Detection, isolation, and identification of cadmium-resistant bacteria based on PCR-DGGE

This study focused on the screening of cadmium-resistant bacterial strains from Pb-Zn tailing. We investigated the diversity of microbial community inhabiting Dong-san-cha Pb-Zn tailing in Beijing, China, by polymerase chain reaction-denaturing gradient gel electrophoresis （DGGE） of 16S rRNA gene of bacterial strain, and found two dominant strains in the DGGE profile. Using special culture media, we isolated two strong cadmium-resistant bacterial strains. On the basis of morphological, physiological, and biochemical characteristics, BIOLOG, and 16S rDNA sequencing, the two strains were identified as Bacillus cereus and Enterobacter cloacae. Minimal inhibitory concentrations （MICs） of heavy metals for the bacteria were determined. E. cloacae showed higher MIC values for heavy metals and a larger range of antibiotic resistance than B. cereus.     

酚降解菌株的分离、鉴定和在含酚废水生物处理中的应用

从石油化工厂的活性污泥和废水混合物中分离到10个降解酚的细菌菌株,经16S rRNA基因序列分析,5个菌株(PD1、PD2、PD6、PD7和PD39)被鉴定为假单胞菌(Pseudomonas sp.),4个菌株(PD4、PD5、PD8和PD9)为不动杆菌(Acinetobacter sp.),1个菌株(PD3)为丛毛单胞菌(Comamonas sp.).对假单胞菌(Pseudomonas sp.)PD39菌株的酚降解特性、最适生长条件、降解底物的范围、儿茶酚1,2-双加氧酶(C12O)和儿茶酚2,3-双加氧酶(C23O)活性、含酚废水的生物处理等,进行了详细研究.结果表明,PD39菌株的最适生长和降解条件是,培养基的起始pH为7.0,培养温度为30℃,酚浓度为800 mg/L,对酚的最大降解浓度为1 200 mg/L.酚浓度为637 mg/L的工业废水经72 h处理酚的去除率达到99.96%.PD39菌株有很好的应用潜力来作含酚废水活性污泥处理系统的强化菌株.     

Bacterial diversity in soils around a lead and zinc mine

Five samples of soil collected from a lead and zinc mine were used to assess the effect of combined contamination of heavy metals on soil bacterial communities using a polyphasic approach including characterization of isolates by culture method, community level catabolic profiling in BIOLOG GN microplates, and genetic community fingerprinting by denaturing gradient gel electrophoresis of 16S rDNA fragments amplified by PCR from community DNA （PCR-DGGE）. The structure of the bacterial community was affected to a certain extent by heavy metals. The PCR-DGGE analysis of 16S rRNA genes showed that there were significant differences in the structure of the microbial community among the soil samples, which were related to the contamination levels. The number of bacteria and the number of denaturing gradient gel electrophoresis （DGGE） bands in the soils increased with increasing distance from the lead and zinc mine tailing, whereas the concentration of lead （Pb） and cadmium （Cd） was decreased. Heavily polluted soils could be characterized by a community that differs from those of lightly polluted soils in richness and structure of dominating bacterial populations. The clustering analysis of the DGGE profiles showed that the bacteria in all the five samples of soil belonged to three clusters. The data from the BIOLOG analysis also showed the same result. This study showed that heavy metal contamination decreased both the biomass and diversity of the bacterial community in soil.     

Phylogenetically clustering of rhizobia by genome structure: application to unclassified Rhizobium

Introduction The rhizobia can change nitrogen in the air into ammonia and provide nitrogenous nutrition to the plant. This will make us independent from the use of nitrogenous fertilizers so as to minimize pollution of water tables, lakes and rivers and r…     

Characterization and phylogenetic analysis of a phenanthrene-degrading strain isolated from oil-contaminated soil

Bacterium strain EVA17 was isolated from an oil-contaminated soil, and identified as Sphingomonas sp.based on analysis of 16S rDNA sequence, cellular fatty acid composition and physiological-chemical tests. The salicylate hydroxylase and catechol 2, 3-dioxygenase (C23O) were detected in cell-free lysates, suggesting a pathway for phenanthrene catabolism via salicylate and catechol. Alignment showed that both of the C23O and GST genes of the strain EVA17 had high similarity with homologues of strains from genus Sphingomonas. The phylogenetic analysis based on 16S rDNA and C23O gene sequence indicated that EVA17 should be classified into genus Sphingomonas, although the two phylogenetic trees were slightly different from each other. The results of co-amplification and sequence determination indicated that GST gene should be located upstream of the C230 gene.     

1株咔唑降解菌的分离、鉴定及其降解特性研究

利用微生物分离技术,从土壤中分离到1株咔唑降解菌KH-6.菌株KH-6能够以咔唑为唯一的碳源和能源良好地生长,同时还可以利用3-甲基咔唑、4-羟基咔唑和2,2′-二羟基联苯生长.经过对其形态特征、生理生化、以及16S rDNA序列分析,初步鉴定为黄杆菌属细菌.该菌株对咔唑的降解性是通过在液体培养基内菌体的增加及底物的减少来证实的,其利用咔唑生长的最适温度和pH分别为30℃和7.5.静止细胞反应试验表明,菌株KH-6除了咔唑以外还可降解其它杂环化合物.咔唑污染土壤中的生物降解实验表明,在灭菌土壤中90%的咔唑被降解,菌株KH-6对土壤中的咔唑有很好的生物降解效果.     

硝酸钙对河流底泥中含硫化合物嗅味原位控制

以硝酸钙作为氧化剂,对深圳河污染底泥典型嗅味物质AVS（酸挥发性硫化物）和GEM（土臭素）及2 MIB（2 甲基异莰醇）进行原位控制. 结果表明,当硝酸钙投加量（以单位质量AVS所需NO3- N质量计）为1.68 g/g,AVS去除率达到92%,且可有效避免硝酸钙向上覆水释放,适合工程应用. 但是,硝酸钙对河流底泥GEM和2 MIB等典型嗅味物质的原位控制效果较差. 硝酸钙能够引起底泥中微生物多样性增加,16S rDNA指纹图谱显示,优势菌株包括去除AVS的脱氮硫杆菌(Thiobacillus thioparus),反硝化细菌(Simplicispira sp.和Rhodanobacter sp).以及降解有机物的菌株(Simplicispira sp.、Lysobacter sp.、Pseudoxanthomonas sp.)等.      

新乡市镉污染土壤细菌群落组成及其对镉固定效果

重金属污染土壤中具有丰富的微生物群落组成,为生物修复提供了微生物资源.本研究以新乡市某电池厂周边Cd污染土壤为研究对象,采用高通量测序和平板分离方法分析其细菌群落组成.传统培养方法表明新乡市重金属污染土壤细菌由厚壁菌门、放线菌门、变形菌门和拟杆菌门这4个门,芽孢杆菌属、节杆菌属和根瘤菌属等30个属组成;高通量测序表明,其由变形菌门、放线菌门和酸杆菌门等25门及400属组成.相较于培养方法,高通量测序群落组成更为丰富.基于高通量测序的分子生态网路分析表明其关键细菌分别为Arthrobacter、Marmoricola、Nocardioides、Ferruginibacter、Flavitalea、Nitrospira和Lysobacter等组成.对分离的159株可培养菌株进行Cd摇瓶吸附实验,结果表明Aneurinibacillus、Arthrobacter和Bacillus等11个属的30株菌具有较好地固定效果.效果验证实验表明,Ochrobactrum sp. 1-6、Bacillus sp.2-11和Pseudomonas sp.1-9等6株高效菌株能提高青菜(鸡毛菜)生物量、降低...     

1株高效产絮凝剂菌株的分离及其在冷轧含油废水处理中的应用

从土壤中分离出1株高效产絮凝剂菌株T-3,经形态学特征分析、生理生化实验及16S rDNA序列分析,鉴定为克雷伯氏菌（Klebsiella sp.）.研究了该菌株的培养条件,如pH值、温度、碳源、氮源对产絮凝剂性能的影响.结果表明,该菌株对碳源、氮源有较普遍的适应性;初始pH值为9,培养温度为25℃时,产絮凝剂性能较好...     

菌株EA41对醋酸酯类化合物的生物降解

利用醋酸乙酯为能源和碳源从土壤中分离到一株菌。在生理生化试验的基础上,利用16S rDNA的方法鉴定这株菌为假单胞菌属细菌。底物特异性试验表明,假单胞菌EA41菌株可代谢多种醋酸酯类化合物。休眠细胞反应试验表明,这株菌的水解酶能水解口香糖的主要成分聚醋酸乙烯酯产生醋酸。     

苯并[a]芘降解菌SL-1的筛选、鉴定及其降解特性

从土壤中筛选出一株以苯并[a]芘为唯一碳源的高效降解菌SL-1,经形态观察、生理生化分析以及16SrDNA基因序列分析,该菌株鉴定为假单胞菌（Pseudomonassp．）。28℃震荡培养10d后,菌株SL-1对5mg·L^-1苯并[a]芘的降解率为35．7％;同时测定该降解菌的生长曲线、降解率变化曲线、最佳培养条件;土壤修复试验表明,42d时,降解菌SL-1对苯并芘的降解率达到76．3％。     

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.     

解淀粉乳酸细菌在厨余垃圾乳酸发酵中的应用

研究了水解淀粉乳酸细菌对厨余垃圾发酵生产乳酸的强化作用.从厌氧发酵的厨余垃圾中分离到6株水解淀粉的乳酸细菌,其中菌株FH164具有最高的淀粉降解率和乳酸产量.在pH5.5～6.0条件下,经48h发酵,菌株FH164从40.50g·L^-1的可溶性淀粉产生32.67g·L^-1的乳酸.根据形态、生理生化特征和16SrDNA序列分析结果,可将菌株FH164鉴定为链球菌属(Streptococcus sp.)细菌.接种菌株FH164可强化厨余垃圾的乳酸发酵,采用垃圾不灭菌的开放式发酵,菌株FH164可得到28.23g·L^-1的乳酸,比自然发酵(不接种的对照)的乳酸浓度高19.2%.     

嗜热溶胞菌SY-14的基本特性研究

对从污泥堆肥中分离得到的具有溶胞能力的嗜热产芽孢杆菌SY-14进行了初步鉴定,并对其生长、产酶及溶胞特性进行了研究.根据菌株形态及16S rDNA测序结果,SY-14为土芽胞杆菌属(Geobacillus sp.).SY-14最适生长温度60℃左右,最适生长pH为6.0～7.0.SY-14培养上清液对完整和加热灭活后的微生物细胞都具有溶胞能力,6 h的细胞溶解率分别可达到70%和85%,上清液经过热处理后溶胞能力明显下降,表明溶胞能力主要来自于酶的作用.间歇培养过程中,SY-14培养上清液溶胞活性在对数生长期快速升高,进入稳定期前后达到最大,之后快速下降.SY-14培养上清液对受试的5株革兰氏阴性菌及部分革兰氏阳性菌都具有溶胞能力.     

4株邻苯二甲酸二丁酯降解菌的分离鉴定及其相关降解基因的克隆

从土壤中分离纯化出4株能降解邻苯二甲酸二丁酯(DBP)的菌株,分别命名为JDC-1、JDC-8、JDC-9、JDC-12,并对其进行了形态学、生理生化及分子生物学鉴定.菌株革兰氏染色阳性,16S rDNA序列分析显示4株菌均与节杆菌属(Arthrobacter sp.)有99%以上的序列相似性,初步判断这4株菌为Arthrobacter sp..通过PCR扩增及克隆,均获得了1个约900 bp的DNA片段,测序结果显示该片段与Arthrobacter keyseri的邻苯二甲酸3,4-双加氧酶基因的核苷酸序列相似性为96%以上.对4株菌的最适生长条件及对DBP的降解能力进行了分析,结果显示,4株菌的最适生长条件为pH 7.0~8.5,温度30~35℃.以DBP作为目标测试物,在适宜条件下测试了4株菌的降解能力,显示这4株菌均为高效降解菌,效率最高的JDC-1能在28 h内将500 mg/L的DBP降解完全,最慢的JDC-8经40 h能将500 mg/L的DBP降解完全,本研究对于DBP降解机制的研究及微生物资源的开发都具有重要意义.     

降解芘的分枝杆菌M11的分离鉴定和降解特性

从多环芳烃污染的土壤中分离到1株能高效降解四环芳烃芘的放线菌M11,经形态观察、生理生化和16S rDNA鉴定,属于分枝杆菌属(Mycobacterium sp.).菌株M11能以菲、蒽、荧蒽和芘为唯一碳源生长,在含芘50、100和200 mg/L的无机盐液体培养基中培养16 d降解率分别达到76.9%、91.8%和79.23%.菌株M11对芘的降解具有较广泛的pH范围,在芘浓度100 mg/L,pH为5～9的液体条件下,均可生长.根据已报道的芘降解菌的双加氧酶同源序列设计引物,PCR扩增出编码双加氧酶大亚基和小亚基的基因片段,序列分析表明与已知降解芘的分枝杆菌的双加氧酶基因具有高度同源性.     

三株菲降解细菌的分离、鉴定及降解特性的研究

从石油污染土壤中分离纯化到 3株能以菲为唯一碳源和能源生长的革兰氏阴性杆菌、好氧、不形成芽孢、端生或侧生单根鞭毛 ,分别命名为菌株ZX4、ZX6和EVA17.据部分片段长度的 16SrDNA序列比对分析和生理生化试验结果 ,确认此 3株菌均为鞘氨醇单胞菌属 (Sphingomonas)细菌 ,菌株ZX4为少动鞘氨醇单胞菌 (S .paucimobilis) ,菌株ZX6为溶芳烃鞘氨醇单胞菌 (S .aromaticivorans) ,菌株EVA17尚不能确定 .分析了各菌株在系统发育中的分类地位 .菌株ZX4在 14d内对含量为 10 0 0mg·L-1的菲降解率可达 98 74%.不同底物实验结果表明 ,菌株EVA17可能同时拥有两条菲降解途径 .     

厌氧/好氧交替快速筛选聚磷菌及其生理特性的研究

通过厌氧/好氧交替的平板筛选方法,快速的筛选出除磷率高于60%的高效聚磷菌15株,并对其进行16S rDNA和生理生化测定,除2株为芽孢杆菌外,其余均为γ变形菌纲,主要以Klebsiella sp.和Pseudomonas sp.为主.菌种除磷实验表明,聚磷菌除磷能力越高,发酵终点pH值越高.聚磷菌反投加实验表明,与活性污泥52%的除磷率相比,投加后除磷率可达到73.3%,可以有效提高活性污泥的除磷能力.     

1,2,4-三氯苯矿化菌的鉴定与功能分析

采用同位素示踪法从氯苯污染土壤中筛选到能高效降解1,2,4-三氯苯(1,2,4-TCB)为CO₂的细菌E3和F2,根据其表型特征、16S rDNA序列和荧光原位杂交(FISH)分析,鉴定E3和F2为博德特氏菌（Bordetella sp.）.E3和F2的16S rDNA序列相似性为100%;它们与菌株Bordetella sp. QJ2-5同源性最高,与Bordetella petrii(GDH030510)最接近,同源性分别为100%和99.4%.E3和F2能在含1,2,4-TCB的无机盐培养基中彻底降解1,2,4-TCB生成CO₂,30 d降解率达到90%以上,其中58%和46%矿化为CO₂,同时形成一定量的微生物生物量.     

青藏铁路沿线土壤可培养微生物种群多样性分析

选择青藏铁路沿线不同海拔高度的10个地点采集土壤样品,采用3种不同的培养基分离培养其中的微生物;提取各土壤样品可培养微生物菌体基因组DNA,进行16S rDNA的PCR扩增,对可培养微生物种群多样性进行分析.结果表明,该地区可培养微生物的数量为1×106～8×106 CFU/g.16S rDNA序列分析发现,分离培养的微生物主要包括厚壁菌门(Firmicutes)、放线菌门(Actinobacteria)和变形菌门γ亚群(Gammaproteobacteria) 3个类群,分别占菌株总数的53.1%,29.6%和16.0%.其中,Bacillus sp.为优势菌属,占菌株总数的29.6%;Bacillus simplex为优势种,占菌株总数的12.3%.