ABSTRACT: Listeriosis is an infectious disease caused by bacteria of the genus Listeria, the neurological form being more common in ruminants. There are many reports of listeriosis in small ruminants in the region that includes Brazil, Argentina and Uruguay. However, these diagnoses were mainly based on histological lesions in the central nervous system (CNS) without the isolation and characterization of the involved Listeria strains. The aim of this study was to report sheep and goats listeriosis cases from 2016 to 2021 in northwestern Uruguay. The diagnosis was made according to lesions observed at histopathology, plus Listeria isolation in CNS, identifying it at specie and serotype level. Nine animals (n=9) of three outbreaks and five sporadic cases of listeriosis were studied. Sheep was the species with more cases in relation to goats, and adults were the category most affected. Cases occurred in spring and less frequently in winter. All presented neurological clinical signs and the lesions in the CNS were consistent with suppurative meningoencephalitis and micro-abscesses in the brainstem. In eight of nine CNS samples, Listeria strains were isolated (seven L. monocytogenes and one L. innocua). All the L. monocytogenes isolates carried the inlA gene; serotyping showed that four strains belonged to serotype 1/2b, two isolates belonged to serotype 4b, and one to serotype 1/2a. Considering that listeriosis is a common disease in this region and the fact that isolates are scarcely recovered from small ruminants, it would be important to emphasize the need for Listeria isolation to better characterize the strains that affect animals. Not only to improve knowledge about the epidemiology of disease but also with the objective of developing serotype specific vaccines for animal use.

RESUMO: Listeriose uma doença bacteriana causada pelo gênero Listeria, a forma nervosa é a mais comum em ruminantes. No Brasil, Argentina e Uruguai há vários relatos de listeriose em pequenos ruminantes com diagnóstico baseado na histopatologia do sistema nervoso central (SNC), sem o isolamento e a caracterização do agente. O objetivo deste trabalho foi relatar uma série de casos diagnosticados em ovinos e caprinos no período 2016-2021 no noroeste do Uruguai. O diagnóstico foi feito basado nas lesões observadas na histopatologia, e caracterização das cepas de Listeria recuperadas do SNC quanto à espécie e sorotipo. Nove animais (n=9) do três surtos e cinco casos isolados de listeriose foram estudados. Os ovinos foram a espécie com o maior número de casos em relação aos caprinos, sendo os animais adultos a categoria mais afetada em ambas espécies. A doença ocorreu principalmente na primavera com alguns casos observados no inverno. Todos os casos apresentavam sinais clínicos nervosos e as lesões no SNC caracterizavam-se por meningoencefalite supurativa com presença de microabscessos no tronco encefálico. Em oito de nove amostras do SNC foram isoladas cepas de Listeria (sete L. monocytogenes e uma L. innocua). Todos os isolados de L. monocytogenes continham o gene inlA; a sorotipagem apresentou quatro cepas do serotipo 1/2b, duas cepas serotipo 4b e uma cepa 1/2a. Levando em consideração que nesta região a listeriose é uma doença frequente e que existem poucos isolados recuperados de casos clínicos em pequeño ruminantes, torna-se relevante o isolamento deste agente para caracterização das cepas que afetam os animais. Não só para melhorar o conhecimento sobre a epidemiologia da doença, mas também com o objetivo de desenvolver vacinas sorotipo-especificas para uso animal.

Abstract The aim of this study was to characterize phenotypically and genotypically 27 mecApositive Staphylococcus aureus strains with oxacillin MICs of ≤2 g/ml by Vitek 2, isolated indifferent regions of Uruguay. Susceptibility to oxacillin and cefoxitin was studied by gradient dif-fusion, disk diffusion to cefoxitin, and Phoenix and MicroScan systems. PBP2a was determined.SCCmec typing was performed and the isolates were compared by PFGE. Twenty-six isolateswere susceptible to oxacillin; one strain was susceptible to cefoxitin by disk diffusion and 3strains by gradient diffusion. Phoenix and MicroScan panels detected methicillin resistance in25 and 27 strains, respectively. Twenty-six strains tested positive for PBP2a. Twenty-six strainscarried SCCmec V and 24 belonged to pulsotype A. One strain carried SCCmec IV and did notbelong to pulsotype A. Cefoxitin disk diffusion test and PBP2a detection correctly identified 26of these 27 strains as MRSA. PFGE results suggest the dissemination of a cluster of MRSA carryingSCCmec V.

Resumen El objetivo de este estudio fue caracterizar fenotípicamente y genotípicamente 27 cepas de Staphylococcus aureus positivas para mecA y con CIM de oxacilina <2 pg/ml según Vitek 2, obtenidas en diferentes regiones del país. La sensibilidad frente a la oxacilina y la cefoxitina se estudió por difusión en gradiente, por disco-difusión (cefoxitina) y por los sistemas Phoenix y MicroScan. Se analizó la portación de PBP2a, se realizó la tipificación de SCCmec y las cepas se compararon mediante PFGE. Resultaron sensibles a oxacilina por difusión en gradiente 26 cepas; una fue sensible a cefoxitina por disco-difusión y 3 lo fueron por difusión en gradiente. Los sistemas Phoenix y MicroScan detectaron resistencia a meticilina en 25 y 27 cepas, respectivamente. Asimismo, 26 cepas portaban PBP2a y 26 cepas mostraron presencia de SCCmec V, 24 correspondieron al pulsotipo A. Una portaba SCCmec IV y no perteneció al pulsotipo A. La prueba de disco-difusión con cefoxitina y la detección de PBP2a identificaron 26 de 27 cepas como MRSA. La PFGE sugiere la diseminación de un grupo MRSA con SCCmec V. © 2022 Asociación Argentina de Microbiología. Publicado por Elsevier Espana, S.L.U. Este es un artículo Open Access bajo la licencia CC BY-NC-ND (https://creativecommons.org/licenses/by-nc-nd/4.0/).

Resumen Las infecciones intrahospitalarias (IIH) son causa de elevada morbimortalidad y representan un problema sanitario importante. El personal de salud es reservorio y potencial transmisor de los agentes etiológicos de las mismas. S. aureus es uno de los microorganismos implicados, por lo tanto es importante conocer la frecuencia de portación en el personal de salud y establecer el perfil de susceptibilidad antimicrobiana para contribuir con la elaboración de medidas de prevención incluyendo actividades educativas. Objetivo: Conocer la frecuencia de portación de S. aureus, distribución y antibiotipos de las cepas presentes en el personal sanitario del Hospital Pediátrico de Referencia (HPR). Materiales y métodos: Se realizó un estudio descriptivo durante el periodo julio-setiembre del año 2018. Se incluyeron muestras de hisopados nasales de trabajadores de la salud de distintas áreas de internación que consintieron participar en el estudio. Se excluyeron aquellos que recibieron antibióticos dentro de los 3 meses previos al estudio. Las muestras fueron sembradas en agar sangre ovina al 5% (ASO) y se incubaron a 35-37ºC en aerobiosis por 24-48 horas. La identificación de las colonias sospechosas de Staphylococcus aureus por métodos convencionales y MALDI-TOF. El patrón de resistencia antimicrobiana de S. aureus se detectó por disco-difusión. En los cultivos resistentes a meticilina (SAMR) se determinó la presencia del gen mecA y se realizó la tipificación del SCCmec por pruebas de reacción en cadena de polimerasa. Resultados: Se obtuvieron 225 hisopados a partir de 225 trabajadores, presentaron desarrollo 212. En 49 se recuperaron cultivos de S. aureus. Correspondieron a SAMR 11 de las 49 cepas, todas portaban el gen mecA. Hubo predominio en el personal de enfermería (7/11), en los servicios de hemato-oncología (3/11) y cuidados intensivos neonatales (4/11). Asociaron resistencia a macrólidos y clindamicina 8 de 11 aislamientos SAMR, a gentamicina 2 y a mupirocina uno. El SCCmec más frecuentemente identificado fue el tipo IV (7/11). Conclusiones: Los resultados muestran la presencia de cepas SAMR entre el personal de salud del CHPR y aportan información complementaria para efectuar prevención y control de las IIH, actuando sobre todo en el personal de salud encargado de la atención de pacientes susceptibles.

Abstract Hospital-acquired infections (IIH) are a cause of high morbidity and mortality and represent a major health problem. Health personnel are reservoirs and potential transmitters of their etiological agents. S. aureus is one of the microorganisms involved, therefore it is important to know the frequency of carriage in health personnel and establish the antimicrobial susceptibility profile to contribute to the development of prevention measures, including educational activities. Objective: To know the frequency of carriage of S. aureus, distribution and antibiotypes of the strains present in the health personnel of the Reference Pediatric Hospital (HPR). Materials and methods: A descriptive study was carried out during the period July-September 2018. Nasal swab samples from health workers from different hospitalization areas who agreed to participate in the study were included. Those who received antibiotics within 3 months prior to the study were excluded. The samples were seeded in 5% sheep blood agar (ASO) and incubated at 35-37ºC in aerobiosis for 24-48 hours. Identification of suspicious Staphylococcus aureus colonies by conventional methods and MALDI-TOF. The antimicrobial resistance pattern of S. aureus was detected by disc diffusion. In methicillin-resistant cultures (MRSA), the presence of the mecA gene was determined and SCCmec was typified by polymerase chain reaction tests. Results: 225 swabs were obtained from 225 workers, 212 showed development. S. aureus cultures were recovered from 49. 11 of the 49 strains corresponded to MRSA, all of them carried the mecA gene. There was a predominance in the nursing staff (7/11), in the hematology-oncology services (3/11) and neonatal intensive care (4/11). They associated resistance to macrolides and clindamycin in 8 of 11 MRSA isolates, 2 to gentamicin, and 1 to mupirocin. The most frequently identified SCCmec was type IV (7/11). Conclusions: The results show the presence of MRSA strains among the health personnel of the CHPR and provide complementary information to carry out prevention and control of IIH, acting especially on the health personnel in charge of the care of susceptible patients.

Resumo As infecções hospitalares (HII) são causa de alta morbidade e mortalidade e representam um importante problema de saúde. Os profissionais de saúde são reservatórios e potenciais transmissores de seus agentes etiológicos. O S. aureus é um dos micro-organismos envolvidos, por isso é importante conhecer a frequência de portadores em profissionais de saúde e estabelecer o perfil de suscetibilidade antimicrobiana para contribuir no desenvolvimento de medidas de prevenção incluindo atividades educativas. Objetivo: Conhecer a frequência de portadores de S. aureus, distribuição e antibiótipos das cepas presentes no pessoal de saúde do Hospital Pediátrico de Referência (HPR). Materiais e métodos: Foi realizado um estudo descritivo durante o período de julho a setembro de 2018. Foram incluídas amostras de swab nasal de profissionais de saúde de diferentes áreas de internação que concordaram em participar do estudo. Aqueles que receberam antibióticos nos 3 meses anteriores ao estudo foram excluídos. As amostras foram semeadas em 5% de ágar sangue de carneiro (ASO) e incubadas a 35-37ºC em aerobiose por 24-48 horas. Identificação de colônias suspeitas de Staphylococcus aureus por métodos convencionais e MALDI-TOF. O padrão de resistência antimicrobiana de S. aureus foi detectado por difusão em disco. Em culturas resistentes à meticilina (MRSA), a presença do gene mecA foi determinada e SCCmec foi tipificado por testes de reação em cadeia da polimerase. Resultados: 225 swabs foram obtidos de 225 trabalhadores, 212 apresentaram desenvolvimento. Culturas de S. aureus foram recuperadas de 49. 11 das 49 cepas correspondiam a MRSA, todas carregavam o gene mecA. Houve predominância na equipe de enfermagem (7/11), nos serviços de hematologia-oncologia (3/11) e de terapia intensiva neonatal (4/11). Eles associaram resistência a macrolídeos e clindamicina em 8 de 11 isolados de MRSA, 2 à gentamicina e 1 à mupirocina. O SCCmec mais frequentemente identificado foi o tipo IV (7/11). Conclusões: Os resultados mostram a presença de cepas de MRSA entre os profissionais de saúde do CHPR e fornecem informações complementares para realizar a prevenção e controle da HII, atuando principalmente sobre os profissionais de saúde responsáveis pelo atendimento de pacientes suscetíveis.

Abstract This study was undertaken to investígate the resistance phenotypes to macrolide-lincosamide-streptogramin B (MLSb) antibiotics and their associated genotypes in isolates of Staphylococcus aureus. We analyzed one hundred, consecutive, non-duplicate isolates (methicillin-susceptible MSSA, n = 53 and methicillin-resistant MRSA, n =47) obtained from var-ious clinical samples between July 2012 to December 2013. The resistance profile to MLSb antibiotics was determined by phenotypic methods and the resistance genes were detected by PCR assays. All of the isolates were subjected to pulsed-field gel electrophoresis (SmaI-PFGE). The overall prevalence of resistance to MLSb antibiotics was 38% and the resistance phenotype distribution was as follows: cMLSb, 22%; iMLSB, 10%; MSb, 5% and L, 1%. We detected ermA, ermC, ermB and mrsA/B genes in these resistant isolates. The single ermA gene was commonly observed mainly in those with a cMLSb R phenotype, whereas the combination ermA and ermC was more commonly observed in isolates with inducible expression. The patterns of SmaI-PFGE suggest a great genetic diversity in both MRSA and MSSA resistant to MLSb antibiotics. The results demonstrate the local presence of S. aureus resistant to MLSb antibiotics and its most frequently described responsible genes. Some of these isolates, especially those with the iMLSB phenotype, may be associated with therapeutic failure. Therefore, efforts should be directed to the correct detection of all MLSb resistant isolates using appropriate laboratory tests. PFGE results reveal a wide spread of resistance genes rather than the circulation of S. aureus clones resistant to MLSb antibiotics.

Resumen Los objetivos de este estudio fueron investigar en Staphylococcus aureus la presencia de fenotipos resistentes a los antibióticos macrólidos, lincosamidas y estreptograminas tipoB (MLSb) y conocer sus genotipos responsables. Analizamos 100 aislamientos consecutivos, no duplicados (53 sensibles a meticilina [MSSA] y 47 resistentes a meticilina [MRSA]), obtenidos entre 2012 y 2013 a partir de diferentes muestras clínicas. El perfil de resistencia a los antibióticos MLSb fue determinado por métodos fenotípicos y los genes de resistencia se detectaron por PCR. Todos los aislamientos fueron comparados por SmaI-PFGE. La prevalencia global de resistencia a los antibióticos MLSB fue del 38% y la distribución de los fenotipos de resistencia fue la siguiente: cMLSB, 22%; iMLSB, 10%; MSB, 5%; L, 1%. Se detectaron los genes ermA, ermC y mrsA/B en los aislamientos resistentes. El gen ermA se observó, sobre todo, en aislamientos con fenotipo resistente constitutivo R (cMLSB), mientras que la combinación ermA y ermC se detectó principalmente en aislamientos con resistencia inducible (iMLSB). Los patrones de Smal-PFGE sugieren una gran diversidad genética en los aislamientos resistentes a los antibióticos MLSb, tanto MRSA como MSSA. Los resultados demuestran la presencia local de S. aureus resistentes a los antibióticos MLSB y de sus genes responsables más frecuentemente descritos. Estos cultivos, especialmente aquellos con fenotipo resistente iMLSB, pueden asociarse con fallas terapéuticas. Por lo tanto, los esfuerzos deben dirigirse a la correcta detección de todos los cultivos resistentes a MLSB utilizando pruebas de laboratorio adecuadas. Los resultados de Smal-PFGE sugieren una amplia diseminación de genes de resistencia, más que la circulación de clones resistentes a los antibióticos MLSB.

ABSTRACT The aim of this study was to describe the microbiological characteristics and profile of genes encoding enterotoxins in 95 Staphylococcus aureus isolates obtained between April 2011 and December 2014 from foodstuffs, persons and surfaces of retail food stores. After microbiological identification and antimicrobial susceptibility testing, polymerase chain reactions (PCR) were performed, targeting sea, seb, sec, sed and see genes that code for classical enterotoxins (ET) A-E, and three additional genes: seg , seh and sei , coding for so-called “new enterotoxins” G, H and I. The isolates were characterized by Pulsed Field Gel Electrophoresis (PFGE), and five selected isolates were further analyzed through Multi Locus Sequence Typing (MLST). It is noteworthy that 54.7% of the examined isolates harbored one or more of the investigated ET gene types. Most positive isolates carried more than one ET gene up to five types; seg was the most frequent ET gene, followed by sei. Five enterotoxin-coding isolates also coded for some antimicrobial resistance genes. Two of them, and four additional non-enterotoxic isolates carried erm genes expressing inducible clindamycin resistance. PFGE-types were numerous and diverse, even among enterotoxin-coding strains, because most isolates did not belong to known foodborne outbreaks and the sampling period was long. MLST profiles were also varied, and a new ST 3840 was described within this species. ST 88 and ST 72 enterotoxin-coding isolates have been identified in other regions in association with foodborne outbreaks. This manuscript reports the first systematic investigation of enterotoxin genes in S. aureus isolates obtained from foodstuffs and infected people in Uruguay.

Abstract Leptospirosis is important in Uruguay due to the economic loss caused by the diseases of production animals, mainly bovines, and also due to frequent human infection. We decided to study anti-Leptospira antibodies in the sera of dairy workers, rice laborers, veterinarians, suburban slum dwellers and garbage recyclers. Our aims were to estimate the seroprevalence of infection by Leptospira spp. in these people at risk, the relative importance of the known risk factors associated with infection, and the impact of human infections in each setting. Groups at risk were identified and 35 visits to their locations were made, conducting field surveys and exchange talks for information and education. Simple epidemiological questionnaires were administered and sera samples were taken from 308 persons. The microagglutination Technique (MAT) and the IgM Indirect Immunofluorescence (IIF) assay were employed to detect antibodies. Environmental water samples, canine and equine sera were also examined. More than 45% of human sera were reactive and the studied groups were confirmed to be widely exposed to infection. Female sera were frequently reactive, though most illnesses occur in men, and the most severe cases in elderly males; the emergence and evolution of the disease may strongly depend on the host condition and functions. Animal contact and unsafe water usage were the main identified risk factors to be considered in prevention. Fifty per cent of the studied horses showed a positive MAT reaction. The underdiagnosis of the illness and its long-term symptoms require further study, as well as greater health and social attention efforts.

Resumen La leptospirosis es importante en Uruguay por las pérdidas económicas que ocasionan las enfermedades en los animales de producción, principalmente en los bovinos, y a causa de la frecuente infección humana. Decidimos estudiar anticuerpos anti -Leptospira en trabajadores de tambo y de plantaciones de arroz, así como en veterinarios, habitantes de asentamientos y recicladores de residuos. Buscamos evaluar la importancia de distintos factores de riesgo conocidos asociados con la infección por Leptospira spp., y estimar la frecuencia y el impacto de las infecciones humanas en la población expuesta. Se efectuaron 35 visitas a colectivos de riesgo, realizando relevamientos de campo e intercambios educativos, llenando formularios epidemiológicos sencillos y tomando muestras de sangre a 308 personas. Se investigaron anticuerpos séricos con técnicas de microaglutinación (MAT) e inmunofluorescencia indirecta para IgM. Se examinaron también, muestras de agua ambiental, y sueros caninos y equinos. Más del 45% de los sueros humanos fueron reactivos y se confirmó que los grupos estudiados estaban ampliamente expuestos a la infección. Los sueros de mujeres fueron frecuentemente reactivos, aunque la mayoría de las enfermedades ocurren en varones, y los casos más graves en varones añosos. La emergencia y la evolución de la afección pueden depender fuertemente del estado del huésped y su respuesta. El contacto con animales y el uso de agua insegura fueron los principales factores de riesgo identificados para considerar en la prevención. El 50% de los sueros equinos fueron reactivos por MAT. Es necesario incrementar los esfuerzos de atención social y sanitaria en estos sectores, superar el subdiagnóstico y estudiar la evolución y la sintomatología a largo plazo de los pacientes.

RESUMO: O objetivo deste trabalho foi detectar a presença de bactérias do gênero Listeria e particularmente Listeria monocytogenes, em bovinos leiteiros no ambiente de Paysandú, Uruguai. Foi realizada uma amostragem em duas etapas, dez estabelecimentos foram selecionados por probabilidade proporcional ao tamanho. Foi realizada uma única visita a cada propriedade. Foram coletadas amostras para cultura bacteriológica de matéria fecal bovina, além de alimentos, leite do tanque de resfriamento, água e solo na entrada e saída da sala de ordenha. Com os isolados de L. monocytogenes foi realizado PCR para a confirmação da espécie e determinação do perfil do serotipo. AscI-elctroforese em gel de campo pulsado foi realizado para compará-los geneticamente. Listeria spp. foram isoladas de oito de dez estabelecimentos, enquanto L. monocytogenes foram detectadas em três deles. A distribuição dos serotipos nos isolados de L. monocytogenes recuperados foi: 1/2a três isolados, 4b um isolado. Foram detectadas vacas leiteras clinicamente sadias que excretaram L. monocytogenes ou L. innocua nas fezes. Dos alimentos do gado houve só um isolamento de L. monocytogenes 1/2a em uma pastagem. Esta estirpe foi idêntica no PFGE a um isolado 1/2a obtido de uma “piscina” de fezes de vacas leiteiras do mesmo estabelecimento. Não houve isolamento de Listeria no leite do tanque de resfriamento ou na água de nenhum dos estabelecimentos. Listeria innocua foi detectada em 13 alimentos para o gado e sete amostras de solo na entrada e saída da sala de ordenha. Este parece ser o primeiro estudo local que confirma a presença de Listeria spp. incluindo L. monocytogenes em vacas leiteiras sadias e no meio ambiente de propriedades leiteiras com base alimentícia na pastagem. Esses resultados sugerem o potencial de vacas sadias e seus subprodutos como possível fonte desses agentes para humanos e/ou outros animais. São necessários estudos mais detalhados que incluem a comparação genética de isolados humanos e de animais para estabelecer claramente seu relacionamento epidemiológico.

ABSTRACT: The aim of the study was to detect Listeria spp., particularly Listeria monocytogenes, in cattle and environment of pasture based dairy farms in Paysandú, Uruguay. A two-stage sampling was conducted, 10 farms were selected by probability proportional to size. A single visit was made to each farm. Samples from bovine faeces, feedstuffs, bulk tank milk, drinking water and soil from the entry and exit pens of the milking parlour were collected for bacteriological studies. PCR assays were used to confirm species and determine the serotype profile of L. monocytogenes isolates. AscI-pulsed-field gel electrophoresis was done to genetically compare them. Listeria spp. were isolated from eight of ten dairy farms, whereas L. monocytogenes in three of them. Serotype distribution in L. monocytogenes was as follows: 1/2a, three isolates; 4b, one isolate. L. monocytogenes or L. innocua excreted from clinically healthy milking cows was detected via faeces. In feedstuffs, only one L. monocytogenes 1/2a isolate from a pasture was obtained. The strain was identical by PFGE to an isolate 1/2a obtained from a pool of milking cow feces that grazed on this farm. No isolation of Listeria spp. was retrieved from the bulk tank milk or drinking water from any of the farms. Listeria innocua was detected in 13 feedstuffs and seven samples of soil from the entry and exit pens of the milking parlour. This is a first local study that confirms the presence of Listeria spp. including L. monocytogenes in healthy cattle and environment of pasture-based dairy farms. These results suggest the potential role that healthy cattle and their sub-products would play as a source of these agents for humans and/or others animals. More detailed studies that include genetic comparison of human and animal isolates are required in order to clearly establish the epidemiological relationship.

ABSTRACT The aim of this work was to study the prevalence of Listeria monocytogenes in foods obtained in retail shops and food industries located in Montevideo-Uruguay, and to identify the serogroups of the obtained isolates. Three-thousand one-hundred and seventy-five food samples (frozen, deli meats, ready-to-eat and cheese) were analyzed. The obtained isolates were serogrouped by multiplex PCR and serotyped by conventional procedure. Genetic comparisons were performed using pulsed-field gel electrophoresis on a sub-set of isolates belonging to the same serotype successively recovered from the same establishment. L. monocytogenes was isolated from 11.2% of samples. The highest prevalence was observed in frozen foods (38%), followed by cheese (10%). 1/2b and 4b were the most frequently identified serotypes. In six of 236 analyzed establishments we successively recovered L. monocytogenes isolates belonging to the same serotype. Most of them corresponded to serotype 1/2b. Pulsed-field gel electrophoresis profiles suggest that at least 33% of L. monocytogenes 1/2b isolates are genetically related and that may remain viable for prolonged periods. The observed prevalence of L. monocytogenes was lower than reported in neighboring countries. Our findings highlight the role that frozen foods may play in the spread of this pathogen, and the relevance of serotypes 1/2b and 4b.

ABSTRACT Laboratory diagnosis of human leptospirosis usually relies on indirect methods exploring specific immune response. Isolation and identification of the involved strains are cumbersome, but can provide biological resources for pathogenic studies and relevant information for guiding prevention and control measures. The aim of the research we are hereby reporting was the characterization of Leptospira isolates obtained from humans and the environment in Uruguay. Blood cultures were performed from early samples of 302 Uruguayan patients, mainly rural workers, and from 36 water samples taken from their living or working environments. Eight human isolates and seven environmental isolates were obtained and analyzed by end point Polymerase Chain Reaction (PCR), Multilocus Variable Number of Tandem Repeat Analysis (MLVA) and other molecular methods. Human isolates corresponded to several serogroups and serovars of Leptospira interrogans and Leptospira kirschneri species, probably reflecting the infection with similar involved Leptospira species and serovars of an extended animal reservoir in rural settings of the country, mostly dedicated to meat and dairy production. Culture-positive patients were older than usually affected workers, and presented signs and symptoms of severe illness. A high organic and circulating bacterial burden may explain an easier positive result from these workers’ samples. Environmental isolates were mainly identified as Leptospira biflexa strains, with a single L. meyeri isolate of uncertain significance.

Introducción: la leptospirosis es una enfermedad febril, aguda, que presenta manifestaciones clínicas variadas. Esto dificulta o retarda el diagnóstico clínico, por lo cual es útil disponer de métodos de laboratorio adecuados para orientar el manejo inicial de estos pacientes. Objetivo: evaluar un procedimiento de inmunofluorescencia para detectar IgM (IF-IgM) de elaboración propia aplicado al diagnóstico temprano de leptospirosis. Material y método: se analizaron por IF-IgM y microaglutinación (MAT) (tomada como estándar de referencia) sueros obtenidos de pacientes con sospecha clínica de leptospirosis. La sensibilidad y especificidad de la IF-IgM versus MAT se establecieron utilizando una tabla de doble entrada. La concordancia entre dos observadores se determinó por el test Kappa. Resultados: de 161 muestras precoces analizadas, 97 sueros correspondieron a pacientes con infección aguda confirmada por MAT y 64 sin infección. La sensibilidad y especificidad de la IF-IgM con sueros de fase aguda fueron 79% y 100%, respectivamente. El índice Kappa fue 1. Conclusiones: la IF-IgM aparece como una herramienta útil para el diagnóstico temprano de pacientes con leptospirosis. No requiere el manejo de bacterias viables, puede realizarse en laboratorios que cuenten con microscopio de luz ultravioleta, se necesita una sola muestra de suero y el resultado está listo en tres a cuatro horas. En cuanto a las desventajas, no identifica los serovares involucrados y un resultado negativo no descarta la infección. Teniendo en cuenta esto último es obligatorio analizar por MAT una segunda muestra de suero obtenida a los 10-20 días de la primera para descartarla o confirmarla.

Resumo Introdução: a leptospirose é uma doença febril, aguda, com diferentes manifestações clínicas o que dificulta ou atrasa o diagnóstico clínico. Por essa razão é importante contar com métodos de laboratório adequados para orientar o manejo inicial dos pacientes. Objetivo: avaliar uma técnica de imunofluorescencia para detectar IgM (IF-IgM) para o diagnóstico precoce de leptospirose. Material e método: foram analisados por IF-IgM e microaglutinaçao (MAT) (usada como padrão de referencia) soros de pacientes com suspeita clínica de leptospirose. A sensibilidade e a especificidade da IF-IgM comparada com MAT foi determinada usando uma tabela de dupla entrada. A concordância entre dois observadores foi determinada usando o teste Kappa. Resultados: Cento sessenta e uma amostras da fase precoce foram analisadas, sendo 97 de pacientes com infecção aguda confirmada por MAT e 64 sem infecção. A sensibilidade e a especificidade da IF-IgM com soros obtidos na fase aguda foram 79% e 100%, respectivamente. O índice Kappa foi igual a 1. Conclusões: a técnica IF-IgM que desenvolvemos, pode ser uma ferramenta útil para o diagnóstico precoce de pacientes com leptospirose. Não é necessário manipular bactérias viáveis, pode ser realizada em laboratórios que tenham microscópio com luz ultravioleta, somente é necessária uma amostra de soro e o resultado está disponível em três-quatro horas. Com relação às desvantagens, esta técnica não identifica os serovares e um resultado negativo não descarta a presença de infecção. Considerando estas desvantagens, é obrigatório analisar por MAT uma segunda amostra de soro obtida 10 a 20 dias depois da primeira para descartar ou confirmar a infecção.

Abstract Introduction: leptospirosis is a severe febrile disease, with a variety of clinical presentations. This results in the delay of clinical diagnosis, or in difficulties achieving it, so having the appropriate laboratory tests may deem useful to guide the initial care of these patients. Objective: to evaluate a self-made immunofluorescence technique for the detection of IgM antibodies (IF-IgM) in the early diagnosis of leptospirosis. Method: serum samples obtained from patients with a clinical suspicion of leptospirosis were analysed by IF-IgM antibodies and the microagglutination test (MAT) . Sensitivity and specificity of IF-IgM versus MAT were determined using a double entry table. Agreement between two observers was determined by the Kappa test. Results: out of one hundred and sixty one early samples analysed, 97 serum samples corresponded to patients with a confirmed acute infection by MAT and 64 evidenced no infection. Sensitivity and specificity of IF-IgM assays in the acute phase were 79% and 100% respectively. The Kappa value was 1. Conclusions: IF-IgM seems to be a useful tool for the early diagnosis of patients with leptospirosis. It does not need viable bacteria to be handled; it may be applied in laboratories equipped with ultraviolet light microscopes, a single serum sample is needed and the result is seen three to four hours later. As to its disadvantages, it fails to identify the involved serovars and a negative result does not discard infection. Bearing the latter into account, the MAT test is mandatory in a second serum sample obtained 10-20 days after the first one, to either discard or confirm the disease.

Hemolytic uremic syndrome (HUS) is a disorder characterized by the presence of the classic triad: microangiopathic hemolytic anemia, thrombocytopenia and acute renal injury. HUS without acute renal failure can be confused with other hematologic diseases. An infantile HUS caused by a Shiga-toxin-producing Escherichia coli (STEC) O145 strain carrying genotype stx2, ehxA, eae subtype ?1 is herein reported. The infant did not require dialysis during the acute stage of HUS, evolved favorably, maintained normal blood pressure and normal renal function and had no recurrence until the last control. This could be due to several factors, such as the characteristics of infecting STEC strain and a reduction in host susceptibility to renal injury. This report highlights the regional participation of non-O157 STEC in childhood diseases and the importance of performing active surveillance for all forms of HUS.

El síndrome urémico hemolítico (SUH) es una afección caracterizada por la presencia de la tríada clásica: anemia hemolítica microangiopática, trombocitopenia y compromiso renal agudo. Los casos de SUH sin insuficiencia renal pueden confundirse con otras enfermedades hematológicas. Presentamos un caso de SUH pediátrico causado por una cepa de Escherichia coli productora de toxina Shiga Shiga-toxin-producing Escherichia coli (STEC) O145 con el genotipo stx2, ehxA, eae subtipo ?1. El niño no requirió diálisis durante la etapa aguda del SUH, evolucionó favorablemente y no tuvo recurrencias hasta el último control; además, mantuvo cifras normales de presión arterial y función renal normal. Esto puede deberse a varios factores: características de la cepa STEC infectante y susceptibilidad del hospedero al daño renal, entre otros. Este hallazgo destaca la participación regional de STEC no-O157 en enfermedades de la infancia y la importancia de realizar una vigilancia activa de todas las formas de SUH.

Infections due to Shigella usually remain localized to the digestive tract and are self-limited. Bacteremia is a potentially lethal complication that can occur in immunocompromised patients. We describe two episodes of bacteremia caused by Shigella in two adults with AIDS. In both patients, S. flexneri was recovered from stool and blood samples. The isolates belonged to serotype 6, were resistant only to trimethoprim-sulfamethoxazole and showed a similar band profile by pulsed-field gel electrophoresis. Patients received prolonged antimicrobial treatment with a favorable outcome. There were no cases of diarrhea in other individuals admitted to the emergency room. We hypothesized that patient No. 2 was infected at the hospital from patient No. 1. However, we could not establish the way of transmission. Although rare, it is important to take into account the possible occurrence of bacteremia due to Shigella or other bacterial enteropathogens in immunocompromised patients with diarrhea.

Las infecciones por Shigella spp., en general, permanecen localizadas en el tracto digestivo y tienen una evolución autolimitada. La bacteriemia es una complicación potencialmente letal que ocurre en pacientes con algún tipo de inmunocompromiso. Presentamos dos casos de bacteriemia causadas por Shigella en dos adultos con SIDA. En ambos pacientes, se recuperó Shigella flex-neri en muestras de deposiciones y sangre. Los aislados correspondieron al serotipo 6, fueron resistentes sólo a cotrimoxazol y mostraron un perfil de bandas similar por PFGE. Los pacientes recibieron tratamiento antimicrobiano prolongado y la evolución fue favorable. No se registraron otros casos de diarrea en individuos admitidos en el servicio de emergencia. La hipótesis fue que el paciente 2 adquirió la infección en el hospital a partir del paciente 1. Sin embargo, no pudimos establecer el modo de transmisión. Aunque poco frecuente, es importante tener presente la ocurrencia de bacteriemia por Shigella spp. o por otros enteropatógenos bacterianos en pacientes inmunocomprometidos con diarrea.

ABSTRACT STEC strains can infect extra-intestinal sites such as the human urinary tract and sometimes cause severe complications. We report two cases of urinary tract infection caused by STEC in two elderly women with comorbidities. Although both strains belonged to the 0157:H7 serotype and carried genes associated with severe illness, none of the patients developed hemolytic uremic syndrome (HUS). These findings provide additional evidence for the presence of these agents in our country and in the region, and highlight the need to maintain an active surveillance system of HUS cases, placing special emphasis on the study of other sites of infection in patients with non-diarrheal HUS.

En los seres humanos, las cepas STEC pueden producir infección en sitios extraintestinales, como el tracto urinario, y causar complicaciones graves. Comunicamos dos casos de infección urinaria por STEC en dos mujeres ancianas con comorbilidades. Aunque ambas cepas correspondieron al serotipo 0157:H7 y portaban los genes asociados con enfermedad grave, ninguna de las pacientes desarrolló síndrome urémico hemolítico (SUH). Estos hallazgos constituyen una evidencia adicional de la presencia de estos agentes en nuestro país y en la región, y destacan la necesidad de mantener un sistema activo de vigilancia, con especial énfasis en el estudio de otros sitios de Infección en pacientes que presentan SUH no asociado a diarrea.

Leptospira spp. are delicate bacteria that cannot be studied by usual microbiological methods. They cause leptospirosis, a zoonotic disease transmitted to humans through infected urine of wild or domestic animals. We studied the incidence of this disease in the Uruguayan population, its epidemiologic and clinical features, and compared diagnostic techniques. After examining 6,778 suspect cases, we estimated that about 15 infections/100,000 inhabitants occurred yearly, affecting mainly young male rural workers. Awareness about leptospirosis has grown among health professionals, and its lethality has consequently decreased. Bovine infections were probably the principal source of human disease. Rainfall volumes and floods were major factors of varying incidence. Most patients had fever, asthenia, myalgias or cephalalgia, with at least one additional abnormal clinical feature. 30-40% of confirmed cases presented abdominal signs and symptoms, conjunctival suffusion and altered renal or urinary function. Jaundice was more frequent in patients aged > 40 years. Clinical infections followed an acute pattern and their usual outcome was complete recovery. Laboratory diagnosis was based on indirect micro-agglutination standard technique (MAT). Second serum samples were difficult to obtain, often impairing completion of diagnosis. Immunofluorescence was useful as a screening test and for early detection of probable infections.

El género Leptospira comprende bacterias delicadas, que requieren métodos de estudio especiales. Causan una zoonosis transmitida a los seres humanos por la orina de animales domésticos o silvestres. Estudiamos la incidencia de la leptospirosis en la población humana de Uruguay, sus características epidemiológicas y clínicas, y comparamos técnicas diagnósticas. Tras examinar 6778 casos sospechosos, estimamos que anualmente ocurren unas 15 infecciones/100.000 habitantes, principalmente en trabajadores rurales jóvenes de sexo masculino. El nivel de alerta sobre la leptospirosis ha avanzado en el personal sanitario, y su letalidad ha descendido. La enfermedad humana se asocia principalmente con las infecciones bovinas. Lluvias e inundaciones influyen sobre su incidencia. La mayoría de los pacientes experimentaron fiebre, astenia, mialgias o cefaleas, con al menos un síntoma o signo adicional. 30 a 40% presentaban sintomatología abdominal, hiperemia conjuntival o alteraciones nefro-urológicas. La ictericia era más frecuente en mayores de 40 años. La enfermedad era habitualmente aguda y la recuperación completa. El diagnóstico de laboratorio se basó en la técnica de microaglutinación (MAT), con cepas vivas de distintos serovares. Se requieren segundas muestras de suero, de difícil obtención. El test de inmunofluorescencia se reveló útil para screening y detección precoz de la infección.

Introducción: la infección por Estreptococo grupo B (EGB) puede afectar gravemente a la madre y al feto durante la gestación y al recién nacido luego del parto. Actualmente, el diagnóstico de colonización durante el embarazo se realiza por métodos microbiológicos a partir de exudados vaginorrectales. Objetivo: desarrollar métodos rápidos y de bajo costo para la detección del antígeno grupo específico de EGB en exudados vaginorrectales. Material y método: se utilizaron dos cepas de EGB, una autóctona (IH23) y la cepa de referencia O90R, que solo expresa el polisacárido específico de grupo. Para cada una se preparó un antisuero policlonal que se utilizó para el desarrollo de un test inmunocromatográfico y uno de aglutinación de látex. Como controles se emplearon cultivos bacterianos, polisacáridos purificados de EGB y muestras vaginorrectales. Resultados: los límites de detección obtenidos para la inmunocromatografía fueron de 210 µg/ml y 50 µg/ml para los polisacáridos purificados de sobrenadante y pared, respectivamente, no lográndose detectar antígenos de EGB en las muestras clínicas analizadas. El límite de detección del látex fue 65 µg/ml frente al polisacárido purificado de sobrenadante de cultivo de IH23 y 6,5 x 107 UFC/ml de IH23. La sensibilidad y especificidad para el látex fue de 30% y 90%, respectivamente. Conclusiones: los métodos desarrollados no alcanzaron el límite de detección requerido para su aplicación en muestras clínicas. Esto concuerda con lo descripto en la bibliografía para ensayos rápidos basados en reacciones antígeno-anticuerpo y muestra la necesidad de agregar pasos previos de extracción y concentración o mejorar la calidad de los reactivos inmunológicos empleados.

Resumo Introdução: a infecção por Estreptococo grupo B (EGB) pode afetar gravemente a mãe e o feto durante a gravidez e o recém nascido imediatamente depois do parto. Atualmente, o diagnóstico de colonização durante a gestação é feita por métodos microbiológicos empregando exsudados vaginorretais. Objetivo: desenvolver métodos rápidos de baixo custo para a detecção do antígeno grupo específico de EGB em exsudados vaginorretais. Material e método: foram utilizadas duas cepas de EGB, uma autóctone (IH23) e a cepa de referência O90R, que expressa somente o polissacarídeo específico do grupo. Para cada uma foi preparado um antisoro policlonal utilizado para o desenvolvimento de um teste imunocromatográfico e um de aglutinação de látex. Foram empregados cultivos bacterianos, polissacarídeos purificados de EGB e amostras vaginorretais como controles. Resultados: os limites de detecção obtidos para a imunocromatografia foram de 210 µg/ml e 50 µg/ml para os polissacarídeos purificados de sobrenadante e parede, respectivamente, no sendo possível detectar antígenos de EGB nas amostras clínicas analisadas. O limite de detecção do látex foi 65 µg/ml quando comparado com o polissacarídeo purificado de sobrenadante de cultivo de IH23 y 6,5 x 107 UFC/ml de IH23. A sensibilidade e especificidade para o látex foi de 30% y 90%, respectivamente. Conclusões: os métodos desenvolvidos não alcançaram o limite de detecção requerido para sua aplicação em amostras clínicas. Estes resultados são similares aos dados descritos na bibliografia para ensaios rápidos baseados em reações antígeno-anticorpo e mostram a necessidade de agregar passos prévios de extração e concentração ou melhorar a qualidade dos reativos imunológicos utilizados.

Summary Introduction: group B streptococcal infection (GBS) may seriously affect mother and fetuses during pregnancy, and the newborn after delivery. Today, diagnosis of colonization during pregnancy is done by means of microbiological methods of vaginal and rectal exudates. Objective: to develop fast and low cost methods to detect the GBS specific group antigen in vaginal-rectal exudates. Method: we used two EGB strains, one of the (IH23) autochthonous and the reference strain O90R, that only expresses the group specific polysaccharide. We prepared a polyclonal antiserum for each one of them which was used to conduct an immunochromatographic test and a latex agglutination test. We used bacterial culture, EGB purified polysaccharides and vagina,-rectal samples as control. Results: detection limits obtained for the immunochromatographic test were 210 µg/ml and 50 µg/ml for purified polysaccharides and cell wall, respectively, there being no EGB antigens detected in the clinical samples analyzed. Latex detection limit was 65 µg/ml compared to purified polysaccharides of IH23 culture supernatant and 6,5 x 107 UFC/ml of IH23. Sensitivity and specificity for latex was 30% and 90% respectively. Conclusions: the methods used failed to reach the detection limit required for its application in our clinical samples. This agrees with what is described in bibliography about quick tests based on antigen-antibody reactions and indicated the need to add previous extraction and concentration steps or to improve the quality of the immunologic reagents used.