不同沉积环境中几种厌氧细菌的组成与分布

在严格的厌氧条件下,用MPN计数的方法测定了几种不同沉积环境的地质样品中的硫酸盐还原细菌、发酵性细菌、产甲烷细菌的数量;观察了不同沉积类型的样品中产甲烷细菌的种类和代谢类型;比较了岩芯等典型样品的产甲烷能力。结果表明,陆相沉积物中的硫酸盐还原细菌的分布,受沉积深度的限制,其数量与沉积深度之间有较好的负相关性。在同一沉积环境的相同层位中,发酵性细菌以及厌氧纤维素分解细菌的数量随有机质或纤维素的含量而变化。在各种沉积环境中,产甲烷细菌的数量都明显少于其他非产甲烷细菌。用MPN计数的方法通常检测不到岩芯样品中的产甲烷细菌。现代海相表层沉积物的样品产甲烷细菌主要是甲烷杆菌属(Methanobactertum),其营养类型为H2／CO2。现代陆相沉积物和成岩早期的样品,产甲烷细菌的类群比较复杂,营养类型为H2／CO2和乙酸。成岩中晚期的样品中,产甲烷细菌以甲烷球菌属(Methanococcus)为主,营养类型为H 2／CO2．     

沼气池中产氢细菌的研究

1. 在沼气发酵污泥的富集培养物中加入薯芋粉可以旺盛地产氢。这是富集培养沼气发酵污泥中的产氢细菌的较好方法。 2. 我们采用这一方法从沼气池中分离出24株产氢细菌,其产氢置因菌株的种类和发酵基质的不同而异。根据它们的分类特征分别属肠杆菌科(Enterbacteriace)和芽孢杆菌科(Bacil-laceae)。肠杆菌科中有五个种：阴沟肠杆菌(Entcrbacter cloacae),大肠埃希氏菌(Escheriehiacoli),粘质沙雷氏菌(Serratia marcescens),弗氏柠檬酸杆菌(Citrobacter freundii)和蜂房哈夫尼菌(Hafnia alvei)(暂定)。芽孢杆菌科中仅有一个种,即丙酮丁醇梭菌(Clostridium acetobutyltcum)。 3. 各类菌的相对数量以肠秆菌占优势,占总数的58.3％。其次是粘质沙雷氏菌,占16.7％。丙酮丁酵梭菌占12．5％。其他各菌数量较少。 4. 将这些产氢菌与甲烷菌的富集培养物进行混合培养,可大大提高甲烷产量,而二氧化碳显著降低,以至检测不到。     

PCR-SSCP技术分析碱度影响下硫酸盐还原反应器中微生物群落动态

采用PCR-单链构象多态性(SSCP)技术和杂交技术, 分析进水碱度降低引起的产酸-硫酸盐还原反应器中微生物群落动态. 试验结果表明, 碱度人为控制在4000 mg/L, 硫酸盐负荷率为4.8 kg/(m³·d)时, 反应器25天启动成功, 硫酸盐去除率达87%, 此时Lactococcus sp., Anaerofilum sp.和Kluyvera sp.等占优势. 当短时间降低进水碱度至1000 mg/L以下时, 硫酸盐去除率迅速降到35%; 继而提高并稳定进水碱度为3000 mg/L时, 硫酸盐去除率稳定在55%, Dysgonomonas sp., Sporobacte sp., Obesumbacterium sp.和Clostridium sp.得以富集, 同Lactococcus sp., Anaerofilum sp.和Kluyvera sp.一起构成优势菌群. 碱度继续降低并稳定至1500 mg/L时, 硫酸盐的去除率上升并稳定在70%, Dysgonomonas sp., Sporobacter sp.和Obesumbacterium sp.消亡, 而Desulfovibrio sp.和Clostridium sp.的某些菌种得以大量富集. 为探讨系统所能承受的最低碱度阈值, 再次将碱度降低至400 mg/L, 发现硫酸盐去除率、pH值和出水碱度迅速下降, Petrotoga sp., Prevotella sp., Kluyvera sp.和Neisseria sp.的数量水平明显降低, 杂交显示硫酸盐还原菌(SRB)的数量也随之降低. 微生物群落结构特征和多样性分析表明, 种泥中的SRB种群异常丰富, 碱度降低使种群的丰度逐渐趋于单一. 常驻种群绝大多数为发酵产酸菌(FAB), 其中Firmicute门所占的比例最大, 而SRB所占数量比例极少. 这种群落结构体现了FAB与SRB之间的协同代谢作用, 从而维持着系统硫酸盐较高去除率和运行稳定性.     

食品抗氧剂-D-异抗坏血酸的研究——Ⅰ.2-酮基-D-葡萄糖酸产生菌的筛

从199株细菌中筛选出产2-酮基-D-葡萄糖酸的高产菌株2株。产物对葡萄糖的克分子转化率达85.93%、87.56%以上。经生物学鉴定,菌株E301为恶臭假单胞菌(Pseudomonas putida),菌株E54为产碱菌属的一个新种,为产酮产碱菌(Alcaligenes ketogenes nov.sp.)。将发酵产物及其转化成的D-异抗坏血酸钠样品经红外吸收光谱比较,结果均与标准品相同。可确定这两株菌的发酵产物确系2-酮基-D-葡萄糖酸钙。     

海水养殖场沉积物中硝酸盐还原菌种群分析

通过对福建省沿海海水养殖场沉积物中参与氮循环的各生理群细菌数量分析 ,发现氨化和硝酸盐还原细菌是优势生理菌群 ,同时 ,表层泥样中的硝酸盐还原菌数量明显高于深层泥样。从该环境中分离获得 106株细菌 ,其中 58株具有硝酸盐还原能力 ,初步鉴定表明它们主要为芽孢杆菌属 (Bacillus)、盐芽孢杆菌属 (Halobacillus)、短芽孢杆菌属 (Brevibacil lus)、动性球菌属 (Planococcus)和动性杆菌属 (Plano     

利用氢和二氧化碳生长的一个泥杆菌新种——嗜氢泥杆菌

从降解有机物产生甲烷的富集培养物中分离到一株利用巴豆酸的严格厌氧革兰氏阴性杆菌.此菌利用氢和二氧化碳或甲酸合成乙酸,发酵巴豆酸产生丁酸和乙酸,发酵葡萄糖、果糖、麦芽糖和半乳糖产生乙酸、乙醇和氢或甲酸,不利用延胡索酸、酒石酸和柠檬酸生长.该菌不产生芽孢,不还原硫酸盐,触酶反应阴性,DNA的G+C含量为30.7±1mol%,是泥杆菌属中的一个新种,定名为嗜氢泥杆菌(Ilyobacter hydrogenotrophicus nov.sp.).     

脱氮硫杆菌生长特性及其对SRB生长的影响*

由土壤中分离得到一株自养型的脱氮硫杆菌 (Thiobacillusdenitrificans,硫杆菌属 ,硫杆菌科 ,革兰氏阴性化能自养细菌 ) ,该菌株的最佳生长 pH为 7 0。将此菌株与硫酸盐还原菌 (SulfateReducingBacteria ,SRB ,脱硫弧菌属 ,革兰氏阴性厌氧细菌 )混合培养 ,测定SRB的菌量变化 ,结果表明 ,脱氮硫杆菌的生长抑制了硫酸盐还原菌的生长 ,降低了SRB的腐蚀性的代谢产物硫化物的浓度 ,腐蚀速率降低 ,有利于防治SRB引起的微生物腐蚀。     

滨海湿地甲烷产生途径和产甲烷菌研究进展

滨海湿地在全球碳循环中起着重要的作用,其甲烷排放量占全球海洋甲烷排放的75%.本文综述了滨海湿地主要甲烷产生途径、产甲烷菌种类及其影响因子.滨海湿地SO₄^2-含量丰富,乙酸发酵和H₂/CO₂途径产甲烷受抑制,乙酸营养型和氢营养型产甲烷菌丰度较低;而利用甲胺类等“非竞争性”底物的C1甲基化合物歧化途径不受硫酸还原菌竞争底物的限制,兼性营养型产甲烷菌成为产甲烷优势菌.盐度与SO₄^2-含量和植被类型密切相关,影响竞争性电子和产甲烷底物的种类和含量,对甲烷产生途径和产甲烷菌群落结构有重要影响.目前,滨海湿地产甲烷菌群落结构、甲烷产生途径的关键控制因素尚需明确,其对甲烷排放的影响有待进一步研究.     

甲烷氧化细菌的一个新种

从沼气发酵装置中,分离出一株H型专性甲烷氧化细菌81Z菌株。它具有甲烷氧化细菌的一般典型性状。根据其极生单鞭毛和过氧化氢酶阴性的特征,该菌株明显地区别于已知的任何一种甲烷氧化细菌,被认为是一个新种并命名为沼气甲基弯菌(Methylosinus methanica)。本文还讨论了它在厌氧条件下的生命活动。     

莫克兰冷泉区沉积物原核微生物群落组成及其环境响应

【目的】冷泉系统广泛存在于大陆边缘地区,其典型特征是在海底渗漏出大量富含以甲烷为主的碳氢化合物和硫化氢等成分的低温流体。冷泉也因其独特的地球化学条件孕育着独特的原核微生物群落结构,然而,原核微生物组成与冷泉环境之间的响应关系却并不清楚。【方法】本文以莫克兰大陆边缘活跃冷泉区沉积物柱状样为研究对象,沿深度剖面分析了沉积物中的CH4以及孔隙水SO42–、H2S浓度等关键地球化学参数,并基于16S rRNA基因高通量测序对冷泉沉积物原核微生物的群落结构及其空间变化进行了系统分析。【结果】根据其硫酸盐-甲烷浓度剖面特征,从上向下,将沉积物垂向剖面划分为硫酸盐还原区(SZ)、硫酸盐-甲烷转换区(SMTZ)和产甲烷区(MZ)。通过原核微生物α多样性与基因定量研究发现,随着深度增加微生物多样性与丰度呈逐渐降低的趋势。16SrRNA基因高通量测序结果表明,SZ中以硫氧化细菌γ-变形菌纲、α-变形菌纲和埃普西隆杆菌门为主,且以硫酸盐为电子受体的与有机质降解相关的原核微生物JS1、绿弯菌门、洛基古菌纲、深古菌纲及底栖古菌纲的相对含量也较高;SMTZ存在较高含量的ANME-1a、ANME-1b与SEEP-S...     

Anaerobic Phenol Degradation by Microorganisms of Swine Manure

Swine manure contains diverse groups of aerobic and anaerobic bacteria. An anaerobic bacterial consortium containing sulfate-reducing bacteria (SRB) and acetate-utilizing methanogenic bacteria was isolated from swine manure. This consortium used phenol as its sole source of carbon and converted it to methane and CO₂. The sulfate-reducing bacterial members of the consortium are the incomplete oxidizers, unable to carry out the terminal oxidation of organic substrates, leaving acetic acid as the end product. The methanogenic bacteria of the consortium converted the acetic acid to methane. When a methanogen inhibitor was used in the culture medium, phenol was converted to acetic acid by the SRB, but the acetic acid did not undergo further metabolism. On the other hand, when the growth of SRB in the consortium was suppressed with a specific SRB inhibitor, namely, molybdenum tetroxide, the phenol was not degraded. Thus, the metabolic activities of both the sulfate-reducing bacteria and the methanogenic bacteria were essential for complete degradation of phenol. Received: 31 January 1997 / Accepted: 7 March 1997     

Competition for hydrogen between sulphate-reducing bacteria and methanogenic bacteria from the human large intestine

Sulphate-reducing activity in human faecal slurries was followed by measuring sulphide production. Sulphate-reducing bacteria (SRB) were found to outcompete methanogenic bacteria (MB) for the mutual substrate hydrogen in faecal slurries from methane- and non-methane-producing individuals mixed together. When molybdate (20mmol/l) was added to these slurries, sulphate reduction was inhibited and methanogenesis became the major route of electron disposal. Sulphide production was stimulated by the addition of 20 mmol/1 sulphate in non-methanogenic but not in methanogenic slurries. In methanogenic slurries that contained the methanogen inhibitor 2-bromoethanesulphonic acid (BES), hydrogen accumulated whilst sulphide levels were unaffected, confirming the absence of SRB in methanogenic faeces. The addition of nitrate (10 mmol/l) to faecal slurries completely inhibited methanogenesis but only slightly reduced sulphate reduction. The sulphated mucopolysaccharides, chondroitin sulphate and mucin, strongly stimulated sulphide production in non-methanogenic faecal slurries only, suggesting that these substances may be a potential source of sulphate in the large gut.     

Competition for hydrogen between sulphate-reducing bacteria and methanogenic bacteria from the human large intestine

Sulphate-reducing activity in human faecal slurries was followed by measuring sulphide production. Sulphate-reducing bacteria (SRB) were found to outcompete methanogenic bacteria (MB) for the mutual substrate hydrogen in faecal slurries from methane- and non-methane-producing individuals mixed together. When molybdate (20 mmol/l) was added to these slurries, sulphate reduction was inhibited and methanogenesis became the major route of electron disposal. Sulphide production was stimulated by the addition of 20 mmol/l sulphate in non-methanogenic but not in methanogenic slurries. In methanogenic slurries that contained the methanogen inhibitor 2-bromoethanesulphonic acid (BES), hydrogen accumulated whilst sulphide levels were unaffected, confirming the absence of SRB in methanogenic faeces. The addition of nitrate (10 mmol/l) to faecal slurries completely inhibited methanogenesis but only slightly reduced sulphate reduction. The sulphated mucopolysaccharides, chondroitin sulphate and mucin, strongly stimulated sulphide production in non-methanogenic faecal slurries only, suggesting that these substances may be a potential source of sulphate in the large gut.     

Application of the fluorescent-antibody technique to the study of a methanogenic bacterium in lake sediments.

Fluorescent antibody (FA) was prepared for a methanogenic bacterium isolated from Wintergreen Lake pelagic sediment. The isolate resembles Methanobacterium formicicum. The FA did not cross-react with 9 other methanogens, including M. formicicum strains, or 24 heterotrophs, 18 of which had been isolated from Wintergreen Lake sediment. FA-reacting methanogens were detected in heat-fixed smears of several different lake sediments and anaerobic sewage sludge. Pretreatment of all samples with either rhodamine-conjugated geletin or bovine serum albumin adequately controlled nonspecific absorption of the FA. Autofluorescent particles were observed in the sediment samples but, with experience, they could easily be distinguished from FA-reacting bacteria. FA direct counts of the specific methanogen in Wintergreen Lake sediments were made on four different sampling dates and compared with five-tube most-probable-number estimates of the total methanogenic population that was present in the same samples. The FA counts ranged from 3.1 X 10(6) to 1.4 X 10(7)/g of dry sediment. The highest most-probable-number estimates were at least an order ofmagnitude lower.     

Growth and Activities of Sulfate-Reducing and Methanogenic Bacteria in Human Oral Cavity

Viable counts and activities of sulfate-reducing bacteria (SRB) and methanogenic bacteria were determined in the oral cavities of eight volunteers. Of these, seven harbored viable SRB populations, and six harbored viable methanogenic bacterial populations. Two volunteers classified as type III periodontal patients had both SRB and methanogenic bacteria. Six separate sites were sampled: posterior tongue, anterior tongue, mid-buccal mucosa, vestibular mucosa, supragingival plaque, and subgingival plaque. The SRB was found in all areas in one volunteer, and it was mostly present in posterior tongue, anterior tongue, supragingival, and subgingival plaques in many volunteers. The methanogenic bacteria were mostly found in supragingival and subgingival plaques. The activities of sulfate reduction and methane production were determined in randomly selected isolates. Received: 27 July 2002 / Accepted: 27 August 2002     

Application of the fluorescent-antibody technique to the study of a methanogenic bacterium in lake sediments.

Fluorescent antibody (FA) was prepared for a methanogenic bacterium isolated from Wintergreen Lake pelagic sediment. The isolate resembles Methanobacterium formicicum. The FA did not cross-react with 9 other methanogens, including M. formicicum strains, or 24 heterotrophs, 18 of which had been isolated from Wintergreen Lake sediment. FA-reacting methanogens were detected in heat-fixed smears of several different lake sediments and anaerobic sewage sludge. Pretreatment of all samples with either rhodamine-conjugated geletin or bovine serum albumin adequately controlled nonspecific absorption of the FA. Autofluorescent particles were observed in the sediment samples but, with experience, they could easily be distinguished from FA-reacting bacteria. FA direct counts of the specific methanogen in Wintergreen Lake sediments were made on four different sampling dates and compared with five-tube most-probable-number estimates of the total methanogenic population that was present in the same samples. The FA counts ranged from 3.1 X 10(6) to 1.4 X 10(7)/g of dry sediment. The highest most-probable-number estimates were at least an order ofmagnitude lower.     

Mercury Methylation by the Methanogen Methanospirillum hungatei

Methylmercury (MeHg), a neurotoxic substance that accumulates in aquatic food chains and poses a risk to human health, is synthesized by anaerobic microorganisms in the environment. To date, mercury (Hg) methylation has been attributed to sulfate- and iron-reducing bacteria (SRB and IRB, respectively). Here we report that a methanogen, Methanospirillum hungatei JF-1, methylated Hg in a sulfide-free medium at comparable rates, but with higher yields, than those observed for some SRB and IRB. Phylogenetic analyses showed that the concatenated orthologs of the Hg methylation proteins HgcA and HgcB from M. hungatei are closely related to those from known SRB and IRB methylators and that they cluster together with proteins from eight other methanogens, suggesting that these methanogens may also methylate Hg. Because all nine methanogens with HgcA and HgcB orthologs belong to the class Methanomicrobia, constituting the late-evolving methanogenic lineage, methanogenic Hg methylation could not be considered an ancient metabolic trait. Our results identify methanogens as a new guild of Hg-methylating microbes with a potentially important role in mineral-poor (sulfate- and iron-limited) anoxic freshwater environments.     

Methylotrophic methanogenesis governs the biogenic coal bed methane formation in Eastern Ordos Basin, China

To identify the methanogenic pathways present in a deep coal bed methane (CBM) reservoir associated with Eastern Ordos Basin in China, a series of geochemical and microbiological studies was performed using gas and water samples produced from the Liulin CBM reservoir. The composition and stable isotopic ratios of CBM implied a mixed biogenic and thermogenic origin of the methane. Archaeal 16S rRNA gene analysis revealed the dominance of the methylotrophic methanogen Methanolobus in the water produced. The high potential of methane production by methylotrophic methanogens was found in the enrichments using the water samples amended with methanol and incubated at 25 and 35?°C. Methylotrophic methanogens were the dominant archaea in both enrichments as shown by polymerase chain reaction (PCR)–denaturing gradient gel electrophoresis (DGGE). Bacterial 16S rRNA gene analysis revealed that fermentative, sulfate-reducing, and nitrate-reducing bacteria inhabiting the water produced were a factor in coal biodegradation to fuel methanogens. These results suggested that past and ongoing biodegradation of coal by methylotrophic methanogens and syntrophic bacteria, as well as thermogenic CBM production, contributed to the Liulin CBM reserves associated with the Eastern Ordos Basin.     

Molecular profiling and identification of methanogenic archaeal species from rabbit caecum

During a comparison of 16S rDNA PCR-denaturant gradient gel electrophoresis (DGGE) profiles of methanogenic archaea from rumen fluid, rabbit caecum and pig feces, a unique band common to all rabbit caecum samples was observed. DGGE profiling also showed that the methanogen community from the New Zealand White adult rabbits is different and less complex than the methanogen communities from the rumen and pig feces. Small subunit ribosomal gene sequences of methanogenic archaea were subsequently retrieved from the constructed rabbit caecum 16S rDNA gene library. Results of the phylogenetic analysis indicated that rabbit caecum is inhabited by members of the genus Methanobrevibacter and is possibly one-species dominated, because all the retrieved sequences exhibited similarity values of 99% or higher. This species may well be a novel species of the genus Methanobrevibacter. It belongs to a distinct phylogenetic group containing Methanobrevibacter woesei, Methanobrevibacter thaueri and Methanobrevibacter gottschalkii strains isolated from animal feces, and Methanobrevibacter smithii from the predominating methanogen population of the human large bowel.     

Toxicity of nitrite toward mesophilic and thermophilic sulphate-reducing, methanogenic and syntrophic populations in anaerobic sludge

The various problems associated with treating sulphate-containing wastewaters stem inherently from successful competitive interactions between sulphate reducing bacteria (SRB) and other bacteria involved in the process, resulting in the formation of H₂S. Prevention of in-reactor sulphide generation by use of specific SRB inhibitors presents a potential solution. Nitrite has been reported to be a specific inhibitor of SRB but its possible toxicity to syntrophic and methanogenic members of the anaerobic consortium has not been investigated. In batch activity and toxicity tests, under both mesophilic and thermophilic conditions, nitrite, at concentrations of up to 150 mg L^–1, was found to be ineffective as a specific inhibitor of SRB, and was also shown to have an inhibitory effect on the activity of syntrophic and methane-producing bacteria in mesophilic and thermophilic digester sludge samples.