一株嗜油不动杆菌(Acinetobacter oleivorans)的分离鉴定及石油降解特性

从石油污染土壤分离到30株石油降解菌,经复筛得到一株适应能力强、对石油降解效率高的优势菌株JC3-1。该菌株在温度为20~40℃、p H 5~9、盐度(Na Cl)为0~3%(W/V)条件下生长良好。在以4 g/L原油为惟一碳源的培养基中生长15 d后对原油的降解率达42.15%。通过形态学观察、生理生化实验及16S r DNA序列分析,初步鉴定属于嗜油不动杆菌(Acinetobacter oleivorans)。菌株在以正戊烷、正己烷、十六烷、十八烷、苯、甲苯、二甲苯、邻苯二酚、萘、芘为惟一碳源培养基中都能很好生长。用特异性PCR检测发现,JC3-1具有酰基辅酶A脱氢酶、苹果酸合成酶、异柠檬酸裂合酶、Ton B-依赖性铁载体受体、铁载体受体蛋白、多功能脂肪酸氧化酶复合体α亚单位等代谢功能基因,及烷烃单加氧酶、芳烃双加氧酶、联苯双加氧酶、邻苯二酚双加氧酶、萘双加氧酶、甲苯双加氧酶等降解功能基因。     

一株芘降解菌B2的降解条件优化及降解基因

芳香烃降解菌是石油污染土壤修复的主要生物资源。采用芘平板升华法对克拉玛依原油污染土壤样品进行驯化培养,分离得到一株芘降解菌B2,经16S rDNA基因序列比对及系统发育进化分析表明,该菌株为假单胞菌属(Pseudo-monas)。采用正交设计方法优化菌株B2对高分子量多环芳烃芘的降解条件,并构建多元非线性模型预测菌株B2对芘的最佳降解条件,结果表明:在接种量OD660 nm为0.60、降解温度为40℃、降解时间为6.0 d时,预测菌株B2对芘的降解最大达到38.214 mg/L,实际测得最大降解量为37.906 mg/L,预测准确率为99.19%。运用PCR技术克隆B2的邻苯二酚-2,3-双加氧酶基因(B2C23O)(I.2.A亚家族),核酸序列分析表明,该基因全长880 bp,具有一个完整的开放阅读框,编码246个氨基酸,与已报道的Pseudomonas putida W619同源性最高为97%;对B2C23O基因编码氨基酸序列进行分析,发现其具有邻位断裂双加氧酶模式结构,推测菌株B2通过邻位裂解途径降解芘代谢中间产物邻苯二酚。     

海洋芘降解菌Alteromonas sp.P7的降解特性及基因组初步分析

以芘(PYR)为唯一碳源,在青岛受石油烃污染的码头的海底沉积物中分离到1株PYR海洋降解菌株P7,经过形态学观察、16S rRNA基因序列分析鉴定菌株P7属于交替单胞菌(Alteromonas)。该菌株在20 d内对PYR的降解率可达44.8%。经GC-MS鉴定分析发现,菌株P7降解芘的代谢产物中含有邻苯二甲酸和水杨酸,且细胞代谢过程中产生邻苯二酚2,3-双加氧酶,由此推测出Alteromonas sp. P7对芘的降解中间产物中含有邻苯二酚,且对芘的降解是通过邻苯二甲酸途径和水杨酸途径实现的。通过对分离得到的Alteromonas sp. P7进行测序,得到1条全长4 597 467 bp的基因组,对该基因序列注释,得到编码基因8 164个,平均GC含量为44.26%。重将测序结果与数据库比对并检索,菌株P7含有参与编码邻苯二酚2,3-双加氧酶的基因。     

一株苯胺降解菌的分离鉴定及其降解特性

为研究苯胺污染的生物控制,通过驯化培养,从南京化工厂污水处理厂的活性污泥中分离出一株高效苯胺降解菌——菌株AN4.生理生化试验鉴定菌株AN4为金黄杆菌属(Chryseobacterium sp.).菌株AN4利用苯胺生长和降解苯胺的最适温度为30 ℃、pH为7.0,它可在苯胺质量浓度低于3 000 mg/L的无机盐固体培养基上生长.菌株AN4除可降解苯胺外,还可以苯酚、苯甲酸、硝基苯、甲苯、萘、氯苯、二甲苯作为唯一碳源生长,蛋白胨可加速其对苯胺的代谢.代谢机制研究证实,菌株AN4在邻苯二酚1,2-双加氧酶作用下经邻位裂解途径降解苯胺.     

低温硝基苯降解菌选育及在土壤修复中的应用

从硝基苯污染地下水中筛选出一株以硝基苯为唯一碳源和氮源的低温、高效降解菌,命名为XJ菌;初步鉴定XJ菌为革兰氏阴性微小短杆细菌、恶臭假单胞菌属,降解硝基苯遵循部分还原降解途径;在10℃下培养96 h,XJ菌对硝基苯的降解去除率达到91.1%,其中约有67.1%的硝基氮素转变成了氨氮。在10℃下利用XJ菌修复硝基苯污染土壤,修复时间为96 h时,硝基苯去除率达到85.7%,XJ菌密度及微生物脱氢酶活性保持较高的水平。本研究可为低温下硝基苯污染土壤的生物修复提供参考。     

甲氰菊酯降解菌鉴定及其降解特性

从农药厂污泥中分离到一株能降解甲氰菊酯的光合细菌PSB07-19.鉴定结果表明,PSB07-19为沼泽红假单胞菌(Rhodopseudomonas palustris),其最适生长温度为30℃,最适pH为6.5.实验结果表明,该菌以共代谢方式降解甲氰菊酯,对甲氰菊酯的最大耐受质量浓度为600 mg/L,培养15 d对400 mg/L甲氰菊酯的降解率达45.51%,降解最适pH、温度分别为7.5、30℃.     

邻苯二酚-2,3-双加氧酶基因在芘降解菌基因组的整合与表达

为了构建能够稳定遗传且高效降解多环芳烃的工程菌,利用PCR技术对Pseudomonas songnenensis wp3-1的邻苯二酚-2, 3-双加氧酶(C23O)基因进行克隆,并将其与自杀性载体pUTmini-Tn5连接,得到重组载体pUTmini-Tn5-C23O。在三亲接合作用下,经mini-Tn5转座子将重组载体pUTmini-Tn5-C23O中的C23O基因整合到菌株Pseudomonas sp. wp4的染色体DNA中,最终得到基因工程菌wp4-C23O。在不同pH、温度下,菌株wp4和工程菌wp4-C23O对浓度为50 mg?L-1的芘进行降解7 d。2株菌降解最适温度为37℃、最适pH为7.5。在此条件下,工程菌wp4-C23O对芘降解率显著高于wp4菌株(P0.05),降解率提高11.45%。以PAHs降解优势菌株为受体构建工程菌可以去除石油污染土壤中的PAHs。     

一株耐盐N,N-二甲基甲酰胺降解菌的分离及其降解特性研究

以N,N-二甲基甲酰胺(DMF)作为唯一碳、氮源,从高盐环境中筛选耐盐DMF高效降解菌DMF-4,并对其降解DMF的特性进行研究。经鉴定,DMF-4为环太芽孢杆菌(Bacillus circulans),其在盐度为3%,温度为30℃的条件下培养48 h,对1 000 mg/L DMF的降解率可达47.7%。DMF-4降解DMF的适宜条件为初始pH 6.0～7.0,DMF初始质量浓度3 000～4 000 mg/L,温度30～40℃,盐度1%～3%;葡萄糖和醋酸钠可明显促进菌株DMF-4的生长和对DMF的降解效果。菌株DMF-4对高盐与高温皆有较好的耐受性,在工业污水处理领域具有广阔的应用空间。     

稠油降解微生物的筛选鉴定及其降解特性

从二连油田原油和油层水中筛选驯化出3株能够降解稠油的细菌DS1、DS2和DS3,通过16S rRNA基因序列比对发现DS1、DS2和DS3分别与溶血不动杆菌(Acinetobacter haemolyticus)、鹑鸡肠球菌(Enterococcus gallinarum)和耳炎短杆菌(Brevibacterium otitidis)相似度最高,分别为99%、99%和98%。研究结果表明,DS1对温度和pH有较强耐受性,DS3对盐度的适应性较好,2株菌最适的降解条件为温度35~40℃、盐度2%~5%(W/V)、pH为7~10。在5%的原油浓度下,复合菌对原油的30 d降解率达89.2%。经GC-MS分析,微生物降解作用后,除C29其他烃类几乎被全部降解。3株菌在7d内对500 g/L粘度为1 746 mPa·s(50℃)的稠油降粘率分别为49.1%、46.6%和49.0%,而复合菌对稠油的降粘效果高于单一菌株,其降粘率达到57.0%。     

华北某油田石油降解菌的筛选及降解特性

以石油为唯一碳源,从华北某油田污染土壤中筛选出石油降解菌12株。其中5株菌有较强降解能力,分别编号为z-3、z-6、z-7、z-8和z-b,在30℃,160 r/min摇床培养10 d后,菌株对石油降解率分别为63.8%、34.2%、44.8%、50.5%和42.3%。通过生理生化和分子生物学鉴定,确定这5株菌分别属于假单胞菌属(Pseudomonas)、根瘤菌属(Rhizobium)、假黄单胞菌属(Pseudoxanthomonas)、假单胞菌属(Pseudomonas)、鞘氨醇单胞菌属(Sphingomonas)。通过对这5株菌生物量、脱氢酶及邻苯二酚2,3-双加氧酶活性的监测,发现生物量及酶活性与石油降解能力具有直接关系。最后,对5株菌的生长条件进行优化,其中z-3、z-6、z-7和z-b菌株对石油降解率较优化前提高10%左右。     

Degradation of diuron in aqueous solution by ozonation

Degradation of diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea] in aqueous solution and the proposed degradation mechanism of diuron by ozonation were investigated. The factors that affect the degradation efficiency of diuron were examined. The generated inorganic ions and organic acids during the ozonation process were detected. Total organic carbon removal rate and the amount of the released Cl^? increased with increasing ozonation time, but only 80.0% of the maximum theoretical concentration of Cl^? at total mineralization was detected when initial diuron concentration was 13.8 mg L^?1. For N species, the final concentrations of NO₃ ^? and NH₄ ⁺ after 60 min of reaction time were 0.28 and 0.19 mg L^?1, respectively. The generated acetic acid, formic acid and oxalic acid were detected during the reaction process. The main degradation pathway of diuron by ozonation involved a series of dechlorination-hydroxylation, dealkylation and oxidative opening of the aromatic ring processes, leading to small organic species and inorganic species. The degradation efficiency of diuron increased with decreasing initial diuron concentration. Higher pH value, more ozone dosage, additive Na₂CO₃, additive NaHCO₃ and additive H₂O₂ were all advantageous to improve the degradation efficiency of diuron.     

1，4-二氯苯降解菌的分离及其降解特性研究

从某污水处理曝气池的活性污泥中分离出一株能够以1,4-二氯苯为唯一碳源和能源生长的菌株DEB-1,通过形态特征和生理生化试验初步鉴定为黄杆菌属（Flavobacterium sp.）。实验结果表明,该菌株最适降解温度为32℃、最适降解pH为7.8,24 h对100 mg/L的1,4-二氯苯的降解率达94.5%。菌株DEB-1的降解谱较广,对5种氯苯类物质具有较高的降解率。并进一步研究了DEB-1的1,4-二氯苯降解酶粗酶液的性质,其最适反应温度和pH分别为30℃和8.5。对处理含氯代芳香化合物的有机废水具有一定的意义。     

甲醛降解菌的筛选及降解特性研究

从采集活性污泥中筛选得到1株具有高效降解甲醛能力的菌株并命名为JQ-1,根据其形态特征,初步判断菌株JQ-1属假单胞菌属。同时对菌株JQ-1的生长特性及降解特性进行了初步研究。实验结果表明,该菌株降解甲醛的最适条件为：甲醛废水浓度为50mg／L,pH值为6,培养温度为25℃,摇床转速为150r／min。在最适条件下,菌株JQ-1具有较强的降解甲醛能力,当甲醛废水浓度为50mg／L时,在24h内甲醛降解率可达87％以上。     

一株假单胞菌降解石油污染物的降解动力学与降解机制

从某石油污染场地土壤中分离出一株石油降解菌B-1(Pseudomonas),研究了该菌株降解石油污染物的动力学,通过气相色谱(GC)/质谱(MS)分析了石油组分降解前后的变化规律,并对其降解机制进行了初探。结果表明:(1)苯、二甲苯、萘的降解动力学拟合方程大部分与一级降解动力学方程拟合效果良好,苯、二甲苯、萘的降解半衰期分别为0.47~1.67、1.54~3.60、4.41~7.00d;(2)可以通过全扫描图谱及检索出的代谢产物明确微生物降解苯、二甲苯、萘的降解途径。     

含氯苯废水的超声降解研究

以氯苯模拟废水为考察对象,研究其浓度、pH、超声时间等因素对氯苯超声降解效率的影响。实验结果表明：超声反应时间小于5min,氯苯降解率很低;5－20min,氯苯降解率成线性增大;超过20min后,氯苯降解率增大缓慢。pH值的增大,氯苯降解率略有减小。水浴温度提高,氯苯降解率增大。反应时间小于15min,初始浓度低的溶液,降解率增加较快,反应时间大于15min,初始浓度高的溶液,降解率增加较快。     

Aerobic and anaerobic degradation of profluralin and trifluralin

Abstract

The degradation of profluralin [N‐(cyclopropylmethyl)‐α,α,α‐trifluoro‐2,6‐dinitro‐N‐propyl‐]p‐toluidine] and trifluralin (α,α,α‐trifluoro‐2,6‐dinitro‐N,N‐dipropyl‐p‐toluidine) was studied under aerobic and anaerobic soil conditions. Three soils (Goldsboro loamy sand, Cecil loamy sand, Drummer clay loam) were each treated with 1 ppmw herbicide; anaerobic conditions were maintained by flooding. Soil samples were extracted monthly and subjected to TLC analysis. No degradation was detected in sterile controls. Aerobic degradation of both herbicides was greatest in the Cecil loamy sand soil over the entire incubation period. Degradation of profluralin in Cecil soil under aerobic conditions was 86 percent after 4 months with three products detected; 83 percent of the trifluralin was degraded with two products detected. Anaerobic degradation accounted for 72 percent of the profluralin and 78 percent of the trifluralin after 4 months. Degradation of both herbicides increased with incubation time for the first 3 months and decreased slightly thereafter. Generally there was more extensive degradation (percent and in number of products formed) of profluralin than trifluralin under the conditions tested. More degradation products were detected for both herbicides under aerobic conditions than under anaerobic conditions.     

一株木材腐朽菌及其粗酶液对七氯的降解

Phlebia acanthocystis TMIC34875是一株具有七氯降解能力的木材腐朽菌。为利用微生物技术去除环境中的七氯残留提供理论依据,研究了该菌株及其粗酶液对七氯的降解性能及其动力学特性。结果表明,菌株在七氯的初始浓度为50μmol/L时具有最大降解速率,为0.3031μmol/（L·h）;而菌体接种量为15%时,降解速率达到最高,为0.2045μmol/（L·h）。降解酶定位研究表明,七氯的降解主要是胞内酶在起作用。七氯胞内酶降解的酶促反应最适温度是35℃,在30-40℃之间有较高的催化活性;最适pH值为5.0,在pH 4.5-6.0之间有较高的催化活性,最适条件下反应1 h后七氯的降解率为65%。胞内粗酶液降解七氯的米氏常数K m为5.42μmol/L,最大反应速率V max为4.55μmol/min。胞内酶处理体系的GC/MS图谱显示,主要降解产物为1-羟基六氯、1-羟基-2,3-环氧六氯和环氧七氯,表明胞内酶对七氯的初始代谢机理同菌株相似,均是通过环氧化和置换反应来完成的。     

Persistence and biodegradation of carbaryl in soils

Abstract

The persistence of the methylcarbamate pesticide carbaryl was studied in four soils under flooded conditions. A substantial portion of the pesticide was recovered from all soils even after 15 days of its application, with the recovery ranging from 37% in an alluvial soil to 73% in an acid sulfate soil. The degradation of carbaryl was more rapid under flooded conditions than under nonflooded conditions. A bacterium, Pseudomonas cepacia, isolated from a flooded soil amended with a related methylcarbamate pesticide carbofuran, degraded carbaryl in a mineral medium supplemented with yeast extract.     

Fate of CL-20 in sandy soils: degradation products as potential markers of natural attenuation

Hexanitrohexaazaisowurtzitane (CL-20) is an emerging explosive that may replace the currently used explosives such as RDX and HMX, but little is known about its fate in soil. The present study was conducted to determine degradation products of CL-20 in two sandy soils under abiotic and biotic anaerobic conditions. Biotic degradation was prevalent in the slightly acidic VT soil, which contained a greater organic C content, while the slightly alkaline SAC soil favored hydrolysis. CL-20 degradation was accompanied by the formation of formate, glyoxal, nitrite, ammonium, and nitrous oxide. Biotic degradation of CL-20 occurred through the formation of its denitrohydrogenated derivative (m/z 393 Da) while hydrolysis occurred through the formation of a ring cleavage product (m/z 156 Da) that was tentatively identified as CH₂N-C(N-NO₂)-CHN-CHO or its isomer N(NO₂)CH-CHN-CO-CHNH. Due to their chemical specificity, these two intermediates may be considered as markers of in situ attenuation of CL-20 in soil.     

Molarity provides better clarity – using molar-based data when evaluating chlorinated volatile organic compound impacted sites

Abstract

Chlorinated volatile organic compounds are common constituents observed at many contaminated groundwater sites. Common industry practice has been to measure these constituents in groundwater on a weight concentration basis (e.g. µg/l, mg/l). This paper highlights the use of molar-based concentrations, especially in the case of parent-daughter degradation sequences commonly observed with the chloroethene, chloroethane, and chloromethane families of compounds. Converting to molar-based concentrations provides the practitioner greater insight into groundwater plume behavior including better evaluation of degradation processes, remedial progress, possible commingling, and/or sourcing. For example, this paper provides a tank analogy to evaluate whether the commonly misinterpreted observation of “DCE-stall” may be occurring at a site. Multiple examples of the benefits of using molar-based concentration data are also summarized in a project case study presented herein. As demonstrated in this paper, molarity does provide better clarity and can be a powerful evaluation tool in the groundwater practitioner’s toolbox.