Leptospirosis: predictores de mala evolución clínica en pacientes hospitalizados, 25 años de experiencia / Leptospirosis: predictors of poor outcomes in hospitalized patients, 25 years of experience

Introducción: La leptospirosis es una zoonosis que cons-tituye un problema emergente de salud pública. La insufi-ciencia renal, plaquetopenia y compromiso respiratorio se describen como predictores de mortalidad.Objetivos: Describir características clínicas, radiológicas y de laboratorio de individuos hospitalizados por leptos-pirosis y evaluar los predictores de mala evolución clínica (MEC).Materiales y métodos: Estudio de cohorte de inclusión ambispectiva de pacientes con leptospirosis internados en un hospital de la ciudad de Santa Fe entre 1997 y 2022. Se definió MEC como la admisión a Unidad de Cuidados Intensivos (UCI), requerimiento de asistencia respiratoria mecánica (ARM) y/o muerte. Se utilizaron las pruebas de Chi2, test T de Student o la U de Mann-Whitney, según co-rrespondiera. Se construyó una regresión logística binaria con las variables con p<0,05.Resultados: 101 pacientes, 87,1% (n=88) hombres, media-na de edad de 29 (RIC 20-44) años. La fiebre fue el síntoma más frecuente [83,2% (n=84)], seguido del compromiso di-gestivo [62,4% (n=63)]. Las alteraciones de laboratorio más frecuentes fueron: eritrosedimentación elevada [91,9% (n=79)] y leucocitosis [61% (n=61)]. Se observó MEC en el 25,7% (n=26). El 25,7% (n=26) fue admitido en UCI, el 13,9% (n=14) requirió ARM y el 5% (n=5) falleció. La presencia de plaquetopenia (OR=13,3, IC95% 2-80), las alteraciones en la radiografía de tórax (OR=33,5, IC95% 5-225) y la ausencia de cefalea (OR=6,8, IC95% 1-32) fueron predictores inde-pendientes de MEC.Conclusiones: En concordancia con la bibliografía, la afec-tación pulmonar y plaquetopenia son factores de riesgo para la mala evolución clínica. En nuestra serie, la cefalea constituyó un síntoma protector

Introduction: Leptospirosis is an emerging zoonotic di-sease that poses a public health problem. Renal failu-re, thrombocytopenia, and respiratory involvement have been described as predictors of mortality.Objectives: To describe the clinical, radiological, and la-boratory characteristics of hospitalized individuals with leptospirosis and evaluate predictors of poor clinical outcomes (PCO).Materials and methods: A prospective cohort study was conducted including patients with leptospirosis admit-ted to a hospital in the city of Santa Fe between 1997 and 2022. PCO was defined as admission to the Intensive Care Unit (ICU), requirement for mechanical respiratory assistance (MRA), and/or death. The chi-square test, Student>s t-test, or Mann-Whitney U test were used as appropriate. A binary logistic regression was performed with variables having p<0.05.Results: Out of the 101 patients included, 87.1% (n=88) were male, with a median age of 29 (IQR 20-44) years. Fever was the most common symptom [83.2% (n=84)], followed by digestive involvement [62.4% (n=63)]. The most frequent laboratory abnormalities were elevated erythrocyte sedimentation rate [91.9% (n=79)] and leuko-cytosis [61% (n=61)]. PCO was observed in 25.7% (n=26) of patients, with 25.7% (n=26) admitted to the ICU, 13.9% (n=14) requiring MRA, and 5% (n=5) resulting in death. The presence of thrombocytopenia (OR=13.3, 95% CI 2-80), abnormalities in chest X-rays (OR=33.5, 95% CI 5-225), and absence of headache (OR=6.8, 95% CI 1-32) were predictors of PCO. Conclusions: Consistent with the literature, pulmonary involvement and thrombocytopenia are independent risk factors for poor clinical outcomes. In our series, the pre-sence of headache was a protective symptom

LEPTOSPIROSIS: A NEGLECTED DISEASE FROM NORTH INDIA

Introduction: Leptospirosis is a zoonotic disease caused by Leptospira interrogans and has been reported from various countries worldwide. As very few studies were conducted on leptospirosis from north India, this study was conducted to know the status of this disease in this region. This retrospective hospital Material & Methods: based study was conducted in the Department of Microbiology of a tertiary care super specialty teaching institute from north India for a period of two consecutive years. Blood specimens from acute febrile illness cases were tested for presence of IgM antibodies against Leptospira interrogans by rapid card (Leptocheck from TULIP) testing and ELISA (Leptospira IgM ELISA from PanBio). Out of total 216 samples Results: collected and included in this study, 40 were found to be positive for presence of IgM antibodies against Leptospira interrogans. Seropositivity for leptospirosis was observed to be 19%. Maximum number of patients were from economically productive age groups, 31-40 years of age group followed by 21-30 and 41-50 years of age groups. CONCLUSION: Leptospirosis was found to be a major cause of acute febrile illness from north India. It is neglected and under reported from most of the regions of India due to lack of clinician's suspicion. More studies with more samples are required on leptospirosis from this region to reach on final conclusion.

Leptospiral c-di-AMP-binding protein regulates potassium ion transport / 中华微生物学和免疫学杂志

Objective:To investigate whether KtrA was a binding protein of c-di-AMP, the second messenger in Leptospira, and to explore the function and regulatory mechanism of the c-di-AMP-KtrA/B system. Methods:KtrA gene was amplified by PCR and cloned into pET42a plasmid to construct the pET42a ktrA prokaryotic expression vector. Then the vector was transferred into E. coli BL21DE3 to construct an engineering bacterium E. coli BL21DE3 pET42a-ktrA for the expression of recombinant KtrA (rKtrA). The expressed rKtrA was purified by affinity chromatography. BIAcore technology was used to detect the binding ability of rKtrA to c-di-AMP. Bacterial two-hybrid analysis was used to analyze the interaction between KtrA and KtrB in the leptospiral Ktr system with or without exogenetic c-di-AMP. The above genes were then complemented into the potassium transport-deficient E. coli mutants to analyze the function of the c-di-AMP-KtrA/B pathway. Results:An prokaryotic engineering bacterium for the expression of ktrA gene of Leptospira was constructed successfully. The purified rKtrA could specifically bind to c-di-AMP. There was interaction between KtrA and KtrB, but the interaction could be dissociated by c-di-AMP. The KtrA/B system was involved in potassium ion uptake and it was negatively regulated by c-di-AMP. Conclusions:Leptospiral KtrA was a c-di-AMP-binding protein and the c-di-AMP-KtrA/B system was involved in potassium ion transport.

Kidney Injury Biomarkers in Leptospirosis

ABSTRACT Leptospirosis is a zoonotic infection with a global distribution, though it has a greater impact on marginalized rural agricultural and urban communities in developing countries. Kidney injury, which can lead to severe and lethal infections, is the most frequent complication associated with leptospirosis. Novel biomarkers are being studied as tools for assessing kidney injury in different pathological processes to improve early detection. This review aimed to gather information on the use of novel kidney biomarkers for human leptospirosis. A search of the literature was carried out in September 2021 using the parameters "((kidney) OR (renal) OR (chronic kidney disease) OR (acute kidney injury)) AND ((biomarker) OR (marker)) AND ((Leptospira) OR (leptospirosis))". The review identified 11 original studies that evaluated the performance of 15 kidney biomarkers related to leptospirosis. Assessment of the evidence for biomarker utility was limited because of the small number of studies and sample sizes. Although some biomarkers were associated with kidney disease, no specific biomarker appeared to be ready for clinical practice, and more research in this field is necessary.

Seroprevalencia de leptospirosis y brucelosis en búfalos de agua (Bubalus bubalis) en Tierralta Córdoba, Colombia

En la producción zootécnica, la brucelosis y la leptospirosis ocasionan problemas reproductivos y son una limitante en salud y en producción animal. El objetivo fue determinar la presencia de anticuerpos contra Leptospira spp. y Brucella abortus, en una población bufalera, en el municipio de Tierralta, Córdoba. Se realizó un estudio descriptivo de corte transversal, que incluyó un total de 144 búfalos de la raza Murrah, destinados al doble propósito. Para el diagnóstico de Leptospira spp., se implementó la prueba de aglutinación microscópica, con 13 serogrupos, 19 serovares pertenecientes a 5 especies de Leptospira patógenas y para brucelosis Rosa de Bengala y C-Elisa. La seroprevalencia para Leptospira spp. fue del 87,5 %, el serogrupo Mini fue el de mayor frecuencia, pero Grippotyphosa presentó el mayor título. El 16,67 % de los búfalos evaluados presentaron títulos iguales o superiores a 1:800, asociados con infección actual o reciente. La alta seroprevalencia, se puede deber a las características ambientales de la zona, que brinda las condiciones favorables para el crecimiento y el mantenimiento de este patógeno que, sumado al comportamiento de los búfalos de revolcarse, los hace propensos a las infecciones con bacterias del género Leptospira sp., porque a menudo, las fuentes de agua están contaminadas por este patógeno. La seroprevalencia contra B. abortus por Rosa de Bengala y Elisa-C fue del 2,08 %. Todos los títulos determinados corresponden a procesos infecciosos. No hubo signos clínicos de enfermedad y la carencia de registros productivos no permitió determinar el efecto sobre los parámetros reproductivos.

In animal production, brucellosis and leptospirosis cause reproductive problems and limit animal health and production. The objective was to determine the presence of antibodies against Leptospira spp. and Brucella abortus in a buffalo population in the municipality of Tierralta, Cordoba. A descriptive, transversal study was carried out including a population of 144 Murrah´s breed buffalos destined for beef and milk production. For the Leptospira spp. diagnostic, was used a rapid slide agglutination test with 13 serogrups and 19 serovars belonging to 5 pathogenic Leptospira species was implemented and for Brucella abortus Rose Bengal and C-Elisa was used. Seroprevalence of Leptospira spp. was 87,5 %, serogrup Mini was the most frequent, but Grippotyphosa showed the higher titer. The 16,67 % of the buffaloes evaluated presented titles equal to or above than 1:800 associated with current or recent infection. High seroprevalence may be due to environmental characteristics of the zone, which gives favorable conditions for the growth and maintenance of this pathogen, these factors in conjunction with the habit of wallowing makes them prone to suffering infections caused by bacteria of the genera Leptospira sp. since water sources are often contaminated. Seroprevalence against B. abortus by Rose-Bengal and C-Elisa was 2,08 %, the determined titers correspond to infectious processes. There were no clinical manifestations of disease and the effects on reproductive parameters were not determined because of the lack of productive registries.

Molecular detection of leptosipira in synanthropic and wild rodents from Villavicencio municipality, Colombia / Detección molecular de leptospiras en roedores del municipio de Villavicencio, Colombia

Introduction: Rodents are potential transmitters of Leptospira spp. In the municipality of Villavicencio, Colombia, leptospirosis is a disease that, although notifiable, is still underreported. In this region, rodent species that can host pathogenic leptospira remain unknown. Objective: To detect the presence of Leptospira spp. through molecular analysis in rodents (Rodentia) from peri-urban and rural areas belonging to the municipality of Villavicencio in Colombia. Methods: Peri-urban and rural areas of the townships belonging to Villavicencio municipality were selected for sampling. These areas presented similar ecological conditions: they were near water bodies and peridomiciliary areas, and some of them included fields of agricultural crops. Rodents´ kidneys were removed and frozen in liquid nitrogen. DNA was extracted using a commercial kit and subsequently amplified through conventional polymerase chain reaction. Results: The rodent species collected were: Rattus rattus, Mus musculus, Zygodontomys brevicauda, Oligoryzomys sp, Hylaeamys (formerly Oryzomys) and Proechimys cf. oconnelli. Leptospira DNA was amplified in six rodents and the purified amplicons were sent to Macrogen Inc. (Seoul, Korea) for sequencing. The alignment analysis of the sequenced products demonstrated 98.64 percent of coverage and identity with Leptospira interrogans. Conclusions: This is the first study carried out on wild and synanthropic rodents in the municipality of Villavicencio. The incidence of leptospirosis raises the alarm due to the important role of these small mammals in the transmission of this zoonosis, which is considered the second cause, after dengue, of undifferentiated febrile illness in Villavicencio(AU)

Introducción: Los roedores son potenciales transmisores de Leptospira spp. En el municipio de Villavicencio, Colombia, la leptospirosis es una enfermedad que, aunque debe notificarse obligatoriamente, sigue subreportada. En esta región, algunas especies de roedores pueden ser reservorios de leptospiras patógenas, situación que se desconoce. Objetivo: Detectar la presencia de Leptospira spp. a través del análisis molecular en roedores (Rodentia) de áreas periurbanas y rurales del municipio de Villavicencio, Colombia. Métodos: Para el trampeo se seleccionaron áreas periurbanas y rurales de las veredas pertenecientes al municipio de Villavicencio. Las áreas escogidas presentaban condiciones ecológicas similares: cerca de cuerpos de agua y áreas peridomiciliarias; algunas de ellas localizadas en campos de cultivos de la agricultura. Se extirparon los riñones de los roedores y se conservaron en nitrógeno líquido. Se extrajo el ADN usando un estuche comercial y posteriormente se amplificó mediante reacción en cadena de la polimerasa convencional. Resultados: Las especies de roedores colectadas fueron: Rattus rattus, Mus musculus, Zygodontomys brevicauda, Oligoryzomys sp., Hylaeamys (ahora Oryzomys) y Proechimys oconnelli. El ADN de leptospira se amplificó en seis roedores y los amplicones purificados se enviaron a Macrogen Inc. (Seoul, Korea) para secuenciación. El análisis de alineamiento de los productos secuenciados demostró un 98,64 por ciento de cobertura e identidad con Leptospira interrogans. Conclusiones: Este es el primer estudio llevado a cabo en roedores silvestres y sinantrópicos en el municipio de Villavicencio. La incidencia de la leptospirosis genera una alarma con respecto a la importancia del papel de esos pequeños mamíferos en la transmisión de esta zoonosis, la cual es la segunda causa de los síndromes febriles indiferenciados en Villavicencio, después del dengue(AU)

Alteraciones genitales en la infección por leptospiras en ratas Wistar gestadas / Genital alterations in leptospiras infection in pregnant Wistar rats

RESUMEN Introducción: la leptospirosis en Cuba se presenta de forma endémica, y a veces de forma epidémica, por lo que es un problema de salud. Su efecto sobre los genitales internos, en gestantes, ha sido poco tratado por los investigadores. Objetivo: describir las principales alteraciones en los genitales internos de ratas Wistar gestadas infestadas por leptospiras. Métodos: se realizó un estudio experimental en ratas Wistar preñadas: siete grupos con tres animales cada uno: un control negativo y seis casos con dosis de una concentración bacteriana de 300 millones de Leptospiras por mL; se inocularon dos mililitros a cada animal al cuarto día de gestadas por 17 meses. Se tomaron los genitales internos de todas las ratas el día 20 de la gestación, estos fueron procesados en parafina y teñidos con hematoxilina/eosina. Resultados: se observaron alteraciones en útero, ovarios y trompas; el mayor número de lesiones se presentaron en útero mientras que en ovarios y trompas se visualizaron la misma cantidad de lesiones. Conclusiones: la leptospirosis infectó los genitales internos con la producción de quistes, necrosis, abscesos, ooforitis y microhemorragias en ovarios; congestión vascular y hemorragias en las trompas uterinas y endometritis, hemorragias, miometritis, piometristis, perimetritis y congestión vascular en el útero. Predominaron las alteraciones congestivas y hemorrágicas.

ABSTRACT Introduction: leptospirosis in Cuba occurs endemically, and sometimes epidemically, making it a health problem. Its effect on internal genitalia in pregnant women has been little treated by researchers. Objective: to describe the main alterations in the internal genitalia of pregnant Wistar rats infested by leptospiras. Methods: an experimental study was carried out in pregnant Wistar rats: 7 groups with three animals each: 1 negative control and 6 cases with doses of a bacterial concentration of 300 million Leptospira per mL; each animal was inoculated with two milliliters on the fourth day of pregnancy for 17 months. The internal genitalia of all rats were taken on day 20 of gestation, processed in kerosene and stained with hematoxylin/eosin. Results: alterations were observed in the uterus, ovaries and tubes; the greatest number of lesions was present in the uterus while the same number of lesions was visualized in the ovaries and tubes. Conclusions: leptospirosis infected the internal genitalia producing cysts, necrosis, abscesses, oophoritis and microhemorrhages in the ovaries; vascular congestion and hemorrhages in the uterine tubes and endometritis, hemorrhages, myometritis, pyometristis, perimetritis and vascular congestion in the uterus. Congestive and hemorrhagic alterations predominated.

Improvement of the enrichment used in the EMJH médium (Ellinghausen-McCullough-Johnson-Harris) for the cultivation of Leptospira spp

Abstract Changes were made to the original formulation of the EMJH medium (Ellinghausen-McCullough-Johnson-Harris) enrichment and some aspects such as growth time of Leptospira and utilization in the microscopic agglutination test (MAT) were evaluated and compared to the original enrichment and to a commercially available enrichment (DIFCOÔ). Leptospira samples (24 antigens) that make up our panel of antigens used in MAT were used, among them, reference and autochthonous strains isolated in Brazil. The samples were grown individually in the EMJH medium under the three previously mentioned conditions (adapted enrichment, original enrichment and commercial enrichment). In addition, 89 blood serums from domestic and wild animals were analyzed by MAT using the antigens grown in these media. All samples tested grew efficiently with the adapted enrichment, and the MAT results were satisfactory. Therefore, other laboratories could also benefit from the use of this adapted enrichment when culturing the Leptospira strains applied in their MAT panels.

Resumen Realizamos cambios en la formulación original del enriquecimiento del medio Ellinghausen-McCullough-Johnson-Harris y evaluamos algunos aspectos, como el tiempo de crecimiento de Leptospira y la utilización en la prueba de aglutinación microscópica (MAT), comparándolos con el enriquecimiento original y un medio de enriquecimiento comercialmente disponible (DIFCOÔ). Se usaron muestras de Leptospira (24 antígenos) que componen nuestro panel de antígenos utilizados en la MAT, entre ellos, cepas autóctonas y de referencia aisladas en Brasil. Las muestras se cultivaron individualmente en medio Ellinghausen-McCullough-Johnson-Harris en las tres condiciones mencionadas (enriquecimiento adaptado, enriquecimiento original y enriquecimiento comercial). Adicionalmente, 89 sueros de sangre de animales domésticos y salvajes fueron analizados por MAT usando los antígenos cultivados en estos medios. Todas las muestras analizadas crecieron eficientemente con el enriquecimiento adaptado y los resultados de la MAT fueron satisfactorios. Por lo tanto, otros laboratorios también podrían beneficiarse del uso de este enriquecimiento adaptado al cultivar las cepas de Leptospira aplicadas en sus paneles MAT.

Tendência temporal da leptospirose e sua associação com variáveis climáticas e ambientais em Santa Catarina, Brasil / Temporal trends in leptospirosis incidence and association with climatic and environmental factors in the state of Santa Catarina, Brazil

Resumo A leptospirose é uma zoonose que apresenta potencial epidêmico, principalmente após fortes chuvas que acarretam inundações, alagamentos e enxurradas. Algumas características da região costeira de Santa Catarina, localizada no Sul do Brasil, influenciam nesses processos. Portanto, a partir do estudo da leptospirose nos seis municípios do estado com as maiores incidências e picos epidêmicos de 2000 a 2015, buscou-se conhecer a tendência dessa doença e as variáveis climáticas e ambientais associadas à sua ocorrência, ajustando dois modelos com resposta binomial negativa. As maiores incidências foram encontradas em 2008 e 2011, com picos no mesmo mês ou no posterior aos eventos de desastres. A incidência apresentou forte comportamento sazonal, sendo maior nos meses do verão. Observou-se tendência de queda na incidência dos municípios estudados, estimada em 3,21% ao ano. Os fatores climáticos e ambientais mais fortemente associados foram o número de dias de chuva, a temperatura máxima e a presença de enxurrada e de inundação, com diferentes impactos entre os municípios. Houve interações significativas, indicando que o efeito de inundações na incidência não é o mesmo em todos os municípios e que as diferenças nas incidências entre os municípios dependem da ocorrência ou não de inundações.

Abstract Leptospirosis is a zoonosis with epidemic potential, especially after heavy rainfall causing river, urban and flash floods. Certain features of Santa Catarina's coastal region influence these processes. Using negative binomial regression, we investigated trends in the incidence of leptospirosis in the six municipalities with the highest epidemic peaks between 2000 and 2015 and the climatic and environmental variables associated with the occurrence of the disease. Incidence was highest in 2008 and 2011, and peaks occurred in the same month or month after disasters. Incidence showed a strong seasonal trend, being higher in summer months. There was a decrease trend in incidence across the six municipalities (3.21% per year). The climatic and environmental factors that showed the strongest associations were number of rainy days, maximum temperature, presence of flash floods, and river flooding. The impact of these variables varied across the municipalities. Significant interactions were found, indicating that the effect of river flooding on incidence is not the same across all municipalities and differences in incidence between municipalities depend on the occurrence of river flooding.

Daños renales en fetos de ratas Wistar infectadas con Leptospira canicola

RESUMEN Introducción: La leptospirosis es una de las zoonosis que en Cuba se presenta de forma endémica, es causante de pérdidas humanas, animales y económicas, por lo que es un problema de salud. Las ratas constituyen un vector transmisor de gran importancia. Trabajos experimentales para estudiar las alteraciones renales fetales en modelo de rata Wistar no se han realizado en Cuba. Objetivo: Identificar las principales alteraciones morfométricas renales en fetos de ratas Wistar infectadas con serovariedad Leptospira canicola, en la Universidad de Ciencias Médicas de Pinar del Rio en el período comprendido entre 2017-2019. Método: Se realizó un estudio experimental en ratas Wistar preñadas: 2 grupos con tres animales cada uno: el grupo experimental con dosis de una concentración bacteriana de 300 millones de leptospiras por mililitros; se inoculó 1 ml a cada una infectándolas con cepas de Leptospira canicola. Se estudiaron los riñones de fetos extraídos por cesárea y se analizaron las variables: longitud renal y peso del riñón. Resultados: Los resultados obtenidos confirmaron los daños renales producidos por la Leptospira canicola tanto en el eje mayor como el menor de ambos riñones, así como variaciones en el peso de las crías infectadas. Conclusiones: La infección por Leptospira canicola en ratas Wistar preñadas provoca alteraciones en los riñones de los fetos lo que causa cambios en la morfometría de los ejes tanto mayor como menor, así como en el peso corporal del feto, lo que conlleva a una disminución del peso renal.

ABSTRACT Introduction: Leptospirosis is one of the endemic zoonoses in Cuba, causing human, animal and economic losses, making it a significant health problem. Rats are a very remarkable vector. Experimental work to study fetal renal alterations in the Wistar rats has not been carried out in Cuba. Objective: To identify the main renal morphometric alterations in fetuses of Wistar rats infected with serovar Leptospira canicola, at the Universidad de Ciencias Médicas de Pinar del Rio in the period between 2017-2019. Method: An experimental study was carried out in pregnant Wistar rats: 2 groups with three animals each: the experimental group with a dose of bacterial concentration of 300 million leptospires per milliliter; 1 ml was inoculated to each one, infecting them with strains of Leptospira canicola. The kidneys of fetuses extracted by cesarean section were studied, and the variables: renal length and kidney weight were analyzed. Results: The results obtained confirmed kidney damage caused by Leptospira canicola both in the major and minor axis of both kidneys, as well as variations in the weight of the infected pups. Conclusions: Leptospiral infection in pregnant Wistar rats causes alterations in the kidneys of the fetuses, bringing changes in the morphometry of both the major and minor axes, as well as in the body weight of the fetus, which leads to a decrease in renal weight.

RESUMO Introdução: A leptospirose é uma das zoonoses endêmicas em Cuba, causando prejuízos humanos, animais e econômicos, tornando-se um problema de saúde. Os ratos são um vetor transmissor muito importante. Trabalho experimental para estudar as alterações renais fetais no modelo de rato Wistar não foi realizado em Cuba. Objetivo: Identificar as principais alterações morfométricas renais em fetos de ratos Wistar infectados com sorovar Leptospira canicola, na Universidad de Ciencias Médicas de Pinar del Rio no período entre 2017-2019. Método: Estudo experimental realizado em ratas Wistar prenhes: 2 grupos com três animais cada: o grupo experimental com dose de concentração bacteriana de 300 milhões de leptospiras por mililitro; 1 ml fora inoculado em cada um, infectando-os com cepas de Leptospira canicola. Foram estudados os rins de fetos extraídos por cesariana e analisadas as variáveis: comprimento renal e peso renal. Resultados: Os resultados obtidos confirmaram o dano renal causado por Leptospira canicola tanto no eixo maior quanto no eixo menor de ambos os rins, bem como variações no peso dos filhotes infectados. Conclusões: A infecção por Leptospira canicola em ratas Wistar prenhes causa alterações nos rins dos fetos, o que provoca alterações na morfometria dos eixos maior e menor, bem como no peso corporal do feto, o que leva a uma diminuição da peso do rim.

Detection of Pathogenic Leptospira in Rats and Phylogenetic Analysis Using Outer Membrane Lipoprotein Lipl32 Gene at Two Major Public Markets

@#Introduction: Limited studies have been documented on the presence of pathogenic Leptospira in public markets serving the community in sub-districts of Selangor. The aim of this study was to detect the presence of pathogenic Leptospira in rats using a gene encoding an outer membrane lipoprotein LipL32. Methods: Polymerase chain reaction (PCR) was performed using LipL32 primers on sixty kidney samples of rats trapped at two locations of study; Pasar Borong Selangor in Seri Kembangan and Pasar Basah Bandar Baru Bangi in Bangi. Results: Out of 60 samples analysed, 36.7% were positive for the presence of LipL32. All positive samples highly matched (>94%) nucleotide sequence for LipL32 of pathogenic Leptospira and related to the pathogens through phylogenetic analysis. Conclusion: The detection of LipL32 indicates the potential presence of pathogenic Leptospira species at public markets. Although only 60 rats were successfully trapped, the rats are mobile and might further transmit the pathogenic organisms to other areas.

Primeros pasos en la inmunoprofilaxis contra la leptospirosis en Cuba

RESUMEN Introducción: La leptospirosis es una zoonosis de distribución mundial. En la década de los años 70 del pasado siglo xx aumentó la incidencia en Cuba, principalmente en trabajadores agrícolas. En Camagüey se produjo un brote que llevó a establecer el Programa Nacional de Control de la Leptospirosis Humana, puesto en vigor por los Ministerios de Salud Pública y la Agricultura. En 1982 investigadores de la Dirección Nacional de Zoonosis realizaron un ensayo clínico para evaluar inmunogenicidad y reactogenicidad de una vacuna antileptospirósica humana del Instituto de Sueros y Vacunas de Stávropol. Objetivo: Reconstruir los primeros pasos en la inmunoprofilaxis contra la leptospirosis en Cuba. Métodos: Se utilizó el método historiográfico. Los datos se obtuvieron en las bases de datos: Infomed, Dialnet, Medigraphic, Medline y Scielo. Análisis y síntesis de la información: Las recomendaciones de la investigación plantearon aplicar la vacuna a grupos de riesgo y producir una vacuna con cepas autóctonas de leptospiras. El análisis de la bibliografía consultada de las últimas cuatro décadas corrobora el cumplimiento de las recomendaciones. Conclusiones: Hace cuarenta años científicos cubanos realizaron los estudios iniciales sobre inmunogenicidad y reactogenicidad de una vacuna procedente de la pretérita Unión Soviética. Las recomendaciones de la investigación se cumplieron. Se realizó la vacunación a grupos de riesgo en la población, principalmente trabajadores agrícolas y se logró una vacuna con cepas autóctonas de leptospiras. Los datos expuestos sobre los primeros pasos en la inmunoprofilaxis contra la leptospirosis en Cuba tienen relevancia para la historiografía médica por los aportes a la salud pública, prioridad del Estado cubano.

ABSTRACT Introduction: Leptospirosis is a zoonosis of worldwide distribution. In the 1970s, there was an increase in the incidence of this disease in Cuba, mainly among agricultural workers. An outbreak in Camagüey led to the establishment of the National Program for the Control of Human Leptospirosis adopted by the Ministries of Public Health and Agricultures. In 1982, a clinical trial was conducted to evaluate the immunogenicity and the reactogenicity of a vaccine against human leptospirosis from the Institute of Vaccines and Sera of Stavropol. Objective: To retrace the first steps towards immunoprophylaxis against leptospirosis in Cuba. Methods: The historiographic method was used. Data were retrieved from the following databases: Infomed, Dialnet, Medigraphic, Medline and SciELO. Analysis and synthesis of the information: Recommendations from the research suggested administering the vaccine to risk groups, and producing a vaccine from autochthonous Leptospira strains. The analysis of the consulted bibliography from the last four decades corroborates the fulfillment of these recommendations. Conclusions: Forty years ago, Cuban scientists conducted the first studies on immunogenicity and reactogenicity of a vaccine from the former Soviet Union. The recommendations derived from the research were fulfilled. Population groups at risk, mainly agricultural workers, were vaccinated, and a vaccine from autochthonous Leptospira strains was obtained. Given the contributions to public health, which is a priority of the Cuban government, data presented on the first steps towards immunoprophylaxis against leptospirosis in Cuba are of paramount importance for the medical historiography.

Caracterização funcional de proteínas hipotéticas de Leptospira interrogans como adesinas e potenciais antígenos vacinais e para diagnóstico / Functional characterization of hypothetical proteins in Leptospira interrogans as adhesins and potential vaccine and diagnostic antigens

Leptospira spp. constitui um grupo de bactérias espiroquetas gram-negativas englobando espécies saprofíticas, intermediárias e patogênicas, sendo as últimas agentes causadores da leptospirose, doença zoonótica de alcance mundial e endêmica em regiões tropicais em desenvolvimento. O crescente número de espécies identificadas de leptospiras destaca ainda mais sua diversidade genética e mecanismos de virulência únicos, muitos deles com função ainda desconhecida. Esforços para o desenvolvimento de novas vacinas com proteção cruzada e efeito duradouro revelaram possíveis candidatos vacinais que necessitam ser adequadamente validados, sendo assim, há ainda uma urgente necessidade de uma vacina universal contra a leptospirose capaz de controlar e reduzir os surtos cada vez mais frequentes da doença. Adesinas são importantes fatores de virulência em diversos patógenos, constituindo antígenos promissores para o desenvolvimento de vacinas contra a leptospirose, assim como para o desenvolvimento de métodos diagnósticos mais rápidos e precisos. Previamente, foram identificadas três proteínas hipotéticas conservadas em L. interrogans pela técnica de phage display, denominadas arbitrariamente como LepA069, LepA962 e LepA388. A expressão do gene codificador da proteína LepA069 apresentou aumento de aproximadamente 70 % em animais infectados por leptospiras virulentas, representando a primeira evidência funcional desta proteína ainda desconhecida. Porções recombinantes da lipoproteína hipotética LepA962 (LepA962_Nt e LepA962_Phg) foram obtidos, sendo demonstrada a forte interação da proteína LepA962_Phg, contendo a sequência identificada por phage display, com laminina, fibronectina plasmática, colágeno I e fibrinogênio de maneira dose-dependente. Adicionalmente, LepA962_Phg apresentou ligação às células VERO e à sua matriz extracelular secretada, e o soro obtido a partir desta proteína recombinante foi capaz de se ligar à superfície de leptospiras virulentas, indicando que LepA962_Phg pode representar um importante domínio de interação entre as leptospiras e seu hospedeiro. Finalmente, a proteína LepA388 pertencente a uma extensa família de proteínas modificadoras de virulência com função desconhecida (DUF_61), presente apenas nas leptospiras patogênicas mais virulentas, apresentou aumento na expressão de seu gene codificador em animais infectados por leptospiras virulentas de acordo com dados na literatura. Além disso, porções recombinantes da região Nterminal desta proteína apresentaram ligação a laminina, colágenos I e IV, vitronectina e fibronectinas plasmática e celular, principalmente considerando a sequência identificada por phage display. Estes dados reforçam as predições de modelos tridimensionais da proteína LepA388 e de outros membros da família DUF_61, as quais identificam domínios semelhantes a toxinas (como abrina e CARDS) responsáveis pela ligação e internalização celulares nos hospedeiros. Dados recentes sugerem um possível papel citotóxico desempenhado pelas proteínas desta família em leptospiras, as quais podem também ser consideradas potenciais candidatas vacinais e para diagnóstico da leptospirose, devido à sua distribuição restrita em espécies e cepas patogênicas de importância para saúde humana.

Leptospira spp. constitutes a group of gram-negative spirochete bacteria comprising saprophytic, intermediate and pathogenic species, the last being causative agents of leptospirosis, a zoonotic disease of worldwide extent and endemic in developing tropical regions. The growing number of identified leptospiral species further highlights their genetic diversity and unique virulence mechanisms, many of them with unknown function. Efforts to develop new vaccines with cross-protection and long-lasting effect have revealed possible vaccine candidates that need to be properly validated. Therefore, there is still an urgent need for a universal vaccine against leptospirosis capable of controlling and reducing the increasing outbreaks of the disease. Adhesins are important virulence factors in several pathogens, constituting promising antigens for the development of vaccines against leptospirosis, as well as for the development of faster and more accurate diagnostic methods. Previously, three conserved hypothetical proteins in L. interrogans were identified by phage display technique, arbitrarily named as LepA069, LepA962 and LepA388. Expression of the LepA069 encoding gene showed an increase of approximately 70 % in animals infected by virulent leptospires, representing the first functional evidence of this still unknown protein. Recombinant portions of the hypothetical lipoprotein LepA962 (LepA962_Nt and LepA962_Phg) were obtained, demonstrating the strong interaction of the LepA962_Phg protein, containing the sequence identified by phage display, with laminin, plasma fibronectin, collagen I and fibrinogen in a dose-dependent manner. Furthermore, LepA962_Phg showed binding to VERO cells and its secreted extracellular matrix, and the serum obtained from this recombinant protein was able to bind to the surface of virulent leptospires, indicating that LepA962_Phg may represent an important domain of interaction between leptospires and its host. Finally, LepA388 protein belonging to an extensive family of virulence modifying proteins with unknown function (DUF_61), present only in the most virulent pathogenic leptospires, showed an increase in the expression of its encoding gene in animals infected by virulent leptospires according to data in literature. Moreover, recombinant portions of the N-terminal region of this protein showed binding to laminin, collagens I and IV, vitronectin and plasma and cell fibronectins, especially considering the sequence identified by phage display. These data support the predictions of three-dimensional models of the LepA388 protein and other members of the DUF_61 family, which identify toxin-like domains (such as abrin and CARDS) responsible for cellular binding and internalization in hosts. Recent data suggest a possible cytotoxic role played by proteins of this family in leptospires, which can also be considered potential vaccine candidates and antigens for diagnosis, due to their restricted distribution in pathogenic species and strains of importance to human health

Factores ecoambientales relacionados con potenciales reservorios de Leptospira spp. en la Reserva Ecológica Costanera Sur, Buenos Aires, Argentina / Eco-environmental Factors Related to Potential Reservoirs of Leptospira spp. in the Costanera Sur Ecological Reserve, Buenos Aires, Argentina

Resumen Las modificaciones ambientales generan entornos de interfaz en los que la transmisión de Leptospira spp. puede verse afectada. Con el objetivo de identificar y analizar variables ecoambientales asociadas a potenciales reservorios de Leptospira spp. en la Reserva Ecológica Costanera Sur (RECS) (Buenos Aires, Argentina) y áreas urbanizadas contiguas, se realizó un muestreo por conveniencia en líneas transectas, estratificado y proporcionado. Fueron capturados 170 mamíferos de 11 especies y se analizó la presencia de Leptospira spp. por serología, cultivo y PCR. En áreas conservadas, se registraron mayormente Didelphis albiventris y Deltamys kempi, mientras que Oligoryzomys flavescens también fue hallado en áreas degradadas, y Rattus norvegicus solo fue encontrada en áreas degradadas. No se detectó Leptospira spp. en los mamíferos silvestres. Un perro sin tenedor responsable resultó positivo por serología. Las condiciones ambientales de la RECS sugieren un ecosistema poco propicio para Rattus, evidenciando la importancia de la conservación de áreas naturales en entornos urbanos.

Abstract Environmental modifications generate interface environments in which Leptospira spp. transmission may be affected. In order to identify and analyze eco-environmental variables associated with potential reservoirs of Leptospira spp. in the Reserva Ecológica Costanera Sur (RECS) (Buenos Aires, Argentina) and adjacent urbanized areas, a stratified and proportionate transect line convenience sampling was carried out. A total of 170 mammals of 11 species were captured and analyzed for the presence of Leptospira spp. by serology, culture and PCR. In conserved areas, Didelphis albiventris and Deltamys kempi were mostly recorded, while Oligoryzomys flavescens was also found in degraded areas, and Rattus norvegicus was only found in degraded areas. Leptospira spp. were not detected in wild mammals. One dog without a responsible keeper tested positive by serology. The environmental conditions of the RECS suggest a poor ecosystem for Rattus, demonstrating the importance of the conservation of natural areas in urban environments.

Estudo retrospectivo da leptospirose canina em hospital veterinário escola no município de Londrina, Pr / Retrospective study of canine leptospirosis in a veterinary school hospital in the municipality of Londrina, Pr / Estudio retrospectivo de leptospirosis canina en un hospital veterinario escolar del municipio de Londrina, Pr

O objetivo do trabalho foi relatar a ocorrência de casos de cães com Leptospira spp. no Hospital Veterinário de Londrina. Foram avaliados 5.454 prontuários, durante os meses de janeiro a dezembro de 2016, e selecionados os que apresentaram solicitação de exame para Lepstopira por meio da técnica de campo escuro (CE). Nos positivos pela técnica CE dados sobre o sexo, sinais clínicos, endereço, data do atendimento e resultado na SAM foram coletados. A análise dos dados foi realizada pelo programa EpiInfo (7.2.3.1), dos 5.454 prontuários nos quais foi solicitado exame CE em 7,06% (385/5.454), e nesses, 13,76% (53/385) foi identificada a presença da espiroqueta. Dos animais positivos na técnica CE, 20,75% (11/53) foram reagentes na SAM. O sorovar Canicola foi o mais frequente em 90,91% (10/11) dos animais, apresentando títulos variados de 100 a 12.800. Para o sorovar Pomona 9,09% (1/11) dos animais apresentaram títulação de 100. Os sinais clínicos mais observados foram apatia em 58,49% (31/53), azotemia em 49,06% (26/53) e êmese em 49,06% (26/53) dos casos. Quanto ao sexo, os machos constituíram 71,70% dos casos e as fêmeas 28,30% (p=0,0015). Os resultados demonstraram a presença de Leptospira spp. entre os animais atendidos no hospital veterinário de Universidade Estadual de Londrina e reforçam a importância do sorovar Canicola entre os cães, visto que esse foi o mais frequente entre os sorovares e que apresentou os maiores títulos.(AU)

The purpose of this work is to report the occurrence of cases of dogs with Leptospira spp. at the Veterinary Hospital of Londrina. A total of 5,454 medical records were evaluated from January to December 2016, and data obtained from the examination for Lepstopira using the dark field technique (DF) were selected. In cases which were positive by the DF technique, data on gender, clinical signs, address, care data, and SAM results were collected. Data analysis was performed using the EpiInfo program (7.2.3.1). From the total 5,454 medical records in which DF examination was requested, in 7.06% (385/5.454), and from those, 13.76% (53/385) were positive for Leptospira. From the animals positive in the DF technique, 20.75% (11/53) were reactive in SAM. The Canicola serovar was the most prevalent, being present in 90.91% (10/11) of the animals, with titers ranging from 100 to 12,800. For the Pomona serovar, 9.09% (1/11) of the animals presented a title of 100. The most frequent clinical signs observed were apathy, in 58.49% (31/53); azotemia in 49.06% (26/53); and emesis in 49.06% (26/53) of the cases. As for gender, male animals constituted 71.70% of the cases and female 28.30% (p = 0.0015). The results demonstrated the presence of Leptospira spp. among animals treated at the Veterinary Hospital of the State University of Londrina which reinforces the importance of the Canicola serovar among dogs, since it was the serovar with the highest prevalence and highest titer found.(AU)

El objetivo de este trabajo fue reportar la ocurrencia de casos de perros con Leptospira spp. en el Hospital Veterinario de Londrina. Se evaluaron un total de 5.454 historias clínicas de enero a diciembre de 2016, y se seleccionó los que presentaron solicitud de examen de Lepstopira mediante la técnica de campo oscuro (CE). En los positivos por la técnica de CE se recogieron datos de sexo, signos clínicos, domicilio, fecha de atención y resultado en la SAM. El análisis de los datos se realizó mediante el programa EpiInfo (7.2.3.1), de las 5.454 historias clínicas en las que se solicitó examen CE en el 7,06% (385/5.454), y en estas, el 13,76% (53/385) se identificaron la de espiroqueta. De los animales positivos en la técnica CE, el 20,75% (11/53) fueron reactivos en la SAM. El serovar Canicola fue el más frecuente en el 90,91% (10/11) de los animales, con títulos que oscilaron entre 100 y 12.800. Para el serovar Pomona, el 9.09% (1/11) de los animales tuvo un título de 100. Los signos clínicos más observados fueron apatía en 58.49% (31/53), azotemia en 49.06% (26/53) y emesis en 49.06% (26/53) de los casos. Cuanto al sexo, los machos constituyeron el 71,70% de los casos y las hembras el 28,30% (p = 0,0015). Los resultados demostraron la presencia de Leptospira spp. entre los animales tratados en el Hospital Veterinario de la Universidad Estatal de Londrina y refuerzan la importancia del serovar Canicola entre los perros, ya que este fue el más frecuente entre los serovares y tuvo los títulos más altos.(AU)

First report of pathogenic Leptospira spp. in Tadaridabrasiliensis bats (family Molossidae) and Eptesicus furinalis (family Vespertilionidae) of Argentina. New host species in this country? / Primer reporte de Leptospira spp. patógena en murciélagos Tadaridabrasiliensis (familia: Molossidae) y Eptesicusfurinalis (familia: Vespertilionidae) en Argentina. ¿Nuevas especies hospedadoras en el país?

ABSTRACT Leptospirosis is an endemic disease caused byLeptospiraspp., a bacterium that affects animals and humans. In recent years, the number of reports of leptospirosis in wild animals has increased, which highlights the need to study the infectious agents in these animals. In this study, a duplex PCR for the detection of leptospiral DNA was performed on 50 kidney samples from bats, and a MAT (Microscopic Agglutination Test) for serological detection of anti-leptospiral antibodies was applied to 47 serum samples from bats from different regions of Buenos Aires Province, Argentina. DNA was extracted using Chelex-100 and duplex PCR was performed by targeting the detection of genessecYandflaB, of pathogenicLeptospiraspp. Of the 50 kidney samples, 3 were positive forEumopssp. andTadaridabrasiliensisby duplex PCR. Of the 47 serum samples, 12 were positive for different serovars:Leptospira interrogansserovars Icterohaemorrhagiae, Cynopteri and Bataviae, andLeptospira borgpeterseniiserovar Ballum. This is the first report of the detection of pathogenic leptospires by serology in bats belonging to theT. brasiliensisandEptesicus furinalisspecies in Argentina. In addition, this is the first report of the detection of pathogenic leptospiral DNA by PCR inT. brasiliensisspecies. The detection ofLeptospiraspp. in these wild animals shows that they may play an important role as wildlife reservoirs of leptospires.

RESUMEN La leptospirosis es una enfermedad endémica causada porLeptospiraspp., una bacteria que afecta a animales y a humanos. En los últimos años, el número de reportes de leptospirosis en animales silvestres ha aumentado, lo que resalta la necesidad de analizar los agentes infecciosos en estos animales. En este estudio, se aplicó una reacción en cadena de la polimerasa (PCR) dúplex para la identificación del ADN leptospiral en 50 muestras de riñones de murciélagos y la prueba de aglutinación microscópica (MAT) para la detección serológica de anticuerpos antileptospira en 47 muestras de suero de murciélagos de diferentes regiones de la provincia de Buenos Aires, Argentina. El ADN fue extraído usando Chelex-100 y la PCR dúplex estuvo dirigida a la detección de los genessecYyflaBdeLeptospiraspp. patógena. De las 50 muestras de riñón, tres resultaron positivas por PCR dúplex paraEumopssp. yTadaridabrasiliensis. De las 47 muestras de suero, 12 fueron positivas a diferentes serovares:LeptospirainterrogansserovaresIcterohaemorrhagiae, Cynopteri y Bataviae, yLeptospiraborgpeterseniiserovarBallum. Este es el primer reporte de detección de leptospiras patógenas por serología en murciélagos pertenecientes a las especiesT. brasiliensisyEptesicusfurinalisen Argentina. Además, también es el primero en la localización de ADN leptospiral por PCR en la especieT. brasiliensis.La identificación deLeptospiraspp. en estos animales silvestres muestra que pueden desempeñar un papel importante como reservorios de leptospiras en la fauna silvestre.

A molecular survey reveals high occurrence of co-infections in intensive pork production farms with increased rates of mummified swine fetuses in Southern Brazil / [Pesquisa molecular revela alta ocorrência de coinfecções em granjas de produção intensiva de suínos com altas taxas de mumificação fetal no sul do Brasil]

Neste estudo, 308 amostras de fetos mumificados foram testadas para parvovírus suíno (PPV), circovírus suíno tipos 2 e 3 (PCV2 e PCV3) e leptospiras patogênicas. A idade gestacional no momento da perda gestacional e a frequência da mumificação fetal de acordo com a ordem de parto também foram investigadas. As amostras foram coletadas em granjas comerciais de criação de suínos da região sul do Brasil que apresentassem taxas de mumificação fetal igual ou maiores a 2,5%. Fragmentos de pulmão, rim, fígado e coração de fetos suínos mumificados foram coletados para análise molecular. Resultados da PCR foram classificados de acordo com a região de origem das amostras, tendo Santa Catarina, Paraná e Rio Grande do Sul contabilizado 87 (28,25%), 89 (28,90%) e 132 (42,86%) do total de amostras de fetos suínos mumificados, respectivamente. Coinfecções foram observadas na maioria dos casos e PCV3 foi o agente mais prevalente detectado, encontrado em 298 amostras (96,75%). A maioria das perdas gestacionais foi observada entre 50 e 70 dias de gestação (168; 54,5%) e a mumificação fetal não foi associada à ordem de parto das matrizes. Os achados sugerem que as altas taxas de fetos suínos mumificados na região Sul do Brasil podem ser explicadas pela infecção com esses agentes virais.(AU)

Glucocorticoides en hemorragia alveolar por leptospira / Glucocorticoids in leptospira alveolar hemorrhage

Resumen Se presentan los casos clínicos de dos pacientes que ingresaron en la guardia de Emergencias del HIGA San Martín de La Plata con un cuadro clínico compatible con leptospirosis, que evolucionaron con insuficiencia respiratoria y hemorragia alveolar. En ambos se administraron glucocorticoides con buena evolución. Se realizó una búsqueda bibliográfica de artículos publicados desde 2005 en castellano e inglés y la revisión del tema. Basada en la evidencia actual no se puede hacer una clara recomendación para el uso de corticoides en la leptospirosis grave. La bibliografía publicada es escasa y de baja calidad. Aparentemente habría un beneficio en el uso de corticoides en los casos de afectación pulmonar por leptospirosis grave. Se necesitan estudios de alta calidad para realizar recomendaciones con evidencia científica, para verificar la dosis adecuada de corticoides, tiempo de inicio, duración del tratamiento y los casos en los que se debería administrar esta terapéutica.

Abstract We present the clinical cases of two patients who were admitted to the HIGA San Martín de La Plata emergency ward with a clinical picture compatible with leptospirosis, who evolved with respiratory failure and alveolar hemorrhage. In both, glucocorticoids were administered with good evolution. A bibliographic search of articles published since 2005 in Spanish and English and a review of the topic was carried out. Based on the current evidence, no clear recommendation can be made for the use of corticosteroids in severe leptospirosis. The published bibliography is scarce and of low quality. There would appear to be a benefit in the use of corticosteroids in cases of pulmonary involvement due to severe leptospirosis. High-quality studies are needed to make recommendations with scientific evidence, to verify the adequate dose of corticosteroids, time of initiation, duration of treatment and the cases in which this therapy should be administered.

Monitoring of Leptospira in captive turtles by DNA analysis / [Monitoramento de Leptospira em quelônios mantidos em cativeiro pela análise de DNA]

Existem poucos estudos sobre doenças infecciosas em animais silvestres. O objetivo deste estudo foi pesquisar DNA de Leptospira spp. em sangue de tartarugas mantidas em cativeiro, pertencentes ao Bosque Rodrigues Alves (Jardim Zoobotânico da Amazônia). O DNA foi isolado das amostras de sangue coletadas de 148 tartarugas pertencentes a seis espécies diferentes. A reação em cadeia da polimerase (PCR) foi realizada utilizando-se iniciadores específicos para DNA de Leptospira spp. Nenhuma das amostras apresentou resultado positivo para Leptospira spp.(AU)

Distribution of pathogenic Leptospira in environmental water and soils of selected recreational forests in Perak, Malaysia

@#Leptospirosis is an emerging zoonotic disease endemic in tropical regions. Aiming at assessing the potential infection risks via recreational exposure, the molecular prevalence of pathogenic Leptospira in 14 amenity forests in five selected districts of the state of Perak was determined. Water and soil samples along streams and waterfalls were subjected to culture of leptospires and the pathogenic Leptospira spp. was detected by lipL32-based polymerase chain reaction (PCR). Twenty out of 154 samples (13%) that tested positive for leptospires were mostly soils and still water recorded with tolerable temperatures (22.226.5°C) and pHs (5.73-6.70). The localised prevalence was highly varied among eight positive forests (6.7-41.7%), particularly higher in Kampar and Kinta districts which are the more populated urban areas. The importance of public health surveillance should not be underrated given the high prevalence of Leptospira spp. in forests in close proximity to indigenous settlements, even where the places are clean. Overall, this study discovered a wide distribution of pathogenic Leptospira spp. in recreational areas.