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Coagulase-negative Staphylococcus (CoNS) species inhibiting Staphylococcus aureus has been described in the skin of atopic dermatitis (AD) patients. This study evaluated whether Staphylococcus spp. from the skin and nares of AD and non-AD children produced antimicrobial substances (AMS). AMS production was screened by an overlay method and tested against NaOH, proteases and 30 indicator strains. Clonality was assessed by pulsed-field gel electrophoresis. Proteinaceous AMS-producers were investigated for autoimmunity by the overlay method and presence of bacteriocin genes by polymerase chain reaction. Two AMS-producers had their genome screened for AMS genes. A methicillin-resistant S. aureus (MRSA) produced proteinaceous AMS that inhibited 51.7% of the staphylococcal indicator strains, and it was active against 60% of the colonies selected from the AD child where it was isolated. On the other hand, 57 (8.8%) CoNS from the nares and skin of AD and non-AD children, most of them S. epidermidis (45.6%), reduced the growth of S. aureus and other CoNS species. Bacteriocin-related genes were detected in the genomes of AMS-producers. AMS production by CoNS inhibited S. aureus and other skin microbiota species from children with AD. Furthermore, an MRSA colonizing a child with AD produced AMS, reinforcing its contribution to dysbiosis and disease severity.

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Methicillin-resistant Staphylococcus aureus (MRSA) constitutes an important cause for concern in the field of public health, and the role of the food chain in the transmission of this pathogen and in antimicrobial resistance (AMR) has not yet been defined. The objectives of this work were to isolate and characterize coagulase-positive Staphylococcus (CoPS) and coagulase-negative Staphylococcus (CoNS), particularly S. aureus, from school dining rooms located in Argentina. From 95 samples that were obtained from handlers, inert surfaces, food, and air in 10 establishments, 30 Staphylococcus strains were isolated. Four isolates were S. aureus, and the remaining ones (N = 26) belonged to 11 coagulase-negative species (CoNS). The isolates were tested for susceptibility to nine antibiotics. The presence of genes encoding toxins (luk-PV, sea, seb, sec, sed, and see), adhesins (icaA, icaD), and genes that confer resistance to methicillin (mecA) and vancomycin (vanA) was investigated. The resistance rates measured for penicillin, cefoxitin, gentamicin, vancomycin, erythromycin, clindamycin, levofloxacin, trimethoprim-sulfamethoxazole, and tetracycline were 73%, 30%, 13%, 3%, 33%, 17%, 13%, 7%, and 7% of the isolates, respectively. Seventeen AMR profiles were detected, and 11 isolates were multidrug resistant (MDR). Seven methicillin-resistant Staphylococcus isolates were detected in the hands of handlers from four establishments, two of them were MRSA. Two S. aureus isolates presented icaA and icaD, another one, only icaD. The gene vanA was found in two isolates. In relation to S. aureus, resistance to vancomycin but not to gentamicin was detected. School feeding plays a key role in the nutrition of children, and the consumption of food contaminated with MRSA and vancomycin-resistant S. aureus (VRSA) can be a serious threat to health. In particular, it was detected that the handlers were the source of MRSA, VRSA, MR-CoNS (methicillin-resistant coagulase-negative Staphylococcus), and MDR isolates. The results obtained indicate that the vigilance of this pathogen in school dining rooms should be extreme.

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ABSTRACT Mastitis is one of the most important illnesses in specialized dairy herds worldwide due to the effects on production and animal health. The types caused by CNS has a special importance in a production where the main pathogens are controlled. The objective of the present work is to determine the prevalence of CNS in a dairy herd in Boyaca and also quantify the effects of every species of CNS in SCC. 40 cows were selected and sampled during 6 months, CMT was performed, and results from 1 to trace were sampled. The routine bacteriological test was also performed for CNS identification, and the isolating of CNS was performed through rpoB gene identification and through the type of strain using the pulse gel electrophoresis procedure. Out of 960 samples, 619 were positive for CNS growth. The most prevalent species were Staphylococcus epidermidis, S. chromogenes, S. sciuri, S. simulaans, S. haemolyticus and S. capitis. The results that were found here are similar to the results observed in different parts of the world, which confirms that they are pathogens that must be constantly evaluated because they can go unnoticed in routine controls, especially in those farms where major pathogens are not a serious problem. The results determined in this study demonstrate that CNS generates a slight increase in somatic cells.

RESUMEN La mastitis es una de las enfermedades más importantes en los rebaños lecheros especializados alrededor de todo el mundo debido a los efectos sobre la producción y la salud animal. Los tipos ocasionados por estafilococos coagualasa negativo (ECN) tienen una importancia especial en una producción en la que los principales patógenos están controlados. El objetivo del presente trabajo es determinar la prevalência del ECN en un hato lechero en Boyacá y cuantificar los efectos de cada especie de ECN en el conteo de células somáticas (CCS). Se seleccionaron 40 vacas y se tomaron muestras durante 6 meses, se realizó california mastitis test (CMT) y se tomaron muestras de los resultados desde 1 hasta donde hubo trazas. También se realizó la prueba bacteriológica de rutina para la identificación del ECN y el aislamiento del ECN se realizó mediante la identificación del gen rpoB y del tipo de cepa, usando el procedimiento de electroforesis en gel de pulso. De 960 muestras, 619 fueron positivas para el crecimiento del ECN. Las especies más prevalentes fueron Staphylococcus epidermidis, S. chromogenes, S. sciuri, S. simulans, S. haemolyticus y S. capitis. Los resultados encontrados aquí son similares a resultados en diferentes partes del mundo, lo que confirma que son patógenos que deben ser evaluados constantemente porque pueden pasar desapercibidos en los controles de rutina, especialmente en aquellas fincas donde los patógenos mayores no son un problema grave. Los resultados determinados en este estudio demuestran que el SNC genera un ligero aumento de células somáticas.

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Resumen Introducción: Vancomicina es un antimicrobiano ampliamente utilizado para infecciones por Staphylococcus coagulasa negativa en neonatos; sin embargo, no existe claridad sobre la dosis empírica que asegure su eficacia terapéutica. Objetivo: Evaluar la relación entre las dosis iniciales de vancomicina utilizadas en una Unidad de Cuidado Intensivo Neonatal (UCIN) con la eventualidad de alcanzar el objetivo terapéutico de área bajo la curva sobre concentración inhibitoria mínima (ABC/CIM) mayor a 400 µg/h/mL. Materiales y Método: Estudio descriptivo y retrospectivo, realizado entre febrero 2016 y marzo 2018. Se incluyeron neonatos en tratamiento con vancomicina por sospecha/confirmación de infección por cocáceas grampositivas y medición de concentraciones plasmáticas de vancomicina al inicio del tratamiento. La probabilidad de alcanzar el objetivo terapéutico se evaluó mediante re-muestreo de valores de ABC y CIM. Resultados: Se incluyeron 38 pacientes con 49 concentraciones plasmáticas de vancomicina. Los aislados microbiológicos se confirmaron en 94,7% de los pacientes (n = 36). Los valores de ABC/CIM en dos grupos (según niveles valle de vancomicina < 10 µg/mL y ≥ 10 µg/mL), fueron de una mediana de 327 (IQ 25-75 = 174-395) y 494 (IQ 25-75 = 318-631), respectivamente (p = 0,035). Las dosis empíricas utilizadas logran logran un objetivo terapéutico (ABC/CIM > 400) de sólo 47,7% considerando CIMs en nuestra institución. Conclusiones: Teniendo en cuenta las sensibilidades institucionales, no es posible asegurar alcanzar ABC/CIM > 400 µg/h/mL. Se debe seguir investigando para replantear las actuales estrategias de dosificación y así determinar la más apropiada para neonatos.

Abstract Background: Vancomycin is used for treating coagulase-negative staphylococcus infections in neonates. However, concerns about the appropriate empirical dosing required for optimal efficacy, still remain. Aim: To assess the relationship between the initial doses of vancomycin used in a Neonatal Intensive Care Unit (NICU) with the possibility of achieving therapeutic target of AUC024h/MIC > 400 µg/h/mL. Methods: Retrospective and descriptive study carried out between February 2016 and March 2018. All neonates treated with vancomycin for suspected/proven Gram-positive infection and with at least one trough serum concentration level were included. Probability of target attainment (PTA) was evaluated through resampling of AUC and MIC values. Results: Final dataset included 38 patients and 49 trough vancomycin levels; 94.7% of these cases (n = 36) were confirmed Gram-positive infections. The median AUC/MIC values for the trough values vancomycin < 10 µg/mL group and for the ≥ 10 µg/mL group were 327 (IQR 174-395) and 494 (IQR 318-631) respectively (p = 0.035). Current empirical dosing strategy has a 47.7% PTA (AUC/MIC > 400) when taking institutional MICs into account. Conclusions: It is not possible to assure achieving a AUC/MIC > 400 µg/h/mL when considering institutional sensibilities. Current empiric dosing strategies should be reconsidered and further investigation needs to be done to help determine the appropriate empirical dosing required for optimal efficacy in neonates.

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Abstract Objectives To present current evidence on the etiology, risk factors, diagnosis, and management of early and late neonatal sepsis. Source of data Non-systematic review of the Medline (PubMed), Scopus, Web of Science, Cochrane, and Google Scholar databases regarding the following terms: neonatal sepsis, early neonatal sepsis, late neonatal sepsis, empirical antibiotic therapy, sepsis calculator, vancomycin, newborn, preterm newborn. Data synthesis Neonatal sepsis is a frequent cause of neonatal morbidity and mortality. Its diagnosis is difficult. Continuous observation of the patient is critical to diagnostic suspicion. When neonatal sepsis is suspected, bacteriological tests should be collected. Vancomycin should not be routinely using in the empirical antibiotic regimen in late neonatal sepsis, and the main protective mechanisms against neonatal sepsis are handwashing and the use of breast milk. Conclusions Newborns constitute a group that is more vulnerable to sepsis. Knowledge of risk factors and etiological agents allows a better approach to the newborn with sepsis.

Resumo Objetivos Apresentar evidências atuais na etiologia, fatores de risco, diagnóstico e manejo da sepse neonatal precoce e tardia. Fontes de dados Revisão não sistemática feita nas bases de dados Medline (PubMed), Scopus, Web of Science, Cochrane, Google Scholar sobre os temas sepse neonatal, sepse neonatal precoce, sepse neonatal tardia, antibioticoterapia empírica, sepsis calculator, vancomicina, recém-nascido, recém-nascido pré-termo. Síntese de dados A sepse neonatal é uma causa frequente de morbimortalidade neonatal. O seu diagnóstico é difícil. A observação contínua do paciente é fundamental para uma suspeição diagnóstica. Ao se suspeitar de sepse neonatal devem-se coletar exames bacteriológicos. Não usar, rotineiramente, vancomicina no esquema empírico de antibiótico na sepse neonatal tardia. Os principais mecanismos protetores da sepse neonatal são a lavagem de mãos e o uso do leite materno. Conclusões Os recém-nascidos constituem um grupo mais vulnerável à sepse. O conhecimento dos fatores de risco e dos agentes etiológicos permite uma melhor abordagem do recém-nascido séptico.

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OBJECTIVES: To present current evidence on the etiology, risk factors, diagnosis, and management of early and late neonatal sepsis. SOURCE OF DATA: Non-systematic review of the Medline (PubMed), Scopus, Web of Science, Cochrane, and Google Scholar databases regarding the following terms: neonatal sepsis, early neonatal sepsis, late neonatal sepsis, empirical antibiotic therapy, sepsis calculator, vancomycin, newborn, preterm newborn. DATA SYNTHESIS: Neonatal sepsis is a frequent cause of neonatal morbidity and mortality. Its diagnosis is difficult. Continuous observation of the patient is critical to diagnostic suspicion. When neonatal sepsis is suspected, bacteriological tests should be collected. Vancomycin should not be routinely using in the empirical antibiotic regimen in late neonatal sepsis, and the main protective mechanisms against neonatal sepsis are handwashing and the use of breast milk. CONCLUSIONS: Newborns constitute a group that is more vulnerable to sepsis. Knowledge of risk factors and etiological agents allows a better approach to the newborn with sepsis.

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Abstract INTRODUCTION: Contaminated hospital environments contribute to the transmission of microorganisms associated with healthcare. Contaminated surfaces handled by patients or healthcare professionals are a source of microorganism transmission by hand. Methicillin-resistant Staphylococcus bacteria are among the main agents responsible for increasing healthcare-associated infections in Brazil and worldwide. METHODS: The objective of this study was to screen and characterize methicillin-resistant Staphylococcus spp. on surfaces near patients in an intensive care unit. Microbiological samples, collected from ten beds in an intensive care unit with five sampling sites, were inoculated into a methicillin-resistant Staphylococcus aureus chromogenic medium. MALDI-TOF and PCR analyses were used to identify the bacteria. Antimicrobial susceptibility was determined using the disk diffusion test. The presence of the mecA gene was investigated using PCR. RESULTS: We observed that 44 out of the 50 sampling sites presented grown isolates in the methicillin-resistant Staphylococcus aureus medium. The incidence of isolated microorganisms on the right side rail, left side rail, tables, infusion pump keypad, and cardiac monitor were 18.8 %, 36.7 %, 10.9 %, 2.4 %, and 31 %, respectively. The 42 isolates included in this study were identified as coagulase-negative Staphylococcus. All of these microorganisms were multidrug-resistant and mecA gene-positive. CONCLUSIONS: This study identified the presence of methicillin-resistant coagulase-negative Staphylococcus on the beds of an intensive care unit, providing evidence for the necessity of assertive actions to decrease the risk of healthcare-associated infections at the site.

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The antibiotic resistance among coagulase - negative Staphylococcus (CoNS) species towards methicillin is rarely reported in veterinary medicine. Under the aspect/concept of One Health, those strains pose a risk to human health due to the presence in canine pets where the transfer of resistant genetic markers might occur to other staphylococci species. The aim of this study was to investigate the antimicrobial resistance pattern among Coagulase Negative Staphylococci (CoNS) isolated from asymptomatic dogs and those affected by topic infections. Swabs from 254 dogs were first seeded in Mannitol Salt Agar. Species identification was conducted by Matrix Assisted Laser Desorption ionization - time of flight (MALDI-TOF ms) as previously described. The susceptibility test was performed by disk diffusion according to CLSI standards. Detection of mecA gene was performed. CoNS could be recovered from both groups of dogs and an alarming presence of methicillin-resistant coagulase negative staphylococci (MRCoNS) was confirmed, in 10.2% (17/166) of the samples. Eight of those methicillin resistant strains were isolated from asymptomatic dogs whereas nine were present in dogs affected by pyoderma and otitis externa.

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Infecções intramamárias por Staphylococcus coagulase negativa (SCN) têm sido relatadas em muitos sistemas de produção leiteira no mundo todo. A qualidade do leite é definida com base na sua composição e higiene. A contagem de células somáticas (CCS) é considerada como o principal indicador de qualidade do leite e da situação da mastite nos rebanhos. Objetivou-se assim, estudar as consequências da mastite por SCN sobre a qualidade do leite através de um estudo de meta-análise. Concluiu-se que os SCN são patógenos emergentes e que vêm causando graves consequências sobre a qualidade do leite.

Coagulase negative Staphylococcus (CNS) intramammary infections have been reported in many dairy production systems worldwide. Milk quality is defined based on its composition and hygiene. Somatic Cell Count (CCS) is considered the main indicator of milk quality and of the mastitis situation in herds. The aim of this study was to investigate the effects of CNS mastitis on milk quality through a meta-analysis study. In this context, the research pointed out that CNS are emerging pathogens and have been causing serious consequences on milk quality.

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Infecções intramamárias por Staphylococcus coagulase negativa (SCN) têm sido relatadas em muitos sistemas de produção leiteira no mundo todo. A qualidade do leite é definida com base na sua composição e higiene. A contagem de células somáticas (CCS) é considerada como o principal indicador de qualidade do leite e da situação da mastite nos rebanhos. Objetivou-se assim, estudar as consequências da mastite por SCN sobre a qualidade do leite através de um estudo de meta-análise. Concluiu-se que os SCN são patógenos emergentes e que vêm causando graves consequências sobre a qualidade do leite.(AU)

Coagulase negative Staphylococcus (CNS) intramammary infections have been reported in many dairy production systems worldwide. Milk quality is defined based on its composition and hygiene. Somatic Cell Count (CCS) is considered the main indicator of milk quality and of the mastitis situation in herds. The aim of this study was to investigate the effects of CNS mastitis on milk quality through a meta-analysis study. In this context, the research pointed out that CNS are emerging pathogens and have been causing serious consequences on milk quality.(AU)

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Coagulase-negative staphylococci (CNS) are important pathogens causing nosocomial infections worldwide with increasing resistance to antimicrobials. The aim of this study was to characterize resistance aspects of CNS isolated from patients with bloodstream infections acquired in hospitals in Belo Horizonte, MG, Brazil. Staphylococcus strains were characterized using repetitive sequence-based polymerase chain reaction (rep-PCR) fingerprinting with (GTG)5 primer. Phenotypic resistance was analyzed using AST-P5085 card (bioMérieuxVitek®). PCR was used to detect mecA, vanA, blaZ, ermA/B/C, aac-aphD, and SCC-mec. For statistical analyses, we used hierarchical cluster, chi-square test (χ2), and correspondence. Several clusters were formed within the same species using (GTG)5 primer, and strains showed resistance to the following antimicrobials: benzylpenicillin (100%); oxacillin (93.1%); gentamicin (36.3%); ciprofloxacin (63.7%); moxifloxacin (32.7%); norfloxacin (81.0%); erythromycin (86.2%); clindamycin (75.8%); linezolid, teicoplanin and vancomycin (1.7%); tigecycline (0%); fusidic acid (10.35%); rifampicin (13.7%); and trimethoprim/sulfamethoxazole (46.5%). Regarding genotypic analyses, 40%, 0%, 78%, 42%, 100%, 24%, and 30% were positive for mecA, vanA, blaZ, ermA, ermB, ermC, and aac-aphD, respectively. Regarding staphylococcal cassette mec (SCCmec) type, 3.4% presented type I; 5.0% type II; 27.1% type III; 20.3% type IIIA; and 32.2% type IIIB. Six clusters were formed and frequency distributions of resistant strains to oxacillin, gentamicin, ciprofloxacin, moxifloxacin, norfloxacin, erythromycin, clindamycin, linezolid, teicoplanin, vancomycin, fusidic acid, rifampicin, and trimethoprim/sulfamethoxazole, and mecA, blaZ, ermC, aac-aphD, and SCCmec type differed (p < 0.001). In conclusion, the strains investigated in this study were multidrug resistant and carried multiple antibiotic resistance genes.

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Un total de 79 cepas de S. aureus y 47 cepas de Staphylococcus coagulasa negativa (SCN) aislados de muestras de leche de vaca con mastitis subclínica fueron evaluadas para establecer su propiedad para formar biopelícula como uno de los factores de virulencia más importantes. Usando el método de Rojo Congo Agar, 80% de las cepas de S. aureus fueron productores de limo, mientras que en las cepas de SCN el porcentaje fue de 32%. Por el método de microplaca, 55%, 17% y 28% de los aislamientos de S. aureus fueron fuerte, moderadas y débiles productoras de biopelícula, mientras en los SCN el porcentaje fue 43%, 17% y 40%, respectivamente. Se realizó un ensayo de Reacción en Cadena de la Polimerasa (PCR) a todos los aislamientos con la finalidad de identificar el gen A de adhesión intracelular (icaA). En las cepas de S. aureus el gen icaA estuvo presente en el 65% de los aislamientos, y en los SCN en el 11%. La mayoría de las cepas de S. aureus caracterizados en el estudio fueron formadores de biopelícula, lo cual sugiere que está tiene un importante papel en la virulencia de S. aureus aislados de infecciones intramamarias en bovinos del estado Zulia.

A total of 79 S. aureus strains and 47 coagulase negative Staphylococcus (CNS) isolates from cow milk suffering subclinical mastitis were investigated for their ability to form biofilm as one of the most important virulence factors. Using Congo Red Agar method, 80% of S. aureus strains were slime producers, while in CNS was 32%. By microtiter plate method, 55%, 17%, and 28% of S. aureus isolates were strong, moderate, and weak biofilm producers, respectively, while in CNS the percentages were 43%, 17%, and 40%, respectively. All isolates were screened by Polymerase chain reaction (PCR) for amplification of intercellular adhesion gene A (icaA). In S. aureus isolates the icaA gene was present in 65 % while in CNS was 11%. The majority of S. aureus characterized in this study formed biofilm, which suggests that biofilm formation has an important role in the virulence of S. aureus isolated from bovine intramammary infections in Zulia state.

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In addition to Staphylococcus aureus nowadays other coagulase-positive staphylococci (CoPS) and coagulase-negative staphylococci (CoNS), earlier considered of minor importance, are now accepted as relevant pathogens for humans and animals. The involvement of these microorganisms in bovine mastitis etiology and the possibility their transmission through milk to humans justify the requirement of developing reliable methods for identification of the most frequent species among them. The purpose of this study was to compare the phenotypic techniques with the genotypic method carried out by sequencing of the rpoB gene in identification of several species of the genus Staphylococcus isolated from bovine mastitis. A total of 300 staphylococci isolates of bovine mastitis cases from several Brazilian dairy herds were studied by phenotypic and genotypic techniques, respectively: 150 CoPS and 150 CoNS strains. A total of 18 CoNS different species and 4 CoPS species were identified. Among the CoNS the following species were recognized: 48 (32%) Staphylococcus warneri, 22(15%) S. epidermidis, 20(13%) S. hyicus, 10(7%) S. xylosus, 7(5%) S. haemolyticus, 6(4%) S. simulans, 6(4%) S. schleiferi subsp schleiferi, 6(4%) S. hominis, 5(3%) S. pasteuri, 4(2.7%) S. cohnii, 3(2%) S. saprophyticus subsp. saprophyticus 3(2%) S. chromogenes 3(2%) S. sciuri, 2(1%) S. saccharolyticus, 2(1%) S. lugdunensi, 1(0,7%) S. auricularis, 1(70%) S. saprophyticus subsp. bovis, 1(0.7%) S. capitis. And among the 150 CoPS were identified respectively: 105 (70%) S. aureus, 21(14%), S. hyicus, 19(13%) S. intermedius e 5(3%) S. schleiferi subsp coagulans. Considering the 150 CoNS isolates, the identifications performed by phenotypic and genotypic tests presented 96.7% of concordance, kappa coefficient of agreement = 0.933, SE (standard error) of kappa=0.021 (95% confidence interval: 0.893 to 0.974), Pearsons correlation coefficient (r) = 0.9977, (confidence interval 95%: 0.9938 a 0.9992) and [...](AU)

Além de Staphylococcus aureus atualmente outros estafilococos coagulase positiva (SCP) e estafilococos coagulase-negativos (SCN), anteriormente considerados de menor relevância, são reconhecidos como importantes patógenos para humanos e animais. O envolvimento desses micro-organismos na etiologia da mastite bovina e a possibilidade da sua transmissão através do leite aos humanos justifica a utilização de métodos confiáveis para a identificação das espécies mais frequentes. O objetivo deste estudo foi comparar as técnicas fenotípicas com o método genotípico realizada por sequenciamento do gene rpoB na identificação de espécies do gênero Staphylococcus spp. isolados de mastite bovina. Um total de 300 estafilococos isolados de casos de mastite bovina em diferentes rebanhos leiteiros brasileiros foram estudados por técnicas fenotípicas e genotípicas, respectivamente: 150 linhagens de SCP e 150 linhagens de SCN. Foram identificados um total de 18 espécies de SCN e 4 espécies SCP. Entre os SCN as seguintes espécies identificadas: 48 (32%) Staphylococcus warneri, 22 (15%) S. epidermidis, 20 (13%) S. hyicus, 10 (7%) S. xylosus, 7 (5%) S. haemolyticus, 6 (4%) S. simulans, 6 (4%) S. schleiferi subsp schleiferi, 6 (4%) S. hominis, 5 (3%) S. pasteuri, 4 (2,7%) S. cohnii, 3 (2%) S. saprophyticus subsp. saprophyticus, 3 (2%) S. chromogenes, 3 (2%) S. sciuri, 2 (1%) S. saccharolyticus, 2 (1%) S. lugdunensi, 1 (0,7%) S. auricularis, 1 (70 %) S. saprophyticus subsp. bovis, 1 (0,7%) S. capitis. E entre as 150 SCP foram identificados, 105 (70%) S. aureus, 21 (14%), S. hyicus, 19 (13%) S. intermedius e 5 (3%) S. schleiferi subsp coagulans. Considerando-se os 150 SCN isolados, as identificações realizadas por testes fenotípicos e genotípicos apresentaram 96,7% de concordância, coeficiente de concordância kappa = 0,933, SE (erro padrão) de [...](AU)

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In addition to Staphylococcus aureus nowadays other coagulase-positive staphylococci (CoPS) and coagulase-negative staphylococci (CoNS), earlier considered of minor importance, are now accepted as relevant pathogens for humans and animals. The involvement of these microorganisms in bovine mastitis etiology and the possibility their transmission through milk to humans justify the requirement of developing reliable methods for identification of the most frequent species among them. The purpose of this study was to compare the phenotypic techniques with the genotypic method carried out by sequencing of the rpoB gene in identification of several species of the genus Staphylococcus isolated from bovine mastitis. A total of 300 staphylococci isolates of bovine mastitis cases from several Brazilian dairy herds were studied by phenotypic and genotypic techniques, respectively: 150 CoPS and 150 CoNS strains. A total of 18 CoNS different species and 4 CoPS species were identified. Among the CoNS the following species were recognized: 48 (32%) Staphylococcus warneri, 22(15%) S. epidermidis, 20(13%) S. hyicus, 10(7%) S. xylosus, 7(5%) S. haemolyticus, 6(4%) S. simulans, 6(4%) S. schleiferi subsp schleiferi, 6(4%) S. hominis, 5(3%) S. pasteuri, 4(2.7%) S. cohnii, 3(2%) S. saprophyticus subsp. saprophyticus 3(2%) S. chromogenes 3(2%) S. sciuri, 2(1%) S. saccharolyticus, 2(1%) S. lugdunensi, 1(0,7%) S. auricularis, 1(70%) S. saprophyticus subsp. bovis, 1(0.7%) S. capitis. And among the 150 CoPS were identified respectively: 105 (70%) S. aureus, 21(14%), S. hyicus, 19(13%) S. intermedius e 5(3%) S. schleiferi subsp coagulans. Considering the 150 CoNS isolates, the identifications performed by phenotypic and genotypic tests presented 96.7% of concordance, kappa coefficient of agreement = 0.933, SE (standard error) of kappa=0.021 (95% confidence interval: 0.893 to 0.974), Pearson's correlation coefficient (r) = 0.9977, (confidence interval 95%: 0.9938 a 0.9992) and in relation to 150 CPS isolates it was detected an agreement of 98.7%, kappa = 0.960, SE of kappa = 0.016, (95% confidence interval: 0.929 to 0.992) Pearson's correlation coefficient (r) = 0.9994 (95% confidence interval: 0.9681 to 1.0000). The verified agreement strength between the identification methods can be considered as excellent. These results assure that according to laboratory resources any of them will be suitable to perform the staphylococci identification.(AU)

Além de Staphylococcus aureus atualmente outros estafilococos coagulase positiva (SCP) e estafilococos coagulase-negativos (SCN), anteriormente considerados de menor relevância, são reconhecidos como importantes patógenos para humanos e animais. O envolvimento desses micro-organismos na etiologia da mastite bovina e a possibilidade da sua transmissão através do leite aos humanos justifica a utilização de métodos confiáveis para a identificação das espécies mais frequentes. O objetivo deste estudo foi comparar as técnicas fenotípicas com o método genotípico realizada por sequenciamento do gene rpoB na identificação de espécies do gênero Staphylococcus spp. isolados de mastite bovina. Um total de 300 estafilococos isolados de casos de mastite bovina em diferentes rebanhos leiteiros brasileiros foram estudados por técnicas fenotípicas e genotípicas, respectivamente: 150 linhagens de SCP e 150 linhagens de SCN. Foram identificados um total de 18 espécies de SCN e 4 espécies SCP. Entre os SCN as seguintes espécies identificadas: 48 (32%) Staphylococcus warneri, 22 (15%) S. epidermidis, 20 (13%) S. hyicus, 10 (7%) S. xylosus, 7 (5%) S. haemolyticus, 6 (4%) S. simulans, 6 (4%) S. schleiferi subsp schleiferi, 6 (4%) S. hominis, 5 (3%) S. pasteuri, 4 (2,7%) S. cohnii, 3 (2%) S. saprophyticus subsp. saprophyticus, 3 (2%) S. chromogenes, 3 (2%) S. sciuri, 2 (1%) S. saccharolyticus, 2 (1%) S. lugdunensi, 1 (0,7%) S. auricularis, 1 (70 %) S. saprophyticus subsp. bovis, 1 (0,7%) S. capitis. E entre as 150 SCP foram identificados, 105 (70%) S. aureus, 21 (14%), S. hyicus, 19 (13%) S. intermedius e 5 (3%) S. schleiferi subsp coagulans. Considerando-se os 150 SCN isolados, as identificações realizadas por testes fenotípicos e genotípicos apresentaram 96,7% de concordância, coeficiente de concordância kappa = 0,933, SE (erro padrão) de kappa = 0,021 (95% intervalo de confiança: 0,893-0,974), coeficiente de correlação de Pearson (r) = 0,9977, (intervalo de confiança de 95%: 0,9938 a 0,9992) e em relação a 150 SCP isolados foi observado uma concordância de 98,7%, kappa = 0,960, sE de kappa = 0,016, (95% de intervalo de confiança: 0,929 a 0,992) coeficiente de correlação de Pearson (r) = 0,9994 (95% intervalo de confiança: 0,9681-1,0000). A correlação entre os métodos de identificação pode ser considerada como excelente. Esses resultados demonstraram que de acordo com os recursos disponíveis no laboratório, poderia ser utilizada qualquer uma das metodologias.(AU)

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La resistencia a los antimicrobianos en bacterias Gram positivas como Staphylococcus coagulasa negativa constituye una amenaza mundial emergente. El propósito de la presente investigación fue identificar los genes de resistencia a oxacilina (mecA), eritromicina (erm y msrA), y gentamicina aac(6´)/aph(2´´), en cepas de Staphylococcus coagulasa negativa aisladas de hemocultivos de pacientes atendidos en el Hospital Universitario de Maracaibo. La detección fenotípica se realizó mediante métodos automatizados. Se utilizó la reacción en cadena de la polimerasa para detectar la presencia de genes de resistencia a los antimicrobianos. Se estudiaron 34 cepas cuya distribución por especie fue: S. haemolyticus (38,23%), S. epidermidis (29,42%), S. hominis (26,47%), S. xylosus y S. capitis (5,88% cada uno). Todas las cepas fueron resistentes a oxacilina. La resistencia a gentamicina varió entre 38,46% y 100%; mientras que la resistencia a eritromicina osciló entre 77,78% y 100%. Los análisis mostraron la presencia de los genes mecA (100%), ermA (35,2%), ermC (41,17%), msrA (17,64%), y aac(6´)/aph(2´´) (61,76%). En conclusión, se encontró una alta frecuencia de genes de resistencia a estos antibióticos y la Unidad de Cuidados Intensivos fue el servicio médico donde se aisló el mayor porcentaje de cepas portadoras de estos genes.

Resistance to antimicrobials in Gram-positive bacteria such as coagulase negative Staphylococcus is an emerging global threat. The purpose of this research was to identify the genes for resistance to oxacillin (mecA), erythromycin (erm and msrA), and gentamicin aac(6´)/aph(2´´), in Staphylococcus coagulase negative strains isolated from blood cultures from patients attended at the University Hospital in Maracaibo. Phenotypic detection was performed using automated methods. Polymerase chain reaction was used for the detection of antimicrobial resistance genes. Be studied 34 strains whose distribution by species was: S. haemolyticus (38.23%), S. epidermidis (29.42%), S. hominis (26.47%), S. xylosus and S. capitis (5.88% each one). All strains were resistant to oxacillin. Gentamicin resistance varied between 38.46% and 100%; while the erythromycin resistance ranged between 77.78% and 100%. The analyses showed the presence of genes mecA (100%), ermA (35.2%), ermC (41.17%), msrA (17.64%), and aac(6´)/aph (2´´) (61,76%). In conclusion, is found a high frequency of genes for resistance to these antibiotics and the intensive care unit was the health service where the highest percentage of isolated strains carriers of these genes.

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Introducción: La bacteremia por Staphylococcus coagulasa negativo (SCN) es una infección que actualmente genera significativa morbimortalidad dado el tipo de pacientes a los que afecta y el cambio en el perfil de susceptibilidad a los antibióticos. Objetivo: Describir características de los pacientes con aislamientos de SCN en sangre con CIM para vancomicina ≥2. Metodología: Estudio observacional descriptivo en pacientes hospitalizados ≥ 15 años. Resultados: De 130 aislamientos, 38 (29,23%) tenían CIM para vancomicina ≥ 2. La mediana de edad fue de 54 años y la razón hombre:mujer, de 1,37:1. Las especies más frecuentes fueron: S. epidermidis (71,1%), S. haemolyticus (13,2%) y S. hominis (7,9%). El 44,7% tenían antecedentes de inmunosupresión, siendo las más importantes: neoplasias (28,9%), enfermedad reumatológica (5,2%) y VIH (2,6%). El 81,5% tenían accesos vasculares, el 97,3% antecedente de hospitalización previa y el 60,5% habían requerido manejo en la UCI. El 81,4% tuvieron exposición previa a antibióticos, y los más utilizados fueron: betalactámicos (78,9%) y vancomicina (50%). En el 2,6% se documentó endocarditis infecciosa. De los aislamientos de SCN con CIM para vancomicina ≥2 µg/ml, el 26,3% fueron sensibles a meticilina. Los principales tratamientos recibidos fueron: daptomicina (31,5%), vancomicina (21%), linezolid (15,7%) y betalactámicos (10,5%). Se utilizó terapia combinada en el 10,5%. La mortalidad general fue del 15,8%, y la mortalidad atribuible, del 33,3%. Conclusión: Un porcentaje considerable de aislamientos tenían heterorresistencia para vancomicina. La bacteremia estuvo asociada con accesos vasculares, hospitalizaciones previas, tratamientos en cuidado intensivo y exposición previa a antibióticos. La inmunosupresión es la comorbilidad más importante, y la mortalidad es significativa.

Introduction: Currently, coagulase-negative Staphylococcus (CNS) bacteremia is an infection that leads to significant morbidity and mortality given the type of patients affected and the recent changes in antimicrobial susceptibility. Objective: To describe the characteristics of patients with CNS blood isolates with vancomycin MIC ≥ 2. Methodology: Descriptive observational study on hospitalised patients ≥ 2 15 years of age. Results: Of 130 isolates, 38 (29,23%) contained vancomycin MIC ≥ 2. The median age was 54 years and the male:female ratio was 1.37:1. The most frequent species were S. epidermidis (71.1%), S. haemolyticus (13.2%) and S. hominis (7.9%). Some 44.7% of patients had a history of immunosuppression, including: neoplasms (28.9%), rheumatologic disease (5.2%) and HIV (2.6%). Some 81,5% had vascular access; 97.3% had previous hospitalisations and 60.5% had required intensive care. A total of 81.4% of the patients had prior exposure to antibiotics and the most commonly used were beta-lactams (78.9%) and vancomycin (50%). Infective endocarditis was documented in 2.6%. Of the CNS isolates with vancomycin MIC ≥ 2, 26.3% were sensitive to methicillin. The main treatments received were: daptomycin (31.5%), vancomycin (21%), linezolid (15.7%) and betalactams (10.5%). Combined therapy was performed in 10.5%. The overall mortality was 15.8% and attributable mortality was 33.3%. Conclusion: A significant proportion of isolates were hetero-resistant to vancomycin. The bacteremia was associated with vascular access, previous hospitalisations, intensive care treatments and prior antibiotic exposure. Immunosuppression is the most important comorbidity and mortality is significant.

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Consumption of goat milk has been increasing due to its nutritional characteristics and health benefits. Therefore, assessment of the presence of foodborne pathogens in this product is critical to ensure its safety to consumers. The present study aimed to identify common foodborne pathogens in raw goat milk. Fifty-three samples of raw goat milk from 11 farms were collected and cultured for the presence of Salmonella spp. and Listeria monocytogenes, as well as for enumeration and isolation of coagulase-positive and coagulase-negative Staphylococcus (CPS and CNS, respectively). All samples tested negative for Salmonella spp. and L. monocytogenes. The CPS counts in raw goat milk samples were predominantly less than 2 log cfu/mL (n=39), and CNS counts were predominantly higher than 3 log cfu/mL (n=42). Based on Staphylococcus counts, 51 isolates were selected (CPS=26; CNS=25) and tested by PCR for the presence of classic enterotoxin-encoding genes (sea, seb, sec, sed, and see). Only 3 isolates (CPS=2, CNS=1) were negative for all enterotoxin-encoding genes tested, and the genotype sec and see was the most frequent (n=16), followed by sea, sec, and see (n=13) and sec (n=13); sed was not detected in any isolate. The frequencies of enterotoxin-encoding genes for CPS and CNS were similar, demonstrating the equivalence of both groups in harboring these virulent markers. These results suggest that Salmonella and L. monocytogenes are not frequent contaminants of raw goat milk, but that Staphylococcus spp. that are capable of producing enterotoxins are prevalent; therefore, consumers of raw goat milk and products made from raw milk are at risk.

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Introducción: La sepsis es causa importante de morbimortalidad neonatal. Objetivos: Detectar el tiempo en que la curva de crecimiento bacteriano es evidenciada en la muestra de sangre inoculada en los hemocultivos y comparar estos tiempos de crecimiento bacteriano entre bacterias gramnegativas y grampositivas, entre los tipos de sepsis neonatal y determinar las bacterias más frecuentemente aisladas entre neonatos prematuros y de término. Pacientes y método: Estudio descriptivo de recién nacidos en riesgo de sepsis o con sospecha de sepsis por manifestaciones clínicas o de laboratorio, en que se evaluaron 114 hemocultivos positivos entre 1.932 hemocultivos tomados entre mayo de 2010 y mayo de 2014. Los datos se analizaron con Stata® 11.0. Resultados: El 5,9% de los hemocultivos tuvieron crecimiento bacteriano. La mediana y rango intercuartílico de tiempos de crecimiento bacteriano para gramnegativos fue 11 h (10-13 h), para grampositivos diferentes a Staphylococcus coagulasa negativo (SCoN) 12 h (12-18 h) y para SCoN 42h (36-44h). El 95,8% de las bacterias grampositivas y el 96% de las gramnegativas tuvieron tiempos de crecimiento bacteriano ≤ 24 h de incubación, mientras que en los SCoN el 100% de los hemocultivos fue positivo en ≤ 62 h de incubación. Conclusión: El 100% de sepsis por bacterias gramnegativas, grampositivas no SCoN y 90% de las ocasionadas por SCoN, son identificadas en los hemocultivos en las primeras 48 h, por lo cual podemos concluir que para descartar una sepsis, un período de incubación en hemocultivos de 48 h es suficiente.

Introduction: Sepsis is a major cause of neonatal morbidity and mortality. Objectives: To detect the time when the bacterial growth curve is evidenced in the blood sample inoculated blood cultures and comparing the times of bacterial growth between Gram negative and Gram positive bacteria, among the types of neonatal sepsis and identifying microorganisms more often isolated from preterm and term. Patients and method: A descriptive study. 114 positive blood cultures from 1,932 blood cultures taken from 01-May-2010 and 31-May-2014 were evaluated. Data were analyzed with Stata® 11.0. Results: 5.9% of blood cultures had bacterial growth. The median and interquartile range of Gram negative times of bacterial growth was 11 h (10-13 h), for Gram positive coagulase-negative Staphylococcus different (CoNS) 12h (12-18h) and CoNS 42h (36-44h). 95.8% of Gram positive and 96% of Gram negative, were the times of bacterial growth ≤ 24 h incubation, whereas the 100% CoNS was positive ≤ 62 h of incubation. Conclusion: 100% of sepsis by Gram negative and Gram positive no CoNS and 90% of those caused by CoNS are identified in blood cultures in 48 h, so we can conclude that to rule out sepsis, an incubation period of 48 h in blood cultures is sufficient.

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INTRODUCTION: Sepsis is a major cause of neonatal morbidity and mortality. OBJECTIVES: To detect the time when the bacterial growth curve is evidenced in the blood sample inoculated blood cultures and comparing the times of bacterial growth between Gram negative and Gram positive bacteria, among the types of neonatal sepsis and identifying microorganisms more often isolated from preterm and term. PATIENTS AND METHOD: A descriptive study. 114 positive blood cultures from 1,932 blood cultures taken from 01-May-2010 and 31-May-2014 were evaluated. Data were analyzed with Stata(®) 11.0. RESULTS: 5.9% of blood cultures had bacterial growth. The median and interquartile range of Gram negative times of bacterial growth was 11h (10-13h), for Gram positive coagulase-negative Staphylococcus different (CoNS) 12h (12-18h) and CoNS 42h (36-44h). 95.8% of Gram positive and 96% of Gram negative, were the times of bacterial growth≤24h incubation, whereas the 100% CoNS was positive≤62h of incubation. CONCLUSION: 100% of sepsis by Gram negative and Gram positive no CoNS and 90% of those caused by CoNS are identified in blood cultures in 48h, so we can conclude that to rule out sepsis, an incubation period of 48h in blood cultures is sufficient.

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BACKGROUND: Coagulase-negative Staphylococcus (CoNS) is commonly isolated from patients with chronic rhinosinusitis (CRS). However, the role of CoNS in CRS remains controversial. We aimed to determine the association between positive CoNS culture at functional endoscopic sinus surgery (FESS) and CRS severity. METHODS: Adult CRS patients who underwent FESS between October 1, 2007 to December 31, 2011 were recruited. Patient demographics, disease characteristics, medication use, Lund-Mackay computed tomography (CT) scores, and 22-item Sino-Nasal Outcome Test (SNOT-22) scores were collected at baseline before FESS. Intraoperative cultures were obtained in a standard manner. Patients were placed into 2 groups based on culture findings: patients with CoNS as the sole positive culture result and patients with all other positive culture results, including CoNS, as part of a polymicrobial culture. RESULTS: A total of 376 CRS patients met the criteria; 106 patients (28%) had CoNS as their only isolate, 260 (69%) had other positive cultures, and 10 (3%) had no bacterial growth. Compared to patients with other positive cultures, patients with the sole result of CoNS were significantly less likely to have a history of FESS (52% vs 65%, p = 0.019), nasal polyps (50% vs 65%, p = 0.006), and had a better Lund-Mackay CT score (11.95 vs 14.18, p = 0.020). After adjusting for all factors in the multiple logistic regression model, CoNS as the sole positive culture result was independently associated with having no history of FESS (odds ratio [OR] = 0.45; 95% confidence interval [CI], 0.22 to 0.94; p = 0.034). CONCLUSION: Positive intraoperative CoNS cultures alone do not result in increased CRS disease burden by objective or subjective measures as compared to patients with other bacterial or polymicrobial culture isolates.

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