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1.
BMC Infect Dis ; 24(1): 988, 2024 Sep 17.
Article in English | MEDLINE | ID: mdl-39289626

ABSTRACT

BACKGROUND: Corynebacterium striatum (C. striatum), a common skin and mucosal colonizer, is increasingly considered as an opportunistic pathogen causing bloodstream infections (BSIs). This study aims to investigate the clinical features and outcomes of C. striatum-BSI. METHODS: We included hospitalized cases with C. striatum-positive blood cultures from January 2014 to June 2022 and classified them into C. striatum-BSI group and contamination group; Clinical characteristics, treatments, and outcomes were compared between the C. striatum-BSI group and contamination group, Methicillin-resistant Staphylococcus aureus (MRSA)-BSI and Methicillin-resistant Staphylococcus epidermidis (MRSE)-BSI. RESULTS: Fifty-three patients with positive C. striatum blood cultures were identified. Among them, 25 patients were classified as C. striatum-BSI, with 21 as contamination cases. And 62 cases of MRSA-BSI and 44 cases of MRSE-BSI were identified. Compared to the contaminated group, the C. striatum-BSI group had a shorter time to positivity of blood cultures (27.0 h vs. 42.5 h, P = 0.011). C. striatum-BSI group had a longer time to positivity (27 h) when compared to both the MRSA (20 h) and MRSE groups (19 h) (p < 0.05). Appropriate therapy within 24 h of BSI onset was significantly lower in the C. striatum group (28%) compared to the MRSA (64.5%) and MRSE (65.9%) groups (p < 0.005). The 28-day mortality was higher in the C. striatum group (52.0%) compared to the MRSA (25.8%) and MRSE (18.2%) groups.  CONCLUSIONS: Given the distinct characteristics of C. striatum-BSI, including a longer time to positivity than other Gram-positive bacteria and higher mortality rates, we suggest prescribing early appropriate antibiotics if C. striatum-BSI is suspected.


Subject(s)
Bacteremia , Corynebacterium Infections , Corynebacterium , Methicillin-Resistant Staphylococcus aureus , Humans , Corynebacterium/isolation & purification , Corynebacterium/classification , Corynebacterium/genetics , Male , Female , Middle Aged , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Methicillin-Resistant Staphylococcus aureus/drug effects , Corynebacterium Infections/microbiology , Corynebacterium Infections/drug therapy , Bacteremia/microbiology , Bacteremia/drug therapy , Bacteremia/mortality , Aged , Staphylococcus epidermidis/isolation & purification , Staphylococcal Infections/microbiology , Staphylococcal Infections/mortality , Staphylococcal Infections/drug therapy , Adult , Anti-Bacterial Agents/therapeutic use , Anti-Bacterial Agents/pharmacology , Retrospective Studies , Aged, 80 and over
2.
Future Microbiol ; 19(11): 963-970, 2024.
Article in English | MEDLINE | ID: mdl-39109653

ABSTRACT

Non-diphtheroid Corynebacterium sepsis is rare and has affected only immunocompromised or particularly predisposed patients so far. We present the first case of urosepsis caused by Corynebacterium aurimucosum in a 67-year-old woman, without any known immunodeficiencies and in absence of any immunosuppressive therapy, admitted to the hospital for fever and acute dyspnea. This work suggests a new approach in evaluating the isolation of Corynebacteria, especially if isolated from blood. In particular, it highlights the potential infectious role of C. aurimucosum (often considered a contaminant and only rarely identified as an etiological agent of infections) and its clinical consequences, detailing also interesting aspects about its microbiological diagnosis and relative therapy and clarifying contrasting data of literature.


[Box: see text].


Subject(s)
Corynebacterium Infections , Corynebacterium , Sepsis , Urinary Tract Infections , Humans , Corynebacterium/isolation & purification , Corynebacterium/genetics , Corynebacterium/pathogenicity , Corynebacterium/classification , Aged , Female , Corynebacterium Infections/microbiology , Corynebacterium Infections/diagnosis , Corynebacterium Infections/drug therapy , Sepsis/microbiology , Sepsis/drug therapy , Urinary Tract Infections/microbiology , Urinary Tract Infections/drug therapy , Urinary Tract Infections/diagnosis , Anti-Bacterial Agents/therapeutic use
3.
Res Vet Sci ; 179: 105396, 2024 Nov.
Article in English | MEDLINE | ID: mdl-39213744

ABSTRACT

Although diphtheria is a vaccine-preventable disease, numerous cases are still reported around the world, as well as outbreaks in countries, including European ones. Species of the Corynebacterium diphtheriae complex are potentially toxigenic and, therefore, must be considered given the possible consequences, such as the circulation of clones and transmission of antimicrobial resistance and virulence genes. Recently, Corynebacterium rouxii was characterized and included among the valid species of the complex. Therefore, two cases of C. rouxii infection arising from infections in domestic animals are presented here. We provide molecular characterization, phylogenetic analyses, genome sequencing, and CRISPR-Cas analyses to contribute to a better understanding of the molecular bases, pathogenesis, and epidemiological monitoring of this species, which is still little studied. We confirmed its taxonomic position with genome sequencing and in silico analysis and identified the ST-918 for both strains. The clinical isolates were sensitive resistance to benzylpenicillin and rifampin. Antimicrobial resistance genes, including tetB, rpoB2, and rbpA genes, were predicted. The bla and ampC genes were not found. Several virulence factors were also detected, including adhesion, iron uptake systems, gene regulation (dtxR), and post-translational modification (MdbA). Finally, one prophage and the Type I-E CRISPR-Cas system were identified.


Subject(s)
Anti-Bacterial Agents , Corynebacterium Infections , Corynebacterium , Dog Diseases , Phylogeny , Rifampin , Animals , Corynebacterium/genetics , Corynebacterium/drug effects , Dog Diseases/microbiology , Dogs , Rifampin/pharmacology , Corynebacterium Infections/veterinary , Corynebacterium Infections/microbiology , Anti-Bacterial Agents/pharmacology , Genome, Bacterial , Drug Resistance, Bacterial/genetics , Penicillins/pharmacology
4.
J Agric Food Chem ; 72(30): 16877-16888, 2024 Jul 31.
Article in English | MEDLINE | ID: mdl-39038232

ABSTRACT

Esterases are crucial for aryloxyphenoxypropionate herbicide (AOPP) biodegradation. However, the underlying molecular mechanisms of AOPP biodegradation by esterases are poorly understood. In the current work, Corynebacterium sp. Z-1 was isolated and found to degrade multiple AOPPs, including quizalofop-p-ethyl (QPE), haloxyfop-p-methyl (HPM), fenoxaprop-p-ethyl (FPE), cyhalofop-butyl (CYB), and clodinafop-propargyl (CFP). A novel esterase, QfeH, which catalyzes the cleavage of ester bonds in AOPPs to form AOPP acids, was identified from strain Z-1. The catalytic activities of QfeH toward AOPPs decreased in the following order: CFP > FPE > CYB > QPE > HPM. Molecular docking, computational analyses, and site-directed mutagenesis indicated the catalytic mechanisms of QfeH-mediated degradation of different AOPPs. Notably, the key residue S159 is essential for the activity of QfeH. Moreover, V222Y, T227M, T227A, A271R, and M275K mutants, exhibiting 2.9-5.0 times greater activity than QfeH, were constructed. This study facilitates the mechanistic understanding of AOPPs bioremediation by esterases.


Subject(s)
Biodegradation, Environmental , Corynebacterium , Esterases , Herbicides , Herbicides/metabolism , Herbicides/chemistry , Esterases/metabolism , Esterases/genetics , Esterases/chemistry , Corynebacterium/metabolism , Corynebacterium/genetics , Corynebacterium/enzymology , Bacterial Proteins/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/chemistry , Molecular Docking Simulation , Propionates/metabolism
5.
Cancer Sci ; 115(8): 2738-2750, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38888048

ABSTRACT

Pancreatic head cancer (PHC) and pancreatic body/tail cancer (PBTC) have distinct clinical and biological behaviors. The microbial and metabolic differences in PHC and PBTC have not been studied. The pancreatic microbiota and metabolome of 15 PHC and 8 PBTC tissues and their matched nontumor tissues were characterized using 16S rRNA amplicon sequencing and untargeted metabolomics. At the genus level, Bradyrhizobium was increased while Corynebacterium and Ruminococcus were decreased in the PHC tissues (Head T) compared with the matched nontumor tissues (Head N) significantly. Shuttleworthia, Bacillus, and Bifidobacterium were significantly decreased in the PBTC tissues (Body/Tail T) compared with the matched nontumor tissues (Body/Tail N). Significantly, Ileibacterium was increased whereas Pseudoxanthomonas was decreased in Head T and Body/Tail T, and Lactobacillus was increased in Head T but decreased in Body/Tail T. A total of 102 discriminative metabolites were identified between Head T and Head N, which were scattered through linoleic acid metabolism and purine metabolism pathways. However, there were only four discriminative metabolites between Body/Tail T and Body/Tail N, which were related to glycerophospholipid metabolism and autophagy pathways. The differential metabolites in PHC and PBTC were commonly enriched in alpha-linolenic acid metabolism and choline metabolism in cancer pathways. Eubacterium decreased in Head T was positively correlated with decreased linoleic acid while negatively correlated with increased arachidyl carnitine and stearoylcarnitine. Bacillus decreased in Body/Tail T was negatively correlated with increased L-carnitine. These microbiota and metabolites deserve further investigations to reveal their roles in the pathogenesis of PHC and PBTC, providing clues for future treatments.


Subject(s)
Pancreatic Neoplasms , RNA, Ribosomal, 16S , Humans , Pancreatic Neoplasms/microbiology , Pancreatic Neoplasms/metabolism , Male , Middle Aged , Female , Aged , RNA, Ribosomal, 16S/genetics , Metabolome , Microbiota , Metabolomics/methods , Pancreas/metabolism , Pancreas/microbiology , Corynebacterium/metabolism , Corynebacterium/genetics
6.
Toxins (Basel) ; 16(6)2024 May 26.
Article in English | MEDLINE | ID: mdl-38922140

ABSTRACT

Diphtheria toxin (DT) is the main virulence factor of Corynebacterium diphtheriae, C. ulcerans and C. pseudotuberculosis. Moreover, new Corynebacterium species with the potential to produce diphtheria toxin have also been described. Therefore, the detection of the toxin is the most important test in the microbiological diagnosis of diphtheria and other corynebacteria infections. Since the first demonstration in 1888 that DT is a major virulence factor of C. diphtheriae, responsible for the systemic manifestation of the disease, various methods for DT detection have been developed, but the diagnostic usefulness of most of them has not been confirmed on a sufficiently large group of samples. Despite substantial progress in the science and diagnostics of infectious diseases, the Elek test is still the basic recommended diagnostic test for DT detection. The challenge here is the poor availability of an antitoxin and declining experience even in reference laboratories due to the low prevalence of diphtheria in developed countries. However, recent and very promising assays have been developed with the potential for use as rapid point-of-care testing (POCT), such as ICS and LFIA for toxin detection, LAMP for tox gene detection, and biosensors for both.


Subject(s)
Diphtheria Toxin , Diphtheria , Diphtheria Toxin/genetics , Humans , Diphtheria/diagnosis , Diphtheria/microbiology , Corynebacterium/genetics , Corynebacterium diphtheriae
7.
Microbiol Spectr ; 12(7): e0344123, 2024 Jul 02.
Article in English | MEDLINE | ID: mdl-38864649

ABSTRACT

This study aimed to characterize the composition of intestinal and nasal microbiota in septic patients and identify potential microbial biomarkers for diagnosis. A total of 157 subjects, including 89 with sepsis, were enrolled from the affiliated hospital. Nasal swabs and fecal specimens were collected from septic and non-septic patients in the intensive care unit (ICU) and Department of Respiratory and Critical Care Medicine. DNA was extracted, and the V4 region of the 16S rRNA gene was amplified and sequenced using Illumina technology. Bioinformatics analysis, statistical processing, and machine learning techniques were employed to differentiate between septic and non-septic patients. The nasal microbiota of septic patients exhibited significantly lower community richness (P = 0.002) and distinct compositions (P = 0.001) compared to non-septic patients. Corynebacterium, Staphylococcus, Acinetobacter, and Pseudomonas were identified as enriched genera in the nasal microbiota of septic patients. The constructed machine learning model achieved an area under the curve (AUC) of 89.08, indicating its efficacy in differentiating septic and non-septic patients. Importantly, model validation demonstrated the effectiveness of the nasal microecological diagnosis prediction model with an AUC of 84.79, while the gut microecological diagnosis prediction model had poor predictive performance (AUC = 49.24). The nasal microbiota of ICU patients effectively distinguishes sepsis from non-septic cases and outperforms the gut microbiota. These findings have implications for the development of diagnostic strategies and advancements in critical care medicine.IMPORTANCEThe important clinical significance of this study is that it compared the intestinal and nasal microbiota of sepsis with non-sepsis patients and determined that the nasal microbiota is more effective than the intestinal microbiota in distinguishing patients with sepsis from those without sepsis, based on the difference in the lines of nasal specimens collected.


Subject(s)
Bacteria , Biomarkers , Feces , Intensive Care Units , Microbiota , RNA, Ribosomal, 16S , Sepsis , Humans , Sepsis/diagnosis , Sepsis/microbiology , Male , Female , Middle Aged , Aged , RNA, Ribosomal, 16S/genetics , Biomarkers/analysis , Bacteria/isolation & purification , Bacteria/genetics , Bacteria/classification , Feces/microbiology , Adult , Machine Learning , Gastrointestinal Microbiome , Nose/microbiology , Corynebacterium/isolation & purification , Corynebacterium/genetics , Acinetobacter/isolation & purification , Acinetobacter/genetics , Aged, 80 and over , Staphylococcus/isolation & purification , Staphylococcus/genetics , Pseudomonas/isolation & purification , Pseudomonas/genetics
8.
J Glob Antimicrob Resist ; 38: 181-186, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38936471

ABSTRACT

Herein, we combined different bioinformatics tools and databases (BV-BRC, ResFinder, RAST, and KmerResistance) to perform a prediction of antimicrobial resistance (AMR) in the genomic sequences of 107 Corynebacterium striatum isolates for which trustable antimicrobial susceptibility (AST) phenotypes could be retrieved. Then, the reliabilities of the AMR predictions were evaluated by different metrics: area under the ROC curve (AUC); Major Error Rates (MERs) and Very Major Error Rates (VMERs); Matthews Correlation Coefficient (MCC); F1-Score; and Accuracy. Out of 15 genes that were reliably detected in the C. striatum isolates, only tetW yielded predictive values for tetracycline resistance that were acceptable considering Food and Drug Administration (FDA)'s criteria for quality (MER < 3.0% and VMER with a 95% C.I. ≤1.5-≤7.5); this was accompanied by a MCC score higher than 0.9 for this gene. Noteworthy, our results indicate that other commonly used metrics (AUC, F1-score, and Accuracy) may render overoptimistic evaluations of AMR-prediction reliabilities on imbalanced datasets. Accordingly, out of 10 genes tested by PCR on additional multidrug-resistant Corynebacterium spp. isolates (n = 18), the tetW gene rendered the best agreement values with AST profiles (94.11%). Overall, our results indicate that genome-based AMR prediction can still be challenging for MDR clinical isolates of emerging Corynebacterium spp.


Subject(s)
Anti-Bacterial Agents , Computational Biology , Corynebacterium , Drug Resistance, Multiple, Bacterial , Genome, Bacterial , Microbial Sensitivity Tests , Whole Genome Sequencing , Corynebacterium/genetics , Corynebacterium/drug effects , Corynebacterium/isolation & purification , Corynebacterium/classification , Drug Resistance, Multiple, Bacterial/genetics , Anti-Bacterial Agents/pharmacology , Humans , Corynebacterium Infections/microbiology
9.
Fungal Genet Biol ; 173: 103898, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38815692

ABSTRACT

The skin and its microbiome function to protect the host from pathogen colonization and environmental stressors. In this study, using the Wisconsin Miniature Swine™ model, we characterize the porcine skin fungal and bacterial microbiomes, identify bacterial isolates displaying antifungal activity, and use whole-genome sequencing to identify biosynthetic gene clusters encoding for secondary metabolites that may be responsible for the antagonistic effects on fungi. Through this comprehensive approach of paired microbiome sequencing with culturomics, we report the discovery of novel species of Corynebacterium and Rothia. Further, this study represents the first comprehensive evaluation of the porcine skin mycobiome and the evaluation of bacterial-fungal interactions on this surface. Several diverse bacterial isolates exhibit potent antifungal properties against opportunistic fungal pathogens in vitro. Genomic analysis of inhibitory species revealed a diverse repertoire of uncharacterized biosynthetic gene clusters suggesting a reservoir of novel chemical and biological diversity. Collectively, the porcine skin microbiome represents a potential unique source of novel antifungals.


Subject(s)
Fungi , Microbiota , Skin , Animals , Skin/microbiology , Swine/microbiology , Microbiota/genetics , Fungi/genetics , Fungi/drug effects , Antifungal Agents/pharmacology , Antibiosis , Mycobiome/genetics , Bacteria/genetics , Bacteria/classification , Bacteria/drug effects , Bacteria/isolation & purification , Bacteria/metabolism , Corynebacterium/genetics , Corynebacterium/drug effects , Swine, Miniature/microbiology , Multigene Family , Whole Genome Sequencing , Secondary Metabolism/genetics
10.
Eur J Clin Microbiol Infect Dis ; 43(7): 1495-1501, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38801486

ABSTRACT

Corynebacterium striatum is an emerging nosocomial pathogen. This is the first report showing the presence of three distinct multidrug resistant lineages of C. striatum among patients in a UK hospital. The presence of ErmX, Tet(W), Bla and AmpC proteins, and mutations in gyrA gene are associated with the resistance to clindamycin, doxycycline, penicillin and moxifloxacin, respectively. These strains are equipped with several corynebacterial virulence genes including two SpaDEF-type and a novel pilus gene cluster, which needs further molecular characterisation. This study highlights a need of developing an active surveillance strategy for routine monitoring and preventing potential cross-transmission among susceptible patients.


Subject(s)
Anti-Bacterial Agents , Corynebacterium Infections , Corynebacterium , Drug Resistance, Multiple, Bacterial , Phylogeny , Tertiary Care Centers , Humans , Corynebacterium/genetics , Corynebacterium/drug effects , Corynebacterium/isolation & purification , Corynebacterium/classification , Drug Resistance, Multiple, Bacterial/genetics , Corynebacterium Infections/microbiology , Corynebacterium Infections/epidemiology , United Kingdom/epidemiology , Anti-Bacterial Agents/pharmacology , Microbial Sensitivity Tests , Cross Infection/microbiology , Cross Infection/epidemiology , Male , Virulence Factors/genetics , Bacterial Proteins/genetics , Female
11.
Front Cell Infect Microbiol ; 14: 1361045, 2024.
Article in English | MEDLINE | ID: mdl-38572320

ABSTRACT

Introduction: Over the past decade, Corynebacterium striatum (C. striatum), an emerging multidrug-resistant (MDR) pathogen, has significantly challenged healthcare settings, especially those involving individuals with weakened immune systems. The rise of these superbugs necessitates innovative solutions. Methods: This study aimed to isolate and characterize bacteriophages targeting MDR-C. striatum. Utilizing 54 MDR-C. striatum isolates from a local hospital as target strains, samples were collected from restroom puddles for phage screening. Dot Plaque and Double-layer plate Assays were employed for screening. Results: A novel temperate bacteriophage, named CSP1, was identified through a series of procedures, including purification, genome extraction, sequencing, and one-step growth curves. CSP1 possesses a 39,752 base pair circular double-stranded DNA genome with HK97-like structural proteins and potential for site-specific recombination. It represents a new species within the unclassified Caudoviricetes class, as supported by transmission electron microscopy, genomic evolutionary analysis, and collinearity studies. Notably, CSP1 infected and lysed 21 clinical MDR-C. striatum isolates, demonstrating a wide host range. The phage remained stable in conditions ranging from -40 to 55°C, pH 4 to 12, and in 0.9% NaCl buffer, showing no cytotoxicity. Discussion: The identification of CSP1 as the first phage targeting clinical C. striatum strains opens new possibilities in bacteriophage therapy research, and the development of diagnostic and therapeutic tools against pathogenic bacteria.


Subject(s)
Bacteriophages , Corynebacterium Infections , Humans , Bacteriophages/genetics , Corynebacterium/genetics , Corynebacterium Infections/microbiology , Genomics , Anti-Bacterial Agents
12.
Braz J Microbiol ; 55(2): 1405-1414, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38598149

ABSTRACT

BACKGROUND: Corynebacterium spp. are widely disseminated in the environment, and they are part of the skin and mucosal microbiota of animals and humans. Reports of human infections by Corynebacterium spp. have increased considerably in recent years and the appearance of multidrug resistant isolates around the world has drawn attention. OBJECTIVES: To describe a new species of Corynebacterium from human tissue bone is described after being misidentified using available methods. METHODS: For taxonomic analyses, phylogenetic analysis of 16S rRNA and rpoB genes, in silico DNA-DNA hybridization, average nucleotide and amino acid identity, multilocus sequence analysis, and phylogenetic analysis based on the complete genome were used. FINDINGS: Genomic taxonomic analyzes revealed values of in silico DNA-DNA hybridization, average nucleotide and amino acids identity below the values necessary for species characterization between the analyzed isolates and the closest phylogenetic relative Corynebacterium aurimucosum DSM 44532T. MAIN CONCLUSIONS: Genomic taxonomic analyzes indicate that the isolates analyzed comprise a new species of the Corynebacterium genus, which we propose to name Corynebacterium hiratae sp. nov. with isolate 332T (= CBAS 826T = CCBH 35,014T) as the type strain.


Subject(s)
Corynebacterium Infections , Corynebacterium , DNA, Bacterial , Phylogeny , RNA, Ribosomal, 16S , Corynebacterium/genetics , Corynebacterium/classification , Corynebacterium/isolation & purification , Humans , RNA, Ribosomal, 16S/genetics , DNA, Bacterial/genetics , Corynebacterium Infections/microbiology , Bone and Bones/microbiology , Multilocus Sequence Typing , Genome, Bacterial , Bacterial Typing Techniques , Nucleic Acid Hybridization
13.
Clin Lab ; 70(3)2024 Mar 01.
Article in English | MEDLINE | ID: mdl-38469769

ABSTRACT

BACKGROUND: The goal was to report a rare case of lymphadenitis caused by Corynebacterium tuberculostearicum, and the laboratory's coping approach in the isolation and identification of this rare pathogen to improve the understanding of the disease. METHODS: Lymph node biopsy was performed in a patient with suspected tuberculous lymphadenitis, and the biopsy tissue was isolated and cultured. RESULTS: The culture was Gram positive Corynebacterium, which was identified as Corynebacterium tuberculostearicum by microbial mass spectrometry and 16S rRNA gene sequencing. Antimicrobial susceptibility test showed that the drug was sensitive to daptomycin, doxycycline, gentamicin, linezolid, vancomycin, and meropenem, but resistant to ciprofloxacin, clindamycin, erythromycin, rifampicin, compound sulfamethoxazole, ceftriaxone, and cefepime. CONCLUSIONS: This is a case of Corynebacterium tuberculostearicum infection. Case reports of Corynebacterium tuberculostearicum infection are relatively rare in China. Through case study, we can provide help for laboratory isolation, identification, clinical diagnosis, and treatment.


Subject(s)
Corynebacterium Infections , Corynebacterium , Humans , RNA, Ribosomal, 16S/genetics , Corynebacterium/genetics , Corynebacterium Infections/diagnosis , Corynebacterium Infections/drug therapy , Corynebacterium Infections/microbiology , Vancomycin/therapeutic use , Anti-Bacterial Agents/therapeutic use , Microbial Sensitivity Tests
14.
Appl Microbiol Biotechnol ; 108(1): 251, 2024 Mar 04.
Article in English | MEDLINE | ID: mdl-38436751

ABSTRACT

Product secretion from an engineered cell can be advantageous for microbial cell factories. Extensive work on nucleotide manufacturing, one of the most successful microbial fermentation processes, has enabled Corynebacterium stationis to transport nucleotides outside the cell by random mutagenesis; however, the underlying mechanism has not been elucidated, hindering its applications in transporter engineering. Herein, we report the nucleotide-exporting major facilitator superfamily (MFS) transporter from the C. stationis genome and its hyperactive mutation at the G64 residue. Structural estimation and molecular dynamics simulations suggested that the activity of this transporter improved via two mechanisms: (1) enhancing interactions between transmembrane helices through the conserved "RxxQG" motif along with substrate binding and (2) trapping substrate-interacting residue for easier release from the cavity. Our results provide novel insights into how MFS transporters change their conformation from inward- to outward-facing states upon substrate binding to facilitate efflux and can contribute to the development of rational design approaches for efflux improvements in microbial cell factories. KEYPOINTS: • An MFS transporter from C. stationis genome and its mutation at residue G64 were assessed • It enhanced the transporter activity by strengthening transmembrane helix interactions and trapped substrate-interacting residues • Our results contribute to rational design approach development for efflux improvement.


Subject(s)
Corynebacterium , Membrane Transport Proteins , Membrane Transport Proteins/genetics , Biological Transport , Corynebacterium/genetics , Nucleotides
15.
J Antimicrob Chemother ; 79(5): 1023-1029, 2024 05 02.
Article in English | MEDLINE | ID: mdl-38497972

ABSTRACT

BACKGROUND: Corynebacterium (C.) sp. 22KM0430 related to C. oculi and isolated from a dog exhibited resistance to tetracycline, and its WGS analysis revealed a putative resistance gene on a 35 562-bp plasmid also harbouring the MLSB resistance gene erm(X). OBJECTIVES: To characterize the novel tetracycline resistance gene tet(65) and demonstrate its functionality by expression in C. glutamicum and Escherichia coli and plasmid curing of the host strain. METHODS: tet(65) was cloned with and without its repressor tetR(65) and expressed in C. glutamicum DSM20300 and E. coli DH5α. Plasmid was cured by non-selective passages. Minimal inhibitory concentrations (MICs) of tetracyclines were determined according to CLSI guidelines. Association of tet(65) with efflux was shown by the addition of reserpine to MIC assays. Phylogenetic position and transmembrane structure of Tet(65) were analysed using MEGA11 and DeepTMHMM. RESULTS: Tet(65) shows 73% amino acid identity with the closest related Tet(Z), contains 12 transmembrane domains and is structurally related to the Major Facilitator Superfamily. The tetracycline MICs decreased in the plasmid-cured strain and increased when tet(65) was expressed in C. glutamicum and in E. coli. The MICs of tetracycline decreased in the presence of reserpine indicating that tet(65) functions as an efflux pump. A GenBank search also identified tet(65) in C. diphtheriae and Brevibacterium (B.) casei and B. luteolum. CONCLUSIONS: A novel tetracycline efflux gene tet(65) was identified in a C. oculi related species and was also present in the human pathogen C. diphtheriae and in Brevibacterium species indicating broader potential for dissemination.


Subject(s)
Anti-Bacterial Agents , Corynebacterium , Escherichia coli , Microbial Sensitivity Tests , Plasmids , Tetracycline Resistance , Plasmids/genetics , Tetracycline Resistance/genetics , Anti-Bacterial Agents/pharmacology , Escherichia coli/genetics , Escherichia coli/drug effects , Corynebacterium/genetics , Corynebacterium/drug effects , Animals , Drug Resistance, Multiple, Bacterial/genetics , Phylogeny , Dogs , Tetracycline/pharmacology , Cloning, Molecular , Corynebacterium Infections/microbiology , Dog Diseases/microbiology
16.
BMC Microbiol ; 24(1): 14, 2024 Jan 05.
Article in English | MEDLINE | ID: mdl-38178003

ABSTRACT

BACKGROUND: Reliable species identification of cultured isolates is essential in clinical bacteriology. We established a new study algorithm named NOVA - Novel Organism Verification and Analysis to systematically analyze bacterial isolates that cannot be characterized by conventional identification procedures MALDI-TOF MS and partial 16 S rRNA gene sequencing using Whole Genome Sequencing (WGS). RESULTS: We identified a total of 35 bacterial strains that represent potentially novel species. Corynebacterium sp. (n = 6) and Schaalia sp. (n = 5) were the predominant genera. Two strains each were identified within the genera Anaerococcus, Clostridium, Desulfovibrio, and Peptoniphilus, and one new species was detected within Citrobacter, Dermabacter, Helcococcus, Lancefieldella, Neisseria, Ochrobactrum (Brucella), Paenibacillus, Pantoea, Porphyromonas, Pseudoclavibacter, Pseudomonas, Psychrobacter, Pusillimonas, Rothia, Sneathia, and Tessaracoccus. Twenty-seven of 35 strains were isolated from deep tissue specimens or blood cultures. Seven out of 35 isolated strains identified were clinically relevant. In addition, 26 bacterial strains that could only be identified at the species level using WGS analysis, were mainly organisms that have been identified/classified very recently. CONCLUSION: Our new algorithm proved to be a powerful tool for detection and identification of novel bacterial organisms. Publicly available clinical and genomic data may help to better understand their clinical and ecological role. Our identification of 35 novel strains, 7 of which appear to be clinically relevant, shows the wide range of undescribed pathogens yet to define.


Subject(s)
Bacteria , Corynebacterium , Bacteria/genetics , Whole Genome Sequencing , Corynebacterium/genetics , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , RNA, Ribosomal, 16S/genetics , Bacterial Typing Techniques/methods
17.
Article in English | MEDLINE | ID: mdl-38289223

ABSTRACT

The taxonomic status of strain P5891T, isolated from an Adélie penguin beak swab, was investigated. Based on the 16S rRNA gene sequence, the strain was identified as a potentially novel Corynebacterium species, with the highest sequence similarities to Corynebacterium rouxii FRC0190T (96.7 %) and Corynebacterium epidermidicanis DSM 45586T (96.6 %). The average nucleotide identity values between strain P5891T and C. rouxii FRC0190T and C. epidermidicanis DSM 45586T were 68.2 and 69.2 %, respectively. The digital DNA-DNA hybridization values between strain P5891T and C. rouxii FRC0190T and C. epidermidicanis DSM 45586T were 23.7 and 21.4 %, respectively. Phylogenetic trees based on the 16S rRNA sequence placed strain P5891T in a separate branch with Corynebacterium canis 1170T and Corynebacterium freiburgense 1045T, while a phylogenomic tree based on the Corynebacterium species core genome placed the strain next to Corynebacterium choanae 200CHT. Extensive phenotyping and genomic analyses clearly confirmed that strain P5891T represents a novel species of the genus Corynebacterium, for which the name Corynebacterium mendelii sp. nov. is proposed, with the type strain P5891T (=CCM 8862T=LMG 31627T).


Subject(s)
Spheniscidae , Animals , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , DNA, Bacterial/genetics , Bacterial Typing Techniques , Base Composition , Fatty Acids/chemistry , Bacteria , Corynebacterium/genetics , Mouth
18.
J Appl Microbiol ; 135(1)2024 Jan 02.
Article in English | MEDLINE | ID: mdl-38130215

ABSTRACT

AIMS: This study aimed to characterize the first complete genome of Corynebacterium parakroppenstedtii and clarify the evolutionary relationship in the Corynebacterium kroppenstedtii complex (CKC) by using comparative genomics analysis. METHODS AND RESULTS: The genome of isolate yu01 from a breast specimen was sequenced, and 35 CKC genomes were collected. Analysis of 16S rRNA, rpoB, and fusA suggested ambiguous identification, whereas ANI analysis assigned isolate yu01 as Coryne. parakroppenstedtii. The fourth genospecies "Corynebacterium aliikroppenstedtii" was identified in CKC. Comparative genomics analysis suggested that the genomic arrangement in CKC was highly conserved. A total of 43 potential virulence genes and 79 species-specific genes were detected. Most genome-based phylogenetic analysis were incapable of resolving the interspecific evolutionary relationships among CKCs. A total of 20 core genes were found to be distinguishable in CKC. CONCLUSIONS: This study suggested the limited divergence and unavailability of normal single gene-based identification in CKC and questioned the precise species of strains associated with mastitis, identified as Coryne. kroppenstedtii in previous studies. The 20 genes showed potential to enhance the methods for the identification and epidemiological investigation of CKC.


Subject(s)
Corynebacterium Infections , Mastitis , Female , Humans , Corynebacterium Infections/complications , Corynebacterium Infections/microbiology , Phylogeny , RNA, Ribosomal, 16S/genetics , Corynebacterium/genetics , Mastitis/complications , Genomics
19.
BMC Microbiol ; 23(1): 385, 2023 Dec 06.
Article in English | MEDLINE | ID: mdl-38053056

ABSTRACT

BACKGROUND: Corynebacterium pyruviciproducens is a recently described species of Corynebacterium. There are few reports on the microbiological characteristics of the new species, and there is a lack of reports on the genomic analysis of the species. RESULTS: This study involved a clinical isolate from the pus of a hospital patient with sebaceous gland abscesses. The clinically isolated strain was identified as C. pyruviciproducens strain WYJY-01. In this study, referring to Koch's postulates, we observed the pathological changes of animal models infected by intraperitoneal injection and subcutaneous injection of pure culture of the strain WYJY-01. Furthermore, the strain WYJY-01 was isolated and cultured again from animal models' subcutaneous abscess drainage fluid. Subsequently, the genomics of the strain WYJY-01 was analyzed. By comparing various gene databases, this study predicted the core secondary metabolite gene cluster of the strain WYJY-01, virulence factor genes carried by prophage, pathogenicity islands, and resistance islands. In addition, the genomes of C. pyruviciproducens strain WYJY-01, ATCC BAA-1742 T, and UMB0763 were analyzed by comparative genomics, and the differential genes of strain WYJY-01 were compared, and their functions were analyzed. CONCLUSION: The findings showed that the strain WYJY-01 had pathogenicity, supplementing the phenotype characteristics of C. pyruviciproducens. Meanwhile, this research revealed the possible molecular mechanism of the pathogenicity of the strain WYJY-01 at the gene level through whole genome sequence analysis, providing a molecular basis for further research.


Subject(s)
Corynebacterium , Genomics , Animals , Humans , Corynebacterium/genetics , Virulence/genetics , Phenotype , Genome, Bacterial , Phylogeny
20.
BMC Vet Res ; 19(1): 251, 2023 Nov 29.
Article in English | MEDLINE | ID: mdl-38031130

ABSTRACT

BACKGROUND: The genus Corynebacterium comprises well-known animal and human pathogens as well as commensals of skin and mucous membranes. Species formerly regarded as contaminants are increasingly being recognized as opportunistic pathogens. Corynebacterium oculi has recently been described as a human ocular pathogen but has so far not been reported in dogs. CASE PRESENTATION: Here we present two cases of infection with a novel Corynebacterium sp., a corneal ulcer and a case of bacteriuria. The two bacterial isolates could not be identified by MALDI-TOF MS. While 16 S rRNA gene (99.3% similarity) and rpoB (96.6% identity) sequencing led to the preliminary identification of the isolates as Corynebacterium (C.) oculi, whole genome sequencing revealed the strains to be closely related to, but in a separate cluster from C. oculi. Antimicrobial susceptibility testing showed high minimal inhibitory concentrations of lincosamides, macrolides, tetracycline, and fluoroquinolones for one of the isolates, which also contained an erm(X) and tet-carrying plasmid as well as a nonsynonymous mutation leading to an S84I substitution in the quinolone resistance determining region of GyrA. CONCLUSIONS: While the clinical signs of both dogs were alleviated by antimicrobial treatment, the clinical significance of these isolates remains to be proven. However, considering its close relation with C. oculi, a known pathogen in humans, pathogenic potential of this species is not unlikely. Furthermore, these bacteria may act as reservoir for antimicrobial resistance genes also in a One Health context since one strain carried a multidrug resistance plasmid related to pNG3 of C. diphtheriae.


Subject(s)
Corynebacterium Infections , Dog Diseases , Animals , Dogs , Anti-Bacterial Agents/pharmacology , Corynebacterium/genetics , Corynebacterium Infections/veterinary , Corynebacterium Infections/microbiology , Drug Resistance, Bacterial/genetics , Microbial Sensitivity Tests/veterinary
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