蛋白酶对水环境中病毒的灭活作用

以大肠杆菌噬菌体T₄作为模式病毒,研究了蛋白酶对病毒的灭活作用.试验结果表明:蛋白酶灭活病毒的效果明显. 适宜条件下,67.5u/mL的蛋白酶K处理1h对纯水中病毒(6.2×10⁵PFU/L)和生活污水中病毒(7.2×10⁵PFU/L)灭活率分别达到了99.4%和49.4%,处理3h的灭活率分别是>99.9%和81.1%;工业蛋白酶1398在75.0u/mL酶浓度下处理1h对纯水水中病毒(1.7×10⁵ PFU/L)灭活率达到74.4%.pH和温度对病毒灭活效果的影响不明显.     

无排泥运行MBR处理不同浓度氨氮废水及其生物特性

考察了水力停留时间(HRT)为10 h时无排泥运行下膜-生物反应器(MBR)处理不同浓度氨氮无机废水的运行性能及微生物特性.结果表明,在处理NH₄⁺-N≤500 mg/L的废水时,氨氮转化率可达99%且反应器内微生物增长缓慢,比硝化速率从0.2kg/(kg·d)升至0.52 kg/(kg·d);当NH₄⁺-N≥700 mg/L时,出水亚硝酸氮和氨氮相继出现明显累积,比硝化速率随之下降至0.4kg/(kg·d)以下,反应器内生物量从3 200 mg/L上升至6 700 mg/L.反应器中的氨氧化菌维持在10⁷CFU/mL,亚硝酸盐氧化菌从10⁶CFU/mL下降至10³CFU/mL.系统内的溶解性微生物产物(以TOC表示)升至65 mg/L后保持稳定,出水TOC一直维持在3～4mg/L;细胞外分泌物(EPS)在膜的截流作用下随运行时间的延长积累至600 mg/L.     

三阶段控温堆肥过程中接种复合微生物菌群的变化规律研究

接种复合微生物是提高堆肥效率的主要方法之一,但由于堆料中土生微生物的竞争,较高浓度的土生微生物浓度会抑制接种微生物的长生繁殖.实验表明:当堆料中土生微生物初始浓度为4×10⁸CFU/g时,所接种的微生物不增殖,且随着堆肥的进行,浓度下降很快;而非接种微生物增殖很快,最高浓度可达10¹⁰CFU/g.当土生微生物浓度降低至4×10⁵个/g,接种微生物增殖较快,最高达10¹¹CFU/g.因此,本文利用堆肥自身产热和少许外来热源加热的三阶段控温法进行堆肥.使堆温在4 h内迅速升到70℃以上,并维持8 h,从而使土生微生物浓度降至4×10⁵个/g以下,并起到软化堆料,便于微生物降解的目的.待温度冷却至35℃～45℃时,接种复合微生物,使其快速生长繁殖,数量从10⁸ CFU/g上升到10¹¹CFU/g(干样品),且接种微生物以非接种做生物保持优势地位,从而快速分解垃圾中的有机物.由于在最终产品中含有大量接种有益微生物,可利用其作为菌肥进行回流接种堆肥,以增加堆料中接种微生物数目、改善堆肥微环境、节约菌剂用量、缩短堆肥发酵周期和降低堆肥成本.但利用接种微生物堆肥产品作为菌肥反复接种时,随着反复次数的增加,非接种微生物高浓度繁殖;接种微生物和非接种微生物浓度比约为1:3(反复接种5次),浓度情况发生了逆变.因此,回流菌肥反复接种5次后,接种菌剂基本上就不再起作用了.     

氯胺消毒对铜和不锈钢管壁生物膜的控制作用

以松花江为水源的哈尔滨某水厂出厂水为研究对象,采用生物膜培养反应器(rotating annular bioreactor, RAB)模拟给水管网系统,考察了氯胺对铜和不锈钢2种管材上生物膜的控制情况.结果表明,在无氯胺情况下运行时,铜和不锈钢挂片上的生物膜在第18d和21 d达到最大值,其生物量分别为5.5×１０³ CFU/cm²和2.5×１０⁵ CFU/cm²,生物膜达到稳定时生物量分别为1.0×１０³ CFU/cm²和1.3×１０⁵ CFU/cm²,铜挂片上的生物量明显低于不锈钢.反应器在有氯胺状态下运行时,铜和不锈钢挂片上最大生物量为5.0×10² CFU/cm²和5.0×１０⁴ CFU/cm²,分别低于无氯胺情况下1个数量级,达到稳定状态时,其生物量分别为10 CFU/cm²以下和3.5×１０⁴ CFU/cm²,说明氯胺对处于稳定状态的生物膜具有一定的控制作用,对铜挂片上的生物膜具有更明显的控制效果.对消毒剂投量、管材和管壁生物膜HPC进行分析可知,生物膜HPC与余氯胺具有良好的指数相关关系,增加氯胺投量可以降低生物膜HPC数量,氯胺和铜挂片对管壁上的生物膜具有协同灭菌作用.     

Mn2+在黄杆菌FCN2菌株降解芘过程中的作用研究

利用紫外分光光度法和原子吸收分光光度法研究了Mn²⁺在菌株FCN2生长细胞、悬浮细胞、粗酶液降解芘时的影响作用.在菌体生长期加入0.1～5 mmol/L的Mn²⁺,对菌体的生长及生长量无影响,但对芘的降解均有促进作用,以加入0.1mmol/L的Mn²⁺效果最好,其对芘的平均降解率是对照的1.26倍.此时菌株富集的Mn²⁺为0.025mmol/L;在用无外加Mn²⁺培养的菌株FCN2悬浮细胞降解芘时,加入0.5 mmol/L的Mn²⁺,芘的平均降解率为对照的1.67倍.降解反应发生72h后菌株富集的Mn²⁺为0.18mmol/L;在酶促降解时加入0.1mmol/LMn²⁺,平均降解率为对照的1.30倍.结果表明,在不同时期加入的Mn²⁺对降解芘均有一定的促进作用.     

渤海湾天津沿岸海水中甲肝病毒的检测和定量

甲肝病毒（hepatitis A virus, HAV）是能引起传染性甲型肝炎的单链RNA病毒.用常规RT-PCR（反转录PCR）和SYBR Green实时定量RT-PCR方法,根据甲肝病毒保守的VP1-VP2基因序列设计引物,对渤海湾天津沿岸海水中的甲肝病毒进行了检测和定量分析. 9个样品取自渤海湾天津塘沽以南沿岸海水,取样时间分别是2007年夏、秋、冬季和2008年春季. 海水样品先用小型超滤装置（Millipore Pellicon Mini TFF）或超滤离心管（Millipore Centricon Plus-70）浓缩,然后进行RT-PCR检测.结果表明,从9个海水样品中都能扩增出192 bp的HAV cDNA,这些cDNA的核苷酸序列与GenBank中的同源序列相似性为95%～100%. 用SYBR Green 实时定量RT-PCR检测了春季和冬季的6个海水样品,结果表明,海水中甲肝病毒的浓度范围为5.35×10⁶～4.51×10⁷ virus particles/L.     

免疫磁珠分离与实时定量PCR技术联合检测水中轮状病毒的研究

建立了一种免疫磁珠分离技术联合实时定量PCR快速定量检测水中轮状病毒的方法.通过制备能够分离水中轮状病毒的特异免疫磁珠,优化分离条件,建立了免疫磁珠分离前处理方法,并与逆转录、实时定量PCR结合,成功用于水中轮状病毒的检测.研究发现,在1 mL水样中加入10 μL轮状病毒免疫磁珠、0.25 μL Tween 20、孵育2 h可达到较好的分离效果;免疫磁珠可用于3%牛肉浸膏等常用病毒洗脱液中的轮状病毒的分离,表明该分离技术能与已有的病毒浓集方法良好地整合.免疫磁珠分离技术与实时定量PCR联合用于检测水中轮状病毒,全过程需时约5 h,检测限为1×10⁴　copies/mL(相当于3～4　PFU/mＬ),检测结果与细胞病变试验检测结果有良好的线性相关关系（R²=0.981 6）,表明其能较好地表征水样的病毒感染风险.对接种已知量的轮状病毒的污水处理厂二级出水、再生水、地表水、自来水等水样的实验表明,该法可用于各种水样中轮状病毒的检测.     

富营养化水体中浮游病毒与浮游植物生长的关系

于2005年4～12月对武汉东湖落雁岛和关桥2个池塘进行综合调查,考察浮游病毒丰度和叶绿素a含量的变化,以及水温、pH值、溶解氧量、总氮和总磷等环境因子的变化,分析浮游病毒与浮游植物、环境因子之间的相互关系.结果表明,在营养水平较高的水体中,浮游病毒丰度相应较高,但与营养物浓度变化无显著的相关性.2个采样点浮游病毒的平均丰度分别为2.99×10⁸,4.24×10⁸VLPs/mL(峰值出现在9、10月份).叶绿素a浓度的平均值分别为96.66,166.74μg/L.关桥采样点的浮游病毒丰度与前一个月的叶绿素a浓度呈显著相关(r=0.809,P<0.01),其显著性水平高于与当月叶绿素a浓度的相关性(r=0.634,P<0.05),而浮游病毒丰度与后一个月的叶绿素a浓度不呈显著相关(P>0.05).     

Cd2+ removal from wastewater by sulfate reducing bacteria with an anaerobic fluidized bed reactor

A process of treatment for containing Cd²⁺ wastewater by sulfate reducing bacteria with upflow anaerobic fluidized bed reactor has been studied. When the concentration of COD and Cd²⁺ in the influent were 270 5mg/L and 100mg/L respectively and hydraulic retention time was 4 hours, the removal rate of COD and Cd²⁺ were higher than 73 8% and 99 8% respectively. The reactor can treat as high as 1000mg/L of concentration of Cd²⁺ . The highest removal velocity rate of Cd²⁺ reached 2999 1mg/(L·d). And the possible relationship between sulfate reducing bacteria and methanogenic bacteria was discussed.     

酸性土壤环境石油烃生物降解效应

污染场地酸性土壤环境和生物修复土壤酸化,使得酸性土壤环境石油烃生物降解效应影响与有效调控成为污染土壤修复的重要内容.本文通过监测酸性和偏碱性土壤中微生物数量、活性以及石油烃含量变化,探讨酸性环境对除油微生物及烃降解效率的影响.通过投加富集菌液和生物载体,调控酸性土壤微生态环境,揭示微环境调控对于烃生物降解效应影响.研究结果表明,pH为5.4～5.7的酸性土壤,对土著除油微生物活性和数量具有显著抑制性,烃降解处于停滞状态.投加富集菌液未能有效地减弱酸性环境对除油微生物的强烈抑制作用.微生物数量在14d内从10⁶个/ g减至0,微生物FDA(FluoresceinDiacetate)活性很低,约0.10Abs/g .生物载体的投加,能有效改善介质界面微生态环境,明显减弱酸性环境的抑制效应,减缓除油微生物死亡速率.19d时土壤中降解微生物由原来的2×10⁶个/g下降到2.2×10²个/g ,第49d石油烃的生物降解率为13.02%.     

Minimizing Bias in Virally Seeded Water Treatment Studies: Evaluation of Optimal Bacteriophage and Mammalian Virus Preparation Methodologies

One key assumption impacting data quality in viral inactivation studies is that reduction estimates are not altered by the virus seeding process. However, seeding viruses often involves the inadvertent addition of co-constituents such as cell culture components or additives used during preparation steps which can impact viral reduction estimates by inducing non-representative oxidant demand in disinfection studies and fouling in membrane assessments. The objective of this study was therefore to characterize a mammalian norovirus surrogate, murine norovirus (MNV), and bacteriophage MS2 at sequential stages of viral purification and to quantify their potential contribution to artificial oxidant demand and non-representative membrane fouling. Our results demonstrate that seeding solvent extracted and 0.1 micron filtered MNV to ~10⁵ PFU/mL in an experimental water matrix will result in additional total organic carbon (TOC) and 30 min chlorine demand of 39.2 mg/L and 53.5 mg/L as Cl₂, respectively. Performing sucrose cushion purification on the MNV stock prior to seeding reduces the impacts of TOC and chlorine demand to 1.6 and 0.15 mg/L as Cl₂, respectively. The findings for MNV are likely relevant for other mammalian viruses propagated in serum-based media. Thus, advanced purification of mammalian virus stocks by sucrose cushion purification (or equivalent density-based separation approach) is warranted prior to seeding in water treatment assessments. Studies employing bacteriophage MS2 as a surrogate virus may not need virus purification, since seeding MS2 at a concentration of ~10⁶ PFU/mL will introduce only ~1 mg/L of TOC and ~1 mg/L as Cl₂ of chlorine demand to experimental water matrices.     

Surveillance of Enteric Viruses and Thermotolerant Coliforms in Surface Water and Bivalves from a Mangrove Estuary in Southeastern Brazil

This study was conducted to evaluate the microbiological quality of a mangrove estuary in the Vitória Bay region, Espírito Santo, Brazil. We analyzed the presence and concentration of enteric viruses and thermotolerant coliforms in water, mussels (Mytella charruana and Mytella guyanensis), and oysters (Crassostrea rhizophorae), collected over a 13-month period. Human adenovirus, rotavirus A (RVA), and norovirus genogroup II were analyzed by quantitative PCR. The highest viral load was found in RVA-positive samples with a concentration of 3.0 × 10⁴ genome copies (GC) L⁻¹ in water samples and 1.3 × 10⁵ GC g⁻¹ in bivalves. RVA was the most prevalent virus in all matrices. Thermotolerant coliforms were quantified as colony-forming units (CFU) by the membrane filtration method. The concentration of these bacteria in water was in accordance with the Brazilian standard for recreational waters (< 250 CFU 100 mL⁻¹) during most of the monitoring period (12 out of 13 months). However, thermotolerant coliform concentrations of 3.0, 3.1, and 2.6 log CFU 100 g⁻¹ were detected in M. charruana, M. guyanensis, and C. rhizophorae, respectively. The presence of human-specific viruses in water and bivalves reflects the strong anthropogenic impact on the mangrove and serves as an early warning of waterborne and foodborne disease outbreaks resulting from the consumption of shellfish and the practice of water recreational activities in the region.

     

三价砷对藻类群落结构的影响

为了探讨三价砷对微型生物群落结构的影响,进而评价水质,采用Ｃａｉｒｎｓ提出的ＰＦＵ方法在群落级水下上模拟研究了Ａｓ＾３＋对藻类群落的毒性,结果表明,藻类类群随着Ａｓ＾３＋浓度增大而减少,多样性指数随Ａｓ＾３＋浓度增加而明显下降,藻类群落迁入速率随时间延长而下降,消失速度则随时间而上升,Ａｓ＾３＋对藻类群落结构在ＬＯＥＣ为３２ｍｇ／Ｌ和５６ｍｇ／ＬＮＯＥＣ为１ｍｇ／Ｌ,结果表明用藻类群落在ＰＦＵ上群     

Quantitative determination of Escherichia coli based on the electrochemical measurement of bacterial catalase activity using H2O2-selective organic/inorganic-hybrid sol-gel film-modified Pt electrode

Quantitative determination of Escherichia coli (E. coli) concentration was achieved by measuring the intrinsic catalase activity of E. coli using novel H₂O₂-selective organic/inorganic-hybrid sol-gel film-modified platinum (Pt) wire electrode. This hybrid sol-gel film is composed of three kinds of organosilanes and two biopolymers (i.e., chitosan and bovine serum albumin), and exhibited an excellent permselectivity toward H₂O₂ based on a size-exclusive mechanism. The steady-state anodic current for 100 [xmol/L H₂O₂ at +0.6 V (vs. Ag/AgCl) in 0.1 mol/L phosphate buffer (pH 6.5) solution was apparently diminished by the addition of E. coli samples, due to the decomposition of H₂O₂ by intrinsic catalase activity of E. coli. The time-dependent decrease in current (–AI/At) was significantly dependent on the E. coli concentration. The –AI/At was enhanced by the permeabilization pretreatment of E. coli samples with the mixed solution of polymyxin B and lysozyme. This H₂O₂-selective organic/inorganic-hybrid sol-gel film-modified platinum (Pt) wire electrode allowed quantitative determination of E. coli concentration ranging from 10⁶ to 10⁹ CFU/mL within 30 min. This method required no label and complicated procedure, and allowed rapid, simple and cost-effective quantitative electrochemical determination of catalase-positive bacteria.     

Kinetics of phenol and m-cresol biodegradation by an indigenous mixed microbial culture isolated from a sewage treatment plant

An acclimatized mixed microbial culture, predominantly Pseudomonas sp., was enriched from a sewage treatment plant, and its potential to simultaneously degrade mixtures of phenol and m-cresol was investigated during its growth in batch shake flasks. A 2² full factorial design with the two substrates at two different levels and different initial concentration ranges (low and high), was employed to carry out the biodegradation experiments. The substrates phenol and m-cresol were completely utilized within 21 h when present at low concentrations of 100 mg/L for each, and at high concentration of 600 mg/L for each, a maximum time of 187 h was observed for their removal. The biodegradation results also showed that the presence of phenol in low concentration range (100–300 mg/L) did not inhibit m-cresol biodegradation. Whereas the presence of m-cresol inhibited phenol biodegradation by the culture. Moreover, irrespective of the concentrations used, phenol was degraded preferentially and earlier than m-cresol. A sum kinetics model was used to describe the variation in the substrate specific degradation rates, which gave a high coefficient of determination value (R² > 0.98) at the low concentration range of the substrates. From the estimated interaction parameter values obtained from this model, the inhibitory effect of phenol on m-cresol degradation by the culture was found to be more pronounced compared to that of m-cresol on phenol. This study showed a good potential of the indigenous mixed culture in degrading mixed substrate of phenolics.     

聚丙烯酰胺生物降解研究

研究了黄孢原毛平革菌对聚丙烯酰胺(PAM)的生物降解,从葡萄糖的加入量、pH、N浓度、Mn²⁺浓度和降解时间5个方面考察了对PAM降解的影响.结果表明,黄孢原毛平革菌对聚丙烯酰胺具有特殊的酶催化降解的能力,降解率可达50%,限氮条件(NH₄⁺=0.2g/L)和Mn2+浓度(Mn²⁺=0.017 5g/L)是菌株产聚丙烯酰胺降解酶的最佳条件.     

海水中原油对双齿围沙蚕(Perinereis aibuhitensis)的急性毒性效应及其体内抗氧化酶活性的影响

近年来海洋溢油事故频发,增加了溢油污染对海洋生物及海洋生态系统的暴露风险.为阐明海洋溢油事故对海洋生物的毒性效应,以双齿围沙蚕（Perinereis aibuhitensis）为受试生物,研究了不同质量浓度的海水原油溶液对双齿围沙蚕的急性毒性效应及抗氧化酶活性的影响.结果表明:双齿围沙蚕的死亡率与海水中原油暴露浓度呈正相关,暴露24、48及72 h的LC₅₀（半致死浓度）分别为13.08、9.80和8.37 g/L;暴露72 h后,双齿围沙蚕组织液中MDA（丙二醛）的含量均高于对照组,其中1.0 g/L暴露组显著高于对照组（P < 0.05）,且随暴露浓度的增大而升高;GST（谷胱甘肽巯基转移酶）的活性被显著诱导,且随暴露浓度的增大而升高;0.1 g/L原油暴露下,CAT（过氧化氢酶）和SOD（超氧化物歧化酶）的活性被显著诱导,当暴露浓度达到0.5 g/L时,CAT的诱导程度减弱,而SOD则受到抑制.研究显示,海水中的原油对双齿围沙蚕具有显著急性毒性效应,且0.1 g/L的原油暴露即可导致双齿围沙蚕产生脂质过氧化,脂质过氧化程度随暴露浓度的升高而增强;0.1 g/L原油暴露能显著诱导双齿围沙蚕体内抗氧化酶的活性,且随着海水中原油暴露浓度的增加,GST的诱导程度增强,CAT的诱导程度减弱,SOD的活性则受到抑制.      

Involvements of chloride ion in decolorization of Acid Orange 7 by activated peroxydisulfate or peroxymonosulfate oxidation

The effects of Cl^- on the decolorization of AO7 by SO₄^·- based-peroxydisulfate or peroxymonosulfate oxidation under various activated conditions were different.     

斜生栅藻对低浓度无机磷去除和生长情况的研究

研究分析了低磷浓度培养条件对斜生栅藻生长情况的影响,以及斜生栅藻对磷的去除效果.结果表明,斜生栅藻在初始细胞浓度为1×105个/mL时,可以在22h内将初始浓度0.02～0.10mg/L的磷全部去除.在初始磷浓度0.02～0.10mg/L范围内,随磷浓度的增加藻体的生长速度增加,且最大生物量也明显增大.研究发现磷浓度对斜生栅藻的形态有重要影响,在外源磷充足的条件下,斜生栅藻多为四聚体,但随着磷浓度的逐渐下降,藻细胞从四聚体,分离为二聚体,最后以单聚体为主要存在形式.     

Cadmium removal from wastewater by sponge iron sphere prepared by charcoal direct reduction

Sponge iron sphere (SIS), made of concentrated iron powder and possessed high activity and intension, was prepared through the process of palletizing, roasting and direct reduction by charcoal. The sponge iron sphere could remove most of Cd²⁺ from wastewater. The results showed the Cd²⁺ removal followed the first order reaction. Initial pH value played an important role in Cd²⁺ removal. With original initial pH, Cd²⁺ removal decreased to the minimum and then increased slightly with the rising of original concentration. The removal rate constant was ?0.1263 and ?0.0711 h^?1, respectively, under the Cd²⁺ concentration of 50 and 200 mg/L. When the initial pH was adjusted to 3.0, the removal rate constant could increase to ?9.896 and ?4.351 h^?1, respectively. The removal percentage almost reached to 100% when Cd²⁺ concentration was below 100 mg/L. While Cd²⁺ concentration was above 100 mg/L, Cd²⁺ removal percentage decreased slightly. In dynamic experiments, the column filled with sponge iron sphere exhibited favorable permeability. There was no sphere pulverization and conglutination between spheres. In contrast to the static state experiments, the Cd²⁺ removal percentage in dynamic state experiment was lower, and the removal Cd²⁺ quantity was 1.749 mg/g.