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PubMed2004

Prevotella shahii sp. nov. and Prevotella salivae sp. nov., isolated from the human oral cavity.

Two bacterial strains, EHS11(T) and EPSA11(T), which were isolated from the human oral cavity, were characterized in terms of phenotypic and biochemical characteristics, cellular fatty acid profiles and phylogenetic position based on 16S rRNA gene sequence analysis. 16S rRNA gene sequence analysis showed that each of the isolates belonged to a novel species of the genus Prevotella. Strain EHS11(T) was related to Prevotella loescheii (about 95 % similarity), whereas strain EPSA11(T) was related to Prevotella oris (about 94 % similarity). Both strains were obligately anaerobic, non-pigmented, non-spore-forming, non-motile, Gram-negative rods. The cellular fatty acid composition of strain EPSA11(T) was very similar to that of P. oris JCM 8540(T). On the other hand, the cellular fatty acid composition of strain EHS11(T) was significantly different from those of other Prevotella species. The predominant fatty acids in strain EHS11(T) are C(18 : 1)omega9c, C(16 : 0) and C(16 : 0) 3-OH, whereas other Prevotella species, except for P. loescheii JCM 8530(T), possess anteiso-C(15 : 0), iso-C(17 : 0) 3-OH and C(18 : 1)omega9c. The predominant fatty acids in P. loescheii JCM 8530(T) are anteiso-C(15 : 0), C(16 : 0) and C(18 : 1)omega9c. DNA-DNA hybridization experiments revealed a genomic distinction of strains EHS11(T) and EPSA11(T) from P. loescheii JCM 8530(T) and P. oris JCM 8540(T). On the basis of these data, two novel Prevotella species are proposed: Prevotella shahii sp. nov. and Prevotella salivae sp. nov. The type strains of P. shahii and P. salivae are EHS11(T) (=JCM 12083(T)=DSM 15611(T)) and EPSA11(T) (=JCM 12084(T)=DSM 15606(T)), respectively.

DNA, Bacterial↗
PubMed2003

The phylogeny of Paralabrax (Perciformes: Serranidae) and allied taxa inferred from partial 16S and 12S mitochondrial ribosomal DNA sequences.

Partial sequences of 16S and 12S mitochondrial ribosomal DNA were used to examine the phylogenetic relationships of the primarily eastern Pacific genus Paralabrax (Perciformes: Serranidae) and allied taxa. Paralabrax is considered a basal serranine, which is itself considered the basal subfamily in the Serranidae. Multiple serranines reported closely related to Paralabrax from the genera Serranus, Hypoplectrus, Cratinus, and Centropristis were used as outgroups. Species from the remaining two subfamilies, Epinephilinae and Anthiinae, of the Serranidae were also used in the analyses. The tree of the Serranidae was rooted with the families Polyprionidae and Priacanthidae. Paralabrax, the Serranidae, and the Serraninae were monophyletic in this study. Serranus was found to be paraphyletic. Centropristis, formerly considered the sister taxon to Paralabrax, was not closely related in these analyses. Cratinus agassizii, a monotypic genus from the eastern Pacific, was found to be the sister taxon to Paralabrax. There is greater resolution for intergeneric and subfamily relations than interspecific relationships. A single most parsimonious tree for the interspecific relationships of Paralabrax and allied taxa is proposed. This proposed molecular phylogeny is consistent with known biogeographic processes in the eastern Pacific.

Animals↗
PubMed2005

The hierarchical system of the 'Alphaproteobacteria': description of Hyphomonadaceae fam. nov., Xanthobacteraceae fam. nov. and Erythrobacteraceae fam. nov.

Phylogenetic analysis of the class 'Alphaproteobacteria', including physiologically diverse species, was conducted by using small-subunit rRNA gene sequences. The 16S rRNA gene sequences of 261 species in the class 'Alphaproteobacteria' were obtained from GenBank/EMBL/DDBJ for constructing a phylogenetic tree by using maximum-likelihood analysis. In the resulting tree, members of the class 'Alphaproteobacteria' were subdivided into five major clusters, which were compared with the taxonomic outline of Bergey's Manual of Systematic Biology and the arb tree. Based on this phylogenetic tree, three novel families are proposed: Hyphomonadaceae fam. nov. to accommodate the bacterial genera Hyphomonas, Hirschia, Maricaulis and Oceanicaulis, Xanthobacteraceae fam. nov. to include the genera Xanthobacter, Azorhizobium, Ancylobacter, Labrys and Starkeya, and Erythrobacteraceae fam. nov. to accommodate the genera Erythrobacter, Porphyrobacter and Erythromicrobium. The phylogenetic tree of 16S rRNA gene sequences established in this study may provide a sound basis for future taxonomic reconstruction of the class 'Alphaproteobacteria'.

Alphaproteobacteria↗
PubMed1993

A phylogeny of New World Deltocephalus-like leafhopper genera based on mitochondrial 16S ribosomal DNA sequences.

The Deltocephalus-like leafhoppers are a group of grass-feeding insects that share common forms in the male genitalia, chiefly a fused aedeagal-connective structure. The group contains 27 New World genera. The phylogenetic relationships among these genera and other genera in the tribe are unclear because fusion of the aedeagus and connective, although clearly apomorphic, has occurred in both related and unrelated groups. An independent set of characters is required to test the hypothesis that the Deltocephalus-like genera are monophyletic and to derive relationships among these genera. We examined 562 sites in the 3'-end of the mitochondrial 16S ribosomal DNA from 21 species representing 19 New World genera. Eight species in closely related groups or tribes were also analyzed. The region was sequenced directly from PCR-amplified DNA of field-collected and museum-preserved dried specimens. The transversion rate was twice as high as the transition rate; however, 74% of these were adenine-thymine transversions. Analysis of secondary structure indicated that substitution rates were equal in stem and loop forming regions of the processed rRNA. Using either parsimony or distance analysis, derived phylogenies suggested that, with the exception of the genus Cabrulus, the Deltocephalus-like leafhoppers are genera within a monophyletic group.

Animals↗
PubMed1992

Identification of xenobiotic-degrading isolates from the beta subclass of the Proteobacteria by a polyphasic approach including 16S rRNA partial sequencing.

Nineteen gram-negative, aerobic, biodegradative isolates were identified by using a polyphasic taxonomic approach. The presence of the specific polyamine 2-hydroxyputrescine and the presence of a ubiquinone with eight isoprenoid units in the side chain (ubiquinone Q-8) allowed allocation of these organisms to the beta subclass of the Proteobacteria. On the basis of the results of additional characterization experiments (i.e., API 20NE tests, determinations of soluble protein patterns, and DNA-DNA hybridization experiments), we classified six isolates as either Comamonas testosteroni, Comamonas acidovorans, or Alcaligenes xylosoxidans subsp. denitrificans. By using the same criteria we allocated two additional isolates to the genus Alcaligenes. A comparison of a 16S rRNA fragment (positions 1220 to 1377; Escherichia coli nomenclature) indicated that the remaining isolates should be allocated as follows: one is a member of C. testosteroni and one is a member of Acidovorax facilis, as confirmed by the results of additional DNA-DNA hybridizations; two others probably belong to the family Alcaligenaceae; six are related to "Alcaligenes eutrophus"; and one, strain NRRL 12228, occupies an isolated position.

Bacterial Proteins↗
PubMed2004

Lancefield group C Streptococcus dysgalactiae infection responsible for fish mortalities in Japan.

A Lancefield serological group C Streptococcus sp. was isolated from cultured amberjack, Seriola dumerili Risso, and yellowtail, Seriola quinqueradiata Temminck and Schlegel, immunized with Lactococcus garvieae commercial vaccines in Japan. The isolated bacteria were Gram-positive cocci, auto-aggregating in saline, morphologically long chains in growth medium, catalase negative and alpha-haemolytic on blood agar. An almost complete gene sequence of the 16S rDNA of two isolates was determined and compared with that of bacterial strains in the database. The isolates were identified as Streptococcus dysgalactiae based on the results of the 16S rDNA sequence, the bacteriological properties and the Lancefield serological grouping. Oligonucleotide primers specifically designed for the 16S-23S rDNA intergenic spacer region of S. dysgalactiae amplified a gene from all the fish isolates, as well as the type strains alpha-haemolytic S. dysgalactiae subsp. dysgalactiae ATCC430738 and beta-haemolytic S. dysgalactiae subsp. equisimilis ATCC35666, but not those of S. equi ATCC33398, Lactococcus garvieae ATCC43921 and L. garvieae KG9408. The severe necrotic lesions of the caudal peduncle seen in experimentally infected fish were similar to those seen in naturally infected fish.

Animals↗
PubMed2005

RAPD & FINS (mitochondrial 16s rRNA gene sequence) in analysis of food contamination: detection of food poisoning by Lizard.

In poisoning, detection of the nature of causative agent is important for management of trauma and forensic investigation. Most of the methods in clinical toxicology are developed for detection of toxins and poisons. A random amplified polymorphic DNA (RAPD)-based method has been described for detection of species of animal from its morphologically unrecognizable fragments, recovered from food substances, consumption of which caused even death. Pre-mixed RAPD reaction beads and six RAPD primers were used in polymerase chain reaction analysis. Among six RAPD primers used, any one of them was sufficient in resolving this practical forensic situation. To enhance the probability values for matching in the present study of fixing identity of an animal, six set of market available RAPD primers were used. This is the first report of a forensic application of RAPD DNA typing in identification of charred skeleton remnants of Lizard species in food material. Furthermore unique amplicons were generated for different reptilian species, which can be used as species specific markers for species identification in forensic situation, however, no variations among individuals of same species were observed.

Animals↗
PubMed2003

Gemella bacteraemia characterised by 16S ribosomal RNA gene sequencing.

AIMS: To define epidemiology, clinical disease, and outcome of gemella bacteraemia by 16S rRNA gene sequencing. To examine the usefulness of the Vitek, API, and ATB systems in identifying two gemella species. METHODS: All alpha haemolytic streptococci other than Streptococcus pneumoniae isolated from blood cultures during a six year period were identified by conventional biochemical methods, the Vitek system, and the API system. 16S rRNA gene sequencing was performed on all isolates identified by both kits as gemella with >or= 95% confidence or by either kit as any bacterial species with < 95% confidence. The ATB expression system was used to identify the two isolates that were defined as gemella species by 16S rRNA gene sequencing. RESULTS: Of the 302 alpha haemolytic streptococci other than S pneumoniae isolated, one was identified as Gemella morbillorum, and another as Gemella haemolysans by 16S rRNA gene sequencing. The patient with monomicrobial G morbillorum bacteraemia was a 66 year old man with community acquired infective endocarditis with septic thromboemboli. The patient with G haemolysans bacteraemia was a 41 year old woman with hospital acquired polymicrobial bacteraemia during the neutropenic period of an autologous bone marrow transplant for non-Hodgkin's lymphoma, the first case of its kind in the English literature. The API and ATB expression systems only identified the second strain as G haemolysans at 94% and 99% confidence, respectively, whereas the Vitek system identified none of the two strains correctly at > 70% confidence. CONCLUSIONS: Gemella bacteraemia is uncommon. 16S rRNA gene sequencing is the method of choice for identification of gemella and gemella-like isolates.

Adult↗
PubMed2004

Clinical and laboratory features of Mycobacterium porcinum.

Recent molecular studies have shown Mycobacterium porcinum, recovered from cases of lymphadenitis in swine, to have complete 16S rDNA sequence identity and >70% DNA-DNA homology with human isolates within the M. fortuitum third biovariant complex. We identified 67 clinical and two environmental isolates of the M. fortuitum third biovariant sorbitol-negative group, of which 48 (70%) had the same PCR restriction enzyme analysis (PRA) profile as the hsp65 gene of M. porcinum (ATCC 33776(T)) and were studied in more detail. Most U.S. patient isolates were from Texas (44%), Florida (19%), or other southern coastal states (15%). Clinical infections included wound infections (62%), central catheter infections and/or bacteremia (16%), and possible pneumonitis (18%). Sequencing of the 16S rRNA gene (1,463 bp) showed 100% identity with M. porcinum ATCC 33776(T). Sequencing of 441 bp of the hsp65 gene showed four sequevars that differed by 2 to 3 bp from the porcine strains. Clinical isolates were positive for arylsulfatase activity at 3 days, nitrate, iron uptake, D-mannitol, i-myo-inositol, and catalase at 68 degrees C. They were negative for L-rhamnose and D-glucitol (sorbitol). Clinical isolates were susceptible to ciprofloxacin, sulfamethoxazole, and linezolid and susceptible or intermediate to cefoxitin, clarithromycin, imipenem, and amikacin. M. porcinum ATCC 33776(T) gave similar results except for being nitrate negative. These studies showed almost complete phenotypic and molecular identity between clinical isolates of the M. fortuitum third biovariant D-sorbitol-negative group and porcine strains of M. porcinum and confirmed that they belong to the same species. Identification of M. porcinum presently requires hsp65 gene PRA or 16S rRNA or hsp65 gene sequencing.

Animals↗
PubMed1982

Sequence of the chloroplast 16S rRNA gene and its surrounding regions of Chlamydomonas reinhardii.

The sequence of a 2 kb DNA fragment containing the chloroplast 16S ribosomal RNA gene from Chlamydomonas reinhardii and its flanking regions has been determined. The algal 16S rRNA sequence (1475 nucleotides) and secondary structure are highly related to those found in bacteria and in the chloroplasts of higher plants. In contrast, the flanking regions are very different. In C. reinhardii the 16S rRNA gene is surrounded by AT rich segments of about 180 bases, which are followed by a long stretch of complementary bases separated from each other by 1833 nucleotides. It is likely that these structures play an important role in the folding and processing of the precursor of 16S rRNA. The primary and secondary structures of the binding sites of two ribosomal proteins in the 16SrRNAs of E. coli and C. reinhardii are considerably related.

Base Composition↗
PubMed2001

Phylogenetic relationships of Xylella fastidiosa strains from different hosts, based on 16S rDNA and 16S-23S intergenic spacer sequences.

The phylogenetic relationships of Xylella fastidiosa strains isolated from different hosts, including citrus trees, coffee, grapevine, plum and pear, were inferred by sequence analysis of the 16S rDNA and 16S-23S intergenic spacer region. A high level of similarity (97.1-100%) was found in the 16S rDNA of the Xylella fastidiosa strains. The 16S-23S region showed a higher level of variation, with similarity values ranging from 79.8 to 100%. Two tRNAs (tRNA(Ala) and tRNA(Ile)) were encountered within the spacer sequence. The phylogenetic trees, constructed using the neighbour-joining method, showed that the citrus, coffee, peach and plum strains were closely related and separate from grapevine strains. The pear strain remained isolated from all the other Xylella strains in both analyses and produced values of less than 20% in DNA-DNA hybridization experiments with a citrus strain. These results show that this strain does not belong to the Xylella fastidiosa genomic species.

DNA, Bacterial↗
PubMed2005

Phylogenetic relationships of the tribe Paini (Amphibia, Anura, Ranidae) based on partial sequences of mitochondrial 12s and 16s rRNA genes.

Partial sequences of mitochondrial 12S and 16S rRNA genes from 19 Asian frog species of the tribe Paini (Ranidae, Dicroglossinae) allowed a first molecular study of the phylogenetic relationships of this tribe. This analysis confirmed that this tribe is a monophyletic group, but suggested relationships did not agree with previous generic classification of this clade based on morphology. Two major clades were recognized within the Paini. For one of them, the generic name Quasipaa is available. Phylogenetic relationships within the other group are not yet fully clarified and need further study.

Animals↗
PubMed2006

Utility of ITS1-5.8S-ITS2 and 16S mitochondrial DNA sequences for species identification and phylogenetic inference within the Rhinonyssus coniventris species complex (Acari: Rhinonyssidae).

The complete internal transcribed spacer 1 (ITS1), 5.8S rDNA, and ITS2 region of the ribosomal DNA and a 390-bp region of the 16S rDNA gene from five taxa belonging to Rhinonyssus (Rhinonyssus vanellus, Rhinonyssus tringae, Rhinonyssus neglectus, Rhinonyssus echinipes from Kentish plover, and Rhinonyssus echinipes from grey plover) were sequenced to examine the level of sequence variation and the taxonomic levels to show utility in phylogeny estimation. Our data show that these molecular markers can help to discriminate between species and populations included in the Rhinonyssus coniventris complex (R. tringae, R. neglectus, R. echinipes), which are morphologically very close and difficult to separate by classic methods. A comparative study with sequences from other rhinonyssid mites previously published was also carried out. The resulting phylogenetic tree inferred from ITS1-5.8S-ITS2 region sequences obtained in this paper, together with those from other 11 taxa of rhinonyssid, shows slight differences from the current taxonomy of the Rhinonyssidae. This study appeals for the revision of the taxonomic status of the R. coniventris complex, as well as for the species included within it.

Animals↗
PubMed2006

Changes in the phyllosphere community of the resurrection fern, Polypodium polypodioides, associated with rainfall and wetting.

A combination of analyses were used to characterize the changes that occur in a bacterial community present in the phyllosphere of the epiphytic resurrection fern, Polypodium polypodioides, as the fern rehydrates from a desiccation-resistant, physiologically inactive state. Enrichment assays showed an increase in the viable count of bacteria using labile organic substrates following rainfall. Isolates obtained from enrichments were predominantly Gram-positive bacteria affiliated with various groups of the Actinobacteria and Firmicutes. In contrast, sequencing of 16S rRNA genes clones obtained from whole community DNA revealed that much of the community was dominated by other taxa, particularly the Alphaproteobacteria. Similar isolates were obtained from both dry and hydrated P. polypodioides fronds, whereas 16S rRNA gene sequencing of community DNA revealed different ribotypes on the dry and wet fern, and an overall reduction in richness following wetting. Wetting also produced changes in phyllosphere extracellular enzyme activity, with an initial burst of activity following rainfall and a subsequent burst approximately 48 h later. These findings suggest that the resurrection fern harbors a complex phyllosphere community, and that rehydration of the fern following rainfall may act as an enrichment culture stimulating certain bacterial populations and changing the overall community structure and activity.

Acid Phosphatase↗