Seroprevalencia de anticuerpos anti-Mycoplasma pneumoniae en niños menores de 12 años / Seroprevalence of anti-Mycoplasma pneumoniae antibodies in otherwise healthy children

Resumen: Introducción: La infección por Mycoplasma pneumoniae (Mypn) podría estar ocurriendo a edades más tempranas, debido a fenómenos sociales como concurrencia a centros de cuidado diurno en forma más frecuente y precoz. Objetivo: estimar la prevalencia de anticuerpos anti-Mypn en niños de 0-12 años, y explorar si la edad, asistencia a centro de cuidados diurnos/escuela, hacinamiento o convivencia con niños incrementan el riesgo de seropositividad. Pacientes y Método: Estudio transversal incluyendo niños de 0-12 años de edad que requirieron extracciones de sangre para control, por lo demás sanos. En todos los casos se consignaron las variables mencionadas y se determinó IgG anti-Mypn mediante enzimoinmunoanálisis. Se evaluó la asociación entre predictores y seropositividad en un modelo de regresión logística. Resultados: Se incluyeron 232 pacientes (edad promedio 56,4 ± 40,0 meses). El 56,9% concurría a centro de cuidado diurno/escuela, 63,8% convivían con menores de 12 años y 15,9% presentaban hacinamiento. El 14,6% presentaba anticuerpos anti-Mypn. Los niños seroposi- tivos no mostraron diferencias significativas con aquellos seronegativos en relación a edad (63,1 ± 40,7 vs. 55,4 ± 41,3 meses), escolaridad (64,7% vs 55,5%), hacinamiento (14,7% vs 14,9%), ni con vivencia con menores (64,7% vs 63,6%). La edad tampoco se mostró como predictor independiente de seropositividad en el modelo multivariado. Conclusión: La prevalencia de anticuerpos anti-Mypn fue 14,6%. La edad no fue predictor de seropositividad.

Abstract: Introduction: Mycoplasma pneumoniae (Mypn) infection could be occurring at an earlier age due to social pheno mena such as attending daycare centers more frequently and earlier than decades ago. Objective: to estimate the prevalence of anti-Mypn antibodies in children aged 0-12 years, and to explore whether age, attendance to daycare center/school, overcrowding or the presence of children aged below 12 years in the households increase the risk of seropositivity. Patients and Method: Cross-sectional stu dy including healthy children aged 0-12 years which required blood draws for routine laboratory tests. In all cases, the aforementioned variables were recorded and anti-Mypn IgG was determined by enzyme immunoassay. The association between predictors and seropositivity was assessed in a logistic regression model. Results: We included 232 patients (average age 56.4 ± 40.0 months). 56.9% attended a daycare center/school, 63.8% co-habited with children under 12 years old, and 15.9% lived in overcrowded households. The prevalence of anti-Mypn antibodies was 14.6%. There were no significant differences between seropositive and seronegative children regarding age (63.1 ± 40.7 vs. 55.4 ± 41.3 months), school/day-care attendance (64.7% vs. 55.5%), overcrowding (14.7% vs. 14.9%), or co-habiting with children (64.7% vs. 63.6%). Age was not an independent predictor of seropositivity in the multivariate model. Conclusion: The prevalence of anti-Mypn antibodies in children was 14.6% and age was not a predictor of seropositivity.

Serosurvey on rickettsiae of the spotted fever group and Rickettsia bellii among dogs in the state of Goiás, Brazil / Inquérito sorológico sobre riquétsias do grupo da febre maculosa e Rickettsia bellii entre cães no estado de Goiás, Brasil

Abstract The purpose of this study was to do a serological survey on three rickettsial species: Rickettsia rickettsii and Rickettsia parkeri, two species of the spotted fever group (SFG) that are considered to be great importance for public health; and Rickettsia bellii, a species of unknown pathogenicity that infects a variety of human-biting ticks. Serum samples from 273 dogs were tested using the indirect immunofluorescence assay (IFA). A total of 52 samples (19.04%) were seropositive for at least one of the three Rickettsia spp. antigens. Thirty-eight (73.07%), twelve (23.07%) and one (1.92%) of these dogs showed homologous reactions to R. bellii, R. rickettsii and R. parkeri, respectively. Our results showed that the seroprevalence of Rickettsia spp. was relatively low. However, the positive serological tests indicated that these dogs had become infected by these agents at some point in their lives. Lastly, our study adds to the previous knowledge on the epidemiology of rickettsiosis in the state of Goiás by doing the first record of detection of anti-R. rickettsii, R. parkeri and R. bellii antibodies by IFA among dogs, thus indicating that these agents may be circulating in the dog population analyzed.

Resumo O objetivo deste estudo foi realizar um levantamento sorológico para três espécies de rickettsias: Rickettsia rickettsii e Rickettsia parkeri, duas espécies do grupo da febre maculosa (GFM) consideradas de grande importância para a saúde pública; e Rickettsia bellii, uma espécie de patogenicidade desconhecida que infecta uma variedade de carrapatos que parasitam seres humanos. Amostras de soro de 273 cães foram testadas, usando-se a técnica de reação de imunofluorescência indireta (RIFI). O total de 52 amostras (19,04%) foram soropositivas para pelo menos um dos três antígenos de Rickettsia spp. Trinta e oito (73,07%), doze (23,07%) e um (1,92%) desses cães apresentaram reações homólogas à R. bellii, R. rickettsii e R. parkeri, respectivamente. Esses resultados demonstraram uma baixa soroprevalência para Rickettsia spp. No entanto, as amostras positivas indicam que esses cães foram infectados por esses agentes em algum momento de suas vidas. Por fim, este estudo contribui para o conhecimento sobre a epidemiologia das rickettsioses no estado de Goiás, realizando a primeira detecção de anticorpos anti-Rickettsia rickettsii, R. parkeri e R. bellii pela RIFI em cães, indicando que esses agentes podem estar circulando na população canina analisada.

Detection of tick-borne rickettsial pathogens in naturally infected dogs and dog-associated ticks in Medellin, Colombia / Detecção de patógenos rickettsiais transmitidos por carrapatos em cães naturalmente infectados e carrapatos associados a cães em Medellin, Colômbia

Abstract Tick-borne rickettsial pathogens (TBRP) are important causes of infections in both dogs and humans. Dogs play an important role as a biological host for several tick species and can serve as sentinels for rickettsial infections. Our aim was to determine the presence of TBRP in dogs and in dog-associated ticks and their potential risk to human diseases in Medellin, Colombia. DNA for E. canis (16S rRNA and dsb) and A. platys (groEl) was detected in 17.6% (53/300) and 2.6% (8/300) of dogs, respectively. Antibodies against Ehrlichia spp. 82 (27.3%) and Anaplasma spp. 8 (2.6%) were detected in dogs. Antibody reactivity against both agents were found in 16 dogs (5.3%). Eight dogs showed antibody for Rickettsia spp. with titers that suggest 3 of them had a probable exposure to R. parkeri. Rhipicephalus sanguineus s.l. (178/193) was the main tick in dogs, followed by R. microplus (15/193). The minimum infection rates (MIR) in R. sanguineus were 11.8% for E. canis and 3.4% for A. platys. E. canis and A. platys are the main TBRP infecting dogs and ticks and R. sanguineus s.l. is likely involved in the transmission of both agents. Interestingly, we found serological evidence of exposure in dogs for spotted fever group rickettsiae.

Resumo As riquétsias transmitidas por carrapatos (RTC) são causas importantes de infecção em cães e humanos. Os cães exercem um papel essencial como hospedeiros biológicos para diversas espécies de carrapatos, assim como podem ser úteis como sentinelas de infecções por riquétsias. O intuito deste estudo foi determinar a presença de RTC em cães, assim como em seus carrapatos, para determinar o risco potencial de doença humana em Medellín, Colômbia. DNA de Ehrlichia canis (16S rRNA e dsb) e Anaplasma platys (groEl) foi detectado em 17,6% (53/300) e 2,6% (8/300) dos cães, respectivamente. Anticorpos contra Ehrlichia spp. (82; 27,3%) e Anaplasma spp. (8; 2,6%) foram detectados nos cães. Reatividade de anticorpos contra ambos patógenos (Ehrlichia e Anaplasma) foi detectada em 16 cães (5,3%). Oito animais apresentaram anticorpos contra Rickettsia spp., e 3 deles sugerem uma provável exposição a Rickettsia parkeri. Rhipicephalus sanguineus s.l. (178/193) foi a principal espécie de carrapatos, seguida de R. microplus (15/193). A taxa de infecção mínima em R. sanguineus foi 11,8% para E. canis e 3,4% para A. platys. E. canis e A. platys são as principais RTC que infectam cãese R. sanguineus s.l. provavelmente está envolvido na transmissão de ambos os agentes. É evidente, porém, a exposição sorológica dos cães a riquétsias do grupo da febre maculosa.

Spotted fever group rickettsial infection in dogs and their ticks from domestic-wildlife interface areas in southeastern Brazil / Infecção por rickettsias do Grupo da Febre Maculosa em cães e carrapatos de áreas de interface doméstico-silvestre no sudeste Brasileiro

Abstract Rickettsia rickettsii is the causative agent of Brazilian spotted fever (BSF), for which humans and dogs are both susceptible. Dogs are sentinels in serological surveys, however, canine disease is rarely reported. Therefore, we aimed to evaluate natural infection by spotted fever group (SFG) Rickettsia spp. in dogs and ticks collected from domiciles close to forest fragments, featuring domestic-wildlife interface areas. Samples from 115 dogs and 135 ixodids were assessed by polymerase chain reactions (PCR) targeting the gltA gene for Rickettsia spp. and the ompA gene for the SFG rickettsial species. One dog (0.87%; 1/115) was positive for R. rickettsii. This dog presented nonspecific laboratory and clinical abnormalities (thrombocytopenia, hyperproteinemia, lymph node enlargement, emaciation, anorexia, and lethargy). Rickettsia parkeri was identified in 2.96% (4/135) of the ticks (Amblyomma sculptum, A. aureolatum, and Rhipicephalus sanguineus). This study confirmed the presence of SFG bacteria in non-endemic and preserved locations, where domestic and wild populations interact. We reinforce the fact that the dog is susceptible to natural R. rickettsii infection. Although this is a rare finding, preventive measures should be taken against BSF in the studied areas. Finally, R. parkeri infection is possibly being demonstrated in A. sculptum for the first time.

Resumo Rickettsia rickettsii é o agente causador da Febre Maculosa Brasileira (FMB), doença na qual humanos e cães são susceptíveis. Os cães são sentinelas nos inquéritos sorológicos, contudo, a doença canina é raramente descrita. Assim sendo, objetivou-se avaliar a infecção natural por Rickettsia spp. do Grupo da Febre Maculosa (GFM) em cães e carrapatos obtidos de domicílios próximos a fragmentos de mata, caracterizando áreas de interface doméstico-silvestre. Amostras de 115 cães e 135 ixodídeos foram avaliadas pela reação em cadeia da polimerase (PCR) tendo como alvo o gene gltA de Rickettsia spp. e o gene ompA das espécies do GFM. Um cão (0,87%; 1/115) foi positivo para R. rickettsii. Este animal apresentou alterações clínicas e laboratoriais inespecíficas (trombocitopenia, hiperproteinemia, linfonodos edemaciados, emagrecimento, anorexia e letargia). Rickettsia parkeri foi identificada em 2,96% (4/135) dos carrapatos (Amblyomma sculptum, A. aureolatum e Rhipicephalus sanguineus). Este estudo confirmou a presença de bactérias do GFM em locais preservados e não endêmicos, onde populações domésticas e silvestres interagem. Reforçamos o fato do cão ser susceptível à infecção natural por R. rickettsii. Embora este seja um achado raro, medidas preventivas devem ser tomadas contra a FMB nas áreas estudadas. Em última análise, a infecção por R. parkeri possivelmente está sendo demonstrada pela primeira vez em A. sculptum.

Serological occurrence for tick-borne agents in beef cattle in the Brazilian Pantanal / Ocorrência sorológica para agentes transmitidos por carrapatos em bovinos de corte no Pantanal Brasileiro

Abstract This study investigated the seropositivity for five different tick-borne agents, namely Anaplasma marginale, Babesia bovis, Babesia bigemina, Coxiella burnetii, Anaplasma phagocytophilum, and Trypanosoma vivax in beef cattle in the Brazilian Pantanal. The serum samples collected from animals (200 cows; 200 calves) were used in indirect enzyme-linked immunosorbent assays (iELISA) to detect IgG antibodies against A. marginale, B. bovis, B. bigemina, and T. vivax, and Indirect Fluorescent Antibody Test (IFAT) for detecting IgG antibodies against C. burnetii and A. phagocytophilum. No correlation was observed between seropositivity for C. burnetii and A. phagocytophilum with other agents whereas moderate correlation was observed for A. marginalexB. bigemina x B. bovis. Cows were more seropositive for T. vivax whereas calves were more seropositive for B. bovis and B. bigemina. The highest number of seropositive animals by a single agent was observed for T. vivax (15.2%). Co-seropositivity for T. vivax + A. marginale was higher in cows (25.5%) and for T. vivax + B. bovis + B. bigemina + A. marginale was higher in calves (57.5%). The high seropositivity correlation for A. marginale x B. bovis x B. bigemina is probably due to the presence of the tick biological vector, Rhipicephalus microplus, in the studied farms. Common transmission pathways, mediated by hematophagous dipterans and fomites, may explain the high co-seropositivity of cows for A. marginale and T. vivax. Low seropositivity to C. burnetii is probably due to the type of breeding system employed (extensive). Seropositivity for A. phagocytophilum in only one animal suggests the occurrence of a cross-serological reaction with another agent of the genus Anaplasma.

Resumo Este estudo teve como objetivo determinar a co-soropositividade para agentes transmitidos por carrapatos, como Anaplasma marginale, Babesia bovis, Babesia bigemina, Coxiella burnetii, Anaplasma phagocytophilum, e Trypanosoma vivax em bovinos de corte do Pantanal Brasileiro. Amostras de soro foram colhidas de 400 animais (200 vacas; 200 bezerros) e submetidas a Ensaios Imunoenzimáticos Indiretos (iELISA) para detecção de anticorpos IgG anti- A. marginale, anti- B. bovis, anti- B. bigemina e anti- T. vivax, e à Reação de Imunofluorescência Indireta (RIFI) para detecção de anticorpos IgG anti -C. burnetii e anti- A. phagocytophilum. Ausência de correlação foi vista entre os animais soropositivos para C. burnetii e A. phagocytophilum com os outros agentes e correlação moderada ocorreu entre A. marginale x B. bigemina x B. bovis. Vacas foram mais soropositivas que bezerros para T. vivax e bezerros mais soropositivos que vacas para B. bovis e B. bigemina. Maior número de animais soropositivos para um único agente foi visto para T. vivax (15,2%). Vacas demonstraram maior co-soropositividade para T. vivax + A. marginale (25,5%) e bezerros para T. vivax + B. bovis + B. bigemina + A. marginale (57,5%). A alta correlação entre a soropositividade para A. marginale x B. bovis x B. bigemina é provavelmente devida à presença do vetor biológico, o carrapato Rhipicephalus microplus, nas fazendas estudadas. As vias de transmissão comuns, mediadas por dípteros hematófagos e fômites, podem explicar a alta co-soropositividade das vacas para A. marginale e T. vivax. A baixa soropositividade para C. burnetii é provavelmente devida ao tipo de sistema de criação empregado (extenso). A soropositividade para A. phagocytophilum em apenas um animal sugere a ocorrência de reação sorológica cruzada com outro agente do gênero Anaplasma.

Seroprevalencia de leptospirosis en grupos humanos en riesgo por factores ambientales, laborales o sociales / Seroprevalence of leptospirosis in human groups at risk due to environmental, labor or social conditions

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.

Serosurveillance for vaccine-preventable diseases: A look inside the pertussis experience / Serovigilancia de enfermedades prevenibles por vacunación: una mirada desde la experiencia con la tosferina

Abstract Introduction: Serological surveillance (serosurveillance) provides the most direct measure of herd immunity of vaccine-preventable diseases. Little is known about the opportunities and challenges of serosurveillance experiences, particularly pertussis. Objective: To describe the process of serosurveillance for vaccine-preventable diseases with an emphasis on the experience of pertussis in the metropolitan area of Antioquia (Valle de Aburrá) in 2015 and 2016 and analyze the contributions and challenges for its sustainability. Materials and methods: We described the planning and conduction of serosurveillance of pertussis antibodies of mothers and in the umbilical cord at the time of delivery in eight hospitals based on random sampling and their capacity to advance the serosurveillance periodically. We compared the contributions and the challenges of this experience with other probabilistic and non-probabilistic programs. Results: We achieved the participation of hospitals and mothers respecting the delivery care process. We established a serum bank following ethical and technical guidelines. This program based on the random selection of hospitals and mothers has enabled the estimation of antibodies prevalence in mothers and in the umbilical cord, which has been possible given the high coverage of hospital care during childbirth at a lower cost and fewer risks than a population-based survey in conflictive areas. The main challenges for the sustainability of this program are the creation of stable jobs and access to funding and legal and methodological long-term frameworks. Conclusions: Hospital serosurveillance as described is an option to monitor the impact of vaccination on the population. Our experience could be reproduced in other regions under similar conditions if the above-mentioned challenges are solved.

Resumen Introducción. La vigilancia serológica es la forma más directa de medir la inmunidad de rebaño frente a las enfermedades prevenibles por vacunación. Poco se sabe acerca de las oportunidades y los desafíos de las experiencias de serovigilancia, en general y, específicamente, la de la tosferina. Objetivo. Describir el proceso de serovigilancia de enfermedades prevenibles por vacunación con énfasis en la experiencia en el caso de la tosferina en el área metropolitana de Antioquia (Valle de Aburrá) en el 2015 y el 2016 y analizar lo que dicha experiencia ha aportado y los desafíos que persisten para su sostenibilidad. Materiales y métodos. Se describió el proceso de planeación y el desarrollo de la serovigilancia de tosferina en el momento del parto en ocho hospitales seleccionados al azar, así como la capacidad para adelantar el programa de manera periódica. Se compararon los aportes y los desafíos en el curso de esta experiencia con los de otros programas poblacionales probabilistas e institucionales no probabilistas. Resultados. Se logró la participación de los hospitales y de las madres con pleno respeto del proceso de atención del parto, y se conformó un banco de sueros siguiendo lineamientos éticos y técnicos. El programa permitió estimar la prevalencia de anticuerpos en la madre y en el cordón umbilical, lo que se facilitó por la alta cobertura de atención hospitalaria del parto, a un menor costo y menos riesgos que los programas poblacionales en zonas conflictivas. Los principales desafíos para la sostenibilidad del programa son la estabilidad laboral del personal de salud, así como normas y una financiación de largo plazo. Conclusiones. La serovigilancia hospitalaria es una opción para monitorizar el impacto poblacional de la vacunación. Esta experiencia se podría extender a otras regiones en condiciones similares si se resuelven los retos mencionados.

Leprosy: prevalence and factors associated with seropositivity for anti-NDO-LID antibodies in children under 15 years of age

Abstract: Background: A high prevalence of leprosy among children under 15 years of age indicates the need to implement actions to prevent new cases of the disease. Serological tests have been developed with the aim of helping to control the disease by indicating, through seropositivity, the presence of infection. Objective: To analyze the prevalence and factors associated with seropositivity rate for anti-NDO-LID antibodies in children under 15 years of age, contacts of leprosy patients. Method: We performed a cross-sectional study with 210 children under 15 years old of age. Of them, 50 were household contacts and 160 were neighborhood contacts living in the municipality of Cuiabá, state of Mato Grosso, in 2016. The data were obtained from interviews and the NDO-LID rapid test during home visits from February to July 2016. For the analysis, we used Poisson regression and prevalence ratio. Results: Seropositivity in contacts was 6.2%. Variables associated with seropositive tests included sex (PR = 1.05; 95% CI: 1.01 - 1.08), race/skin color (PR = 0.95; 95% CI: 0.90 - 0.99), residence area (PR = 1.05; 95% CI: 1.01 - 1.09), and number of people per household (PR = 1.06; 95% CI: 1.02 - 1.08). Study Limitations: The small sample size, besides leading to wide confidence intervals, may have been a limitation for the identification of associated factors. Conclusions: The prevalence of seropositivity was high. Variables associated with NDO-LID seropositivity included female sex, not to be brown skinned, live in urban areas, and live with five or more people.

Epidemiological relevance of dogs for the prevention of Toxoplasma gondii, Neospora caninum and Leptospira spp / Relevância epidemiológica de cães para a prevenção de Toxoplasma gondii, Neospora caninum e Leptospira spp

Abstract This study evaluated the seroprevalence of Toxoplasma gondii, Neospora caninum and Leptospira spp. in dogs from Foz do Iguaçu, Paraná, Brazil. Indirect immunofluorescent antibody test was used to detect antibodies anti-T.gondii and anti-N. caninum. Immunoenzymatic assay and microscopic serum agglutination were used for screening antibodies anti-T.gondii and anti-Leptospira spp., respectively. The results were: 67.02% of the samples reactive for T.gondii and 1.38% for N. caninum, both without statistically significant variables. For Leptospira spp. the results indicated seroprevalence of 23.11%. The analysis of the variables without distinction of serovar showed association for intrinsic characteristics as breed, age, nutritional status and dog category. The extrinsic variables as city region and access to the street presented association (p<0.05). The most prevalent serovars were: Canicola 59.47%; Bratislava 13.07% and Butembo 15.68%. Variables that make up the adjusted multiple analysis model using Leptospira spp. were: age, breed and nutritional status; serovar Canicola, sex, nutritional status and area (p<0.05); serovar Bratislava, lymphadenomegaly and presence of fleas (p<0.05). Given the results obtained, dogs can be used as sentinels for toxoplasmosis and leptospirosis in Foz do Iguaçu and other cities with similar outcomes. In addition, preventive measures should be taken by health authorities because they are zoonoses and humans are also at risk.

Resumo Este estudo avaliou a soroprevalência de Toxoplasma gondii, Neospora caninum e Leptospira spp. em cães de Foz do Iguaçu, Paraná, Brasil. O teste de imunofluorescência indireta foi utilizado para detectar anticorpos anti-T. gondii e anti-N. caninum. Ensaio imunoenzimático e soroaglutinação microscópica foram utilizados para pesquisa de anticorpos anti-T. gondii e anti-Leptospira spp., respectivamente. Os resultados obtidos foram: 67,02% (435/649) das amostras reativas para T. gondii e, 1,38% (9/649) para N. caninum, ambas com ausência de variáveis significativas estatisticamente. Para Leptospira spp. os resultados indicaram soroprevalência de 23,11% (153/649). A análise das variáveis sem distinção de sorovar mostraram associação para caraterísticas intrísecas como raça, idade, estado nutricional e categoria de cães. Para as variáveis extrínsecas, a região da cidade e ter acesso à rua mostraram associação estatística (p<0,05). Os sorovares mais prevalentes foram: Canicola com 59,47% (91/153); Bratislava 13,07% (20/153) e Butembo 15,68% (24/153). As variáveis que compõem o modelo de análise multivariada ajustada usando como desfecho Leptospira spp. foram: idade, raça, estado nutricional e área. Para o sorovar Canicola, as variáveis significantes (p<0,05) foram sexo, estado nutricional e área; para o sorovar Bratislava, as variáveis significantes (p<0,05) foram linfadenomegalia e presença de pulgas. Dos resultados obtidos, cães podem ser usados como sentinelas para infecção por T. gondii e Leptospira spp. na cidade de Foz do Iguaçu e em outras cidades com desfechos similares. Além disso, medidas preventivas devem ser tomadas pelas autoridades de saúde, pois são zoonoses e os seres humanos também estão em risco.

Seroprevalencia de Bordetella pertussis en el personal de enfermería y médicos residentes del Hospital Infantil de México Federico Gómez / Seroprevalence of Bordetella pertussis in pediatric healthcare workers at the Hospital Infantil de México Federico Gómez

Resumen Introducción Bordetella pertussis es el agente causal de la tosferina, una enfermedad de alta letalidad, especialmente en menores de 6 meses, pero prevenible mediante la vacunación. Los reportes en hospitales de brotes de tosferina muestran que el caso índice suelen ser personas adultas. En adultos, la enfermedad se manifiesta principalmente con tos persistente. El propósito de este estudio fue conocer la seroprevalencia de B. pertussis en el personal de salud de un hospital pediátrico en un país donde aún no se considera la vacunación obligatoria para los empleados. Métodos Participaron personal de enfermería y médicos residentes en trato directo con pacientes hospitalizados. A cada participante se le realizó detección de anticuerpos inmunoglobulina G, antitoxina de pertussis (anti-TP) y se le aplicó un cuestionario para datos clínicos y demográficos. Resultados Se incluyeron 93 individuos, el 85% de personal de enfermería con mediana de edad de 35 años (rango intercuartil: 29-42.5). El 21.5% de los participantes laboraban en el Servicio de Urgencias, el 8.6%, en la Unidad de Terapia Intensiva Pediátrica, el 6.5%, en la Unidad de Cuidados Intensivos Neonatales. Se encontraron títulos detectables de anticuerpos anti-TP en el 18.3%, de los cuales, el 53% presentaron títulos de infección reciente y solamente el 23.5%, historia de tos de más de dos semanas de evolución. Conclusiones El personal de salud está en riesgo de sufrir la enfermedad y de transmitirla a los lactantes, quienes pueden fallecer por esta causa. Este estudio sugiere que las políticas actuales de vacunación en personal de salud se deben de modificar para determinar obligatoriedad de la vacuna, especialmente en quienes atienden a la población pediátrica.

Abstract Background Bordetella pertussis is the causative agent of pertussis, a disease that is preventable by vaccination but has a high mortality, particularly in children < 6 months. Reports of pertussis outbreaks in hospitals show that the index case is usually an adult. In adults, the disease manifests mainly with persistent cough. The purpose of this study was to determine the seroprevalence of B. pertussis in the health personnel of a pediatric hospital in a country where vaccination of this staff is not considered mandatory. Methods Nursing staff and resident doctors who were involved in direct treatment with hospitalized patients participated in the study. Each participant was screened for immunoglobulin G anti-pertussis toxin antibodies (anti-PT), and a questionnaire was applied for clinical and demographic data. Results Ninety-three individuals were included, of which 85% were nurses, median age 35 years (interquartile range: 29-42.5). The participants worked in the emergency department (21.5%), in the Pediatric Intensive Care Unit (8.6%), and in the Neonatal Intensive Care Unit (6.5%). Detectable titers of anti-TP antibodies were found in 18.3%, of which 53% presented titles suggestive of recent infection and only 23.5% cough > 2 weeks of duration. Conclusions Health personnel are at risk of suffering from the disease and be potential transmitters to infants, who may die from this cause. This study suggests that the current vaccination policies in health personnel should be modified to determine the compulsory nature of the vaccination, especially in those individuals in charge of the care of the pediatric population.

Serorreacción y prevalencia de sífilis en donantes de un banco de sangre de Barranquilla, Colombia / Seroreactivity and prevalence of syphilis in donors at a blood bank in Barranquilla, Colombia

Resumen Introducción. La sífilis es una enfermedad de interés en salud pública por sus elevadas tasas de morbilidad y mortalidad. Objetivo. Determinar la serorreacción y la seroprevalencia de sífilis según las variables sociodemográficas de los donantes de un banco de sangre del distrito de Barranquilla, Colombia, durante 2015 y 2016. Materiales y métodos. Se hizo un estudio descriptivo de corte transversal basado en los resultados de las pruebas treponémicas y no treponémicas. Se analizaron las variables sociodemográficas de la población estudiada y se hizo un análisis univariado en el que se determinaron las frecuencias absoluta y relativa de cada una de las variables categóricas. Se determinó la serorreacción a Treponema pallidum y la prevalencia de la infección activa. Se utilizó la prueba de ji al cuadrado de Pearson para evaluar las diferencias entre las proporciones. Resultados. Se encontró una serorreacción de 1,86 % para la infección previa con T. pallidum y una prevalencia de 0,93 % para la infección activa, las cuales fueron más altas en hombres adultos y en adultos mayores, viudos, desempleados y personas residentes en otros municipios del departamento de Atlántico diferentes de Barranquilla y su área metropolitana. Se encontró una asociación significativa entre la sífilis y las variables de sexo y ocupación. Conclusión. Se registró una serorreacción elevada a T. pallidum en donantes de sangre, comparada con el promedio nacional. Se encontró asociación entre la sífilis, y las variables sociodemográficas de sexo y ocupación, principalmente.

Abstract Introduction: Syphilis is a public health concern given its high impact on morbidity and mortality. Objective: We aimed to determine the association of syphilis seroreactivity and seroprevalence with sociodemographic variables of donors at a blood bank in the district of Barranquilla, Colombia, during 2015 and 2016. Materials and methods: We conducted a descriptive cross-sectional study based on the results of the treponemal and nontreponemal tests and the sociodemographic variables of the study population. We performed a univariate analysis to determine the absolute and relative frequencies for each categorical variable. We determined the seroreactivity against Treponema pallidum and the prevalence of active syphilis infection, and we used Pearson's chi-square test to evaluate the differences between the proportions. Results: We found a seroreactivity of 1.86% in individuals with previous T. pallidum infection, and a prevalence of 0.93% in those with active T. pallidum infection. These values were higher in adult men and older adults, widowers, the unemployed, and people living in municipalities of the Department of Atlántico other than Barranquilla and its metropolitan area. The incidence of syphilis infection showed a significant association with sex and occupation. Conclusion: In comparison to the national average, syphilis seroreactivity was high among blood donors. There was an association between syphilis infection incidence and sociodemographic variables such as sex and occupation.

Incidencia y subregistro de casos de leptospirosis diagnosticados con tres métodos diferentes en Urabá, Colombia / Incidence and underreporting of leptospirosis comparing three diagnostic methods in the endemic region of Urabá, Colombia

Resumen Introducción. La leptospirosis representa un problema de salud pública y es una causa importante de morbimortalidad en la región de Urabá, cuya notificación se ve afectada por las deficiencias en el diagnóstico. Objetivo. Establecer la incidencia de la leptospirosis en los municipios del llamado 'eje bananero' de la región de Urabá, documentar la magnitud del subregistro y proponer orientaciones para el diagnóstico por laboratorio por parte de la red de salud pública. Materiales y métodos. Se compararon dos fuentes de información sobre la leptospirosis: el sistema oficial nacional de vigilancia y un estudio transversal de 479 pacientes febriles, llevado a cabo entre abril de 2010 y mayo de 2012. El diagnóstico se hizo con base en tres pruebas: inmunofluorescencia indirecta, microaglutinación y hemocultivo. La exhaustividad de cada fuente de información se estimó mediante el método de captura y recaptura. Resultados. El 58 % (278/479) de los pacientes fueron positivos para leptospirosis, por lo menos, en una de las pruebas y, el 10,43 % (29/278), en las tres. La inclusión de una cepa nativa en el panel de la prueba de microaglutinación aumentó el porcentaje de positividad en 15 %. La tasa acumulada de incidencia fue de 66,5 por 100.000 habitantes y la proporción de letalidad fue de 2,15 %. El subregistro de la morbilidad por leptospirosis en la región de Urabá, fue de 27,8 % y, el de la mortalidad, de 66,6 %. Conclusión. El subregistro de leptospirosis en la región reitera la necesidad de usar más de una prueba diagnóstica para identificar Leptospira spp. en pacientes de zonas endémicas. Este subregistro podría ser una situación común en todo el país.

Abstract Introduction: Leptospirosis represents a public health problem and is a significant cause of morbidity and mortality in the region of Urabá. However, its notification reveals diagnostic limitations. Objective: To establish the incidence of leptospirosis in the municipalities of the so-called eje bananero in the Urabá region, to describe the magnitude of underreporting, and to propose guidelines for laboratory diagnosis by the public health network. Materials and methods: Two leptospirosis information sources were used: The national official surveillance system and a cross-sectional study of 479 acute-phase patients from April, 2010, to May, 2012. The diagnosis was made using three different tests: Indirect immunofluorescence, microagglutination test, and blood cultures. The exhaustiveness percentage of each information source was calculatedusing thecapture and recapture test. Results: From the total number of cases, 58% (278/479) were positive for leptospirosis at least by a test and 10.43% (29/278) of cases were positive by all three methods. The inclusion of a native strain in the microagglutination test panel increased the percentage of positivity by 15%. The cumulative incidence rate was 66.5/100,000 inhabitants and the case fatality ratio was 2.15%. The underreporting rates of leptospirosis in the Urabá region were 27.8% in morbidity and 66.6% in mortality. Conclusion: Under-registration of leptospirosis in the region highlights the necessity to use more than one diagnostic test to identify Leptospira in patients from endemic areas. Under-registration could be a common situation throughout the country.

Ehrlichia spp. infection in domestic cats from Rio de Janeiro State, southeast Brazil / Infecção de Ehrlichia spp. em gatos domésticos da Grande Rio de Janeiro, sudeste do Brasil

Abstract Ehrlichiosis is caused by agents belonging to Ehrlichia genus. Despite the frequent reports on the serological and molecular detection of E. canis in dogs in Brazil, there is scant data on ehrlichiosis in brazilian cats. This study aimed at investigating the occurrence of Ehrlichia spp. in domestic cats from Greater Rio de Janeiro, and evaluating hematological changes associated with this rickettsial infection. We searched for IgG antibodies against E. canis on blood samples of 216 cats by Indirect Fluorescence Assay (IFA). Additionally, we performed nested PCR (nPCR) and real-time PCR (qPCR) assays targeting E. canis-16S rRNA and dsb gene, respectively. Fifty-seven (26.4%) cats were seropositive for Ehrlichia spp. by IFA. Ehrlichia spp.-16S rRNA gene fragments were detected in 3 cats (1.4%). Although the obtained 16S rRNA sequences showed 99 to 100% identity with E. canis, cats were negative in qPCR. Anemia, thrombocytopenia, leukocytosis, left shift neutrophil and hyperproteinemia were observed. Anemia was statistically associated with seropositivity to E. canis and kittens showed lower positivity rates (p<0.05). This study showed that Ehrlichia spp. occur in domestic cats from Greater Rio de Janeiro. Further studies involving culture isolation are much needed to more precisely characterize these organisms.

Resumo A erliquiose é causada por agentes pertencentes ao gênero Ehrlichia . Apesar dos frequentes relatos de detecção sorológica e molecular de E. canis em cães no Brasil, existem poucos dados sobre a erliquiose em gatos brasileiros. Este estudo teve como objetivo investigar a ocorrência de Ehrlichia spp. em gatos domésticos do Grande Rio de Janeiro e avaliar as alterações hematológicas associadas a essa infecção rickettsial. Procuramos anticorpos IgG anti-E. canis em amostras de sangue de 216 gatos por Reação de Imunofluorescência Indireta (RIFI). Além disso, foram realizados ensaios de nested PCR (nPCR) e PCR em tempo real (qPCR) para detecção dos genes E. canis-16S rRNA e dsb , respectivamente. Cinquenta e sete (26,4%) gatos foram soropositivos para Ehrlichia spp. pela RIFI. Fragmentos do gene rRNA de Ehrlichia spp.-16S foram detectados em 3 gatos (1,4%) por ensaios de nPCR. Embora as sequências 16S rRNA obtidas tenham 99 a 100% de identidade com E. canis, os gatos foram negativos nos ensaios de qPCR. Anemia, trombocitopenia, leucocitose, desvio nuclear neutrofílico à esquerda e hiperproteinemia foram observados. Anemia foi estatisticamente associada à soropositividade para E. canis e filhotes apresentaram menores taxas de positividade (p <0,05). Este estudo demonstra que Ehrlichia spp. ocorrem em gatos domésticos da Grande Rio de Janeiro. Outros estudos envolvendo o isolamento por cultura são necessários para caracterizar com mais precisão esses organismos.

Circulation of spotted fever group rickettsiae among dogs seropositive for Leishmania spp. in an urban area of Brazil

Abstract INTRODUCTION: Dogs play an epidemiological role in several vector-borne diseases that affect human and animal health worldwide. We aimed to identify rickettsial circulation among dogs with canine visceral leishmaniasis (CVL) from a region endemic for both diseases. METHODS: CVL-seropositive dogs were screened for spotted fever group rickettsiae using an indirect immunofluorescence assay. RESULTS: Among the CVL-positive dogs, anti-Rickettsia rickettsii antibodies were identified in one asymptomatic and one oligosymptomatic dog. CONCLUSIONS: This study shows low circulation of antibodies to R. rickettsii in CVL-seropositive dogs. It is recommended that surveillance studies in dogs should continue in order to monitor this scenario.

Serological diagnosis and risk factors for Coxiella burnetii in goats and sheep in a semi-arid region of Northeastern Brazil / Diagnóstico sorológico e fatores de risco para Coxiella burnetii em caprinos e ovinos da região semiárida do nordeste do Brasil

Abstract Coxiella burnetii is a zoonotic agent transmitted mainly by small ruminants. In Brazil the disease has been classified as a notifiable disease since 2013, when human cases were reported. This study aimed to identify risk factors associated with the presence of anti- Coxiella burnetii antibodies in goats and sheep in a semiarid region of Northeastern Brazil. Sera of 412 goats and 403 sheep from municipality of Petrolina, Pernambuco, were examined by the Indirect Fluorescent Antibody Test (IFAT) against antigens of C. burnetii. Information about management variables (independent variables) that could be associated with the presence of the microorganism (dependent variables) were obtained from the supervisor of each farm. It was determined that 2.2% (9/412) of the goats and 2.1% (9/403) of the sheep had antibodies reactive to C. burnetii. The presence of anti-C. burnetii antibodies was associated with the dry area of the Sequeiro (a region in the northern part of the municipality of Petrolina) (P = 0.025), male sheep (P = 0.020), and intensive goat breeding (P = 0.005). This study therefore showed the presence of anti-C. burnetii antibodies in goat and sheep, confirming for the first time that this agent is likely circulating among goat herds in the Caatinga Biome, semi-arid of Brazil.

Resumo Coxiella burnetii é um agente zoonótico transmitido principalmente por pequenos ruminantes. No Brasil, a doença foi classificada como de notificação compulsória desde 2013, quando casos humanos foram relatados. O objetivo deste estudo foi identificar os fatores de risco associados à presença de anticorpos anti-Coxiella burnetii em caprinos e ovinos em uma região semiárida do Nordeste do Brasil. Este estudo envolveu um inquérito sorológico de 412 caprinos e 403 ovinos em fazendas do município de Petrolina, no estado de Pernambuco. Os soros foram examinados pela Reação de Imunofluorescência Indireta (RIFI) contra antígenos de C. burnetii . Informações sobre variáveis ​​de manejo (variáveis ​​independentes) que poderiam estar associadas à presença do microrganismo (variáveis ​​dependentes) foram obtidas do proprietário de cada fazenda. Foi determinado que 2,2% (9/412) dos caprinos e 2,1% (9/403) dos ovinos tinham anticorpos reativos a C. burnetii. A presença de anticorpos anti-C. burnetii foram associados com a área seca do Sequeiro (região no norte do município de Petrolina) (P = 0,025), ovinos machos (P = 0,020) e criação intensiva de caprinos (P = 0,005). Este estudo, portanto, observou a presença de anticorpos anti-C. burnetii em pequenos ruminantes, confirmando pela primeira vez que este agente pode estar circulando em rebanhos caprinos no bioma Caatinga, semiárido do Brasil.

Occurrence of serological reactions for serogroup Sejroe (CTG and Prajtino) in female buffalo in the state of Pernambuco, Brazil

ABSTRACT The objective of this study was to evaluate the occurrence of anti-Leptospira spp. antibodies in female buffalo in the state of Pernambuco. A total of 123 female buffalo blood samples were collected from five properties distributed in the state of Pernambuco. The microscopic agglutination test was used to study anti-Leptospira spp. antibodies. The occurrence of anti-Leptospira spp. antibodies was 28.5% (35/123; CI 20.7-37.3%) and on different properties, the occurrence ranged from 28.6% to 80.0%, with 100% of the properties showing animals with positive results. The serovars of the serogroup Sejroe with a higher incidence were Hardjoprajtino (CTG strain, 49.1%) and Hardjo (Prajtino genotype, 43.2%), followed by serogroup Grippotyphosa with the Grippotyphosa serovar (3.9%), serogroup Pomona with the Pomona serovar (1.9%), and the Icterohaemorrhagiae serovar Copenhageni (1.9%). This was the first record of the occurrence of anti-Lepstospira spp. antibodies in female buffalo in the state of Pernambuco. Control measures are necessary to prevent health and economic losses, given that the agent involved affects animal reproduction, triggering drops in conception rates or even clinical cases of abortion.

Bartonella henselae: evidencia serológica en pacientes pediátricos con sospecha clínica de enfermedad por arañazo de gato / Bartonella henselae: Serological evidence in pediatric patients with clinical suspicion of cat scratch disease

La enfermedad por arañazo de gato (EAG) es producida por Bartonella henselae.Afecta principalmente a ninos y el reservorio es el gato doméstico. El diagnóstico de laboratorio se basa en la detección de anticuerpos por inmunofluorescencia indirecta (IFI). El objetivo de este trabajo fue analizar la evidencia serológica de infección por B. henselae en pacientes pediátricos que reunían criterios clínicos/epidemiológicos para la sospecha de EAG. Se estudió a 92 pacientes; de acuerdo con los resultados serológicos, estos fueron categorizados en 4 grupos: 1) IgG (+)/IgM (+), 31,5% (n = 29);2) IgG (-)/IgM (+), 10,9% (n = 10);3) IgG (+)/IgM (-), 9,8% (n = 9), y 4) IgG (-)/IgM (-), 47,8% (n = 44). La divulgación de estos resultados intenta promover futuros trabajos que investiguen la seroprevalencia de Bartonella spp. en Argentina. Esto permitirá conocer la importancia de esta zoonosis en nuestra población y evaluar nuevos puntos de corte para esta técnica serológica.

Cat scratch disease (CSD) is caused by Bartonella henselae, which mainly affects children. The cat is the reservoir. The laboratory diagnosis is based on the detection of antibodies by the Indirect Immunofluorescence (IFI) assay. The objective of this study was to analyze the serological evidence of B. henselae infection in pediatric patients that met the clini-cal/epidemiological criteria for suspected CSD. We studied 92 patients, who were categorized into four serological groups: 1) IgG (+)/IgM(+), 31,5% (n = 29); 2) IgG (-)/IgM(+), 10,9% (n = 10); 3) IgG (+)/IgM(-), 9,8% (n = 9); 4) IgG (-)/IgM(-), 47,8% (n = 44). These findings aim to promote future works for investigating the seroprevalence of Bartonella spp. in Argentina, which will allow us to know the importance of this zoonosis in our population and to evaluate new cut-off points of the technique.

Development and validation of a whole-cell ELISA for serologically diagnosing Helicobacter pylori infection in Brazilian children and adults: a diagnostic accuracy study

ABSTRACT BACKGROUND: Serological tests are practical, with low cost, but no noninvasive tests are available for diagnosing Helicobacter pylori (H. pylori) infection in Brazil. The aim here was to develop and validate enzyme-linked immunosorbent assay (ELISA) serological tests to detect anti-H. pylori immunoglobulin G antibodies, based on cultured strains from Brazilian patients. DESIGN AND SETTING: Cross-sectional, diagnostic accuracy study comparing a locally developed and validated ELISA and invasive tests among dyspeptic patients at two public hospitals in São Paulo, Brazil. METHODS: An ELISA test was prepared using whole-cell antigen from 56 strains. After genotypic characterization, it was standardized and optical density (OD) cutoffs were determined based on the serum antibody response of 100 H. pylori-negative samples, compared with 82 H. pylori-positive samples. Validation was performed on 174 symptomatic patients. RESULTS: The optimal OD cutoffs established (for monoclonal and polyclonal tests, respectively) were 0.167 and 0.164; overall ELISA sensitivity: 84.3%, 78.9%; specificity: 88.6%, 90.6%; positive predictive value (PPV): 75.4%, 80%; negative predictive value (NPV): 93.1%, 81.8%; accuracy: 87.3%, 86.2%; child and adolescent ELISA sensitivity: 74.2%, 81.8%; specificity: 90.8%, 86.7%; PPV: 66.6%, 84.3%; NPV: 95.8%, 84.8%; accuracy: 88.5%, 84.6; adult ELISA sensitivity: 84.4%, 75%; specificity: 86.9%, 93%; PPV: 81.8%, 78.3%; NPV: 88.9%, 91.8%; accuracy: 85.9%, 88.5%. CONCLUSION: The polyclonal serological test developed using local strains presented better diagnostic performance among children and adolescents, while the monoclonal test was better among adults. The results from both tests suggest that these in-house serological tests could be used to detect anti-H. pylori antibodies in our population, for screening purposes.

Evaluation of dot-blot test for serological diagnosis of bovine brucellosis

Abstract The objective of this study was to standardize and validate the dot-blot test for the serological diagnosis of bovine brucellosis, compare the results with those found in the 2-mercaptoethanol (2-ME) and complement fixation test (CF), and estimate the relative sensitivity and specificity of the dot-blot compared to these tests. Fifty bovine blood serum samples were used for the test standardization, and 1315 samples were used for evaluation and comparison between the tests; the results were compared using the Kappa indicator. At the end of standardization, it was established as optimal for the antigen obtained from Brucella abortus B19 after passing through a microorganism rupture process, the blood serum samples diluted at 1:100, and the conjugate at 1:30,000. The comparison of the dot-blot results with 2-ME showed Kappa index of 0.9939, sensitivity of 99.48%, and specificity 99.91%, with CF, Kappa index of 0.8226, sensitivity 100% and specificity 95.32%. Using the combination of the test results 2-ME and CF to establish the true condition of the animal, the dot-blot showed relative sensitivity of 100%, and relative specificity of 99.91%. The evaluated test proved to be effective and reliable, besides being easy to handle and interpret the results.

Detection of antibodies against Leptospira spp in free-living marsupials caught in the Eastern Amazon

Abstract INTRODUCTION: Serological surveys are important to assess the health status of wild animals. In this study, antibodies against Leptospira spp, causal agents of leptospirosis, were detected in free-living marsupials in the State of Pará, Brazil. METHODS: Nineteen blood samples collected from marsupials in the municipalities of Peixe-Boi, Viseu, and Castanhal were subjected to microscopic agglutination tests. RESULTS: In total, 36.8% (7/19) of samples were positive, and two exhibited co-agglutination. The most frequent serovars were Icterohaemorrhagiae (60%; 3/5), Panama (20%; 1/5), and Nupezo (20%; 1/5). CONCLUSIONS: Anti-Leptospira spp antibodies currently circulate in free-living marsupials in Northeastern Pará.