西太平洋深海海水中乙醛降解菌多样性及群落结构研究

As a major aldehyde pollutant widely existing in industry and our daily life, acetaldehyde is more and more harmful to human health. As characteristic habitat niche, bacteria from deep sea environments are abundant and distinctive in heredity, physiology and ecological functions. Thus, the development of acetaldehyde-degrading bacteria from deep sea provides a new method to harness acetaldehyde pollutant. Firstly, in this study,acetaldehyde-degrading bacteria in the deep sea water of the West Pacific Ocean were enriched in situ and in the laboratory respectively, and then the diversity of uncultured bacteria was studied by using 16 S r RNA genes. Then acetaldehyde-degrading strains were isolated from two samples, including enrichment in situ and enrichment in laboratory samples of deep sea water from the West Pacific Ocean using acetaldehyde as the sole carbon source,and then the ability of acetaldehyde degradation was detected. Our results showed that the main uncultured bacteria of two samples with different enrichment approaches were similar, including Proteobacteria,Actinobacteria, Firmicutes, Cyanobacteria, but the structure of bacterial community were significant different.Four subgroups, α, γ, δ and ε, were found in Proteobacteria group. The γ-Proteobacteria was dominant(63.5%clones in laboratory enriched sample, 75% clones in situ enriched sample). The species belonged to γ-Proteobacteria and their proportion was nearly identical between the two enrichment samples, and Vibrio was the predominant genus(45% in laboratory enriched sample, 48.5% in situ enriched sample), followed by Halomonas(9% in situ enriched sample) and Streptococcus(6% in laboratory enriched sample). A total of 12 acetaldehyde-degrading strains were isolated from the two samples, which belonged to Vibrio, Halomonas,Pseudoalteromonas, Pseudomonas and Bacillus of γ-Proteobacteria. Strains ACH-L-5, ACH-L-8 and ACH-S-12,belonging to Vibrio and Halomonas, have strong ability of acetaldehyde degradation, which could tolerate 1.5 g/L acetaldehyde and degrade 350 mg/L acetaldehyde within 24 hours. Our results indicated that bacteria of γ-Proteobacteria may play an important role in carbon cycle of deep sea environments, especial the bacteria belonging to Vibrio and Halomonas and these strains was suggested for their potentials in government of aldehyde pollutants.     

刺参肠道微生物组成分析及产酶、溶血性试验

对野生和人工养殖刺参的肠壁及内容物中的菌群数量、种类组成进行了研究;并结合产酶试验和溶血性试验,对刺参肠道益生菌做了初步的体外筛选。结果表明,野生刺参肠壁及内容物中的细菌数量分别为(3.30±0.41)×107 cfu/g、(6.39±0.32)×107 cfu/g,养殖刺参肠壁及内容物中的细菌数量分别为(2.83±0.31)×107 cfu/g、(5.67±0.53)×107 cfu/g。野生刺参肠道优势菌为弧菌属(Vibrio),次优势菌为假单胞菌属(Pseudomonas)和希瓦氏菌属(Shewanella);养殖刺参肠道优势菌为弧菌属(Vibrio),次优势菌为假单胞菌属(Pseudomonas)。在224株细菌中,共有160株细菌具有产酶能力,所占比例为71.43%,其中具产蛋白酶、淀粉酶、脂肪酶能力菌株分别为114株、114株、108株,所占比例分别为50.89%、50.89%、48.21%。99株细菌中有23株具有溶血性,所占比例为23.23%。综合分析实验数据,确定6株细菌作为刺参肠道潜在益生菌,菌株代号分别为HS1(Pseudomonas)、HS5(Bacillus)、HS7(Shewanella)、HS8(Vibrio)、HS10(Vibrio)、HS11(Vibrio)。     

海南万宁小海近岸表层海水弧菌组成及其变化

于2006年冬至2007年秋分四个季度对海南万宁小海近岸表层海水的弧菌种类分布及数量变化进行了调查.共分离到27种弧菌属细菌,其中副溶血弧菌、创伤弧菌、溶藻弧菌、霍乱弧菌、河流弧菌和拟态弧菌6种弧菌为主要弧菌.在6种主要弧菌中,溶藻弧菌、副溶血弧菌和霍乱弧菌在夏季和秋季检出率较高;河流弧菌在春季和夏季的检出率较高;拟态...     

南极低温降解菌的筛选、鉴定与低温降解适应性分析

以南极海洋细菌为研究对象,采用2216E和MMC液体培养基结合的方法,从菌种库里筛选南极低温降解菌,进行石油烃降解率测定、表型特征确定和16SrRNA分子鉴定,构建系统发育树,并对南极低温降解菌NJ49的低温降解适应性进行了初步研究。结果表明,3株南极海洋细菌NJ41、NJ49和NJ289可以在以柴油为惟一碳源和能源的培养基中生长,被视为南极低温降解菌;16SrRNA分子鉴定结果表明,南极低温降解菌NJ41、NJ49和NJ289分别属于Planococcus、Shewanella和Pseudoalteromonas属;NJ49通过改变脂肪酸组分的膜修饰方式适应低温降解。     

Seston available as a food resource for the ribbed mussel (Geukensia demissa) in a North American,mid-Atlantic saltmarsh

We studied the composition of the <25 μm seston size fraction as a food resource potentially available to suspension feeding ribbed mussels, Geukensia demissa, over an annual cycle in Canary Creek saltmarsh, Delaware Bay. There were significant seasonal variations in the concentration of particulate organic carbon (POC), particulate organic nitrogen (PON), and total carbohydrate, but not cellulose. The concentration of cellulose, measured by hydrolytic cellulase enzyme assay, was relatively low (seasonal range 24 to 35 μg l⁻¹) and only comprised from 3% of total carbohydrate in May 1996 to 13% in November 1995. We used the biomass of microalgae, estimated from chlorophyll a, and abundance of free-living bacteria and heterotrophic nanoflagellates to calculate each component's equivalent carbon content. Microalgae were the most dominant carbon source (62% annually) among the four identified components (phytoplankton, bacteria, heterotrophic nanoflagellates, and cellulose) in all seasons except in August 1995 when carbon from bacteria was most abundant (55%). The annual average carbon equivalents of heterotrophic nanoflagellates and cellulose were relatively small (2 and 4%, respectively). The total concentration of POC in the seston was much greater than the carbon derived from the four identified components. The proportions that these identified components contributed to POC varied seasonally and combined only accounted for 8–24% of POC. Based on these estimates, the bulk of the POC in Canary Creek marsh was not associated with any of the four components we identified. We suggest that this uncharacterized material was some type of non-lignocellulosic, amorphous detritus of unknown utility as a food resource for ribbed mussels.     

海绵共栖细菌抗菌活性及其群体感应信号的研究

利用平板涂布法从南海海绵生物样品中分离海绵共栖细菌,以枯草芽孢杆菌、金黄色葡萄球菌、大肠杆菌及酿酒酵母作为实验用指示菌对海绵共栖细菌进行了抗菌活性筛选。利用气相质谱联用仪(GC-MS)建立了群体感应信号分子酰基高丝氨酸内酯类化合物(AHLs)的检测方法,分析了活性菌株粗提液中信号分子的产生及种类。同时采用16S rDNA同源性和系统发育分析对活性菌株进行种属鉴定和系统发生学研究。结果表明,在分离到的94株海绵共栖细菌中18株(占19.1%)细菌具有抗菌活性。通过GC-MS检测确定大多数活性菌株都含有AHLs,因此提出了海绵共栖细菌抗菌活性与群体感应之间可能存在必然联系的推测。通过细菌分类鉴定,结果显示具有抗菌活性的细菌大部分属于芽孢杆菌属(Bacillus)。     

海绵共栖细菌抗菌活性及其群体感应信号的研究

利用平板涂布法从南海海绵生物样品中分离海绵共栖细菌,以枯草芽孢杆菌、金黄色葡萄球菌、大肠杆菌及酿酒酵母作为实验用指示菌对海绵共栖细菌进行了抗菌活性筛选。利用气相质谱联用仪(GC-MS)建立了群体感应信号分子酰基高丝氨酸内酯类化合物(AHLs)的检测方法,分析了活性菌株粗提液中信号分子的产生及种类。同时采用16S rDNA同源性和系统发育分析对活性菌株进行种属鉴定和系统发生学研究。结果表明,在分离到的94株海绵共栖细菌中18株(占19.1%)细菌具有抗菌活性。通过GC-MS检测确定大多数活性菌株都含有AHLs,因此提出了海绵共栖细菌抗菌活性与群体感应之间可能存在必然联系的推测。通过细菌分类鉴定,结果显示具有抗菌活性的细菌大部分属于芽孢杆菌属(Bacillus)。     

利用凝胶双向扩散试验研究海洋发光细菌的分类关系

细菌分类,向以形态学和生理生化特征为主要依据。近年来,由于分子生物学的进展,提供了很多新技术,使细菌分类的研究进入分子水平。此类研究结果反映了细菌之间的亲缘关系,因而更确切地反映了细菌各属、种之间的进化联系。凝胶双向扩散试验属于免疫学方     

条斑紫菜外生细菌的遗传多样性

为研究藻际微生物多样性与紫菜健康养殖的关系,从江苏南通(养殖)、山东青岛(野生)、浙江温州(养殖)三地的条斑紫菜叶状体和江苏东台条斑紫菜贝壳丝状体及其周围的海水样品中分离了条斑紫菜外生细菌。根据菌落特征和菌体形态将获得的63株细菌分为21种表型。16SrDNA-V3片段DGGE分析结合序列测序比对结果表明:这些菌株与假交替单孢菌(Pseudoalteromonas)、嗜冷单胞菌(Psychrobacter)和芽孢杆菌(Bacillus)等10属相应菌株具有较高的同源性。通过对条斑紫菜外生细菌区系多样性分析得出:条斑紫菜藻体外生细菌组成与生活环境相关,但不同于周围海水细菌群落;不同来源(地点)、不同生理状况(健康与病烂)和不同生长阶段(叶状体和丝状体)的条斑紫菜外生菌种群组成差异较大。许多外生细菌对常见测试菌、海水养殖动物致病菌和人致病菌具有抑菌活性,其抑菌特点与紫菜生理性状相关。从健康条斑紫菜样品分离的外生菌中假交替单孢菌占优势,病烂紫菜未分离到假交替单孢菌,提示假交替单孢菌可能与紫菜健康生长有密切关系。     

我国近海中度嗜盐菌的分离筛选及其产酶多样性分析

为了解我国近海中度嗜盐菌系统发育多样性及产酶特性,从天津、山东、江苏、福建、海南等地近海采集样品为研究对象,分离筛选得到108株中度嗜盐菌,其中有26株至少产一种酶。通过对其形态特征、生理生化、16SrRNA基因序列及系统进化树进行分析,将26株菌鉴定为细菌域的Halomonas、Idiomarina、Virgibacillus、Pontibacillus、Oceanobacillus、Halobacillus和Marinilactibacillus属的菌株。它们与相应属的模式菌株16SrRNA基因序列相似性在97%~100%不等,其中有些菌株可能代表不同的分类单元。底物特异性试验表明,分离的26株中度嗜盐菌13株产蛋白酶,19株产淀粉酶,13株产酯酶,4株产纤维素酶。其中6株产3种酶,11株产2种酶。研究结果表明中度嗜盐菌具有系统发育和产酶多样性,同时蕴藏着较多新的微生物类群,为嗜盐微生物资源应用开发提供了参考。     

Characterization of bacterial communities associated with seasonal water masses from Tongyoung in South Sea of Korea

Understanding the patterns of microbial diversity and their composition in seawater is necessary to assess the impacts of microbes on marine ecosystem. Although the potential roles of microbes in the ocean have been studied, their diversity and relationship with environmental factors remain unclear. In this study, we adopted a 16S rRNA gene tag-pyrosequencing technique to investigate the bacterial communities associated with two different water masses from Tongyoung in the South Sea of Korea. Our results revealed highly diverse bacterial communities up to 900 Operational Taxonomic Units (OTUs) estimated from each seawater which was collected in the month of March and May, when the environmental conditions including temperature differed significantly: 7.2°C and 17.6°C in March and May, respectively. Altogether, 13 bacterial phyla were recovered from the seawater, of which Proteobacteria was the most dominant group. In addition, the value of the Shannon index, which measures the evenness of the distribution of individuals among OTUs, in May is higher than that in March, indicating that it displays a wider diversity of bacteria. Interestingly, the proportion of pathogenic bacteria was significantly increased in the month of May compared to March, suggesting that pathogenic bacteria were increasingly emerging in May. In particular, Pseudoalteromonas and Vibrio spp. were determined as major pathogenic bacteria from both water masses, of which Vibrio spp. were dominant.     

从养殖贝类和大菱鲆分离的海洋发光弧菌的分类地位分析

海洋发光弧菌是海水养殖中重要的条件致病菌。从福建省晋江市附近水产养殖场花蛤、牡蛎软组织及波罗地海大菱鲆肠道内含物中分离得到16 株发光细菌, 采用ARDRA（amplified ribosomal DNArestriction analysis）、16S rDNA 序列测定、Biolog 碳源代谢分析及药敏试验等方法对...     

A study of crude oil‐degrading bacteria from mangrove forests in the Persian Gulf

Mangroves are coastal ecosystems, found in tropical and subtropical regions around the world. They are found in the transitional zones between land, sea, and rivers. Petroleum hydrocarbons are the most common environmental pollutants, and oil spills pose a great hazard to mangroves forests. This research was focused on the isolation and characterization of crude oil‐degrading bacteria from mangrove ecosystems at the Persian Gulf. Sixty‐one crude oil‐degrading bacteria were isolated from mangrove samples (plant, sediment, and seawater) that enriched in ONR7a medium with crude oil as only carbon source. Some screening tests such as growth at high concentration of crude oil, bioemulsifier production, and surface hydrophobicity were done to select the most efficient strains for crude oil degradation. Molecular identification of strains was carried out by amplification of the 16S rRNA gene by PCR. The results of this study were indicated that the quantity of crude oil‐degrading bacteria was higher in the root of mangrove plants compare to other mangrove samples (sediment and seawater). Also, identification results confirmed that these isolated strains belong to Vibrio sp. strain NW4, Idiomarina sp. strain BW32, Kangiella sp. strain DP40, Marinobacter sp. strain DW44, Halomonas sp. strain BS53, and Vibrio sp. strain DS35. The application of bioremediation strategies with these bacteria can reduce crude oil pollution in this important marine environment.     

Bacterial production,glucosidase activity and particle-associated carbohydrates in Dona Paula bay,west coast of India

Size-fractionated bacterial production, abundance and α- and β- glucosidase enzyme activities were studied with respect to changes in hydrography, total suspended matter (TSM), chlorophyll a, particulate organic carbon and nitrogen ratio (POC:PON), 1.5 M NaCl-soluble and 10 mM EDTA-soluble carbohydrates (Sal-PCHO and CPCHO) and transparent exopolymeric particles (TEP) in the surface waters from July 1999–2000 at a shallow coastal station in Dona Paula Bay, west coast of India. The bulk of the total bacterial production and glucosidase activity were associated with particles (75% and >80%, respectively). Total bacterial production was linearly correlated to chlorophyll a (r = 0.513; p < 0.05) whereas enzyme activity was significantly correlated to TSM (α-glucosidase: r = 0.721 (p < 0.001); β-glucosidase: r = 0.596 (p < 0.01)). Both α-glucosidase (r = 0.514; p < 0.05) and β-glucosidase enzymes (r = 0.598; p < 0.01) appeared to be involved in the degradation of CPCHO and Sal-PCHO, respectively. Changes in α-glucosidase/β-glucosidase ratios highlighted the varying composition of particulate organic matter. The bacterial uptake of ¹⁴C-labeled bacterial extracellular carbohydrate measured over 11 days showed a strong linear correlation between ¹⁴C-uptake and bacterial production using tritiated thymidine. The turnover rate of ¹⁴C-labeled carbohydrate-C was 0.52 d⁻¹, higher than the estimated annual mean potential carbohydrate carbon turnover rate of 0.33 ± 0.2 d⁻¹. Our study suggests that carbohydrates derived from sediments may serve as an important alternative carbon source sustaining the bacterial carbon demand in the surface waters of Dona Paula Bay.     

虾池养殖环境有机污染物降解细菌的筛选

在实验室条件下通过富集培养共分离出 1 2 1株菌进行筛选。利用需氧性、BOD2 / COD0 、胞外酶等指标进行筛选 ,最后筛选到 7株对富营养有机物具有较高降解能力的细菌 ,它们均能产生酪蛋白酶、明胶酶、脂酶 (Tween- 80 )、淀粉酶其中 4株细菌还能产生卵磷脂酶。通过测定 BOD5及 COD来衡量 7株细菌利用对虾饵料的能力 ,5d内对虾饵料培养基的 CODMn去除率达到 59.6%～ 79.2 %。用常规生理生化方法将细菌鉴定到属 ,其中 1株为弧菌属细菌 (Vibrio spp.) ,6株为芽孢杆菌属细菌(Bacillusspp.)。     

Phylogenetic diversity of planktonic bacteria in the Chukchi Borderland region in summer

Planktonic bacteria are abundant in the Chukchi Borderland region. However, little is known about their diversity and the roles of various bacteria in the ocean. Seawater samples were collected from two stations K2S and K4S where sea ice was melting obviously. The analysis of water samples with fluorescence in situ hybridization (FISH) showed that DMSP-degrading bacteria accounted for 13% of the total bacteria at the station K2S. No aerobic anoxygenic phototrophic (AAP) bacteria were detected in both samples. The bacterial communities were characterized by two 16S rRNA gene clone libraries. Sequences fell into four major lineages of the domain Bacteria, including Proteobacteria (Alpha, Beta and Gamma subclasses), Bacteroidetes, Actinobacteria and Firmicutes. No significant difference was found between the two clone libraries. SAR11 and Rhodobacteraceae clades of Alphaproteobacteria and Pseudoalteromonas of Gammapro-teobacteria constituted three dominant fractions in the clone libraries. A total of 191 heterotrophic bacterial strains were isolated and 76% showed extracellular proteolytic activity. Phylogenetic analysis reveals that the isolates fell into Gammaproteobacteria, Bacteroidetes, Actinobacteria and Firmicutes. The most common genus in both the bacterial isolates and protease-producing bacteria was Pseudoalteromonas. UniFrac data showed suggestive differences in bacterial communities between the Chukchi Borderland and the northern Bering Sea.     

胶州湾沉积物中石油降解菌的降解能力及多样性研究

2013年青岛输油管道爆炸,大量石油污染了附近海岸。课题组采集了污染的沉积物样品,以原油为唯一碳源和能源,富集了四个石油降解菌群。生物多样性和群落分析表明,Luteibacter、Parvibaculum 和属于食烷菌科的一个属是降解菌群的主要优势菌,都属于变形菌门。从石油降解菌群中分离筛选,获得了9株具有不同16S rRNA基因序列的降解菌,分别属于8个属。重量法测定降解菌的石油降解率,其中5株的石油降解率大于30%。GC-MS分析结果表明,石油降解菌多倾向于降解烷烃,对多环芳烃的降解能力较差,其中5株细菌的烷烃降解率较大,仅1株菌D2对多环芳烃的降解率较大,其降解率在34.9%到77.5%。通过对高通量数据的分析,表明食烷菌属是菌群A和菌群E的主要降解菌群,其中筛选获得的菌株E4可能是菌群E的一株优势降解菌。本研究所筛选菌株证明了其石油降解潜力,为油污染海滩生物修复提供了菌株资源。     

Continental shelf benthos off Otago Peninsula,New Zealand

Benthic macrofauna of the continental shelf off Otago Peninsula, south-eastern New Zealand (45°51′S, 170°52′E) was surveyed by dredge sampling during 1973–1975. Numerical classification (Canberra metric coefficient and flexible sorting) was used to produce site groups and species groups, and three major benthic communities were recognised: a shallow-water (14–25 m) fauna inhabiting well-sorted fine sand, a mid-shelf fauna (concentrated in the depth range 50–76 m) associated with sediments containing the greatest proportions of gravel and siltclay, and a predominantly sand-bottom fauna occurring mainly on the outer shelf (87–150 m). All station groups were dominated numerically by polychaetes (mean of 36·6–56% of individuals) with Mollusca (13·8–25%) or Crustacea (12·1–19·4%) the next most abundant group.The inshore sand fauna was the most distinct, characteristic elements being the trochid gastropod Antisolarium egenum, an amphipod of the genus Hippomedon and dense patches of the spionid polychaete Spiophanes bombyx. Diagnostic species of the mid-shelf mixed sediments were Lepidonotus jacksoni, Psammolyce antipoda, Lumbrineris brevicirra and Phyllamphicteis foliata (Polychaeta), Terenochiton otagoensis, Micrelenchus caelatus caelatus, Maoricolpus roseus roseus and Zegalerus tenuis (Mollusca), Ampelisca chiltoni (Amphipoda) and Amphipholis squamata (Ophiuroidea). Outer shelf sand stations were faunally less distinct, but among the more characteristic species were Euthalenessa fimbriata, Sigalion sp. and Euchone sp. (Polychaeta) and Gari stangeri (Bivalvia). Several abundant species were widely distributed among station groups, notably Nephtys macroura, Lumbrineris magalhaensis, Phyllochaetopterus socialis and Owenia fusiformis (Polychaeta) and Nucula nitidula and Tawera spissa (Bivalvia).Free-living lunulitiform Bryozoa of the genus Otionella were a characteristic component of inner and outer shelf sand faunas, and their inshore penetration probably marks the outer limit of a turbulent zone.Species diversity (Margalef's d) varied markedly across the shelf and appeared to be related primarily to the degree of sediment heterogeneity. Mixed sediments of the middle shelf were particularly rich in species.     

Hydrocarbon-utilising micro-organisms from Dona Paula Bay,Goa

Twenty-three hydrocarbon-utilising bacteria and one yeast were isolated, using enrichment techniques, from water and sediment samples. Vibrio and Pseudomonas were the predominant genera. Of the different organisms screened, Bacillus, Candida and Arthrobacter sp. exhibited the widest range of hydrocarbon-utilising profiles. Arabian Sea crude and kerosene supported the growth of most of the isolates.     

褐藻酸降解菌在海带绿烂病发生中的作用

从海带幼苗上病烂明显处分离得到 2 3株褐藻酸降解菌 ,并对其菌落形态、细胞形态及革兰氏染色反应等进行了观察。通过对褐藻酸钠的降解实验 ,筛选出 4株具较高降解能力的菌株。感染实验表明 ,分离到的褐藻酸降解菌为海带幼苗病烂的主要病原菌。