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Corynebacterium striatum, a common constituent of the human skin microbiome, is now considered an emerging multidrug-resistant pathogen of immunocompromised and chronically ill patients. However, little is known about the molecular mechanisms in the transition from colonization to the multidrug-resistant (MDR) invasive phenotype in clinical isolates. This study performed a comprehensive pan-genomic analysis of C. striatum, including isolates from "normal skin microbiome" and from MDR infections, to gain insights into genetic factors contributing to pathogenicity and multidrug resistance in this species. For this, three novel genome sequences were obtained from clinical isolates of C. striatum of patients from Brazil, and other 24 complete or draft C. striatum genomes were retrieved from GenBank, including the ATCC6940 isolate from the Human Microbiome Project. Analysis of C. striatum strains demonstrated the presence of an open pan-genome (α = 0.852803) containing 3816 gene families, including 15 antimicrobial resistance (AMR) genes and 32 putative virulence factors. The core and accessory genomes included 1297 and 1307 genes, respectively. The identified AMR genes are primarily associated with resistance to aminoglycosides and tetracyclines. Of these, 66.6% are present in genomic islands, and four AMR genes, including aac(6')-ib7, are located in a class 1-integron. In conclusion, our data indicated that C. striatum possesses genomic characteristics favorable to the invasive phenotype, with high genomic plasticity, a robust genetic arsenal for iron acquisition, and important virulence determinants and AMR genes present in mobile genetic elements.
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Antibacterianos , Corynebacterium , Humanos , Fenótipo , Fatores de Virulência/genética , Farmacorresistência Bacteriana Múltipla/genética , Testes de Sensibilidade MicrobianaRESUMO
We have investigated the antibacterial, anti-biofilm, and anti-quorum sensing potencies of six Essential Oils (EOs) obtained from cinnamon (Cinnamomum verum), thyme (Thymus vulgaris), clove (Eugenia caryophyllata), curcuma (Curcuma longa L.), rosemary (Rosmarinus officinalis L.), and sage (Salvia officinalis). The study was conducted on 20 multidrug-resistant (MDR) S. enteritidis clinical strains. Minimum inhibitory concentrations and minimum bactericide concentrations were displayed by microdilution. The effect on biofilm formation was tested on polystyrene plates. The anti-quorum sensing effect was determined by measuring the inhibition of violacein production by Chromobacterium violaceum CV026. The influence of EOs on the adhesion of Salmonella strains to HT-29 cells was studied. The potency of S. enteritidis to infect and kill Caenorhabditis elegans was evaluated. The cinnamon, thyme, and clove EOs showed remarkable antibacterial properties. Biofilm formation was significantly reduced by the six EOs: 99.10% for cinnamon, 97.64% for clove, 95.90% for thyme, 79.84% for rosemary, 28.98% for curcuma, and 15.55% for sage. The MIC/2 of clove EO exhibited the highest percentage of inhibition of violacein production (99.03%), followed by thyme (91.68%) and cinnamon (84.13%) EOs. Thyme extracts exhibited an important anti-adhesive potency. Clove EO behaves as an effective regulator of Salmonella virulence in nematodes.
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Óleos Voláteis , Syzygium , Thymus (Planta) , Antibacterianos/farmacologia , Cinnamomum zeylanicum , Testes de Sensibilidade Microbiana , Óleos Voláteis/farmacologia , Salmonella enteritidisRESUMO
OBJECTIVES: No standard therapy, including anticoagulation regimens, is currently recommended for coronavirus disease 2019. Aim of this study was to evaluate the efficacy of anticoagulation in coronavirus disease 2019 hospitalized patients and its impact on survival. DESIGN: Multicenter international prospective registry (Health Outcome Predictive Evaluation for Corona Virus Disease 2019). SETTING: Hospitalized patients with coronavirus disease 2019. PATIENTS: Five thousand eight hundred thirty-eight consecutive coronavirus disease 2019 patients. INTERVENTIONS: Anticoagulation therapy, including prophylactic and therapeutic regimens, was obtained for each patient. MEASUREMENTS AND MAIN RESULTS: Five thousand four hundred eighty patients (94%) did not receive any anticoagulation before hospitalization. Two-thousand six-hundred one patients (44%) during hospitalization received anticoagulation therapy and it was not associated with better survival rate (81% vs 81%; p = 0.94) but with higher risk of bleeding (2.7% vs 1.8%; p = 0.03). Among patients admitted with respiratory failure (49%, n = 2,859, including 391 and 583 patients requiring invasive and noninvasive ventilation, respectively), anticoagulation started during hospitalization was associated with lower mortality rates (32% vs 42%; p < 0.01) and nonsignificant higher risk of bleeding (3.4% vs 2.7%; p = 0.3). Anticoagulation therapy was associated with lower mortality rates in patients treated with invasive ventilation (53% vs 64%; p = 0.05) without increased rates of bleeding (9% vs 8%; p = 0.88) but not in those with noninvasive ventilation (35% vs 38%; p = 0.40). At multivariate Cox' analysis mortality relative risk with anticoagulation was 0.58 (95% CI, 0.49-0.67) in patients admitted with respiratory failure, 0.50 (95% CI, 0.49-0.67) in those requiring invasive ventilation, 0.72 (95% CI, 0.51-1.01) in noninvasive ventilation. CONCLUSIONS: Anticoagulation therapy in general population with coronavirus disease 2019 was not associated with better survival rates but with higher bleeding risk. Better results were observed in patients admitted with respiratory failure and requiring invasive ventilation.
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Anticoagulantes/uso terapêutico , Tratamento Farmacológico da COVID-19 , Avaliação de Resultados em Cuidados de Saúde , Sistema de Registros , COVID-19/mortalidade , Estudos de Casos e Controles , Correlação de Dados , Comparação Transcultural , Hemorragia/induzido quimicamente , Hemorragia/mortalidade , Hospitalização , Humanos , Estudos Multicêntricos como Assunto , Estudos Prospectivos , Respiração Artificial , Insuficiência Respiratória/tratamento farmacológico , Insuficiência Respiratória/mortalidade , Risco , Taxa de Sobrevida , Resultado do TratamentoRESUMO
Corynebacterium striatum is a nosocomial pathogen which is increasingly associated with serious infections in both immunocompetent and immunocompromised patients. However, little is known about virulence factors and mechanisms that may enhance the establishment and long-term survival of Corynebacterium striatum. in the hospital environment. In this study, we investigated the ability of 22 multidrug-resistant C. striatum clinical isolates to adhere to human epithelial cells and to produce biofilm on polystyrene plates, glass and various tracheostomy tubes. We also tested the virulence of these strains on the nematode Caenorhabditis elegans. They showed good adhesion to epithelial human cells after 180 min of infection. The 22 C. striatum were able to produce biofilms on positively and negatively charged abiotic surfaces at 37 °C. They were also able to infect and to kill Caenorhabditis elegans after 5 days of infection. The virulence condition was associated with the presence of SpaDEF operon encoding pili in all strains. This study provides new insights on virulence mechanisms that may contribute to the persistence of C. striatum in the hospital environment, increasing the probability of causing nosocomial infections.
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Infecções por Corynebacterium , Biofilmes , Corynebacterium/genética , Células Epiteliais , Humanos , VirulênciaRESUMO
Rhodococcus equi causes pyogranulomatous pneumonia in domesticated animals and immunocompromised humans. Dietzia spp. are environmental bacteria that have rarely been associated with human infections. R. equi and Dietzia spp. are closely related actinomycetes. Phenotypic discrimination between R. equi and Dietzia on the basis of their Gram stain morphology and colony appearance is problematic. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is a fast, reliable, and cost-effective method for identification of a wide variety of microorganisms. We have evaluated the performance of Bruker Biotyper versus that of Vitek MS for identification of a collection of 154 isolates identified at the source as R. equi that includes isolates belonging to the genus Dietzia PCR amplification of the choE gene, encoding a cholesterol oxidase, and 16S rRNA sequencing were considered the reference methods for R. equi identification. Biotyper identified 131 (85.1%) of the 154 isolates at the species level, and this figure increased to 152 (98.7%) when the species cutoff was reduced from a score of ≥2.000 to ≥1.750. Vitek MS correctly identified at the species level 130 (84.4%) isolates as long as bacteria were extracted with ethanol but only 35 (22.7%) isolates when samples were prepared by direct extraction from colonies. The two systems allowed differentiation between R. equi and Dietzia spp., but identification of all Dietzia sp. isolates at the species level needed sequencing of the 16S rRNA gene.
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Actinobacteria/classificação , Actinobacteria/isolamento & purificação , Técnicas Bacteriológicas/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Actinobacteria/química , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Humanos , RNA Ribossômico 16S/genética , Sensibilidade e Especificidade , Análise de Sequência de DNARESUMO
The current classifications of "Lisfranc injury" can be purely ligamentous (low-grade midfoot sprains) or involve the osseous and articular structures (high-grade Lisfranc fracture displacements). The first type is often difficult to detect. If these patients are not properly treated, long-term disability can result. The rate of missed or delayed diagnoses has ranged from 13% to 24%, primarily owing to the subtlety of the radiographic findings. This is relatively more common in cases of subtle ligamentous injury (19%). The aim of the present report was to provide a new technique for missed or delayed Lisfranc injury without degenerative local signs. The Lisfranc ligament complex reconstruction is performed with a gracilis tendon graft and is protected by temporary screw fixation. We performed this technique in 3 patients. All 3 patients obtained good results, have been able to resume their previous activities, and have stated they would undergo this type of procedure again. The minimum follow-up length was 2 years.
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Articulações do Pé/lesões , Fixação Interna de Fraturas/métodos , Músculo Grácil/transplante , Ligamentos Articulares/cirurgia , Procedimentos de Cirurgia Plástica/métodos , Retalhos Cirúrgicos/transplante , Adulto , Parafusos Ósseos , Doença Crônica , Diagnóstico Tardio , Erros de Diagnóstico , Feminino , Seguimentos , Articulações do Pé/diagnóstico por imagem , Articulações do Pé/cirurgia , Fixação Interna de Fraturas/instrumentação , Músculo Grácil/irrigação sanguínea , Humanos , Ligamentos Articulares/diagnóstico por imagem , Ligamentos Articulares/lesões , Estudos de Amostragem , Retalhos Cirúrgicos/irrigação sanguínea , Resultado do Tratamento , Adulto JovemRESUMO
Gene sequencing in back of reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is the current approach for discriminating infections produced by different severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants in the clinic. However, sequencing is often a time-consuming step, which hinders the deployment of a very fast response during a pandemic. Here, we propose to run a CRISPR-Cas12a reaction after completing the RT-qPCR and in the very same pot to detect with high specificity genetic marks characterizing variants of concern. A crRNA was appropriately designed to detect the S gene of the SARS-CoV-2 Omicron BA.1 variant. A significant response with >20-fold dynamic range was obtained for the Omicron BA.1 S gene, while the Delta S gene did not produce any detectable signal. The sensitivity of the method was analyzed with a series of diluted samples and different Cas12a nucleases. A correlation between the RT-qPCR CT values and the CRISPR-Cas12a reaction signals was observed. Variant discrimination with the CRISPR-Cas12a reaction was possible in some minutes with high accuracy from patient samples. In conclusion, CRISPR-Cas systems seem ready to be exploited in the clinic to boost personalized diagnoses and accelerate epidemiological surveillance in a cost-effective way.
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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.
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We report the genome of the facultative intracellular parasite Rhodococcus equi, the only animal pathogen within the biotechnologically important actinobacterial genus Rhodococcus. The 5.0-Mb R. equi 103S genome is significantly smaller than those of environmental rhodococci. This is due to genome expansion in nonpathogenic species, via a linear gain of paralogous genes and an accelerated genetic flux, rather than reductive evolution in R. equi. The 103S genome lacks the extensive catabolic and secondary metabolic complement of environmental rhodococci, and it displays unique adaptations for host colonization and competition in the short-chain fatty acid-rich intestine and manure of herbivores--two main R. equi reservoirs. Except for a few horizontally acquired (HGT) pathogenicity loci, including a cytoadhesive pilus determinant (rpl) and the virulence plasmid vap pathogenicity island (PAI) required for intramacrophage survival, most of the potential virulence-associated genes identified in R. equi are conserved in environmental rhodococci or have homologs in nonpathogenic Actinobacteria. This suggests a mechanism of virulence evolution based on the cooption of existing core actinobacterial traits, triggered by key host niche-adaptive HGT events. We tested this hypothesis by investigating R. equi virulence plasmid-chromosome crosstalk, by global transcription profiling and expression network analysis. Two chromosomal genes conserved in environmental rhodococci, encoding putative chorismate mutase and anthranilate synthase enzymes involved in aromatic amino acid biosynthesis, were strongly coregulated with vap PAI virulence genes and required for optimal proliferation in macrophages. The regulatory integration of chromosomal metabolic genes under the control of the HGT-acquired plasmid PAI is thus an important element in the cooptive virulence of R. equi.
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Evolução Molecular , Genes Bacterianos/genética , Rhodococcus equi/patogenicidade , Adaptação Fisiológica/genética , Animais , Cromossomos Bacterianos/genética , Duplicação Gênica/genética , Redes Reguladoras de Genes/genética , Transferência Genética Horizontal/genética , Loci Gênicos/genética , Genômica , Espaço Intracelular/microbiologia , Cinética , Macrófagos/citologia , Macrófagos/microbiologia , Camundongos , Mutação/genética , Filogenia , Plasmídeos/genética , Rhodococcus equi/genética , Rhodococcus equi/crescimento & desenvolvimento , Rhodococcus equi/ultraestrutura , Virulência/genéticaRESUMO
The identification of 83 Corynebacterium, 13 Arcanobacterium haemolyticum, and 10 Rhodococcus equi strains by conventional methods (API Coryne complemented with 16S rRNA gene sequence analysis) was compared with matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry identification. The correlation between API and MALDI-TOF results was 89%. MALDI-TOF is a rapid and accurate system for identification of the above-mentioned microorganisms.
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Arcanobacterium/isolamento & purificação , Técnicas Bacteriológicas/métodos , Corynebacterium/isolamento & purificação , Rhodococcus equi/isolamento & purificação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Animais , Arcanobacterium/química , Arcanobacterium/classificação , Corynebacterium/química , Corynebacterium/classificação , Humanos , Rhodococcus equi/química , Rhodococcus equi/classificação , Sensibilidade e Especificidade , Fatores de TempoRESUMO
Corynebacterium amycolatum is a nonlipophilic coryneform which is increasingly being recognized as a relevant human and animal pathogen showing multidrug resistance to commonly used antibiotics. However, little is known about the molecular mechanisms involved in transition from colonization to the MDR invasive phenotype in clinical isolates. In this study, we performed a comprehensive pan-genomic analysis of C. amycolatum, including 26 isolates from different countries. We obtained the novel genome sequences of 8 of them, which are multidrug resistant clinical isolates from Spain and Tunisia. They were analyzed together with other 18 complete or draft C. amycolatum genomes retrieved from GenBank. The species C. amycolatum presented an open pan-genome (α = 0.854905), with 3,280 gene families, being 1,690 (51.52%) in the core genome, 1,121 related to accessory genes (34.17%), and 469 related to unique genes (14.29%). Although some classic corynebacterial virulence factors are absent in the species C. amycolatum, we did identify genes associated with immune evasion, toxin, and antiphagocytosis among the predicted putative virulence factors. Additionally, we found genomic evidence for extensive acquisition of antimicrobial resistance genes through genomic islands.
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The vegetal world constitutes the main factory of chemical products, in particular secondary metabolites like phenols, phenolic acids, terpenoids, and alkaloids. Many of these compounds are small molecules with antibacterial activity, although very few are actually in the market as antibiotics for clinical practice or as food preservers. The path from the detection of antibacterial activity in a plant extract to the practical application of the active(s) compound(s) is long, and goes through their identification, purification, in vitro and in vivo analysis of their biological and pharmacological properties, and validation in clinical trials. This review presents an update of the main contributions published on the subject, focusing on the compounds that showed activity against multidrug-resistant relevant bacterial human pathogens, paying attention to their mechanisms of action and synergism with classical antibiotics.
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Acinetobacter baumannii is a Gram-negative coccoid rod species, clinically relevant as a human pathogen, included in the ESKAPE group. Carbapenem-resistant A. baumannii (CRAB) are considered by the World Health Organization (WHO) as a critical priority pathogen for the research and development of new antibiotics. Some of the most relevant features of this pathogen are its intrinsic multidrug resistance and its ability to acquire rapid and effective new resistant determinants against last-resort clinical antibiotics, mostly from other ESKAPE species. The presence of plasmids and mobile genetic elements in their genomes contributes to the acquisition of new antimicrobial resistance determinants. However, although A. baumannii has arisen as an important human pathogen, information about these elements is still not well understood. Current genomic analysis availability has increased our ability to understand the microevolution of bacterial pathogens, including point mutations, genetic dissemination, genomic stability, and pan- and core-genome compositions. In this work, we deeply studied the genomes of four clinical strains from our hospital, and the reference strain ATCC®19606TM, which have shown a remarkable ability to survive and maintain their effective capacity when subjected to long-term stress conditions. With that, our aim was presenting a detailed analysis of their genomes, including antibiotic resistance determinants and plasmid composition.
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Rhodococcus equi and Dietzia spp. are closely related actinomycetes that show similar phenotypic properties. In humans, R. equi is an opportunistic pathogen associated with severe immunodeficiency. Dietzia spp. are environmental bacteria that have been isolated recently from clinical material and are presumptively associated with human infections. During the last 5 years, 15 bacterial isolates from human clinical samples collected at the Hospital Marqués de Valdecilla, Santander, Spain, were identified as R. equi by the API Coryne test. 16S rRNA gene sequencing confirmed seven isolates to be true R. equi strains, whereas the other eight were identified as members of the genus Dietzia, including Dietzia maris (four isolates), Dietzia natronolimnaea (two isolates), and Dietzia timorensis and Dietzia sp. (one isolate each). The eight Dietzia isolates were highly sensitive to 12 antimicrobial compounds.
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Infecções por Actinomycetales/diagnóstico , Actinomycetales/classificação , Actinomycetales/isolamento & purificação , Filogenia , Actinomycetales/genética , Técnicas de Tipagem Bacteriana , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Humanos , Dados de Sequência Molecular , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , EspanhaRESUMO
PURPOSE OF THE STUDY: Outbreaks of multidrug-resistant bacteria are increasingly reported at the clinical setting. The antimicrobial, anti-biofilm, anti-quorum sensing, and anti-oxidant activities of four essential oils extracted from Cinnamomum verum, Origanum majorana, Thymus vulgaris, and Eugenia caryophyllata against Gram-positive and Gram-negative multidrug-resistant bacteria were evaluated in vitro. MATERIALS AND METHODS: This study was conducted on 105 multidrug resistant clinical strains. Inhibition diameter zone, minimum inhibitory concentration, and minimum bactericide concentration of the oils were determined using agar disc diffusion method and microdilution. The ability of the 4 essential oils to inhibit the production of bacterial biofilms was tested on polystyrene plates, as well as their inhibitory effect on the production of violacein by Chromobacterium violaceum CV026. The anti-oxidant activity was evaluated by the 2,2-diphenyl-1-picrylhydrazyl scavenging method. RESULTS: Essential oils of Cinnamomum verum, Thymus vulgaris and Eugenia caryophyllata showed an important antibacterial activity. The inhibition diameter zone was higher than 20 mm for 90.24 %, 85.71 % and 60.95 % of strains respectively. These essential oils have a remarkable anti-biofilm and anti-quorum sensing activities against almost all the species studied. Clove extract revealed the highest anti-oxidant activity (Pourcentage of inhibtion of DPPH = 90.3 %). CONCLUSION: These results supported the use of the 4 essential oils as alternative or complementary agents to treat infections caused by multidrug-resistant bacteria, and to prevent biofilm formation and quorum sensing signaling. They might be used as a safe anti-oxidants instead of harmful artificial ones.
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Antibacterianos/farmacologia , Antioxidantes/farmacologia , Biofilmes/efeitos dos fármacos , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Óleos Voláteis/farmacologia , Percepção de Quorum/efeitos dos fármacos , Antibacterianos/isolamento & purificação , Antioxidantes/isolamento & purificação , Compostos de Bifenilo , Cinnamomum zeylanicum/química , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Avaliação Pré-Clínica de Medicamentos , Farmacorresistência Bacteriana Múltipla , Indóis/metabolismo , Testes de Sensibilidade Microbiana , Óleos Voláteis/isolamento & purificação , Origanum/química , Picratos , Syzygium/química , Thymus (Planta)/químicaRESUMO
Corynebacterium urealyticum is a non-diphtherial urease-producing clinically relevant corynebacterial, most frequently involved in urinary tract infections. Most of the C. urealyticum clinical isolates are frequently resistant to several antibiotics. We investigated the susceptibility of 40 C. urealyticum isolated in our institution during the period 2005-2017 to eight compounds representative of the main clinically relevant classes of antimicrobial agents. Antimicrobial susceptibility was determined by the Epsilometer test. Resistance genes were searched by PCR. All strains were susceptible to vancomycin whereas linezolid and rifampicin also showed good activity (MICs90 = 1 and 0.4 mg/L, respectively). Almost all isolates (39/40, 97.5%) were multidrug resistant. The highest resistance rate was observed for ampicillin (100%), followed by erythromycin (95%) and levofloxacin (95%). Ampicillin resistance was associated with the presence of the blaA gene, encoding a class A ß-lactamase. The two rifampicin-resistant strains showed point mutations driving amino acid replacements in conserved residues of RNA polymerase subunit ß (RpoB). Tetracycline resistance was due to an efflux-mediated mechanism. Thirty-nine PFGE patterns were identified among the 40 C. urealyticum, indicating that they were not clonally related, but producing sporadic infections. These findings raise the need of maintaining surveillance strategies among this multidrug resistant pathogen.
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An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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OBJECTIVES: Corynebacterium urealyticum is a non-diphtherial urease-producing clinically relevant corynebacterium associated with urinary tract infections. Most clinical C. urealyticum isolates are multidrug-resistant. Whole-genome sequencing (WGS) of C. urealyticum VH4248 isolated from a clinical urine sample at Hospital Universitario Marqués de Valdecilla, Santander, Spain, was performed to predict its antimicrobial resistance profile and to compare it with results of culture-based phenotypic antimicrobial susceptibility testing. METHODS: Classical microbiological methods and VITEK® MS were used for isolation and initial identification of strain VH4248. Draft genome sequencing was performed on an Illumina HiSeq 2500 platform, followed by assembly and annotation using SPAdes and RAST. Resistance genes were identified through PATRIC, the Pathosystems Resource Integration Center. Average nucleotide identity (ANI) analysis was done using the EDGAR and OrthoANI databases. Antimicrobial susceptibility was determined by Etest. RESULTS: Isolate VH4248 was initially identified asC. urealyticum. Its genome size is 2 261 231 bp with 64.4% GC content. Genome-based identification tools showed an average 93.7% similarity between VH4248 and C. urealyticum genomes deposited in public databases. Therefore, this isolate must be classified as Corynebacterium sp. The blaA and ermX genes as well as a class 1 integron including the aadB and sul1 genes are present in the VH4248 genome. This isolate is highly resistant to ampicillin, erythromycin and trimethoprim/sulfamethoxazole, and moderately resistant to gentamicin and kanamycin. CONCLUSIONS: WGS is a powerful tool forCorynebacterium identification to species level and for detection of unusual resistance determinants, such as that encoded by the class 1 integron in isolate VH4248.
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Antibacterianos , Corynebacterium , Antibacterianos/farmacologia , Corynebacterium/genética , Testes de Sensibilidade Microbiana , EspanhaRESUMO
Exopolysaccharides play an important role in the rheology and texture of fermented foods, and among these beta-glucans have immunomodulating properties. We show that the overproduction of the Pediococcus parvulus GTF glycosyltransferase in an uncapsulated Lactococcus lactis strain results in synthesis and secretion (300 mg liter(-1)) of a position 2-substituted (1-->3)-beta-D-glucan that has potential use as a food additive.