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1.
Int J Syst Evol Microbiol ; 65(12): 4776-4787, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26432704

ABSTRACT

Propionibacterium acnes subsp. acnes subsp. nov. and Propionibacterium acnes subsp. elongatum subsp. nov. are described. These emanate from the three known phylotypes of P. acnes, designated types I, II and III. Electron microscopy confirmed the filamentous cell shape of type III, showing a striking difference from types I/II, which were short rods. Biochemical tests indicated that, in types I/II, either the pyruvate, l-pyrrolidonyl arylamidase or d-ribose 2 test was positive, whereas all of these were negative among type III strains. Matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) spectra, which profile mainly their ribosomal proteins, were different between these two groups. Surface-enhanced laser-desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) spectra of all phylotypes revealed a specific protein biomarker that was overexpressed in type III strains compared with types I/II only when grown aerobically. Reference strains had high whole-genome similarity between types I (>91 %) and II (>75 %), but a considerably lower level of 72 % similarity with type III. recA and gyrB sequence dendrograms confirmed the distant relatedness of type III, indicating the presence of two distinct centres of variation within the species P. acnes. On the other hand, cellular fatty acid profiles and 16S rRNA gene sequence relatedness (>99.3 %) circumscribed the species. Thus, we propose two subspecies, Propionibacterium acnes subsp. acnes subsp. nov. for types I/II and Propionibacterium acnes subsp. elongatum subsp. nov. for type III. The type strain of Propionibacterium acnes subsp. acnes is NCTC 737T ( = ATCC 6919T = JCM 6425T = DSM 1897T = CCUG 1794T), while the type strain of Propionibacterium acnes subsp. elongatum is K124T ( = NCTC 13655T = JCM 18919T).


Subject(s)
Phylogeny , Propionibacterium acnes/classification , Bacterial Proteins/chemistry , Bacterial Typing Techniques , DNA, Bacterial/genetics , Fatty Acids/chemistry , Genes, Bacterial , Humans , Molecular Sequence Data , Nucleic Acid Hybridization , RNA, Ribosomal, 16S/genetics , Ribosomal Proteins/chemistry , Sequence Analysis, DNA , Skin/microbiology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
2.
J Clin Microbiol ; 53(1): 35-42, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25339391

ABSTRACT

The genus Yersinia is a large and diverse bacterial genus consisting of human-pathogenic species, a fish-pathogenic species, and a large number of environmental species. Recently, the phylogenetic and population structure of the entire genus was elucidated through the genome sequence data of 241 strains encompassing every known species in the genus. Here we report the mining of this enormous data set to create a multilocus sequence typing-based scheme that can identify Yersinia strains to the species level to a level of resolution equal to that for whole-genome sequencing. Our assay is designed to be able to accurately subtype the important human-pathogenic species Yersinia enterocolitica to whole-genome resolution levels. We also report the validation of the scheme on 386 strains from reference laboratory collections across Europe. We propose that the scheme is an important molecular typing system to allow accurate and reproducible identification of Yersinia isolates to the species level, a process often inconsistent in nonspecialist laboratories. Additionally, our assay is the most phylogenetically informative typing scheme available for Y. enterocolitica.


Subject(s)
Genome, Bacterial , Multilocus Sequence Typing , Yersinia Infections/microbiology , Yersinia/classification , Yersinia/genetics , Animals , Computational Biology/methods , Genes, Bacterial , Genetic Loci , Genetic Variation , Humans , Multilocus Sequence Typing/methods , Phylogeny , Reproducibility of Results , Yersinia/isolation & purification
3.
PLoS One ; 9(11): e112967, 2014.
Article in English | MEDLINE | ID: mdl-25415318

ABSTRACT

Enteroaggregative E. coli (EAEC) is an established diarrhoeagenic pathotype. The association with virulence gene content and ability to cause disease has been studied but little is known about the population structure of EAEC and how this pathotype evolved. Analysis by Multi Locus Sequence Typing of 564 EAEC isolates from cases and controls in Bangladesh, Nigeria and the UK spanning the past 29 years, revealed multiple successful lineages of EAEC. The population structure of EAEC indicates some clusters are statistically associated with disease or carriage, further highlighting the heterogeneous nature of this group of organisms. Different clusters have evolved independently as a result of both mutational and recombination events; the EAEC phenotype is distributed throughout the population of E. coli.


Subject(s)
Escherichia coli Infections/microbiology , Escherichia coli/genetics , Escherichia coli/pathogenicity , Multilocus Sequence Typing/methods , Bangladesh , Escherichia coli/classification , Escherichia coli Proteins/genetics , Evolution, Molecular , Humans , Mutation , Nigeria , Phylogeny , Polymerase Chain Reaction , Recombination, Genetic , Species Specificity , United Kingdom , Virulence/genetics
4.
J Med Microbiol ; 61(Pt 5): 622-630, 2012 May.
Article in English | MEDLINE | ID: mdl-22245790

ABSTRACT

Propionibacterium acnes, a commensal of human skin, is also an opportunistic pathogen of common acne and certain infectious diseases. However, it is still not obvious which strain is pathogenic for a certain infectious disease, and investigations to characterize pathogenic strains using molecular typing methods such as MLST using several housekeeping genes have been undertaken. However, to date, such analysis has focused mainly on strains isolated from Europeans, and it is unclear whether the clonal distribution in other parts of the world is similar. Here, we analysed 50 strains of P. acnes from healthy humans and patients with atopic dermatitis (AD) in Japan and utilized MLST of seven housekeeping genes to study their clonal patterns. The MLST successfully typed the strains into five types, IA, IB1, IB2, II and III. Strains that belonged to types IA, IB and II were common on the human skin of both populations (Europe and Japan), but this study demonstrated what we believe to be the first association of type III strains with human skin, existing on the skin of both the AD and non-AD population. These results indicate the global existence of type III strains on human skin.


Subject(s)
Genetic Variation , Multilocus Sequence Typing , Propionibacterium acnes/classification , Propionibacterium acnes/isolation & purification , Skin/microbiology , Europe , Gram-Positive Bacterial Infections/microbiology , Human Experimentation , Humans , Japan , Molecular Epidemiology , Propionibacterium acnes/genetics , Skin Diseases, Bacterial/microbiology
5.
Anaerobe ; 12(2): 71-4, 2006 Apr.
Article in English | MEDLINE | ID: mdl-16701617

ABSTRACT

Enterotoxigenic Bacteroides fragilis (ETBF) has been implicated in diarrhoeal illness in animals and humans. Recent data suggest that ETBF is associated with flares of inflammatory bowel disease. Toxigenicity is attributed to expression of a toxin referred to as fragilysin, which stimulates fluid accumulation in ligated intestinal segments and alter the morphology of human intestinal cells. Three different isoforms or variants of the enterotoxin gene, designated bft-1, bft-2, and bft-3, have been identified. In this study we investigated the distribution of bft alleles among ETBF strains in stool specimens from patients with colon cancer (n: 31), the control patients (n: 8) and extraintestinal sources (n: 15). We used restriction fragment length polymorphism analysis of the PCR-amplified enterotoxin gene and sequencing the PCR-product to detect the isoforms of bft gene. Among the stool strains, bft-1 was found to be more common than bft-2; as it was detected 27 of 31 strains from colon cancer patients and 7 of 8 control strains. The bft-1 isoform was also found in almost all isolates from extraintestinal sites. No bft-3 subtype was detected among all tested strains.


Subject(s)
Bacterial Toxins/genetics , Bacteroides Infections/microbiology , Bacteroides fragilis/genetics , Feces/microbiology , Metalloendopeptidases/genetics , Adult , Aged , Aged, 80 and over , Alleles , Bacterial Toxins/classification , Bacteroides Infections/complications , Bacteroides fragilis/isolation & purification , Case-Control Studies , Colonic Neoplasms/complications , Colonic Neoplasms/microbiology , DNA Primers/chemistry , DNA, Bacterial/chemistry , Humans , Metalloendopeptidases/classification , Middle Aged , Polymerase Chain Reaction/methods , Polymorphism, Restriction Fragment Length
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