一株DEHP降解菌的筛选和分子鉴定

采用富集培养法从污水厂活性污泥中分离得到一株能以邻苯二甲酸二(2-乙基己基)酯(DEHP)为唯一碳源的细菌。该菌在48 h内可将DEHP浓度从20 mg/L降解到2 mg/L。对其进行了生理生化试验和分子生物学鉴定。结果表明,该菌株呈丝状,革兰氏阳性,好氧,能在45℃下生长。该菌株与长孢糖丝菌模式菌Saccharothrix longispora DSM 43749在同一分枝上,同源性为98.9%;与糖丝菌属(Saccharothrixsp)内其他种同源性均在97.0%以上。     

一株产黑色素海洋细菌的筛选、鉴定及发酵条件优化

从山东近岸海域海水中分离得到一株产黑色素细菌Sl,通过对S1菌株的菌落形态观察、生理生化性质检验及16SrDNA序列测定,初步鉴定为明亮发光杆菌(Photobacterium phosphoreum).为了提高S1菌株黑色素产量,采用单因素实验设计对S1菌株进行发酵条件优化,结果表明当发酵条件为L-酪氨酸0.75 g/...     

一株产黑褐色色素海洋放线菌的研究

从舟山群岛潮间带海域中分离到一批产色素的海洋放线菌,用滤纸片法进行抑菌试验,并对目标菌株进行鉴定,然后对其所产色素进行粗提,测色素的最大吸收波长,研究理化因子对其影响.结果表明筛选到产黑褐色色素的菌株B2,其对Vibrio parahaemolyticus和Escherichia coli具有较好的抑制作用,色素初步研...     

Taxonomy of oxalotrophic <Emphasis Type="Italic">Methylobacterium</Emphasis> strains

Most of the oxalotrophic bacteria are facultative methylotrophs and play important ecological roles in soil fertility and cycling of elements. This study gives a detailed picture of the taxonomy and diversity of these bacteria and provides new information about the taxonomical variability within the genus Methylobacterium. Twelve mesophilic, pink-pigmented, and facultatively methylotrophic oxalate-oxidizing strains were included in this work that had been previously isolated from the soil and some plant tissues by the potassium oxalate enrichment method. The isolates were characterized using biochemical tests, cellular lipid profiles, spectral characteristics of carotenoid pigments, G+C content of the DNA, and 16S rDNA sequencing. The taxonomic similarities among the strains were analyzed using the simple matching (S _SM) and Jaccard (S _J) coefficients, and the UPGMA clustering algorithm. The phylogenetic position of the strains was inferred by the neighbor-joining method on the basis of the 16S rDNA sequences. All isolates were Gram-negative, facultatively methylotrophic, oxidase and catalase positive, and required no growth factors. Based on the results of numerical taxonomy, the strains formed four closely related clusters sharing ≥85% similarity. Analysis of the 16S rDNA sequences demonstrated that oxalotrophic, pink-pigmented, and facultatively methylotrophic strains could be identified as members of the genus Methylobacterium. Except for M. variabile and M. aquaticum, all of the Methylobacterium type strains tested had the ability of oxalate utilization. Our results indicate that the capability of oxalate utilization seems to be an uncommon trait and could be used as a valuable taxonomic criterion for differentiation of Methylobacterium species.     

Purification of total DNA extracted from activated sludge

Purification of the total DNA extracted from activated sludge samples was studied. The effects of extraction buffers and lysis treatments (lysozyme, sodium dodecyl sulfate (SDS), sonication, mechanical mill and thermal shock) on yield and purity of the total DNA extracted from activated sludge were investigated. It was found that SDS and mechanical mill were the most effective ways for cell lysis, and both gave the highest DNA yields, while by SDS and thermal shock, the purest DNA extract could be obtained. The combination of SDS with other lysis treatment, such as sonication and thermal shock, could apparently increase the DNA yields but also result in severe shearing. For the purification of the crude DNA extract, polyvinyl polypyrrolidone was used for the removal of humic contaminants. Cetyltrimethyl ammonium bromide, potassium acetate and phenol/chloroform were used to remove proteins and polysaccharides from crude DNA. Crude DNA was further purified by isopropanol precipitation. Thus, a suitable protocol was proposed for DNA extraction, yielding about 49.9 mg (total DNA)/g volatile suspended solids, and the DNA extracts were successfully used in PCR amplifications for 16S rDNA and 16S rDNA V3 region. The PCR products of 16S rDNA V3 region allowed the DGGE analysis (denatured gradient gel electrophoresis) to be possible.     

Eubacteria and Archaea community of simultaneous methanogenesis and denitrification granular sludge

Based on the successful performance of a lab-scale upflow anaerobic sludge blanket (UASB) reactor with the capacity of simultaneous methanogenesis and denitrification (SMD), the specific phylogenetic groups and community structure of microbes in the SMD granule in the UASB reactor were investigated by the construction of the Eubacteria and Archaea 16S rDNA clone libraries, fragment length polymorphism, and sequence blast. Real time quantitative-polymerase chain reaction (RTQ-PCR) technique was used to quantify the contents of Eubacteria and Archaea in the SMD granule. The contents of some special predominant methanogens were also investigated. The results indicated that the Methanosaeta and Methanobacteria were the predominant methanogens in all Archaea in the SMD granule, with contents of 71. 59% and 22. 73% in all 88 random Archaea clones, respectively. The diversity of Eubacteria was much more complex than that of Archaea. The low GC positive gram bacteria and Б-Protebacteria were the main predominant Eubacteria species in SMD granule, their contents were 49. 62% and 12. 03% in all 133 random Eubacteria clones respectively. The results of RTQ-PCR indicated that the content of Archaea was less than Eubacteria, the Archaea content in total microorganisms in SMD granule was about 27. 6%.     

啤酒废水处理系统高效菌株的分离筛选与分析

从啤酒废水处理系统SBR池中取得的水样,分离纯化得到菌株35株,分别以进站原水和水解酸化池出水作为污水培养基,进行COD降解试验,复筛后分别得到3株COD降解率高且降解效果稳定的细菌12#,15#,23#与15#,25#,31#。将高效菌株与SBR池的活性污泥混合,考察其对啤酒废水的处理效果,结果表明,23#与活性污泥交互作用时在原水污水培养基中摇床培养16h COD降解率达到75.13%;25#与活性污泥交互作用处理水解酸化池出水120h时降解率达到82.40%。对4株高效菌株15#,23#,25#,31#进行16S rDNA序列测定,将所得序列与GenBank中已登录的相近细菌种群的DNA序列对比,得到高度同源性的细菌菌种。     

不同空间位点沉积物理化性质与微生物多样性垂向分布规律

运用化学分析方法和PCR-DGGE技术从沉积物化学及基因角度对太湖不同空间位点沉积柱理化性质(pH、Eh)、营养盐(TN、TP、OM)及微生物多样性的垂向分布及相关性进行研究.结果表明,沉积物Eh在水-土界面处于轻度还原状态,表层下,随沉积深度的增加迅速下降,还原性逐渐增强.沉积物pH在整个剖面上变化幅度不大,在7.2～7.8变动.沉积物中含有丰富的营养盐,总氮和总磷最高含量分别为 2.436 mg/g和 0.731 mg/g,其剖面分布特征表明,沉积物表层总氮和总磷含量远高于底层,其含量随深度增加而降低.沉积物有机质含量在15 cm以上变化幅度较大,随着沉积深度的增加,有机质含量明显减少.沉积物中微生物群落结构显示了明显的空间分布多样性差异.三位点TN与OM之间均存在显著的相关性,但理化指标与营养盐以及微生物群落多样性指标之间不存在显著的相关性.     

一株嗜碱产甲烷八叠球菌的分离鉴定与系统发育分析

在15℃条件下用产甲烷菌培养基对采自四川省红原县的牦牛粪进行富集培养,采用Hungate厌氧操作技术从富集培养物中分离得到一株在8~45℃范围生长、最适生长pH为8.5的嗜碱产甲烷菌T13.该菌株革兰氏染色阳性,细胞聚集体,在液体培养基中为肉眼可见的颗粒状物,在固体培养基上菌落为淡黄色桑葚状;可利用甲醇、乙酸盐和甲胺作为唯一碳源生长;对氯霉素和庆大霉素敏感;生长pH范围为6.5~9.5;最适NaCl浓度为0~0.15 mol L-1;最适生长温度为30℃.形态和生理生化特征以及16S rDNA序列分析表明菌株T13为梅氏产甲烷八叠球菌(Methanosarcina mazei).由于该菌最适生长pH为8.5,所以初步认为菌株T13是一株梅氏产甲烷八叠球菌的新菌株.     

抚顺西露天油页岩可培养微生物的分离与鉴定

为了解油页岩中可培养微生物的多样性,利用稀释平板培养法对我国抚顺盆地西露天组油页岩中可培养微生物进行分离纯化,并对所获菌株进行16S rDNA(细菌)和rDNA ITS(真菌)序列测定和系统发育树分析.以营养琼脂培养基分离到8株细菌,以马铃薯葡萄糖琼脂培养基分离到4株真菌.16S rDNA序列测定结果表明细菌菌株属于芽孢杆菌属(Bacillus),赖氨酸芽孢杆菌属(Lysinibacillus),短杆菌属(Brevibacillus),类芽胞杆菌属(Paenibacillus)和红球菌属(Rhodococcus); rDNA ITS序列测定结果显示真菌菌株分别属于芽枝霉属(Cladosporium),曲霉属(Aspergillus),犁头霉属(Absidia)和正青霉属(Eupenicillium). 2株细菌(ZK4和ZK5)能够在以油页岩为唯一碳源和氮源的固体培养基上连续生长5代以上,且长势良好, 可作为油页岩生物转化的潜力菌株进行深入研究.