Presence of Leptospira spp. in a Mosaic of Wetlands Used for Livestock Raising under Differing Hydroclimatic Conditions.

Knowledge about the life cycle and survival mechanisms of leptospires in the environment is scarce, particularly regarding the environmental factors associated with their presence in ecosystems subject to livestock farming, where precipitation, seasonal floods, and river overflows could act as facilitators of leptospire dispersion. This study aimed to identify and study the presence of Leptospira spp. in the Lower Delta of the Paraná River and describe the physical, chemical, and hydrometeorological conditions associated with their presence in wetland ecosystems impaired by livestock raising intensification. Here, we show that the presence of Leptospira was determined mainly by water availability. We detected the species Leptospira kmetyi, L. mayottensis, and L. fainei and successfully cultured the saprophytic species L. meyeri from bottom sediment, suggesting the association of leptospires with microbial communities of the sediment's biofilm to enhance its survival and persistence in aquatic environments and adapt to changing environmental conditions. Knowledge of Leptospira sp. diversity in wetlands and the impact of climate variability on the transmission of these organisms is crucial for predicting and preventing leptospirosis outbreaks in the context of human health. IMPORTANCE Wetlands are environments that are often conducive to the survival and transmission of Leptospira because they provide a suitable habitat for the bacteria and are often home to many animal species that can act as reservoirs for leptospirosis. Bringing humans and animals into closer contact with contaminated water and soil and increased frequency and intensity of extreme weather events may further exacerbate the risk of leptospirosis outbreaks, which is mostly relevant in the context of climate change and a widespread intensification of productive activities, particularly in the Lower Delta of the Paraná River. The detection of leptospiral species in wetland ecosystems impaired by livestock raising intensification can help to identify propitious environmental factors and potential sources of infection, develop preventive measures, and plan for appropriate responses to outbreaks, ultimately improving public health outcomes.

Utility evaluation of two molecular methods for Leptospira spp. typing in human serum samples.

Most of the available genotyping methods were applied and evaluated in Leptospira isolates and only few of them in a relevant sample size of blood specimens but not of sera. The objective of this study was to evaluate the utility of one partial 16S rRNA gene sequencing assay (16S rRNA) and an optimized. Multilocus sequence typing scheme (MLST) for Leptospira typing directly in serum samples. Confirmed leptospirosis patients (n = 228) from Argentina (2005-2016) were randomly included. Septicemic-phase serum samples (n = 228) were studied by two genotyping methods. Available immune-phase serum samples of the included patients (n = 159) were studied by MAT to compare serological and molecular results. In culture-proven cases (n = 8), genotyping results between clinical samples and isolates were compared. Typing success rate (TSR) was 21.9% for 16S rRNA and 11.4% for MLST (full allelic profile) and a positive trend in both TSR during the study period was observed. Two species (L. interrogans and L. borgpertesenii) were identified by both methods and MLST assigned 8 different STs. The probable serogroups identified by MLST were coincident with the presumptive infecting serogroups identified by MAT, but with different frequencies. The three serogroups (Canicola, Sejroe and Icterohaemorrhagiae) most frequently identified by MAT were also genotyped by MLST. Typing results via 16S rRNA and MLST in clinical samples and isolates of culture-proven cases, were consistent except for one case. Performance of partial 16S rRNA gene sequencing assay and the optimized MLST scheme directly in sera may increase and improve the knowledge about species and serogroups causing human leptospirosis, especially in countries with low rates of culture sample collection or Leptospira isolation.

Determining the spatial distribution of environmental and socio-economic suitability for human leptospirosis in the face of limited epidemiological data.

BACKGROUND: Leptospirosis is among the leading zoonotic causes of morbidity and mortality worldwide. Knowledge about spatial patterns of diseases and their underlying processes have the potential to guide intervention efforts. However, leptospirosis is often an underreported and misdiagnosed disease and consequently, spatial patterns of the disease remain unclear. In the absence of accurate epidemiological data in the urban agglomeration of Santa Fe, we used a knowledge-based index and cluster analysis to identify spatial patterns of environmental and socioeconomic suitability for the disease and potential underlying processes that shape them. METHODS: We geocoded human leptospirosis cases derived from the Argentinian surveillance system during the period 2010 to 2019. Environmental and socioeconomic databases were obtained from satellite images and publicly available platforms on the web. Two sets of human leptospirosis determinants were considered according to the level of their support by the literature and expert knowledge. We used the Zonation algorithm to build a knowledge-based index and a clustering approach to identify distinct potential sets of determinants. Spatial similarity and correlations between index, clusters, and incidence rates were evaluated. RESULTS: We were able to geocode 56.36% of the human leptospirosis cases reported in the national epidemiological database. The knowledge-based index showed the suitability for human leptospirosis in the UA Santa Fe increased from downtown areas of the largest cities towards peri-urban and suburban areas. Cluster analysis revealed downtown areas were characterized by higher levels of socioeconomic conditions. Peri-urban and suburban areas encompassed two clusters which differed in terms of environmental determinants. The highest incidence rates overlapped areas with the highest suitability scores, the strength of association was low though (CSc r = 0.21, P < 0.001 and ESc r = 0.19, P < 0.001). CONCLUSIONS: We present a method to analyze the environmental and socioeconomic suitability for human leptospirosis based on literature and expert knowledge. The methodology can be thought as an evolutive and perfectible scheme as more studies are performed in the area and novel information regarding determinants of the disease become available. Our approach can be a valuable tool for decision-makers since it can serve as a baseline to plan intervention measures.

Seroprevalence of leptospiral antibodies in rodents from riverside communities of Santa Fe, Argentina.

BACKGROUND: Leptospirosis is a zoonotic disease that can be transmitted by contact with the urine of infected mammals. Rodents play a mayor role in the transmission of leptospires to humans. The province of Santa Fe reports the greatest number of cases in Argentina. Yet, in this region, there are still knowledge gaps regarding the diversity of rodent species that may be hosts of pathogenic leptospires. The aims of this study were to evaluate the presence of leptospiral antibodies in rodents from three riverside communities of Santa Fe, and to identify factors associated with leptospiral infection. METHODOLOGY/PRINCIPAL FINDINGS: Each community was divided into three environmental settings based on the level of human disturbance, and sampled during two springs (Sep-Oct 2014 and 2015) and one autumn (Mar-Apr 2015). Serum samples of captured sigmodontine and murine rodents were tested for leptospiral antibodies by enzyme-linked immunosorbent assay (ELISA), and microagglutination test (MAT) was used to assess the infecting serovar in seropositive individuals. Factors influencing seropositivity were analyzed using logistic regression models. We caught 119 rodents, of which 101 serums were suitable for analysis. Most frequently trapped species were Scapteromys aquaticus, Akodon azarae and Oligoryzomys spp., with seroprevalences of 41.3%, 42.9% and 55% respectively. Seropositivity was higher in individuals with an average body condition score and in those that were sexually mature, but in the latter the differences were marginally significant. CONCLUSIONS/SIGNIFICANCE: Our results suggest that native rodents may be playing a role in the environmental circulation of pathogenic leptospires and provide relevant information for public health policies in the area.

Leptospirosis humana en Argentina: un esquema de análisis, 2014 / Human Leptospirosis in Argentina: An Analysis Scheme, 2014

INTRODUCCIÓN: En general, sólo existen estimaciones sobre el número de casos de leptospirosis en las Américas. La fuente más común son los Ministerios de Salud, tanto nacionales como provinciales, que proporcionan datos útiles sobre tendencias de incidencia de la leptospirosis, identificación de brotes y efectos de intervenciones gubernamentales. OBJETIVOS: Conocer la incidencia de casos de leptospirosis detectados por laboratorio durante 2014 en Argentina y generar un esquema de análisis de bases de datos de diferentes organismos nacionales para que sea repetido y difundido anualmente. MÉTODOS: Se analizaron las bases de datos del Sistema de Vigilancia Laboratorial (SIVILA)y del Instituto Nacional de Enfermedades Respiratorias (INER), teniendo en cuenta los casos notificados en 2014, según la fecha de inicio de síntomas. RESULTADOS: La incidencia de casos confirmados de leptospirosis en 2014 fue de 0,32/100 000 habitantes, mayormente detectados en las provincias de Santa Fe, Buenos Aires y Entre Ríos. Los serogrupos circulantes más prevalentes fueron Icterohaemorrhagiae, Sejroe y Canicola. CONCLUSIONES: Se resalta la utilidad de este tipo de estudios, que incentivan la búsqueda de casos, notificación y vigilancia de la leptospirosis, tanto para mejorar el conocimiento de la incidencia de la enfermedad y su distribución como para orientar las acciones hacia los lugares de mayor riesgo del país.

INTRODUCTION: In general, there are only estimates of the number of cases in the Americas. This data is commonly obtained by both national and provincial Ministries of Health which, despite data limitations, provide useful information on leptospirosis incidence trends, outbreak identification and effects of government interventions. OBJECTIVES: To know the incidence of leptospirosis cases detected by laboratory during 2014 in Argentina, and to generate an analysis scheme of databases from different national organisms to be repeated and diffused annually. METHODS: The SIVILA and INER databases were analyzed, taking into account the cases reported in 2014, according to the date of onset of symptoms. RESULTS: The incidence of confirmed leptospirosis cases in 2014 was 0.32/100 000 persons, mostly detected in Santa Fe, Buenos Aires and Entre Ríos provinces. The most prevalent circulating serogroups were Icterohaemorrhagiae, Sejroe and Canicola. CONCLUSIONS: These studies motivate the suspicion, notification and surveillance of leptospirosis and are useful both to improve the knowledge of the incidence of cases and their distribution, as well as to guide actions towards the most risky places in the country.

Clinical Evaluation of Rapid Diagnostic Test Kit Using the Polysaccharide as a Genus-Specific Diagnostic Antigen for Leptospirosis in Korea, Bulgaria, and Argentina.

Leptospirosis, a zoonotic disease that is caused by many serovars which are more than 200 in the world, is an emerging worldwide disease. Accurate and rapid diagnostic tests for leptospirosis are a critical step to diagnose the disease. There are some commercial kits available for diagnosis of leptospirosis, but the obscurity of a species- or genus-specific antigen of pathogenic Leptospira interrogans causes the reduced sensitivity and specificity. In this study, the polysaccharide derived from lipopolysaccharide (LPS) of nonpathogenic Leptospira biflexa serovar patoc was prepared, and the antigenicity was confirmed by immunoblot and enzyme linked immunosorbent assay (ELISA). The performance of the rapid diagnostic test (RDT) kit using the polysaccharide as a diagnostic antigen was evaluated in Korea, Bulgaria and Argentina. The sensitivity was 93.9%, 100%, and 81.0% and the specificity was 97.9%, 100%, and 95.4% in Korea (which is a rare region occurring with 2 serovars mostly), Bulgaria (epidemic region with 3 serovars chiefly) and Argentina (endemic region with 19 serovars mainly) respectively. These results indicate that this RDT is applicable for global diagnosis of leptospirosis. This rapid and effective diagnosis will be helpful for diagnosis and manage of leptospirosis to use and the polysaccharide of Leptospira may be called as genus specific antigen for diagnosis.

Isolation and clinical sample typing of human leptospirosis cases in Argentina.

Leptospira typing is carried out using isolated strains. Because of difficulties in obtaining them, direct identification of infective Leptospira in clinical samples is a high priority. Multilocus sequence typing (MLST) proved highly discriminatory for seven pathogenic species of Leptospira, allowing isolate characterization and robust assignment to species, in addition to phylogenetic evidence for the relatedness between species. In this study we characterized Leptospira strains circulating in Argentina, using typing methods applied to human clinical samples and isolates. Phylogenetic studies based on 16S ribosomal RNA gene sequences enabled typing of 8 isolates (6 Leptospira interrogans, one Leptospira wolffii and one Leptospira broomii) and 58 out of 85 (68.2%) clinical samples (55 L. interrogans, 2 Leptospira meyeri, and one Leptospira kirschneri). MLST results for the L. interrogans isolates indicated that five were probably Canicola serogroup (ST37) and one was probably Icterohaemorrhagiae serogroup (ST17). Eleven clinical samples (21.6%), provided MLST interpretable data: five were probably Pyrogenes serogroup (ST13), four Sejroe (ST20), one Autumnalis (ST22) and one Canicola (ST37). To the best of our knowledge this study is the first report of the use of an MLST typing scheme with seven loci to identify Leptospira directly from clinical samples in Argentina. The use of clinical samples presents the advantage of the possibility of knowing the infecting strain without resorting to isolates. This study also allowed, for the first time, the characterization of isolates of intermediate pathogenicity species (L. wolffii and L. broomii) from symptomatic patients.

[Evaluation of a reduced panel of leptospira strains for microagglutination]. / Evaluación de un panel reducido de cepas de leptospiras para el diagnóstico en humanos por microaglutinación (MAT).

OBJECTIVE: To evaluate if the use of the 19 Leptospira strains panel suggested by the International Leptospirosis Society of World Health Organization for microagglutination allows confirmation of more cases that the 12 strains panel used in Argentina. MATERIALS AND METHODS: Cross-sectional observational study. We studied 441 serum samples corresponding to Argentinean patients with suspected leptospirosis derived during from July to December, 2009 and from January to October, 2013. RESULTS: The same number of positive samples was obtained using the MAT with the 19 or 12 strains. In six cases a serovar of the expanded collection was presumably infecting, but always coagglutinated with strains of the reduced panel. CONCLUSION: In Argentina, the diagnosis of leptospirosis by MAT could be made using the reduced 12 strains panel, obtaining the same result in case detection as using the 19 strains panel. Additional information provided by the use of all strains could be the presumably infecting serogroup.

Evaluación de un panel reducido de cepas de leptospiras para el diagnóstico en humanos por microaglutinación (MAT) / Evaluation of a reduced panel of leptospira strains for microagglutination

Objetivo. Evaluar si el uso del panel de 19 cepas de leptospiras, sugerido por la Sociedad Internacional de Leptospirosis para la microaglutinación (MAT, por sus siglas en inglés), permite mayor confirmación de casos que el de 12 cepas. Material y métodos. Estudio observacional de corte transversal. Se estudiaron 441 muestras de sueros de pacientes de Argentina, derivadas para el diagnóstico de leptospirosis en los periodos de julio de 2009 a diciembre de 2010 y enero a octubre de 2013. Resultados. Se obtuvo el mismo resultado con el panel reducido que con el ampliado. En seis casos resultó presumiblemente infectante algún serovar del panel ampliado, aunque siempre coaglutinando con cepas del reducido. Conclusión. En Argentina, el diagnóstico de leptospirosis por MAT podría continuar realizándose con el panel reducido, lo que reduciría el costo y tiempo de diagnóstico. La información adicional que aportaría el panel ampliado está relacionada con la epidemiología, mediante un mejor conocimiento del serogrupo presumiblemente infectante.

Objective. To evaluate if the use of the 19 Leptospira strains panel suggested by the International Leptospirosis Society of World Health Organization for microagglutination allows confirmation of more cases that the 12 strains panel used in Argentina. Materials and methods. Cross-sectional observational study. We studied 441 serum samples corresponding to Argentinean patients with suspected leptospirosis derived during from July to December, 2009 and from January to October, 2013. Results. The same number of positive samples was obtained using the MAT with the 19 or 12 strains. In six cases a serovar of the expanded collection was presumably infecting, but always coagglutinated with strains of the reduced panel. Conclusion. In Argentina, the diagnosis of leptospirosis by MAT could be made using the reduced 12 strains panel, obtaining the same result in case detection as using the 19 strains panel. Additional information provided by the use of all strains could be the presumably infecting serogroup.

Aspectos ecológicos y operacionales de la vigilancia de leptospirosis em Argentina / Ecological and operational aspects of the surveillance of leptospirosis in Argentina

La leptospirosis es una de las zoonosis más ampliamente distribuidas en elmundo, y continúa siendo un problema de salud pública, especialmente en países donde ocurren abundantes precipitaciones. En Argentina, si bien el trabajo rural constituye un factor de riesgo importante, ha emergido como principal factor de riesgo el contacto con inundaciones por lluvias, especialmente en grandes centros urbanos. El objetivo principal del trabajo fue identificar patrones espacio-temporales de la transmisión de leptospirosis en Argentina, en base a datos de los casos de leptospirosis probables y confirmados desde el año 2009 a la actualidad, y su relación con las condiciones geográficas, ambientales y operacionales del sistema de laboratorios y vigilancia epidemiológica. Para hacerlo, se crearon nuevas regiones de estudio utilizando un software GIS y se construyeron las series de tiempo de casos para cada una. Se analizaron según sus componentes y se realizó una estimación del modelo de factores, resultando en 1 factor explicativo. A través de ello se detectaron tres padrones diferentes de transmisión de leptospirosis en Argentina, mostrando que posiblemente estén definidos por la diferencia de climas. Además, el análisis de los datos de la vigilancia mostró que existe gran porcentaje de casos no resueltos, y zonas silenciosas que deberían notificar casos sin embargo no lo hacen...

Leptospirosis is the most widespread zoonotic disease in the world, and it remains being a public health problem, especially in countries with heavy rains occurrence. In Argentina, while the rural labor is an important risk factor, contact with flooding rainshas emerged as a major risk factor, especially in large urban centers. The main objective was to identify spatio temporal transmission patterns of leptospirosis in Argentina, based on data of probable and confirmed leptospirosis cases since 2009 to the present, and its relation to geographical, environmental and operational conditions of laboratories system and epidemiological surveillance. To do this, new study regions were create dusing a GIS software and were constructed the time series of cases for each. The series were analyzed according to their components and the estimated factor model was performed, resulting in one explanatory factor. Three different transmission standards of leptospirosis were detected in Argentina, showing that they are possibly defined by the different climates. Furthermore, analysis of surveillance data showed that there is large percentage of unsolved cases and silent zones that should report cases...

Diseño y aplicación de una nueva herramienta para la evaluación de la Red Nacional de Laboratorios de Leptospirosis / Design and application of a new tool for the evaluation of the national betwork of leptospirosis laboratories

INTRODUCCION: La leptospirosis es unaenfermedad febril aguda, frecuentemente subdiagnosticada ysubnotificada por su diversa presentación clínica, baja sospechay falta de disponibilidad de métodos diagnósticos rápidos, simples y eficientes. El Laboratorio Nacional de Referencia de Leptospirosis (LNRL) del Instituto Nacional de EnfermedadesRespiratorias (INER) ôDr. Emilio Coniö produce antígeno termorresistente (TR) y lo distribuye a los laboratorios de la Red Nacional de Laboratorios de Leptospirosis (RNLL). Luego, éstosrealizan la técnica y envían las muestras positivas al LNRL para la confirmación del diagnóstico y las negativas para control de calidad (CC). Sin embargo, hasta la realización de este estudio noexistía un control de uso de TR y envío de muestras. OBJETIVOS: Diseñar y aplicar una herramienta para evaluar operativamente la RNLL. METODOS: Durante 2012 y 2013 se aplicó una planilla de registro de uso de TR, y se crearon y analizaron indicadores de eficiencia de uso, aplicación y reposición del TR y derivación de muestras. RESULTADOS: Se observó que entre 2012 y 2013 mejoró la participación en el CC y la eficiencia de uso del TR.La derivación de muestras positivas y la solicitud de reposición de TR se mantuvieron similares. CONCLUSIONES: La planilladiseñada permitió obtener información de cantidad y calidad de uso del TR, y calcular indicadores para evaluar su distribución y aplicación. Se observó que el desempeño de los laboratoriosmejoró de un año al otro.

INTRODUCTION: Leptospirosis is an acute febrile disease, often underdiagnosed and underreported because of its diverse clinical presentation, low suspicion, and unavailability of rapid, simple and efficient diagnostic methods. TheNational Leptospirosis Reference Laboratory (LNRL) of the National Institute of Respiratory Diseases ôDr. Emilio Coniö produces thermo-resistant antigen (TR) and distributes it to the National Network of Leptospirosis Laboratories (RNLL). These laboratories perform the screening, send the positivesamples for confirmation and the negative ones for quality control. However, until this study there was no control of TR uses and shipment of samples. OBJECTIVES: To design andimplement a tool to operatively assess the RNLL. METHODS: During the years 2012 and 2013, a registration form for the TR use was applied, also creating and analyzing indicators of use efficiency, application and replacement of TR, and referral of samples. RESULTS: It was noted that in the period2012-2013, the participation in the CC and efficiency of use of TR improved. The derivation of positive samples and request for replacement TR remained similar. CONCLUSIONS: The designed form allowed for the first time to obtain information on the quantity and quality of use of the reagent, to design andcalculate indicators to assess the distribution and application of TR during 2012 and 2013. In general, the performance of decentralized laboratories improved from one year to the next.

Vigilancia intensificada de Leptospirosis en Santa Fe y Entre Ríos (2012-2013) / Leptospirosis Enhanced Surveillance System in Santa Fe and Entre Rios (2012-2013)

La leptospirosis es la zoonosis de mayor prevalencia mundial. Santa Fe y Entre Ríos concentran la mayoría de casos en Argentina. OBJETIVO: Implementar y describir un sistema de vigilancia intensificada de casos de leptospirosis en Santa Fe y Entre Ríos. MÉTODOS: La investigación, desarrollada desde enero de 2012 hasta marzo de 2013, implicó la intervención en sistemas y servicios de salud, así como el fortalecimiento de la red de laboratorios para el diagnóstico específico y aislamiento de leptospiras. La información obtenida a partir de las fichas clínico-epidemiológicas de los casos confirmados fue analizada y presentada mediante medidas de resumen. La vigilancia intensificada se implementó en siete hospitales estratégicos. RESULTADOS: Ingresaron 183 pacientes. Se confirmaron 24 casos (13%): 10 por MAT, 9 por PCR en tiempo real y 5 por ambos métodos. Se obtuvieron 3 aislamientos (sero grupo Canicola). Fallecieron 4 pacientes con hemorragia pulmonar sumada a compromiso renal, hepático y/o plaquetopenia. CONCLUSIONES: La vigilancia intensificada permitió obtener aislamientos humanos, ratificó el valor de la MAT, evidenció la utilidad de PCR en tiempo real para diagnóstico precoz y corroboró la dificultad de obtener segundas muestras. La presentación clínica evidenció una elevada mortalidad global y una alta frecuencia de compromiso respiratorio asociado a disfunciones orgánicas múltiples de aparición precoz...

Leptospirosis is the most prevalent zoonosis worldwide. In Argentina, Santa Fe and Entre Ríos are the provinces where most cases occur. OBJECTIVE:To implement and describe a system of enhanced surveillance of leptospirosis in Santa Fe and Entre Ríos. METHODS: The enhanced surveillance was carried out from January 2012 to March 2013. The study involved the intervention in health services and systems as well as the laboratory network strengthening for specific diagnosis and leptospira isolation. The information collected from clinical epidemiological records of confirmed cases was analyzed and presented through summary measures. The surveillance was implemented in seven strategic hospitals. RESULTS: A total of 183 patients were enrolled, and 24 cases (13%)were confirmed: 10 through MAT, 9 through real time PCR and 5 through both methods. It was possible to obtain 3 isolates (Canicola serogroup). 4 patients died with pulmonary hemorrhage coupled with renal impairment, hepaticand/ or thrombocytopenia. CONCLUSIONS: The enhanced surveillance allowed to obtain human isolates, confirmed the diagnostic value of MAT, showed the utility of real time PCR for early diagnosis and the difficulties to obtain second samples. The clinical presentation revealed a high global mortality and a high frequency of respiratory involvement related to multiple organ dysfunctions from early stages...