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)

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)

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.

High resistance to Azithromycin- A cause of concern.

Macrolide antibiotics are well established class of antimicrobial agents. Azithromycin due to better tolerance and once a daily dose is commonly prescribed by clinician today. The aim of our study was to analyze the resistance to erythromycin in all gram positive bacteria isolated in department of microbiology Dr. RPGMC Tanda. A result of antibiotic susceptibility testing done by Kir By bauer disc diffusion method were analysed, Of the totral gram positive isolates 54 % were resistant to Azithromycin with maximum resistance seen with Enterococcus species (75 %) . It is recommended that an antibiotic policy should be made and microbiological surveillance cotinued so as to decrease the selective pressure on microorganisms.

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.

Caracterização do cassete cromossômico mec (SCCmec) e análise fenótípica e molecular de resistência a oxacilina em estafilococos coagulase-negativa e Staphylococcus aureus / Characterization of chromosomal cassete mec (SCCmec) analysis and molecular an phenotypic oxacillin in coagulase-negative staphylococci and staphylococcus aureus

Os estafilococos coagulase negativa são patógenos associados a infecções nosocomiais, principalmente bacteremia e sepse. A oxacilina é uma das alternativas de escolha no tratamento das infecções causadas por Staphylococcus spp., contudo, a resistência a esta droga têm se tornado um grande problema nas últimas décadas. O objetivo deste estudo foi caracterizar a sensibilidade à oxacilina em amostras de ECN isoladas de pacientes do Hospital das Clínicas (HC) da Faculdade de Medicina de Botucatu (FMB) por diferentes métodos, determinar a concentração inibitória mínima a antimicrobianos, a triagem de amostras com sensibilidade intermediária à vancomicina e a caracterização do perfil clonal de amostras de MRCoNS isoladas de hemoculturas. Um total de 160 amostras foram analisadas quanto a resistência à oxacilina por meio da técnica de difusão da droga em ágar com disco de oxacilina e cefoxitina, teste de triagem em Ágar Mueller-Hinton com 4 μg/mL de oxacilina e 4% de NaCl, E-test, detecção do gene mecA, e determinação da sensibilidade intermediária a vancomicina em ágar BHI com 4 μg/mL e com 6 μg/mL de vancomicina, além da caracterização do perfil epidemiológico pela tipagem do cassete cromossômico mec (SCCmec) e análise clonal por PFGE de 59 amostras de S. epidermidis e 07 amostras de S. haemolyticus isoladas no período de 2002 a 2009. Um total de 74,4% das amostras foram positivas para o gene mecA. A sensibilidade do método de difusão com disco de oxacilina e da técnica de E-test foi de 99,1% e com especificidade de 90,1%. O disco de cefoxitina apresentou 100% de sensibilidade e a mesma especificidade descrita para o disco de oxacilina. O método de triagem apresentou 94,2% de sensibilidade e a melhor taxa de especificidade com 93,2%. Houve diferença entre as amostras MRCoNS...

Oxacillin is one of the alternatives of choice in the treatment of infections caused by Staphylococcus spp. However, resistance to this drug has become a major problem in recent decades. The aim of this study was to characterize the sensitivity to oxacillin in CNS samples isolated from patients at the School of Medicine University Hospital (HCFMB) - UNESP by different methods, to determine antimicrobial minimum inhibitory concentration, the screening of samples with intermediate sensitivity to vancomycin and characterization of the clonal profile of MRCoNS samples isolated from blood cultures. A total of 160 samples were analyzed for resistance to oxacillin by the drug diffusion technique in agar with oxacillin and cefoxitin, a screening test on Mueller-Hinton agar with 4 μg/mL of oxacillin and 4% NaCl, E-test, mecA gene detection, and determination of intermediate susceptibility to vancomycin in BHI agar with 4 μg/mL and 6 μg/mL of vancomycin, and characterizing the epidemiological profile by typing the cassette chromosome mec (SCCmec) and clonal analysis by PFGE a total of 59 samples of S. epidermidis and 07 samples of S. haemolyticus isolated in the period 2002 to 2009. A total of 74.4% of the samples were positive for mecA. The sensitivity of the oxacillin disk diffusion and E-test technique was 99.1% and specificity 90.1%. The cefoxitin disk showed 100% sensitivity and specificity reported for the same oxacillin disc. The screening method showed 94.2% sensitivity and specificity with the best rate of 93.2%. There were differences between samples when analyzed MSCoNS MRCoNS and the minimum inhibitory concentration (MIC), with higher levels of MIC in the group MRCoNS, and higher rates of multidrug resistance. None of the samples showed resistance or heteroresistant to vancomycin, however, 10 samples grown...

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...

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.

Conventional and molecular characterization of coagulase-negative staphylococcus in hospital isolates.

Background: During the last decade, coagulase-negative staphylococci (CoNS) have emerged as a major cause of nosocomial infections. They constitute a major component of the normal skin and mucosal microflora, and are particularly responsible for catheter- and medical device-related sepsis. They present unique problems in diagnosis and treatment of infections. Purpose: The present study has been designed to evaluate phenotypic and genotypic characteristics of CoNS among nosocomial isolates. Setting and Design: This study was carried out in a tertiary care hospital. Data from 150 samples collected from 73 hospitalized patients and 15 healthy volunteers between October 2003 and May 2005 were analyzed. Patients and Methods: A total of 100 CoNS strains responsible for sepsis or implant-associated infections and 50 saprophytic strains were studied. Invasive CoNS strains were selected on the basis of different colony morphologies, drug resistance patterns, and biofilm formation. The same criteria were used to select saprophytic isolates. Multiplex PCR was used to explore the ica, mecA, and atlE genes, which might contribute to the pathogenicity of CoNS and the formation of biofilms. Results: Most of the invasive strains that formed the biofilm were resistant to multiple antibiotics, with more than 80% resistant to methicillin. ica and mecA genes were detected significantly in pathogenic strains (chi-square test, P<0.0001) whereas atlE was ubiquitously amplified in all the strains. All those strains which had ica and mecA genes were resistant to multiple antibiotics and were positive for biofilm formation. Conclusion: These genetic markers thus appear to discriminate between potential invasive virulent and saprophytic strains of CoNS.

Comparison of four methods for rapid identification of Staphylococcus aureus directly from BACTEC 9240 blood culture system.

Purpose: Differentiation of Staphylococcus aureus (S. aureus) from coagulase-negative staphylococci is very important in blood stream infections. Identification of S. aureus and coagulase-negative staphylococci (CoNS) from blood cultures takes generally 18-24 h after positive signaling on continuously monitored automated blood culture system. In this study, we evaluated the performance of tube coagulase test (TCT), slide agglutination test (Dry Spot Staphytect Plus), conventional polymerase chain reaction (PCR) and LightCycler Staphylococcus MGrade kit directly from blood culture bottles to achieve rapid identification of S. aureus by using the BACTEC 9240 blood culture system. Materials and Methods: A total of 129 BACTEC 9240 bottles growing gram-positive cocci suggesting Staphylococci were tested directly from blood culture broths (BCBs) with TCT, Dry Spot Staphytect Plus, conventional PCR and LightCycler Staphylococcus MGrade kit for rapid identification of S. aureus. Results: The sensitivities of the tests were 99, 68, 99 and 100%, respectively. Conclusion: Our results suggested that 2 h TCT was found to be simple and inexpensive method for the rapid identification of S. aureus directly from positive blood cultures.

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.

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.

Resistência aos antimicrobianos em estafilococos coagulase-negativa isolados de hemocultura / Antimicrobial resistance in coagulase-negative staphylococci isolated of hemocultura

Os estafilococos coagulase-negativa (ECNs) estão entre os microganismos que constituem a microbiota nor al da pele. O seu significado clínico como agente causal de várias infecções em pacientes com materiais implantados e outras infecções hospitalares tem aumentado nas últimas décadas. No presente estudo avaliou-se a resistência de 36 amostras de ECNs, isoladas de hemoculturas de pacientes internados em hospital público da cidade do Rio de Janeiro. As amostras revelaram um perfil de alta resistência às drogas do grupo betalactâmico, e o emprego da cefoxitina, em substituição à oxacilina, para indicação da presença do gene mecA e detecção da resistência a estas drogas, mostrou-se adequado. O perfil de alta resistência observado entre as amostras recomenda maior rigor e critério na seleção de drogas para o tratamento das infecções po ECNs.

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.

Speciation of coagulase negative staphylococcus causing bacterial keratitis.

Thirty-five coagulase negative Staphylococcus (CoNS) cultured from corneal ulcer were speciated and antibiotic sensitivity tested. S epidermidis was the commonest isolate and it was sensitive to ampicillin and vancomycin.

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"

In vitro activities of quinupristin/dalfopristin and eight other antimicrobial agents against 360 clinical isolates from Korea

The emergence of multi-drug resistant gram-positive cocci such as methicillin-resistant (MR) staphylococci, vancomycin-resistant (VR) enterococci, and vancomycin-intermediate resistant S. aureus (VISA) has given new urgency to the development of new antimicrobial agents. One of these is quinupristin/dalfopristin (Q/D). We decided to determine the susceptibility of gram-positive cocci isolated at two university hospitals in Seoul to Q/D and compare the results with eight other antimicrobial agents. We investigated 120 isolates of S. aureus including 49 MRSAs and one VISA, 120 isolates of coagulase negative staphylococci (CNS), 64 E. faecalis and 56 E. faecium, including seven strains of VR E. faecium. Minimum inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs) for several antimicrobials, including vancomycin and Q/D, were determined by broth microdilution. All S. aureus including VISA were susceptible to Q/D. Q/D MIC90 for both methicillin-susceptible S. aureus (MSSA) and MRSA was 0.25 g/mL. 49 (87.5%) of 56 E. faecium including six of seven VR E. faecium were susceptible to Q/D. E. faecalis were not susceptible to Q/D (only 1.5% susceptible), but were inhibited by ampicillin (94% susceptible) or vancomycin (95%). CNS was susceptible to Q/D (96% susceptible) and vancomycin (100% susceptible). One of 38 staphylococci and two of 17 E. faecium were tolerant to Q/D. In conclusion, Q/D showed excellent activity against all species of gram-positive cocci including MRSA, VISA, and VR E. faecium except E. faecalis, and may provide a valuable option for the treatment of infections caused by these emerging nosocomial pathogens of gram-positive cocci.

Predominant phage types of coagulase positive staphylococci in hospital infections.

360 strains of Staphylococcus aureus isolated from various clinical specimens were subjected to bacteriophage typing. 247(68.6%) strains were typable. Among the typable strains 75(20.83%) belonged to phage group I, 45(12.5%) belonged to phage group III, 6(1.67%) belonged to phage group II and 14(3.89%) strains belonged to miscellaneous group. By far, the largest was the mixed group having 107(29.72%) strains. 113 strains (31.4%) were untypable. All the strains were tested for antibiotic sensitivity test. 287 (79.7%) were multiple drug resistant strains.