氧化硫硫杆菌质粒的分离

用碱法对分离到的7株氧化硫硫杆菌进行质粒检测,在6株中分离到了18个质粒,质粒DNA的分子量为1.3×10~4-98×10~6。     

猪胸膜肺炎放线杆菌ApxIIA基因的序列分析及其B细胞表位预测

为了进一步研究猪胸膜肺炎放线杆菌(Actionobacillus pleuropneumoniae,App)ApxIIA基因的结构和功能,选取GenBank中6株不同App分离株,采用生物信息学方法对其ApxIIA基因及其编码氨基酸序列进行同源性比对,并选择其中AY232288.1菌株(湖北分离株)ApxIIA基因为研究对象,分析该基因所编码蛋白质的理化性质,并对其可能形成的二级结构及B细胞表位进行预测和分析.结果表明,6株不同App分离株ApxIIA基因核苷酸及氨基酸序列同源性分别为65.56﹪和88.42﹪,在AY232288.1菌株ApxIIA蛋白的肽链中,745～751和801～807区段可能是其B细胞表位优势区.本研究首次利用生物信息学方法对ApxIIA基因及其蛋白质结构进行了分析和预测,为ApxIIA基因功能的深入研究及APP多表位疫苗的设计奠定了基础.     

罗伊氏乳杆菌肠道分离株的遗传多样性研究

动物体内定殖着丰富且多样的细菌,其对宿主的健康发挥着举足轻重的作用。罗伊氏乳杆菌(Lactobacillus reuteri)是存在于动物肠道中的益生菌,是研究肠道菌群与宿主进化关系的模式菌种。【目的】以下载自NCBI的分离自猪、家禽类、人类、啮齿类动物的116株L.reuteri和分离自内蒙古锡林郭勒牛、羊和马肠道中的16株L. reuteri为研究对象,解析L. reuteri不同分离株的遗传多样性和宿主特异性,为L. reuteri的开发利用提供理论依据。【方法】利用MLST技术,以ddl、pkt、leu S、gyr B、dlt A、rpo A、rec A共7个看家基因为研究靶点,对L.reuteri分离株遗传多样性进行研究,推演分离株与宿主生境的进化关系。【结果】132株L. reuteri共划分为63个序列型,6个克隆复合体。等位基因序列重组分析发现,在L.reuteri的进化中发生了个别的重组事件,eBURST、MSTree分析表明不同分离源的L. reuteri分离株经历了不同的进化过程,系统发育分析表明132株L. reuteri来自5个Clusters且与分离源表现出较强的相关性。【结论】本研究利用MLST技术完成了132株L. reuteri肠道分离株的遗传多样性分析,利用MSTree、系统发育等群体结构分析,发现不同分离源菌株有着高度的宿主特异性,表明L. reuteri为适应不同生存环境经历了不同的进化过程。     

不同分离源植物乳杆菌的群体基因组分析

【背景】植物乳杆菌(Lactobacillus plantarum)广泛存在于植物、乳制品、肉制品、哺乳动物和昆虫的肠道等多种生态环境中。【目的】探究不同分离源L. plantarum基因组与其所在环境是否存在潜在的联系。【方法】利用比较基因组学对126株分离自植物、乳制品、肉制品、果蝇及哺乳动物肠道和口腔等部位的L. plantarum菌株基因组进行系统发育分析和功能基因组分析,解析不同分离源菌株间的亲缘关系和进化历程。【结果】果蝇分离株的基因组大小显著高于植物、哺乳动物肠道、肉制品和乳制品分离株(P0.05),植物和哺乳动物肠道、口腔等部位与肉制品分离株的基因组大小和编码基因数量无显著差异(P0.05)。基于单拷贝基因串联和核心基因系统发育树分析均发现,果蝇分离株和乳制品分离株分别集中聚集分布在某一分支中,其余分离源均匀分布在各个分支中。附属基因分析结果与系统发育树分析结果一致。功能基因注释结果发现,果蝇分离株的环境特异性基因参与低聚果糖和几丁质代谢,乳制品分离株的环境特异性基因参与mazEF毒素-抗毒素系统和CRISPR系统。【结论】植物乳杆菌分离株为适应较为独特的果蝇和乳制品生境而发生了适应性进化。本研究为植物乳杆菌适应性进化提供了新见解,同时为解析菌株的进化历程提供了理论基础。     

不同风化程度钾长石表面矿物分解细菌的筛选及遗传多样性

【目的】不同风化程度钾长石表面矿物分解细菌生物多样性研究将有助于了解矿物生物风化、生物成矿和土壤形成的演化规律和机理。【方法】采用纯培养法自南平钾矿区高、中、低风化度钾长石以及矿区土壤样品中分离矿物分解细菌,通过摇瓶释硅实验比较不同菌株分解矿物能力,采用16SrDNA限制性酶切多态性分析(Amplified rDNA Restriction Analysis,ARDRA)研究了供试菌株的遗传多样性。【结果】分离筛选到35株生长良好的矿物分解细菌,与对照相比,接菌处理发酵液中有效硅增加了101～206%;所有供试菌株可分为11个OTU,分别属于5个门,6个科,7个属。多数菌株(74%)属于γ-变形杆菌纲(γ-Proteobacteria)。泛菌属(Pantoea),沙雷氏菌属(Serratia),假单胞菌属(Pseudomonas)为优势种群。【结论】南平钾矿区矿物分解细菌具有丰富的微生物种群多样性,且γ-变形杆菌纲(γ-Proteobacteria)细菌在钾长石风化过程中可能起了重要的作用。     

安徽PRSS发病猪副猪嗜血杆菌的分离鉴定及其耐药性分析

目的了解猪繁殖与呼吸综合征（PRRS）发病猪继发感染副猪嗜血杆菌（HPS）的情况及分离菌株的药物感受性。方法应用细菌分离培养、形态学检查、生化试验和PCR技术,对2007年1月至2008年12月从安徽不同地区采集的146头份PRRS发病猪的病料进行HPS检测,并采用标准K-B纸片法对分离菌株进行14种抗菌药物敏感试验。结果分离鉴定出12株HPS,检出率为8.22%（12/146）;12株HPS对氯霉素100%敏感,环丙沙星为91.7%,阿莫西林和新霉素为83.3%,对罗红霉素100%耐药,阿米卡星为83.3%。结论安徽省不同地区PRRS感染猪群中均存在程度不同的HPS感染,各地区HPS分离株表现出形态上的变化特征和一致的生化特性,且有对临床常用抗菌药物耐药性增强的趋势。     

Viili乳制品中干酪乳杆菌的分离鉴定

从引进Viili乳制品中筛选、鉴定出3种优良乳酸菌。采用平板分离法从Viili乳制品中分离3株乳酸菌,通过表型1、6S rRNA的PCR扩增、克隆、测序鉴定。3株乳酸菌的表型鉴定结果符合伯杰细菌鉴定手册中乳杆菌属的干酪乳杆菌,16S rRNA序列同源性分析结果表明3株分离菌与干酪乳杆菌的同源性分别为99.93%、100.00%9、9.78%,从Viili乳制品分离到3株干酪乳杆菌。     

直链烷基苯磺酸钠(LAS)降解菌的筛选及其降解特性的初步研究

从洗衣粉生产废水中分离到一株高效降解LAS(Linear Sodium A]kylbenzenesulfonate)菌GZ6,经初步鉴定其为杰氏棒杆菌(Corynebactenunjeikeiun GZ6).该菌株最高可以降解700mg/L左右的LAS,降解LAS的最适pH值和温度分别为7.0和30℃,最适LAS浓度为400 mg/L,所需时间为24h,降解率可达98.7%.实验还表明一些重金属离子如-Hg2+、Co2+、Cd2+等对该菌株降解LAS有不同程度的抑制作用.     

多耐药鲍曼不动杆菌噬茵体IME—AB6的分离、生物特性及保存方法研究

目的：分离并鉴定一株多耐药鲍曼不动杆菌噬菌体,对其生物特性进行研究,为治疗多耐药鲍曼不动杆菌感染提供新的方法和实验依据。方法：以一株多耐药不动杆菌AB6为宿主菌,从医院废水中分离出噬菌体;用聚乙二醇6000对噬菌体进行浓缩和纯化,并就噬菌体的形态、一步生长曲线、裂解特性和不同保存条件对噬菌体活性的影响进行初步研究。结果：分离出一株噬菌体IME-AB6,电镜显示为肌尾噬菌体,其潜伏期为10 min,爆发期为40 min,爆发量160 pfu/cell;IME-AB6能迅速使菌液变清晰,温度灵敏性强,并且发现在4℃保存是一种方便高效的方法。结论：噬菌体IME-AB6是一株新的有独特特点的噬菌体,用聚乙二醇能很好地提高滴度,其潜伏期和爆发期短且杀菌效能强,性能稳定易保存,这对于噬菌体制剂的开发和耐药性鲍曼不动杆菌的防治具有潜在应用价值。     

安徽部分地区猪丹毒杆菌的分离鉴定及生物学特性研究

【目的】对安徽省8个不同地区猪场临床疑似猪丹毒病/死猪进行细菌分离鉴定,并研究其生物学特性。【方法】通过形态学及培养特性观察、生化试验、PCR方法对菌株进行鉴定,并进行药物感受性实验及免疫保护实验。【结果】共分离到29株猪丹毒杆菌,源自8个地区的猪丹毒杆菌分离菌具有较一致的形态特征和相似的生化特性。对29株猪丹毒杆菌进行18种常用抗菌药物的药敏试验,结果显示分离菌对氨苄西林、头孢曲松敏感率均达100%,其次是青霉素93%、红霉素89.7%和头孢噻肟75.9%,对其他13种药物则表现不同程度的耐药性。8株不同地区猪丹毒杆菌分离菌的LD50在(14.30?2.36)×102 CFU/mL之间,显示分离菌对小鼠均具有较强的致病力。商品化猪丹毒G4T10株弱毒疫苗2次颈部皮下免疫小鼠后,分别用剂量为100 LD50的8株猪丹毒杆菌分离菌腹腔攻毒小鼠,免疫保护率为100%。【结论】安徽地区猪丹毒发生有上升趋势,不同地区的猪丹毒杆菌分离菌具有较为一致的生物学特性,青霉素类和头孢类抗菌药物有显著疗效,使用猪丹毒G4T10株弱毒疫苗可产生有效的免疫保护力。     

Bacterial Diversity in Linglong Gold Mine,China

Bacteria have been actively regulating cycles of various elements in the environment. To explore the potential bacterial role in gold biogeochemical cycling, this study analyzed the bacterial diversity of mine rock (MR) and surface soil (SS) samples from Linglong gold mine using 16S rRNA gene clone library analysis and cultivation method. From MR, 24 operational taxonomic units (OTUs) were identified from MR, covering 3 phyla and 18 genera. Meanwhile, 24 OTUs were identified from SS, including 4 phyla and 18 genera. Compared with 16S rRNA gene clone library analysis, 28 aerobic and 34 anaerobic isolates were obtained, whereas 26 aerobic and 71 anaerobic strains were isolated from SS. The cultivable bacteria were affiliated with Firmicutes, Proteobacteria and Actinobacteria phyla, and dominated by Firmicutes. These results underscore the high level of bacterial diversity in the gold mine. Our study provides information on the microbial diversity in Linglong gold mine and sheds light on the existence and potential function of bacteria in the gold biogeochemical cycling.     

Molecular characterization of Acidithiobacillus ferrooxidans and A. thiooxidans strains isolated from mine wastes in Brazil

Nineteen strains of Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans, including 12 strains isolated from coal, copper, gold and uranium mines in Brazil, strains isolated from similar sources in other countries and the type strains of the two species were characterized together with the type strain of A. caldus by using a combination of molecular systematic methods, namely ribotyping, BOX- and ERIC-PCR and DNA-DNA hybridization assays. Data derived from the molecular fingerprinting analyses showed that the tested strains encompassed a high degree of genetic variability. Two of the Brazilian A. ferrooxidans organisms (strains SSP and PCE) isolated from acid coal mine waste and uranium mine effluent, respectively, and A. thiooxidans strain DAMS, isolated from uranium mine effluent, were the most genetically divergent organisms. The DNA-DNA hybridization data did not support the allocation of Acidithiobacillus strain SSP to the A. ferrooxidans genomic species, as it shared only just over 40% DNA relatedness with the type strain of the species. Acidithiobacillus strain SSP was not clearly related to A. ferrooxidans in the 16S rDNA tree.     

Archaeal diversity in waters from deep South African gold mines.

A culture-independent molecular analysis of archaeal communities in waters collected from deep South African gold mines was performed by performing a PCR-mediated terminal restriction fragment length polymorphism (T-RFLP) analysis of rRNA genes (rDNA) in conjunction with a sequencing analysis of archaeal rDNA clone libraries. The water samples used represented various environments, including deep fissure water, mine service water, and water from an overlying dolomite aquifer. T-RFLP analysis revealed that the ribotype distribution of archaea varied with the source of water. The archaeal communities in the deep gold mine environments exhibited great phylogenetic diversity; the majority of the members were most closely related to uncultivated species. Some archaeal rDNA clones obtained from mine service water and dolomite aquifer water samples were most closely related to environmental rDNA clones from surface soil (soil clones) and marine environments (marine group I [MGI]). Other clones exhibited intermediate phylogenetic affiliation between soil clones and MGI in the Crenarchaeota. Fissure water samples, derived from active or dormant geothermal environments, yielded archaeal sequences that exhibited novel phylogeny, including a novel lineage of Euryarchaeota. These results suggest that deep South African gold mines harbor novel archaeal communities distinct from those observed in other environments. Based on the phylogenetic analysis of archaeal strains and rDNA clones, including the newly discovered archaeal rDNA clones, the evolutionary relationship and the phylogenetic organization of the domain Archaea are reevaluated.     

Isolation of Arsenic Resistant and Arsenopyrite Oxidizing Acidithiobacillus Species from pH Neutral Colombian Mine Effluents

Abstract

Inactive mines provide a great source of bacterial diversity for studying acidophilic communities and their biotechnological applications, but prospecting of these anthropogenic environments in Colombia has been limited. Conventional microbiological methods were used to isolate acidophilic bacterial strains from effluents emanating from the Colombian gold mine ‘El Zancudo’ (Titiribí, Antioquia). Despite the drainage waters having circumneutral pH, all of the isolated strains were phylogenetically related to the extreme acidophile Acidithiobacillus genus. However, based upon 16S rRNA gene sequences the mesophilic sulfur-oxidizing indigenous strains could not be assigned to a species. Pure cultures were selected by screening in medium with soluble inorganic arsenic (III) and their mineral-oxidative activity was evaluated at 30?°C in Erlenmeyer flasks with arsenopyrite ore under rotary shaking conditions. The indigenous strains were able to catalyze arsenopyrite oxidation in a mixed culture with a pulp density of 10%, maintaining their growth in the presence of >80?mM leached arsenic. This research provides information regarding the isolation of arsenic resistant bacterial communities from neutral effluents from El Zancudo mine and the possibility of the isolated strains to be useful in the biooxidation pretreatment of refractory gold-bearing arsenopyrite ores and concentrates.     

Archaeal Diversity in Waters from Deep South African Gold Mines

A culture-independent molecular analysis of archaeal communities in waters collected from deep South African gold mines was performed by performing a PCR-mediated terminal restriction fragment length polymorphism (T-RFLP) analysis of rRNA genes (rDNA) in conjunction with a sequencing analysis of archaeal rDNA clone libraries. The water samples used represented various environments, including deep fissure water, mine service water, and water from an overlying dolomite aquifer. T-RFLP analysis revealed that the ribotype distribution of archaea varied with the source of water. The archaeal communities in the deep gold mine environments exhibited great phylogenetic diversity; the majority of the members were most closely related to uncultivated species. Some archaeal rDNA clones obtained from mine service water and dolomite aquifer water samples were most closely related to environmental rDNA clones from surface soil (soil clones) and marine environments (marine group I [MGI]). Other clones exhibited intermediate phylogenetic affiliation between soil clones and MGI in the Crenarchaeota. Fissure water samples, derived from active or dormant geothermal environments, yielded archaeal sequences that exhibited novel phylogeny, including a novel lineage of Euryarchaeota. These results suggest that deep South African gold mines harbor novel archaeal communities distinct from those observed in other environments. Based on the phylogenetic analysis of archaeal strains and rDNA clones, including the newly discovered archaeal rDNA clones, the evolutionary relationship and the phylogenetic organization of the domain Archaea are reevaluated.     

贵州泥堡卡林型金矿区与非金矿区苔藓植物比较研究

首次报道了贵州普安县泥堡村韭菜烂滩的卡林型金矿区和非金矿区的苔藓。记录了泥堡韭菜烂滩卡林型金矿生苔藓3科8属15种,非金矿生苔藓9科15属20种。通过比较我们得到韭菜烂滩卡林型金矿区和泥堡非金矿区苔藓的相似性系数为11.4%。这表明这两个生境下的苔藓组成差异极大。在这个地区有13种苔藓植物(包括异芽丝瓜藓Pohlia leucostoma、卵蒴丝瓜藓P.proligera、长蒴藓Trematodon longicollis等)只生长在卡林型金矿上,这表明有一些苔藓植物适应卡林型金矿这种基质。也许,在泥堡地区某些苔藓植物的分布与卡林型金矿存在一定的关系。     

Environmental impacts of gold mining in Essakane site of Burkina Faso

In this study, environmental impacts on air, water, and soil pollution caused by the exploit of Essakane gold mine, which is located in North Eastern part of Burkina Faso were investigated. Analyses on drinking water were made in the laboratory to determine the concentration of essential chemicals used in gold mining. Dust fallouts have been measured to assess the level of air pollution. The results showed that exploiting of gold mine directly or indirectly contributes to air pollution in Essakane district. The use of chemicals such as cyanide (industrial gold mining) and mercury (artisanal gold mining) to obtain the gold from the ore constitutes a potential risk for the ecosystem, the local population's health, and livestock production. The results also showed that there is a significant degradation of natural landscape and topography of the soil by open-pits mining (industrial mine) and holes dug by artisanal miners.     

Utilization of nitriles by yeasts isolated from a Brazilian gold mine

Yeast strains from the genera Candida, Debaryomyces, Aureobasidium, Geotrichum, Pichia, Rhodotorula, Tremella, Hanseniaspora, and Cryptococcus were isolated from samples of a gold mine from liquid extraction circuit. These strains were tested for their ability to utilize acetonitrile at 12 mM as the sole nitrogen source. The yeasts that grew using acetonitrile at 12 mM were tested in the presence of acetonitrile, isobutyronitrile, methacrylnitrile, and propionitrile at concentrations of 12, 24, 48, 97, and 120 mM. One strain was selected for each nitrile and the concentration of nitrile in which the best growth occurred. Cryptococcus sp. strain UFMG-Y28 had a better growth on 120 mM propionitrile and 97 mM acetonitrile, Rhodotorula glutinis strain UFMG-Y5 on 48 mM methacrylnitrile, and Cryptococcus flavus strain UFMG-Y61 on 120 mM isobutyronitrile. The utilization of different nitriles and amides by yeast strains involves hydrolysis in a two-step reaction mediated by both inducible and intracellular nitrile hydratase and amidase.     

Chromosomal aberration assessment of silver rasbora fish (Rasbora tornieri) living near gold mine area with heavy metal contamination

This study aimed to investigate arsenic (As), cadmium (Cd), chromium (Cr), and lead (Pb), contamination in water, sediment, and Rasbora tornieri samples and chromosomal aberration in R. tornieri near gold mine area compared to unaffected area. The heavy metal concentrations in the samples were analyzed by inductively coupled plasma atomic emission spectroscopy (ICP-OES). The difference of As, Cr, and Pb concentrations in R. tornieri samples between the gold mine and unaffected areas was statistically significant (p < 0.05), but the difference was not significant for Cd. The diploid chromosome number of R. tornieri from both areas was 2n = 50, and the chromosomal aberrations of R. tornieri were higher in the gold mine area than those in the unaffected area. There are six types of chromosomal aberrations, including centric fragmentation (CF), centric gap, single chromatid gap, fragmentation, deletion, and polyploidy. The most common chromosomal aberration in the samples from the gold mine area was CF, and the difference in chromosomal aberration and the cells with chromosomal aberrations in R. tornieri between the areas was statistically significant (p < 0.05). The percentages of chromosomal aberrations in the R. tornieri samples from the gold mine and the unaffected areas were 16.60 and 1.66, respectively.     

Identification of sulfate-reducing bacteria in methylmercury-contaminated mine tailings by analysis of SSU rRNA genes

Sulfate-reducing bacteria (SRB) are often used in bioremediation of acid mine drainage because microbial sulfate reduction increases pH and produces sulfide that binds with metals. Mercury methylation has also been linked with sulfate reduction. Previous geochemical analysis indicated the occurrence of sulfate reduction in mine tailings, but no molecular characterization of the mine tailings-associated microbial community has determined which SRB are present. This study characterizes the bacterial communities of two geochemically contrasting, high-methylmercury mine tailing environments, with emphasis on SRB, by analyzing small subunit (SSU) rRNA genes present in the tailings sediments and in enrichment cultures inoculated with tailings. Novel Deltaproteobacteria and Firmicutes -related sequences were detected in both the pH-neutral gold mine tailings and the acidic high-sulfide base-metal tailings. At the subphylum level, the SRB communities differed between sites, suggesting that the community structure was dependent on local geochemistry. Clones obtained from the gold tailings and enrichment cultures were more similar to previously cultured isolates whereas clones from acidic tailings were more closely related to uncultured lineages identified from other acidic sediments worldwide. This study provides new insights into the novelty and diversity of bacteria colonizing mine tailings, and identifies specific organisms that warrant further investigation with regard to their roles in mercury methylation and sulfur cycling in these environments.