Seroprevalence of anti-Leptospira antibodies in equines and associated workers-Isolation of Leptospira interrogans serogroup Canicola from equine urine.

To investigate seroprevalence of anti-Leptospira antibodies in equines and associated workers in Uruguay, 891 equine and 150 human sera were drawn; 212 equine urine samples were also taken for culture. Environmental conditions and equine raising or managing practices were recorded in all 72 visited establishments; epidemiological information was obtained from each worker. Microscopic agglutination technique (MAT) was performed with 10 Leptospira strains for equines and 18 for human sera, that were also studied with IgM indirect immunofluorescence (IgM-IIF). Equine titres ≥100 were considered positive, and human sera titres ≥200 suggested probable recent or past infection. Urines were cultured in Ellinghausen-McCullough-Johnson-Harris (EMJH) media; local identification of one obtained isolate with lipL32 PCR, Multiple Locus Variable number tandem repeat Analysis and partial rrs gene sequencing, were completed at Institut Pasteur, Paris. Estimated reactivity was 61.3% for equines, which was higher than the studied bovine national levels (21%) and mainly observed with Icterohaemorrhagiae serogroup (40.3%), Sejroe, Canicola, Pomona or Ballum. Aged animals from slaughterhouses and cattle farms were the most frequently positive. Multiple regression analysis confirmed a significant association between seropositivity and equine age. Only one positive culture could be fully studied, and confirmed to be Leptospira interrogans serogroup Canicola; it was added to the MAT antigen panel and revealed fairly frequent reaction with equine and human sera. Three workers (2%) showed titres = 200 with Icterohaemorrhagiae or Canicola serogroups, without recent clinical manifestations. Their attended equines reacted with the same serogroups, suggesting common source infections or infection transmitted by equines. Three other humans yielded titres = 100, and none of the 150 showed an IgM-IIF-positive result. Equines seem not to be an important origin of regional human leptospirosis, except perhaps during acute animal infection. More culture work is required to study intensity and lapses of leptospiruria, as well as to further identify circulating strains.

Seroprevalence of leptospirosis in human groups at risk due to environmental, labor or social conditions.

Leptospirosis is important in Uruguay due to the economic loss caused by the diseases of production animals, mainly bovines, and also due to frequent human infection. We decided to study anti-Leptospira antibodies in the sera of dairy workers, rice laborers, veterinarians, suburban slum dwellers and garbage recyclers. Our aims were to estimate the seroprevalence of infection by Leptospira spp. in these people at risk, the relative importance of the known risk factors associated with infection, and the impact of human infections in each setting. Groups at risk were identified and 35 visits to their locations were made, conducting field surveys and exchange talks for information and education. Simple epidemiological questionnaires were administered and sera samples were taken from 308 persons. The microagglutination Technique (MAT) and the IgM Indirect Immunofluorescence (IIF) assay were employed to detect antibodies. Environmental water samples, canine and equine sera were also examined. More than 45% of human sera were reactive and the studied groups were confirmed to be widely exposed to infection. Female sera were frequently reactive, though most illnesses occur in men, and the most severe cases in elderly males; the emergence and evolution of the disease may strongly depend on the host condition and functions. Animal contact and unsafe water usage were the main identified risk factors to be considered in prevention. Fifty per cent of the studied horses showed a positive MAT reaction. The underdiagnosis of the illness and its long-term symptoms require further study, as well as greater health and social attention efforts.

Isolation of pathogenic Leptospira strains from naturally infected cattle in Uruguay reveals high serovar diversity, and uncovers a relevant risk for human leptospirosis.

Leptospirosis is a neglected zoonosis with worldwide distribution. The causative agents are spirochete bacteria of the Leptospira genus, displaying huge diversity of serovars, the identity of which is critical for effective diagnosis and vaccination purposes. Among many other mammalian species, Leptospira infects cattle, eliciting acute signs in calves, and chronic disease in adult animals often leading to abortions. In South America, and including in Uruguay, beef and dairy export are leading sources of national income. Despite the importance of bovine health, food safety, and bovine-related dissemination of leptospirosis to humans, extremely limited information is available as to the identity of Leptospira species and serovars infecting cattle in Uruguay and the South American subcontinent. Here we report a multicentric 3-year study resulting in the isolation and detailed characterization of 40 strains of Leptospira spp. obtained from infected cattle. Combined serologic and molecular typing identified these isolates as L. interrogans serogroup Pomona serovar Kennewicki (20 strains), L. interrogans serogroup Canicola serovar Canicola (1 strain), L. borgpetersenii serogroup Sejroe serovar Hardjo (10 strains) and L. noguchii (9 strains). The latter showed remarkable phenotypic and genetic variability, belonging to 6 distinct serogroups, including 3 that did not react with a large panel of reference serogrouping antisera. Approximately 20% of cattle sampled in the field were found to be shedding pathogenic Leptospira in their urine, uncovering a threat for public health that is being largely neglected. The two L. interrogans serovars that we isolated from cattle displayed identical genetic signatures to those of human isolates that had previously been obtained from leptospirosis patients. This report of local Leptospira strains shall improve diagnostic tools and the understanding of leptospirosis epidemiology in South America. These strains could also be used as new components within bacterin vaccines to protect against the pathogenic Leptospira strains that are actually circulating, a direct measure to reduce the risk of human leptospirosis.

Characterization of Leptospira isolates from humans and the environment in Uruguay.

Laboratory diagnosis of human leptospirosis usually relies on indirect methods exploring specific immune response. Isolation and identification of the involved strains are cumbersome, but can provide biological resources for pathogenic studies and relevant information for guiding prevention and control measures. The aim of the research we are hereby reporting was the characterization of Leptospira isolates obtained from humans and the environment in Uruguay. Blood cultures were performed from early samples of 302 Uruguayan patients, mainly rural workers, and from 36 water samples taken from their living or working environments. Eight human isolates and seven environmental isolates were obtained and analyzed by end point Polymerase Chain Reaction (PCR), Multilocus Variable Number of Tandem Repeat Analysis (MLVA) and other molecular methods. Human isolates corresponded to several serogroups and serovars of Leptospira interrogans and Leptospira kirschneri species, probably reflecting the infection with similar involved Leptospira species and serovars of an extended animal reservoir in rural settings of the country, mostly dedicated to meat and dairy production. Culture-positive patients were older than usually affected workers, and presented signs and symptoms of severe illness. A high organic and circulating bacterial burden may explain an easier positive result from these workers' samples. Environmental isolates were mainly identified as Leptospira biflexa strains, with a single L. meyeri isolate of uncertain significance.

Valoración de un procedimiento de inmunofluorescencia indirecta para la detección de anticuerpos tipo IgM (IF-IgM) utilizado en el diagnóstico temprano de leptospirosis / Evaluation of an indirect immunofluorescence assay for detection of IgM (IF-IgM) type antibodies in the early diagnosis of leptospirosis

Introducción: la leptospirosis es una enfermedad febril, aguda, que presenta manifestaciones clínicas variadas. Esto dificulta o retarda el diagnóstico clínico, por lo cual es útil disponer de métodos de laboratorio adecuados para orientar el manejo inicial de estos pacientes. Objetivo: evaluar un procedimiento de inmunofluorescencia para detectar IgM (IF-IgM) de elaboración propia aplicado al diagnóstico temprano de leptospirosis. Material y método: se analizaron por IF-IgM y microaglutinación (MAT) (tomada como estándar de referencia) sueros obtenidos de pacientes con sospecha clínica de leptospirosis. La sensibilidad y especificidad de la IF-IgM versus MAT se establecieron utilizando una tabla de doble entrada. La concordancia entre dos observadores se determinó por el test Kappa. Resultados: de 161 muestras precoces analizadas, 97 sueros correspondieron a pacientes con infección aguda confirmada por MAT y 64 sin infección. La sensibilidad y especificidad de la IF-IgM con sueros de fase aguda fueron 79% y 100%, respectivamente. El índice Kappa fue 1. Conclusiones: la IF-IgM aparece como una herramienta útil para el diagnóstico temprano de pacientes con leptospirosis. No requiere el manejo de bacterias viables, puede realizarse en laboratorios que cuenten con microscopio de luz ultravioleta, se necesita una sola muestra de suero y el resultado está listo en tres a cuatro horas. En cuanto a las desventajas, no identifica los serovares involucrados y un resultado negativo no descarta la infección. Teniendo en cuenta esto último es obligatorio analizar por MAT una segunda muestra de suero obtenida a los 10-20 días de la primera para descartarla o confirmarla...