Home About us Contact
|
Home About us Contact | ||
|
|
||
rDNA Clone Library (rdna + clone_library)
Selected AbstractsCrash of a population of the marine heterotrophic flagellate Cafeteria roenbergensis by viral infectionENVIRONMENTAL MICROBIOLOGY, Issue 11 2007Ramon Massana Summary Viruses are known as important mortality agents of marine microorganisms. Most studies focus on bacterial and algal viruses, and few reports exist on viruses infecting marine heterotrophic protists. Here we show results from several incubations initiated with a microbial assemblage from the central Indian Ocean and amended with different amounts of organic matter. Heterotrophic flagellates developed up to 30 000 cells ml,1 in the most enriched incubation. A 18S rDNA clone library and fluorescent in situ hybridization counts with newly designed probes indicated that the peak was formed by Cafeteria roenbergensis and Caecitellus paraparvulus (90% and 10% of the cells respectively). Both taxa were below detection in the original sample, indicating a strong positive selective bias during the enrichment. During the peak, C. roenbergensis cells were observed with virus-like particles in the cytoplasm, and 4 days later this taxa could not be detected. Transmission electron microscopy confirmed the viral nature of these particles, which were large (280 nm), had double-stranded DNA, and were produced with a burst size of ,70. This virus was specific of C. roenbergensis as neither C. paraparvulus that was never seen infected, nor other flagellate taxa that developed in later stages of the incubation, appeared attacked. This is one of the few reports on a heterotrophic flagellate virus and the implications of this finding in the Indian Ocean are discussed. [source] High bacterial diversity of a waste gas-degrading community in an industrial biofilter as shown by a 16S rDNA clone libraryENVIRONMENTAL MICROBIOLOGY, Issue 11 2002Udo Friedrich Summary The bacterial diversity of an industrial biofilter used for waste gas abatement in an animal-rendering plant was investigated. A 16S rDNA clone library was generated and 444 clones were screened using computer-aided amplified ribosomal DNA restriction analysis (ARDRA). Of the screened clones, 60.8% showed unique ARDRA patterns and the remaining 174 clones were clustered into 65 groups. Almost full-length 16S rDNA sequences of 106 clones were determined and 90.5% of the clones were affiliated with the two phyla Proteobacteria and Bacteroidetes. Alpha -, Beta -, and Gammaproteobacteria accounted for 22.1, 17.6 and 18.6% respectively. Minor portions were affiliated with the Actinobacteria (2.0%), Firmicutes and Verrucomicrobia (both 1.0%), and the Deltaproteobacteria and Thermomicrobia (each 0.5%). Only six out of the 106 16S rDNA sequences exhibited similarities of more than 97% to classified bacterial species indicating that a substantial fraction of the clone sequences were derived from unknown taxa. It was also evaluated whether a database containing 281 computer-simulated bacterial rDNA fragment patterns generated from published reference sequences can be used for identification purposes. The data analysis demonstrated that this was possible only for a small number of clones, which were closely related to described bacterial strains. Rarefaction analysis of ARDRA clusters demonstrated that the 444 clones screened are insufficient to describe the entire diversity of the clone library. [source] Genetic diversity of the toxic cyanobacterium Microcystis in Lake MikataENVIRONMENTAL TOXICOLOGY, Issue 3 2005Mitsuhiro Yoshida Abstract The aim of the present study was to clarify the bloom dynamics and community composition of hepatotoxin microcystin-producing and non-microcystin-producing Microcystis genotypes in the environment. In Lake Mikata (Fukui, Japan) from April 2003 to January 2004, seasonal variation in the number of cells with microcystin (mcy) genotypes and the genetic diversity of the total population were investigated using quantitative competitive PCR and a 16S rDNA clone library, respectively. Using competitive PCR, cells with mcyA genotypes were quantified in August and October, and the ratio of the number of these mcyA genotypes to colony-forming Microcystis cells was 0.37 and 2.37, respectively. The 16S rDNA clones obtained could be divided into 12 ribotypes: a,l. Sixty-one Microcystis strains isolated from Lake Mikata during the sampling period were subjected to toxicity tests using HPLC and ELISA, PCR-based detection of the mcyA gene, and sequence analysis of the 16S rDNA. All isolates could be differentiated into 11 ribotypes (a, b, d, f, h, i, and m,q). Ribotypes b, f, i, m, n, and p had at least one strain that was a microcystin producer. In natural communities ribotypes b and f accounted for 85% of the 16S rDNA clones in August, and ribotypes b and i accounted for 24% of the clones in October. Thus, in some bloom stages the presence of microcystin genotypes identified using the 16S rDNA clone library correlated with that of mcy genotypes determined using competitive PCR. © 2005 Wiley Periodicals, Inc. Environ Toxicol 20: 229,234, 2005. [source] High prokaryote diversity and analysis of community structure in mobile mud deposits off French Guiana: identification of two new bacterial candidate divisionsFEMS MICROBIOLOGY ECOLOGY, Issue 3 2001Vanessa M. Madrid Abstract Bacterial and archaeal community compositions in highly mobile nearshore muds typical of the Guiana coastline of South America were examined by sequence analysis of a 16S rDNA clone library. DNA was extracted from a subsurface sediment layer (10,30 cm) collected at a subtidal (,1 m deep) mud wave site between Kourou and Sinnamary, French Guiana. Analysis of 96 non-chimeric sequences showed the majority to be bacteria (98%), that diversity was high with 64 unique sequences, and that proteobacteria were dominant (46%). Two crenarchaeota sequences were found (2%). Bacterial sequences belonged to the Cytophaga-Flexibacter-Bacteroides (18%), Actinobacteria (11.5%), Planctomycetes (6.3%), Cyanobacteria (3.2%), low-GC Gram-positive (1%), ,, , and , subdivisions of Proteobacteria (27%, 16%, and 9%, respectively). Additional bacterial sequences belonged to the candidate division TM6 (1%) and to two newly proposed candidate divisions: KS-A (2%) and KS-B (3%). A sizeable fraction (22%) of sequences from the Kourou,Sinnamary library are normally found in water column populations, reflecting frequent entrainment of suspended debris into physically reworked underlying sediments. Dominant sequences (56%) were related to Gelidibacter algens (Cytophaga-Flexibacter-Bacteroides group), Actinobacteria, Sulfitobacter and Ruegeria spp. (,-proteobacteria), all of which are chemoorganotrophs, consistent with abundant labile organic carbon. The presence of sequences from potential sulfate reducers and sulfide oxidizers suggests the likelihood of sulfur cycling in these sediments, despite the dominance of suboxic (iron-reducing), non-sulfidic diagenetic properties. Rarefaction analysis indicated that bacterial diversity in the French Guiana library is not only unusually high in comparison with other marine sedimentary environments, but among the most diverse of all environments reported to date. [source] Diversity and abundance of Bacteria and Archaea in the Bor Khlueng Hot Spring in ThailandJOURNAL OF BASIC MICROBIOLOGY, Issue 6 2004Pattanop Kanokratana The prokaryotic diversity in the Bor Khlueng hot spring in Ratchaburi province, Thailand was investigated by a culture-independent molecular approach. This hydrothermal pool is located in the central part of Thailand and contains sulfide-rich mineral water that is believed to relieve muscle ache and pain. The water flow year-round with temperature ranging between 50,57 °C. Community DNA was extracted directly from sediment samples by coring to depth of ,20,30 cm. Small-subunit rRNA genes (16S rDNA) were amplified by PCR using primers specific for the domains Archaea and Bacteria. The PCR products were cloned and sequenced. For the bacterial rDNA clone library, 200 clones were randomly selected for further analyses. After restriction fragment length polymorphism (RFLP) analysis of rDNA clones and exclusion of chimeric sequences 36 phylotypes were obtained. The Bor Khlueng phylotypes spanned a wide range within the domain Bacteria, occupying eleven major lineages (phyla). Almost a quarter (23%) of the clones were classifed as Acidobacteria. The other clones were grouped into the Bacteriodetes (19%), Nitrospirae (13%), Proteobacteria (12%), Deinococcus-Thermus lineage (11%), planctomycetes (6%), and Verrucomicrobia (5%). The four remaining phyla, 5% each, were assigned to Actinobacteria, Chloroflexi, Cyanobacteria, and the candidate division "OP10". For the archaeal 16S rRNA gene sequence library, 25 distinct phylotypes were obtained, 17 clones were found to be associated with Crenarahaeota and 8 clones were associated with Euryarachaeota. The findings of the molecular survey of this so far not investigated site showed that Bor Khlueng hot spring is a potential rich source of unique bacterial and archaeal species. The great majority (,80%) of the prokaryotic sequences detected in Bor Khlueng were unknown. (© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Marine sponge Craniella austrialiensis -associated bacterial diversity revelation based on 16S rDNA library and biologically active Actinomycetes screening, phylogenetic analysisLETTERS IN APPLIED MICROBIOLOGY, Issue 4 2006Z.-Y. Li Abstract Aims:, The aim of this study was to investigate the bacterial diversity associated with the sponge Craniella australiensis using a molecular strategy and isolating Actinomycetes with antimicrobial potentials. Methods and Results:, The bacterial diversity associated with South China Sea sponge C. austrialiensis was assessed using a 16S rDNA clone library alongside restriction fragment length polymorphism and phylogenetic analysis. It was found that the C. austrialiensis -associated bacterial community consisted of alpha, beta and gamma- Proteobacteria, Firmicutes, Bacteroidetes as well as Actinobacterium. Actinomycetes were isolated successfully using seawater medium with sponge extracts. According to the BLAST and phylogenetic analysis based on about 600-bp 16S rDNA sequences, 11 of the representative 23 isolates closely matched the Streptomyces sp. while the remaining 12 matched the Actinomycetales. Twenty Actinomycetes have antimicrobial potentials, of which 15 are found to possess broad-spectrum antimicrobial potentials. Conclusions:, The sponge C. austrialiensis -associated bacterial community is very abundant including Proteobacteria, Firmicutes, Bacteroidetes and Actinobacterium while Actinomycetes is not predominant. Artificial seawater medium with sponge extracts is suitable for Actinomycetes isolation. Most of the isolated C. austrialiensis -associated Actinomycetes have a broad spectrum of antimicrobial activity. Significance and Impact of the Study:, This study revealed the diversity of the bacterial community and the isolated Actinomycetes with antimicrobial potentials associated with sponge C. australiensis. [source] Phylogenetic analysis of the fecal flora of the wild pygmy lorisAMERICAN JOURNAL OF PRIMATOLOGY, Issue 8 2010Xu Bo Abstract The bacterial diversity in fecal samples from the wild pygmy loris was examined with a 16S rDNA clone library and restriction fragment length polymorphism analysis. The clones were classified as Firmicutes (43.1%), Proteobacteria (34.5%), Actinobacteria (5.2%), and Bacteroidetes (17.2%). The 58 different kinds of 16S rDNA sequences were classified into 16 genera and 20 uncultured bacteria. According to phylogenetic analysis, the major genera within the Proteobacteria was Pseudomonas, comprising 13.79% of the analyzed clone sequences. Many of the isolated rDNA sequences did not correspond to known microorganisms, but had high homology to uncultured clones found in human feces. Am. J. Primatol. 72:699,706, 2010. © 2010 Wiley-Liss, Inc. [source] Effect of the applied organic load rate on biodegradable polymer production by mixed microbial cultures in a sequencing batch reactorBIOTECHNOLOGY & BIOENGINEERING, Issue 1 2006Davide Dionisi Abstract This article studies the operation of a new process for the production of biopolymers (polyhydroxyalkanoates, PHAs) at different applied organic load rates (OLRs). The process is based on the aerobic enrichment of activated sludge to obtain mixed cultures able to store PHAs at high rates and yields. A mixture of acetic, lactic, and propionic acids at different concentrations (in the range 8.5,31.25 gCOD/L) was fed every 2 h in a sequencing batch reactor (SBR). The resulting applied OLR was in the range 8.5,31.25 gCOD/L/day. Even though, as expected, the increase in the OLR caused an increase in biomass concentration (up to about 8.7 g COD/L), it also caused a relevant decrease of maximal polymer production rate. This decrease in polymer production rate was related to the different extent of "feast and famine" conditions, as function of the applied OLR and of the start-up conditions. As a consequence the best performance of the process was obtained at an intermediate OLR (20 gCOD/L/day) where both biomass productivity and PHA storage were high enough. However, at this high OLR the process was unstable and sudden decrease of performance was also observed. The sludge characterized by the highest PHA storage response was investigated by 16S rDNA clone library. The clone library contained sequences mostly from PHA producers (e.g., Alcaligenes and Comamonas genera); however many genera and among them, one of the dominant (Thauera), were never described before in relation to PHA storage response. © 2005 Wiley Periodicals, Inc. [source] | ||||||||||