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1.
J Clin Microbiol ; 56(6)2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29643197

RESUMO

Listeria monocytogenes causes foodborne outbreaks with high mortality. For improvement of outbreak cluster detection, the German consiliary laboratory for listeriosis implemented whole-genome sequencing (WGS) in 2015. A total of 424 human L. monocytogenes isolates collected in 2007 to 2017 were subjected to WGS and core-genome multilocus sequence typing (cgMLST). cgMLST grouped the isolates into 38 complexes, reflecting 4 known and 34 unknown disease clusters. Most of these complexes were confirmed by single nucleotide polymorphism (SNP) calling, but some were further differentiated. Interestingly, several cgMLST cluster types were further subtyped by pulsed-field gel electrophoresis, partly due to phage insertions in the accessory genome. Our results highlight the usefulness of cgMLST for routine cluster detection but also show that cgMLST complexes require validation by methods providing higher typing resolution. Twelve cgMLST clusters included recent cases, suggesting activity of the source. Therefore, the cgMLST nomenclature data presented here may support future public health actions.


Assuntos
Variação Genética , Genoma Bacteriano , Listeria monocytogenes/classificação , Listeriose/epidemiologia , Técnicas de Tipagem Bacteriana , Bacteriófagos/genética , Análise por Conglomerados , Eletroforese em Gel de Campo Pulsado , Genótipo , Alemanha/epidemiologia , Humanos , Listeria monocytogenes/isolamento & purificação , Tipagem de Sequências Multilocus , Filogenia , Polimorfismo de Nucleotídeo Único , Sorogrupo , Sequenciamento Completo do Genoma
2.
Food Microbiol ; 71: 46-54, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29366468

RESUMO

In Germany salmonellosis still represents the 2nd most common bacterial foodborne disease. The majority of infections are caused by Salmonella (S.) Typhimurium and S. Enteritidis followed by a variety of other broad host-range serovars. Salmonella Derby is one of the five top-ranked serovars isolated from humans and it represents one of the most prevalent serovars in pigs, thus bearing the potential risk for transmission to humans upon consumption of pig meat and products thereof. From November 2013 to January 2014 S. Derby caused a large outbreak that affected 145 primarily elderly people. Epidemiological investigations identified raw pork sausage as the probable source of infection, which was confirmed by microbiological evidence. During the outbreak isolates from patients, food specimen and asymptomatic carriers were investigated by conventional typing methods. However, the quantity and quality of available microbiological and epidemiological data made this outbreak highly suitable for retrospective investigation by Whole Genome Sequencing (WGS) and subsequent evaluation of different bioinformatics approaches for cluster definition. Overall the WGS-based methods confirmed the results of the conventional typing but were of significant higher discriminatory power. That was particularly beneficial for strains with incomplete epidemiological data. For our data set both, single nucleotide polymorphism (SNP)- and core genome multilocus sequence typing (cgMLST)-based methods proved to be appropriate tools for cluster definition.


Assuntos
Doenças Transmitidas por Alimentos/microbiologia , Infecções por Salmonella/microbiologia , Salmonella enterica/isolamento & purificação , Animais , DNA Bacteriano/genética , Surtos de Doenças , Doenças Transmitidas por Alimentos/epidemiologia , Genoma Bacteriano , Alemanha/epidemiologia , Humanos , Produtos da Carne/microbiologia , Tipagem de Sequências Multilocus , Filogenia , Polimorfismo de Nucleotídeo Único , Carne Vermelha/microbiologia , Estudos Retrospectivos , Infecções por Salmonella/epidemiologia , Salmonella enterica/classificação , Salmonella enterica/genética , Sorogrupo , Suínos , Sequenciamento Completo do Genoma
3.
4.
BMC Genomics ; 16 Suppl 5: S7, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26041051

RESUMO

BACKGROUND: Corynebacterium urealyticum is an opportunistic pathogen that normally lives on skin and mucous membranes in humans. This high Gram-positive bacteria can cause acute or encrusted cystitis, encrusted pyelitis, and pyelonephritis in immunocompromised patients. The bacteria is multi-drug resistant, and knowledge about the genes that contribute to its virulence is very limited. Two complete genome sequences were used in this comparative genomic study: C. urealyticum DSM 7109 and C. urealyticum DSM 7111. RESULTS: We used comparative genomics strategies to compare the two strains, DSM 7109 and DSM 7111, and to analyze their metabolic pathways, genome plasticity, and to predict putative antigenic targets. The genomes of these two strains together encode 2,115 non-redundant coding sequences, 1,823 of which are common to both genomes. We identified 188 strain-specific genes in DSM 7109 and 104 strain-specific genes in DSM 7111. The high number of strain-specific genes may be a result of horizontal gene transfer triggered by the large number of transposons in the genomes of these two strains. Screening for virulence factors revealed the presence of the spaDEF operon that encodes pili forming proteins. Therefore, spaDEF may play a pivotal role in facilitating the adhesion of the pathogen to the host tissue. Application of the reverse vaccinology method revealed 19 putative antigenic proteins that may be used in future studies as candidate drug or vaccine targets. CONCLUSIONS: The genome features and the presence of virulence factors in genomic islands in the two strains of C. urealyticum provide insights in the lifestyle of this opportunistic pathogen and may be useful in developing future therapeutic strategies.


Assuntos
Hibridização Genômica Comparativa/métodos , Biologia Computacional/métodos , Corynebacterium/genética , Genoma Bacteriano/genética , Fatores de Virulência/genética , Corynebacterium/classificação , Corynebacterium/patogenicidade , Infecções por Corynebacterium/tratamento farmacológico , Infecções por Corynebacterium/microbiologia , Farmacorresistência Bacteriana Múltipla/genética , Genômica/métodos , Humanos
5.
J Bacteriol ; 194(16): 4437, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22843578

RESUMO

Bovine mastitis represents the most economically important disease in dairy cows and can be caused by Corynebacterium bovis, a commensal in the bovine udder. The draft genome sequence provides insights into the adaptation of this bacterium to the bovine habitat and its lipolytic capabilities to utilize components of cow's milk.


Assuntos
Corynebacterium/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Genoma Bacteriano , Análise de Sequência de DNA , Adaptação Biológica , Animais , Bovinos , Corynebacterium/isolamento & purificação , Metabolismo dos Lipídeos , Mastite Bovina/microbiologia , Redes e Vias Metabólicas/genética , Leite/metabolismo , Dados de Sequência Molecular
6.
J Bacteriol ; 194(12): 3199-215, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22505676

RESUMO

Corynebacterium diphtheriae is one of the most prominent human pathogens and the causative agent of the communicable disease diphtheria. The genomes of 12 strains isolated from patients with classical diphtheria, endocarditis, and pneumonia were completely sequenced and annotated. Including the genome of C. diphtheriae NCTC 13129, we herewith present a comprehensive comparative analysis of 13 strains and the first characterization of the pangenome of the species C. diphtheriae. Comparative genomics showed extensive synteny and revealed a core genome consisting of 1,632 conserved genes. The pangenome currently comprises 4,786 protein-coding regions and increases at an average of 65 unique genes per newly sequenced strain. Analysis of prophages carrying the diphtheria toxin gene tox revealed that the toxoid vaccine producer C. diphtheriae Park-Williams no. 8 has been lysogenized by two copies of the ω(tox)(+) phage, whereas C. diphtheriae 31A harbors a hitherto-unknown tox(+) corynephage. DNA binding sites of the tox-controlling regulator DtxR were detected by genome-wide motif searches. Comparative content analysis showed that the DtxR regulons exhibit marked differences due to gene gain, gene loss, partial gene deletion, and DtxR binding site depletion. Most predicted pathogenicity islands of C. diphtheriae revealed characteristics of horizontal gene transfer. The majority of these islands encode subunits of adhesive pili, which can play important roles in adhesion of C. diphtheriae to different host tissues. All sequenced isolates contain at least two pilus gene clusters. It appears that variation in the distributed genome is a common strategy of C. diphtheriae to establish differences in host-pathogen interactions.


Assuntos
Corynebacterium diphtheriae/genética , Corynebacterium diphtheriae/isolamento & purificação , Difteria/microbiologia , Endocardite Bacteriana/microbiologia , Variação Genética , Genoma Bacteriano , Pneumonia Bacteriana/microbiologia , DNA Bacteriano/química , DNA Bacteriano/genética , Deleção de Genes , Transferência Genética Horizontal , Genes Bacterianos , Ilhas Genômicas , Glicolipídeos/genética , Humanos , Dados de Sequência Molecular , Mutagênese Insercional , Prófagos/genética , Regulon , Análise de Sequência de DNA
7.
BMC Genomics ; 13: 141, 2012 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-22524407

RESUMO

BACKGROUND: Corynebacterium resistens was initially recovered from human infections and recognized as a new coryneform species that is highly resistant to antimicrobial agents. Bacteremia associated with this organism in immunocompromised patients was rapidly fatal as standard minocycline therapies failed. C. resistens DSM 45100 was isolated from a blood culture of samples taken from a patient with acute myelocytic leukemia. The complete genome sequence of C. resistens DSM 45100 was determined by pyrosequencing to identify genes contributing to multi-drug resistance, virulence, and the lipophilic lifestyle of this newly described human pathogen. RESULTS: The genome of C. resistens DSM 45100 consists of a circular chromosome of 2,601,311 bp in size and the 28,312-bp plasmid pJA144188. Metabolic analysis showed that the genome of C. resistens DSM 45100 lacks genes for typical sugar uptake systems, anaplerotic functions, and a fatty acid synthase, explaining the strict lipophilic lifestyle of this species. The genome encodes a broad spectrum of enzymes ensuring the availability of exogenous fatty acids for growth, including predicted virulence factors that probably contribute to fatty acid metabolism by damaging host tissue. C. resistens DSM 45100 is able to use external L-histidine as a combined carbon and nitrogen source, presumably as a result of adaptation to the hitherto unknown habitat on the human skin. Plasmid pJA144188 harbors several genes contributing to antibiotic resistance of C. resistens DSM 45100, including a tetracycline resistance region of the Tet W type known from Lactobacillus reuteri and Streptococcus suis. The tet(W) gene of pJA144188 was cloned in Corynebacterium glutamicum and was shown to confer high levels of resistance to tetracycline, doxycycline, and minocycline in vitro. CONCLUSIONS: The detected gene repertoire of C. resistens DSM 45100 provides insights into the lipophilic lifestyle and virulence functions of this newly recognized pathogen. Plasmid pJA144188 revealed a modular architecture of gene regions that contribute to the multi-drug resistance of C. resistens DSM 45100. The tet(W) gene encoding a ribosomal protection protein is reported here for the first time in corynebacteria. Cloning of the tet(W) gene mediated resistance to second generation tetracyclines in C. glutamicum, indicating that it might be responsible for the failure of minocycline therapies in patients with C. resistens bacteremia.


Assuntos
Corynebacterium/efeitos dos fármacos , Corynebacterium/genética , Resistência a Múltiplos Medicamentos/genética , Genoma Bacteriano , Aminoácidos/metabolismo , Proteínas de Bactérias/genética , Cromossomos Bacterianos , Corynebacterium/metabolismo , Ordem dos Genes , Histidina/metabolismo , Humanos , Leucemia Mieloide Aguda/microbiologia , Redes e Vias Metabólicas , Testes de Sensibilidade Microbiana , Anotação de Sequência Molecular , Dados de Sequência Molecular , Proteínas de Ligação às Penicilinas/genética , Plasmídeos/genética , Análise de Sequência de DNA , Fatores de Virulência/genética
8.
Microb Pathog ; 52(3): 165-76, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22239957

RESUMO

Although Corynebacterium diphtheriae has been classically described as an exclusively extracellular pathogen, there is growing evidence that it may be internalized by epithelial cells. The aim of the present report was to investigate the nature and involvement of the surface-exposed non-fimbrial 67-72 kDa proteins (67-72p), previously characterized as adhesin/hemagglutinin, in C. diphtheriae internalization by HEp-2 cells. Transmission electron microscopy and bacterial internalization inhibition assays indicated the role of 67-72p as invasin for strains of varied sources. Cytoskeletal changes with accumulation of polymerized actin in HEp-2 cells beneath adherent 67-72p-adsorbed microspheres were observed by the Fluorescent actin staining test. Trypan blue staining method and Methylthiazole tetrazolium reduction assay showed a significant decrease in viability of HEp-2 cells treated with 67-72p. Morphological changes in HEp-2 cells observed after treatment with 67-72p included vacuolization, nuclear fragmentation and the formation of apoptotic bodies. Flow cytometry revealed an apoptotic volume decrease in HEp-2 cells treated with 67-72p. Moreover, a double-staining assay using Propidium Iodide/Annexin V gave information about the numbers of vital vs. early apoptotic cells and late apoptotic or secondary necrotic cells. The comparative analysis of MALDI-TOF MS experiments with the probes provided for 67-72p CDC-E8392 with an in silico proteome deduced from the complete genome sequence of C. diphtheriae identified with significant scores 67-72p as the protein DIP0733. In conclusion, DIP0733 (67-72p) may be directly implicated in bacterial invasion and apoptosis of epithelial cells in the early stages of diphtheria and C. diphtheriae invasive infection.


Assuntos
Apoptose , Corynebacterium diphtheriae/patogenicidade , Endocitose , Hemaglutininas/metabolismo , Hepatócitos/microbiologia , Hepatócitos/fisiologia , Fatores de Virulência/metabolismo , Actinas/metabolismo , Linhagem Celular , Sobrevivência Celular , Corynebacterium diphtheriae/genética , Hemaglutininas/genética , Humanos , Multimerização Proteica , Fatores de Virulência/genética
9.
FEMS Yeast Res ; 12(3): 382-6, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22292503

RESUMO

The ascomycetous yeast Wickerhamomyces anomalus (formerly Pichia anomala and Hansenula anomala) exhibits antimicrobial activities and flavoring features that are responsible for its frequent association with food, beverage and feed products. However, limited information on the genetic background of this yeast and its multiple capabilities are currently available. Here, we present the draft genome sequence of the neotype strain W. anomalus DSM 6766. On the basis of pyrosequencing, a de novo assembly of this strain resulted in a draft genome sequence with a total size of 25.47 Mbp. An automatic annotation using RAPYD generated 11 512 protein-coding sequences. This annotation provided the basis to analyse metabolic capabilities, phylogenetic relationships, as well as biotechnologically important features and yielded novel candidate genes of W. anomalus DSM 6766 coding for proteins participating in antimicrobial activities.


Assuntos
Anti-Infecciosos/metabolismo , Biotecnologia/métodos , Genoma Fúngico/genética , Fatores Matadores de Levedura/metabolismo , Pichia/genética , Análise de Sequência de DNA/métodos , Compostos Orgânicos Voláteis/metabolismo , Mapeamento Cromossômico , Microbiologia de Alimentos , Proteínas Fúngicas/metabolismo , Anotação de Sequência Molecular , Dados de Sequência Molecular , Filogenia , Pichia/classificação , Pichia/metabolismo
10.
J Bacteriol ; 193(1): 323-4, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21037006

RESUMO

This work reports the completion and annotation of the genome sequence of Corynebacterium pseudotuberculosis I19, isolated from an Israeli dairy cow with severe clinical mastitis. To present the whole-genome sequence, a de novo assembly approach using 33 million short (25-bp) mate-paired SOLiD reads only was applied. Furthermore, the automatic, functional, and manual annotations were attained with the use of several algorithms in a multistep process.


Assuntos
Corynebacterium pseudotuberculosis/genética , Genoma Bacteriano , Mastite Bovina/microbiologia , Animais , Bovinos , Corynebacterium pseudotuberculosis/classificação , Corynebacterium pseudotuberculosis/isolamento & purificação , Feminino , Israel/epidemiologia , Mastite Bovina/epidemiologia , Dados de Sequência Molecular
11.
J Bacteriol ; 193(22): 6420-1, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22038974

RESUMO

In this work, we report the complete genome sequence of a Corynebacterium pseudotuberculosis PAT10 isolate, collected from a lung abscess in an Argentine sheep in Patagonia, whose pathogen also required an investigation of its pathogenesis. Thus, the analysis of the genome sequence offers a means to better understanding of the molecular and genetic basis of virulence of this bacterium.


Assuntos
Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/genética , Genoma Bacteriano , Abscesso Pulmonar/microbiologia , Doenças dos Ovinos/microbiologia , Animais , Argentina , Sequência de Bases , Corynebacterium pseudotuberculosis/isolamento & purificação , Corynebacterium pseudotuberculosis/patogenicidade , Dados de Sequência Molecular , Ovinos , Virulência
12.
J Bacteriol ; 193(24): 7025-6, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22123771

RESUMO

In this work, we report the whole-genome sequence of Corynebacterium pseudotuberculosis bv. equi strain CIP 52.97 (Collection Institut Pasteur), isolated in 1952 from a case of ulcerative lymphangitis in a Kenyan horse, which has evidently caused significant losses to agribusiness. Therefore, obtaining this genome will allow the detection of important targets for postgenomic studies, with the aim of minimizing problems caused by this microorganism.


Assuntos
Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/genética , Genoma Bacteriano , Doenças dos Cavalos/microbiologia , Animais , Sequência de Bases , Infecções por Corynebacterium/microbiologia , Corynebacterium pseudotuberculosis/isolamento & purificação , Cavalos , Quênia , Dados de Sequência Molecular
13.
BMC Genomics ; 12: 545, 2011 Nov 03.
Artigo em Inglês | MEDLINE | ID: mdl-22053731

RESUMO

BACKGROUND: Corynebacterium variabile is part of the complex microflora on the surface of smear-ripened cheeses and contributes to the development of flavor and textural properties during cheese ripening. Still little is known about the metabolic processes and microbial interactions during the production of smear-ripened cheeses. Therefore, the gene repertoire contributing to the lifestyle of the cheese isolate C. variabile DSM 44702 was deduced from the complete genome sequence to get a better understanding of this industrial process. RESULTS: The chromosome of C. variabile DSM 44702 is composed of 3, 433, 007 bp and contains 3, 071 protein-coding regions. A comparative analysis of this gene repertoire with that of other corynebacteria detected 1, 534 predicted genes to be specific for the cheese isolate. These genes might contribute to distinct metabolic capabilities of C. variabile, as several of them are associated with metabolic functions in cheese habitats by playing roles in the utilization of alternative carbon and sulphur sources, in amino acid metabolism, and fatty acid degradation. Relevant C. variabile genes confer the capability to catabolize gluconate, lactate, propionate, taurine, and gamma-aminobutyric acid and to utilize external caseins. In addition, C. variabile is equipped with several siderophore biosynthesis gene clusters for iron acquisition and an exceptional repertoire of AraC-regulated iron uptake systems. Moreover, C. variabile can produce acetoin, butanediol, and methanethiol, which are important flavor compounds in smear-ripened cheeses. CONCLUSIONS: The genome sequence of C. variabile provides detailed insights into the distinct metabolic features of this bacterium, implying a strong adaption to the iron-depleted cheese surface habitat. By combining in silico data obtained from the genome annotation with previous experimental knowledge, occasional observations on genes that are involved in the complex metabolic capacity of C. variabile were integrated into a global view on the lifestyle of this species.


Assuntos
Queijo/microbiologia , Corynebacterium/genética , Microbiologia de Alimentos , Genoma Bacteriano , Biologia Computacional , Corynebacterium/isolamento & purificação , Corynebacterium/metabolismo , DNA Bacteriano/genética , Ferro/metabolismo , Metaboloma , Anotação de Sequência Molecular , Análise de Sequência de DNA , Paladar
14.
BMC Genomics ; 12: 383, 2011 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-21801446

RESUMO

BACKGROUND: Corynebacterium ulcerans has been detected as a commensal in domestic and wild animals that may serve as reservoirs for zoonotic infections. During the last decade, the frequency and severity of human infections associated with C. ulcerans appear to be increasing in various countries. As the knowledge of genes contributing to the virulence of this bacterium was very limited, the complete genome sequences of two C. ulcerans strains detected in the metropolitan area of Rio de Janeiro were determined and characterized by comparative genomics: C. ulcerans 809 was initially isolated from an elderly woman with fatal pulmonary infection and C. ulcerans BR-AD22 was recovered from a nasal sample of an asymptomatic dog. RESULTS: The circular chromosome of C. ulcerans 809 has a total size of 2,502,095 bp and encodes 2,182 predicted proteins, whereas the genome of C. ulcerans BR-AD22 is 104,279 bp larger and comprises 2,338 protein-coding regions. The minor difference in size of the two genomes is mainly caused by additional prophage-like elements in the C. ulcerans BR-AD22 chromosome. Both genomes show a highly similar order of orthologous coding regions; and both strains share a common set of 2,076 genes, demonstrating their very close relationship. A screening for prominent virulence factors revealed the presence of phospholipase D (Pld), neuraminidase H (NanH), endoglycosidase E (EndoE), and subunits of adhesive pili of the SpaDEF type that are encoded in both C. ulcerans genomes. The rbp gene coding for a putative ribosome-binding protein with striking structural similarity to Shiga-like toxins was additionally detected in the genome of the human isolate C. ulcerans 809. CONCLUSIONS: The molecular data deduced from the complete genome sequences provides considerable knowledge of virulence factors in C. ulcerans that is increasingly recognized as an emerging pathogen. This bacterium is apparently equipped with a broad and varying set of virulence factors, including a novel type of a ribosome-binding protein. Whether the respective protein contributes to the severity of human infections (and a fatal outcome) remains to be elucidated by genetic experiments with defined bacterial mutants and host model systems.


Assuntos
Corynebacterium/genética , Genômica , Fatores de Virulência/genética , Idoso , Sequência de Aminoácidos , Animais , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Cães , Feminino , Ordem dos Genes , Genoma Bacteriano/genética , Humanos , Modelos Moleculares , Dados de Sequência Molecular , Prófagos/genética , Conformação Proteica , Fatores de Virulência/química
15.
Microb Genom ; 7(10)2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34693903

RESUMO

Non-typhoidal Salmonella enterica is an important gastrointestinal pathogen causing a considerable burden of disease. Resistance to third generation cephalosporins poses a serious threat for treatment of severe infections. In this study occurrence, phylogenetic relationship, and mechanisms of third generation cephalosporin resistance were investigated for clinical non-typhoidal S. enterica isolates in Germany. From 2017 to 2019, we detected 168 unique clinical S. enterica isolates with phenotypic resistance to third generation cephalosporins in a nation-wide surveillance. Compared to previous years, we observed a significant (P=0.0002) and consistent increase in resistant isolates from 0.41 % in 2005 to 1.71 % in 2019. In total, 34 different serovars were identified, most often S. Infantis (n=41; 24.4 %), S. Typhimurium (n=27; 16.1 %), S. Kentucky (n=21; 12.5 %), and S. Derby (n=17; 10.1 %). Whole genome analyses revealed extended-spectrum ß-lactamase (ESBL) genes as main cause for third generation cephalosporin resistance, and most prevalent were blaCTX-M-1 (n=55), blaCTX-M-14 (n=25), and blaCTX-M-65 (n=23). There was no strict correlation between serovar, phylogenetic lineage, and ESBL type but some serovar/ESBL gene combinations were detected frequently, such as blaCTX-M-1 and blaCTX-M-65 in S. Infantis or blaCTX-M-14b in S. Kentucky. The ESBL genes were mainly located on plasmids, including IncI, IncA/C variants, emerging pESI variants, and a novel blaCTX-M-1harbouring plasmid. We conclude that third generation cephalosporin resistance is on the rise among clinical S. enterica isolates in Germany, and occurrence in various S. enterica serovars is most probably due to multiple acquisition events of plasmids.


Assuntos
Resistência às Cefalosporinas/genética , Farmacorresistência Bacteriana Múltipla/genética , Plasmídeos/genética , Salmonella enterica/genética , beta-Lactamases/genética , Antibacterianos/farmacologia , Cefalosporinas , Alemanha , Humanos , Testes de Sensibilidade Microbiana , Filogenia , Salmonella enterica/classificação , Salmonella enterica/efeitos dos fármacos , Salmonella enterica/isolamento & purificação , Sorogrupo
16.
BMC Genomics ; 11: 91, 2010 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-20137072

RESUMO

BACKGROUND: Corynebacterium aurimucosum is a slightly yellowish, non-lipophilic, facultative anaerobic member of the genus Corynebacterium and predominantly isolated from human clinical specimens. Unusual black-pigmented variants of C. aurimucosum (originally named as C. nigricans) continue to be recovered from the female urogenital tract and they are associated with complications during pregnancy. C. aurimucosum ATCC 700975 (C. nigricans CN-1) was originally isolated from a vaginal swab of a 34-year-old woman who experienced a spontaneous abortion during month six of pregnancy. For a better understanding of the physiology and lifestyle of this potential urogenital pathogen, the complete genome sequence of C. aurimucosum ATCC 700975 was determined. RESULTS: Sequencing and assembly of the C. aurimucosum ATCC 700975 genome yielded a circular chromosome of 2,790,189 bp in size and the 29,037-bp plasmid pET44827. Specific gene sets associated with the central metabolism of C. aurimucosum apparently provide enhanced metabolic flexibility and adaptability in aerobic, anaerobic and low-pH environments, including gene clusters for the uptake and degradation of aromatic amines, L-histidine and L-tartrate as well as a gene region for the formation of selenocysteine and its incorporation into formate dehydrogenase. Plasmid pET44827 codes for a non-ribosomal peptide synthetase that plays the pivotal role in the synthesis of the characteristic black pigment of C. aurimucosum ATCC 700975. CONCLUSIONS: The data obtained by the genome project suggest that C. aurimucosum could be both a resident of the human gut and possibly a pathogen in the female genital tract causing complications during pregnancy. Since hitherto all black-pigmented C. aurimucosum strains have been recovered from female genital source, biosynthesis of the pigment is apparently required for colonization by protecting the bacterial cells against the high hydrogen peroxide concentration in the vaginal environment. The location of the corresponding genes on plasmid pET44827 explains why black-pigmented (formerly C. nigricans) and non-pigmented C. aurimucosum strains were isolated from clinical specimens.


Assuntos
Corynebacterium/genética , Genoma Bacteriano , Vagina/microbiologia , Aborto Espontâneo , Adulto , Biologia Computacional , Corynebacterium/crescimento & desenvolvimento , Infecções por Corynebacterium/microbiologia , DNA Bacteriano/genética , Feminino , Genes Bacterianos , Humanos , Família Multigênica , Gravidez , Análise de Sequência de DNA
17.
BMC Genomics ; 11: 728, 2010 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-21192786

RESUMO

BACKGROUND: Corynebacterium pseudotuberculosis is generally regarded as an important animal pathogen that rarely infects humans. Clinical strains are occasionally recovered from human cases of lymphadenitis, such as C. pseudotuberculosis FRC41 that was isolated from the inguinal lymph node of a 12-year-old girl with necrotizing lymphadenitis. To detect potential virulence factors and corresponding gene-regulatory networks in this human isolate, the genome sequence of C. pseudotuberculosis FCR41 was determined by pyrosequencing and functionally annotated. RESULTS: Sequencing and assembly of the C. pseudotuberculosis FRC41 genome yielded a circular chromosome with a size of 2,337,913 bp and a mean G+C content of 52.2%. Specific gene sets associated with iron and zinc homeostasis were detected among the 2,110 predicted protein-coding regions and integrated into a gene-regulatory network that is linked with both the central metabolism and the oxidative stress response of FRC41. Two gene clusters encode proteins involved in the sortase-mediated polymerization of adhesive pili that can probably mediate the adherence to host tissue to facilitate additional ligand-receptor interactions and the delivery of virulence factors. The prominent virulence factors phospholipase D (Pld) and corynebacterial protease CP40 are encoded in the genome of this human isolate. The genome annotation revealed additional serine proteases, neuraminidase H, nitric oxide reductase, an invasion-associated protein, and acyl-CoA carboxylase subunits involved in mycolic acid biosynthesis as potential virulence factors. The cAMP-sensing transcription regulator GlxR plays a key role in controlling the expression of several genes contributing to virulence. CONCLUSION: The functional data deduced from the genome sequencing and the extended knowledge of virulence factors indicate that the human isolate C. pseudotuberculosis FRC41 is equipped with a distinct gene set promoting its survival under unfavorable environmental conditions encountered in the mammalian host.


Assuntos
Corynebacterium pseudotuberculosis/genética , Corynebacterium pseudotuberculosis/patogenicidade , Redes Reguladoras de Genes/genética , Genoma Bacteriano/genética , Linfadenite Histiocítica Necrosante/genética , Linfadenite Histiocítica Necrosante/microbiologia , Aderência Bacteriana/genética , Sequência de Bases , Criança , Corynebacterium pseudotuberculosis/isolamento & purificação , Corynebacterium pseudotuberculosis/fisiologia , Feminino , Fímbrias Bacterianas/genética , Genes Bacterianos/genética , Humanos , Ferro/metabolismo , Manganês/metabolismo , Anotação de Sequência Molecular , Regulon/genética , Análise de Sequência de DNA , Transcrição Gênica , Virulência/genética , Fatores de Virulência/genética , Fatores de Virulência/metabolismo , Zinco/metabolismo
18.
Emerg Microbes Infect ; 8(1): 17-28, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30866756

RESUMO

An outbreak with a remarkable Listeria monocytogenes clone causing 163 cases of non-invasive listeriosis occurred in Germany in 2015. Core genome multi locus sequence typing grouped non-invasive outbreak isolates and isolates obtained from related food samples into a single cluster, but clearly separated genetically close isolates obtained from invasive listeriosis cases. A comparative genomic approach identified a premature stop codon in the chiB gene, encoding one of the two L. monocytogenes chitinases, which clustered with disease outcome. Correction of this premature stop codon in one representative gastroenteritis outbreak isolate restored chitinase production, but effects in infection experiments were not found. While the exact role of chitinases in virulence of L. monocytogenes is still not fully understood, our results now clearly show that ChiB-derived activity is not required to establish L. monocytogenes gastroenteritis in humans. This limits a possible role of ChiB in human listeriosis to later steps of the infection.


Assuntos
Quitinases/genética , Surtos de Doenças , Gastroenterite/microbiologia , Listeria monocytogenes/classificação , Listeria monocytogenes/isolamento & purificação , Listeriose/epidemiologia , Adolescente , Adulto , Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana , Células CACO-2 , Criança , Pré-Escolar , Códon de Terminação , Feminino , Microbiologia de Alimentos , Gastroenterite/epidemiologia , Genômica , Alemanha/epidemiologia , Células HeLa , Células Hep G2 , Humanos , Lactente , Listeria monocytogenes/enzimologia , Listeria monocytogenes/patogenicidade , Masculino , Pessoa de Meia-Idade , Tipagem de Sequências Multilocus , Filogenia , Fatores de Virulência/genética , Adulto Jovem
19.
J Biotechnol ; 136(1-2): 11-21, 2008 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-18367281

RESUMO

Corynebacterium urealyticum is a lipid-requiring, urealytic bacterium of the human skin flora that has been recognized as causative agent of urinary tract infections. We report the analysis of the complete genome sequence of C. urealyticum DSM7109, which was initially recovered from a patient with alkaline-encrusted cystitis. The genome sequence was determined by a combination of pyrosequencing and Sanger technology. The chromosome of C. urealyticum DSM7109 has a size of 2,369,219bp and contains 2024 predicted coding sequences, of which 78% were considered as orthologous with genes in the Corynebacterium jeikeium K411 genome. Metabolic analysis of the lipid-requiring phenotype revealed the absence of a fatty acid synthase gene and the presence of a beta-oxidation pathway along with a large repertoire of auxillary genes for the degradation of exogenous fatty acids. A urease locus with the gene order ureABCEFGD may play a pivotal role in virulence of C. urealyticum by the alkalinization of human urine and the formation of struvite stones. Multidrug resistance of C. urealyticum DSM7109 is mediated by transposable elements, conferring resistances to macrolides, lincosamides, ketolides, aminoglycosides, chloramphenicol, and tetracycline. The complete genome sequence of C. urealyticum revealed a detailed picture of the lifestyle of this opportunistic human pathogen.


Assuntos
Proteínas de Bactérias/genética , Mapeamento Cromossômico/métodos , Corynebacterium/genética , Genoma Bacteriano/genética , Fases de Leitura Aberta/genética , Análise de Sequência de DNA/métodos , Sequência de Bases , Dados de Sequência Molecular
20.
Genome Announc ; 6(2)2018 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-29326206

RESUMO

Escherichia coli can colonize the urinary bladder without causing a disease response in the host. This asymptomatic bacteriuria (ABU) can protect against recurrent symptomatic urinary tract infection by virulent bacteria. Here, we report the whole-genome sequences of nine E. coli ABU isolates from diabetic patients.

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