土柱模拟实验研究2种病毒在不同输入方式(IBV脉冲,MS2阶跃)、不同饱和多孔介质中的运移规律.结果表明,IBV在饱和玻璃珠中的运移有少量滞留和延迟;MS2在饱和砂土中的运移随着离子强度的不同而不同,存在两个临界点,分别为0.065mol/L和0.080mol/L.在离子强度从0.002mol/L升高到0.065mol/L时,病毒吸附随着离子强度的增加而迅速增加;离子强度在0.065～0.080mol/L时,病毒吸附随着离子强度的增加而迅速降低;当离子强度超过临界点0.08mol/L后,随着离子强度的增加病毒吸附缓慢增加.在相同pH值和离子强度的不同介质中,IBV比MS2吸附和滞留较多.     

pH值对白腐真菌液体培养基抑制杂菌效果的影响研究

应用摇瓶试验研究了不同初始pH值对白腐真菌Phanerochaete chrysosporium液体培养基在非灭菌环境抑制杂菌效果的影响.结果表明,当采用Phanerochaete chrysosporium孢子作为种子在非灭菌环境进行培养时,初始pH值为3.6和4.4的液体培养基在培养第1d仅感染酵母菌,而初始pH值为5.6的液体培养基不仅感染了酵母菌而且还感染了细菌;正是由于非灭菌环境培养体系感染杂菌,使得后续Phanerochaete chrysosporium对活性艳红K-2BP的脱色能力大大降低甚至丧失;而采用灭菌环境培养Phanerochaete chrysosporium,非灭菌环境脱色活性艳红K-2BP的方法却获得了较好的脱色效果,3种初始pH值的氮限制液体培养基培养出的Phanerochaete chrysosporium对活性艳红K-2BP 45h的脱色率均在70%以上,接近或超过灭菌环境的结果,其中初始pH值为4.4的液体培养基培养的Phanerochaete chrysosporium在非灭菌环境对活性艳红K-2BP的脱色效果最好,其24h的脱色率达到80%以上.尽管3种pH液体培养基在脱色过程中也同样感染了杂菌,但与非灭菌环境培养体系相比含量很少,没有影响脱色效果.因此,可以得出低pH值(pH=3.6,pH=4.4)氮限制培养基虽然在一定程度上可以抑制细菌,但是却不能抑制酵母菌;当在非灭菌环境使用Phanerochaete chrysosporium脱色活性染料时,Phanerochaete chrysosporium只有在灭菌环境培养至菌丝体形成并在整个系统占优势,才能获得较高的脱色效果.     

藻细胞和高岭土的存在对病毒MS2存活的影响

选用病毒MS2作为水中肠道病毒的指示病毒,高岭土和铜绿微囊藻分别作为无机颗粒物和有机颗粒物,研究颗粒物浓度、pH值、不同价态离子浓度、天然有机物(NOM)等水质条件下,无机(高岭土)、有机(铜绿微囊藻)颗粒物存在对病毒MS2存活的影响.结果表明,无机颗粒物高岭土对病毒MS2的存活无明显影响,但当水体钙硬度(钙离子产生的硬度)较大时,病毒MS2的表观存活量增加1个对数;铜绿微囊藻的存在会导致病毒MS2的存活量降低1个对数左右,但当溶液的pH值大于4.0或铜绿微囊藻的浓度小于1.0×106cells·L-1时,藻类对病毒的生存无明显影响;当水体钙硬度较大时,藻反而会增加病毒MS2的存活对数.因此在高浊水、高藻水中,水的钙硬度增加会使水体中病毒生存能力变强,进而增加饮用水的安全风险.     

纳米TiO2-光催化灭活水中噬菌体MS2

提出了一种简易的水中病毒灭活方法,以噬菌体MS2为模式病毒,探讨了紫外光照射时间、TiO2浓度、紫外光强度对自来水中MS2灭活效率的影响;评价了太阳光代替紫外光用于光催化灭活病毒的可行性.研究结果表明,聚乙烯(PE)材质的样品袋能更好地透过紫外光;紫外光照射6h,50mg/L的TiO2可以灭活自来水中7.95Log的MS2;紫外光强度在45μW/m2以下时,灭活率随强度的提高而增加;在溪水中,50mg/L的TiO2仅有4.87Log的灭活效率. 在太阳光下,50mg/L的TiO2在自来水和溪水中对MS2的灭活效率差异不明显,都达到了6.1Log以上,表明太阳光-TiO2灭活水中的病毒可行且高效.     

超声、紫外增强H2O2/KI降解磺胺甲基嘧啶

实验以H_2O_2和KI作为分子碘(I2)的来源,研究比较了超声、紫外分别增强H_2O_2/KI降解磺胺甲基嘧啶(sulfamerazine,SMR)的效果、主要影响因素、分子碘的生成、主要活性物种和产物.结果表明,超声、紫外均能有效增强H_2O_2/KI对磺胺甲基嘧啶的降解,超声的增强效果明显;pH值对磺胺甲基嘧啶的去除率影响较大,pH为2.6~5.6,去除率随pH升高而降低;自由基抑制结果表明,碘自由基(I·和I-2·)是超声、紫外增强H_2O_2/KI降解磺胺甲基嘧啶的主要活性物质.HPLC/MS/MS分析检测到一碘代苯.     

纳米TiO_2-光催化灭活水中噬菌体MS

提出了一种简易的水中病毒灭活方法,以噬菌体MS2为模式病毒,探讨了紫外光照射时间、TiO2浓度、紫外光强度对自来水中MS2灭活效率的影响;评价了太阳光代替紫外光用于光催化灭活病毒的可行性.研究结果表明,聚乙烯(PE)材质的样品袋能更好地透过紫外光;紫外光照射6h,50mg/L的TiO2可以灭活自来水中7.95Log的MS2;紫外光强度在45μW/m2以下时,灭活率随强度的提高而增加;在溪水中,50mg/L的TiO2仅有4.87Log的灭活效率.在太阳光下,50mg/L的TiO2在自来水和溪水中对MS2的灭活效率差异不明显,都达到了6.1Log以上,表明太阳光-TiO2灭活水中的病毒可行且高效.     

不同分散条件和pH值对TiO_2悬浊液性能影响研究

TiO2悬浊液的稳定性对于光催化应用而言是非常重要的。对于不同粒径的纳米TiO2颗粒在超声分散和搅拌分散、不同pH条件下的团聚和沉降进行了考察,结果显示颗粒的团聚和沉降与分散形式、颗粒粒径、pH值密切相关。颗粒采用超声分散的稳定性较好,但仍会发生沉降,而颗粒粒径越大,团聚和沉降越明显;pH值的影响与距离等电点远近有关系,越近则越容易团聚和沉降,但酸性条件下的分散性要好于碱性条件。结合相关理论,对于其中的机理也进行了探讨。     

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.     

Effects of two sludge application on fractionation and phytotoxicity of zinc and copper in the soil

The potential harm of heavy metals is a primary concern in application of sludge to the agricultural land. A pot experiment was conducted to evaluate the effect of two sludges on fractionation of Zn and Cu in soil and their phytotoxicity to pakchoi. The loamy soil was mixed with 0%, 20%, 40%, 60% and 80% (by weight) of digested sewage sludge (SS) and composted sludge (SC). The additions of both sludges caused a significant raise in all fractions, resulting in that exchangeable (EXCH) and organic bound (OM) became predominance of Zn and organic bound Cu occupied the largest portion. There was more available amount of Zn and Cu in SS treatments than SC treatments. During the pot experiment, the concentration of Zn in EXCH, carbonate (CAR) and OM and Cu in EXCH and OM fractions decreased in all treatments, so their bioavailability reduced. Germination rate and plant biomass decreased when the addition rate was high and the best yield appeared in 20% mixtures at the harvest of pakchoi. The two sludges increased tissue contents of Zn and Cu especially in the SS treatments. Zn in pakchoi was not only in relationship to ΔEXCH and ΔCAR forms but also in ΔOM forms in the sludge-soil mixtures. Tissue content of Cu in pakchoi grown on SC-soils could not be predicted by ΔEXCH. These correlation rates between Zn and Cu accumulation in pakchoi and variation of different fractions increased with time, which might indicate that sludges represented stronger impacts on the plant in long-term land application.     

Intraspecific differences in effects of co-contamination of cadmium and

A hydroponic experiment was carried out to study intraspecific differences in the effects of different concentrations of cadmium (Cd)(0-10 mg/L) and arsenate (As(V)) (0-8 mg/L) on the growth parameters and accumulation of Cd and As in six wheat varieties Jing-9428, Duokang-1, Jingdong-11, Jing-411, Jingdong-8 and Zhongmai-8. The endpoints of wheat seedlings, including seed germination,biomass, root length and shoot height, decreased with increasing the Cd and As concentrations. Significant differences in seed germination, biomass, root length, shoot height and the accumulation of Cd and As were observed between the treatments and among the varieties (p ＜ 0.05). The lethal dosage 50% were about 20, 80, 60, 60, 80 and 20 mg As/L for Jing-9428, Duokang-1, Jingdong-11,Jing-411, Jingdong-8 and Zhongmai-8, respectively, and the corresponding values for Cd were about 30, 80, 20, 40, 60 and 10 mg Cd/L, respectively. Among the six varieties, Duokang-1 was found to be the most resistant to Cd and As toxicity, and Zhongmai-8 was the most sensitive to Cd and As co-contamination. The resistance of the six varieties was found dependant on the seedling uptake of Cd and As. Duokang-1 was the most suitable for cultivation in Cd and As co-contaminated soils.     

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

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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.     

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.