基于PLFA法分析亚硝氮、硝氮和氨氮对厌氧微生物细菌群落的影响

利用磷脂脂肪酸(PLFA)技术研究了亚硝氮、硝氮和氨氮在不同浓度水平下对中温厌氧颗粒污泥中厌氧细菌群落结构的影响.结果表明,高浓度亚硝氮(360 mg·L-1,以N计)、硝氮(300 mg·L-1,以N计)和氨氮(3000 mg·L-1,以N计)使系统COD去除率分别下降至3.49%、10.85%和71.53%,并使污泥表面主要细菌由杆菌变为球状菌;亚硝氮和硝氮引起厌氧细菌和革兰氏阳性菌的含量下降,氨氮引起革兰氏阴性菌的含量下降;PLFA香农-威尔多样性指数随着亚硝氮和硝氮浓度的升高从1.12分别下降至0.96和1.03,氨氮对PLFA香农-威尔多样性指数无明显影响.     

Phosphate removal using aerobic bacterial consortium and pure cultures isolated from activated sludge

In the present study, an attempt is made to evaluate the kinetics of biological phosphate removal using a bacterial consortium of activated sludge, as well as screening for dominant polyphosphates accumulating bacteria. The results showed an efficient phosphate uptake (P < 0.001) of the consortium, with rates related to the initial concentration of both phosphate and carbon sources. Short chain volatile fatty acids presented the suitable substrates for enhanced biological phosphorus removal, of which maximum yield reached 99.23% and 78.51% in basal salt medium supplemented with 0.5% of sodium acetate and lactate respectively. Fifteen phosphate-accumulating bacteria were isolated from the activated sludge and only four isolates were selected and characterized as Pseudomonas aeruginosa AS1, Moraxella lacunata AS2, Acinetobater junii AS3 and Alcaligenes denitrificans AS4. The highest efficiency of phosphate uptake using pure culture was achieved with Ac. junii AS3 (83.36) followed by P. aeruginosa AS1 (81.78%), Al. denitrificans AS4 (76.72%), and M. lacunata AS2 with 50.6%.     

几种可利用多氯联苯和多溴联苯醚植物内生菌的分离及初步鉴定

采用PDA、NA以及高氏一号培养基,分别从毛白杨、绦柳、荻以及二穗短柄草植株内分离、纯化出37株内生菌,其中34株为细菌,3株为放线菌.以分离纯化得到菌株各自对应的营养培养基并添加不同浓度的污染物对获得的37株内生菌进行污染物的耐受筛选,发现24株分别对20 mg·L-1的aroclor1254和BDE209具有一定的耐受性.利用无机盐培养基并添加污染物作为微生物唯一碳源筛选,发现菌株CPY-4和菌株SGL-1可以在只提供4-BDE的无机盐培养基中存活,菌株CPY-4,SGL-1、菌株4、菌株13可以在提供BDE209和aroclor1254的无机盐培养基中存活.形态学观察和16Sr DNA序列同源性分析,初步确定菌株SGL-1为克雷伯氏菌属,菌株CPY-4为地衣芽胞杆菌,菌株4和菌株13为肠杆菌属.     

THE EFFECTS OF SELECTED HERBICIDES ON BACTERIAL POPULATIONS IN AN AQUATIC ENVIRONMENT1

ABSTRACT: Heterotrophic, mesophilic bacteria derived from, and maintained in, a freshwater environment were subjected to three classes of herbicides. Diuron, 2,4,5-Trichlorophenoxyacetic acid, and paraquat, used as water soluble solutions, were added (0.7 and 1.4 mgA) to a laboratory culture of bacteria. Total numbers of the bacteria, representing several genera, were monitored for alteration of numbers and types. Temperature, pH, and dissolved oxygen were monitored and the culture medium was tested for residual herbicide. Alterations observed in populations were considered to result from herbicide additions. Each herbicide tested was found to affect bacterial populations differently. Reasons for observed differences are discussed. In this aquatic environment the herbicides remained qualitatively measurable for periods up to three weeks.     

Quantitative Analyses of Pollution-Indicator and Pathogenic Bacteria in Mumbai Waters from Ballast Water Exchange Perspective

As large numbers of ships either take in ballast from or discharge their ballast into the Mumbai Harbor region, it is pertinent to quantify microorganisms of health concerns and suggest on the suitability of water for ballasting purposes. To meet with this main aim, a selected set of general and pathogenic bacterial groups and their seasonal variations were studied from the Mumbai Harbor area. Sampling was carried out during postmonsoon (November 2001), pre-monsoon (April 2002) and monsoon (October 2002) periods. We quantified total coliforms and eight other groups of known human pathogenic bacteria from water, sediment, marine plant and animal samples collected in and around Mumbai Harbor. When compared with similar studies from other parts/harbors of the world, the Mumbai Harbor area has over 100-times higher levels of coliforms. The serotypes of Escherichia coli O157 and Shigella-Alkaligens Dispar group were abundant throughout the year. Even Vibrio cholerae, V. parahaemolyticus, Salmonella spp., campylobacters and aeromonads were present in large numbers. This comprehensive study, conducted with a view to quantify these bacterial groups and to evaluate the suitability of these waters for ballasting purposes, clearly suggests that both ballasting and deballasting be avoided in this region and alternative procedures should be developed for treating/handling ballast water.     

宏基因组揭示紫色土中邻苯二甲酸酯去除的微生物学机制

邻苯二甲酸酯具有内分泌干扰特性,在环境中持久存在,其去除机制备受关注.为探究邻苯二甲酸酯在紫色农田土壤中的去除机制、关键微生物及功能基因,以邻苯二甲酸二正丁酯（DBP）和邻苯二甲酸二（2-乙基己基）酯（DEHP）为目标污染物,分别设置含量为0、5、10和20 mg·kg^-1的单一污染土壤,避光培养90 d,利用宏基因组技术探究其去除的微生物生态学机制.结果表明,DBP和DEHP在土壤中的降解动态均符合一级动力学降解模型,半衰期介于17.0~38.2 d之间,5 mg·kg^-1的DBP降解速率最快,依次快于10 mg·kg^-1的DEHP、5 mg·kg^-1的DBP、5 mg·kg^-1的DEHP以及20 mg·kg^-1的DBP和DEHP.第7 d和15 d的土壤样品间细菌群落结构存在显著差异,放线菌门相对丰度随时间不断提升,且与DBP或DEHP半衰期呈反比.物种共现网络分析筛选出变形菌门的Pandoraea属为群落中的关键物种,可以用来指示DBP和DEHP污染程度.KEGG功能注释结果表明,Pandoraea在群落中负责酯键水解、苯甲酸降解、群体感应、ABC转运和双组分系统等功能,可以促进邻苯二甲酸酯上下游降解、细胞通讯和其它降解菌的生长繁殖,以此维持群落结构的稳定.由此可见,紫色土中,DBP和DEHP的去除效率取决于初始含量和自身性质,放线菌门的细菌在降解中扮演重要角色,Pandoraea在促进邻苯二甲酸酯降解和调节降解菌群结构和功能的稳定中均发挥关键作用.     

鄱阳湖湿地细菌群落多样性和可培养细菌功能基因丰度

鄱阳湖湿地中蕴含着丰富的微生物资源,参与并维持着湿地生态系统的物质循环和稳定.为探明鄱阳湖湿地土壤细菌分布规律,分析可培养细菌群落结构和功能特征,采用Illumina MiSeq高通量测序技术分析了鄱阳湖湿地土壤全部细菌（AW）和湿地外围周边土壤全部细菌（AS）群落结构多样性特征;又结合可培养方法和qPCR功能基因芯片技术,分析了湿地可培养细菌（CW）和湿地外围周边土壤可培养细菌（CS）功能基因丰度差异.研究发现鄱阳湖湿地土壤细菌多样性低于湿地外围周边土壤细菌,变形菌门和酸杆菌门是两种土壤环境中共有的细菌优势菌门,放线菌门、厚壁菌门、绿弯菌门和热脱硫杆菌门也是存在于两种土壤中的主要菌门,但相对丰度均差异显著;网络分析显示湿地土壤较周边土壤细菌网络结构更简单.采用可培养方法获得的不同采样点土壤细菌群落多样性差异不明显.但湿地土壤可培养细菌功能基因的拷贝数高于周边土壤,其中参与氮循环的UreC基因和参与碳固定的acsE基因在两种土壤可培养细菌中相对丰度较高.研究结果可为深入挖掘调控、维持湿地土壤养分循环和促进鄱阳湖湿地生态系统功能稳定性的潜在有益微生物提供理论依据.     

Pesticides impact on Clavibacter michiganensis ssp. sepedonicus biofilm formation

The effect of various pesticides on the biofilm formation by the phytopathogenic bacterium Clavibacter michiganensis ssp. sepedonicus (Cms), the potato ring rot causative agent, was explored for the first time. Systemic herbicides: 2,4-D, diuron, glyphosate, clopyralid, fluorodifen, as well as the commercial preparations “Lazurite,” “Ridomil Gold,” and the mitochondria inhibiting pesticides analog, sodium monoiodoacetate, were studied. These pesticides' effect on the Cms biofilm formation was shown to be distinct and dependent on the agent under question. Cms biofilm formation was reduced when exposed to sodium monoiodoacetate, as well as “Lazurite” preparation, that could be due to the bactericidal effect of these agents. 2,4-D and “Ridomil Gold” preparation stimulated the biofilm formation. Systemic herbicides diuron, glyphosate, clopyralid, fluorodifen did not exert appreciable influence on the process of bacterial biofilm formation.     

Analysis of bacterial community structures in two sewage treatment plants with different sludge properties and treatment performance by nested PCR-DGGE method

The bacterial community structures in two sewage treatment plants with different processes and performance were investigated by denaturing gradient gel electrophoresis （DGGE） of nested polymerase chain reaction （nested PCR） amplified 16S rRNA gene fragments with group-specific primers. Samples of raw sewage and treated effluents were amplified using the whole-cell PCR method, and the activated sludge samples were amplified using the extracted genomic DNA before the PCR products were loaded on the same DGGE gel for bacterial community analysis. Ammonia-oxidizing bacterial and actinomycetic community analysis were also carried out to investigate the relationship between specific population structures and system or sludge performance. The two plants demonstrated a similarity in bacterial community structures of raw sewage and activated sludge, but they had different effluent populations. Many dominant bacterial populations of raw sewage did not appear in the activated sludge samples, suggesting that the dominant bacterial populations in raw sewage might not play an important role during wastewater treatment. Although the two plants had different sludge properties in terms of settleability and foam forming ability, they demonstrated similar actinomycetic community structures. For activated sludge with bad settling performance, the treated water presented a similar DGGE pattern with that of activated sludge, indicating the nonselective washout of bacteria from the system. The plant with better ammonium removal efficiency showed higher ammonia-oxidizing bacteria species richness. Analysis of sequencing results showed that the major populations in raw sewage were uncultured bacterium, while in activated sludge the predominant populations were beta proteobacteria.     

微生物技术改善河道水质的研究

该课题是探索直接利用人工培养的微生物改善河道水质的研究。在桂林桃花江河道投放反硝化细菌制剂2个月后,水质有了一定程度的改善,COD和TN的浓度降低,但TP和浊度变化幅度不大。受进水水质和低温天气的影响,反硝化细菌制剂的净化作用及反硝化作用不能充分发挥。