Phylogenetic study of Staphylococcus and Macrococcus species based on partial hsp60 gene sequences. (1/49)

A 600 bp partial hsp60 gene sequence has been described previously as a novel genetic marker for species identification and phylogenetic studies within the genus Staphylococcus. In the present study, the 600 bp partial hsp60 gene sequences of 40 validly described Staphylococcus species and subspecies and four Macrococcus species were PCR-amplified and sequenced. Phylogenetic analysis revealed excellent concordance between the unrooted dendrograms based on partial hsp60 and 16S rRNA gene sequences. The genus Macrococcus is clearly separated from the genus Staphylococcus, but is closely related to the 'sciuri group', the only staphylococci that are cytochrome c oxidase-positive. The remaining Staphylococcus species clustered into five broad-based subdivisions, which corresponded to the 'aureus group', the 'epidermidis group', the 'haemolyticus group', the 'saprophyticus group' and the 'intermedius group'. These results agreed remarkably well with the current taxonomy of this diverse family, which is based on classical phenotypic and biochemical testing. Furthermore, pairwise sequence comparisons indicated that the hsp60 gene is more divergent and more discriminatory than the 16S rRNA gene for species differentiation among strains of the genera Staphylococcus and Macrococcus. It is concluded that the hsp60 gene may be an efficient alternative target for taxonomic and phylogenetic studies on members of these genera.  (+info)

Jeotgalicoccus halotolerans gen. nov., sp. nov. and Jeotgalicoccus psychrophilus sp. nov., isolated from the traditional Korean fermented seafood jeotgal. (2/49)

Two Gram-positive, non-motile, non-spore-forming, halotolerant and moderately halophilic cocci (strains YKJ-101T and YKJ-115T) were isolated from the traditional Korean fermented seafood jeotgal, and were investigated using a polyphasic taxonomic approach. Phylogenetic analysis of 16S rDNA sequences showed that strains YKJ-101T and YKJ-115T are most closely related to the cluster comprising two Salinicoccus species. The peptidoglycan type of the strains is A3alpha, based on L-Lys-Gly(3-4)-L-Ala(Gly), and the predominant menaquinone is MK-7. Strains YKJ-101T and YKJ-115T have cellular fatty acid profiles containing major amounts of saturated, unsaturated and branched fatty acids; the major fatty acids are anteiso-C15 : 0 and iso-C15 : 0. The cellular polar lipids are phosphatidylglycerol, diphosphatidylglycerol and unidentified phospholipids. Strains YKJ-101T and YKJ-115T have identical DNA G + C contents of 42 mol%. The 16S rDNA similarity between strains YKJ-101T and YKJ-115T is 98% and the mean level of DNA-DNA relatedness between the two strains is 13.4%. On the basis of phenotypic and phylogenetic data and genomic distinctiveness, it is proposed that strains YKJ-101T and YKJ-115T should be placed in a new genus, Jeotgalicoccus gen. nov., as two distinct new species, for which the names Jeotgalicoccus halotolerans sp. nov. and Jeotgalicoccus psychrophilus sp. nov. are proposed. The type strains are YKJ-101T (=KCCM 41448T =JCM 11198T) and YKJ-115T (=KCCM 41449T =JCM 11199T), respectively.  (+info)

Gemella bacteraemia characterised by 16S ribosomal RNA gene sequencing. (3/49)

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.  (+info)

Macrococcus brunensis sp. nov., Macrococcus hajekii sp. nov. and Macrococcus lamae sp. nov., from the skin of llamas. (4/49)

Eight strains of Gram-positive, catalase- and oxidase-positive cocci were isolated from the skin of llamas (Lama glama L.) and characterized using a polyphasic approach. These strains were assigned to the genus Macrococcus on the basis of their phenotypic properties (resistance to bacitracin and sensitivity to furazolidone) and DNA base content (40-42 mol% G+C). Phylogenetic analysis based on 16S rDNA confirmed that the strains are members of the genus MACROCOCCUS: They differed from all hitherto described macrococcal species in their production of phosphatase and reduction of nitrate (most strains) and the inability to produce acid from glycerol or to grow in 7.5 % NaCl. Ribotyping (EcoRI), macrorestriction analysis (XbaI) and fatty acid methyl ester analysis divided the strains from llamas into three stable clusters. Moreover, ribotyping differentiated the strains analysed not only from previously described macrococcal species but also from oxidase-positive staphylococci. DNA-DNA hybridization confirmed that the three clusters represent separate genomic groups (similarity values<54 %). All the results showed that the strains represent three novel species, for which the names Macrococcus hajekii sp. nov. (type strain CCM 4809(T)=LMG 21711(T)), Macrococcus brunensis sp. nov. (type strain CCM 4811(T)=LMG 21712(T)) and Macrococcus lamae sp. nov. (type strain CCM 4815(T)=LMG 21713(T)) are proposed.  (+info)

rpoB gene sequence-based identification of aerobic Gram-positive cocci of the genera Streptococcus, Enterococcus, Gemella, Abiotrophia, and Granulicatella. (5/49)

We developed a new molecular tool based on rpoB gene (encoding the beta subunit of RNA polymerase) sequencing to identify streptococci. We first sequenced the complete rpoB gene for Streptococcus anginosus, S. equinus, and Abiotrophia defectiva. Sequences were aligned with these of S. pyogenes, S. agalactiae, and S. pneumoniae available in GenBank. Using an in-house analysis program (SVARAP), we identified a 740-bp variable region surrounded by conserved, 20-bp zones and, by using these conserved zones as PCR primer targets, we amplified and sequenced this variable region in an additional 30 Streptococcus, Enterococcus, Gemella, Granulicatella, and Abiotrophia species. This region exhibited 71.2 to 99.3% interspecies homology. We therefore applied our identification system by PCR amplification and sequencing to a collection of 102 streptococci and 60 bacterial isolates belonging to other genera. Amplicons were obtained in streptococci and Bacillus cereus, and sequencing allowed us to make a correct identification of streptococci. Molecular signatures were determined for the discrimination of closely related species within the S. pneumoniae-S. oralis-S. mitis group and the S. agalactiae-S. difficile group. These signatures allowed us to design a S. pneumoniae-specific PCR and sequencing primer pair.  (+info)

Brain abscess due to Gemella haemolysans. (6/49)

We present a case of brain abscess due to Gemella haemolysans and Bacteroides species in a 60-year-old-immunocompetent man who underwent dental procedures. The patient completely recovered following intravenous therapy with ampicillin and metronidazole for 6 weeks.  (+info)

Jeotgalicoccus pinnipedialis sp. nov., from a southern elephant seal (Mirounga leonina). (7/49)

A previously unknown Gram-positive, catalase-positive, facultatively anaerobic, non-spore-forming, coccus-shaped bacterium (A/G14/99/10(T)), originating from the mouth of a female southern elephant seal, was subjected to a taxonomic analysis. Comparative 16S rRNA gene-sequencing showed that the organism formed a hitherto unknown subline within the catalase-positive, low-G+C, Gram-positive cocci, exhibiting a specific association with species of the genus Jeotgalicoccus. Sequence divergence values of approximately 7 %, together with phenotypic differences, showed the unknown bacterium to be distinct from the two described species of this genus, Jeotgalicoccus halotolerans and Jeotgalicoccus psychrophilus. Based on phenotypic and phylogenetic considerations, it is proposed that strain A/G14/99/10(T)=CCUG 42722(T)=CIP 107946(T) from the mouth of a seal be classified as the type strain of a novel species of the genus Jeotgalicoccus, Jeotgalicoccus pinnipedialis sp. nov.  (+info)

Phenotypic and molecular characterization of erythromycin resistance in four isolates of Streptococcus-like gram-positive cocci causing bacteremia. (8/49)

Among nine patients with bacteremia caused by Granulicatella or Gemella in a 6-year period (July 1995 to June 2001), three had bacteremia caused by erythromycin-resistant Granulicatella adiacens and one had bacteremia caused by erythromycin-resistant Gemella haemolysans. All four isolates possessed mef genes, whereas none possessed ermT, ermTR, or ermB genes.  (+info)