Hemorragia alveolar pulmonar como presentación poco frecuente de infección por Leptospira spp: una enfermedad ocupacional del personal militar en zonas tropicales / Pulmonary alveolar hemorrhage as an unusual presentation of Leptospira spp: an occupational-disease of military personnel in tropical zones

Resumen La leptospirosis es una de las zoonosis endémicas más importantes en el mundo con un aumento de la incidencia en los últimos años. En el personal militar podría ser catalogada como una enfermedad ocupacional dado sus actividades específicas en áreas rurales. Su presentación clínica es variable siendo en la mayoría de los casos una enfermedad febril autolimitada. De acuerdo con diversos factores dependientes del patógeno y del hospedero pueden presentarse manifestaciones severas de la enfermedad dentro de la cual destaca el compromiso pulmonar con una alta tasa de mortalidad. Existe evidencia del uso de esteroide sistémico como parte del tratamiento de esta complicación. Presentamos el caso de un paciente joven, militar, que debuta con síndrome de hemorragia alveolar difusa secundario a leptospirosis y presenta una excelente respuesta al tratamiento con altas dosis de metilprednisolona, con una discusión del proceso diagnóstico y aspectos fisiopatológicos de esta condición.

Leptospirosis is one of the most important endemic zoonoses in the world with an increase in incidence in recent years. In military personnel it could be classified as an occupational disease given their specific activities in rural areas. Its clinical presentation is variable being in most cases a self-limited febrile disease. According to various factors dependent on the pathogen and the host, severe manifestations of the disease may occur within which the pulmonary involvement with a high mortality rate stands out. There is evidence of systemic steroid use as part of the treatment of this complication. We present a case of a young, military patient who debuts with diffuse alveolar hemorrhage syndrome secondary to leptospirosis and presents an excellent response to treatment with high doses of methylprednisolone, with a discussion of the diagnostic process and pathophysiological aspects of this condition.

Leptospirose: Características da enfermidade em humanos e principais técnicas de diagnóstico laboratorial / Leptospirosis: Characteristics of the human disease and main laboratory diagnostic techniques

A leptospirose é a zoonose de maior distribuição geográfica, com estimativa de cerca de 60.000 mortes por ano. A doença é causada por bactérias do gênero Leptospira, que possui mais de 300 diferentes sorovares e 64 espécies já identificadas, sendo o ambiente a principal fonte de contaminação. A doença em humanos apresenta manifestações clínicas variadas e caráter bifásico, devendo ser confirmada por meio do diagnóstico laboratorial. O objetivo deste trabalho foi reunir conceitos atualizados sobre a leptospirose humana e as principais técnicas de diagnóstico laboratorial empregadas. A MAT é considerada o padrão-ouro para o diagnóstico da leptospirose, mas devido à baixa sensibilidade na fase inicial da doença é necessário o emprego de técnicas mais sensíveis neste período. Baseado em diversos estudos, as metodologias de PCR, ELISA-IgM e teste rápido apresentaram sensibilidade satisfatória nos primeiros dias após o início dos sintomas. Na segunda semana, a MAT apresentou 100% de sensibilidade, mantendo sua alta especificidade em ambas as fases. No geral, os testes sorológicos de ELISA-IgM e teste rápido apresentaram resultados satisfatórios como métodos de diagnóstico precoce, principalmente tratando-se de locais com pouca infraestrutura, diferente dos laboratórios de referência onde é possível empregar as técnicas de PCR e MAT.

Leptospirosis is the most widespread zoonosis, which has a balance of almost 60,000 deaths per year. Bacteria of Leptospira genus, which has more than 300 different serovars and 64 species already identified, cause the disease, being the environment the main source of contamination. The human disease presents a large set of clinical manifestations, showing biphasic presentation, the reason why leptospirosis must be confirmed by laboratory diagnosis. This study aimed to group recent concepts concerning human leptospirosis and the main diagnosis techniques employed at the laboratory. MAT is considered the gold standard for leptospirosis diagnosis, but has low sensitivity on the onset of disease, leading to the use of techniques with higher sensitivity on this period. Based on several studies, PCR, ELISA-IgM and rapid test presented satisfactory sensitivity on the onset of symptoms. In the second week, MAT showed 100% of sensitivity, maintaining its high specificity in both phases. In general, the ELISA-IgM and rapid serological tests showed satisfactory results as methods for early diagnosis, especially in the case of places with poor infrastructure, different from the reference laboratories where it is possible to use the PCR and MAT techniques.

Prediction model to discriminate leptospirosis from hantavirus

SUMMARY OBJECTIVE: The aim of this study was to build a prediction model to discriminate precociously hantavirus infection from leptospirosis, identifying the conditions and risk factors associated with these diseases. METHODS: A logistic regression model in which the response variable was the presence of hantavirus or leptospirosis was adjusted. RESULTS: As a result, the method selected the following variables that influenced the prediction formula: sociodemographic variables, clinical manifestations, and exposure to environmental risks. All variables considered in the model presented statistical significance with a p<0.05 value. The accuracy of the model to differentiate hantavirus from leptospirosis was 88.7%. CONCLUSIONS: Concluding that the development of statistical tools with high potential to predict the disease, and thus differentiate them precociously, can reduce hospital costs, speed up the patient's care, reduce morbidity and mortality, and assist health professionals and public managers in decision-making.

Hematúria macroscópica em equinos associada à infecção por Leptospira interrogans / Macroscopic haematuria in horses associated with Leptospira interrogans infection

Hematúria é uma grave manifestação clínica de doença do sistema urinário, ocorrendo sob as formas micro ou macroscópica. Neste artigo relatam-se dois casos de hematúria macroscópica associada à infecção por Leptospira interrogans sorogrupo Canicola. O exame clínico inicial revelou hematúria macroscópica, taquicardia, taquipneia, febre, elevação do tempo de perfusão capilar, hipomotilidade intestinal, além de icterícia da mucosa oral. Leucocitose, proteinúria, glicosúria, piúria e azotemia foram achados comuns aos dois casos. Teste de Soroaglutinação Microscópica foi realizado para titulação de anticorpos contra Leptospira interrogans. Tratamento incluiu medidas terapêuticas de suporte (fluidoterapia), controle da hematúria e antibioticoterapia. Sete dias após manifestação dos sinais clínicos iniciais, ambos animais receberam alta hospitalar após remissão dos sinais clínicos.

Haematuria is a serious clinical manifestation of urinary system disease, occurring in micro or macroscopic forms. In this article two cases of macroscopic haematuria associated with Leptospira interrogans serogroup Canicolainfection are related. The initial clinical examination revealed macroscopic haematuria, tachycardia, tachypnea, fever, increased capillary perfusion time, intestinal hypomotility, in addition to jaundice of the oral mucosa. Leukocytosis, proteinuria, glycosuria, pyuria and azotemia were common findings in both cases. Microscopic serum agglutination test was performed for titration of antibodies against Leptospira interrogans. Treatment included supportive therapeutic measures (fluid therapy), hematuria control and antibiotic therapy. Seven days after the manifestation of the initial clinical signs, both animals were discharged from the hospital without complications.

Leptospirosis y rickettsiosis, reto diagnóstico para el síndrome febril en zonas endémicas / Leptospirosis and rickettsiosis, a diagnostic challenge for febrile syndrome in endemic areas

Resumen. Se presenta el caso de un hombre de 50 años de edad proveniente de la región de Urabá, Colombia, con una infección mixta por Rickettsia rickettsii y Leptospira interrogans serovar Copenhageni ST78, y pruebas negativas para malaria y dengue. El paciente presentó un síndrome febril que no mejoró con el tratamiento antibiótico sistémico y, finalmente, falleció en la unidad de cuidados intensivos. El diagnóstico post mortem se hizo mediante tipificación molecular de los dos agentes etiológicos. En la inspección del domicilio del paciente, se encontró un ejemplar de Rattus rattus infectado con L. interrogans del mismo serovar detectado en él. No se encontraron garrapatas en los animales domésticos que habitaban con el paciente. Se reporta una infección mixta con síntomas clínicos progresivos y fatales en un paciente con antecedentes laborales de riesgo en una zona endémica para enfermedades tropicales, lo que obliga a tener presente la posibilidad de infecciones simultáneas en personas procedentes de áreas endémicas que consulten reiteradamente por síndrome febril sin resolución y tengan riesgo laboral relacionado con actividades agrícolas.

Abstract. This is the case of a 50-year-old male from the region of Urabá, Colombia, with a mixed infection by Rickettsia rickettsii and Leptospira interrogans serovar Copenhageni ST78 and negative test for malaria and dengue fever. The patient presented with febrile syndrome and was unresponsive to systemic antibiotic treatment, who finally died in the intensive care unit. We established the postmortem diagnosis through molecular typification of the two etiological agents. In the inspection at the patient's home, we found a Rattus rattus specimen infected with L. interrogans of the same serovar found in him. We found no ticks parasitizing the domestic animals cohabitating with the patient. This case of a mixed infection with progressive and fatal symptoms in a patient with occupational risk in a tropical disease endemic zone highlights the importance of considering the potential presentation of simultaneous etiologies in patients with multiple medical visits for unresolved febrile syndromes associated with risky exposure during agricultural activities.

Leptospirosis, una zoonosis que impacta a la salud: diagnóstico, tratamiento y nuevas alternativas de control / Leptospirosis a zoonosis that impacts health: diagnosis, treatment and new alternatives of control

La leptospirosis es una zoonosis con potencial epidémico y de difícil diagnóstico que requiere un manejo integral para orientar las medidas de prevención y control; sin embargo, una de las dificultades es la existencia de más de 300 serovares, la supervivencia de la bacteria en el ambiente por más de 180 días y la importancia del agua como vehículo de transmisión. Esto asociado con los efectos adversos de los antibióticos y su efecto sobre la multirresistencia generada por la mayoría de las bacterias, hace que se evalúen nuevas alternativas a partir de la biodiversidad. Por lo tanto, el objetivo de este artículo es abordar la leptospirosis y su diagnóstico enfatizando en el control convencional de la infección y las alternativas de tratamiento a partir del uso de plantas medicinales. Para esto se realizó una revisión exhaustiva de artículos en bases de datos. La información encontrada permitió establecer los aspectos relevantes de la enfermedad, su diagnóstico y tratamiento, tanto con antimicrobianos convencionales como frente a nuevas alternativas de origen natural. Se concluye que es importante realizar investigaciones orientadas hacia la búsqueda de principios activos que puedan contribuir al control de Leptospira spp., agente causal de la leptospirosis, una de las zoonosis más importantes por su impacto en salud humana, veterinaria y del ecosistema(AU)

Leptospirosis is a potentially epidemic zoonosis of difficult diagnosis which requires comprehensive management to indicate appropriate prevention and control measures. However, some of the difficulties are the existence of more than 300 serovars, survival of the bacteria in the environment for more than 180 days, and the role of water as a route of transmission. The above situation, alongside the adverse effects of antibiotics and their effect on the multi-drug resistance developed by most bacteria, lead to the search for new alternatives based on biodiversity. The purpose of the study was therefore to address leptospirosis and its diagnosis highlighting conventional control of the infection as well as treatment options based on the use of medicinal plants. To achieve this end, an exhaustive review was conducted of papers included in databases. The information obtained made it possible to determine the relevant aspects of the disease, its diagnosis and its treatment with conventional antimicrobials as well as new alternatives of a natural origin. Conclusions point to the importance of conducting research aimed at the search for active principles potentially contributing to control of Leptospira spp., the causative agent of leptospirosis, one of the most relevant zoonoses in terms of its impact on the health of humans, animals and the ecosystem(AU)

Molecular biology as a diagnostic tool for detection of Leptospira spp. in cows of a border region ­ case report

The reproductive efficiency of livestock is the basis for the success of livestock, dairy or beef, and having high reproductive performance depends on several factors within the production system and the presence of infectious diseases of the reproductive sphere in the herd is one of the factors that can compromise that efficiency. The aim of this study was to use molecular biology as a diagnostic tool for the detection of Leptospira spp. DNA in cows with reproductive disorders on a rural property in the municipality of Boca do Acre, Amazonas, Brazil. Vaginal mucus was collected from nine Nelore breeding cows with a history of abortion and birth of weak calves submitted to DNA extraction and nested-PCR technique for 16S gene amplification at the bacterial genus level. Of the nine samples analyzed, five (55.55%) amplified a product of 331bp. The municipality of Boca do Acre is bordered by Peru and Bolivia, and knowledge of the prevalence of the disease, serovars, and circulating Leptospira species is essential for the adoption of measures related to animal husbandry, as well as health education for ranchers and their workers to avoid a possible occupational infection since this disease is considered an important zoonosis. New molecular studies using primers that allow the identification of the Leptospira species and mainly pathogenic species should be conducted in this region in order to elucidate the possible species of this etiological agent and the possible reservoirs of the disease to begin the understanding of the epidemiology of this disease in cattle in this region of border.

Serological survey of leptospirosis, brucellosis, and lentivirus in herds of small ruminants in Recôncavo Baiano, Bahia, Brazil / Inquérito sorológico de leptospirose, brucelose e lentiviroses em rebanhos de pequenos ruminantes no Recôncavo Baiano, Bahia, Brasil

A serological survey was carried out to assess the frequency of leptospirosis, small ruminants lentivirus (SRLV), and brucellosis in small ruminant herds in the Recôncavo Baiano, State of Bahia, Brazil, from February to December 2017. In four goat herds, 125 animals were tested for SRLV and leptospirosis, while in five sheep herds, 378 animals were tested for leptospirosis, brucellosis, and SRLV. Regarding leptospirosis, MAT detected 80% of goats and 15.34% of sheep seroreactive. Reactivity was most frequent to serogroups Autumnalis and Grippotyphosa in goats and sheep, respectively. Regarding SRLV, 8.8% of goats and 0.79% of sheep were reactive. Search for anti-B. ovis antibodies revealed 0.52% reactivity. In sheep, three animals showed simultaneous seroreactivity for SRLV and leptospirosis, while one animal had a serological response for brucellosis and leptospirosis. In goats, simultaneous seroreactivity for SRLV and leptospirosis was observed in only one animal. Leptospirosis was the most frequent of the three infectious diseases in investigated herds.(AU)

Foi realizado um inquérito sorológico para avaliar a frequência de ocorrência de leptospirose, lentiviroses de pequenos ruminantes (LVPR) e brucelose em rebanhos de pequenos ruminantes no Recôncavo Baiano, estado da Bahia, Brasil, no período de fevereiro a dezembro de 2017. Em quatro rebanhos de caprinos, foram testados 125 animais para LVPR e leptospirose, enquanto em cinco rebanhos de ovinos, foram testados 378 animais para leptospirose, brucelose e LVPR. Em relação à leptospirose, 80% das cabras e 15,34% das ovelhas foram sororreativas. Os sorogrupos de Leptospira spp. predominantes foram Autumnalis e Grippotyphosa para caprinos e ovinos, respectivamente. Em relação as LVPR, 8,8% dos caprinos e 0,79% dos ovinos foram reativos. Adicionalmente, a pesquisa de anticorpos Anti-B. ovis revelou 0,52% de ovinos reativos. Em ovinos, três animais apresentaram sororreatividade simultânea para LVPR e leptospirose, enquanto um animal teve resposta sorológica para brucelose e leptospirose. Em caprinos, sororreatividade simultânea para LVPR e leptospirose foi observada em apenas um animal. A leptospirose foi a doença infecciosa mais frequente nos rebanhos investigados.(AU)

Occurrence of Leptospira spp. and factors associated with the infection in horses from a military contingent in the Rio Grande do Sul State, Brazil / Ocorrência de Leptospira spp. e fatores associados à infecção em equinos de contingente militar no estado do Rio Grande do Sul, Brasil

The seroprevalence of Leptospira spp. and infection risk factors were investigated in horses from a military contingent in the Rio Grande do Sul State, Brazil. A total of 446 horses were evaluated and categorized into 309 mares, 11 stallions, and 126 horses with an average age of three years. To determine seroprevalence, serum samples from all horses were submitted to the microscopic agglutination test against 12 serovars belonging to nine serogroups, usually circulating in equine populations. To investigate the possible risk factors of infection by Leptospira spp., questionnaires were applied in that military unit. The seroprevalence in the horses with an average age of 3 years, mares, and stallions were 57.94% (73/126), 54.05% (165/309), and 45.45% (5/11), respectively. The main risk factors identified were contact with different water sources, the presence of rodents, and contact with wild and domestic animals. The high seroprevalence of anti­Leptospira spp. antibodies may be associated with the presence of these risk factors. Therefore, the exposure of horses to possible reservoirs of Leptospira spp. should be minimized. Also, the immunoprophylaxis protocol should be reviewed and a shorter interval between vaccinations adopted to control leptospirosis in this herd.(AU)

A soroprevalência de Leptospira spp. e os fatores de risco para infecção foram investigados em cavalos de um contingente militar no Rio Grande do Sul, Brasil. Um total de 446 cavalos foram avaliados e categorizados em 309 éguas, 11 garanhões e 126 cavalos com idade média de três anos. Para determinação da soroprevalência, amostras de soro de todos os equinos foram submetidas ao teste de aglutinação microscópica contra 12 sorovares pertencentes a nove sorogrupos, normalmente circulantes em populações equinas. Para investigar os possíveis fatores de risco da infecção por Leptospira spp., foi aplicado um questionário naquela unidade militar. A soroprevalência nos cavalos com idade média de três anos, éguas e garanhões foi de 57,94% (73/126), 54,05% (165/309) e 45,45% (5/11), respectivamente. Os principais fatores de risco identificados foram o contato com diferentes fontes de água, a presença de roedores e o contato com animais silvestres e domésticos. A alta soroprevalência de anticorpos anti­Leptospira spp. pode estar associada à presença desses fatores de risco. Portanto, a exposição destes equinos aos possíveis reservatórios de Leptospira spp. deve ser minimizada. Além disso, o protocolo de imunoprofilaxia deve ser revisto e, possivelmente, um menor intervalo entre as vacinas deve ser adotado para o controle da leptospirose neste plantel.(AU)

Leptospirosis diagnosis among patients suspected of dengue fever in Brazil

The early symptoms of leptospirosis and dengue fever are difficult to distinguish and can cause diagnostic confusion. Due to the large dengue epidemics that has occurred in Brazil in recent years, it is possible that cases of leptospirosis were unreported. Therefore, we performed a retrospective study to detect leptospirosis in patients who were tested for dengue, but whose laboratory diagnoses were negative. Methods: Sera samples from 2,017 patients from 48 cities located in the central region of São Paulo state, Brazil, were studied. All samples were subjected to the microscopic agglutination test (MAT), 305 of which were taken from patients five days or less since the onset of symptoms, and were additionally subjected to real-time polymerase chain reaction (PCR). Results: The overall prevalence of leptospirosis cases was 21 (1.04%), with 20 through MAT (18 for Icterohaemorrhagiae and two for the Cynopteri serogroup) and one through PCR (amplicon sequencing compatible with Leptospira interrogans). According to previously established criteria, eight cases of leptospirosis were classified as "confirmed" and 13 as "probable". The Brazilian notification system for health surveillance had no records for 16 patients positive for leptospirosis and, thus, they were considered unreported cases. Statistical analyses revealed that the prevalence of leptospirosis was higher in men (1.56%) than in women (0.56%), and the mean age was higher in positive patients (43.7 years) than in negative ones (32.3 years). Conclusion: The results indicated that patients suspected of dengue fever had evidence of leptospirosis or Leptospira infection, and most of these cases were unreported in the Brazilian notification system. The high burden of dengue may contribute to the misdiagnosis of leptospirosis, and health professionals should increase their awareness of leptospirosis as an important differential diagnosis of patients with suspicion of dengue.(AU)

Seroprevalencia de anticuerpos contra Leptospira spp. en cerdos criados en Portoviejo, Ecuador / Seroprevalence of antibodies against Leptospira spp. in pigs raised in Portoviejo, Ecuador

Introducción: La leptospirosis es una enfermedad infecciosa bacteriana causada por leptospiras patógenas, que afectan a una amplia variedad de mamíferos incluido el hombre. Es considerada como una de las enfermedades zoonóticas de mayor distribución a nivel mundial. Objetivo: Determinar la seroprevalencia de anticuerpos contra Leptospira spp. y factores de riesgo asociados a la leptospirosis en algunas poblaciones de cerdos de Portoviejo. Métodos: Estudio descriptivo, realizado entre octubre de 2018 a marzo de 2019. Se estudiaron cerdos criados en parroquias del cantón Portoviejo, Manabí, Ecuador. Se colectaron un total de 280 muestras de sangre (115 de cerdos en crianza tecnificada y 165 en crianza de traspatio), 140 en el periodo lluvioso (enero-marzo) y 140 en el periodo seco (octubre-diciembre), de forma tal que fueran representativos los animales para ambas épocas. El diagnóstico serológico se realizó mediante la prueba de aglutinación microscópica. Resultados: En la crianza tecnificada, la seroprevalencia fue de 16,52 por ciento (19/115) y en la crianza de traspatio, 20,61 por ciento (34/165). Se presentó con mayor frecuencia en serovares Australis e Icterohaemorrhagiae en 14,28 por ciento (40/280), respectivamente, y Bataviae en 13,21 por ciento (37/280). El riesgo asociado a la infección por leptospiras fue mayor en los cerdos del sexo femenino, en las poblaciones de cerdos que consumieron agua de ríos y en los predios donde no se realizó un control de roedores silvestres. Conclusiones: La seroprevalencia a anticuerpos contra Leptospira spp. en la población de cerdos en crianza tecnificada y traspatio fue moderada, pero constituye un peligro potencial para la población humana expuesta. En ambos sistemas de crianza, los serovares más frecuentes fueron Australis, Icterohaemorrhagiae y Bataviae(AU)

Introduction: Leptospirosis is an infectious disease caused by pathogenic Leptospira sp. bacteria. It affects a large number of mammals, including humans, and is considered to be one of the most widely distributed zoonotic diseases worldwide. Objective: Determine the seroprevalence of antibodies against Leptospira spp. and the risk factors associated to leptospirosis in some pig populations from Portoviejo. Methods: A descriptive study was conducted from October 2018 to March 2019 of pigs raised in parishes from Portoviejo Canton in Manabí, Ecuador. A total 280 blood samples were collected (115 from pigs raised technically and 165 from pigs raised in backyards), 140 in the rainy season (January-March) and 140 in the dry season (October-December), so that the samples were representative of the two seasons. Serological diagnosis was performed with the microscopic agglutination test. Results: In technical breeding, seroprevalence was 16.52 percent (19/115), whereas in backyard breeding it was 20.61 percent (34/165). It was most common in serovars Australis and Icterohaemorrhagiae, with 14.28 percent (40/280), respectively, and Bataviae with 13.21 percent (37/280). Risk associated to Leptospira infection was greater in sows, in pig populations which drank river water, and in areas where wild rodent control was not present. Conclusions: Seroprevalence of antibodies against Leptospira spp. was moderate in technical and backyard pig raising populations, but it is a potential hazard for the human population exposed. In both breeding systems, the most common serovars were Australis, Icterohaemorrhagiae and Bataviae(AU)

Leptospirosis en pediatría, un diagnóstico a tener en cuenta / Leptospirosis in pediatrics, a diagnosis to be considered

Resumen La leptospirosis humana es una importante causa de enfermedad febril aguda que afecta con mayor frecuencia las zonas tropicales. Su principal reservorio es los roedores por lo que la infección está vinculada con factores de pobreza, déficit sanitario, actividades recreativas y laborales que favorecen el contacto con la orina de animales infectados, agua o suelos contaminados. Las infecciones por Leptospira spp tienen una presentación inespecífica desde cuadros asintomáticos hasta un compromiso multisistémico con alta mortalidad, siendo la sospecha clínica y la presencia de factores de riesgo las variables iniciales para el diagnóstico de la enfermedad. Entre los métodos disponibles para el diagnóstico microbiológico se destaca la observación directa, el cultivo, la reacción de polimerasa en cadena y la serología que permiten la confirmación de éste. El tratamiento de leptospirosis en niños se basa en medidas de soporte y en el inicio de antimicrobianos β-lactámicos o macrólidos. Las medidas de prevención de la enfermedad son fundamentales en la población expuesta al riesgo siendo recursos esenciales intervenciones generales, quimioprofilaxis y vacunación. El conocimiento, la consideración diagnóstica y el tratamiento oportuno en el paciente pediátrico, acelera la recuperación y limita la aparición de complicaciones que pueden impactar en la calidad de vida.

Abstract Human leptospirosis is an important cause of acute febrile disease that most frequently affects tropical areas. Its main reserve is rodents, so the infection is limited by factors of poverty, health deficit, recreational and work activities that favor contact with the urine of infected animals, contaminated water or soil. Leptospira spp infections have a non-specific presentation, ranging from asymptomatic to multisystem compromise with high mortality; being the clinical suspicion and the presence of risk factors the initial variables for the diagnosis of the disease. Among the available methods for microbiological diagnosis, direct observation, culture, polymerase chain reaction and serology that allows confirmation of this are highlighted. The treatment of leptospirosis in children is based on clinical support measures and the antibiotic initiation of β-lactams or macrolides. Disease prevention measures are fundamental in the population exposed to risk being essential resources general interventions, chemoprophylaxis and vaccination. The knowledge, diagnostic consideration, and timely treatment in pediatric patients accelerate recovery and limit the appearance of complications that can impact the quality of life

Microscopio agglutination test: Variables that affect the time of serological confirmation of human leptospirosis cases / Test de microaglutinación: variables que afectan el tiempo de confirmación de los casos humanos de leptospirosis

Abstrac Leptospirosis is considered an endemic disease in Buenos Aires province, Argentina,with human cases reported annually from rural and urban areas. The aim of the study was todescribe the variables that influence the delay in the serological confirmation of leptospirosisin human cases (period 2006-2014) from Buenos Aires province. Sixty-four percent (64%) ofcases could be confirmed by microscopic agglutination test (MAT) with the first sample. The time of confirmation of the human leptospirosis cases was on average 21 days from the onset of the first clinical signs, and varied depending on the distance of the different sanitary regions. The geographical distribution of the confirmed leptospirosis human cases, in addition to the high number of suspected cases and probable cases (which could not be confirmed by MAT), demonstrate that leptospirosis is endemic and underreported in Buenos Aires province, and that distance and lack of resources could be determinant factors of this situation.

Resumen La leptospirosis se considera una enfermedad endémica en la provincia de Buenos Aires, con casos humanos registrados anualmente en áreas rurales y urbanas. El objetivo de este estudio fue describir las variables que influyen en el retraso de la confirmación serológica de la leptospirosis en humanos, a partir del análisis de los casos ocurridos en el período 2006-2014 en la provincia de Buenos Aires. El 64% de los casos fue confirmado por microaglutinación (MAT) con la primera muestra clínica. El tiempo de confirmación promedio fue de 21 días desde el inicio de los primeros signos clínicos; este valor se vio afectado por la distancia entre los centros de salud y los laboratorios de referencia en las distintas regiones sanitarias. La distribución geográfica de los casos confirmados de leptospirosis en humanos, además del alto número de casos sospechosos y probables (que no pudieron ser confirmados por MAT), demuestra que esta afección es endémica y se encuentra subregistrada en la provincia de Buenos Aires, y que la distancia y la falta de recursos serían factores determinantes de estas características.

Evaluación de tres PCR cuantitativas para la detección de leptospiras patógenas en animales domésticos en Nicaragua / Evaluation of three qPCR for the detection of pathogenic leptospires in domestic animals in Nicaragua

Resumen: Introducción. En Nicaragua es necesario estandarizar pruebas moleculares como la PCR en tiempo real (quantitative Polymerase Chain Reaction, qPCR) que mejoren el diagnóstico de leptospirosis en humanos y animales. Objetivo. Evaluar tres qPCR para la detección de leptospiras patógenas en animales domésticos de Nicaragua. Materiales y métodos. Se diseñaron cebadores para la amplificación del gen LipL32 en SYBR Green (SYBR Green-A) y TaqMan, y en otros descritos previamente (SYBR Green-B). Las secuencias de 12 cepas obtenidas de la base de datos del National Center for Biotechnology Information (NCBI) se alinearon para la búsqueda de sondas y cebadores. La sensibilidad analítica se determinó calculando el equivalente genómico detectable, se utilizaron 18 cepas de referencia para la sensibilidad diagnóstica y 28 controles negativos para la especificidad. Los métodos se aplicaron en 129 muestras de orina de animales domésticos. Resultados. En SYBR Green-A se obtuvo un límite de detección de cuatro equivalentes genómicos; en TaqMan, la sensibilidad fue del 94,4 % (IC95% 81,1-100,0). Con SYBR Green-A, se obtuvo una sensibilidad del 77,8 % (IC95% 55,8-99,8), en tanto que con SYBR Green-B fue del 61,1 % (IC95% 35,8-86,4). En las tres pruebas se logró una especificidad del 100 % (IC95% 98,2-100,0). El 26,4 % de las muestras de animales domésticos fueron positivas con SYBR Green-A y el 6,2 % con SYBR Green-B. Conclusiones. El SYBR Green-A presentó un límite de detección bajo, en tanto que las tres técnicas evaluadas mostraron alta especificidad, en tanto que la TaqMan tuvo la mayor sensibilidad.

Abstract: Introduction: Molecular biology diagnostic methods such as real-time PCR should be used in Nicaragua to improve the diagnosis of leptospirosis in humans and animals. Objective: To evaluate three qPCR methods for pathogenic Leptospira detection in domestic animals. Materials and methods: Real-time PCR primers were designed for the amplification of specific regions from the Lip 32 gene of Leptospira in SYBER Green (SYBER Green-A) and TaqMan, as well in SYBER Green-B as previously published. The sequences of 12 strains obtained from the database of the National Center for Biotechnology Information (NCBI) were aligned to select probes and primers. The analytical sensitivity was determined by calculating the detectable genomic equivalent while 18 pathogenic references strains and 28 negative controls were used to evaluate the sensitivity and specificity of each one of the three sets in 129 urine samples of domestic animals. Results: The detection limit of four genomic equivalents per reaction was obtained from SYBR Green-A. The specificities were 94.4% (95% CI: 81.1-100.0) for TaqMan, 77.8% (95% CI: 55.8-99.8) for SYBR Green-A, while for SYBR Green-B it was 61.1% (95% CI: 35.886.4). In the three tests, we obtained a specificity of 100% (95% CI: 98.2-100.0). In the field samples, 26.4% were positive with SYBR Green-A and 6.1% with SYBR Green-B. Conclusion: SYBR Green-A presented the lowest detection limit while the three techniques under evaluation showed high specificity while TaqMan was the most sensitive.

Primer aislamiento de Leptospira borgpetersenii serovar Hardjo tipo Hardjo Bovis a partir de un caso clínico en Argentina / First isolation of Leptospira borgpetersenii serovar Hardjo type Hardjo Bovis from aclinical case in cattle in Argentina

Resumen Se describe el primer aislamiento y la tipificación molecular de Leptospira borgpetersenii serovar Hardjo Bovis en Argentina, obtenido a partir de orina de vacas abortadas de unrodeo de cría ubicado en Saladillo, provincia de Buenos Aires. Los abortos coincidieron con unperíodo de importantes inundaciones, en el que varios animales presentaron títulos serológicossospechosos y posterior seroconversión. El porcentaje de abortos alcanzó el 3,5% del total delrodeo, compuesto por 1700 vacas, y se aisló el microorganismo en 7 de 20 muestras de orinaobtenidas.

Abstract We here describe the first isolation and molecular typing of Leptospira borgpe-tersenii serovar Hardjo Bovis in Argentina, obtained from urine of aborted cows from abreeding herd located in Saladillo, Buenos Aires Province. The abortions occurred in coincidence with important floods with many cows presenting suspicious serological titers and subsequentseroconversion. The percentage of abortions was 3.5% of a herd of 1700 cows and the microor-ganism was isolated from 7 of the 20 urine samples obtained.

Seroprevalence and incidence of Leptospira spp. in domestic dogs in the Southeast region of São Paulo State, Brazil / Soroprevalência e incidência de Leptospira spp. em cães domésticos na região sudeste do estado de São Paulo, Brasil

Leptospirosis is a zoonotic disease caused by Leptospira and domestic dogs can act as host of some serovars. In order to analyze the transmission dynamics in a dog population, with and without immunization, a longitudinal study was carried out with a focus to evaluate antibody response and to identify serovars. Blood samples were collected in three consecutive years (2015 to 2017) from 331, 373 and 347 dogs respectively. The dog seroprevalence in each year was 11%, 7% and 14%, respectively, and the incidence in 2016 was 5% and in 2017, 14%. The most frequent serovars were Cynopteri and Butembo in 2015, Cynopteri, Butembo and Hardjoprajitno in 2016, and Canicola and Butembo in 2017. Dogs can play a role as sentinel animals and hosts of Leptospira serovars. The percentage of seropositive dogs due to vaccination was higher than the previous years without immunization and lower than in previous years for other serovars, which we interpret as evidence for the importance of immunization. These parameters associated with active canine population control are important for prevention and control of leptospirosis not only in dogs but alsoto inhibit the transmission between dogs and humans.(AU)

A leptospirose é uma zoonose causada pelo agente etiológico Leptospira. Cães domésticos atuam como hospedeiro de diversos sorovares deste agente. Com intuito de analisar a dinâmica da leptospirose em uma população canina, com e sem imunização, um estudo longitudinal foi realizado avaliando a resposta sorológica destes animais e identificando seus sorovares. Foram coletadas amostras de 331, 373 e 347 cães em três anos consecutivos (2015 a 2017). As soroprevalências foram de 11%, 7% e 14%, respectivamente, e a incidência em 2016 foi de 5% e em 2017 de 14%. Os sorovares mais frequentes foram Cynopteri e Butembo em 2015, Cynopteri, Butembo e Hardjoprajitno em 2016, e Canicola e Butembo em 2017. Estes cães estão atuando como bio-indicadores da presença de Leptospira na região do estudo, incluindo sorovares zoonóticos, e contribuindo com a sua manutenção no ambiente. A soropositividade para sorovares protegidos pela vacina foi mais alta do que nos anos anteriores à imunização, enquanto para os sorovares não protegidos pela vacina diminuiu, demonstrando a importância da imunização para essa população de cães. Medidas de prevenção e controle para a leptospirose, como imunização e controle populacional canino, são recomendadas no local para inibir a transmissão do agente entre as populações de cães e humanos envolvidas.(AU)

Frequency of leptospirosis in horses in Manaus and metropolitan region in Amazonas State, Brazil / Frequência de leptospirose em equinos de Manaus e região metropolitana no estado do Amazonas, Brasil

The occurrence of leptospirosis is significantly higher in the tropical climate than in temperate regions, especially in periods with high rainfall levels. In horses, leptospirosis can range from subclinical to asymptomatic but maintains an important role in the transmission of leptospirosis by eliminating the agent in the environment. Regarding horses, the Amazonas herd increased by 30% between 2004 and 2013. Thus, the study aimed to determine the serological prevalence of leptospirosis in horses in Manaus and metropolitan regions, in the State of Amazonas, Brazil as well as to assess the main serogroups involved in the infections of these animals. For this purpose, from August 2018 to July 2019, serum from 198 horses was evaluated through the microscopic agglutination test with a panel of 10 serogroups. As a result, 92 horses (44.46%) were found to be positive for one or more Leptospira serovars, with the highest prevalence of serogroups Icterohaemorrhagiae and Pyrogenes. Therefore, with the increase in the equine herd in the region and, consequently, greater interaction between breeders and animals, the identification of reagents to at least one serovar of Leptospira spp. signals the possible existence of reservoirs of pathogenic strains for other animals and man.(AU)

A ocorrência de leptospirose é significativamente maior no clima tropical do que nas regiões temperadas, especialmente em períodos com altos níveis de chuva. Em cavalos, a leptospirose pode variar de subclínica a assintomática, mas estes animais podem desempenhar um papel importante na transmissão da leptospirose por meio da eliminação do agente no ambiente. Em relação aos cavalos, entre os anos de 2004 e 2013, o rebanho Amazônico aumentou 30%. Assim, o objetivo do estudo foi determinar a prevalência sorológica de leptospirose em cavalos em Manaus e região metropolitana, estado do Amazonas, Brasil, bem como, determinar os principais sorogrupos envolvidos nas infecções desses animais. Para este propósito, entre agosto de 2018 e julho de 2019, foram colhidos soros de 198 cavalos para serem avaliados sorologicamente com o teste de microaglutinação microscópica, utilizando-se um painel de dez sorogrupos. Como resultado, observou-se que 92 cavalos (44,46%) foram positivos para um ou mais sorovares de Leptospira, com maior prevalência dos sorogrupos Icterohaemorrhagiae e Pyrogenes. Portanto, com o aumento do rebanho equino na região e, consequentemente, maior interação entre criadores e animais, a identificação de reagentes para pelo menos um sorovar de Leptospira spp. alerta para a possível existência de reservatórios de cepas patogênicas para outros animais e seres humanos.(AU)

Canine leptospirosis in asymptomatic populations from the Southwest Region of São Paulo State, Brazil / Leptospirose canina em uma população assintomática da região sudoeste do estado de São Paulo, Brasil

Leptospirosis is an infectious disease that can affect animals and humans. Distributed worldwide, the disease is more prevalent in tropical regions due to socioenvironmental characteristics. Dogs can serve as sentinels for environmental contamination due to their frequent contact with humans. This study investigated the frequency of occurrence of canine leptospirosis in asymptomatic populations from the Southwest Region of the State of São Paulo, Brazil. Thus, blood samples collected from 572 asymptomatic dogs from the cities of Apiaí, Cananeia, Itapeva, and Itu were tested with a microscopic agglutination test (MAT). A total of 40.5% of animals in Apiaí reacted to Leptospira spp., 42.6% in Itapeva, 7.1% in Cananeia, and 5.1% in Itu. The data from the present study demonstrate that at least one animal from the municipalities of Itapeva, Apiaí, and Cananeia had a titer equal to or higher than 800, indicating that Leptospira is circulating in these municipalities and that the teams working on castration campaigns need to be educated on the correct use of personal protective equipment, especially when mechanically emptying the bladder of these animals. This study also suggests that castration campaigns can strategically monitor zoonotic diseases and assist in establishing preventive strategies for human and animal health.(AU)

A leptospirose é uma enfermidade infectocontagiosa que pode acometer os animais e o homem. Nos países tropicais e em desenvolvimento ocorrem 70% dos casos humanos, com mortalidade variando entre 10 a 70%. Os cães podem se tornar portadores assintomáticos por um longo período, podendo transmitir a Leptospira para humanos. Devido ao intenso convívio com o ser humano, os cães podem servir como sentinelas da contaminação ambiental. Esse trabalho investigou a frequência de ocorrência da leptospirose canina em populações assintomáticas da região sudoeste do estado de São Paulo. Para isso foram examinadas pela técnica de soroaglutinação microscópica (MAT), amostras de sangue provenientes de 572 cães assintomáticos dos municípios de Apiaí, Cananeia, Itapeva e Itu por amostragem de conveniência, oriundos de campanhas de castração. Em Apiaí, foram encontrados 40,5% dos animais reagentes para Leptospira spp.; em Itapeva, 42,6%; em Cananeia, 7,7% e em Itu, 5,1%. Os dados encontrados demonstram que, pelo menos, um animal dos municípios de Itapeva, Apiaí e Cananeia apresentaram título igual ou maior que 800, indicando a circulação da bactéria nessas localidades e que a equipe envolvida nas campanhas de castração precisam ser alertadas sobre o correto uso de equipamento de proteção individual, principalmente no esvaziamento mecânico da bexiga antes do procedimento cirúrgico. O estudo também sugere que as campanhas de castração podem ser estratégicas no monitoramento de doenças zoonóticas e poderiam auxiliar no estabelecimento de ações preventivas para a saúde humana e animal.(AU)

Fiebre amarilla selvática con serología positiva para leptospira en un varon joven peruano / Jungle yellow fever with positive serology for leptospira in a young peruvian male

RESUMEN Fiebre amarilla (FA) y leptospirosis son zoonosis endémicas subdiagnosticadas de las regiones tropicales de África y Sudamérica. Ambas, pueden ser clínicamente indistinguibles y presentarse como síndrome febril icterohemorrágico agudo. Reportamos el caso de un varón de 20 años, procedente del departamento de Amazonas que se presentó con nueve días de enfermedad caracterizada por falla multiorgánica (compromiso neurológico, renal, hepático, respiratorio, y hematológico). Recibió tratamiento antibiótico, soporte transfusional, dialítico, hemodinámico, y ventilatorio; y a pesar de la gravedad del cuadro clínico, evolucionó favorablemente. Se confirmó FA por Rt-PCR y se obtuvo serología positiva para leptospira por ELISA y microaglutinación. Sin embargo, no se pudo demostrar, desde el punto de vista laboratorial, coinfección real por FA y leptospira. Este caso de FA severa con desenlace no fatal enfatiza la importancia del diagnóstico sindrómico adecuado, y un tratamiento de soporte precoz y agresivo que puede salvar la vida del paciente.

ABSTRACT Yellow fever (YF) and leptospirosis are under-diagnosed endemic zoonoses of the tropical regions of Africa and South America. Both may be clinically indistinguishable and present as an acute icterohemorrhagic febrile syndrome. We report the case of a 20-year-old male from the department of Amazonas who presented with nine days of disease characterized by multiorgan failure (neurological, renal, hepatic, respiratory, and hematological involvement). He received antibiotic treatment, as well as, transfusion, dialysis, hemodynamic, and ventilatory support. Despite the severity of the clinical condition, he evolved favorably. YF was confirmed by Rt-PCR and positive serology was obtained for leptospira by ELISA and microagglutination. However, from a laboratory point of view, real co-infection by yellow fever and leptospira could not be demonstrated. This case of severe YF with non-fatal outcome emphasizes the importance of adequate syndromic diagnosis, and early and aggressive supportive treatment that can save a patient's life.