Identification and antimicrobial suceptibility profile of bacteria causing bovine mastitis from dairy farms in Pelotas, Rio Grande do Sul / Identificação e perfil de suscetibilidade antimicrobiana de bactérias causadoras de mastite bovina em propriedades leiteiras de Pelotas, Rio Grande do Sul

Abstract Mastitis is an inflammatory process of the udder tissue caused mainly by the bacteria Staphylococcus aureus. The indiscriminate use of antibiotics fosters conditions that favor the selection of resistant microorganisms, suppressing at the same time susceptible forms, causing a serious problem in dairy cattle. Given the importance in performing an antibiogram to select the most adequate antimicrobial therapy, the aim of this study was to identify bacteria isolated from cow's milk with mastitis, in dairy farms situated in the city of Pelotas, Rio Grande do Sul, and to determinate the susceptibility profile of these isolates against the antibiotics used to treat this illness. A total of 30 isolates of Staphylococcus spp., were selected from milk samples from the udder quarters with subclinical mastitis whose species were identified through the Vitek system. The susceptibility profile was performed by the disk diffusion assay, against: ampicillin, amoxicillin, bacitracin, cephalexin, ceftiofur, enrofloxacin, gentamicin, neomycin, norfloxacin, penicillin G, tetracycline and trimethoprim. In the antibiogram, 100.0% of the isolates were resistant to trimethoprim and 96.7% to tetracycline and neomycin, three strains of Staphylococcus spp., (10.0%) presented resistance to the 12 antibiotics tested and 24 (80.0%) to at least eight. These results showed the difficulty in treating mastitis, due to the pathogens' resistance.

Resumo A mastite se constitui no processo inflamatório da glândula mamária causada principalmente por bactérias Staphylococcus aureus. O uso indiscriminado dos antibióticos promove condições que favorecem a seleção de micro-organismos resistentes e, ao mesmo tempo, suprime formas suscetíveis, causando um grave problema para a bovinocultura leiteira. Tendo em vista a importância da realização do antibiograma para a seleção da terapia antimicrobiana mais adequada, o objetivo deste estudo foi identificar bactérias isoladas de leite de vaca com mastite, oriundas de propriedades leiteiras localizadas na cidade de Pelotas, RS, bem como determinar o perfil de suscetibilidade desses isolados frente a antibióticos usados para o tratamento desta doença. Foram selecionados 30 isolados de Staphylococcus spp. de amostras de leite provenientes de quartos mamários com mastite subclínica, cujas espécies foram identificadas através do sistema Vitek. O perfil de suscetibilidade foi realizado pela técnica de difusão em disco, frente a: ampicilina, amoxicilina, bacitracina, cefalexina, ceftiofur, enrofloxacina, gentamicina, neomicina, norfloxacina, penicilina G, tetraciclina e trimetoprima. No antibiograma, 100,0% dos isolados foram resistentes a trimetoprima e 96,7% a tetraciclina e a neomicina, três cepas (10,0%) foram resistentes aos 12 antibióticos testados e 24 (80,0%) a pelo menos oito. Esses resultados demonstram a dificuldade encontrada no tratamento da mastite devido à resistência dos agentes patológicos.

Portación de Staphylococcus Aureus en manipuladores de alimentos de servicios gastronómicos de Asunción, Paraguay (2017) / Staphylococcus Aureus in food manipulators of gastronomic services of Asunción, Paraguay (2017)

Introducción: El Staphylococcus Aureus constituye un agente etiológico frecuente de intoxicaciones alimentarias y se encuentra asociado a diversos alimentos. Objetivo: Describir la frecuencia de portación de Staphylococcus Aureus en manipuladores de alimentos de restaurantes de Asunción durante el 2017. Metodología: Se realizó un estudio observacional, prospectivo, descriptivo de corte transversal a manipuladores de alimentos de restaurantes de Asunción durante los meses de mayo a junio del 2017. Se realizaron tres hisopados nasales a cada uno de los 30 manipuladores de alimentos para determinar si la portación era intermitente, permanente u ocasional. Al mismo tiempo fueron realizadas encuestas donde se midieron variables demográficas. Resultados: La prevalencia de portación de Staphylococcus Aureus fue del 33,3 % (10), siendo el 40% (4) esporádica e intermitente y el 20% (2) permanente. En cuanto a las características demográficas de los manipuladores,el 56,66% (17) fueron del sexo masculino, eran solteros 43,33% (13) y el 50% (15) contaban con estudios secundarios completos. El promedio de edad de 29 de los manipuladores fue de 39,5 años con un rango comprendido entre 18 y 85 años. El 23,33% (7) utilizaban guantes a la hora de manipular los alimentos. Además, se pudo observar que el 26,66% (8) utilizaba gorros. Llamó la atención que el 30% (9) de los encuestados no presentaban uñas limpias durante la manipulación. Palabras clave: Staphylococcus Aureus; manipuladores; alimentos.

Introduction: Staphylococcus Aureus is a frequent etiologic agent of food poisoning and is associated with various foods. Objective : To describe the frequency of Staphylococcus Aureus in food handlers of restaurants in Asunción during 2017. Methodology: An observational, prospective, descriptive, cross-sectional study was carried out on food handlers in restaurants in Asuncion during the months of May to June 2017. Three nasal swabs were made to each of the 30 food handlers to determine if the carriage was intermittent, permanent or occasional. At thesame time, surveys were carried out where demographic variables were measured. Results: The prevalence of carrying Staphylococcus Aureus was 33.3% (10), being 40% (4) sporadic and intermittent and 20% (2) permanent. Regarding the demographic characteristics of the manipulators, 56.66% (17) were male, were single 43.33% (13) and 50% (15) had complete secondary education. The average age of 29 of the manipulators was 39.5 years with ranger between 18 and 85 years. 23.33% (7) used gloves when handling food. In addition, it was observed that 26.66% (8) used caps. It was noted that 30% (9) of theres pondents did not have clean nails during handling. Keywords: Staphylococcus Aureus; manipulators; food.

Detección y expresión de superantígenos y de resistencia antimicrobiana en aislamientos obtenidos de mujeres portadoras de Staphylococcus aureus que cuidan y alimentan niños / Detection and expression of superantigens and resistance in isolates from women carriers of Staphylococcus aureus who take care of and feed children

Resumen Introducción. Staphylococcus aureus coloniza mucosas y piel, y causa graves infecciones en el hombre y los animales. Es importante establecer el estatus de portadoras de cepas enterotoxigénicas de este microorganismo en manipuladoras de alimentos, con el fin de prevenir intoxicaciones alimentarias. Objetivo. Establecer las correlaciones entre los genes de enterotoxinas clásicas, el gen tsst-1, la producción de toxinas en cultivo y la resistencia antimicrobiana en aislamientos de S. aureus provenientes de manipuladoras de alimentos que cuidan niños en sus comunidades. Materiales y métodos. Se cultivaron muestras de las fosas nasales y las yemas de los dedos de las manos, y se identificó S. aureus empleando las pruebas de rutina y métodos automatizados. La extracción de ADN se hizo mediante el método de bromuro de cetil-trimetil-amonio (Cetyl-Trimethyl- Ammonium Bromide, CTAB) modificado. Para la detección de superantígenos se emplearon pruebas de reacción en cadena de la polimerasa (PCR) simple y múltiple, y para la de toxinas, estuches comerciales. Resultados. Se encontró que el 22,0 % de los aislamientos correspondía a portadoras de S. aureus: 17,0 % en los aislamientos de fosas nasales; 5,0 % en los de las manos y 6,7 % simultáneamente en los dos sitios. La prevalencia de superantígenos fue de 73,7 %. El genotipo más frecuente fue el sea-tsst-1, con 10,0 %. La resistencia a un solo antibiótico fue de 74,7 % y, a cuatro antibióticos, de 3,2 %; de los aislamientos, el 93,7 % correspondía a cepas productoras de betalactamasas. La detección de genes clásicos y de tsst-1 mediante PCR fue de 48,4 % y la de toxinas en el sobrenadante, de 42,1 %, con una correlación de 95,7 %. Las mayores correlaciones se establecieron entre las toxinas TSST-1 (22/22) y SEA (17/18). La correlación del gen tsst-1 con la proteína y la resistencia fue de 100 %. Todos los aislamientos con el genotipo sea-tsst-1 t fueron resistentes y productores de las toxinas. Conclusión. La tasa de aislamientos de S. aureus toxigénicos y resistentes obtenidos de mujeres que cuidan y preparan alimentos para niños fue de más de 70 %, lo que demostró su gran virulencia y la consecuente necesidad de aplicar estrictamente las normas higiénicas y sanitarias vigentes para evitar el riesgo de intoxicación alimentaria.

Abstract Introduction: Staphylococcus aureus colonizes mucous membranes and skin causing severe infections in humans and animals. It is important to determine carrier status of enterotoxigenic strains of this microorganism in food handlers to prevent food poisoning. Objective: To establish the correlations among classic enterotoxigenic genes, tsst-1 gene, the production of toxins in cultures and antimicrobial resistance in S. aureus isolates from women who handle the food, feed and take care of children in their communities. Materials and methods: Nasal swab and finger samples were cultured and S. aureus was identified using routine methods and automated systems. DNA extraction was done by the CTAB modified method, and superantigen detection by simple and multiplex PCR, while toxins were detected using commercial kits. Results: We found that 22.0% of subjects were S. aureus carriers: 17.0% corresponded to nose samples, 5.0% to hands and 6.7% to both nose and hands. The prevalence of superantigens was 73.7%. The most frequent genotype was sea-tsst-1 with 10%. Resistance to one antibiotic was 74.7%, and to four antibiotics, 3.2%; 93.7% of the isolates were betalactamase-positive. Classical genes and tsst-1 gene were detected by PCR in 48.4% of samples and toxins in supernatant were detected in 42.1% of them with 95.7% of correlation.The highest correlations were established for TSST-1 and SEA with 100% and 94.4%, respectively. The correlation of tsst-1 gene with toxin production and resistance was 100%. All isolates with genotype sea-tsst-1 were toxin-positive and resistant. Conclusion: The rate of toxigenic and resistant S. aureus isolates from women in charge of feeding and taking care of children was higher than 70%, which demonstrates its high virulence. This requires the strict application of hygienic and sanitary regulations in order to avoid the risk of food poisoning.

Relación entre hacinamiento e infecciones comunitarias de piel y partes blandas / Relationship between crowding and community acquired skin and soft tissues infections

La asociación entre diseminación de infecciones comunitarias, pobreza y hacinamiento es bien conocida. En nuestro hospital observamos casos esporádicos de infecciones post-cesárea por cepas de Staphylococcus aureus meticilino resistente de origen comunitario (SAMRCo). En un estudio prospectivo de familias de José C. Paz atendidas en nuestro hospital, investigamos la relación entre antecedentes de infecciones de piel y partes blandas (IPPB) y hacinamiento extremo (hogares con necesidades básicas insatisfechas tipo 3, (NBI 3). Fueron incluidos 264 hogares; 109 (41.3%) tenían historia de IPPB y 59 (22.3%) eran NBI 3. El 61.0% de los hogares NBI 3 y el 35.6% de los hogares no NBI 3 refirieron IPPB (p = 0.00047). Georreferenciamos los domicilios con Google Maps y los ubicamos en un plano de José C. Paz NBI del censo nacional 2010. En barrios con un porcentaje de NBI > 9.7%, el 51.2% de los hogares tuvo antecedentes de IPPB. Cuando este porcentaje era ≤ 9.7% el porcentaje bajó al 31.1% (p = 0.0019). Estos resultados son sugestivos de diseminación comunitaria de estas infecciones asociada a hacinamiento y barrios pobres. Se debe considerar la presencia de SAMRCo en IPPB comunitarias. Por ello, en mujeres procedentes de zonas con alto porcentaje NBI o con antecedentes de IPPB se podría considerar la inclusión de la vancomicina o la clindamicina en la profilaxis de los partos por cesárea.

The association of the spread of community infections with poverty and overcrowding is well known. In our hospital, located in José C. Paz, we assist sporadic cases of post-cesarean infections caused by community acquired methicillin-resistant Staphylococcus aureus (CaMRSA). In a prospective study of families treated at our hospital, we investigated the relationship between a history of skin and soft tissue infections (SSTI) and extreme overcrowding defined as households with unsatisfied basic needs type 3 (NBI 3). A total of 264 households were included in the study; 109 (41.3%) had a history of SSTI and 59 (22.3%) were NBI 3. A total of 61.0% of the NBI 3 households and 35.6% of the non-NBI 3 households reported SSTI (p = 0.00047). Using Google Maps, we georeferenced households and identified them on a NBI map adapted from the 2010 demographic census. In neighborhoods with NBI > 9.7%, 51.2% of the households had a history of SSTI. When NBI was < 9.7%, the percentage fell to 31.1% (p = 0.0019). These results are suggestive of an association of SSTI acquired in the community with overcrowding and poverty. The presence of CaMRSA in community SSTIs should be suspected. Vancomycin or clindamycin prophylaxis could be considered when cesarean deliveries are performed in women from areas with high NBI or with a history of SSTI.

Sphygmomanometers and thermometers as potential fomites of Staphylococcus haemolyticus: biofilm formation in the presence of antibiotics

BACKGROUND The association between Staphylococcus haemolyticus and severe nosocomial infections is increasing. However, the extent to which fomites contribute to the dissemination of this pathogen through patients and hospital wards remains unknown. OBJECTIVES In the present study, sphygmomanometers and thermometers were evaluated as potential fomites of oxacillin-resistant S. haemolyticus (ORSH). The influence of oxacillin and vancomycin on biofilm formation by ORSH strains isolated from fomites was also investigated. METHODS The presence of ORSH on swabs taken from fomite surfaces in a Brazilian hospital was assessed using standard microbiological procedures. Antibiotic susceptibility profiles were determined by the disk diffusion method, and clonal distribution was assessed in pulsed-field gel electrophoresis (PFGE) assays. Minimum inhibitory concentrations (MICs) of oxacillin and vancomycin were evaluated via the broth microdilution method. Polymerase chain reaction (PCR) assays were performed to detect the mecA and icaAD genes. ORSH strains grown in media containing 1/4 MIC of vancomycin or oxacillin were investigated for slime production and biofilm formation on glass, polystyrene and polyurethane catheter surfaces. FINDINGS ORSH strains comprising five distinct PFGE types were isolated from sphygmomanometers (n = 5) and a thermometer (n = 1) used in intensive care units and surgical wards. ORSH strains isolated from fomites showed susceptibility to only linezolid and vancomycin and were characterised as multi-drug resistant (MDR). Slime production, biofilm formation and the survival of sessile bacteria differed and were independent of the presence of the icaAD and mecA genes, PFGE type and subtype. Vancomycin and oxacillin did not inhibit biofilm formation by vancomycin-susceptible ORSH strains on abiotic surfaces, including on the catheter surface. Enhanced biofilm formation was observed in some situations. Moreover, a sub-lethal dose of vancomycin induced biofilm formation by an ORSH strain on polystyrene. MAIN CONCLUSIONS Sphygmomanometers and thermometers are fomites for the transmission of ORSH. A sub-lethal dose of vancomycin may favor biofilm formation by ORSH on fomites and catheter surfaces.

Results of pars plana vitrectomy after complicated phacoemulsification surgery / Resultados pós-vitrectomia via pars plana em pacientes submetidos à cirurgia de facoemulsificação com complicação intraoperatória

Purpose: To identify the causes and outcomes of pars plana vitrectomy (PPV) in patients undergoing phacoemulsification with intraoperative complication and to analyze whether the interval between phacoemulsification and PPV interferes with best-corrected final visual acuity. Methods: This descriptive and retrospective analytical study was conducted in Paraná Eye Hospital in 2013. Data were collected from medical records of 38 patients who underwent complicated phacoemulsification and also required PPV. Results: The most frequent complication as a result of phacoemulsification was posterior capsule rupture, observed in 35 patients (92.10%), followed by capsular bag detachment, in three patients (7.89%). Twenty-eight patients (73.68%) had cortical fragments that were removed during PPV. Twelve patients (31.57%) had their intraocular lens repositioned. PPV was performed on the same day of phacoemulsification in one patient (2.63%), within 1 week in 15 patients (39.47%), between 1 week and 1 month in 13 patients (34.21%), and 1 month after phacoemulsification in 9 patients (23.68%). Conclusion: This study is in agreement with worldwide literature, asserting that major complications of phacoemulsification are posterior capsule rupture and capsular bag detachment, and in addition, there is an improvement in the final visual acuity in almost half the cases, even when there are complications during modern cataract surgery, when complementary appropriate treatment is provided. .

Objetivos: Identificar as causas e os resultados da vitrectomia via pars plana (VPP) em pacientes submetidos à cirurgia de facoemulsificação com complicação intraoperatória, analisando se o tempo cirúrgico entre a facoemulsificação e a VPP interfere na melhor acuidade visual corrigida final. Métodos: Estudo analítico descritivo e retrospectivo realizado no Hospital de Olhos do Paraná em 2013. Os dados foram coletados de prontuários de 38 pacientes que foram submetidos à cirurgia de facoemulsificação complicada e que também precisaram de VPP. Resultados: A complicação intraoperatória mais frequente na cirurgia de facoemulsificação, nos pacientes estudados, foi à ruptura de cápsula posterior, que ocorreu em 35 pacientes (92,10%), seguido de desinserção zonular em 3 pacientes (7,89%). Em 28 pacientes (73,68%) foram encontrados restos corticais, que foram removidos durante a VPP. Em 12 pacientes (31,57%) foi realizado o reposicionamento da lente intraocular. A cirurgia de VPP foi realizada no mesmo dia da facoemulsificação em 1 paciente (2,63%), dentro de 7 dias em 15 pacientes (39,47%), entre 1 semana e 1 mês em 13 pacientes (34,21%) e após 1 mês da facoemulsificação em 9 pacientes (23,68%). Conclusão: O presente estudo encontrou dados semelhantes aos descritos na literatura mundial, que afirmam que as principais complicações da facoemulsificação são a ruptura de cápsula posterior e desinserção zonular; e que a acuidade visual final melhora, em aproximadamente metade dos casos, mesmo após ocorrer complicações na cirurgia de catarata moderna, quando instituído tratamento complementar adequado. .

Impression cytology in the evaluation of ocular surface tumors: review article / Citologia de impressão na avaliação de tumores da superfície ocular: artigo de revisão

Impression cytology (IC) has been widely used as a method for evaluating the ocular surface and superficial cells layers in the diagnosis and follow-up after treatment of several ocular surface tumors of both epithelial and melanocytic origin. Information regarding this can be found in the English-language literature since 1992. Using either cellulose acetate or Biopore membranes for specimen collection, a high correlation has been found between IC and tissue histology. Compared with exfoliative cytology with spatula, IC is less traumatic to the patient’s eye, provides a precise location of the area being studied, and allows accurate observation of the cells the way they exist in vivo. The additional advantage of IC is the preservation of limbal stem cells responsible for continuous corneal epithelium renewal; these can be affected after incisional or excisional biopsy at the corneoscleral limbus, which is the most frequent site of appearance of tumors in the stratified epithelium. Treatment for ocular surface squamous neoplasia has historically included surgery, but nonsurgical interventions have also been adopted. Hence, in certain cases, ophthalmologists may prefer interventions less invasive than surgical biopsy such as of impression cytology for both initial diagnosis and therapeutic monitoring of treatment for ocular surface lesions. Nevertheless, it should be considered that IC may be less helpful if the results conflict with the clinical picture or if the clinical diagnosis is uncertain and results are negative. In such cases, surgical biopsy is required for accurate diagnosis. The purpose of this review is to examine the published literature on the utilization of IC for the diagnosis and management of ocular surface tumors and to discuss the requirement for further investigation on the subject.

A citologia de impressão (CI) tem sido amplamente utilizada como um método de avaliação da superfície ocular e das camadas de células superficiais no diagnóstico e no seguimento após tratamento de vários tumores da superfície ocular de origem epitelial ou melanocítica. As informações podem sem encontradas na literatura em língua inglesa desde 1992. Utilizando-se de membranas de acetato de celulose ou Biopore na coleta dos espécimes, uma alta correlação tem sido encontrada entre a CI e a histologia do tecido. Comparando-se com a citologia esfoliativa, a citologia de impressão é menos traumática para o olho do paciente, fornece uma localização precisa da área estudada e permite ver as células da forma como elas organizam-se in vivo. A vantagem adicional da citologia de impressão é a preservação das células- tronco germinativas responsáveis pela renovação contínua do epitélio da córnea. Elas podem ser afetadas após biópsia cirúrgica na região do limbo que é o sítio mais frequentemente acometido pelos tumores do epitélio estratificado. O tratamento para a neoplasia escamosa da superfície ocular tem sido historicamente a cirurgia, mas intervenções não cirúrgicas também foram adotadas. Por esta razão, em certos casos, oftalmologistas podem recorrer a formas menos invasivas que a biópsia cirúrgica (como a citologia de impressão) tanto para o diagnóstico inicial quanto para o monitoramento terapêutico das lesões da superfície ocular. No entanto, deve-se ter em mente que a citologia de impressão deixa de ser útil quando seu resultado não coincide com o quadro clínico ou quando o diagnóstico clínico é incerto e o resultado da citologia de impressão negativo. Nesses casos, a biópsia cirúrgica deve ser realizada para o diagnóstico. O objetivo desta revisão é examinar a literatura sobre a utilização da citologia de impressão no diagnóstico e tratamento dos tumores da superfície ocular bem como discutir a necessidade de uma investigação mais aprofundada sobre o assunto.

Perfil de sensibilidade de Staphylococcus spp. e Streptococcus spp. isolados de brinquedos de brinquedoteca de um hospital de ensino / Sensitivity profile of Staphylococcus spp. and Streptococcus spp. isolated from toys used in a teaching hospital playroom

Objetivo: Observar a presença de microrganismos dos gêneros Staphylococcus e Streptococcus em brinquedos de uma brinquedoteca de unidade pediátrica hospitalar, bem como analisar o perfil de resistência aos antimicrobianos das cepas isoladas. Métodos: Foram realizadas coletas de 60 brinquedos, utilizando swab umedecido em solução fisiológica, logo após a utilização pelas crianças. As amostras coletadas foram semeadas em meios de cultura para proporcionar o isolamento dos microrganismos e posterior identificação. Foi realizado o antibiograma para todas as bactérias identificadas, com a técnica de difusão em agar. Resultados: O gênero Staphylococcus estava presente em 87% (52/60) dos brinquedos analisados. Foram isoladas 73 cepas, sendo 29% (21/73) de Staphylococcus coagulasepositiva e 71% (52/73) de Staphylococcus coagulase-negativa. Neste estudo, 90,4% das cepas coagulase-negativas apresentaram resistência à penicilina, 65,4% à oxacilina, 28,8% à claritromicina, 61,5% à clindamicina e nenhuma à vancomicina. Das cepas coagulase-positivas, 76,2% apresentaram resistência à penicilina, 23,8% à oxacilina, 23,8% à claritromicina, 47,6% à clindamicina e nenhuma à vancomicina. Não foram detectadas bactérias do gênero Streptoccocus nos brinquedos estudados. Conclusões: Os resultados obtidos demonstraram que os brinquedos podem apresentar contaminação por bactérias potencialmente patogênicas com resistência aos antimicrobianos, representando uma possível fonte de infecção nosocomial para pacientes que normalmente já se encontram debilitados...

Objective: To evaluate the presence of microorganisms of the genus Staphylococcus and Streptococcus on toys in the playroom of a teaching hospital, as well to as analyze the antimicrobial resistance from isolated strains. Methods: Samples were collected from 60 toys, using wet swabs, soon after being used by the children. The samples were inoculated in enriched and selective agar for isolation and later identification of the microorganisms. Antibiogram testing was performed by agar diffusion technique. Results: The genus Staphylococcus was present in 87.0% (52/60) of the toys. Seventy-three strains were isolated, with 29.0% (21/73) coagulase-positive and 71.0% (52/73) coagulasenegative. Among the coagulase-negative strains, 90.4% were resistant to penicillin, 65.4% to oxacillin, 28.8% to clarithromycin, 61.5% to clindamycin, and none to vancomycin. Among the coagulase-positive strains, 76.2% were resistant to penicillin, 23.8% to oxacillin, 23.8% to clarithromycin, 47.6% to clindamycin, and none to vancomycin. The genus Streptococcus was not detected in any of the evaluated toys. Conclusions: Toys can be contaminated with potentially pathogenic bacteria with antimicrobial resistance, representing a possible source of nosocomial infection for patients who are already debilitated...

The occurrence and dissemination of methicillin and vancomycin-resistant Staphylococcus in samples from patients and health professionals of a university hospital in Recife, State of Pernambuco, Brazil

Introduction Methicillin-resistant Staphylococcus aureus (MRSA) strains have been responsible for many nosocomial outbreaks. Within hospitals, colonized employees often act as reservoirs for the spread of this organism. This study collected clinical samples of 91 patients admitted to the intensive care unit (ICU), hemodialysis/nephrology service and surgical clinic, and biological samples from the nasal cavities of 120 professionals working in those environments, of a University Hospital in Recife, in the State of Pernambuco, Brazil. The main objective of this study was to determine the occurrence and dissemination of methicillin- and vancomycin-resistant Staphylococcus spp. Methods The isolates obtained were tested for susceptibility to oxacillin and vancomycin and detection of the mecA gene. In addition, the isolates were evaluated for the presence of clones by ribotyping-polymerase chain reaction (PCR). Results MRSA occurrence, as detected by the presence of the mecA gene, was more prevalent among nursing technicians; 48.1% (13/27) and 40.7% (11/27) of the isolates were from health professionals of the surgical clinic. In patients, the most frequent occurrence of mecA-positive isolates was among the samples from catheter tips (33.3%; 3/9), obtained mostly from the hemodialysis/nephrology service. Eight vancomycin-resistant strains were found among the MRSA isolates through vancomycin screening. Based on the amplification patterns, 17 ribotypes were identified, with some distributed between patients and professionals. Conclusions Despite the great diversity of clones, which makes it difficult to trace the source of the infection, knowledge of the molecular and phenotypic profiles of Staphylococcus samples can contribute towards guiding therapeutic approaches in the treatment and control of nosocomial infections. .

Aislamientos de Staphylococcus aureus sensibles a meticilina relacionados genéticamente con el clon USA300, ¿origen de los aislamientos SARM de genotipo comunitario en Colombia? / Methicillin-sensitive Staphylococcus aureus isolates related to USA300 clone: Origin of community-genotype MRSA in Colombia?

Introducción. USA300 es un linaje genético que se encuentra en aislamientos de Staphylococcus aureus sensibles (SASM) y resistentes a meticilina (SARM). Actualmente, en Colombia las infecciones por SARM en hospitales y en la comunidad son causadas principalmente por un clon con genotipo comunitario (SARM-GC) relacionado genéticamente con el clon USA300. El origen de esta variante es aún desconocido. Objetivo. Identificar y caracterizar aislamientos de S. aureus resistentes y sensibles a meticilina con el fin de aportar información para establecer un posible origen de los aislamientos SARM-GC en Colombia. Materiales y métodos. Se realizó una caracterización de aislamientos SASM relacionados con el clon USA300 detectados a partir de un análisis de 184 aislamientos de S. aureus (90 SARM y 94 SASM) causantes de infecciones. La relación genética de los aislamientos se determinó por electroforesis en gel de campo pulsado (PFGE), tipificación de secuencias multilocus (MLST) y tipificación del gen de la proteína A ( spa ). Resultados. De los 184 aislamientos, 27 (14,7 %) presentaron características moleculares y relación genética con el clon USA300, y de ellos, 18 fueron SARM y nueve fueron SASM. Todos los aislamientos SARM relacionados con este clon albergaban un casete estafilocócico cromosómico mec (SCC mec ) IVc (3.1.2). En ningún aislamiento SASM se detectaron secuencias remanentes de SCC mec o una duplicación del sitio att B que evidenciaran la pérdida del casete. Conclusión. El origen de los aislamientos SARM-GC en Colombia probablemente se encuentre en la diseminación de clones SASM relacionados con el clon USA300 que adquirieron el SCC mec IVc posteriormente.

Introduction: USA300 is a genetic lineage found both in methicillin-resistant (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) isolates. In Colombia, hospital and community MRSA infections are caused by a USA300-related community genotype MRSA (CG-MRSA) clone. The genetic origin of this clone is unknown yet. Objective: To identify and characterize methicillin-resistant (MRSA) and methicillin-sensitive S. aureus (MSSA) isolates in order to improve the information about the origin of the CG-MRSA isolates in Colombia. Materials and methods: USA300-related MSSA isolates were detected and characterized from a study of 184 S. aureus isolates (90 MRSA and 94 MSSA) recovered from infections. The genetic relatedness of the isolates was established by means of pulsed field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and protein A gene typification ( spa typing). Results: Among 184 isolates, 27 (14.7%) showed molecular characteristics and genetic relationship with the USA300 clone, of which 18 were MRSA and nine were MSSA. All USA300-related MRSA harbored Staphylococcal cassette chromosome mec (SCC mec ) IVc (3.1.2). In the MSSA isolates, SCC mec remnants or att B duplicate sites were not detected. Conclusions: In Colombia, the CG-MRSA isolates probably originated in the dissemination of an USA300-related MSSA clone which later acquired SCC mec IVc.

A severe pneumonia due to methicillin resistant Staphylococcus aureus clone USA 300: implications of vertical transmission / Pulmonía severa debido al clon USA 300 del estafilococo dorado resistente a la meticilina: implicaciones de la transmisión vertical

Staphylococcus aureus is an important pathogen in both community and healthcare associated pneumonia. We describe a case of severe pneumonia caused by the methicillin resistant Staphylococcus aureus (MRSA) clone USA 300 in a 44-year old post-partum woman and the subsequent vertical transmission of this virulent organism to her neonate.

El estafilococo dorado (Staphylococcus aureus) es un patógeno importante tanto en la atención a las comunidades como en el cuidado de la salud en relación con la pulmonía. Se describe un caso de pneumonia severa causada por el clon USA 300 del estafilococo dorado resistente a la meticilina (EDRM) en una mujer de 44 anos en periodo de post-parto, y la posterior transmisión vertical de este virulento organismo a su neonato.

Length of exposure to the hospital environment is more important than antibiotic exposure in healthcare associated infections by methicillin-resistant Staphylococcus aureus: a comparative study

OBJECTIVES: Both total antimicrobial use and specific antimicrobials have been implicated as risk factors for healthcare-associated methicillin-resistant Staphylococcus aureus (HCA-MRSA) infection. The aims of this study were: (I) to explore predictors of a new HCA-MRSA infection in comparison with a new healthcare-associated methicillin-sensitive Staphylococcus aureus (HCA-MSSA); (II) to thoroughly assess the role of recent antibiotic use qualitatively and quantitatively. METHODS: The time-period for our study was from October 1997 through September 2001. Through applying strict criteria, we identified two groups of inpatients, one with a new HCA-MRSA infection and one with a new HCA-MSSA infection. We recorded demographic, clinical and antibiotic use-related data up to 30 days before the positive culture date. RESULTS: We identified 127 and 70 patients for each group, respectively. Two logistic regression models were carried out to assess the role of antimicrobial use (qualitatively and quantitatively). In model I, duration of hospital stay, presence of chronic wounds, aminoglycoside and fluoroquinolone use retained statistical significance. In model II, duration of hospital stay and history of intubation during the last month stood out as the only significant predictors of a subsequent HCA-MRSA infection. No significant differences in outcome were noted. CONCLUSIONS: The length of exposure to the hospital environment may be the best predictor of a new HCA-MRSA infection. Use of aminoglycosides and fluoroquinolones may also stand independently along with presence of chronic ulcers and surgical procedures. No independent association between quantitative antibiotic use and subsequent HCA-MRSA infection was documented.

Nasal carriage of Panton Valentine leukocidin-positive methicillin-resistant Staphylococcus aureus in healthy preschool children / Colonización nasal de Staphylococcus aureus Meticilino Resistente portador de la Leucocidina Panton-Valentine en preescolares

Objective Determining the prevalence of nasal carriage of S. aureus, both sensitive to methicillin and resistant to it, in preschool children and evaluating the presence of Panton-Valentine leukocidin genes in the isolates. Methods This was a cross-sectional study in which cultures from anterior nares were obtained from healthy preschool children. Isolates were identified as S. aureus based on morphological and biochemical tests. Antibiotic susceptibility profiles were determined by the disk diffusion method. All the isolates were further analyzed by multiplex PCR to determine the presence of mecA and PVL genes; methicillin-resistant isolates were also SCCmec typed by multiplex PCR. Results Overall S. aureus nasal colonization prevalence was 38.5 % and 4.8 % for methicillin-resistant strains. All the methicillin-resistant isolates carried the genes for PVL; two isolates possessed the SCCmec type IV, two were SCCmec type I and one was SCCmec type II. Conclusion This study revealed high PVL-positive, methicillin-resistant S. aureus colonization prevalence in healthy preschool children from Cartagena, which may play a key role in the epidemiology of community-associated infection by methicillin-resistant S. aureus in healthy children from this particular geographical area.

Objetivo Determinar la prevalencia de colonización nasal de S. aureus, tanto sensible como resistente a meticilina, en niños preescolares y evaluar la presencia de los genes de la leucocidina Panton-Valentine en estos aislamientos. Métodos Estudio de corte transversal en el que se realizaron cultivos de flora nasal de niños preescolares. Los aislamientos fueron identificados como S. aureus con base en su morfología y pruebas bioquímicas. La susceptibilidad a antibióticos se determinó por el método de difusión en disco. Todos los aislamientos fueron analizados por PCR múltiple para determinar la presencia de los genes mecA y PVL, y para la tipificación del casete cromosómico SCCmec de los aislamientos resistentes a meticilina. Resultados La colonización nasal por S. aureus fue 38,5 %, y la de cepas meticilino-resistentes fue 4,8 %. Todos los aislamientos SARM portaban los genes para PVL, dos portaban el elemento SCCmec tipo IV, dos fueron tipo I y uno fue tipo II. Conclusión Encontramos una alta prevalencia de colonización por cepas meticilino-resistentes, PVL-positivos en la población estudiada, lo que podría jugar un papel clave en la epidemiología de las infecciones por S.aureus meticilino-resistente en esta área geográfica.

Caracterización fenotípica y genotípica de la resistencia a meticilina de cepas de Staphylococcus aureus aisladas en Antofagasta / Phenotypic and genotypic characterization of methicillin resistance in strains of Staphylococcus aureus isolated in Antofagasta

Staphylococcus aureus resistant to methicillin (SARM) has been associated with nosocomial infections due to its capacity to develop resistance to multiple antibiotics. There is little information about the SARM which are found in the hospital services of Antofagasta. We studied the phenotypic and genotypic characteristics of methicillin resistance in 38 strains of S. aureus isolated in Antofagasta, identified by coagulase and API Staph tests and by a biochemical test (Ph-system). The susceptibility to antibiotics was studied using the agar dilution technique, identifying SARM strains with discs of oxacillin. Beta-lactamase with nitrocephine, and the gene mecA by means of PCR. Eighty nine percent (34 strains) were SARM with a high resistance to ampicillin, penicillin, erythromycin, claritromycin. gentiamycin, amikacine and ciprofloxacine. All isolates were susceptible to vancomycin and rifampicin. Beta-lactamase was demonstrated in 79% of the SARM strains. Strain typing and resistance patterns revealed a great diversity of PhP-types and antibiotypes in the isolates. Ninety seven percent of the SARM strains had the gene mecA. One PhP-type (C6) was dominant (5 SARM strains) all had the mecA gene, produced beta lactamase and had the same pattern of antibiotic resistance. We conclude that the dominant phenotypes of SARM strains which have the mecA gene and multiple resistance to antibiotics are present in the hospitals of Antofagasta, and sound the alert on the risk of nosocomial transmission of epidemic clones of SARM.

Staphylococcus aureus resistentes a meticilina (SARM) han sido asociados con infecciones nosocomiales por su capacidad para desarrollar resistencia a múltiple antibióticos, existiendo escasa información acerca de SARM que están circulando en los servicios hospitalarios de Antofagasta. Se estudió características fenotIpicas y genotípicas de la resistencia a meticilina en 38 cepas de S. aureus aisladas en Antofagasta, identificadas por tests de coagulasa y API Staph y por tipificación bioquímica (Ph-Sistem). La susceptibilidad a antibióticos se realizo por técnica de dilución en agar, las cepas SARM fueron identificadas con discos de oxacilina, beta-lactamasa por nitrocefina y gen mecA fue detectado pot PCR. El 89% (34 cepas), fueron SARM con una alta resistencia a ampicilina, penicilina, eritromicina, gentamicina, amikacina y ciprofloxacino. Todos los aislados fueron susceptibles a vancomocina y rifampicina. Beta lactamasa fue demostrada en 79% de las cepas SARM. La tipificación y los patrones de resistencia revelaron una alta diversidad de PhP tipos y antibioticos en los aislamientos. El 97% de las cepas SARM albergaban el gen mecA. Un PhP tipo (C6) fue dominante. (5 cepas SARM), todos presentando el gen mecA, produciendo beta lactamasa y mostrando el mismo patrón de resistencia antibiótica. Se concluye que los fenotipos dominantes de cepas SARM que albergan el gen mecA y resistencia múltiples alos antibióticos están circulando en los hospitales de Antofagasta, alertando sobre el riesgo de transmisión intranosocomial de clones epidémicos de SARM.

role of healthcare personnel in the maintenance and spread of methicillin-resistant Staphylococcus aureus

Healthcare workers may acquire methicillin-resistant Staphylococcus aureus [MRSA] from patients, both hospital and home environments, other healthcare workers, family and public acquaintances, and pets. There is a consensus of case reports and series which now strongly support the role for MRSA-carrying healthcare personnel to serve as a reservoir and as a vehicle of spread within healthcare settings. Carriage may occur at a number of body sites and for short, intermediate, and long terms. A number of approaches have been taken to interrupt the linkage of staff-patient spread, but most emphasis has been placed on handwashing and the treatment of staff MRSA carriers. The importance of healthcare workers in transmission has been viewed with varying degrees of interest, and several logistical problems have arisen when healthcare worker screening is brought to the forefront. There is now considerable support for the screening and treatment of healthcare workers, but it is suggested that the intensity of any such approach must consider available resources, the nature of the outbreak, and the strength of epidemiological associations. The task of assessing healthcare personnel carriage in any context should be shaped with due regard to national and international guidelines, should be honed and practiced according to local needs and experience, and must be patient-oriented

Colonização microbiana precoce de pacientes identificados por triagem neonatal para fibrose cística, com ênfase em Staphylococcus aureus / Early microbial colonization of cystic fibrosis patients identified by neonatal screening, with emphasis on Staphylococcus aureus

OBJETIVOS: Avaliar prospectivamente a colonização bacteriana de pacientes com fibrose cística identificados por triagem neonatal. Avaliar a suscetibilidade a antimicrobianos e caracterizar molecularmente as cepas de Staphylococcus aureus isoladas da orofaringe dos pacientes no período do estudo. MÉTODOS: Foram estudados 25 pacientes com fibrose cística, identificados por tripsina imunorreativa e com diagnóstico confirmado por duas ou mais provas de suor, atendidos regularmente no ambulatório de fibrose cística do Hospital de Clínicas da Universidade Federal do Paraná. Foram coletadas amostras de orofaringe com swab e cultivadas por métodos rotineiros; as colônias bacterianas foram identificadas fenotipicamente e testadas quanto à suscetibilidade a antimicrobianos. Os isolados de S. aureus foram submetidos a tipagem molecular por eletroforese em campo pulsado. RESULTADOS: De um total de 234 amostras de orofaringe, S. aureus foi isolado em maior número (76 por cento dos pacientes, 42 por cento das amostras), seguido de Pseudomonas aeruginosa (36 por cento dos pacientes, 16 por cento das amostras) e Haemophilus spp. (76 por cento dos pacientes; 19 por cento das amostras). Dos 19 pacientes colonizados com S. aureus, foram obtidos 73 isolados, 18 oxacilina-resistentes (24,6 por cento), isolados de dois pacientes, com perfis eletroforéticos idênticos ao do clone brasileiro. Os demais isolados oxacilina-sensíveis distribuíram-se entre 18 perfis eletroforéticos distintos. CONCLUSÃO: Observou-se uma maior prevalência de S. aureus, com isolamento mais precoce em relação aos outros patógenos pesquisados. Os isolados multissensíveis distribuíram-se em clones distintos, caracterizando a não transmissibilidade entre as cepas comunitárias. Os S. aureus resistentes a oxacilina isolados apresentaram perfis eletroforéticos idênticos, provavelmente adquiridos no ambiente hospitalar. P. aeruginosa foi pouco freqüente na população estudada.

OBJECTIVES: To assess bacterial colonization prospectively in patients with cystic fibrosis identified by neonatal screening. To assess susceptibility to antimicrobials and to perform the molecular typing of Staphylococcus aureus strains isolated from the oropharynx of patients during the study. METHODS: Twenty-five cystic fibrosis patients receiving regular treatment at the Cystic Fibrosis Outpatient Clinic of Hospital de Clínicas of Universidade Federal do Paraná Brazil, were included in the study. All patients were identified by trypsin-like immunoreactivity and their diagnosis was confirmed by two or more sweat tests. Oropharyngeal swabs were collected and cultured according to routine methods; bacterial colonies were phenotypically identified and their susceptibility to antimicrobials was tested. S. aureus isolates were submitted to molecular typing using pulsed-field gel electrophoresis. RESULTS: Out of 234 oropharyngeal swabs, S. aureus was the most frequently isolated strain (76 percent of patients, 42 percent of swabs), followed by Pseudomonas aeruginosa (36 percent of patients, 16 percent of swabs) and Haemophilus spp. (76 percent of patients; 19 percent of swabs). Seventy-three isolates were obtained from 19 patients colonized with S. aureus, of which 18 were oxacillin-resistant (24.6 percent), isolated from two patients, with the same electrophoretic profiles as that of the Brazilian clone. The remaining oxacillin-sensitive isolates were distributed into 18 electrophoretic profiles. CONCLUSION: There was higher prevalence of S. aureus, with earlier isolation than other pathogens. Multi-sensitive isolates were distributed into different clones, characterizing non-transmissibility among community-acquired strains. The isolated oxacillin-resistant S. aureus showed identical electrophoretic profiles, probably acquired in hospital. P. aeruginosa was not so frequent in the studied population.