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1.
Activity and numbers of heterotrophic bacteria have indicated that, as expected, Prevost Lagoon is more eutrophic than Arcachon Bay. Amplification and sequence analysis of the 16S rRNA genes from DNA samples extracted directly from the environment allow the determination of phylogenetic relationships among members of microbial communities in natural ecosystems without the need for cultivation. Analysis of partial 16S rRNA gene sequences obtained from Stations A and 11 revealed that, in both environments, a relatively large number of clones related to Cytophaga/FlexibacterBacteroides as well as to -Proteobacteria were found. One hundred percent similarity with the sequences of the data bases were not found for any of the more than a hundred clones studied. In fact for most clones maximum similarity was below 95% for the nucleotide series sequenced. Similarity was not higher with any of the sequences found for the 14 isolates (pure cultures) obtained from the same samples. Redundancy, i.e. number of identical sequences, was higher in the samples from Arcachon. In addition, sequences related to representatives of ten major phylogenetic branches of Eubacteria were obtained from Prevost Lagoon, however only five branches were represented by the data from Arcachon. These findings indicate a higher bacterial diversity in Prévost Lagoon.  相似文献   

2.
Sediment samples collected from coastal lagoons on the French Mediterranean (Prévost Lagoon) and Atlantic coasts (Arcachon Bay and Certes fishponds) have been studied in order to determine the population densities and the species diversity of the different groups of anoxygenic phototrophic bacteria (purple sulfur bacteria, purple nonsulfur bacteria and green sulfur bacteria) present in these ecosystems. Several strains of each group were isolated in pure culture and characterized by their physiological properties. The occurrence of purple nonsulfur bacteria in organic rich sediments of the Arcachon Bay and the dominance of purple sulfur bacteria in the Prévost lagoon and Certes fishponds are discussed with respect to their community structure and abundance. The diversity differences of the phototrophic bacterial strains isolated from both environments are also discussed.  相似文献   

3.
Abstract In the shallow lagoon of Prévost (43°30'N, 3°54'E; French Mediterranean coast), red waters occurring periodically during warm summers are a consequence of a succession of ecological events beginning in the early spring with a bloom of algae ( Ulva lactuca ). In summer 1977, a red water was analyzed; in the early summer, the water turned anoxic and became rich in sulfide which originated from sulfate reduction in the first 10 cm of the sediment. Numbers of both phototrophic and sulfate-reducing bacteria (SRB) increased during spring and summer, and the genera in the prevailing populations did not change: Thiocapsa (80%) among the phototrophic bacteria and Desulfovibrio and Desulfobacter among the SRB. They were also dominant during the period of red waters. A few Chromatium and Thiocystis species were also identified. During red water periods, these bacteria grew very actively, removing all the sulfide produced by SRB, and accumulated in the whole water column. Consequently, the sulfate level increased to 5 mmol·1−1 higher than the theoretical sulfate level calculated from salinity, showing the active oxidation of sulfide by phototrophic bacteria. After the dystrophic crisis, oxic conditions were reestablished and the phototrophic bacterial biomass was partly grazed by zoobenthos organisms which densely populated the sediment surface.  相似文献   

4.
Representatives of the genus Legionella were detected by use of a real-time PCR method in all water samples collected directly after treatment from 16 surface water (SW) supplies prior to postdisinfection and from 81 groundwater (GW) supplies. Legionella concentrations ranged from 1.1 x 10(3) to 7.8 x 10(5) cells liter(-1) and were significantly higher in SW treated with multiple barriers at 4 degrees C than in GW treated at 9 to 12 degrees C with aeration and filtration but without chemical disinfection. No Legionellae (<50 CFU liter(-1)) were detected in treated water by the culture method. Legionella was also observed in untreated SW and in untreated aerobic and anaerobic GW. Filtration processes in SW and GW treatment had little effect or increased the Legionella concentration, but ozonation in SW treatment caused about 1-log-unit reduction. A phylogenetic analysis of 16S rRNA gene sequences of 202 clones, obtained from a selection of samples, showed a high similarity (>91%) with Legionella sequences in the GenBank database. A total of 40 (33%) of the 16S rRNA gene sequences obtained from treated water were identified as described Legionella species and types, including L. bozemanii, L. worsleiensis, Legionella-like amoebal pathogen types, L. quateirensis, L. waltersii, and L. pneumophila. 16S rRNA gene sequences with a similarity of below 97% from described species were positioned all over the phylogenetic tree of Legionella. Hence, a large diversity of yet-uncultured Legionellae are common members of the microbial communities in SW and GW treated at water temperatures of below 15 degrees C.  相似文献   

5.
The high biodiversity observed in coastal lagoons can often be related to a very high degree of environmental heterogeneity (β-biodiversity). In this article, I describe many physical factors that contribute to create environmental heterogeneity and different habitats within the Bassin d’Arcachon, a mesotidal lagoon in S.W. France. The microbial biodiversity has been studied in this coastal lagoon by culture-dependent and culture-independent techniques. The sediments of the meadows of the seagrass Zostera noltii have received a specific focus. During the last 15 years, several novel bacterial species have been isolated from these sediments and obtained in culture. Among these, five different species have been fully described as novel species according the Bacteriological Code and two of them represent also the type strain for a novel genus. The bacterial biodiversity has also been studied by molecular ecology techniques. An early study was focused on water column bacteria and a comparison has been made with the same community from the more eutrophic Prévost lagoon (France, Mediterranean coast). A clone library was also constructed for 16-S rRNA genes isolated and amplified from the 1–2 cm depth layer of the Z. noltii sediment. These studies showed a high degree of biodiversity and the importance of sulfate-reducing bacteria in the sediment, pointing to species that have not been obtained in culture. Collectively, these data show that there is still a large discrepancy between culture-dependent and culture-independent prokaryotic biodiversity. It is important to link the functional biogeochemical approach with the biodiversity approach. For the Bassin d’Arcachon studies, the biogeochemical approach has successfully nurtured the biodiversity studies, while the reverse has not yet been the case as the biogeochemical studies have lumped prokaryotes in functional groups. Guest editors: A. Razinkovas, Z. R. Gasiūnaitė, J. M. Zaldivar & P. Viaroli European Lagoons and their Watersheds: Function and Biodiversity  相似文献   

6.
新疆红井子盐碱土壤非培养放线菌多样性   总被引:1,自引:0,他引:1  
【目的】研究新疆红井子盐碱土壤中的放线菌物种多样性。【方法】应用基于16S rRNA基因序列系统发育分析的免培养方法进行放线菌物种多样性分析。利用放线菌特异性引物,以土壤样品总DNA为模板,扩增16S rRNA基因,构建16S rRNA基因克隆文库,并对文库中的插入序列进行RFLP分析。【结果】随机挑选的246个阳性克隆通过酶切筛选出61个不同图谱的重组克隆并测序。分析结果显示这61个克隆序列分属于42个OTUs,分布于放线菌纲(Actinobacteria)的放线菌亚纲(Actinobacteridae)、酸微菌亚纲(Acidimicrobidae)和红色杆菌亚纲(Rubrobacteridae);该环境中有71.4%的序列与已有效发表菌株的序列相似性小于97%,代表着放线菌新类群,其中部分序列形成了几个独立的进化分支,可能代表更高级的新分类单元。【结论】红井子土壤中的放线菌组成具有丰富的多样性,并有新放线菌分类单位的潜在资源,值得进一步进行开发研究。  相似文献   

7.
8.
Nematodes form an important component of many benthic marine ecosystems and DNA barcoding approaches could provide an insight into nematode community composition from different environments globally. We have amplified nematode 18S rRNA sequences using standard nematode18S rRNA primers from environmental DNA extracted from intertidal sediment collected from New Jersey coast, USA to test whether the published marine nematode 18S rRNA sequences from GenBank and EMBL databases can effectively assign unknown nematode sequences into genus or species level. Most of the sequenced clones showed some degree of identities with published marine nematode 18S rRNA sequences. However, relatively very few of the sequences could be assigned even to genus level based on sequence assignment rule. In addition, other eukaryotic 18S rRNA sequences were found to be co-amplified with commonly used nematode 18S rRNA primers. We found that the majority of the current nematode 18S rRNA primers will co-amplify other eukaryotes if environmental DNA is the target template. We therefore designed a new set of nematode 18S rRNA primers and evaluated them using environmental DNA in intertidal sediment from the New Jersey coast. In total, 40 clones were screened and subsequently sequenced and all the sequences showed varying degree of identities with published nematode 18S rRNA sequences from GenBank and EMBL databases, and no obvious eukaryotic co-amplicons were detected with new primers. Only 13 out of 40 clones amplified with the new primer set showed 100% identity to published Daptonema and Metachromadora 18S rRNA sequences. The current molecular databases for nematodes are dominated by sequences from NW Europe and need to be more extensively populated with new full length 18S rRNA nematode sequences collected from different biogeographic locations. The new primers developed in this study, in combination with an updated nematode 18S rRNA sequence database, would help us to better investigate and understand the diversity and community composition of free-living marine nematodes based on DNA barcoding approaches during biodiversity or biomonitoring surveys on a global-scale.  相似文献   

9.
Natural populations of benthic cyanobacteria in the lagoon of Tikehau Atoll in French Polynesia were studied using a polyphasic approach that combined field observations, LM, culturing, and molecular sequencing. The approach assessed their phenotypic (morphotypic and ecological) and genotypic diversity. Partial sequences (approximately 450 bp long) of the PCR-amplified 16S rRNA gene were obtained from both natural and cultured populations using cyanobacteria-specific primers. The sequences obtained clustered into six separate phylogenetic clusters in relation to the complete set of 16S rRNA sequences available in public databases. Phylogenetic clustering correlated in many cases with some morphological characteristics. For example, Spirulina subsalsa could be identified to the morphospecies level using both molecular and microscopic approaches. Morphotypes identified as Symploca hydnoides (Kütz. ex Gomont) TK22, Phormidium sp. TK1, and P. laysanense (Lemmerman) TK14 formed a distinct cluster. The morphogenus Hydrocoleum (Blennothrix) clustered interestingly close to the morphologically similar, but planktonic, marine cyanobacterium Trichodesmium , suggesting a relationship. Other sequences belonging to morphotaxa with very narrow trichomes were found to be polyphyletic. Enrichment cultures, with inoculum obtained from field populations, were followed over a period of 18 months. Dominance in all cultures shifted over time in favor of a set of cyanobacterial strains with narrow trichomes, which were phenotypically and phylogenetically different from natural populations dominating the original samples. Sequences from strains enriched by cultivation clustered into two well-defined phylogenetic groups, possibly identifying new taxa. These clusters may represent a niche of opportunistic species, evolved to exploit short-term nutrient spikes in the environment.  相似文献   

10.
Microeukaryotes in oxygen-depleted environments are among the most diverse, as well as the least studied, organisms. We conducted a cultivation-independent, small-subunit (SSU) rRNA-based survey of microeukaryotes in suboxic waters and anoxic sediments in the great Sippewisset salt marsh, Cape Cod, Mass. We generated two clone libraries and analyzed approximately 300 clones, which contained a large diversity of microeukaryotic SSU rRNA signatures. Only a few of these signatures were closely related (sequence similarity of >97%) to the sequences reported earlier. The bulk of our sequences represented deep novel branches within green algae, fungi, cercozoa, stramenopiles, alveolates, euglenozoa and unclassified flagellates. In addition, a significant number of detected rRNA sequences exhibited no affiliation to known organisms and sequences and thus represent novel lineages of the highest taxonomical order, most of them branching off the base of the global phylogenetic tree. This suggests that oxygen-depleted environments harbor diverse communities of novel organisms, which may provide an interesting window into the early evolution of eukaryotes.  相似文献   

11.
Several Thermus strains were isolated from 10 hot springs of the Rehai geothermal area in Tengchong, Yunnan province. The diversity of Thermus strains was examined by sequencing the 16S rRNA genes and comparing their sequences. Phylogenetic analysis showed that the 16S rDNA sequences from the Rehai geothermal isolates form four branches in the phylogenetic tree and had greater than 95.9% similarity in the phylogroup. Secondary structure comparison also indicated that the 16S rRNA from the Rehai geothermal isolates have unique secondary structure characteristics in helix 6, helix 9, and helix 10 (reference to Escherichia coli). This research is the first attempt to reveal the diversity of Thermus strains that are distributed in the Rehai geothermal area.  相似文献   

12.
云南江城和黑井盐矿沉积物未培养放线菌多样性比较   总被引:1,自引:0,他引:1  
类群特异性引物的应用使得研究者可以对感兴趣的微生物类群进行针对性研究.围绕云南江城和黑井两个地区的3个盐矿样点沉积物中放线菌的多样性和群落组成,我们通过放线菌特异性引物对总DNA进行16S rRNA基因扩增,经过克隆文库构建,利用酶切并选择其中不同带型的133个克隆的16S rRNA基因插入片段进行测序.系统发育分析和统计学结果表明,两地放线菌16S rRNA基因克隆广泛分布于整个放线菌门,同时发现部分序列可能属于放线菌的新类群.分析结果还预示,江城和黑井两地盐矿虽处云南不同地域含盐区,但两地未培养放线菌物种多样性和系统发育关系均较为相似.  相似文献   

13.
南海深海沉积物放线菌多样性分析   总被引:1,自引:0,他引:1  
【目的】免培养和纯培养相结合分析南海深海沉积物放线菌多样性。【方法】免培养方法通过提取沉积物宏基因组DNA,利用放线菌门特异性引物扩增放线菌16S r RNA基因序列,构建放线菌16S r RNA基因克隆文库,文库经RFLP(Restriction fragment length polymorphism)分析后挑选代表序列测序并进行多样性指数分析和系统发育分析。可培养方法利用8种培养基进行菌株分离,对排重后的菌株进行16S r RNA基因序列多样性分析。【结果】构建的两个深海位点的16S r RNA基因克隆文库在放线菌门的放线菌纲(Actinobacteria)、酸微菌纲(Acidimicrobiia)、腈基降解菌纲(Nitriliruptoria)和嗜热油菌纲(Thermoleophilia)4个纲中均有分布;两个位点中的种群结构有差异,N40-4位点的优势种群是放线菌纲的链霉菌目(Streptomycetales);N63-4位点的优势种群是腈基降解菌纲的腈基降解菌目(Nitriliruptorales)。8种培养基共分离出41株放线菌,根据形态特征排重后得到的19株菌分布于10个不同的属,12个不同的种,其中稀有放线菌属比例较高,菌株OAct400为潜在的微杆菌属(Microbacterium)新种。【结论】南海深海沉积物蕴含着丰富的放线菌物种资源及大量未知种群,具有进一步研究的价值。  相似文献   

14.
The phylogenetic diversity of the intestinal bacterial community in pigs was studied by comparative 16S ribosomal DNA (rDNA) sequence analysis. Samples were collected from a total of 24 pigs representing a variety of diets, ages, and herd health status. A library comprising 4,270 cloned 16S rDNA sequences obtained directly by PCR from 52 samples of either the ileum, the cecum, or the colon was constructed. In total, 375 phylotypes were identified using a 97% similarity criterion. Three hundred nine of the phylotypes (83%) had a <97% sequence similarity to any sequences in the database and may represent yet-uncharacterized bacterial genera or species. The phylotypes were affiliated with 13 major phylogenetic lineages. Three hundred four phylotypes (81%) belonged to the low-G+C gram-positive division, and 42 phylotypes (11.2%) were affiliated with the Bacteroides and Prevotella group. Four clusters of phylotypes branching off deeply within the low-G+C gram-positive bacteria and one in the Mycoplasma without any cultured representatives were found. The coverage of all the samples was 97.2%. The relative abundance of the clones approximated a lognormal distribution; however, the phylotypes detected and their abundance varied between two libraries from the same sample. The results document that the intestinal microbial community is very complex and that the majority of the bacterial species colonizing the gastrointestinal tract in pigs have not been characterized.  相似文献   

15.
Six matured male Yaks (Bos grunniens) with a mean live weight of 450 ± 23 kg (mean ± SD), were housed indoors in metabolism cages and fed pelleted lucerne (Medicago sativum). After an adjustment period of 24 days of feeding the diet, samples of rumen content were obtained for analysis of the bacteria in the liquor. The diversity of rumen bacteria was investigated by constructing a 16S rRNA gene clone library using the general bacterial primers F27 and R1492. A total of 130 clones, comprising nearly full length sequences (approx. 1.5 kb) were sequenced and submitted to BLAST and phylogenetic analysis. Using the criterion that similarity of 97% or greater with the sequences of cultivated bacteria, 16 clones were identified as Butyrivibrio fibrisolvens, Pseudobutyrivibrio ruminis, Ruminococcus flavefaciens, Succiniclasticum ruminis, Selenomonas ruminantium and Prevotella ruminicola, respectively. A further 10 clones shared similarity ranging from 90 to 97% with cultivated bacteria but the similarity in sequences for the remaining 104 clones were less than 90% of those of cultivated bacteria. Using a phylogenetic analysis it was found that the majority of the clones identified (63.8%) were located in the Low G + C Subdivision, with most of the remainder (35.4% of clones) located in the Cytophaga-Flexibacter-Bacteroides phylum and one clone (0.8%) was identified as a Proteobacteria. It was apparent that Yaks have a large and diverse range of bacteria in the rumen content which differ from those of cattle and other ruminants.  相似文献   

16.
Bacterial community structure and diversity in rhizospheres in two types of grassland, distinguished by both plant species and fertilization regimen, were assessed by performing a 16S ribosomal DNA (rDNA) sequence analysis of DNAs extracted from triplicate soil plots. PCR products were cloned, and 45 to 48 clones from each of the six libraries were partially sequenced. Phylogenetic analysis of the resultant 275 clone sequences indicated that there was considerable variation in abundance in replicate unfertilized, unimproved soil samples and fertilized, improved soil samples but that there were no significant differences in the abundance of any phylogenetic group. Several clone sequences were identical in the 16S rDNA region analyzed, and the clones comprised eight pairs of duplicate clones and two sets of triplicate clones. Many clones were found to be most closely related to environmental clones obtained in other studies, although three clones were found to be identical to culturable species in databases. The clones were clustered into operational taxonomic units at a level of sequence similarity of >97% in order to quantify diversity. In all, 34 clusters containing two or more sequences were identified, and the largest group contained nine clones. A number of diversity, dominance, and evenness indices were calculated, and they all indicated that diversity was high, reflecting the low coverage of rDNA libraries achieved. Differences in diversity between sample types were not observed. Collector's curves, however, indicated that there were differences in the underlying community structures; in particular, there was reduced diversity of organisms of the alpha subdivision of the class Proteobacteria (alpha-proteobacteria) in improved soils.  相似文献   

17.
During 1994 net sediment-water fluxes of oxygen, ammonium and inorganic phosphorus as well as sediment profiles of organic matter, nitrogen, phosphorus and iron were determined in three shallow eutrophic environments. Investigations were conducted monthly from March to December at five stations in the Sacca di Goro (Po River Delta, Italy). In the late summer, samples were collected from a single site in the Prévost lagoon (French Mediterranean coast) and three stations in the Bassin d'Arcachon (French Atlantic coast). In the Sacca di Goro, water-sediment exchanges of O2, NH 4 + and PO 4 3– were estimated by means of core incubation in the dark. Benthic fluxes for the French lagoons were in part determined experimentally using benthic chambers and in part from the literature.In general in the Sacca di Goro the highest oxygen uptake and nutrient release rates were found at the central sites, affected by macroalgal growth. At the sampling site adjacent to the freshwater inlet, sediment-water exchanges were principally influenced by tidal activity. In terms of organic matter and nutrient levels, sediments from the Sacca di Goro and from the Prévost lagoon, both colonised by the floating macroalga Ulva rigida C. Agardh, were similar. Sediments from the inner sheltered site in the Bassin d'Arcachon, invaded by the rooted macrophyte Ruppia cirrhosa (Pet.) Grande, showed the highest total N and P content (363 ± 157 µmol N cm–3 and 15 ± 2 µmol P cm–3 as average values in the top 10 cm of sediment), but were low in pore water ammonium and orthophosphate probably due to the high sequestering capacity of the system and/or efficient coupling between bacterial nutrient regeneration and assimilation by the plant roots. In addition the outer tidal stations in the Bassin d'Arcachon, invaded by rooted macrophytes, were low in pore water nutrients. A different trend was evident in the Prévost lagoon where the concentrations of exchangeable inorganic phosphorus and ammonium were appreciable (0.28 ± 0.07 µmol P cm–3 and 2.4 ± 1.4 µmol N cm–3 as average values in the top 10 cm of sediment). High amounts of dissolved organic nitrogen were found in the pore water at all the sites investigated showing the key role of the organic nitrogen in the recycling of nitrogen in these systems.The hypothesis that iron is a key factor in controlling phosphorus release is discussed since the Sacca di Goro, which is subject to dystrophic crises, is richer in iron than the Bassin d'Arcachon, which is a more buffered system.  相似文献   

18.
The diversity of sulfate-reducing bacteria (SRB) in brackish sediment was investigated using small-subunit rRNA and dissimilatory sulfite reductase (DSR) gene clone libraries and cultivation. The phylogenetic affiliation of the most commonly retrieved clones for both genes was strikingly similar and produced Desulfosarcina variabilis-like sequences from the inoculum but Desulfomicrobium baculatum-like sequences from a high dilution in natural media. Related organisms were subsequently cultivated from the site. PCR bias appear to be limited (or very similar) for the two primersets and target genes. However, the DSR primers showed a much higher phylogenetic specificity. DSR gene analysis is thus a promising and specific approach for investigating SRB diversity in complex habitats.  相似文献   

19.
To provide insight into the phylogenetic bacterial diversity of the freshwater sponge Spongilla lacustris, a 16S rRNA gene libraries were constructed from sponge tissues and from lake water. Restriction fragment length polymorphism (RFLP) analysis of >190 freshwater sponge-derived clones resulted in six major restriction patterns, from which 45 clones were chosen for sequencing. The resulting sequences were affiliated with the Alphaproteobacteria (n = 19), the Actinobacteria (n = 15), the Betaproteobacteria (n = 2), and the Chloroflexi (n = 2) lineages. About half of the sequences belonged to previously described actinobacterial (hgc-I) and betaproteobacterial (beta-II) sequence clusters of freshwater bacteria that were also present in the lake water 16S rRNA gene library. At least two novel, deeply rooting alphaproteobacterial lineages were recovered from S. lacustris that showed <89% sequence similarity to known phylogenetic groups. Electron microscopical observations revealed that digested bacterial remnants were contained within food vacuoles of sponge archaeocytes, whereas the extracellular matrix was virtually free of bacteria. This study is the first molecular diversity study of a freshwater sponge and adds to a growing database on the diversity and community composition of sponge-associated microbial consortia.  相似文献   

20.
Cyanobacterial 16S ribosomal RNA gene diversity was examined in a benthic mat on Fildes Peninsula of King George Island (62o09′54.4′′S, 58o57′20.9′′W), maritime Antarctica. Environmental DNA was isolated from the mat, a clone library of PCR-amplified 16S rRNA gene fragments was prepared, and amplified ribosomal DNA restriction analysis (ARDRA) was done to assign clones to seven groups. Low cyanobacterial diversity in the mat was suggested in that 83% of the clones were represented by one ARDRA group. DNA sequences from this group had high similarity with 16S rRNA genes of Tychonema bourrellyi and T. bornetii isolates, whose geographic origins were southern Norway and Northern Ireland. Cyanobacterial morphotypes corresponding to Tychonema have not been reported in Antarctica, however, this morphotype was previously found at Ward Hunt Lake (83oN), and in western Europe (52oN). DNA sequences of three of the ARDRA groups had highest similarity with 16S rDNA sequences of the Tychonema group accounting for 9.4% of the clones. Sequences of the remaining three groups (7.6%) had highest similarity with 16S rRNA genes of uncultured cyanobacteria clones from benthic mats of Lake Fryxell and fresh meltwater on the McMurdo Ice Shelf.  相似文献   

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