丙烯酰胺降解细菌的质粒检出及生长特性研究

从化工厂的活性污泥中筛选出具有降解丙烯酰胺能力的细菌12株,经鉴定均为不动杆菌属。碱裂解法抽提显示质粒检出率为75%,并在质粒量上有所差异。在实验室条件下,这些菌株中对丙烯酰胺的最大耐受力可达40mg/mL;12株菌株都对氨苄青霉素有抗性,而对链霉素、卡那霉素、庆大霉素及氯霉素无抗性;在以丙烯酰胺为唯一碳源的无机盐培养基中,菌株DAW01在30℃,108γ/min旋转式摇床振荡培养6d后,对丙烯酰胺降解能力可达45.01%。     

环境样品中亚硝酸细菌amoA基因的克隆与测序

对从环境样品中分离的亚硝酸细菌(Ammonia oxidizingbacteria)amoA基因进行克隆与测序,为构建基因工程菌打下基础。采用亚硝酸细菌选择性培养基,从4个不同的畜牧养殖污水处理厂采集的样品(分别编号为1,2,3,4)在室温下富集培养2个月后,采取酚氯仿抽提的方法提取DNA。根据已报道的亚硝化单胞菌(Nitrosomonassp.)amoA基因序列,设计引物AMOB AMOE,并在AMOB,AMOE的5′-端分别加上了BamHⅠ和HindⅢ的限制性酶切位点,以利于进一步酶切和克隆。用AMOB AMOE对4种样品的DNA进行PCR扩增,PCR产物进行琼脂糖凝胶电泳分析。结果表明,4种样品中1号和3号样品扩增得到预期长度的DNA片段,2号和4号样品扩增没有得到预期片段。回收纯化PCR产物与pGEM-T载体连接,构建amoA基因测序载体,并转化E.coliM15。测序结果提交GenBank进行Blast分析。结果显示,扩增得到的DNA片段均与Nitrosomonassp.GH22的amoA基因有99 7%的同源性,可从环境中分离的亚硝酸细菌中克隆出amoA基因。      

渤海大沽河河口底质-水界面耗氧特性

通过对天津大沽河河口底质-水界面细菌降解有机物的耗氧模拟实验,研究河口底质生物耗氧机制。探讨了底质-水界面有机物的耗氧特点及总耗氧中生物耗氧和非生物耗氧所占比例。通过底质-水界面细菌种类、数量、有机物和无机物的含量来判断细菌对含碳源、氮源的有机物的作用。结果表明,20℃,0～14h内,实验期间内平水期河口底质-水界面的生物耗氧和非生物耗氧的比例分别为82 1%和17 9%。异养细菌与有机物及营养盐有较好的正相关关系。河口底质-水界面中异养细菌的组成以革兰氏阳性菌为主,占66%,革兰氏阴性菌只占34%。      

再生水构建水环境中沉水植物附着细菌群落特征

沉水植物茎、叶附着生物膜对再生水构建水环境的水质改善具有重要作用.为探究再生水水质及沉水植物种类对附着生物膜细菌群落特征的影响,选取再生水构建的水环境中不同种类沉水植物为研究对象,采用16S rRNA高通量测序技术对其附着生物膜及周围环境样本的细菌群落结构和功能基因进行分析.结果表明,再生水水体中氮磷营养物质在水环境中得到了20%~35%的吸收利用,下游水体中COD、浊度和色度呈现升高趋势.沉水植物附着生物膜中细菌群落与周围环境（土壤、底泥和水体）及再生水处理厂活性污泥的细菌群落存在差异：在群落多样性上,其丰富度和多样性显著低于土壤和底泥中细菌但高于水中浮游细菌;在群落结构上,其优势菌属及对应相对丰度与其它样本不同,主要优势菌属有鞘氨醇单胞菌属（Sphingomonas）、气单胞菌属（Aeromonas）、假单胞菌属（Pseudomonas）和不动杆菌属（Acinetobacter）,分别占比7%~40%.沉水植物种类和再生水水质（BOD₅、TN、NH₄⁺-N和TP）均会影响植物附着细菌群落,但水质对附着细菌群落的影响大于植物种类,且水质也会影响沉水植物附着细菌群落功能基因相对丰度,氮磷浓度较高区域氮循环与磷循环功能基因相对丰度较高.研究结果为揭示沉水植物附着细菌群落特征和优选沉水植物种植种类提供科学依据.     

反硝化细菌、硝酸钙和锆改性沸石联用对底泥中氮磷迁移转化的影响及硝态氮释放风险评估

本文考察了基于反硝化细菌、硝酸钙和锆改性沸石的组合技术（CN+DB+ZZ）对底泥中氮磷迁移转化的影响,并探讨了该技术的硝态氮释放风险.结果发现,单一的硝酸钙处理（CN）虽然可以有效地抑制底泥中磷的释放,但是会造成上覆水体的氨氮和硝态氮污染.硝酸钙和反硝化细菌联合处理（CN+DB）尽管可以有效地抑制底泥中磷的释放,并降低上覆水体的硝态氮二次污染风险,可是却无法有效地控制底泥中氨氮的释放.硝酸钙和锆改性沸石联合处理（CN+ZZ）虽然可以有效地抑制底泥中磷和氨氮向上覆水体的释放,但却会造成上覆水体硝态氮的二次污染.CN+DB+ZZ组合处理技术不仅可以有效地控制底泥中磷的释放,而且可有效降低底泥中氨氮的释放速率,并且与CN和CN+ZZ技术相比还可降低上覆水的硝态氮二次污染风险.CN+DB+ZZ组合技术对底泥中磷与铁同步释放的抑制、对底泥中氧化还原敏感态磷的削减、以及对底泥吸附磷酸盐和氨氮能力的增强,对于其控制底泥中磷和氨氮的释放是至关重要的.以上结果说明,CN+DB+ZZ组合技术是一种非常有希望的用于控制水体底泥中磷和氨氮释放的方法.     

东海海域表层沉积物中硫酸盐还原菌分布特征研究

利用2011年4、7、8和10月对东海海域4个航次的调查资料,以表层沉积物中硫酸盐还原菌（SRB）为研究对象,针对于SRB所共有的异化型亚硫酸盐还原酶（DSR）中的β亚基基因（dsrB）,通过荧光定量PCR技术对SRB丰度的时空分布特征进行了描述.结果表明,SRB丰度变化范围为1.87×105~4.69×10⁸cells/g,平均值为1.15×10⁸cells/g,且4月SRB丰度最低,7月SRB丰度最高;SRB数量在总细菌中的比例介于0.0039%~1.6176%之间,说明SRB在东海表层沉积物的细菌总量中比例很小;SRB丰度的水平分布特征整体表现为南部海域高于北部海域,长江口及浙闽沿岸泥质区高于非泥质区.此外,SRB丰度与环境因子的相关性分析表明,温度和溶解氧是影响SRB丰度的重要因素.     

生物裂解效应对污泥脱水减量的作用研究

从市政污水处理厂活性污泥中筛选培养出噬菌型细菌,并通过噬菌型细菌的富集与投加,探讨其对市政污泥生物裂解预处理的有效性及其所受环境影响因素与作用规律.研究结果显示,市政污泥中存在具有污泥裂解作用的广谱性噬菌型细菌,宿主菌的投加量可影响噬菌型细菌的富集浓度,但培养基营养物组成浓度对噬菌型细菌的生长增殖无明显影响.当筛选出的噬菌型细菌以10⁶ pfu/mL（污泥）与中性市政污泥在室温下共存培养24h即可有效裂解污泥细胞,污泥比阻与污泥体积较未投加任何菌剂的参照污泥降低了36%和15%.并可导致胞内营养物的释放,污泥液相中总氮与总磷浓度较参照污泥分别上升了245%和242%,溶解性COD与总COD比值（SCOD/TCOD）则提高了195%,但生物裂解作用效果并不与裂解时间始终保持正相关性,最佳污泥生物裂解时间为24h.     

冶炼企业周边农田土壤的多环芳烃污染及其细菌群落效应

多环芳烃是一类持久性有机污染物,进入土壤后可能产生多方面生态效应。为研究多环芳烃对土壤微生物的影响,选取南京某冶炼企业周边农田样品,在分析污染物含量基础上,采用高通量测序、定量PCR等方法综合评价了土壤细菌多样性和组成以及多环芳烃降解细菌丰度等特征。17个土壤样品中,多环芳烃总量为0.25~31.08 mg·kg-1,并具有随污染源距离增加而降低的空间分布特征。与土壤理化性质如p H相比较,多环芳烃污染对土壤细菌的总体多样性和群落组成影响不显著。进一步分析发现多环芳烃与潜在降解微生物的相对丰度和降解功能基因(芳香环羟基化双加氧酶,PAH-RHDα)拷贝数显著正相关。污染较重样品的克隆、测序分析表明,土壤中PAH-RHDα基因主要属于革兰氏阳性细菌nid A3/fad A1类群,且与分支杆菌相关序列较为接近。这些结果综合评价了冶炼企业周边农田土壤多环芳烃污染对微生物群落的影响,提示土壤污染在多环芳烃潜在降解细菌中的富集作用,将为后续污染土壤生物修复提供重要科学依据。     

Effect of salinity on community structure and naphthalene dioxygenase gene diversity of a halophilic bacterial consortium

The aim of this study is to analyze the effect of salinity on polycyclic aromatic hydrocarbons (PAHs) biodegradation, community structure and naphthalene dioxygenase gene (ndo) diversity of a halophilic bacterial consortium with the denaturing gradient gel electrophoresis (DGGE) approach. The consortium was developed from oil-contaminated saline soil after enrichment for six times, using phenanthrene as the substrate. The prominent species in the bacterial consortium at all salinities were identified as halophilic bacteria Halomonas, Alcanivorax, Marinobacter, Idiomarina, Martelella and uncultured bacteria. The predominant microbes gradually changed associating with the saline concentration fluctuations ranging from 0.1% to 25% (w/v). Two ndo alpha subunits were dominant at salinities ranging from 0.1% to 20%, while not been clearly detected at 25% salinity. Consistently, the biodegradation occurred at salinities ranging from 0.1% to 20%, while no at 25% salinity, suggesting the two ndo genes played an important role in the degradation. The phylogenetic analysis revealed that both of the two ndo alpha subunits were related to the classic nah-like gene from Pseudomonas stutzeri AN10 and Pseudomonas aeruginosa PaK1, while one with identity of about 82% and the other one with identity of 90% at amino acid sequence level. We concluded that salinity greatly affected halophilic bacterial community structure and also the functional genes which were more related to biodegradation.

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Microprofiles of activated sludge aggregates using microelectrodes in completely autotrophic nitrogen removal over nitrite (CANON) reactor

Microsensor measurements and fluorescence in situ hybridization (FISH) analysis were combined to investigate the microbial populations and activities in a laboratory-scale sequencing batch reactor (SBR) for completely autotrophic nitrogen removal over nitrite (CANON). Fed with synthetic wastewater rich in ammonia, the SBR removed 82.5±5.4% of influent nitrogen and a maximum nitrogen-removal rate of 0.52 kgN·m⁻³·d⁻¹ was achieved. The FISH analysis revealed that aerobic ammonium-oxidizing bacteria (AerAOB) Nitrosomonas and anaerobic ammonium-oxidizing bacteria (AnAOB) dominated the community. To quantify the microbial activities inside the sludge aggregates, microprofiles were measured using pH, dissolved oxygen (DO), NH4+, NO2− and NO3− microelectrodes. In the outer layer of sludge aggregates (0–700 μm), nitrite-oxidizing bacteria (NOB) showed high activity with 4.1 μmol·cm⁻³·h⁻¹ of maximum nitrate production rate under the condition of DO concentration higher than 3.3 mg·L⁻¹. Maximum AerAOB activity was detected in the middle layer (depths around 1700 μm) where DO concentration was 1.1 mg·L⁻¹. In the inner layer (2200–3500 μm), where DO concentration was below 0.9 mg·L⁻¹, AnAOB activity was detected. We thus showed that information obtained from microscopic views can be helpful in optimizing the SBR performance.