A One Health Approach to Investigating Leptospira Serogroups and Their Spatial Distributions among Humans and Animals in Rio Grande do Sul, Brazil, 2013⁻2015.

Leptospirosis is an endemic zoonotic disease in Brazil and is widespread throughout rural populations in the state of Rio Grande do Sul. This study aimed to identify presumptive infecting Leptospira serogroups in human and animal cases and describe their occurrences within the ecoregions of the state by animal species. Data for human and animal leptospirosis cases were gathered from the government's passive surveillance systems and presumptive infecting serogroups were identified based on a two-fold titer difference in serogroups in the microscopic agglutination test (MAT) panel. A total of 22 different serogroups were reported across both human and animal cases. Serogroup Icterohaemorrhagiae was the most common among humans, while serogroup Sejroe predominated among animal cases, particularly bovines. Each ecoregion had a large distribution of cases, with 51% of the human cases in the Parana⁻Paraiba ecoregion, and 81% of the animal cases in the Savannah ecoregion. Identifying and mapping the serogroups circulating using the One Health approach is the first step for further understanding the distribution of the disease in the state. This study has the potential to aid in guiding public health and agricultural practices, furthering the need for a human vaccine in high-risk populations to complement control and prevention efforts.

A road map for leptospirosis research and health policies based on country needs in Latin America.

This report summarizes the presentations, discussions and the recommendations coming from the Oswaldo Cruz Institute/FIOCRUZ International Workshop for Leptospirosis Research Based on Country Needs and the 5th Global Leptospirosis Environmental Action Network meeting, which was held in the city of Rio de Janeiro, Brazil, 10-12 November 2015. The event focused on health policy and worked to develop a road map as a consensus document to help guide decision-making by policymakers, funding bodies, and health care professionals. The direction that leptospirosis research should take in the coming years was emphasized, taking into account the needs of countries of Latin America, as well as experiences from other world regions, as provided by international experts. The operational concepts of "One Health" and translational research underlaid the discussions and the resulting recommendations. Despite the wide geographic distribution of leptospirosis and its impact in terms of incidence, morbidity, and mortality, leptospirosis is not yet considered a "tool-ready" disease for global initiatives. Surveillance programs need new tools and strategies for early detection, prevention, and follow-up. The major recommendations developed at the Rio meeting cover both health policy and research. The health policy recommendations should be taken into account by decisionmakers, government officials, and the Pan American Health Organization. The priorities for research, technological development, and innovation should be considered by research institutions, universities, and stakeholders.

A road map for leptospirosis research and health policies based on country needs in Latin America

This report summarizes the presentations, discussions and the recommendations coming from the Oswaldo Cruz Institute/FIOCRUZ International Workshop for Leptospirosis Research Based on Country Needs and the 5th Global Leptospirosis Environmental Action Network meeting, which was held in the city of Rio de Janeiro, Brazil, 10–12 November 2015. The event focused on health policy and worked to develop a road map as a consensus document to help guide decision-making by policymakers, funding bodies, and health care professionals. The direction that leptospirosis research should take in the coming years was emphasized, taking into account the needs of countries of Latin America, as well as experiences from other world regions, as provided by international experts. The operational concepts of “One Health” and translational research underlaid the discussions and the resulting recommendations. Despite the wide geographic distribution of leptospirosis and its impact in terms of incidence, morbidity, and mortality, leptospirosis is not yet considered a “tool-ready” disease for global initiatives. Surveillance programs need new tools and strategies for early detection, prevention, and follow-up. The major recommendations developed at the Rio meeting cover both health policy and research. The health policy recommendations should be taken into account by decisionmakers, government officials, and the Pan American Health Organization. The priorities for research, technological development, and innovation should be considered by research institutions, universities, and stakeholders.

En este informe se resumen las ponencias, los debates y las recomendaciones del taller internacional para la investigación sobre la leptospirosis basada en las necesidades de los países organizado por el Instituto Oswaldo Cruz/FIOCRUZ y la quinta reunión de la Red Global de Acción Ambiental contra la Leptospirosis, que se celebró en la ciudad del Rio de Janeiro (Brasil), del 10 al 12 de noviembre del 2015. El evento se centró en las políticas de salud y en la elaboración de una hoja de ruta que sirviese como un documento guía consensuado para la toma de decisiones de los formuladores de políticas, los organismos de financiamiento y los profesionales de la salud. Se hizo hincapié en la dirección que tiene que tomar la investigación sobre la leptospirosis en los próximos años, en la que se deben tener en cuenta las necesidades de los países de América Latina, así como a las experiencias de otras regiones del mundo, de acuerdo con las consideraciones de los expertos internacionales. Los conceptos operativos de “Una sola salud” y de la investigación traslacional fueron la base de los debates y de las recomendaciones resultantes. A pesar de la extensa distribución geográfica de la leptospirosis y su repercusión desde el punto de vista de la incidencia, la morbilidad y la mortalidad, la leptospirosis no se considera todavía una enfermedad con herramientas listas para ser aplicadas (tool-ready) en las iniciativas a nivel mundial. Los programas de vigilancia necesitan herramientas y estrategias nuevas para llevar a cabo la detección temprana, la prevención y el seguimiento. Las recomendaciones principales que se emitieron en la reunión de Rio cubrieron tanto el ámbito de las políticas de salud como el de la investigación. Las recomendaciones que se refieren a las políticas de salud deben ser tenidas en cuenta por los encargados de tomar decisiones, los funcionarios de gobierno y la Organización Panamericana de la Salud. Las prioridades de la investigación, el desarrollo tecnológico y la innovación deben ser incorporadas por las instituciones de investigación, las universidades y los interesados directos.

Trata-se de um resumo das apresentações, debates e recomendações do Workshop Internacional para Pesquisa em Leptospirose com Base nas Necessidades dos Países do Instituto Oswaldo Cruz/FIOCRUZ e 5a Reunião da Rede Global de Ação Ambiental em Leptospirose, realizados na cidade do Rio de Janeiro, Brasil, de 10 a 12 de novembro de 2015. Os participantes do evento debateram políticas de saúde e elaboraram um guia como documento de consenso para orientar a tomada de decisão pelos responsáveis por políticas, agências de financiamento e profissionais da saúde. Destacou-se o rumo a ser tomado em pesquisa em leptospirose nos próximos anos levando em consideração as necessidades dos países da América Latina, assim como as experiências de outras regiões do mundo, conforme estabelecido por especialistas internacionais. Os conceitos práticos da perspectiva “One Health” (Um Mundo, Uma Saúde) e pesquisa translacional foram a base dos debates e das recomendações conseguintes. Apesar da ampla distribuição geográfica e do seu impacto em termos de incidência, morbidade e mortalidade, a leptospirose ainda não é considerada uma doença “tool ready” para iniciativas globais. Os programas de vigilância precisam de novas ferramentas e estratégias para detecção precoce, prevenção e acompanhamento. As principais recomendações elaboradas na reunião do Rio englobam política de saúde assim como pesquisa e elas devem ser levadas em consideração pelas autoridades, funcionários do governo e Organização Pan-Americana da Saúde. As instituições de pesquisa, universidades e interessados diretos devem atentar para as prioridades em pesquisa, desenvolvimento tecnológico e inovação.

Molecular and serological characterization of Leptospira kirschneri serogroup Pomona isolated from a human case in a Brazilian rural area.

INTRODUCTION:: Leptospirosis is an important health concern in Brazil. Currently, information on the epidemiology of the disease in the rural areas of the country is lacking. METHODS:: Serological and molecular techniques were used to characterize a clinical isolate of Leptospira. RESULTS:: The strain CLEP 00060, isolated from a 59-year-old man in a rural area of Rio Grande do Sul state, Brazil, was identified as belonging to L. kirschneri serogroup Pomona serovar Mozdok. CONCLUSIONS:: This study contributes to the local epidemiological knowledge of leptospirosis, prevention of the disease by vaccines, and improvements in its diagnosis.

Molecular and serological characterization of Leptospira kirschneri serogroup Pomona isolated from a human case in a Brazilian rural area

Abstract INTRODUCTION: Leptospirosis is an important health concern in Brazil. Currently, information on the epidemiology of the disease in the rural areas of the country is lacking. METHODS: Serological and molecular techniques were used to characterize a clinical isolate of Leptospira. RESULTS: The strain CLEP 00060, isolated from a 59-year-old man in a rural area of Rio Grande do Sul state, Brazil, was identified as belonging to L. kirschneri serogroup Pomona serovar Mozdok. CONCLUSIONS: This study contributes to the local epidemiological knowledge of leptospirosis, prevention of the disease by vaccines, and improvements in its diagnosis.

High-Resolution Melting Curve Analysis of the 16S Ribosomal Gene to Detect and Identify Pathogenic and Saprophytic Leptospira Species in Colombian Isolates.

AbstractIt is important to identify the circulating Leptospira agent to enhance the performance of serodiagnostic tests by incorporating specific antigens of native species, develop vaccines that take into account the species/serovars circulating in different regions, and optimize prevention and control strategies. The objectives of this study were to develop a polymerase chain reaction (PCR)-high-resolution melting (HRM) assay for differentiating between species of the genus Leptospira and to verify its usefulness in identifying unknown samples to species level. A set of primers from the initial region of the 16S ribosomal gene was designed to detect and differentiate the 22 species of Leptospira. Eleven reference strains were used as controls to establish the reference species and differential melting curves. Twenty-five Colombian Leptospira isolates were studied to evaluate the usefulness of the PCR-HRM assay in identifying unknown samples to species level. This identification was confirmed by sequencing and phylogenetic analysis of the 16S ribosomal gene. Eleven Leptospira species were successfully identified, except for Leptospira meyeri/Leptospira yanagawae because the sequences were 100% identical. The 25 isolates from humans, animals, and environmental water sources were identified as Leptospira santarosai (twelve), Leptospira interrogans (nine), and L. meyeri/L. yanagawae (four). The species verification was 100% concordant between PCR-HRM and phylogenetic analysis of the 16S ribosomal gene. The PCR-HRM assay designed in this study is a useful tool for identifying Leptospira species from isolates.

Leptospirosis in Latin America: exploring the first set of regional data

Objectives. To demonstrate the importance of country surveillance systems for leptospirosis and their use for preliminary epidemiological analysis, as well as to generate research questions for future, more comprehensive studies on the disease. Methods. In 2015, for the first time, the Pan American Health Organization (PAHO) included human cases of leptospirosis in its Regional Core Health Data Initiative, an open-access database that collects annual health indicators from the countries and territories of the Americas. This new information was used to analyze leptospirosis cases by country and sex and to calculate cumulative incidence rates. Maps were used to help present the results. To supplement that general review of leptospirosis in the Americas, more detailed descriptions of the epidemiological situation and the surveillance programs of four selected countries (Brazil, Colombia, Cuba, and Honduras) were provided. Results. In this first year of PAHO requesting leptospirosis data, of the 49 countries and territories in the Americas, 38 of them (77.6%) reported information. Among those 38, 28 of them (73.7%) reported the presence of human cases; the majority of instances of zero cases were in Caribbean territories. From those 28, a total of 10 702 human cases were recorded. The largest numbers of cases in Latin America were in Brazil (40.2%), Peru (23.6%), Colombia (8.8%), and Ecuador (7.2%). The cumulative incidence rate for Latin America was estimated to be 2.0 per 100 000 population. On average, 65.1% of cases were males. Conclusions. This study demonstrates that many countries in Latin America are making efforts to establish strong surveillance systems and programs for leptospirosis. The study also shows the importance of having leptospirosis surveillance systems as well as how the information generated can be used for evidence-based decision-making on leptospirosis.

Objetivos. Demostrar la importancia de los sistemas nacionales de vigilancia de la leptospirosis y su uso para realizar el análisis epidemiológico preliminar, así como para generar preguntas de investigación que se utilicen en futuros estudios más integrales sobre la enfermedad. Métodos. En el año 2015, la Organización Panamericana de la Salud (OPS) incluyó por primera vez casos humanos de leptospirosis en su Iniciativa Regional de Datos Básicos de Salud, base de datos de acceso abierto que recoge indicadores de salud anuales de los países y territorios de la Región de las Américas. Esta nueva información fue utilizada para analizar los casos de leptospirosis por país y sexo, así como para calcular las tasas de incidencia acumulada, y los resultados se presentaron en mapas. Para complementar ese examen general de la leptospirosis en esta Región, se aportó una descripción más detallada de la situación epidemiológica y los programas de vigilancia de cuatro países (Brasil, Colombia, Cuba y Honduras). Resultados. En este primer año en que la OPS solicitó datos sobre la leptospirosis, de los 49 países y territorios en la Región de las Américas, aportaron información 38 (77,6 %). De esos 38, 28 (73,7 %) notificaron casos humanos; la mayor parte de las instancias sin ningún caso humano se registraron en territorios del Caribe. En esos 28 países y territorios, se registraron 10 702 casos humanos. En América Latina, los países que registraron los números más altos de casos fueron Brasil (40,2 %), Perú (23,6 %), Colombia (8,8 %) y Ecuador (7,2 %). Se calculó que la tasa de incidencia acumulada de América Latina es de 2,0 por 100 000 habitantes. En promedio, 65,1 % de los casos correspondieron a hombres. Conclusiones. Este estudio demuestra que muchos países de América Latina están tomando medidas para instaurar sistemas y programas sólidos de vigilancia de la leptospirosis. Asimismo, revela la importancia de los sistemas de vigilancia de la leptospirosis, así como el modo en que la información generada puede servir para tomar decisiones basadas en la evidencia acerca de esta enfermedad.

Objetivos. Demonstrar a importância dos sistemas nacionais de vigilância para a leptospirose e seu uso na análise epidemiológica preliminar, além de gerar perguntas de pesquisa para futuros estudos mais aprofundados sobre a doença. Métodos. Em 2015, a Organização Pan-Americana da Saúde (OPAS) incluiu, pela primeira vez, casos humanos de leptospirose em sua Iniciativa Regional de Dados Básicos de Saúde, uma base de dados de acesso aberto que coleta indicadores de saúde anuais nos países e territórios das Américas. Utilizamos estas novas informações para analisar os casos de leptospirose por país e por sexo e calcular as taxas de incidência acumuladas. A apresentação dos dados é facilitada pelo uso de mapas. Para complementar esta revisão geral da leptospirose nas Américas, apresentamos descrições mais detalhadas da situação epidemiológica e dos programas de vigilância de quatro países selecionados (Brasil, Colômbia, Cuba e Honduras). Resultados. Neste primeiro ano em que a OPAS solicitou dados sobre a leptospirose, 38 dos 49 países e territórios das Américas (77,6%) apresentaram informações. Destes 38, 28 (73,7%) notificaram a presença de casos humanos; em sua maioria, os que não apresentaram nenhum caso foram territórios do Caribe. Dentre os 28, foi registrado um total de 10.702 casos humanos. O maior número de casos na América Latina foi observado no Brasil (40,2%), Peru (23,6%), Colômbia (8,8%) e Equador (7,2%). A taxa de incidência acumulada para a América Latina foi estimada em 2,0 por 100.000 habitantes. Em média, 65,1% dos casos foram em homens. Conclusões. Este estudo demonstra que muitos países da América Latina estão fazendo esforços para estabelecer fortes sistemas de vigilância e programas contra a leptospirose. O estudo também revela a importância da existência de sistemas de vigilância para a leptospirose e demonstra que as informações geradas podem ser usadas para a tomada de decisões baseadas em evidências científicas para a leptospirose.

Leptospirosis in Latin America: exploring the first set of regional data / La leptospirosis en América Latina: exploración del primer conjunto de datos regionales / Leptospirose na América Latina: avaliação do primeiro conjunto de dados regionais

ABSTRACT Objectives To demonstrate the importance of country surveillance systems for leptospirosis and their use for preliminary epidemiological analysis, as well as to generate research questions for future, more comprehensive studies on the disease. Methods In 2015, for the first time, the Pan American Health Organization (PAHO) included human cases of leptospirosis in its Regional Core Health Data Initiative, an open-access database that collects annual health indicators from the countries and territories of the Americas. This new information was used to analyze leptospirosis cases by country and sex and to calculate cumulative incidence rates. Maps were used to help present the results. To supplement that general review of leptospirosis in the Americas, more detailed descriptions of the epidemiological situation and the surveillance programs of four selected countries (Brazil, Colombia, Cuba, and Honduras) were provided. Results In this first year of PAHO requesting leptospirosis data, of the 49 countries and territories in the Americas, 38 of them (77.6%) reported information. Among those 38, 28 of them (73.7%) reported the presence of human cases; the majority of instances of zero cases were in Caribbean territories. From those 28, a total of 10 702 human cases were recorded. The largest numbers of cases in Latin America were in Brazil (40.2%), Peru (23.6%), Colombia (8.8%), and Ecuador (7.2%). The cumulative incidence rate for Latin America was estimated to be 2.0 per 100 000 population. On average, 65.1% of cases were males. Conclusions This study demonstrates that many countries in Latin America are making efforts to establish strong surveillance systems and programs for leptospirosis. The study also shows the importance of having leptospirosis surveillance systems as well as how the information generated can be used for evidence-based decision-making on leptospirosis.

RESUMEN Objetivos Demostrar la importancia de los sistemas nacionales de vigilancia de la leptospirosis y su uso para realizar el análisis epidemiológico preliminar, así como para generar preguntas de investigación que se utilicen en futuros estudios más integrales sobre la enfermedad. Métodos En el año 2015, la Organización Panamericana de la Salud (OPS) incluyó por primera vez casos humanos de leptospirosis en su Iniciativa Regional de Datos Básicos de Salud, base de datos de acceso abierto que recoge indicadores de salud anuales de los países y territorios de la Región de las Américas. Esta nueva información fue utilizada para analizar los casos de leptospirosis por país y sexo, así como para calcular las tasas de incidencia acumulada, y los resultados se presentaron en mapas. Para complementar ese examen general de la leptospirosis en esta Región, se aportó una descripción más detallada de la situación epidemiológica y los programas de vigilancia de cuatro países (Brasil, Colombia, Cuba y Honduras). Resultados En este primer año en que la OPS solicitó datos sobre la leptospirosis, de los 49 países y territorios en la Región de las Américas, aportaron información 38 (77,6 %). De esos 38, 28 (73,7 %) notificaron casos humanos; la mayor parte de las instancias sin ningún caso humano se registraron en territorios del Caribe. En esos 28 países y territorios, se registraron 10 702 casos humanos. En América Latina, los países que registraron los números más altos de casos fueron Brasil (40,2 %), Perú (23,6 %), Colombia (8,8 %) y Ecuador (7,2 %). Se calculó que la tasa de incidencia acumulada de América Latina es de 2,0 por 100 000 habitantes. En promedio, 65,1 % de los casos correspondieron a hombres. Conclusiones Este estudio demuestra que muchos países de América Latina están tomando medidas para instaurar sistemas y programas sólidos de vigilancia de la leptospirosis. Asimismo, revela la importancia de los sistemas de vigilancia de la leptospirosis, así como el modo en que la información generada puede servir para tomar decisiones basadas en la evidencia acerca de esta enfermedad.

RESUMO Objetivos Demonstrar a importância dos sistemas nacionais de vigilância para a leptospirose e seu uso na análise epidemiológica preliminar, além de gerar perguntas de pesquisa para futuros estudos mais aprofundados sobre a doença. Métodos Em 2015, a Organização Pan-Americana da Saúde (OPAS) incluiu, pela primeira vez, casos humanos de leptospirose em sua Iniciativa Regional de Dados Básicos de Saúde, uma base de dados de acesso aberto que coleta indicadores de saúde anuais nos países e territórios das Américas. Utilizamos estas novas informações para analisar os casos de leptospirose por país e por sexo e calcular as taxas de incidência acumuladas. A apresentação dos dados é facilitada pelo uso de mapas. Para complementar esta revisão geral da leptospirose nas Américas, apresentamos descrições mais detalhadas da situação epidemiológica e dos programas de vigilância de quatro países selecionados (Brasil, Colômbia, Cuba e Honduras). Resultados Neste primeiro ano em que a OPAS solicitou dados sobre a leptospirose, 38 dos 49 países e territórios das Américas (77,6%) apresentaram informações. Destes 38, 28 (73,7%) notificaram a presença de casos humanos; em sua maioria, os que não apresentaram nenhum caso foram territórios do Caribe. Dentre os 28, foi registrado um total de 10.702 casos humanos. O maior número de casos na América Latina foi observado no Brasil (40,2%), Peru (23,6%), Colômbia (8,8%) e Equador (7,2%). A taxa de incidência acumulada para a América Latina foi estimada em 2,0 por 100.000 habitantes. Em média, 65,1% dos casos foram em homens. Conclusões Este estudo demonstra que muitos países da América Latina estão fazendo esforços para estabelecer fortes sistemas de vigilância e programas contra a leptospirose. O estudo também revela a importância da existência de sistemas de vigilância para a leptospirose e demonstra que as informações geradas podem ser usadas para a tomada de decisões baseadas em evidências científicas para a leptospirose.

Genetic diversity of Leptospira in northwestern Colombia: first report of Leptospira santarosai as a recognised leptospirosis agent.

The region of Antioquia in northeastern Colombia has the highest number of reported leptospirosis cases in the country. It also shows high seroprevalence indexes in the general population and socio-environmental conditions favourable for the transmission of the disease between humans and animals. In this study, 25 Leptospira isolates from Colombia's Antioquia department were identified to the species level as L. santarosai (12), L. interrogans (9) and L. meyeri (4) using phylogenetic analysis of the Amidohydrolase gene. Typing at the serovar level was performed using multilocus sequence typing (MLST) and monoclonal antibodies. The serovars Canalzonae, Babudieri, Alice, Beye, and Copenhageni have been identified as causing human or animal infections in Antioquia, Colombia. The four environmental isolates were not identified to the serovar level. L. santarosai serovar Canalzonae and Alice were identified as new etiologic agents of human leptospirosis in Antioquia, Colombia. This paper reports species and serovars that were previously unknown in the region.

Genetic diversity of Leptospira in northwestern Colombia: first report of Leptospira santarosai as a recognised leptospirosis agent

The region of Antioquia in northeastern Colombia has the highest number of reported leptospirosis cases in the country. It also shows high seroprevalence indexes in the general population and socio-environmental conditions favourable for the transmission of the disease between humans and animals. In this study, 25 Leptospira isolates from Colombia’s Antioquia department were identified to the species level as L. santarosai (12), L. interrogans (9) and L. meyeri (4) using phylogenetic analysis of the Amidohydrolase gene. Typing at the serovar level was performed using multilocus sequence typing (MLST) and monoclonal antibodies. The serovars Canalzonae, Babudieri, Alice, Beye, and Copenhageni have been identified as causing human or animal infections in Antioquia, Colombia. The four environmental isolates were not identified to the serovar level. L. santarosai serovar Canalzonae and Alice were identified as new etiologic agents of human leptospirosis in Antioquia, Colombia. This paper reports species and serovars that were previously unknown in the region.

Animal leptospirosis in Latin America and the Caribbean countries: reported outbreaks and literature review (2002-2014).

Leptospirosis is a worldwide zoonotic disease whose transmission is linked through multiple factors in the animal-human-ecosystem interface. The data on leptospirosis reported to the World Organization for Animal Health (OIE) for Latin America and Caribbean (LAC) countries/sovereign territories from 2005-2011 were mapped, showing a wide distribution of outbreaks in the region. Tropical terrestrial biomes are the predominate ecosystems showing reports of outbreaks. Climatic and ecological factors were relevant to the occurrence of epidemic outbreaks. The available scientific information from 2002-2014 was summarized to obtain a general overview and identify key issues related to the One Health approach. The primary serological test used for diagnosis and for conducting surveys was the microscopic agglutination test (MAT). Reports regarding the isolation and typing of leptospires were scattered and limited to data from a few countries, but their results revealed considerable biodiversity at the species and serovar levels. A total of six out of 11 currently named pathogenic species were found in the region. There was also high diversity of animal species showing evidence of infection by leptospires, including rodents, pets, livestock and wild animals. Prevention and control measures for leptospirosis should consider issues of animal and human health in the context of ecosystems, the territorial land borders of countries and trade.

Leptospirosis diagnosis by immunocapture polymerase chain reaction: a new tool for early diagnosis and epidemiologic surveillance.

The aim of this study was to develop an immunocapture polymerase chain reaction (IC-PCR) protocol for leptospirosis. For the standardization of IC-PCR, polyclonal (AS) and monoclonal (MAb) antibodies against different serogroups and serovars of Leptospira were coupled to polystyrene plates. Human sera were artificially contaminated with leptospires and incubated on plates. The bacterial DNA was obtained and used in a multiplex PCR. Sensitivity was tested using sera contaminated with crescent concentrations of leptospires, while specificity was established using sera contaminated with different bacterial genera and sera obtained from patients positive for viral infections. IC-PCR using AS was able to recognize specific serogroups, although some cross-reactions have been observed. No cross-reactions were observed when MAbs were used; however, the sensitivity in this case was lower than that of IC-PCR using AS. IC-PCR proved to be specific to Leptospira and is a promising tool for early diagnosis of leptospirosis, providing additional information about the infecting serovar or serogroup.

Leptospira and Brucella antibodies in collared anteaters (Tamandua tetradactyla) in Brazilian zoos.

The presence of Leptospira spp. and Brucella spp. antibodies was investigated in serum samples from 28 collared anteaters (Tamandua tetradactyla) kept in seven Brazilian zoos. Sera were tested against 19 Leptospira serovars using microscopic agglutination. Samples reacted to the following serovars: two (7.14%) to Patoc, three (10.71%) to Tarrasovi, three (10.71%) to both Patoc and Tarrasovi, two (7.14%) to Wolffi, and one (3.57%) to Australis. Two (7.14%) samples reacted to the buffered Brucella antigen test, but no confirmatory reaction occurred using the 2-mercaptoethanol slow slide agglutination test. No sample was reactive in the agar gel immunodiffusion test for rugose species of Brucella. The presence of anti-leptospira agglutinins in captive T. tetradactyla serum indicates that this species may be susceptible to infection by these bacteria.

Atypical lymphocytosis in leptospirosis: a cohort of hospitalized cases between 1996 and 2009 in State of Rio de Janeiro, Brazil / Linfócitos atípicos na leptospirose: coorte de pacientes hospitalizados entre 1996 e 2009, Estado do Rio de Janeiro

INTRODUCTION: Leptospirosis is a zoonotic disease found in tropical and temperate countries, and its clinical diagnostic confusion with arboviruses (dengue fever, oropouche fever and yellow fever), Brazilian spotted fever, viral hepatitis and hantaviruses has been an ongoing public health concern. The aim of this observational study was to demonstrate an association between findings of atypical lymphocytosis and the progression of endemic leptospirosis. METHODS: A retrospective analysis was performed on the demographic, epidemiological, clinical and laboratory aspects of 27 human leptospirosis cases that occurred over a period of 13 years (1996-2009) with no reported epidemic outbreaks in Rio de Janeiro, Brazil. RESULTS: The overall mortality rate was 11.1 percent in our cohort of hospitalized cases. However, there was no mortality among patients with atypical lymphocytosis (OR = 11.1; 95 percent CI = 1.12-110.9; p = 0.04). Two patients who were in the septicemic phase showed signs of expansion of γδ T cell responses in peripheral blood. CONCLUSIONS: Atypical lymphocytosis may be observed in patients with leptospirosis. Our observations suggest that these atypical leukocyte subsets are associated with partial protection during the disease course of leptospirosis.

INTRODUÇÃO: Leptospirose é uma zoonose que permanece endêmica em regiões tropicais e temperadas. A dificuldade no diagnóstico clínico diferencial entre os quadros de leptospirose humana e as várias arboviroses (dengue, febre amarela, febre de oropouche), febre maculosa brasileira, hepatite viral e hantavirose permanece um problema na Saúde Pública. MÉTODOS: No presente estudo, foi realizada análise retrospectiva de características demográficas, epidemiológicas, clínicas e laboratoriais de 27 casos de leptospirose humana que ocorrerem durante um período de 13 anos sem ocorrência de notificação de surtos epidêmicos no Rio de Janeiro, Brasil (1996-2009). RESULTADOS: A mortalidade da coorte de pacientes com leptospirose correspondeu a 11,1 por cento, sem embargo, o grupo de pacientes com atipia linfocitária não evoluiu para o óbito (OR = 11,1; 95 por cento CI = 1,12-110,9; p = 0.04). Em duas oportunidades, foi observada uma expansão dos linfócitos T gama-delta no sangue periférico de pacientes na fase septicêmica da leptospirose. CONCLUSÕES: Atipia linfocitária pode ocorrer em pacientes com leptospirose. Nossos dados também sugerem que os linfócitos atípicos podem estar envolvidos na patogênese da leptospirose.

Atypical lymphocytosis in leptospirosis: a cohort of hospitalized cases between 1996 and 2009 in State of Rio de Janeiro, Brazil.

INTRODUCTION: Leptospirosis is a zoonotic disease found in tropical and temperate countries, and its clinical diagnostic confusion with arboviruses (dengue fever, oropouche fever and yellow fever), Brazilian spotted fever, viral hepatitis and hantaviruses has been an ongoing public health concern. The aim of this observational study was to demonstrate an association between findings of atypical lymphocytosis and the progression of endemic leptospirosis. METHODS: A retrospective analysis was performed on the demographic, epidemiological, clinical and laboratory aspects of 27 human leptospirosis cases that occurred over a period of 13 years (1996-2009) with no reported epidemic outbreaks in Rio de Janeiro, Brazil. RESULTS: The overall mortality rate was 11.1% in our cohort of hospitalized cases. However, there was no mortality among patients with atypical lymphocytosis (OR = 11.1; 95% CI = 1.12-110.9; p = 0.04). Two patients who were in the septicemic phase showed signs of expansion of γδ T cell responses in peripheral blood. CONCLUSIONS: Atypical lymphocytosis may be observed in patients with leptospirosis. Our observations suggest that these atypical leukocyte subsets are associated with partial protection during the disease course of leptospirosis.

Multiplex PCR-based detection of Leptospira in environmental water samples obtained from a slum settlement.

The aim of this study was to apply a molecular protocol to detect leptospiral DNA in environmental water samples. The study was carried out in a peri-urban settlement in Petrópolis, state of Rio de Janeiro. A multiplex PCR method employing the primers LipL32 and 16SrRNA was used. Three out of 100 analysed samples were positive in the multiplex PCR, two were considered to have saprophytic leptospires and one had pathogenic leptospires. The results obtained supported the idea that multiplex PCR can be used to detect Leptospira spp in water samples. This method was also able to differentiate between saprophytic and pathogenic leptospires and was able to do so much more easily than conventional methodologies.

[Characterization of Leptospira sp reference strains using the pulsed field gel electrophoresis technique]. / Caracterização de cepas de referência de Leptospira sp utilizando a técnica de pulsed field gel electrophoresis.

INTRODUCTION: Leptospirosis is an endemic zoonosis of worldwide distribution, caused by bacteria of the genus Leptospira. This genus includes pathogenic and saprophytic species, with more than 200 different serovars, thus making it difficult to characterize. The technique of pulsed field gel electrophoresis has been used as a tool to aid in this characterization. The aims of this study were to standardize the PFGE technique, determine the molecular profiles of reference strains used at the National Reference Laboratory for Leptospirosis/World Health Organization Collaborating Center for Leptospirosis and create a database with these profiles. METHODS: Nineteen strains were analyzed by means of PFGE, using the restriction enzyme NotI. RESULTS: Each strain presented a unique profile that could be considered to be a specific genomic identity, with the exception of the serovars Icterohaemorrhagiae and Copenhageni, whose profiles were indistinguishable. CONCLUSIONS: It was possible to create a database of molecular profiles, which are being used in the Laboratory for comparing and identifying strains isolated from clinical cases.

Multiplex PCR-based detection of Leptospira in environmental water samples obtained from a slum settlement

The aim of this study was to apply a molecular protocol to detect leptospiral DNA in environmental water samples. The study was carried out in a peri-urban settlement in Petrópolis, state of Rio de Janeiro. A multiplex PCR method employing the primers LipL32 and 16SrRNA was used. Three out of 100 analysed samples were positive in the multiplex PCR, two were considered to have saprophytic leptospires and one had pathogenic leptospires. The results obtained supported the idea that multiplex PCR can be used to detect Leptospira spp in water samples. This method was also able to differentiate between saprophytic and pathogenic leptospires and was able to do so much more easily than conventional methodologies.

Caracterização de cepas de referência de Leptospira sp utilizando a técnica de pulsed field gel electrophoresis / Characterization of Leptospira sp reference strains using the pulsed field gel electrophoresis technique

INTRODUÇÃO: A leptospirose é uma zoonose endêmica, mundialmente distribuída, causada por bactérias do gênero Leptospira. Este gênero compreende espécies patogênicas e saprofíticas, com mais de 200 sorovares distintos, dificultando sua caracterização. A técnica de pulsed field gel electrophoresis tem sido empregada como uma ferramenta para auxiliar nesta caracterização. Os objetivos deste trabalho foram padronizar a técnica de PFGE, determinar os perfis moleculares das cepas de referência utilizadas pelo Laboratório de Referência Nacional para Leptospirose/Centro Colaborador da Organização Mundial de Saúde para Leptospirose e criar um banco de dados com estes perfis. MÉTODOS: Foram analisadas, por PFGE, dezenove cepas utilizando a enzima de restrição NotI. RESULTADOS: Cada cepa apresentou um perfil único que pode ser considerado como uma identidade genômica específica, com exceção dos sorovares Icterohaemorrhagiae e Copenhageni, cujos perfis foram indistinguíveis. CONCLUSÕES: Dessa forma, foi possível a criação de um banco de perfis moleculares que está sendo utilizado no Laboratório para a comparação e identificação de cepas isoladas de quadros clínicos.

INTRODUCTION: Leptospirosis is an endemic zoonosis of worldwide distribution, caused by bacteria of the genus Leptospira. This genus includes pathogenic and saprophytic species, with more than 200 different serovars, thus making it difficult to characterize. The technique of pulsed field gel electrophoresis has been used as a tool to aid in this characterization. The aims of this study were to standardize the PFGE technique, determine the molecular profiles of reference strains used at the National Reference Laboratory for Leptospirosis/World Health Organization Collaborating Center for Leptospirosis and create a database with these profiles. METHODS: Nineteen strains were analyzed by means of PFGE, using the restriction enzyme NotI. RESULTS: Each strain presented a unique profile that could be considered to be a specific genomic identity, with the exception of the serovars Icterohaemorrhagiae and Copenhageni, whose profiles were indistinguishable. CONCLUSIONS: It was possible to create a database of molecular profiles, which are being used in the Laboratory for comparing and identifying strains isolated from clinical cases.

Carriage of Leptospira interrogans among domestic rats from an urban setting highly endemic for leptospirosis in Brazil.

A survey was conducted to identify reservoirs for urban leptospirosis in the city of Salvador, Brazil. Sampling protocols were performed in the vicinity of households of severe leptospirosis cases identified during active hospital-based surveillance. Among a total of 142 captured Rattus norvegicus (Norwegian brown rat), 80.3% had a positive culture isolate from urine or kidney specimens and 68.1% had a positive serum sample by microscopic agglutination test (MAT) titre of > or = 1:100. Monoclonal antibody-based typing of isolates identified that the agent carried by rats was Leptospira interrogans serovar Copenhageni, which was the same serovar isolated from patients during hospital-based surveillance. Leptospira spp. were not isolated from 8 captured Didelphis marsupialis (Opossum), while 5/7 had a positive MAT titre against a saprophytic serogroup. R. rattus were not captured during the survey. The study findings indicate that the brown rat is a major rodent reservoir for leptospirosis in this urban setting. Furthermore, the high carriage rates of L. interrogans serovar Copenhageni in captured rats suggest that there is a significant degree of environmental contamination with this agent in the household environment of high risk areas, which in turn is a cause of transmission during urban epidemics.