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PURPOSE: Campylobacter is a frequent cause of enteric infections with common antimicrobial resistance issues. The most recent reports of campylobacteriosis in Italy include data from 2013 to 2016. We aimed to provide national epidemiological and microbiological data on human Campylobacter infections in Italy during the period 2017-2021. METHODS: Data was collected from 19 Hospitals in 13 Italian Regions. Bacterial identification was performed by mass spectrometry. Antibiograms were determined with Etest or Kirby-Bauer (EUCAST criteria). RESULTS: In total, 5419 isolations of Campylobacter spp. were performed. The most common species were C. jejuni (n = 4535, 83.7%), followed by C. coli (n = 732, 13.5%) and C. fetus (n = 34, 0.6%). The mean age of patients was 34.61 years and 57.1% were males. Outpatients accounted for 54% of the cases detected. Campylobacter were isolated from faeces in 97.3% of cases and in 2.7% from blood. C. fetus was mostly isolated from blood (88.2% of cases). We tested for antimicrobial susceptibility 4627 isolates (85.4%). Resistance to ciprofloxacin and tetracyclines was 75.5% and 54.8%, respectively; resistance to erythromycin was 4.8%; clarithromycin 2% and azithromycin 2%. 50% of C. jejuni and C. coli were resistant to ≥ 2 antibiotics. Over the study period, resistance to ciprofloxacin and tetracyclines significantly decreased (p < 0.005), while resistance to macrolides remained stable. CONCLUSION: Campylobacter resistance to fluoroquinolones and tetracyclines in Italy is decreasing but is still high, while macrolides retain good activity.
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Antibacterianos , Infecções por Campylobacter , Campylobacter , Testes de Sensibilidade Microbiana , Humanos , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/microbiologia , Itália/epidemiologia , Feminino , Masculino , Adulto , Antibacterianos/farmacologia , Pessoa de Meia-Idade , Adulto Jovem , Adolescente , Idoso , Campylobacter/efeitos dos fármacos , Campylobacter/isolamento & purificação , Criança , Pré-Escolar , Lactente , Fezes/microbiologia , Farmacorresistência Bacteriana , Idoso de 80 Anos ou mais , Recém-Nascido , Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/isolamento & purificaçãoRESUMO
BACKGROUND: Since February 2021 active screening of COVID-19-associated pulmonary aspergillosis (CAPA) has been implemented in our institution. OBJECTIVES: To evaluate CAPA incidence in our centre and evaluate performance of our screening protocol. METHODS: We screened once per week, collecting endotracheal aspirates for fungal culture and galactomannan (GM) and serum for 1,3-ß-D-glucan (BG). In case of positivity (GM more than 4.5, platelia assay, and/or BG >7 pg/ml, wako and/or positive fungal culture), second-level investigations were performed to pursue CAPA diagnosis according to ECMM/ISHAM criteria: bronchoalveolar lavage (BAL) fungal culture and GM, chest computed tomography (CT), serum GM. RESULTS: A total of 102 patients were screened (median age 64 years, range 39-79; 28 (27.4%) females). Twenty-two patients were diagnosed with CAPA (21%). 12 patients were positive for serum BG, 17 patients were positive for endotracheal aspirates GM and 27 patients were positive for endotracheal aspirates fungal culture. Thirty-two BALs were performed, and 26 patients underwent CT chest. Following the second level investigations 61% of the patients with positive screening tests were diagnosed with CAPA. Serum BG above 20 pg/ml or positive serum GM were always associated with typical CT chest signs of aspergillosis. Compared with 1 single positive test, having 2 positive screening test was significantly more associated with CAPA diagnosis (p = .0004). CONCLUSIONS: Active CAPA screening with serum 1,3-ß-D-glucan and endotracheal aspirates galactomannan and fungal cultures and consequent second level investigations led to high number of CAPA diagnosis. Combining more positive fungal biomarkers was more predictive of CAPA diagnosis.
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COVID-19 , Aspergilose Pulmonar Invasiva , Aspergilose Pulmonar , beta-Glucanas , Feminino , Humanos , Adulto , Pessoa de Meia-Idade , Idoso , Masculino , Aspergilose Pulmonar Invasiva/diagnóstico , Aspergilose Pulmonar Invasiva/complicações , COVID-19/complicações , Aspergilose Pulmonar/diagnóstico , Aspergilose Pulmonar/complicações , Mananas , Líquido da Lavagem Broncoalveolar/microbiologia , Sensibilidade e EspecificidadeRESUMO
Introduction: Fusobacterium nucleatum is a gram-negative anaerobe, a constituent of the oral microflora, responsible for chronic periodontal diseases. Case Report: We describe a preterm infant with premature rupture of membranes at 23 weeks of gestational age due to F. nucleatum. The newborn died soon after birth. Placental histopathology showed severe necrotizing chorioamnionitis and funisitis with gram-negative bacilli. After autopsy, F. nucleatum was microbiologically isolated from the lung. The mother had dental hygiene 1 day before delivery, presenting mild and diffuse gingivitis. At admission, she had leukocytosis, foul-smelling vaginal discharge, but no fever. Conclusion: This case highlights the possibility of F. nucleatum spreading from oral cavity after a dental procedure to the placenta with chorioamnionitis and fetal infection. This raises the question of whether dental procedures during pregnancy should be accompanied by prophylactic antibiotics.
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Corioamnionite , Morte Perinatal , Sepse , Gravidez , Humanos , Recém-Nascido , Feminino , Placenta/patologia , Corioamnionite/microbiologia , Fusobacterium nucleatum , Recém-Nascido PrematuroRESUMO
Freezing and thawing have the potential to alter the gross and histologic appearance of tissues, causing damage to individual cells and disrupting the overall architecture. In forensic investigations, freezing and thawing can play a crucial role in cases of unknown cause of death. Perpetrators may use freezing preservation to conceal the body or obscure the time of death. Freezing can also occur naturally when a body is exposed to the elements, sometimes even leading to death itself. We present a case report involving an autopsy performed on an infant, who died of natural causes, after undergoing freezing and thawing. The objective of this study was to identify and discuss the histological artifacts observed in different tissues as a result of the freeze-thaw process. Histologically, the infant's tissues exhibited the most common features described in the literature. Ice crystal artifacts, characterized by expansion of the extracellular space and tissue clefts, were found in the heart, brain, liver, lungs, and kidneys. On the contrary, adipose tissue was not affected, likely due to the scarcity of water. Freeze-thaw artifacts should be taken into account whether a body is known to have been frozen or to add further data if found already defrosted.
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INTRODUCTION: A lack of updated data on the burden and profile of anaerobic bloodstream infections (ABIs) exists. We assessed the incidence of ABIs and trends in antimicrobial resistance in anaerobes isolated from blood in Italy. MATERIAL AND METHODS: We conducted a retrospective study on 17 Italian hospitals (2016-2020). Anaerobes isolated from blood culture and their in vitro susceptibility profiles (EUCAST-interpreted) were registered and analyzed. RESULTS: A total of 1960 ABIs were identified. The mean age of ABIs patients was 68.6 ± 18.5 years, 57.6% were males. The overall incidence rate of ABIs was 1.01 per 10.000 patient-days. Forty-seven% of ABIs occurred in medical wards, 17% in ICUs, 14% in surgical wards, 7% in hemato-oncology, 14% in outpatients. The three most common anti-anaerobic tested drugs were metronidazole (92%), clindamycin (89%) and amoxicillin/clavulanate (83%). The three most common isolated anaerobes were Bacteroides fragilis (n = 529), Cutibacterium acnes (n = 262) and Clostridium perfringens (n = 134). The lowest resistance rate (1.5%) was to carbapenems, whereas the highest rate (51%) was to penicillin. Clindamycin resistance was >20% for Bacteroides spp., Prevotella spp. and Clostridium spp. Metronidazole resistance was 9.2% after excluding C. acnes and Actinomyces spp. Bacteroides spp. showed an increased prevalence of clindamycin resistance through the study period: 19% in 2016, 33% in 2020 (p ≤ 0.001). CONCLUSIONS: Our data provide a comprehensive overview of the epidemiology of ABIs in Italy, filling a gap that has existed since 1995. Caution is needed when clindamycin is used as empirical anti-anaerobic drug.
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Infecções Bacterianas , Sepse , Idoso , Idoso de 80 Anos ou mais , Anaerobiose , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Bactérias Anaeróbias , Infecções Bacterianas/microbiologia , Clindamicina , Farmacorresistência Bacteriana , Feminino , Humanos , Masculino , Metronidazol , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
This study aims to describe trends of mcr-positive Enterobacterales in humans based on laboratory surveillance with a defined catchment population. The data source is the Micro-RER surveillance system, established in Emilia-Romagna region (Italy), to monitor the trend of mcr resistance. Enterobacterales isolates from human clinical samples with minimum inhibitory concentration (MIC) ≥ 2 mg/L for colistin were sent to the study reference laboratory for the detection of mcr genes. Isolates prospectively collected in the period 2018-2020 were considered for the assessment of population rates and trends; further analyses were carried out for the evaluation of clonality and horizontal mcr gene transfer. Previous isolates from local laboratory collection were also described. In the period 2018-2020, 1164 isolates were sent to the reference laboratory, and 51 (4.4%) were confirmed as mcr-positive: 50 mcr-1 (42 Escherichia coli, 6 Klebsiella pneumoniae, 2 Salmonella enterica) and 1 mcr-4 (Enterobacter cloacae). The number of mcr-positive isolates dropped from 24 in the first half of 2018 to 3 in the whole of 2020 (trend p value < 0.001). Genomic analyses showed the predominant role of the horizontal transfer of mcr genes through plasmids or dissemination of transposable elements compared to clonal dissemination of mcr-positive microorganisms. The study results demonstrate a substantial decrease in the circulation of mcr-1 plasmid genes in Emilia-Romagna Region.
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Proteínas de Bactérias/metabolismo , Infecções por Enterobacteriaceae/microbiologia , Enterobacteriaceae/enzimologia , Etanolaminofosfotransferase/metabolismo , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Farmacorresistência Bacteriana , Enterobacteriaceae/classificação , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/genética , Infecções por Enterobacteriaceae/epidemiologia , Etanolaminofosfotransferase/genética , Humanos , Itália/epidemiologia , Testes de Sensibilidade Microbiana , Filogenia , Estudos RetrospectivosRESUMO
This multicenter study evaluated the performance of the Cepheid Xpert Carba-R assay, a qualitative PCR test designed for the rapid detection of blaKPC, blaNDM, blaVIM, blaIMP, and blaOXA-48 carbapenem resistance genes from bacterial isolates grown on blood agar or MacConkey agar. The results were compared to those obtained from bidirectional DNA sequence analysis of nucleic acid extracted from pure colonies. Isolates of Enterobacteriaceae, Pseudomonas aeruginosa, and Acinetobacter baumannii that tested as either intermediate or resistant to a carbapenem antibiotic were analyzed. A total of 467 isolates were evaluated, including prospectively collected clinical isolates, frozen isolates, and a group of contrived broth specimens sent by a central reference laboratory. The assay was run on the GeneXpert platform and took 48 min, with less than 1 min of hands-on time. Compared to the results of the reference methods, the overall sensitivity of the assay was 100% (95% confidence interval [CI], 99.0 to 100%) for isolates grown on both blood and MacConkey agars. Overall specificity was 98.1% (95% CI, 93.1 to 99.8%) and 97.1% (95% CI, 91.7 to 99.4%) for blood and MacConkey agars, respectively. This platform, previously demonstrated to be effective for the detection of carbapenemase genes in rectal swabs, is also adequate for the detection of these genes in bacterial colonies isolated from blood and MacConkey agars.
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Proteínas de Bactérias/genética , Bactérias Gram-Negativas/isolamento & purificação , Infecções por Bactérias Gram-Negativas/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , beta-Lactamases/genética , Antibacterianos/farmacologia , Carbapenêmicos/farmacologia , Farmacorresistência Bacteriana/genética , Bactérias Gram-Negativas/enzimologia , Bactérias Gram-Negativas/genética , Humanos , Illinois , Itália , Testes de Sensibilidade Microbiana , Técnicas de Diagnóstico Molecular/normas , Oregon , Reação em Cadeia da Polimerase em Tempo Real/normas , Sensibilidade e Especificidade , Espanha , Fatores de TempoRESUMO
The global spread of multidrug-resistant Gram-negative bacteria has led to the return of colistin for treating severe infections. Recently, different plasmid-mediated genes conferring resistance to this drug were described and reported worldwide. International committees (EUCAST/CLSI) reevaluated inconsistencies surrounding colistin antimicrobial susceptibility testing (AST), concluding that broth microdilution (BMD) should serve as the reference method for AST. The development of an accurate, reproducible commercial test based on BMD is therefore highly desirable. SensiTest Colistin (STC), a BMD-based compact 4-test panel containing the lyophilized antibiotic in 7 2-fold dilutions (0.25 to 16 µg/ml) was here compared with the EUCAST-CLSI standard reference method (BMD) and, for some isolates, with the automated Phoenix 100 system (PHX). A total of 353 bacterial strains were evaluated by two different laboratories; 137 isolates were resistant to colistin (19 were intrinsically resistant, 83 harbored the mcr-1 gene). Essential agreement (EA) between STC and BMD was obtained for 339 out of the 353 strains tested (96.0%). Overall categorical agreement was obtained for 349 out of the 353 strains analyzed (98.9%). Two major errors (MEs; 0.93%) and two very major errors (VMEs; 1.46%) were documented. STC appeared to be a simple but highly reliable test with good reproducibility even with panels stored at room temperature or at 35°C. Moreover, STC showed a good performance with strains carrying the mcr-1 gene, with a 98.8% EA. As the secondary endpoint of our study, VMEs for PHX were documented for 6 isolates (10%).
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Antibacterianos/farmacologia , Colistina/farmacologia , Bactérias Gram-Negativas/efeitos dos fármacos , Testes de Sensibilidade Microbiana/métodos , Acinetobacter baumannii , Carbapenêmicos/farmacologia , Farmacorresistência Bacteriana Múltipla , Humanos , Testes de Sensibilidade Microbiana/normas , Plasmídeos/genética , Reprodutibilidade dos TestesRESUMO
Objectives: In this study we compared the recently described mcr-4-positive Salmonella enterica monophasic variant, isolated in 2016 in two Italian patients affected by gastroenteritis, with the first mcr-4-positive Salmonella isolate identified in 2013 in a pig at slaughter in Italy. Methods: WGS of the two Salmonella isolates of human origin was performed using a MiSeq instrument (Illumina). The phylogenetic analysis was performed by SNP analysis, comparing genomes of the mcr-4-positive isolates of swine and human origin with 82 Salmonella genomes downloaded from the EnteroBase Salmonella database. Complete sequences of plasmids carrying mcr-4.2 were obtained and compared. Transformation experiments were performed to transfer the mcr-4 plasmids into a colistin-susceptible Escherichia coli recipient strain. Results: Comparative genomics demonstrated that the Salmonella of swine origin did not cluster with the isolates of human origin. The mcr-4.2 gene variant identified in the Salmonella of human origin was located on a ColE-like plasmid. This plasmid showed different replication and mobilization genes with respect to those previously described in the ColE plasmid carrying the mcr-4.1 variant, identified in Salmonella of swine origin. Conclusions: The divergence in genomes, plasmids and gene variants demonstrated that there was not a unique mcr-4-positive, monophasic Salmonella lineage circulating in animals and causing gastroenteritis in humans in Italy. There was no horizontal transfer of the same plasmid among Salmonella strains of animal and human origin, but the mcr-4 gene and a fragment of the plasmid identified in the animal strain were mobilized by an IS1294 into a different ColE plasmid.
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Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Colistina/farmacologia , Variação Genética , Salmonella enterica/efeitos dos fármacos , Salmonella enterica/genética , Animais , Hibridização Genômica Comparativa , Farmacorresistência Bacteriana , Escherichia coli/genética , Gastroenterite/microbiologia , Genes Bacterianos , Humanos , Itália , Testes de Sensibilidade Microbiana , Filogenia , Plasmídeos , Polimorfismo de Nucleotídeo Único , Suínos/microbiologia , Sequenciamento Completo do GenomaRESUMO
In this study we report the detection of the recently described mcr-4 gene in two human isolates of Salmonella enterica serovar Typhimurium. The strains were isolated from faecal samples of two Italian patients with gastroenteritis, collected in 2016. The identified mcr-4 genes (variant mcr-4.2) differed from the mcr-4 gene originally described in a Salmonella strain of swine origin from Italy. Salmonella species could represent a hidden reservoir for mcr genes.
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Antibacterianos/farmacologia , Farmacorresistência Bacteriana/genética , Fezes/microbiologia , Gastroenterite/diagnóstico , Salmonella typhimurium/genética , Salmonella typhimurium/isolamento & purificação , Proteínas de Bactérias/genética , Colistina/farmacologia , Gastroenterite/microbiologia , Genes Bacterianos , Humanos , Itália , Testes de Sensibilidade Microbiana , Salmonella typhimurium/efeitos dos fármacos , Análise de Sequência de DNA , SorogrupoRESUMO
Carbapenemase-producing organisms (CPO) have been identified by global health leaders as an urgent threat. Detection of patients with gastrointestinal carriage of CPO is necessary to interrupt their spread within health care facilities. In this multisite study, we assessed the performance of the Xpert Carba-R test, a rapid real-time quantitative PCR (qPCR) assay that detects five families of carbapenemase genes (blaIMP, blaKPC, blaNDM, blaOXA-48, and blaVIM) directly from rectal swab specimens. Using dual swabs, specimens from 755 patients were collected and tested prospectively. An additional 432 contrived specimens were prepared by seeding well-characterized carbapenem-susceptible and -nonsusceptible strains into a rectal swab matrix and inoculating them onto swabs prior to testing. Antimicrobial susceptibility testing, broth enriched culture, and DNA sequencing were performed by a central laboratory blind to the Xpert Carba-R results. The Xpert Carba-R assay demonstrated a positive percentage of agreement (PPA) between 60 and 100% for four targets (blaKPC, blaNDM, blaVIM, and blaOXA-48) and a negative percentage of agreement (NPA) ranging between 98.9 and 99.9% relative to the reference method (culture and sequencing of any carbapenem-nonsusceptible isolate). There were no prospective blaIMP-positive samples. Contrived specimens demonstrated a PPA between 95 and 100% and an NPA of 100% for all targets. Testing of rectal swabs directly using the Xpert Carba-R assay is effective for rapid detection and identification of CPO from hospitalized patients.
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Técnicas Bacteriológicas/métodos , Portador Sadio/diagnóstico , Bactérias Gram-Negativas/genética , Infecções por Bactérias Gram-Negativas/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Resistência beta-Lactâmica , Portador Sadio/microbiologia , Bactérias Gram-Negativas/isolamento & purificação , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Valor Preditivo dos Testes , Estudos Prospectivos , Reto/microbiologia , Fatores de TempoRESUMO
BACKGROUND: Antimicrobial resistance is a growing threat to public health. Pseudomonas aeruginosa is a relevant pathogen causing human and animal infections, frequently displaying high levels of resistance to commonly used antimicrobials. The increasing difficulty to develop new effective antibiotics have discouraged investment in this area and only a few new antibiotics are currently under development. An approach to overcome antibiotic resistance could be based on antimicrobial peptides since they offer advantages over currently used microbicides. METHODS: The antimicrobial activity of the synthetic peptide AMP2041 was evaluated against 49 P. aeruginosa clinical strains with high levels of antimicrobial resistance, isolated from humans (n = 19) and animals (n = 30). In vitro activity was evaluated by a microdilution assay for lethal dose 90% (LD90), while the activity over time was performed by time-kill assay with 12.5 µg/ml of AMP2014. Evidences for a direct membrane damage were investigated on P. aeruginosa ATCC 27853 reference strain, on animal isolate PA-VET 38 and on human isolate PA-H 24 by propidium iodide and on P. aeruginosa ATCC 27853 by scanning electron microscopy. RESULTS: AMP2041 showed a dose-dependent activity, with a mean (SEM) LD90 of 1.69 and 3.3 µg/ml for animal and human strains, respectively. AMP2041 showed microbicidal activity on P. aeruginosa isolates from a patient with cystic fibrosis (CF) and resistance increased from first infection isolate (LD90 = 0.3 µg/ml) to the mucoid phenotype (LD90 = 10.4 µg/ml). The time-kill assay showed a time-dependent bactericidal effect of AMP2041 and LD90 was reached within 20 min for all the strains. The stain-dead assay showed an increasing of membrane permeabilization and SEM analysis revealed holes, dents and bursts throughout bacterial cell wall after 30 min of incubation with AMP2041. CONCLUSIONS: The obtained results assessed for the first time the good antimicrobial activity of AMP2041 on P. aeruginosa strains of human origin, including those deriving from a CF patient. We confirmed the excellent antimicrobial activity of AMP2041 on P. aeruginosa strains derived from dog otitis. We also assessed that AMP2041 antimicrobial activity is linked to changes of the P. aeruginosa cell wall morphology and to the increasing of membrane permeability.
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Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla , Peptídeos/farmacologia , Infecções por Pseudomonas/microbiologia , Infecções por Pseudomonas/veterinária , Pseudomonas aeruginosa/efeitos dos fármacos , Animais , Membrana Celular/efeitos dos fármacos , Membrana Celular/ultraestrutura , Parede Celular/efeitos dos fármacos , Parede Celular/ultraestrutura , Cães , Humanos , Viabilidade Microbiana/efeitos dos fármacos , Microscopia Eletrônica de Varredura , Pseudomonas aeruginosa/isolamento & purificaçãoRESUMO
Klebsiella pneumoniae is at the forefront of antimicrobial resistance for Gram-negative pathogenic bacteria, as strains resistant to third-generation cephalosporins and carbapenems are widely reported. The worldwide diffusion of these strains is of great concern due to the high morbidity and mortality often associated with K. pneumoniae infections in nosocomial environments. We sequenced the genomes of 89 K. pneumoniae strains isolated in six Italian hospitals. Strains were selected based on antibiotypes, regardless of multilocus sequence type, to obtain a picture of the epidemiology of K. pneumoniae in Italy. Thirty-one strains were carbapenem-resistant K. pneumoniae carbapenemase producers, 29 were resistant to third-generation cephalosporins, and 29 were susceptible to the aforementioned antibiotics. The genomes were compared to all of the sequences available in the databases, obtaining a data set of 319 genomes spanning the known diversity of K. pneumoniae worldwide. Bioinformatic analyses of this global data set allowed us to construct a whole-species phylogeny, to detect patterns of antibiotic resistance distribution, and to date the differentiation between specific clades of interest. Finally, we detected an â¼ 1.3-Mb recombination that characterizes all of the isolates of clonal complex 258, the most widespread carbapenem-resistant group of K. pneumoniae. The evolution of this complex was modeled, dating the newly detected and the previously reported recombination events. The present study contributes to the understanding of K. pneumoniae evolution, providing novel insights into its global genomic characteristics and drawing a dated epidemiological scenario for this pathogen in Italy.
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Antibacterianos/uso terapêutico , Carbapenêmicos/uso terapêutico , Farmacorresistência Bacteriana Múltipla/genética , Infecções por Klebsiella/epidemiologia , Klebsiella pneumoniae/genética , Técnicas de Tipagem Bacteriana , Sequência de Bases , Infecção Hospitalar/microbiologia , DNA Bacteriano/genética , Evolução Molecular , Humanos , Itália/epidemiologia , Infecções por Klebsiella/tratamento farmacológico , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/classificação , Klebsiella pneumoniae/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNA , beta-Lactamases/genéticaRESUMO
BACKGROUND: Staphylococcus pseudintermedius is an opportunistic pathogen recognized as the leading cause of skin, ear, and post-operative bacterial infections in dogs and cats. Zoonotic infections have also recently been reported causing endocarditis, infection of surgical wounds, rhinosinusitis, and catheter-related bacteremia. The aim of the present study is to evaluate, for the first time, the pathogenic potential of S. pseudintermedius isolated from a human infection. To this end, strain DSM 25713, which was recently isolated from a wound of a leukemic patient who underwent a bone marrow transplantation, was investigated for biofilm formation and antibiotic-resistance under conditions relevant for wound infection. RESULTS: The effect of pH (5.5, 7.1, and 8.7) and the presence of serum (diluted at 1:2, 1:10, and 1:100) on biofilm formation was assessed through a crystal violet assay. The presence of serum significantly reduced the ability to form biofilm, regardless of the pH value tested. In vitro activity of eight antibiotics against biofilm formation and mature 48 h-old biofilms was comparatively assessed by crystal violet assay and viable cell count, respectively. Antibiotics at sub-inhibitory concentrations reduced biofilm formation in a dose-dependent manner, although cefoxitin was the most active, causing a significant reduction already at 1/8xMIC. Rifampicin showed the highest activity against preformed biofilms (MBEC90: 2xMIC). None of the antibiotics completely eradicated the preformed biofilms, regardless of tested concentrations. Confocal and electron microscopy analyses of mature biofilm revealed a complex "mushroom-like" architecture consisting of microcolonies embedded in a fibrillar extracellular matrix. CONCLUSIONS: For the first time, our results show that human wound-associated S. pseudintermedius is able to form inherently antibiotic-resistant biofilms, suggestive of its pathogenic potential, and consistent with recent reports of zoonotic infections.
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Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Staphylococcus/efeitos dos fármacos , Staphylococcus/patogenicidade , Infecção da Ferida Cirúrgica/microbiologia , Cefoxitina/farmacologia , Farmacorresistência Bacteriana Múltipla , Humanos , Concentração de Íons de Hidrogênio , Rifampina/farmacologia , Infecções Estafilocócicas/microbiologia , Staphylococcus/isolamento & purificação , Staphylococcus/fisiologiaRESUMO
We first observed the phenomenon of small colony variants (SCVs) in a Staphylococcus pseudintermedius sequence type 71 (ST71) strain, isolated from a non-pet owner. Although we found that small-sized colonies share main features with Staphylococcus aureus SCVs, they nevertheless show a novel, particular, and sticky phenotype, whose expression was extremely stable, even after subcultivation.
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Aderência Bacteriana , Resistência a Meticilina , Staphylococcus/efeitos dos fármacos , Staphylococcus/fisiologia , Feminino , Humanos , Fenótipo , Infecções Estafilocócicas/microbiologia , Staphylococcus/crescimento & desenvolvimento , Staphylococcus/isolamento & purificaçãoRESUMO
A beta-hemolytic Lancefield antigen A-, B-, C-, D-, F-, and G-positive Enterococcus durans strain was cultivated from the rectovaginal swab of a pregnant woman who underwent antenatal screening for Streptococcus agalactiae. The isolate raised concern as to what extent similar strains are misrecognized and lead to false diagnosis of group B streptococci.
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Enterococcus/isolamento & purificação , Infecções por Bactérias Gram-Positivas/diagnóstico , Antígenos de Bactérias/análise , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Diagnóstico Diferencial , Feminino , Infecções por Bactérias Gram-Positivas/microbiologia , Hemólise , Humanos , Dados de Sequência Molecular , Gravidez , Diagnóstico Pré-Natal , RNA Ribossômico 16S/genética , Reto/microbiologia , Análise de Sequência de DNA , Sorotipagem , Streptococcus agalactiae/isolamento & purificação , Vagina/microbiologiaRESUMO
Staphylococcus pseudintermedius is a veterinary pathogen that has seldom been described as an agent of human disease. Features of this probably underreported coagulase-positive Staphylococcus species are depicted here through the description of a graft-versus-host disease-related wound infection caused by a multidrug-resistant strain.
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Transplante de Medula Óssea/efeitos adversos , Portador Sadio/veterinária , Doenças do Cão/microbiologia , Resistência a Meticilina , Infecções Estafilocócicas/tratamento farmacológico , Idoso , Animais , Cães , Doença Enxerto-Hospedeiro/microbiologia , Humanos , Masculino , Dados de Sequência Molecular , Infecções Estafilocócicas/diagnóstico , Staphylococcus/efeitos dos fármacos , Infecção dos Ferimentos/microbiologiaRESUMO
Through a CAMP test, we first observed a Staphylococcus delphini strain (ATCC 49172) to release beta-haemolysin. Production of the latter in this coagulase-positive species of the 'Staphylococcus intermedius Group', in fact, has been labeled to be undetermined, thus far. Of course, a wider number of strains have to be investigated in order to define whether this property is constitutive (like in Staphylococcus (pseud)intermedius), or strain-dependent (like in Staphylococcus aureus), and which clinical impact it has; nevertheless, we can state that S. delphini ATCC 49172 indeed produces this toxin.
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Toxinas Bacterianas/metabolismo , Regulação Bacteriana da Expressão Gênica/fisiologia , Proteínas Hemolisinas/metabolismo , Staphylococcus/classificação , Staphylococcus/metabolismo , Toxinas Bacterianas/genética , Proteínas Hemolisinas/genética , Humanos , Staphylococcus/genéticaRESUMO
Mycobacterium chimaera (MC) is an environmental, slowly growing, non-tuberculous mycobacterium (NTM) belonging to Mycobacterium avium complex (MAC), which recently has been linked to severe cardiovascular infections following open heart and vascular surgery. The majority of the diagnostic laboratory tests used in routine are not able to distinguish MC from M. intracellulare (MI), because of the great genetic similarity existing between these two species. The Genotype Mycobacterium NTM-DR™ represents a valid method to differentiate between these species, but it is expensive, requiring also specialized personnel. Recently, MALDI-TOF MS has been proposed to identify relevant NTM. However, a software implementation is required to distinguish between MC and MI, presenting the two microorganisms' overlapping spectra. The present study evaluates the feasibility of applying a MALDI-TOF logarithmic-based analysis in the routine of a clinical microbiology laboratory, and proposes an easy-to-use template spreadsheet to make the results quickly interpretable. The protocol was previously validated through the identification of 87 strains of MC/MI collected from clinical and environmental samples, and it was identified using the GenoType Mycobacterium NTM-DR™ and/or WGS. The proposed protocol provides accurate identification for the isolates tested; moreover, it is less expensive and more rapid than sequencing methods and can be implemented with minimum effort.
RESUMO
Citrobacter koseri is a facultative anaerobic, motile, non-spore-forming Gram-negative bacillus, which belongs to the family of Enterobacteriaceae. Severe infections due to Citrobacter spp. have been reported in the urinary tract, respiratory airways, intra-abdominal organs, skin and soft tissue, eye, bone, bloodstream, and central nervous system. In newborns, C. koseri is a well-known cause of meningitis, cerebral abscesses, brain adhesions, encephalitis, and pneumocephalus. Infection can be acquired through vertical maternal transmission or horizontal hospital settings; however, in many cases, the source is unknown. Preterm premature rupture of membranes (PPROM), caused by C. koseri, has rarely been described. Herein, we describe a case of PPROM at 16 weeks and 3 days of gestation, leading to anhydramnios. The parents opted for legal termination of the pregnancy, as the prognosis was very poor. C. koseri was isolated postmortem from a placental subamniotic swab and parenchymal sample, as well as fetal blood and lung. To the best of our knowledge, this is the first case of early second-trimester PPROM in which C. koseri infection was demonstrated.