An rK28-Based Immunoenzymatic Assay for the Diagnosis of Canine Visceral Leishmaniasis in Latin America.

Direct observation of Leishmania parasites in tissue aspirates has shown low sensitivity for the detection of canine visceral leishmaniasis (VL). Therefore in the last quarter century immunoenzymatic tests have been developed to improve diagnosis. The purpose of this study was to develop a fast recombinant K28 antigen, naked-eye qualitative enzyme-linked immunosorbent assay (VL Ql-ELISA) and a quantitative version (VL Qt-ELISA), and to display it in a kit format, whose cutoff value (0.156) was selected as the most adequate one to differentiate reactive from nonreactive samples. Considering 167 cases and 300 controls, sensitivity was 91% for both assays and specificity was 100% and 98.7% in Ql-ELISA and Qt-ELISA, respectively. Positive predictive values were 100% and 97.4% for Ql-ELISA and Qt-ELISA, respectively, and negative predictive values were 95.2% for both ELISAs. Reagent stability, reliability studies, including periodic repetitions and retest of samples, cutoff selection, and comparison of rK28 ELISAs with rK39 immunochromatographic test, were the international criteria that supported the quality in both kits. The performance of both ELISA kits in this work confirmed their validity and emphasized their usefulness for low-to-medium complexity laboratories.

Leishmaniasis visceral: senderos que confluyen, se bifurcan / Visceral leishmaniasis: paths that converge and divide

La leishmaniasis visceral urbana es una zoonosis emergente en Argentina. En América es producida por Leishmania infantum, con el perro como reservorio principal e insectos flebotomíneos como vectores. En este artículo se presenta el conocimiento acumulado a partir de su emergencia y dispersión en el país, por los referentes del Programa Nacional de Leishmaniasis, en el diagnóstico clínico y de laboratorio, tratamiento, biología de vectores, manejo de reservorio, y el conflicto generado con las acciones recomendadas en relación con los perros infectados. La detección temprana y el tratamiento precoz, con estrategias descentralizadas y horizontales, contribuirán a disminuir la morbimortalidad asociada a la leishmaniasis visceral. El control de su transmisión y dispersión requiere de un manejo ambiental integral y la tenencia responsable de perros. Se discuten los intereses y discursos en conflicto generados por la leishmaniasis visceral en el marco de la relación humano-perro, proponiendo la búsqueda de un discurso consensuado de riesgo.(AU)

Urban visceral leishmaniasis is an emerging zoonosis in Argentina. In the Americas the disease is produced by Leishmania infantum, with dogs as the primary reservoir and phlebotomine sandflies as the vectors. This article presents the experience acquired by professionals from the National Leishmaniasis Program in Argentina as visceral leishmaniasis has emerged and spread, especially in clinical and laboratory diagnosis, disease treatment, vector biology, reservoir management, and conflicts regarding recommendations for infected dogs. Early detection and treatment, along with decentralized and horizontal strategies, will contribute to the decrease in morbidity and mortality associated with visceral leishmaniasis. Control over the transmission and spread of the disease requires integral environmental management and responsible dog ownership. The interests and discourses put into conflict by visceral leishmaniasis are discussed in the framework of the human-dog relationship, and the search for a consensus-based risk discourse is proposed.(AU)

Leishmaniasis visceral: senderos que confluyen, se bifurcan / Visceral leishmaniasis: paths that converge and divide

La leishmaniasis visceral urbana es una zoonosis emergente en Argentina. En América es producida por Leishmania infantum, con el perro como reservorio principal e insectos flebotomíneos como vectores. En este artículo se presenta el conocimiento acumulado a partir de su emergencia y dispersión en el país, por los referentes del Programa Nacional de Leishmaniasis, en el diagnóstico clínico y de laboratorio, tratamiento, biología de vectores, manejo de reservorio, y el conflicto generado con las acciones recomendadas en relación con los perros infectados. La detección temprana y el tratamiento precoz, con estrategias descentralizadas y horizontales, contribuirán a disminuir la morbimortalidad asociada a la leishmaniasis visceral. El control de su transmisión y dispersión requiere de un manejo ambiental integral y la tenencia responsable de perros. Se discuten los intereses y discursos en conflicto generados por la leishmaniasis visceral en el marco de la relación humano-perro, proponiendo la búsqueda de un discurso consensuado de riesgo.

Urban visceral leishmaniasis is an emerging zoonosis in Argentina. In the Americas the disease is produced by Leishmania infantum, with dogs as the primary reservoir and phlebotomine sandflies as the vectors. This article presents the experience acquired by professionals from the National Leishmaniasis Program in Argentina as visceral leishmaniasis has emerged and spread, especially in clinical and laboratory diagnosis, disease treatment, vector biology, reservoir management, and conflicts regarding recommendations for infected dogs. Early detection and treatment, along with decentralized and horizontal strategies, will contribute to the decrease in morbidity and mortality associated with visceral leishmaniasis. Control over the transmission and spread of the disease requires integral environmental management and responsible dog ownership. The interests and discourses put into conflict by visceral leishmaniasis are discussed in the framework of the human-dog relationship, and the search for a consensus-based risk discourse is proposed.

Leishmaniasis visceral: contribución a su control mediante el fortalecimiento del diagnóstico del reservorio canino en Posadas, Misiones, Argentina / Visceral leishmaniasis: control contribution by strengthening the diagnosis of dog reservoir in the city of Posadas, Misiones, Argentina

INTRODUCCION: La leishmaniasis visceral (LV) es una enfermedad emergente en Argentina. En 2006 se describió el primer caso autóctono en Posadas (Misiones), ciudad que se convirtió en epicentro de la LV.OBJETIVO: Determinar la prevalencia de la LV canica (LVC) y fortalecer su diagnóstico en Posadas.METODOS: Con un criterio entomológico, la ciudad de Posadas se dividió en islas de alta densidad (IAD) e islas de baja densidad (IBD) de flebótomos. Se seleccionó, de forma arbitraria y mediante imágenes satelitales, la manzana situada más al norte y más al sur de cada IAD e IBD. Se realizó un diseño observacional prospectivo, y se estudió a todos los perros de las manzanas seleccionadas. Se consideró un N inicial de 400 animales para detectar una diferencia de al menos 7% con una potencia de 0,90 y un nivel de 0,05 respecto de la prevalencia global supuesta del 20%. Se tomaron muestras de sangre, punción de ganglios linfáticos y raspado de piel. El estudio incluyó además un test serológico rápido (rK39 canino) y frotis de las muestras para búsqueda parasitológica, que se procesaron y evaluaron en el Instituto Municipal de Salud Animal de Posadas y en el Instituto Nacional de Parasitología Fatala Chabén.RESULTADOS: De los 182 animales evaluados, 33 fueron positivos para LVC, con una prevalencia global del 18,1%. En IAD la prevalencia fue 22% y en IBD, 13,4%. Los estudios parasitológicos fueron positivos en el 10% de los caninos en IAD y 1,2% en IBD. La LVC fue asintomática u oligosintomática, con una presentación más frecuente en IAD. La rK39 tuvo una sensibilidad del 90,3% y una especificidad del 98,9%.CONCLUSIONES: El análisis serológico con antígeno rK39 fue siempre más sensible que la búsqueda parasitológica para el diagnóstico. La LVC parece tener menor prevalencia en relación con estudios previos, quizás debido a diferentes medidas de control realizadas en el Municipio de Posadas.

INTRODUCCION: Visceral leishmanisis (VL) is an emerging disease in Argentina. The first autochthonous case was described in 2006 in a chld from Posadas (Misiones), the city which turned out to be the focus of VL.OBJECTIVE: To determine the prevalence of canine VL (CVL) and improve its diagnosis in Posadas.METHODS: According to entomological criteria, Posadas was divided in high density islands (HDI) and low density islands (LDI) of sandfly. Randomly and with satellite images, the northernmost and southernmost blocks of each HDI and LDI were selected. A prospective, experimental design was performed, studying all the dogs in the selected blocks. There was an initial sample of 400 animals to detect a difference of at least 7%, with a power of 0.90 and a level of 0.05, compared to the supposed prevalence of 20%. Blood samples were taken, with puncture of lymph nodes and skin scraping. The study also included a (canine rK39) rapid test for serological assessment adn sample smears for parasitological search. Samples were processed and evaluated in the Posadas institute of animal health (I.MU.S.A) and the Fatala Chabén National Institute of Parasitology.RESULTS: CVL was assessed in 33/182 animals with an overall prevalence of 18.1%. In HDI, the prevalence was 22%; in LDI, it was 13.4%. Parasitological studies were positive in 10% (HDI) and 1.2% (LDI). The CVL had a nonspecific clinical presentation; canines were asimptomatic or oligosymptomatic, with a higher trend in HDI. The rK39 had a sensitivity of 90.3% and a specificity of 98.9%.CONCLUSIONS: The antigen rK39 was always more sensitive than the search for parasites for the diagnosis of CVL. The disease appears to be less prevalent comapring to previous studies, perhaps due to different control measures performed in the Municipality of Posadas.

Leishmaniasis visceral: contribución a su control mediante el fortalecimiento del diagnóstico del reservorio canino en Posadas, Misiones, Argentina / Visceral leishmaniasis: control contribution by strengthening the diagnosis of dog reservoir in the city of Posadas, Misiones, Argentina

INTRODUCCION: La leishmaniasis visceral (LV) es una enfermedad emergente en Argentina. En 2006 se describió el primer caso autóctono en Posadas (Misiones), ciudad que se convirtió en epicentro de la LV.OBJETIVO: Determinar la prevalencia de la LV canica (LVC) y fortalecer su diagnóstico en Posadas.METODOS: Con un criterio entomológico, la ciudad de Posadas se dividió en islas de alta densidad (IAD) e islas de baja densidad (IBD) de flebótomos. Se seleccionó, de forma arbitraria y mediante imágenes satelitales, la manzana situada más al norte y más al sur de cada IAD e IBD. Se realizó un diseño observacional prospectivo, y se estudió a todos los perros de las manzanas seleccionadas. Se consideró un N inicial de 400 animales para detectar una diferencia de al menos 7% con una potencia de 0,90 y un nivel de 0,05 respecto de la prevalencia global supuesta del 20%. Se tomaron muestras de sangre, punción de ganglios linfáticos y raspado de piel. El estudio incluyó además un test serológico rápido (rK39 canino) y frotis de las muestras para búsqueda parasitológica, que se procesaron y evaluaron en el Instituto Municipal de Salud Animal de Posadas y en el Instituto Nacional de Parasitología Fatala Chabén.RESULTADOS: De los 182 animales evaluados, 33 fueron positivos para LVC, con una prevalencia global del 18,1%. En IAD la prevalencia fue 22% y en IBD, 13,4%. Los estudios parasitológicos fueron positivos en el 10% de los caninos en IAD y 1,2% en IBD. La LVC fue asintomática u oligosintomática, con una presentación más frecuente en IAD. La rK39 tuvo una sensibilidad del 90,3% y una especificidad del 98,9%.CONCLUSIONES: El análisis serológico con antígeno rK39 fue siempre más sensible que la búsqueda parasitológica para el diagnóstico. La LVC parece tener menor prevalencia en relación con estudios previos, quizás debido a diferentes medidas de control realizadas en el Municipio de Posadas.

INTRODUCCION: Visceral leishmanisis (VL) is an emerging disease in Argentina. The first autochthonous case was described in 2006 in a chld from Posadas (Misiones), the city which turned out to be the focus of VL.OBJECTIVE: To determine the prevalence of canine VL (CVL) and improve its diagnosis in Posadas.METHODS: According to entomological criteria, Posadas was divided in high density islands (HDI) and low density islands (LDI) of sandfly. Randomly and with satellite images, the northernmost and southernmost blocks of each HDI and LDI were selected. A prospective, experimental design was performed, studying all the dogs in the selected blocks. There was an initial sample of 400 animals to detect a difference of at least 7%, with a power of 0.90 and a level of 0.05, compared to the supposed prevalence of 20%. Blood samples were taken, with puncture of lymph nodes and skin scraping. The study also included a (canine rK39) rapid test for serological assessment adn sample smears for parasitological search. Samples were processed and evaluated in the Posadas institute of animal health (I.MU.S.A) and the Fatala Chabén National Institute of Parasitology.RESULTS: CVL was assessed in 33/182 animals with an overall prevalence of 18.1%. In HDI, the prevalence was 22%; in LDI, it was 13.4%. Parasitological studies were positive in 10% (HDI) and 1.2% (LDI). The CVL had a nonspecific clinical presentation; canines were asimptomatic or oligosymptomatic, with a higher trend in HDI. The rK39 had a sensitivity of 90.3% and a specificity of 98.9%.CONCLUSIONS: The antigen rK39 was always more sensitive than the search for parasites for the diagnosis of CVL. The disease appears to be less prevalent comapring to previous studies, perhaps due to different control measures performed in the Municipality of Posadas.

The activity of azithromycin against Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis in the golden hamster model

New therapeutic alternatives against leishmaniasis remain a priority. The activity of azithromycin against Leishmania (Leishmania) major has been previously demonstrated. Different responses among species of Leishmania make species-specific drug screening necessary. The activity of azithromycin against Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis was evaluated in golden hamsters infected through footpad injections of metacyclic promastigotes, and compared with untreated controls and animals treated with meglumine antimoniate. Footpad thickness, lesion cultures and dissemination sites were analyzed. Treatment of golden hamsters with oral azithromycin at 450mg/kg had no activity against infections with Leishmania (Leishmania) amazonensis. For infections due to Leishmania (Viannia) braziliensis, azithromycin demonstrated significant activity relative to untreated controls, but inferior to meglumine antimoniate, for controlling lesion size. Neither drug was able to totally eliminate parasites from the lesions. It was concluded that azithromycin has activity against Leishmania (Viannia) braziliensis but not against Leishmania (Leishmania) amazonensis in this model.

Novas alternativas terapêuticas contra a leishmaniose são ainda uma prioridade. A atividade da azitromicina contra a Leishmania (Leishmania) major foi anteriormente demonstrada. Diferentes respostas entre as espécies de Leishmania fazem com que um screening de drogas específicas para espécies seja necessário. A atividade da azitromicina contra a Leishmania (Viannia) braziliensis e a Leishmania (Leishmania) amazonensis foi avaliada em Golden hamsters infectados a través de injeções de promastigotas metacíclicas e comparando com controles sem tratamento e animais tratados com antimoniato de N-metil-glucamina. Foram analisadas a espessura da pata, a cultura das lesões e disseminação para órgãos internos. A azitromicina oral em dose de 450mg/kg não teve atividade contra a infecção por Leishmania ( Leishmania) amazonensis. Para infecções devidas à Leishmania (Viannia) braziliensis, a azitromicina teve uma atividade significativa em relação aos controles sem tratamento, mas foi inferior ao antimoniato de N-metil-glucamina quanto ao controle do tamanho das lesões. Nenhuma das drogas conseguiu eliminar totalmente os parasitos das lesões. Foi concluído que a azitromicina tem atividade contra Leishmania (Viannia) braziliensis, mas não tem atividade contra Leishmania (Leishmania) amazonensis neste modelo.

The activity of azithromycin against Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis in the golden hamster model.

New therapeutic alternatives against leishmaniasis remain a priority. The activity of azithromycin against Leishmania (Leishmania) major has been previously demonstrated. Different responses among species of Leishmania make species-specific drug screening necessary. The activity of azithromycin against Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis was evaluated in golden hamsters infected through footpad injections of metacyclic promastigotes, and compared with untreated controls and animals treated with meglumine antimoniate. Footpad thickness, lesion cultures and dissemination sites were analyzed. Treatment of golden hamsters with oral azithromycin at 450mg/kg had no activity against infections with Leishmania (Leishmania) amazonensis. For infections due to Leishmania (Viannia) braziliensis, azithromycin demonstrated significant activity relative to untreated controls, but inferior to meglumine antimoniate, for controlling lesion size. Neither drug was able to totally eliminate parasites from the lesions. It was concluded that azithromycin has activity against Leishmania (Viannia) braziliensis but not against Leishmania (Leishmania) amazonensis in this model.

Tegumentary leishmaniasis outbreak in Bella Vista City, Corrientes, Argentina during 2003.

Bella Vista City, Corrientes, Argentina, reported an epidemic outbreak of tegumentary leishmaniasis during 2003. The mean age of the 31 cases was 25.0 +/- 13.7 years old, with a sex ratio male:female 1.8, and without mucosal involvement. They clustered in two contiguous neighbourhoods, 96% in the periurban border and 4% in the peripheral outskirts. The transmission peak was estimated to have occurred during April 2003. Four species (3608 sand flies) were captured in nine sites: Lutzomyia neivai (90.1%), Lu. pessoai (8.9%), Lu. migonei (0.8 %), and Brumptomyia avellari (0.2 %). The outskirts/rural capture ratio of Lu. neivai was up to 3, and the outskirts/periurban up to 200. Therefore, the 'urban' transmission in this southernmost known focus is still an ecotone-border associated risk. The changes in human distribution or activities, patches of the secondary vegetation, periurban streams, rainfall of the previous year, and river period floods could all contribute to 'urban' outbreaks in the region. Tegumentary leishmaniasis risk should be assessed for any project that involves changes in land use throughout an endemic area.

Tegumentary leishmaniasis outbreak in Bella Vista City, Corrientes, Argentina during 2003

Bella Vista City, Corrientes, Argentina, reported an epidemic outbreak of tegumentary leishmaniasis during 2003. The mean age of the 31 cases was 25.0 ± 13.7 years old, with a sex ratio male:female 1.8, and without mucosal involvement. They clustered in two contiguous neighbourhoods, 96 percent in the periurban border and 4 percent in the peripheral outskirts. The transmission peak was estimated to have occurred during April 2003. Four species (3608 sand flies) were captured in nine sites: Lutzomyia neivai (90.1 percent), Lu. pessoai (8.9 percent), Lu. migonei (0.8 percent), and Brumptomyia avellari (0.2 percent). The outskirts/rural capture ratio of Lu. neivai was up to 3, and the outskirts/periurban up to 200. Therefore, the 'urban' transmission in this southernmost known focus is still an ecotone-border associated risk. The changes in human distribution or activities, patches of the secondary vegetation, periurban streams, rainfall of the previous year, and river period floods could all contribute to 'urban' outbreaks in the region. Tegumentary leishmaniasis risk should be assessed for any project that involves changes in land use throughout an endemic area.

Transmission of T. cruzi infection via liver transplantation to a nonreactive recipient for Chagas' disease.

Chagas' disease is an endemic zoonosis of South America caused by a protozoan parasite Trypanosoma cruzi. About 30% of infected people develop the disease. This disease is known to reactivate in immunocompromised hosts, such as patients with acquired immunodeficiency syndrome, leukemia, and transplantation. There is some experience with transplantation of infected renal grafts into negative recipients, resulting in an index of transmission of 35%. No cases have been reported involving other organ transplants up to 2002, when the Centers for Disease Control and Prevention reported 3 cases of Chagas' disease transmission to 3 recipients (liver, kidney, and pancreas-kidney) from a single chagas infected donor. Here we report on a case of orthotopic liver transplant from a chagas infected donor into a negative recipient in clinical emergency status. The recipient was monitored by direct parasitological Strout method and serological tests with detection of transmission on the 84 th day by both studies, without clinical signs. The patient was put on benznidazole with rapid clearance of the parasitemia. However, we propose that chagas infected donors may be accepted for liver transplant recipients only in emergency status.

Caracteristicas clinicas y diagnosticas de la leishmaniasis mucocutanea en pacientes de un area endemica de Salta / Clinical and diagnostic characteristics of mucocutaneous leishmaniasis in patients of an endemic area in Salta

Entre junio de 1990 y diciembre de 1992, en un área endémica para leishmaniasis, de Salta, Argentina, se examinaron 39 pacientes con diagnóstico clínico de leishmaniasis tegumentaria. El 87,2 por ciento (34/39) de los casos presentaron la forma cutánea simple, el 10,3 por ciento la forma cutánea múltiple y en 2.6 por ciento mucosa. La localización corporal más frecuente de las lesiones fue en miembros (71.8 por ciento), seguida de tronco y de la localización múltiple (10.3 por ciento). El 43.6 por ciento eran amas de casa, estudiantes o niños, sugiriendo que la infección pudo ser contraída en el ambiente doméstico o peridoméstico. De los 39 pacientes diagnosticados, en 22 (56.4 por ciento) se realizó la comprobación parasitológica. El examen del frotis, por microscopía directa, permitió el diagnóstico de 13 (59.4 por ciento) de los 22 pacientes. En 5 (22.7 por ciento se encontró leishmanias por cultivos, y por inoculación del hámster en 9 (40.9 por ciento). Se obtuvieron 10 aislamientos parasitarios (45.4 por ciento). Se recomienda, como método de diagnóstico para la vigilancia epidemiológica, la observación micorscópica directa del frotis, dada la sensibilidad demonstrada en este trabajo y la facilidad de su implementación en la endémica. El período de evolución clínica, desde la aparición de la lesión hasta la detección del paciente por el Sistema de Salud, fue de aproximadamente 90 días. Ese período estaría relacionado a la frecuencia de vista del Agente Sanitario a los domicilios, cada 3 meses. Sólo un paciente tratado tuvo una recidiva a los 6 meses, debido al incumplimiento del tratamiento. (AU)

Caracteristicas clinicas y diagnosticas de la leishmaniasis mucocutanea en pacientes de un area endemica de Salta / Clinical and diagnostic characteristics of mucocutaneous leishmaniasis in patients of an endemic area in Salta

Entre junio de 1990 y diciembre de 1992, en un área endémica para leishmaniasis, de Salta, Argentina, se examinaron 39 pacientes con diagnóstico clínico de leishmaniasis tegumentaria. El 87,2 por ciento (34/39) de los casos presentaron la forma cutánea simple, el 10,3 por ciento la forma cutánea múltiple y en 2.6 por ciento mucosa. La localización corporal más frecuente de las lesiones fue en miembros (71.8 por ciento), seguida de tronco y de la localización múltiple (10.3 por ciento). El 43.6 por ciento eran amas de casa, estudiantes o niños, sugiriendo que la infección pudo ser contraída en el ambiente doméstico o peridoméstico. De los 39 pacientes diagnosticados, en 22 (56.4 por ciento) se realizó la comprobación parasitológica. El examen del frotis, por microscopía directa, permitió el diagnóstico de 13 (59.4 por ciento) de los 22 pacientes. En 5 (22.7 por ciento se encontró leishmanias por cultivos, y por inoculación del hámster en 9 (40.9 por ciento). Se obtuvieron 10 aislamientos parasitarios (45.4 por ciento). Se recomienda, como método de diagnóstico para la vigilancia epidemiológica, la observación micorscópica directa del frotis, dada la sensibilidad demonstrada en este trabajo y la facilidad de su implementación en la endémica. El período de evolución clínica, desde la aparición de la lesión hasta la detección del paciente por el Sistema de Salud, fue de aproximadamente 90 días. Ese período estaría relacionado a la frecuencia de vista del Agente Sanitario a los domicilios, cada 3 meses. Sólo un paciente tratado tuvo una recidiva a los 6 meses, debido al incumplimiento del tratamiento.

Natural Trypanosoma cruzi infection in dogs of endemic areas of the Argentine Republic

Estudou-se a dinâmica populacional e a prevalência de infecçäo chagásica de 352 cäes vivendo em 108 moradias rurais infectadas por triatomíneos. A área foi dividida em três setores de acordo com suas crescentes distâncias em relaçäo a um povoado urbano. Cada animal foi identificado mediante suas características particulares, e seus donos deram informaçöes a respeito de seus hábitos. As prevalências de infecçäo foram determinadas por sorologia, correlaçäo parasitológica-sorológica, percentual de traçados eletrocardiográficos alterados e o percentual de moradias com cäes parasitados. A área rural mostrou um padräo característico de infecçäo por T. Cruzi e observaram-se diferenças nos parâmetros de populaçäo canina em relaçäo às restantes áreas: maior proporçäo de filhotes, maior populaçäo humana e canina sedentárias, maiores prevalências de infecçäo por xenodiagnóstico em cäes e maior proporçäo de barbeiros ingfectados com T. Cruzi nos cômodos da casa. Supöe-se que as características sedentárias da populaçäo humana desta área rural é fundamental para a oferta de sangue para as populaçöes triatomíneas, e a elevada porcentagem de cäes parasitêmicos dessa área contribui para o aumento de barbeiros infectados com T. Cruzi encontrados nos quartos de dormir