Microbiological characterization of neuropathic diabetic foot infection: a retrospective study at a Portuguese tertiary hospital.

Diabetic foot infection imposes a significant burden and is the major cause of nontraumatic limb amputation. Adequate patient management with effective antibiotic therapy is crucial.This retrospective cohort study aimed to characterize the microbiology and resistance patterns of moderate to severe neuropathic diabetic foot infection in patients hospitalized at a tertiary referral hospital between January 2020 and June 2023. Deep tissue specimens from ulcers were collected for culture.Sixty inpatients were included (62% male, mean age 59.1 ± 11.5 years). Osteomyelitis was present in 90% of the patients. Among 102 microorganisms (average of 1.91 ± 1.25 pathogens per patient), 60.8% were gram-positive bacteria, 31.4% were gram-negative, 3.92% were anaerobic bacteria, and 3.92% were fungi. Staphylococcus aureus (19%) and Enterococcus faecium (17%) were the most common. Pseudomonas aeruginosa (8%) and bacteria of the Enterobacterales family (24%) accounted for all the isolated gram-negative bacteria. Sixteen percent of Staphylococcus aureus and 67% of coagulase-negative Staphylococci were resistant to methicillin. Resistance to ampicillin was found in 11% of Enterococci. All Pseudomonas aeruginosa isolates were sensitive to piperacillin-tazobactam, ceftazidime, or cefepime. Among the Enterobacterales, resistance rates were 35% for piperacillin-tazobactam, 38% for ceftazidime, 21% for cefepime, and 13% for carbapenems.Although the prevalence of methicillin-resistant staphylococci was lower than that in other studies, carbapenem resistance among gram-negative bacteria warrants attention. This study highlights the importance of understanding local epidemiology for effective diabetic foot infection management and resistance mitigation.

Isolation and genomic analysis of "Metallumcola ferriviriculae" MK1, a Gram-positive, Fe(III)-reducing bacterium from the Soudan Underground Mine, an iron-rich Martian analog site.

The Soudan Underground Mine State Park, found in the Vermilion Iron Range in northern Minnesota, provides access to a ~ 2.7 billion-year-old banded iron formation. Exploratory boreholes drilled between 1958 and 1962 on the 27th level (713 m underground) of the mine intersect calcium and iron-rich brines that have recently been subject to metagenomic analysis and microbial enrichments. Using concentrated brine samples pumped from a borehole depth of up to 55 m, a novel Gram-positive bacterium was enriched under anaerobic, acetate-oxidizing, and Fe(III) citrate-reducing conditions. The isolated bacterium, designated strain MK1, is non-motile, rod-shaped, spore-forming, anaerobic, and mesophilic, with a growth range between 24°C and 30°C. The complete circular MK1 genome was found to be 3,720,236 bp and encodes 25 putative multiheme cytochromes, including homologs to inner membrane cytochromes in the Gram-negative bacterium Geobacter sulfurreducens and cytoplasmic membrane and periplasmic cytochromes in the Gram-positive bacterium Thermincola potens. However, MK1 does not encode homologs of the peptidoglycan (CwcA) and cell surface-associated (OcwA) multiheme cytochromes proposed to be required by T. potens to perform extracellular electron transfer. The 16S rRNA gene sequence of MK1 indicates that its closest related isolate is Desulfitibacter alkalitolerans strain sk.kt5 (91% sequence identity), which places MK1 in a novel genus within the Desulfitibacteraceae family and Moorellales order. Within the Moorellales order, only Calderihabitans maritimus strain KKC1 has been reported to reduce Fe(III), and only D. alkalitolerans can also grow in temperatures below 40°C. Thus, MK1 represents a novel species within a novel genus, for which we propose the name "Metallumcola ferriviriculae" strain MK1, and provides a unique opportunity to study a cytochrome-rich, mesophilic, Gram-positive, spore-forming Fe(III)-reducing bacterium.IMPORTANCEThe Soudan Underground Mine State Park gives access to understudied regions of the deep terrestrial subsurface that potentially predate the Great Oxidation Event. Studying organisms that have been relatively unperturbed by surface conditions for as long as 2.7 billion years may give us a window into ancient life before oxygen dominated the planet. Additionally, studying microbes from anoxic and iron-rich environments can help us better understand the requirements of life in analogous environments, such as on Mars. The isolation and characterization of "Metallumcola ferriviriculae" strain MK1 give us insights into a novel genus and species that is distinct both from its closest related isolates and from iron reducers characterized to date. "M. ferriviriculae" strain MK1 may also act as a model organism to study how the processes of sporulation and germination are affected by insoluble extracellular acceptors, as well as the impact of spores in the deep terrestrial biosphere.

Analysis of the Diversity and Functional Potential of Bacterial Communities in Tuberculomas.

The dynamics of lung microbiota in tuberculosis remains poorly understood. Sequencing of variable regions of the 16S rRNA gene from surgically excised tuberculosis foci and biopsy specimens of normal lung tissue allowed characterization of the diversity and predictive potential of bacterial communities. Taxonomic diversity indices attested to differences in the structure of microbial communities between "healthy" lungs and tuberculomas. The microbial composition of "healthy" lungs varied in taxonomic diversity and was presented by both gram-positive and gram-negative bacteria with sufficiently similar metabolic potential. The microbiota of the examined tuberculomas consisted of Mycobacterium tuberculosis in 99.9% of cases. A significant part of the metabolic pathways predicted by PICRUSt2 included cholesterol catabolism, sulfate assimilation, and various pathways for the biosynthesis of cell wall components.

Diversity and Bioprospection of Gram-positive Bacteria Derived from a Mayan Sinkhole.

Water-filled sinkholes known locally as cenotes, found on the Yucatán Peninsula, have remarkable biodiversity. The primary objective of this study was to explore the biotechnological potential of Gram-positive cultivable bacteria obtained from sediment samples collected at the coastal cenote Pol-Ac in Yucatán, Mexico. Specifically, the investigation aimed to assess production of hydrolytic enzymes and antimicrobial compounds. 16 S rRNA gene sequencing led to the identification of 49 Gram-positive bacterial isolates belonging to the phyla Bacillota (n = 29) and Actinomycetota (n = 20) divided into the common genera Bacillus and Streptomyces, as well as the genera Virgibacillus, Halobacillus, Metabacillus, Solibacillus, Neobacillus, Rossellomorea, Nocardiopsis and Corynebacterium. With growth at 55ºC, 21 of the 49 strains were classified as moderately thermotolerant. All strains were classified as halotolerant and 24 were dependent on marine water for growth. Screening for six extracellular hydrolytic enzymes revealed gelatinase, amylase, lipase, cellulase, protease and chitinase activities in 93.9%, 67.3%, 63.3%, 59.2%, 59.2% and 38.8%, of isolated strains, respectively. The genes for polyketide synthases type I, were detected in 24 of the strains. Of 18 strains that achieved > 25% inhibition of growth in the bacterial pathogen Staphylococcus aureus ATCC 6538, 4 also inhibited growth in Escherichia coli ATCC 35,218. Isolates Streptomyces sp. NCA_378 and Bacillus sp. NCA_374 demonstrated 50-75% growth inhibition against at least one of the two pathogens tested, along with significant enzymatic activity across all six extracellular enzymes. This is the first comprehensive report on the biotechnological potential of Gram-positive bacteria isolated from sediments in the cenotes of the Yucatán Peninsula.

A decade of neonatal sepsis in Stockholm, Sweden: Gram-positive pathogens were four times as common as Gram-negatives.

PURPOSE: To assess Gram-positive bacterial (GPB) bloodstream infection (BSI) in neonates, covering incidence, morbidity, mortality, antimicrobial resistance patterns and biomarkers in Region Stockholm, Sweden between 2006 and 2016. METHODS: A population-based retrospective epidemiological study including infants with GPB-BSI, admitted to the neonatal units at Karolinska University Hospital (KUH). Data were collected from patient records, the Swedish Neonatal Quality Register, the microbiological laboratory at KUH and the Swedish Public Health Agency. RESULTS: We identified 357 infants with GPB-BSI, representing an incidence of 1.47/1000 live births (LB). Group B streptococcus (GBS) was the most common pathogen causing BSI in full-term infants and early-onset sepsis (EOS) (0.20/1000 LB), while coagulase-negative staphylococci (CoNS) were predominant in infants born very preterm and in late-onset sepsis (LOS) (0.79/1000 LB). There were no fatal GBS BSI cases, but 10.2% developed meningitis. The GPB case fatality rate was 9.5% and the sepsis fatality rate 2.8%. In GPB-BSI, 1/10 did not have an elevated C-reactive protein level. Staphylococcus aureus (S. aureus) BSI increased during the study period, but no methicillin or vancomycin resistant strains were found. The antimicrobial resistance (AMR) rate was highest in CoNS isolates. CONCLUSION: GPB-BSI was four times more common than Gram-negative BSI in neonates but resulted in lower mortality rate. GBS was the most common pathogen in full-term infants and in EOS. CoNS was the most common pathogen in LOS and infants born very preterm, and the AMR rate was high in these isolates. The increasing trend of S. aureus BSI indicates a need of further investigation.

Synthesis, antimicrobial and antioxidant activities of pyridyl substituted thiazolyl triazole derivatives

In this present study, 63 different 5-[4-methyl-2-(pyridin-3/4-yl)thiazole-5-yl]-4-substituted-3-substituted benzylthio-4H-1,2,4-triazole derivatives were synthesized, and evaluated for their in vitro antimicrobial activity against various human pathogenic microorganisms and antioxidant activity. The derivatives were synthesized in a multi-step synthesis procedure including triazole and thiazole ring closure reactions, respectively. The synthesized derivatives (A1-24; B1-39) were screened for their antibacterial, antifungal, and antioxidant activities compared to standard agents. The derivatives possessing 3-pyridyl moiety particularly exhibited relatively high antibacterial activity (MIC= < 3.09-500 µg/mL) against Gram-positive bacteria, and compounds possessing 4-pyridyl moiety showed remarkable antioxidant activity

Evaluation of a mass spectrometry and Vitek 2 combined protocol for rapid identification and susceptibility testing of Enterobacterales directly from positive blood cultures / Evaluación de la combinación de espectrometría de masas y el sistema Vitek 2 para la identificación rápida y el estudio de sensibilidad de Enterobacterales directamente desde frascos de hemocultivo positivos

OBJECTIVE: The aim was to evaluate a rapid method which would combine identification and susceptibility testing directly from positive blood cultures for Gram-negative bacilli of the Enterobacterales. MATERIAL AND METHODS: Gram-negative rods from blood cultures were directly identified by MALDI-TOF. Samples with Enterobacterales were selected for direct antimicrobial susceptibility testing by Vitek 2. The results were compared to those obtained with our laboratory's standard method. RESULTS: MALDI-TOF directly from blood cultures identified correctly 83% of the samples. Enterobacterales (n = 68) were identified at gender and species level in 85% of blood cultures with a score >1.7. In general, MICs were obtained after 7 h. MICs of amoxicillin-clavulanate, amikacin and ciprofloxacin showed in almost 50% of the cases after 5 h. CONCLUSIONS: A simple procedure with low cost and reduced working time makes it possible to integrate both identification and susceptibility testing directly from blood cultures. Thus, this protocol could offer advantages when it comes to selection and cost of treatment and patients' clinical outcomes

OBJETIVO: Evaluar un método rápido de identificación y estudio de sensibilidad directamente desde hemocultivos positivos para bacilos gramnegativos del orden Enterobacterales. MATERIAL Y MÉTODOS: Los hemocultivos con bacilos gramnegativos fueron utilizados para identificación mediante MALDI-TOF, seleccionándose aquellos con Enterobacterales para estudio de sensibilidad con Vitek 2. Los resultados fueron comparados con el método estándar del laboratorio. RESULTADOS: MALDI-TOF identificó correctamente el 83% de las muestras obtenidas directamente de hemocultivos. En caso de Enterobacterales (n = 68), el 85% se identificaron a nivel de género y especie con un score > 1,7. El tiempo necesario para la obtención de CMIs fue de 7 horas, reduciéndose a 5 en amoxicilina-clavulánico, amicacina y ciprofloxacino, casi en el 50% de los casos. CONCLUSIONES: Un protocolo simplificado, con bajo coste y reducido tiempo de trabajo, combinando identificación y estudio de sensibilidad directamente desde hemocultivos, podría proporcionar beneficios en la elección y coste del tratamiento, y mejora clínica del paciente

Diversidad bacteriana en aguas mineromedicinales del balneario "Urauco". Pichincha. Ecuador / Bacterial biodiversity in mineromedicinal waters of spa "Urauco". Pichincha. Ecuador

La presencia de manantiales de aguas mineromedicinales en Ecuador se encuentra en relación con el gran número de volcanes que existen en el país, los cuales se han venido utilizando desde épocas remotas como medicamentos por la población. Sin embargo, se desconoce cuál es la biodiversidad bacteriana presente. Por ello, el objetivo del trabajo fue determinar la biodiversidad bacteriana de las aguas mineromedicinales del balneario "Urauco" ubicado en la Provincia de Pichincha. Ecuador. Se tomaron muestras de agua de un volumen de 1 litro en dos punto del balneario. Los muestreos se realizaron en 2 ocasiones durante un periodo de dos año. Para la toma y transporte de las muestras se siguieron las normas ecuatorianas INEN 2 176-98 y INEN 2 169-98. Para la determinación de la biodiversidad bacteriana se utilizó las técnicas de siembra, selección e identificación de bacterias indicados por Barrow y Feltan (2004) y MacFfadin (2004), complementadas con las pruebas bioquímicas contenidas en las galerías API (BioMérieux) y galería Microgen (Microgen corp.). Los resultados señalan un población de bacterias heterótrofas en las aguas del balneario de 3,1 x 102 UFC/mL, con la prevalencia de las bacterias Gram negativa de la clase Gamma Proteobacterias. Se pudo aislar e identificar 64 colonias de bacterias. Los géneros de bacterias encontrados fueron Acidovorax, Aeromonas, Bacillus, Brevundimonas, Budvicia, Citrobacter, Corynebacterium, Ed- wardsiella, Enterococcus, Ewingella, Flavobacterium, Kurthia, Micrococcus, Proteus, Pseudomonas, Psychrobacter, Ralstonia, Staphylococcus, Vibrio y Yokonella. La biodiversidad bacteriana observada fue alta, aunque escasa en número de individuos

The presence of mineromedicinal water springs in Ecuador is related to the large number of volcanoes that exist in the country, which have been used since ancient times as medicines by the population. However, it is unknown what bacterial biodiversity is present. Therefore, the objective of these work was to determine the bacterial biodi- versity of the mineromedicinal waters of “Urauco” spa located in Pichincha Provinces of Ecuador. Water samples of a volume of 1 liter were taken at two points in each spa studied. Sampling was done 2 times over a period of two year. To take and transport the samples, the Ecuadorian standards INEN 2 176-98 and INEN 2 169-98 were followed. For the determination of bacterial biodiversity, the techniques of planting, selection and identification of bacteria indicated by Barrow and Feltan (2004) and MacFfadin (2004) were used, complemented with the bio- chemical tests contained in the API (BioMérieux) galleries and Microgen (Microgen corp.). The results indicate a population of heterotrophic bacteria in the spa waters of 3.1 x 102 CFU / mL, with the prevalence of Gram negative bacteria of the Gamma Proteobacteria class. It was possible to isolate and identify 64 colonies of bacteria. The genera of bacteria were Acidovorax, Aeromonas, Bacillus, Brevundimonas, Budvicia, Citrobacter, Corynebacterium, Edwardsiella, Enterococcus, Ewingella, Flavobacterium, Kurthia, Micrococcus, Proteus, Pseudomonas, Psychrobacter, Ralstonia, Staphylococcus, Vibrio and Yokonella. The bacterial biodiversity observed was high, although low in number

Infections and antimicrobial resistance in an adult intensive care unit in a Brazilian hospital and the influence of drug resistance on the thirty-day mortality among patients with bloodstream infections

Abstract INTRODUCTION: The present study aimed to determine the incidence of health care-associated infections (HCAIs) and identify the main resistant microorganisms in intensive care unit (ICU) patients in a Brazilian university hospital. METHODS: A retrospective cohort study was conducted in a Brazilian teaching hospital between 2012 and 2014. RESULTS: Overall, 81.2% of the infections were acquired in the ICU. The most common resistant pathogenic phenotypes in all-site and bloodstream infections were oxacillin-resistant coagulase-negative staphylococci and carbapenem-resistant Acinetobacter spp. (89.9% and 87.4%; 80.6% and 70.0%), respectively. CONCLUSIONS: There is an urgent need to focus on HCAIs in ICUs in Brazil.

Characteristics of the salivary microbiota in cheilitis granulomatosa

BACKGROUND: Cheilitis granulomatosa (CG) is a disturbing and persistent idiopathic lip swelling. The cause and treatment has not been wholly elucidated. Some reports infer that CG is mainly associated with dental infection but no firm or reliable microbiological evidence has been provided for a causative organism. This study aimed to evaluate whether microorganisms contribute to the etiology of CG in order to inform appropriate treatment op-tions in clinic. MATERIAL AND METHODS: Unstimulated saliva was collected from 15 CG patients who were diagnosed clinically and pathologically and 15 healthy controls (HC). DNA was extracted from the precipitate of the centrifuged saliva for 16s rRNA high-throughput sequencing using the Miseq PE300 platform. The distribution of the microbiome between the two groups was compared. RESULTS: CG patients had a greater microbial flora that was more diverse than the HC. Prevotella, Alloprevotella, Porphyromonas, Actinomyces, Rothia, Fusobacterium, Haemophilus, and Aggregatibacter had a significantly higher abundance in CG patients. In contrast, Streptococcus and Campylobacter were the most abundant genera in HC with a mean relative abundance of 63% and 2%, respectively. The microbiological network indicated that most of the bacteria that were enriched at greater levels in CG patients were likely to be Prevotella, Actinomyces, and Rothia. These have been shown to co-exist with other bacteria. CONCLUSIONS: The composition and structure of bacterial communities in CG patients were different from HC. Most of the genera observed in CG patients were associated with periodontitis and pulp infection. These findings might be helpful in understanding the etiology of CG. Further study will be needed to confirm these findings and explore the underlying pathological mechanism

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Comparison of two methods skipping cell lysis and protein extraction for identification of bacteria from blood cultures by matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry / Comparación de dos métodos que eluden la lisis celular y la extracción de proteínas para la identificación de bacterias crecidas en hemocultivos mediante espectrometría de masas MALDI-TOF

OBJECTIVE: Matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry (MALDI-TOF MS) is widely used for fast identification of bacteria from blood cultures (BC). We compared the performance of two procedures, one including a pre-enrichment step in brain heart infusion and the other a direct method using vacutainer separator gel tubes (DI), for identification of bacteria from blood cultures by MALDI-TOF MS. MATERIAL AND METHODS: We first prepared a training set of 20 simulated bacteremia specimens, including 10 Gram-negative and 10 Gram-positive species. A total of 145 non-consecutive BCs flagged as positive (68 Gram-negative rods, and 77 Gram-positive cocci) were prospectively analyzed (validation set). RESULTS: A total of 82% and 49% of isolates were correctly identified to the species level by the respective methods. CONCLUSION: The pre-enrichment method outperformed the DI method for identification of virtually all bacterial species included in the panels

OBJETIVO: La espectrometría de masas MALDI-TOF se utiliza comúnmente para la identificación rápida de bacterias crecidas en hemocultivos (HC). Hemos comparado el rendimiento de dos procedimientos, uno que incluye un paso previo del enriquecimiento en caldo corazón-cerebro y el otro un método directo que usa tubos vacutainer con gel separador (DI), para la identificación de bacterias a partir de hemocultivos mediante MALDI-TOF MS. MATERIAL Y MÉTODOS: Analizamos prospectivamente un total de 145 HC no consecutivos (68 con crecimiento de bacterias gramnegativas y 77 de cocos grampositivos). RESULTADOS: Un total de 82% y 49% de los aislamientos fueron identificados correctamente a nivel de especie por los dos métodos, respectivamente. CONCLUSIÓN: El rendimiento del método de pre-enriquecimiento en caldo corazón-cerebro fue mejor que el del método DI para la identificación de la práctica totalidad de las especies bacterianas incluidas en el panel de estudio

Direct matrix-assisted laser desorption ionization time-of-flight mass spectrometry and real-time PCR in a combined protocol for diagnosis of bloodstream infections: a turnaround time approach

ABSTRACT Bloodstream infections (BSIs) are serious infections associated with high rates of morbidity and mortality. Every hour delay in initiation of an effective antibiotic increases mortality due to sepsis by 7%. Turnaround time (TAT) for conventional blood cultures takes 48 h, forcing physicians to streamline therapy by exposing patients to broad-spectrum antimicrobials. Our objective was (1) to evaluate the accuracy and TAT of an optimized workflow combining direct matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and in-house real-time polymerase chain reaction (PCR) for bacterial identification and antimicrobial resistance profiling directly from positive blood bottles for diagnosing bloodstream infections and (2) to verify the effect of reporting results to medical staff. A total of 103 BSI episodes from 91 patients admitted to three hospitals in São Paulo, Brazil were included. TAT from molecular versus conventional methods was measured and compared. Our protocol showed an overall agreement of 93.5% for genus and 78.5% for species identification; 74.2% for methicillin resistance detection, 89.2% for extended-spectrum β-lactamase profiling, 77.8% for metallo-β-lactamase profiling, and 100% for carbapenemase profile and vancomycin-resistance detection when compared with conventional testing. TAT of molecular sample processing according to our protocol was 38 h shorter than conventional methods. Antimicrobial interventions were possible in 27 BSI episodes. Antimicrobial discontinuation was achieved in 12 BSI episodes while escalation of therapy occurred in 15 episodes. Antimicrobial therapy was inadequate in three (12%) BSI episodes diagnosed using results of molecular testing. Our in-house rapid protocol for identifying both bacteria and antimicrobial resistance provided rapid and accurate results, having good agreement with conventional testing results. These results could contribute to faster antimicrobial therapy interventions in BSI episodes.

Perfil de resistencia antibiótica en infecciones oculares en un centro de referencia en Floridablanca, Colombia / Antibiotic resistance profile in eye infections in a reference centre in Floridablanca, Colombia

Antecedentes y objetivo: Ante la creciente resistencia bacteriana a los antibióticos es necesario determinarla de manera local para servir de guía en el manejo clínico. El propósito de este estudio fue caracterizar el patrón de sensibilidad antibiótica en casos de infecciones oculares en una institución oftalmológica de tercer nivel en Floridablanca (Colombia). Materiales y métodos: Estudio observacional de corte transversal en el cual se analizaron los informes de cultivo y antibiograma de muestras tomadas de casos de conjuntivitis, queratitis infecciosas y endoftalmitis entre enero de 2013 y junio del 2016. Resultados: Ochocientas treinta y tres muestras fueron positivas para bacterias. Considerando microorganismos tanto grampositivos como gramnegativos, la gentamicina, la tobramicina y la ciprofloxacina mostraron tasas de resistencia altas (64,4; 40,3% y 29,1%). La moxifloxacina, la vancomicina, el imipenem y la gatifloxacina mostraron bajos porcentajes de resistencia: 2,6; 2,1; 0,6 y 0,4% respectivamente. Al comparar los resultados con estudios previos en nuestra institución se evidenció una disminución de la sensibilidad a las quinolonas de cuarta generación y al imipenem, especialmente dentro de los gramnegativos. Conclusión: Las quinolonas de cuarta generación, el imipenem y la vancomicina siguen teniendo in vitro una baja resistencia de las bacterias causantes de infecciones oculares. Sin embargo, se evidenció una tendencia al incremento de la resistencia de las bacterias gramnegativas. Se deben tomar medidas para intentar controlar este fenómeno, y pensar en posibles alternativas de terapia antimicrobiana ante infecciones causadas por estos microorganismos

Background and objective: In view of the increasing bacterial resistance to antibiotics, it is necessary to determine it locally in order to serve as a guide in clinical management. The purpose of this study was to characterise the pattern of antibiotic sensitivity in cases of eye infections in a third level ophthalmological institution in Floridablanca (Colombia). Materials and methods: An observational cross-sectional study in which an analysis was made of the culture and antibiogram reports of specimens taken from cases of conjunctivitis, infectious keratitis, and endophthalmitis between January 2013 and June 2016. Results: A total of 833 specimens were positive for bacteria. Considering both gram-positive and gram-negative microorganisms gentamicin, tobramycin, and ciprofloxacin showed high resistance rates (64.4%, 40.3%, and 29.1%, respectively). Moxifloxacin, vancomycin, imipenem, and gatifloxacin showed low percentages of resistance: 2.6%, 2.1%, 0.6%, and 0.4%, respectively. When comparing the results with previous studies in our institution, there was a decrease in sensitivity to the fourth-generation quinolones and imipenem, especially within the gram-negative ones. Conclusion: Fourth generation quinolones, imipenem and vancomycin continue to have a low in vitro resistance to bacteria that cause eye infections. However, there was a tendency to an increase in the resistance of gram-negative bacteria. Measures should be taken to try to control this phenomenon, and consider possible antimicrobial therapy alternatives to infections caused by these microorganisms

Microbiological analysis of contact lens cases: impact of the hospital environment / Análise microbiológica de estojos de lentes de contato: impacto do ambiente nosocomial

ABSTRACT Purpose: The aim of this study was to analyze the bacterial and fungal microbiota found in contact lens cases among two groups of patients to correlate the data on the habits of contact lens users and to evaluate whether there is a difference in the culture results between users of ridged and nonridged contact lens cases. Methods: Two groups (35 patients per group) were included, consisting of hospital workers and those who had not visited a hospital in the past 30 days, and a questionnaire regarding epidemiological data and habits related to contact lens and lens case use was administered. In addition, 140 samples collected from the right and left compartments of each lens case by swabbing the bottom of the wells were tested using bacterioscopy as well as fungal and bacterial cultures via computerized identification of the species. Results: No fungal growth was identified in any of the 70 contact lens cases; however, bacteria were found in 39 cases, and there was no statistical difference between the groups. Most bacteria (>85%) were gram-negative bacilli. Contamination inone compartment of the contact lens case elevated the risk of contamination of the other side (>80%). Moreover, contamination was statistically higher in the ridged cases than in nonridged cases (p=0.0149). Conclusion: The types of bacteria contaminating the cases are generally not seen in eye diseases associated with contact lens use, suggesting that other decisive variables are involved in eye infection from a contaminated lens or case. Fungal contamination of contact lens cases appears to be an exception. Ridged cases are commonly used by contact lens wearers and present a potential risk to eye health. In addition, the results of bacterial tests between hospital workers and those who did not visit a hospital were not significantly different.

RESUMO Objetivo: O objetivo deste trabalho foi analisar a microbiota bacteriana e fúngica encontrada em estojos de lentes de contato em dois grupos, correlacionar os dados sobre os hábitos de uso de lentes de contato e avaliar se há diferença na positividade das culturas entre os usuários estojos de lentes de contato com ranhuras e sem ranhuras. Métodos: Dois grupos foram formados, trabalhadores do hospital e pessoas que não visitaram o hospital (35 indivíduos por grupo), e um questionário foi aplicado sobre dados epidemiológicos e hábitos relacionados ao uso de lentes de contato e estojos de lentes. Além disso, 140 amostras, coletadas do compartimento direito e esquerdo de cada estojo de lente, esfregando o fundo dos mesmos, foram testadas por bacterioscopia e por culturas de fungos e bactérias, com identificação computadorizada da espécie. Resultados: Não houve crescimento fúngico em nenhum dos 70 estojos de lentes de contato, porém bactérias foram encontradas em 39; não houve diferença estatística entre os grupos. A maioria das bactérias (>85%) eram bacilos gram-negativos. Quando um compartimento estava contaminado, o risco de contaminação do outro compartimento era elevado (>80%). A contaminação foi estatisticamente maior nos estojos com ranhuras (p=0,0149). Conclusão: A contaminação dos estojos parece ocorrer por bactérias que, em geral, não são encontradas em doenças oculares associadas ao uso de lentes de contato, sugerindo que existem outras variáveis decisivas nas infecções oculares de uma lente ou estojo contaminado. Contaminação de estojos de lentes de contato com fungos parece ser uma exceção. O uso de estojos com ranhuras é uma prática comum e apresenta um risco potencial à saúde ocular. Não foram encontradas diferenças significativas nos resultados dos testes bacterianos entre trabalhadores hospitalares e pessoas que não visitaram o hospital.

Epidemiology of diabetic foot infections in a reference tertiary hospital in India

Abstract Introduction The present study attempts to examine the microbial profile and antibiotic susceptibility of diabetic foot infections in the intensive care unit of a tertiary referral centre for diabetic foot. As part of the study, we also attempted to find the prevalence of blaNDM-like gene among carbapenem-resistant gram negative infections. Methodology A prospective study of 261 patients with diabetic foot infections was performed during the period between January 2014 and June 2014. Results A total of 289 isolates were obtained from 178 tissue samples from 261 patients, 156 (59.7%) males and 105 (40.2%) females, with a mean age of 58 years (-15 years), having diabetic foot infection. No growth was seen in thirty eight (17.6%) tissue samples. Out of the total samples, 44.3% were monomicrobial and 55.7% were polymicrobial. Gram negative pathogens were predominant (58.5%). Seven of the total isolates were fungal; 0.7% showed pure fungal growth and 1.7% were mixed, grown along with some bacteria. The most frequently isolated bacteria were Staphylococcus aureus (26.9%), followed by Pseudomonas aeruginosa (20.9%). Of the 58.5% gram negative pathogens, 16.5% were Enterobacteriaceae resistant to carbapenems. Among these isolates, 4 (25%) were positive for blaNDM-like gene. Among the rest, 18.6% were carbapenem-resistant Pseudomonas, among which 4 (36.3%) were blaNDM. Among the Staphylococci, 23.7% were methicillin-resistant Staphylococcus aureus. Conclusions Our results support the recent view that gram negative organisms, depending on the geographical location, may be predominant in DFIs. There is an increase in multidrug-resistant pathogens, especially carbapenem resistance and this is creeping rapidly. We need to be more judicious while using empiric antibiotics.

Cambio de CLSI a EUCAST en la interpretación de la sensibilidad a antimicrobianos: ¿cómo influye en nuestro medio? / From CLSI to EUCAST guidelines in the interpretation of antimicrobial susceptibility: What is the effect in our setting?

INTRODUCCIÓN: La aplicación de los puntos de corte establecidos por el European Committee on Antimicrobial Susceptibility Testing (EUCAST) en comparación con los del Clinical and Laboratory Standards Institute (CLSI) modifica los criterios de interpretación de la sensibilidad de algunos antimicrobianos y esto conduce a cambios en los informes de sensibilidad antibiótica acumulada. MÉTODOS: Análisis de la influencia de la aplicación del EUCAST en 10.359 aislados clínicos de Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus y Enterococcus spp. RESULTADOS: Al aplicar los puntos de corte del EUCAST, la mayoría de los porcentajes de sensibilidad a antimicrobianos no se alteró o lo hizo de forma muy leve; sin embargo, se observó una disminución de la sensibilidad a los aminoglucósidos en bacilos gramnegativos, especialmente a la amicacina en Pseudomonas aeruginosa (23,2%), aunque solo el 5,7% fueron totalmente resistentes; además, disminuyó notablemente el porcentaje de aislados sensibles a aztreonam. Es de destacar el aumento de cepas de Staphylococcus aureus resistentes a clindamicina (51,5%) y a aminoglucósidos (gentamicina 43,1%). CONCLUSIONES: El cambio de los criterios del CLSI a los de EUCAST en algunos patógenos supone una alteración en los porcentajes de resistencia a algunos antimicrobianos y, por tanto, en la epidemiología local de la resistencia. Estos cambios deben realizarse por un grupo multidisciplinar, que analice la influencia de los nuevos datos en los protocolos de tratamiento empírico de cada centro

INTRODUCTION: Implementation of the breakpoints established in the European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines in comparison with those of the Clinical and Laboratory Standards Institute (CLSI) means that the criteria for interpreting the susceptibility of some antimicrobials have been modified, resulting in changes in the reports of accumulated antibiotic susceptibility. METHODS: The effect of applying EUCAST breakpoints in 10,359 clinical isolates of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus spp. was analysed. RESULTS: By applying EUCAST breakpoints, most antimicrobial susceptibility percentages did not change or changed very slightly. However, a decrease in aminoglycoside susceptibility was observed in Gram-negative bacilli, mainly for amikacin and Pseudomonas aeruginosa (23.2%), although only 5.7% were completely resistant; a notably decrease in the percentage of isolates susceptible to aztreonam was also observed. There was also a marked increase in the number of Staphylococcus aureus strains resistant to clindamycin (51.5%) and aminoglycosides (gentamicin 43.1%). CONCLUSIONS: Switching from CLSI to EUCAST criteria in some pathogens alters the percentages of resistance to several antimicrobials, and therefore the local epidemiology of the resistance. These changes should be implemented by a multidisciplinary group in order to analyse the influence of the new data on the empirical treatment protocols of each centre

Microorganismos aislados de hemocultivos en niños con cáncer y neutropenia febril de alto riesgo en cinco hospitales de Santiago, Chile, período 2012-2015 / Microorganisms isolated from blood cultures in children with cancer and high-risk febrile neutropenia from five hospitals in Santiago, Chile, 2012-2015

Resumen Antecedentes: Los microorganismos aislados de hemocultivos (HC) en pacientes con neutropenia febril (NF) varían en el tiempo, siendo necesaria su vigilancia para orientar una terapia empírica adecuada. Objetivo: Identificar microorganismos aislados de HC y su perfil de resistencia (R) a antimicrobianos en niños con cáncer y NF de alto riesgo. Método: Estudio prospectivo, multicéntrico de episodios de NF de alto riesgo en pacientes bajo 18 años de edad, de cinco hospitales en Santiago de Chile, 2012-2015. Análisis de HC positivos. Resultados: Se analizaron 206 microorganismos en 185 episodios de NF de alto riesgo con HC positivos. Los aislados principales fueron bacilos gramnegativos (BGN) (46,6%) y cocáceas grampositivas (CGP) (45,1%) y los microorganismos más frecuentes Escherichia coli (22,8%), Staphylococcus coagulasa negativa (18,0%) y Klebsiella spp (16,5%). En resistencia (R) a antimicrobianos destaca: E. coli y Klebsiella spp 4,2 y 67,6% R a cefalosporinas de tercera generación (cefotaxima/ceftriaxona) respectivamente, 10,6 y 40,6% R a ciprofloxacina y 2,1 y 26,5% a amikacina, respectivamente. S. coagulasa negativa y S. aureus 86,4% y 22,2% R a oxacilina, Streptococcus grupo viridans 71% R a penicilina. Discusión: Este estudio actualiza la etiología y el perfil de R de microorganismos aislados en HC de niños con cáncer y NF de alto riesgo, herramienta esencial para el adecuado manejo de estos pacientes.

Background: Microorganisms isolated from blood cultures (BC) in patients with febrile neutropenia (NF) vary over time, requiring systematic monitoring to guide appropriate empirical therapy. Aim: To identify microorganisms isolated from BC and their antimicrobial resistance profile in children with cancer and high risk NF. Method: Prospective, multicenter study. The analysis included episodes of high-risk FN with positive BC in children under 18 years of age treated in five hospitals in Santiago, Chile, 2012-2015. Results: A total of 206 microorganisms were analyzed in 185 episodes of high-risk FN. The main isolates were Gram negative bacilli (46.6%) and Gram positive cocci (45.1%) and the most frequent microorganisms were Escherichia coli (22.8%), coagulase negative Staphylococcus (18.0%) and Klebsiella spp. (16.5%). Escherichia coli and Klebsiella spp showed 4.2% and 67.6% resistance to third generation cephalosporins (cefotaxime/ceftriaxone), 10.6% and 40.6% resistance to fluoroquinolones (ciprofloxacin) and 2.1% and 26.5% to amikacin, respectively. Coagulase negative Staphylococcus and Staphylococcus aureus had 86.4% and 22.2% resistance to oxacillin, Streptococcus viridans group had 71% resistance to penicillin. Discussion: This study updates the etiology and resistance profile of microorganisms isolated in BC from children with cancer and high risk FN, an essential tool for the adequate management of these patients.

Prevalencia y perfil de susceptibilidad antimicrobiana en bacterias aisladas de úlceras crónicas infectadas en adultos / Prevalence and susceptibility pattern of bacteria isolated from infected chronic wounds in adult patients

Resumen Introducción: Las úlceras crónicas son un problema de salud pública, agravándose por infecciones bacterianas causadas principalmente por agentes resistentes. Objetivo: Estudiar prevalencia y perfil de susceptibilidad en bacterias aisladas de úlceras crónicas en pacientes adultos. Pacientes y Métodos: Pacientes atendidos en la Fundación Instituto Nacional de Heridas entre mayo y julio de 2014, con úlceras crónicas en extremidades inferiores con signos inflamatorios clínicos. Las muestras fueron cultivadas en aerobiosis y anaerobiosis y para la identificación bacteriana se empleó el sistema de galerías API (Biomerieux). La susceptibilidad in vitro se evaluó según el método de Kirby Bauer. Resultados: Se reclutaron 73 pacientes, entre quienes 46 presentaron úlceras infectadas, diagnosticándose 33 úlceras venosas con predominio de infección polimicrobiana y 10 úlceras de pie diabético con predominio de infección monomicrobiana (p ≤ 0,05). Se aislaron 68 cepas de los 46 pacientes con úlcera infectada. Las enterobacterias predominaron en infección monomicrobiana (p ≤ 0,05) y los demás grupos bacterianos fueron levemente más frecuentes en infección polimicrobiana. La especie prevalente fue Staphylococcus aureus (24%) seguida de Pseudomonas aeruginosa (18%). Cincuenta cepas (77%) presentaron resistencia a uno o más antibacterianos. Destacamos resistencia de S. aureus a ciprofloxacina (50%) y cefoxitina (37,5%) identificándose así resistencia a meticilina en la comunidad (SARM-AC), siendo todas sensibles a cotrimoxazol. Las enterobacterias presentaron resistencia a sensibilidad a amikacina (95,5%), P. aeruginosa evidenció resistencia a ciprofloxacina (33,3%) con alta sensibilidad a gentamicina (91,7%) y amikacina (83,3%), mientras Acinetobacter spp presentó resistencia a ciprofloxacina y ceftazidima en 60%, con 100% de sensibilidad a imipenem. Streptococcus β hemolítico presentó 50% de resistencia a clindamicina y penicilina. Conclusión: Estos datos entregan información epidemiológica de infecciones de úlceras crónicas, representando un apoyo al diagnóstico, tratamiento y manejo de esta patología.

Background: Chronic wounds are considered a public health problem that may be complicated by bacterial infections, mainly caused by resistant strains. Aim: To study the bacteria prevalence and antimicrobial susceptibility in samples from adult patients with chronic wounds. Methods: Patients treated at National Institute of Wounds Foundation between May and July 2014, with chronic ulcers in lower extremities with clinical inflammatory signs were recluted. Samples were cultured in aerobic and anaerobic atmosphere and species identification was performed by API (Biomerieux) galleries. The in vitro susceptibility was evaluated according to the Kirby Bauer method. Results: From 73 patients, 46 had infected wounds most of them were venous ulcers (33) with prevalence in polymicrobial infections and 10 with foot-diabetes ulcers with prevalence in monomicrobial infections (p ≤ 0.05). Sixty-eight strains were isolated and Enterobacteriaceae were predominant in monomicrobial infection (p ≤ 0.05) and the other groups were slightly higher in polymicrobial infection. The main species were Staphylococcus aureus (24%) followed by P. aeruginosa (18%). Fifty strains (77%) were resistant or multi-resistance. We emphasize resistance of S. aureus to ciprofloxacin (50%) and cefoxitin (37.5%), thus identifying resistance to methicillin in the community (CA-SAMR), all of which are sensitive to cotrimoxazole. Enterobacteria showed sensitivity to amikacin (95.5%), P. aeruginosa showed resistance to ciprofloxacin (33.3%) with high sensitivity to gentamicin (91.7%) and amikacin (83.3%), while Acinetobacter spp showed resistance to ciprofloxacin and ceftazidime in 60%, with 100% sensitivity to imipenem. 50% Streptococcus β hemolytic showed resistance to clindamycin and penicillin. Conclusion: These data provide epidemiological information on chronic wound infections, representing support for diagnosis, treatment and management of this pathology.

Sepsis neonatal tardía nosocomial en una unidad de terapia intensiva: agentes etiológicos y localización más frecuente / Late onset neonatal sepsis in an intensive care neonatal unit: etiological agents and most frequent location

Resumen Introducción: La sepsis neonatal nosocomial (SNN) es una entidad frecuente en las unidades de cuidados intensivos, donde causa una gran morbimortalidad. La ubicación más frecuente es bacteriemia, seguido de neumonía asociada a ventilador mecánico y vía urinaria. Objetivo: Conocer la etiología y localización más frecuente de la infección en el SNN. Población, Material y Métodos: Estudio retrospectivo, de prevalencias de enero a diciembre de 2015, realizado en la Unidad de Cuidados Intensivos Neonatal de un hospital de alta complejidad. Fueron incluidos todos los neonatos. Resultados: Se incluyeron 70 pacientes, se analizaron 88 episodios de SNN. La localización más frecuente fue sangre 40% de los casos, seguido de orina y aspirado traqueal en 25% respectivamente. Los microorganismos más frecuentemente aislados fueron Staphylococcus de diferentes tipos, seguido de Acinetobacter baumannii multi-resistente. La afectación del SNC fue de 32%. La mortalidad fue de 34%, elevándose a 50% ante un segundo episodio de SNN. La terapia empírica de elección fue vancomicina y carbapenem, ajustándose a antibiograma. Conclusiones: La infección más frecuente fue la bacteremia, principalmente por Staphylococcus resistentes a meticilina. La afectación del SNC fue elevada, lo mismo que la mortalidad.

Introduction: Nosocomial neonatal sepsis (NNS) is a frequent entity in intensive care units, causing great morbidity and mortality. The most frequent site is blood, followed by lungs and urine. Objective: To know the etiology and most frequent localization of infection in the NNS. Population, Material and Methods: Cross sectional study, from January to December 2015, performed in a teaching hospital. All newborns infants were included. Results: 70 patients were included, 88 episodes of NNS were analyzed. The most frequent localization was bacteremia in 40% of cases, followed by urinary tract infection and VAP in 25% respectively. The bacteria most frequently isolated were staphylococci of different types, followed by multiresistant Acinetobacter. The CNS involvement was 32%. Mortality was 34%, rising up to 50% with a second episode of NNS. The empirical therapy of choice was vancomycin and carbapenem, adjusting to antibiogram. Conclusions: The most frequent infection was bacteremia, mainly by staphylococci resistant to methicillin. CNS involvement was elevated, as well as mortality.