Microbiological evaluation of pasty and soft ice cream / Avaliação microbiológica de sorvetes pastosos e expressos / Evaluación microbiológica de helados pastosos y suaves

Ice cream is susceptible to contamination by handling and bad hygiene conditions during both the storage process and the fractioning for sale, and once contaminated, it can cause diseases. The purpose of this survey was to evaluate the microbiological quality of ice cream sold in bulk, of pasty and soft types, offered for consuming. Thirty samples of pasty ice cream sold in bulk, and thirty samples of soft ice cream were analyzed through the counting of thermotolerant coliforms, coagulase-positive Staphylococcus spp., and searching for the presence of Salmonella spp. During the study, a total of ten (33%) samples of pasty ice cream and five (16%) samples of soft ice cream were found to be beyond the limits established by the Brazilian law. Salmonella spp. was found in four samples (6.7%). These results are an alert for the need of greater attention to the microbiological quality of ice cream in order to ensure the safety of its consumers.(AU)

Os sorvetes são suscetíveis à contaminação pela manipulação e más condições higiênicas durante o processamento, armazenamento e do fracionamento para venda, uma vez contaminados podem causar doenças. O objetivo deste estudo foi avaliar a qualidade microbiológica de sorvetes, vendidos a granel, pastosos e expressos, oferecidos para consumo. Trinta amostras de sorvete pastoso, vendido a granel, e trinta amostras de sorvete expresso foram analisadas realizando-se contagem de coliformes termotolerantes, Staphylococcus spp. coagulase-positiva e pesquisando-se a presença de Salmonella spp. Foram detectadas dez (33%) amostras de sorvete pastoso e cinco (16%) amostras de sorvete expresso fora dos limites estabelecidos pela legislação brasileira. Salmonella spp. foi encontrado em quatro amostras (6,7%). Esses resultados alertam para a necessidade de uma maior atenção à qualidade microbiológica dos sorvetes, a fim de garantir a segurança do consumidor.(AU)

Los helados son susceptibles a la contaminación por manipulación y malas condiciones higiénicas durante el procesamiento, almacenamiento y fraccionamiento para venta, una vez contaminados pueden causar enfermedades. El objetivo de este estudio ha sido evaluar la calidad microbiológica de helados vendidos a granel, pastosos y suaves, ofrecidos para el consumo. Se analizaron treinta muestras de helados pastosos vendidos a granel, y treinta muestras de helados suaves, realizándose el conteo de coliformes termotolerantes, Staphylococcus spp. coagulase positiva e investigándose la presencia de Salmonella spp. Se detectaron diez (33%) muestras de helado pastoso y cinco (16%) muestras de helado blando fuera de los límites establecidos por la legislación brasileña. Salmonella spp. se encontró en cuatro muestras (6,7%). Esos resultados destacan la necesidad de una mayor atención a la calidad microbiológica de los helados, con el fin de garantizar la seguridad del consumidor.(AU)

Phenotypical characterization and molecular identification of clinical isolates of Candida tropicalis / Caracterización fenotípica e identificación molecular de aislamientos clínicos de Candida tropicalis

Background. Candida tropicalis is an increasingly important human pathogen which usually affects neutropenic oncology patients with common hematogenous seeding to peripheral organs and high mortality rates. Candida pathogenicity is facilitated by several virulence attributes, including secretion of hydrolytic enzymes; however, little is known regarding the C. tropicalis ability to secrete them and their role in the disease. Aims. To confirm by molecular means the identification of 187 clinical isolates (127 from blood, 52 from urine, and 8 from diverse clinical origins) phenotypically identified as C. tropicalis, and to investigate their in vitro aspartyl proteinase, phospholipase, esterase, hemolysin, DNase and coagulase activities. Methods. The molecular confirmation was performed by ITS sequencing, and the enzymatic determinations were conducted using plate assays with specific substrates, with the exception of coagulase, which was determined by the classical tube test. Results. The majority of the strains exhibited a very strong or strong activity of aspartyl proteinase, phospholipase and esterase. A 4.7% of the bloodstream isolates were hemolysin producers, and all were negative for the coagulase and DNase assays. Conclusions. Very strong activities of aspartyl proteinase, phospholipase and esterase profiles were detected, and a statistical association between phospholipase production and blood and urine isolates was found (AU)

Antecedentes. Candida tropicalis es un patógeno del ser humano cada vez más importante que afecta especialmente a pacientes oncológicos neutropénicos, en los cuales es frecuente la diseminación hematógena del microorganismo a órganos periféricos, lo que conlleva elevadas tasas de mortalidad. La patogenicidad de Candida es facilitada por diversos factores de virulencia, incluyendo la secreción de enzimas hidrolíticas; sin embargo, poco se sabe respecto a la habilidad de C. tropicalis para su secreción, así como el papel que desempeña en la enfermedad. Objetivos. Confirmar por un método molecular la identidad de 187 aislamientos clínicos (127 de sangre, 52 de orina y 8 de orígenes diversos) fenotípicamente identificados como C. tropicalis y estudiar la actividad in vitro de las enzimas proteinasa aspártica, fosfolipasa, esterasa, hemolisina, DNasa y coagulasa. Métodos. La confirmación molecular se llevó a cabo mediante secuenciación del ITS y las determinaciones enzimáticas se llevaron a cabo mediante ensayos en placa con sustratos específicos, a excepción de la coagulasa, que se determinó mediante la clásica prueba en tubo. Resultados. La mayoría de los aislamientos analizados mostraron un perfil de actividad muy fuerte o fuerte de proteinasa aspártica, fosfolipasa y esterasa. El 4,7% de las cepas sanguíneas fue productora de hemolisinas y todas fueron negativas para coagulasa y DNasa. Conclusiones. Se detectaron perfiles con una actividad proteinasa aspártica, fosfolipasa y esterasa muy fuerte entre los aislamientos clínicos analizados, así como también se encontró asociación estadística entre la producción de fosfolipasa y aquellos aislamientos obtenidos de sangre y orina (AU)

Characterization of Staphylococcus spp. strains in milk from buffaloes with mastitis in Brazil: the need to identify to species level to avoid misidentification / Caracterização de cepas de Staphylococcus spp. isoladas de leite de búfalas com mastites em Brasil: a necessidade de se identificar até nível de espécie para evitar erros de identificação

Mastitis is an inflammation of the mammary gland that affects dairy cattle worldwide causing economic losses. Coagulase-negative staphylococci (CNS) are the predominant cause of this type of infection. We have recently showed that coagulase-positive staphylococci could be misidentified. So, the aim of this study was to characterize the Staphylococcus spp. strains initially classified as coagulase-negative Staphylococci, isolated from buffalo with subclinical mastitis. Milk of buffaloes with mastitis in herds was collected and 9 strains were identified as CNS by phenotypic tests. Molecular methodologies latter identified the strains as coagulase-negative Staphylococcus chromogenes (5), coagulase-positive Staphylococcus hyicus (2) and coagulase-positive Staphylococcus aureus (2). Our results strongly support the need to identify the isolates to a species level in order to avoid misidentification and to be aware of the classification using the coagulase test alone.(AU)

A mastite é uma inflamação da glândula mamária que afeta o gado leiteiro em todo o mundo, causando perdas econômicas. Staphylococcus coagulase-negativa (SCN) são a causa predominante desse tipo de infecção. Mostrou-se recentemente que Staphylococcus coagulase-positiva podem ser identificados erroneamente. Assim, o objetivo deste estudo foi caracterizar cepas de Staphylococcus spp. inicialmente classificados como Staphylococcus coagulase-negativa, isolados de búfalas com mastite subclínica. O leite de búfalas com mastite foi coletado, e nove cepas foram identificadas como SCN por testes fenotípicos. Metodologias moleculares identificaram as cepas como Staphylococcus chromogenes coagulase-negativa (5) Staphylococcus hyicus coagulase-positiva (2) e Staphylococcus aureus coagulase-positiva (2). Os resultados reforçam a necessidade de identificar as cepas em termos de espécie, a fim de se evitarem erros de identificação e estar atento à classificação utilizando o teste de coagulase sozinho.(AU)

High prevalence of methicillin-resistant coagulase-negative staphylococci isolated from a university environment in Thailand

The present study was conducted to isolate and characterize the molecular epidemiology of the methicillinresistant staphylococci in the general university environment, where all five locations; the library, restrooms, canteens, computer rooms and outdoor surfaces, are in common use by a large population of students. We used Mannitol Salt Agar (MSA) supplemented with 4 µg/ml of oxacillin to screen the methicillin-resistant staphylococci. The species level was identified by PCR of rdr (Staphylococcus epidermidis), groESL (Staphylococcus haemolyticus) and nuc (Staphylococcus aureus and Staphylococcus warneri) genes and DNA sequencing of tuf and dnaJ genes. The susceptibility patterns of the isolates were determined using the disk diffusion method. Antibiotic and disinfectant resistance genes, together with SCCmec types, were detected by the PCR method. The methicillin resistant-staphylococci were isolated from 41 of 200 samples (20.5%), and all of them were found to be methicillin-resistant coagulase negative staphylococci (MR-CoNS). The library had the highest percentage of contamination, with 43.3% of the samples found to be contaminated. All isolates belonged to 6 different species including S. haemolyticus, S. epidermidis, S. warneri, S. cohnii, S. saprophyticus and S. hominis. The antimicrobial resistance rates were highest against penicillin (100%), then cefoxitin (73.1%), erythromycin (73.1%) and oxacillin (68.3%). Altogether, the isolates were approximately 61.0% multidrug resistant (MDR), with the S. epidermidis isolates being the most multidrug resistant (P < 0.05). The prevalence of the qacA/B gene was detected in 63.4% of the isolates, and SCCmec could be typed in 43.9% (18/41) of the isolates. The type range was: II (n = 1), IVd (n = 1), I (n = 2), V (n = 6), IVa (n = 8) and untypeable (n = 23). This result indicates that these university environments are contaminated with methicillin-resistant coagulase negative staphylococci that carry various SCCmec types and high rate of disinfectant resistance genes (AU)

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Antimicrobial susceptibility profile and research of mec A and erm genes in coagulase-negative staphylococci isolated from platelet concentrates bags

Abstract In recent years, several studies have described the clinical impact of bacterial infection associated with transfusion of platelet concentrates (PCs). Among the blood components, PCs are responsible for the highest rates of bacterial contamination as well as septic transfusion reactions. We assessed antimicrobial susceptibility profile, resistance to methicillin (MRCoNS), and resistance to macrolides, lincosamides and streptogramins of group B (MLSB) of 16 coagulase-negative staphylococci (CoNS) isolates from an investigation in 691 PCs bags. We then compared conventional and automated phenotypic methods, disc diffusion test (DD) and VITEK(r) 2, respectively as well as phenotypic and genotypic methods (Polymerase Chain Reaction - PCR). All CoNS were susceptible to vancomycin. The disc diffusion test characterized 18.75% as MRCoNS and 37.5% with inducible resistance to MLSB (iMLSB), and with VITEK(r) 2, 6.3% and 31.25%, respectively. The mecA gene was detected in 18.75% and the erm gene in 31.25% of the isolates. In this study, we found equal percentage values between presence of the mecA gene by PCR and resistance to methicillin using cefoxitin by DD test, evidence of the erm gene by PCR, and iMLSB resistance by automation (VITEK(r) 2). Moreover, we identified three strains with beta-lactamase overproduction, and the occurrence of a bigger mistake was verified when automation was compared with DD test. And we observed that D-test was the most reliable for the detection of iMLSB resistance in Staphylococcus sp.

Aquisição de imunidade passiva em cabritos alimentados com colostro de cabras com e sem mastite / Immunity of procurement in passive kids fed colostrum from goats with and without mastitis

O objetivo deste trabalho foi avaliar a transferência de imunidade passiva de cabras, que pariram com mastite, para seus respectivos cabritos. Os animais foram distribuídos em dois grupos, a saber: grupo 1 (GI), constituído por cabritos, filhos de cabras sem isolamento microbiológico em ambas as glândulas mamárias, e grupo 2 (GII), composto por cabritos, filhos de cabras com resultado positivo à lactocultura, em pelo menos uma das glândulas mamárias. Foram coletadas amostras de colostro e sangue à parição, bem como às 24 e às 48 horas após o parto/nascimento. O diagnóstico e o monitoramento da mastite nos animais foram realizados por meio do California Mastitis Test (CMT), contagem de células somáticas e isolamento microbiológico. A proteína total foi mensurada pelo método do biureto, e as concentrações de imunoglobulina A (IgA), imunoglobulina G (IgG), transferrina, albumina e haptoglobina por meio da eletrofoerese em gel de poliacrilamida contendo dodecil sulfato de sódio (SDS-PAGE). Os agentes mais isolados na cultura microbiológica foram os Staphylococcus coagulase negativa. Não houve diferença significativa (P<0,05) entre os valores médios de imunoglobulina G (IgG) nos cabritos provenientes de cabras com mastite quando comparados aos recém-nascidos oriundos de cabras livres de infecções intramamárias. Da mesma forma, a atividade de gamaglutamiltransferase (GGT) não mostrou diferença entre os grupos em todos os momentos avaliados. A ingestão de colostro decorrente de cabras com mastite não causou falha na transferência de imunidade passiva nos respectivos conceptos.

The aim of this study was to evaluate the transfer of passive immunity goats kidded with mastitis to their kids. The animals were divided into two groups, namely: Group 1 (GI) containing kids, sons of goats without microbiological isolation in both mammary glands, and Group 2 (GII), composed of kids, sons of goats with positive result to lactoculture in at least one of mammary glands. Colostrum samples and blood were collected after delivery, 24 and 48 hours after delivery / birth. The diagnosis and monitoring of mastitis in animals were performed using the California Mastitis Test (CMT), somatic cell count and microbiological isolation. Total protein was measured by the biuret method, and the concentrations of immunoglobulin A (IgA), immunoglobulin G (IgG), transferrin, albumin and haptoglobin through eletrofoerese polyacrylamide gel containing sodium dodecyl sulfate (SDS-PAGE). The agents most isolated in microbiological culture were coagulase-negative Staphylococcus. There was no significant difference (p <0.05) between the acquisition of immunoglobulin G (IgG) in goats from goats with mastitis compared to infants originating free goat mammary infections. Similarly the gamma glutamyl transferase (GGT) was equal in the comparison between groups in all evaluated moments. The colostrum intake resulting from goats with mastitis caused no failure in the passive transfer of immunity in their fetuses.

Microbiota en teléfonos móviles de médicos oftalmólogos / Microbiota in mobile phones of medical ophthalmologist

Objetivo: Determinar la microbiota en teléfonos móviles utilizados durante la consulta oftalmológica por parte del personal médico, de los pacientes y de los familiares. Métodos: Se analizaron los teléfonos celulares del personal médico y de los pacientes y/o los familiares en el área de consulta de la especialidad. Se realizó una encuesta para evaluar el patrón de uso y la desinfección de los teléfonos móviles. Se tomó una muestra de raspado de los celulares. Las muestras obtenidas fueron inoculadas en medios de cultivo y se incubaron a 37 °C durante 24 h. Se identificó género y especie en los cultivos positivos y se analizaron los resultados obtenidos utilizando estadística descriptiva. Resultados: Se analizaron 71 teléfonos celulares del personal médico y 52 de los pacientes y/o los familiares. Los microorganismos aislados en los teléfonos celulares de los médicos oftalmólogos fueron: estafilococos coagulasa negativa (ECN) 50%, Staphylococcus aureus 32,4%, enterobacterias 4,2%, actinomicetos 4,2 y 9,8% resultaron negativos. Por otro lado, en los teléfonos celulares de los pacientes y los familiares, los microorganismos aislados fueron Staphylococcus aureus 75%, estafilococos coagulasa negativa (ECN) 24% y enterobacterias 1%. Conclusiones: Los resultados obtenidos muestran que los teléfonos celulares, tanto del personal médico como de los pacientes y sus familiares, contienen bacterias consideradas patógenas que podrían establecer una infección. Es relevante establecer una práctica rutinaria de limpieza del teléfono celular y concienciar a la población de los hábitos de higiene, puesto que en ellos queda el cuidado de sus ojos después de la consulta

Objective: To determine the microbiota of mobile phones used during the ophthalmological consultation by medical personnel, patients, and family members. Methods: An analysis was made on the mobile phones of the medical staff and of patients and/or family members in the area of clinical specialty. A survey was conducted to evaluate the pattern of use and disinfection of mobile phones. A smear sample was taken from the mobile phones. The specimens obtained were inoculated in culture media and incubated at 37 °C for 24 hours. Genus and species were identified in the positive cultures and the results obtained were analysed using descriptive statistics. Results: An analysis was made on 71 mobile phones of medical personnel and 52 from patients and/or family members. The microorganisms isolated in the mobile phones of the ophthalmologists were: coagulase-negative staphylococci 50%, Staphylococcus aureus 32.4%, enterobacteria 4.2%, Actinomycetes 4.2%, and 9.8% were negative. On the other hand, in the phones of patients and relatives, the isolated microorganisms were Staphylococcus aureus 75%, coagulase-negative staphylococci 24%, and enterobacteria 1%. Conclusions: The results obtained show that mobile phones, both of the medical staff and of the patients and their relatives, contain bacteria considered pathogenic that could cause an infection. It is important to establish a routine practice of cleaning mobile phones and to make the population aware of hygiene habits, since they are responsible for the care of their eyes after consultation

Perfil bacteriológico de superficies y equipos en un servicio de Fisioterapia / Bacteriological profile of surfaces and equipment in a Physiotherapy Unit

Objetivo: Determinar el perfil bacteriológico en superficies de trabajo, aditamentos y equipos del área de Fisioterapia de una institución prestadora de salud de nivel 1 de complejidad en salud de la ciudad de Popayán, Cauca, Colombia, durante el mes de diciembre del 2015. Métodos: Se realizó un estudio descriptivo a partir de la toma de 13 muestras elegidas al azar entre superficies de trabajo, aditamentos y equipos del área de fisioterapia, el aislamiento se realizó a partir de medios de cultivos no selectivos y la identificación bacteriana por técnicas manuales. Resultados: De 13 muestras obtenidas, el 38,5 por mil fueron negativas y el 61,5 por mil fueron positiva, en las cuales en el 53,9 por mil se encontraron estafilococos coagulasa negativa y en el 7,6 por mil se aísla Micrococcus sp. y Bacillus sp. Conclusiones: La desinfección de las superficies de trabajo, aditamentos y equipos debe realizarse con un agente de mayor eficacia y potencia contra bacterias grampositivas, a fin de reducir contaminación de material inertes y posibles infecciones cruzadas

Objective: To determine the bacteriological profile in work areas, fittings, and equipment of the Physiotherapy area of Popayán, Cauca, Colombia. Methods: A descriptive study was performed based on 13 randomly selected samples from work surfaces, fittings, and equipment in the physiotherapy area. The isolation was performed using non-selective culture media, and bacterial identification was by using manual techniques. Results: Of the 13 samples obtained, 38.5 per-mille were negative and 61.5 per-mille were positive, in which 53.9 per-mille were coagulase negative staphylococcus, and 7.6 per-mille isolated Micrococcus sp. and Bacillus sp. Conclusions: Disinfection of work surfaces, fixtures and equipment should be performed with a more effective and potent agent against gram-positive bacteria, in order to reduce contamination of inert material and possible cross-infection

Influence of carvacrol and thymol on the physiological attributes, enterotoxin production and surface characteristics of Staphylococcus aureus strains isolated from foods

This study evaluated the influence of the phenolic compounds carvacrol (CAR) and thymol (THY) on some physiological characteristics and on the modulation of the secretion of some staphylococcal virulence factors, that is, coagulase and enterotoxin. This study also investigated possible mechanisms for the establishment of the anti-staphylococcal activity of these compounds. Sublethal concentrations (0.3 and 0.15 µL/mL) of CAR and THY inhibited the activity of the enzymes coagulase and lipase and led to a decrease in salt tolerance. At the tested sublethal concentrations, both CAR and THY led to a total suppression of enterotoxin production. The loss of a 260-nm-absorbing material and an efflux of potassium ions occurred immediately after the addition of CAR and THY at 0.6 and 1.2 µL/mL and increased up to 120 min of exposure. Electron microscopy of cells exposed to CAR and THY (0.6 µL/mL) revealed that individual cells appeared to be deformed, with projections of cellular material. The observations of leakage of cellular material and an altered cell surface suggest that gross damage to a cell's cytoplasmic membrane, which results in a disruption in protein secretion, could be responsible for the anti-staphylococcal properties of CAR and THY.

Prevalence, aetiology and antibiotic resistance profiles of coagulase negative staphylococci isolated in a teaching hospital

In this paper we carried out a study about prevalence of the clinically significant coagulase negative staphylococcal (CNS) isolates found in an university hospital. Two hundred four CNS isolates from 191 patients obtained between the period of 1998 to 2002, were studied. About 27 percent (52/191) of the infection cases studied were confirmed as CNS-associated diseases. Blood stream infection (BSI) was the most frequent CNS associated-disease (25 percent; 13/52). The great majority of the BSI was verified in the Neonatal Intensive Care Unit (NICU). The analysis of the 52 patients medical history showed that 85 percent of the BSI was acquired in hospital. Most of the CNS nosocomial infections were associated with the use of indwelling medical devices. The incidence of methicillin-resistance among significant CNS isolates was 38 percent. In this study, a high percentage of exogenous contaminant was verified (60 percent), indicating that contamination of clinical specimens during sample collection is critical.

Characterization of a new coagulase-negative Staphylococcus species (Staphylococcus pettenkoferi) isolated from blood cultures from a hospitalized patient in Porto Alegre, Brazil / Caracterização de uma nova espécie de Staphylococcus coagulase-negativo (Staphylococcus pettenkofer) isolada de hemoculturas de um paciente hospitalizado em Porto Alegre, Brasil

INTRODUCTION: Staphylococcus pettenkoferi was originally isolated and described by Trülzsch et al (2002). In this study, we characterized two isolates of this newly described species. METHODS: Blood cultures were initially processed using the BacT/ALERT® device, and the isolates were initially characterized using the Vitek2 identification system. RESULTS: The initial characterization revealed slow-growing Gram-positive cocci that formed opaque colonies on sheep blood agar. Other phenotypic/genotypic tests were performed. CONCLUSIONS: We would like to emphasize that this new staphylococcus species is phenotypically similar to other CoNS, especially S. auricularis. This could potentially lead to misidentification of these uncommon species.

INTRODUÇÃO: Staphylococcus pettenkoferi foi originalmente isolado e descrito por Trülzsch et al (2002). Neste estudo, caracterizamos dois isolados dessa nova espécie. MÉTODOS: As hemoculturas foram inicialmente processadas usando o instrumento BacT/ALERT® e os isolados foram inicialmente caracterizados pelo sistema de identificação Vitek2. RESULTADOS: A caracterização inicial revelou um coco Gram-positivo de crescimento lento com formação de colônias opacas em agar sangue de carneiro. Outros testes fenotípicos/genotípicos foram realizados. CONCLUSÕES: Gostaríamos de enfatizar que esta nova espécie de Staphylococcus é fenotipicamente similar a outros CoNS, especialmente, S. auricularis, que poderia levar a um erro na identificação dessas espécies incomuns.

Detection of enterotoxin genes of Staphylococcus sp isolated from nasal cavities and hands of food handlers

Food handlers, an important factor in food quality, may contain bacteria that are able to cause foodborne disease. The present study aimed to research coagulase-negative (CNS) and -positive staphylococci (CPS) in 82 food handlers, analyzing nasal and hand swabs, with identification of 62 CNS (75.6 percent) and 20 CPS strains (24.4 percent). Staphylococcal enterotoxins genes were investigated by PCR. In 20 CPS strains, 19 were positive for one or more genes. The percentage of CNS presenting genes for enterotoxins was high (46.8 percent). Despite of the staphylococcal species, the most common gene was sea (35.4 percent), followed by seh and sej (29.2 percent). The detection of new staphylococcal enterotoxins (SEs) genes showed a higher pathogenic potential in this genus. The presence of these gene points out the importance of CNS not only as contaminant bacteria but also as a pathogen.

Perfil microbiológico de peixes e água de cultivo em pesque-pagues situados na região nordeste do estado de São Paulo / Microbiological profile of fish and pond water in fee-fishing located in the northeast region of São Paulo state, Brazil

O objetivo deste trabalho foi determinar o número de coliformes totais, termotolerantes, Staphylococcus coagulase positivo e a presença de bactéria do gênero Salmonella no músculo, no tecido superficial, no trato gastrintestinal de peixes e na água de cultivo de pesque-pagues situados na microbacia do Córrego Rico, SP. Não foi detectado Staphylococcus coagulase positivo em nenhuma amostra de água e peixe. O número mais provável (NMP) de coliformes totais no peixe variou de 2,0 x 10 a >1,1 x 104 NMP.100 mL-1, na água de cultivo variou de 4,2 x 104 a > 2,4 x 105 NMP.100 mL-1. A contagem de coliformes termotolerantes no peixe variou de < 3 a 5,1 x 103 NMP.g-1, na água de cultivo variou de 3,8 x 102 a 2,0 x 104 NMP.100 mL-1. Não houve diferença estatística (P > 0,05) entre as populações de micro-organismos pesquisados na água, pele e trato gastrintestinal, o que reflete uma relação entre a presença dos micro-organismos na água e nesses dois locais analisados. Foi isolada Salmonella sp. em uma amostra de músculo e em duas amostras de trato gastrintestinal. O pescado pode ser veículo de contaminação cruzada, tendo como fonte dos micro-organismos a pele e o trato gastrintestinal para sua própria musculatura. Portanto, o monitoramento da qualidade da água é de suma importância para garantir a produção de peixes com boa qualidade.

The objective of this study was to determine the number of total and thermotolerant coliforms, coagulase-positive Staphylococcus, and the presence of bacteria of the genus Salmonella in the muscle, surface tissue, and gastrointestinal tract of fish, and in the pond water of fee-fishing establishments located in the Córrego Rico microwatershed in the state of São Paulo, Brazil. Coagulase-positive Staphylococcus was not detected in any sample of pond water or fish. The most probable number (MPN) of total coliforms in fish ranged from 2.0 x 10 to > 1.1 x 104 NMP.100 mL-1, while in pond water it ranged from 4.2 x 104 to > 2.4 x 105 NMP.100 mL-1. The count of thermotolerant coliforms in fish ranged from < 3 to 5.1 x 103 NMP.g-1, while in pond water it ranged from 3.8 x 102 to 2.0 x 104 NMP.100 mL-1. There was no statistical difference (P > 0.05) among the studied populations of microorganisms in pond water, surface tissue and gastrointestinal tract, which reflects a relationship between the presence of microorganisms in the water and in the two tissues analyzed. Salmonella sp. was isolated in one sample of muscle and in two samples of gastrointestinal tract. The fish can be a vehicle for cross contamination, having the gastrointestinal tract and the surface tissue as a source of microorganisms for its muscles. Therefore, monitoring the water quality is important to ensure the production of fish with good quality.

Staphylococcus lugdunensis: características clínicas, microbiológicas y sensibilidad antibiótica de 27 casos / Staphylococcus lugdunensis: clinical features, microbiology and antimicrobial susceptibility of 27 cases

Introducción. Staphylococcus lugdunensis es un estafilococo coagulasa negativo (SCN) con características microbiológicas, clínicas, de virulencia y de sensibilidad a antimicrobianos que le hacen ser una especie claramente diferente de otras especies de estafilococos coagulasa negativos. Presentamos las características clínico microbiológicas de S.lugdunensis aislados en 27 enfermos de nuestro hospital. Material y métodos. Se estudiaron los aislamientos de Staphylococcus lugdunensis del año 2004 al 2011. La identificación y antibiograma se realizó por el sistema MicroScan (Siemens). Se revisaron las historias clínicas de los pacientes con aislamientos de S. lugdunensis. Resultados. Se obtuvieron 27 aislamientos de S. lugdunensis procedentes de: abscesos (9), heridas (9), líquido articular (3), sangre (2), líquido peritoneal (1), exudado ótico (1), exudado nasal (1), orina (1). En 20 casos el cultivo fue puro y en 7 mixto. En 8 casos (30%) las muestras procedían de Ginecología, 6 (22%) de Traumatología, 5 (19%) de Cirugía, 3 (11%) de Medicina interna, 2 (7%) de Pediatría, 3 (11%) de otros servicios. En 15 casos (55,5%) existían antecedentes de cirugía o traumatismo reciente. Fueron sensibles a la penicilina 20 (74%) y no hubo ninguna cepa resistente a oxacilina. Conclusiones. S. lugdunensis se ha aislado mayoritariamente en infecciones de piel y tejidos blandos, y en infecciones de heridas post-quirúrgicas. Se ha aislado en cultivo puro en el 74% (20/27) de los casos. Es importante la correcta identificación de S. lugdunensis para evitar que sea descartado como simple estafilococo coagulasa negativo, lo que nos permitirá tener un mejor conocimiento de las infecciones causadas por este microorganismo (AU)

Introduction. Staphylococcus lugdunensis is a coagulase-negative staphylococcus (CNS) with microbiological characteristics, clinical virulence and antimicrobial susceptibility testing, which makes it a distinctly different species from other coagulase-negative staphylococcus species. We present the microbiological and clinical characteristics of 27 S.lugdunensis isolates in patients of our hospital. Material and methods. Staphylococcus lugdunensis isolates collected in our hospital from 2004 to 2011 were studied. Identification and susceptibility testing were performed using the MicroScan (Siemens) system. The clinical records of patients with S.lugdunensis were reviewed. Results. A total of 27 isolates of S.lugdunensis were obtained from the following sources: abscesses (8), wounds (8), joint fluid (3), blood (2), peritoneal fluid (1), ear exudate (1), nasal discharge (1), and urine (1). In 22 cases the culture was pure and in 5 cases mixed. Samples came from Gynaecology in 8 cases (30%), Traumatology in 6 (22%), Surgery in 5 (19%), Internal Medicine in 3 (11%), Paediatrics in 2 (7%), and 3 (11%) from other departments. Just over half (15 cases, 55.5%) had a history of recent surgery or trauma. A total of 20 (74%) were sensitive to penicillin, and none of the isolates was resistant to oxacillin. Conclusions. S. lugdunensis has been isolated mainly in skin and soft tissue infections, as well as in surgical wounds. The microorganism was obtained in pure culture in 20 cases (74%). The proper identification of S.lugdunensis is important in order to avoid being ruled out as simple coagulase-negative staphylococci, and to give us a better understanding of infections caused by this microorganism(AU)

Caracterização molecular dos mecanismos de resistência à linezolida em estafilococos coagulase-negativos e estudo da estabilidade do fenótipo resistente / Linezolid resistance in negative-coagulase staphylococci: characterization and stability of resistant phenotype

Linezolida foi o primeiro fármaco da classe das oxazolidinonas a ser aprovado para o uso clínico. Esta nova oxazolidinona inibe a síntese protéica impedindo a formação do complexo de iniciação formado pelo mRNA, tRNA f-Met e a subunidade 50S do ribossomo bacteriano. Embora a resistência à linezolida possa ser mediada pelo produto do gene cfr ou por mutações nas proteínas ribossômicas L3, L4 e L22, o mecanismo de resistência mais comum envolve mutações no domínio V do gene rRNA 23S. Entre março de 2008 a dezembro de 2011, 38 cepas de estafilococos coagulase-negativos (SCNs) resistentes à linezolida (20 S. epidermidis, 14 S. haemolyticus, 3 S. hominis e 1 S. warneri) isoladas de hemoculturas e pontas de cateter de pacientes internados em dois hospitais terciários do Estado de São Paulo foram incluídas neste estudo para a determinação dos mecanismos de resistência e análise da estabilidade do fenótipo resistente. As cepas de SCNs apresentaram altos níveis de resistência à linezolida (CIMs de 16-128 µg/ml) e foram multi-resistentes, permanecendo sensíveis à vancomicina e teicoplanina. A mutação G2576T foi identificada no domínio V do gene rRNA 23S em todas as cepas de SCNs, exceto em uma cepa de S. haemolyticus. O gene cfr e mutações nas proteínas L4 e L22 não foram detectados. Em relação à proteína L3, todas as cepas de S. epidermidis do hospital A, incluindo a cepa controle sensível à linezolida, apresentaram a substituição Leu101Val, sugerindo que essa mutação seja um marcador clonal dessa população sem envolvimento com a resistência à linezolida. A única cepa proveniente do hospital B (S. epidermidis) foi selvagem para essa proteína ribossômica. Somente uma cepa de S. haemoyticus teve uma mutação no gene rplC, resultando na alteração Val154Leu. Em S. hominis, a mutação Phe147Ile foi identificada em uma cepa, enquanto a associação de Gly139Arg e Met156Thr foi observada nas outras duas cepas dessa espécie. A identificação dessas mutações na proteína L3 de...

Linezolid was the first agent of the oxazolidinone class to be introduced clinically. This oxazolidinone inhibits protein biosynthesis by preventing the formation of the initiation complex that consists of the mRNA, the f-Met tRNA and the 50S subunit of the ribosome. Although linezolid resistance has been mediated by the cfr-encoded product or by ribosomal proteins (L3, L4 and L22), the most common mechanism of resistance involves mutations in the central loop of domain V of the 23S rRNA gene. From March 2008 to December 2011, 38 coagulase-negative staphylococci (CNS) strains (20 S. epidermidis, 14 S. haemolyticus, 3 S. hominis e 1 S. warneri) exhibiting resistance to linezolid were isolated from blood and catheter cultures from patients in two tertiary care hospitals in the State of São Paulo and were included in this study for the ascertainment of the resistance mechanisms to this antimicrobial agent and for the analysis of the stability of this resistance. The strains exhibited high-level resistance to linezolid (MICs 16-128 µg/ml) and all were multidrug resistant, remaining susceptible to vancomycin and teicoplanin. The G2576T mutation in domain V region of 23S rRNA was identified in all isolates, except in a linezolid-resistant S. haemolyticus strain. The cfr gene and mutations in ribosomal proteins L4 and L22 were not detected. Regarding L3 protein analysis, all S. epidermidis strains of hospital A, including the linezolid-susceptible control strain, showed the L3 Leu101Val mutation, suggesting that this alteration is probably not involved in linezolid resistance. The one strain from hospital B (S. epidermidis) was wild-type for this ribosomal protein. Only one S. haemolyticus strain had a mutation in the L3 protein, Val154Leu. Two S. hominis strains showed Gly139Arg/Met156Thr mutations whereas one strain had Phe147Ile in L3 protein. The identification of these mutations in L3 protein of the linezolid-resistant S. haemolyticus and S. hominis strains...

Linezolid, Staphylococcus coagulasa-negativa y el problema de las resistencias / Linezolid, coagulase-negative Staphylococcus and the problem of resistances

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Bacteriemias por Staphylococcus coagulasa negativa: análisis de factores pronóstico e influencia del tratamiento antibiótico / Coagulase-negative Staphylococcus bacteraemia: prognosis factors and influence of antibiotic treatment

Introducción: Staphylococcus coagulasa negativa (SCN) es el microorganismo que se aisla con mayor frecuencia en los hemocultivos (HC) obtenidos en pacientes hospitalizados; su pronóstico se ha asociado a la gravedad clínica del paciente así como a un tratamiento antibiótico empírico inadecuado pero en la actualidad la influencia del tratamiento antibiótico empírico adecuado en la supervivencia de los enfermos no representa un factor pronóstico claramente reconocido. Los objetivos del estudio fueron analizar los factores asociados a una mayor mortalidad en pacientes con bacteriemia por SCN y la influencia del tratamiento antibiótico empírico en su evolución. Pacientes y método: análisis prospectivo (enero a junio de 2010) de los pacientes con HC positivos para SCN en un centro hospitalario universitario; se clasificó la bacteriemia como verdadera atendiendo a criterios de los CDC y se evaluaron los parámetros epidemiológicos, clínicos y microbiológicos relacionados con el fallecimiento del paciente. Resultados: se incluyeron 269 casos en el estudio (97 bacteriemias verdaderas); el 92% de los pacientes evolucionó hacia la curación y el 8% fallecieron (el 1,6% de los fallecimientos se consideró relacionado con la bacteriemia por SCN). Staphylococcus epidermidis fue el SCN identificado con más frecuencia. En el estudio de mortalidad relacionada se incluyeron 93 casos de bacteriemia verdadera. Se asociaron de forma estadísticamente significativa al fallecimiento de los pacientes (estudio bivariante) la gravedad clínica del enfermo (Winston I-III), el ser portador de marcapasos, el desarrollo de sepsis o endocarditis infecciosa y la bacteriemia persistente. El tratamiento empírico adecuado no se asoció a una mayor supervivencia. Conclusiones: el pronóstico de los enfermos con bacteriemia por SCN se asocia a la gravedad y las complicaciones sépticas desarrolladas, siendo mayor en pacientes portadores de marcapasos; en nuestra experiencia el tratamiento empírico inadecuado no se asocia a mortalidad(AU)

Introduction: Coagulase-negative staphylococci (CNS) are the most frequent isolated microorganism in blood cultures; mortality has been associated to severity and to adequacy of empirical treatment but the relevance of the latter is not clearly recognised. The aims of the study were to analyze clinical and microbiological factors related to mortality in patients with CNS bacteraemia and the influence of empirical treatment in prognosis. Patients and methods: a prospective cohort study of patients with CNS bacteraemia was performed (January to June 2010) at a university-affiliated hospital; a determination of clinical significance was made and true bacteraemia was defined according to CDC criteria. We analysed epidemiological, clinical and microbiological variables related to mortality. Results: a total of 269 cases were included (97 were considered true bacteraemia); 92% survived and mortality was 8% (1.6% CNS bacteraemia related mortality). Staphylococcus epidermidis was the most frequent isolated species; 93 patients were included in the related mortality study of patients with true bacteraemia. Factors associated to mortality in the bivariate analysis (p<0.05) were: Winton score I-III, presence of pacemakers, sepsis or infective endocarditis and persistent bacteraemia. Adequate empirical treatment was not associated to survival. Conclusions: severity at onset, the development of septic complications and having a pacemaker are associated to mortality in patients with CNS bacteraemia; in our cohort, inadequate empirical treatment is not related to mortality(AU)

Intramammary infections during the periparturient period in Argentine dairy heifers / Infecciones intramamarias durante el periparto en vaquillonas en Argentina

Prevalence of intramammary infections at prepartum and postpartum in primigravid heifers from five dairy herds located in the central dairy area of Argentina was determined. Mammary secretion samples from 140 heifers (560 mammary quarters) were obtained 14 days prior to the expected calving day and within 7 days after parturition and subjected to bacteriological analysis. No clinical mastitis cases were detected during the study. The number of infected heifers in at least one mammary quarter at pre and postpartum was 87 (62.2%) and 53 (37.8%), respectively. The most prevalent mastitis pathogens at prepartum among samples yielding a positive bacteriological culture were coagulasenegative staphylococci (69.07%), Staphylococcus aureus (12.71%) and Streptococcus uberis (4.42%). A decrease on isolation frequency of coagulase-negative staphylococci (53.41%) and S. uberis (2.27%) was observed at postpartum, while that of S. aureus showed an increase (21.59%). Presence of intramammary infections appeared to be associated with some management conditions. These results highlighted the need to improve diagnosis and control measures in replacement heifers.

Se determinó la prevalencia al preparto y posparto de infecciones intramamarias causadas por organismos patógenos de mastitis en vaquillonas primíparas de cinco establecimientos lecheros ubicados en la cuenca central santafesina. Se tomaron muestras de secreción mamaria de 140 vaquillonas (560 cuartos mamarios) aproximadamente 14 días antes de la fecha probable de parto y dentro de los 7 días posparto, y se procesaron bacteriológicamente. No se detectaron casos de mastitis clínicas durante el estudio. El número de vaquillonas infectadas en al menos un cuarto mamario al preparto y posparto fue de 87 (62,2%) y 53 (37,8%), respectivamente. Los organismos patógenos más prevalentes al preparto entre las muestras con cultivo bacteriológico positivo fueron estafilococos coagulasa negativos (69,07%), Staphylococcus aureus (12,7%) y Streptococcus uberis (4,42%). Al posparto se observó un descenso en la frecuencia de aislamiento de estafilococos coagulasa negativos (53,41%) y S. uberis (2,27%), mientras que la de S. aureus mostró un aumento (21,59%). La presencia de infecciones intramamarias pareció estar asociada con algunas prácticas de manejo. Estos resultados ponen de manifiesto la importancia de hacer extensivo el diagnóstico y control de la enfermedad a las vaquillonas de reemplazo antes de su ingreso al rodeo en ordeño.

Biochemical characteristics of typical and atypical "Staphylococcus aureus" in mastitic milk and environmental samples of brazilian dairy farms

This study results on the biochemical characteristics of 274 "Staphylococcus aureus" strains isolated from mastitic milk, hands of milkmen and milking machines. Features included colony morphology on Baird Parker agar (BPA), catalase, coagulase, and thermonuclease tests. API-Staph (BioMérieux, France) was used as reference identification system. 39.4(per cent) of the colonies on BPA identified as "S. aureus" were atypical. They were more frequent in milk samples (94.4(per cent)). All "S. aureus" strains were coagulase positive but intensify of the test varied according to the source: enviromental strains were 3+ or 4+, while 17.1(per cent) of strains isolated from milk presented only 1+ or 2+ results. The majority of atypical "S. aureus" strains presented a 4+ coagulase result. 2.2(per cent) of "S. aureus" strains were thermonuclease negative. Sixteen thermonuclease positive and coagulase positive strains were identified as "S. hyicus"

Estudio comparativo de las meningitis por Staphylococcus aureus y Staphylococci coagulasa negativos en adultos / Comparative study of meningitis due to staphylococcus aureus and coagulase-negative staphylococci in adults

Introducción. Los Staphylococcus spp. raramente causan meningitis aguda bacteriana (MAB) comunitaria, pero sonuna etiología frecuente de meningitis nosocomial. Objetivo. Descripción y análisis comparativo de MAB por S. aureus y Staphylococci coagulasa negativos (CoN) en adultos. Pacientes y métodos. Revisión del historial de pacientes de edad igual o superior a 14 años diagnosticados de MAB, incluyendo aquéllos con cultivo de líquido cefalorraquídeo (LCR) positivo paraStaphylococcus spp. Resultados. Se incluyen 40 casos (el 28% de las MAB), 12 por S. aureus y 28 por Staphylococci CoN. La incidencia aumentó en el tiempo: el 1,9% en el primer tercio del estudio, el 22,9% en el segundo y el 19,4% en el tercero (p = 0,003). El 67% fue nosocomial y el 95%, postoperatorio. Los dispositivos neuroquirúrgicos (74%) y la neurocirugía reciente(26%) fueron los factores de riesgo, y ambos se asociaron en el 61%. La clínica y alteraciones del LCR fueron similares en ambos grupos, pero en infecciones por S. aureus fue más frecuente la bacteriemia (60% frente a 20%) y el desarrollo de complicaciones(50% frente a 11%). La mortalidad global fue del 15%, superior en infecciones por S. aureus (67% frente a 0%). La mortalidad se asoció también a adquisición espontánea de la infección (25% frente a 0%), desarrollo de complicaciones sépticas (37% frente a 6%), bacteriemia (62% frente a 19%) y no retirar o sustituir el dispositivo neuroquirúrgico (60% frente a 0%).Conclusiones. Los Staphylococcus spp. son una etiología frecuente de MAB en hospitales con neurocirugía, especialmente en el postoperatorio neuroquirúrgico y/o en portadores de dispositivos de drenaje del LCR. El pronóstico en infecciones por Staphylococci CoN es favorable, frente a la elevada mortalidad causada por S. aureus

Introduction. Staphylococcus spp. rarely cause community-acquired acute bacterial meningitis (ABM), but they area frequent cause of nosocomial meningitis. Aims. We describe and conduct a comparative analysis of ABM due to S. aureus and coagulase-negative (CoN) Staphylococci in adults. Patients and methods. We reviewed the medical records of patients aged 14 or over who had been diagnosed with ABM, including those with a positive cerebrospinal fluid (CSF) culture for Staphylococcus spp. Results. Forty cases were included (28% of the ABM), 12 due to S. aureus and 28 due to CoNStaphylococci. Incidence increased over time: 1.9% in the first third of the study, 22.9% in the second and 19.4% in the third (p = 0.003). In 67% of cases it was nosocomial and post-operative in 95%. Neurosurgical devices (74%) and recent neurosurgery (26%) were the risk factors, and both associated 61%. Clinical signs and symptoms and alterations to CSF were similar in both groups, but bacteraemia (60% versus 20%) and the development of complications were more frequent ininfections due to S. aureus. Overall mortality rate was above 15%, and was higher in infections due to S. aureus (67% versus 0%). Mortality was also associated with spontaneous acquisition of the infection (25% versus 0%), the development of septic complications (37% versus 6%), bacteraemia (62% versus 19%) and not withdrawing or changing the neurosurgical device (60% versus 0%). Conclusions. Staphylococcus spp. are a frequent cause of ABM in hospitals with neurosurgery, especially inthe post-operative period following neurosurgical procedures and/or in carriers of CSF drainage devices. Prognosis in infections caused by CoN Staphylococci is favourable, in contrast to the high mortality rate caused by S. aureus