Chagas Disease and Leishmaniasis, Sympatric Zoonoses Present in Human from Rural Communities of Venezuela.

PURPOSE: Trypanosoma cruzi and Leishmania spp. coexist in several endemic areas, and there are few studies of Chagas disease and leishmaniasis coinfection worldwide; for this reason, the objective of this work was to determine the Chagas disease and leishmaniasis coinfection in several rural communities co-endemic for these diseases. METHODS: A total of 1107 human samples from six co-endemic rural communities of Cojedes state, Venezuela, were analyzed. Serum samples were evaluated by ELISA, indirect hemagglutination, and indirect immunofluorescence for Chagas disease diagnosis, and individuals were evaluated for leishmaniasis by leishmanin skin test (LST). Approximately, 30% of the individuals were also analyzed by PCR (blood clot samples) for T. cruzi and for Leishmania spp. RESULTS: The 14.7% of the individuals were positive to Trypanosoma cruzi infection by serology, and 25.8% were positive to Leishmania spp. current or past infection by LST. Among the group with PCR results, 7.8% were positive for T. cruzi, and 9.4% for Leishmania spp. The coinfection T. cruzi/Leishmania spp. was 6.5%. The T. cruzi DTUs of the positive blood clot samples were TcI, revealed using the molecular markers: (i) intergenic region of the miniexon, (ii) D7 divergent domain of the 24Sα rDNA, (iii) size-variable domain of the 18S rDNA, and (iv) hsp60-PCR-RFLP (EcoRV). The Leishmania species identified were L. (Leishmania) mexicana and L. (Viannia) braziliensis. CONCLUSION: A high prevalence was found for T. cruzi and Leishmania spp. single and coinfections in almost all communities studied, being these results of relevance for the implementation of control programs in co-endemic areas.

Surveillance for Leishmania asymptomatic infection in endemic foci of cutaneous leishmaniasis in Venezuela: a combination of leishmanin skin test and PCR using blood clots improves detection and enables identification of species.

BACKGROUND: Little is known about the prevalence of asymptomatic leishmaniasis in Venezuela. The objective of this study was to quantify Leishmania asymptomatic infection in six endemic foci of cutaneous leishmaniasis (CL) in Portuguesa State, Venezuela, where no previous data were available. METHODS: Study of the prevalence of Leishmania asymptomatic infection was carried out in 841 individuals from six endemic foci of CL in the municipalities Sucre and Ospino, Portuguesa State. We applied the leishmanin skin test (LST) and the internal transcribed spacer 1 (ITS1) PCR to DNA from sera and blood clots of all LST-positive and 20% of LST-negative patients. RESULTS: Of 841 inhabitants tested by LST, 197 returned a positive reaction (23.42%); all of the LST-positives (197) and 121 negatives were screened by nested PCR using serum and blood clots. Among the LST-positive group, 2.54% were PCR-positive with sera, while 44.67% were positive with blood clots. In the LST-negative group, PCR was positive in 2.48% of serum samples and in 38.84% of blood clots. CONCLUSIONS: It is recommended that LST and PCR on blood clots are used together to detect exposure and asymptomatic infection and for identification of the Leishmania species.

Molecular typing reveals the co-existence of two transmission cycles of American cutaneous leishmaniasis in the Andean Region of Venezuela with Lutzomyia migonei as the vector.

BACKGROUND The transmission routes for American cutaneous leishmaniasis (ACL) are in flux, so studies examining its transmission in humans, mammalian hosts, and sand fly vectors are urgently needed. OBJECTIVES The aim of this work was understand the epidemiological cycles of Leishmania spp., which causes ACL in the Andean Region of Venezuela, by identifying the Leishmania and the sand fly species involved in human and dog infections. METHODS Thirty-one biopsies from patients in Mérida and Táchira states with suspected ACL were studied by both parasitological tests (cultures and hamster inoculation) and a molecular test [Internal transcribed spacer 1 (ITS1) nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)]. We also conducted a survey to detect Leishmania infection in dogs (Immunifluorescence antibody test and ITS1 nested PCR-RFLP) and sand flies (ITS1 nested PCR-RFLP) from El Carrizal, a highly endemic focus of ACL in Venezuela. FINDINGS Three different Leishmania species were identified in the clinical samples from humans (Leishmania braziliensis, L. guyanensis, and L. mexicana) and dogs (L. guyanensis and L. mexicana). The predominant sand fly species found were those from the Verrucarum group (infected with L. mexicana) and Lutzomyia migonei (infected with L. guyanensis and L. mexicana). MAIN CONCLUSIONS We show that Lu. migonei may be the putative vector in two ACL epidemiological cycles, involving L. guyanensis and L. mexicana. We also report for the first time the presence of L. guyanensis in domestic animals.

Molecular typing reveals the co-existence of two transmission cycles of American cutaneous leishmaniasis in the Andean Region of Venezuela with Lutzomyia migonei as the vector

BACKGROUND The transmission routes for American cutaneous leishmaniasis (ACL) are in flux, so studies examining its transmission in humans, mammalian hosts, and sand fly vectors are urgently needed. OBJECTIVES The aim of this work was understand the epidemiological cycles of Leishmania spp., which causes ACL in the Andean Region of Venezuela, by identifying the Leishmania and the sand fly species involved in human and dog infections. METHODS Thirty-one biopsies from patients in Mérida and Táchira states with suspected ACL were studied by both parasitological tests (cultures and hamster inoculation) and a molecular test [Internal transcribed spacer 1 (ITS1) nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)]. We also conducted a survey to detect Leishmania infection in dogs (Immunifluorescence antibody test and ITS1 nested PCR-RFLP) and sand flies (ITS1 nested PCR-RFLP) from El Carrizal, a highly endemic focus of ACL in Venezuela. FINDINGS Three different Leishmania species were identified in the clinical samples from humans (Leishmania braziliensis, L. guyanensis, and L. mexicana) and dogs (L. guyanensis and L. mexicana). The predominant sand fly species found were those from the Verrucarum group (infected with L. mexicana) and Lutzomyia migonei (infected with L. guyanensis and L. mexicana). MAIN CONCLUSIONS We show that Lu. migonei may be the putative vector in two ACL epidemiological cycles, involving L. guyanensis and L. mexicana. We also report for the first time the presence of L. guyanensis in domestic animals.

Aspects of the bionomics of three phlebotomine vector species (Diptera: Psychodidae) at El Carrizal, an endemic focus of cutaneous leishmaniasis (CL) in the Venezuelan Andean Region / Bionomía de tres especies de flebótomos vectores (Diptera: Psychodidae) en El Carrizal, un foco endémico de leishmaniasis cutánea en la región Andina Venezolana

We studied some aspects of the bionomics of phlebotomine vectors of cutaneous leishmaniasis (CL) at El Carrizal, Municipality Tovar, Venezuela: Lutzomyia youngi, Lutzomyia spinicrassa and Lutzomyia migonei. Monthly catches were performed throughout 2008 using CDC and Shannon traps. We investigated: (i) species richness, (ii) relationships richness-traps-habitats (domestic, peri-domestic and forest), (iii) monthly fluctuations and relationship with climatic variables, (iv) relationship climatic factors-cutaneous leishmaniasis cases at El Carrizal and Municipality Tovar (2000-2008). Females in the Verrucarum group and males of Lu. youngi prevailed. Catches with CDC revealed the predominance of Lu. migonei outdoors where probably they maintains cutaneous leishmaniasis transmission, while females in the Verrucarum group would contribute to maintain it indoors. The greatest abundance of the three species occurred in November after the heaviest peak of rains. The correlation species-climate for Lu. migonei indicated that temperature seems a limiting factor and precipitation a favouring factor, which agreed with results obtained through Ecological Niche Modelling of this species over 23 records at national level. The correlation cutaneous leishmaniasis incidence-climate showed that the greatest number of cases tends to occur, with a month lag, in the months of most rainfall. These data may support the Ministry of Health, since they provide robust information about "when", "where" and "how" to implement target phlebotomine vectors control methods in the Andean Region of Venezuela.

Se estudiaron algunos aspectos de la bionomía de vectores de leishmaniasis cutánea en la localidad El Carrizal, Municipio Tovar, Venezuela: Lutzomyia youngi, Lutzomyia spinicrassa y Lutzomyia migonei. Las capturas mensuales se realizaron a lo largo del año 2008 usando trampas CDC y Shannon. Se investigó: (i) riqueza de especies, (ii) relaciones riqueza-trampa-hábitat (doméstico, peri-doméstico y selvático), (iii) fluctuaciones mensuales y relación con variables climáticas, (iv) relación factores climáticos-casos de leishmaniasis cutánea en El Carrizal y Tovar (2000-2008). Predominaron las hembras del grupo Verrucarum y los machos de Lu. youngi. Las capturas con CDC revelaron el predominio de Lu. migonei en el habitat peridoméstico donde probablemente mantienen la transmisión, mientras que las hembras del grupo Verrucarum contribuirían a mantenerla dentro de las casas. La mayor abundancia de las tres especies ocurrió en noviembre después del pico más alto de lluvias. La correlación especie-clima para Lu. migonei indicó que la temperatura parece ser un factor limitante y la precipitación un factor favorecedor, lo cual coincidió con los resultados obtenidos a través de la Modelación de Nichos Ecológicóos de esta especie sobre 23 registros a nivel nacional. La correlación de incidencia de leishmaniasis cutánea-clima mostró que el mayor número de casos tiende a ocurrir, con un mes de retraso, en los meses de mayor precipitación. Estos datos pueden ser utiles al Ministerio de Salud ya que proveen información sólida sobre "cuándo", "dónde" y "cómo" implementar métodos de control de vectores en la Región Andina de Venezuela.

Risk factors for Trypanosoma cruzi human infection in Barinas State, Venezuela.

This study attempted to quantify the transmission of Trypanosoma cruzi in children less than 15 years of age in Barinas State, Venezuela and investigate risk factors for infection. Among 3,296 children, 4 (0.12%) were seropositive. The mother of one child also was also seropositive, which suggested that congenital transmission is a possible risk factor for Chagas disease in this area. Seroprevalence among the dwellers of 10 localities was 3.3%. Rhodnius prolixus was detected in 7 localities and in 8% of 125 dwellings. A multivariate logistic regression model showed that infection was associated with age, a dirt floor, and distance from houses to palm trees. The risk of infection is increased by the presence of adventitious sylvatic R. prolixus and transient or residual colonies. Insecticide spraying does not seem justified in this scenario, a finding that was also observed in other Latin American countries. New methods are therefore needed for Chagas disease control programs.

Comparación de métodos de captura intradoméstica de triatominos vectores de la enfermedad de Chagas en Venezuela / Comparison of methods of intradomestic capture of Chagas disease vectors in Venezuela

La estratificación de riesgo para priorizar las acciones de control en función de optimización de recursos, es de gran importancia dentro de las actividades de vigilancia epidemiológica para el control de la Enfermedad de Chagas. Para esto es necesario contar con un método para la detección de la infestación intradomiciliaria por triatominos vectores, que sea de alta sensibilidad, especialmente encondiciones de baja densidad. En este estudio se compararon los resultados obtenidos con 5 métodos de capturas de vectores, 2 de búsqueda pasiva: caja Gómez-Nuñez y papel A4 Bond colocados en el dormitorio principal de las casas, 2 de búsqueda activa por personal técnico especializado: captura directa y captura con un excitante (cypermetrina+tetrametrina) durante ½ hora y por último la recolección efectuada durante un mes por los habitantes, oportunamente entrenados. El estudio se realizó en 11 localidades y 200 casas del Estado Portuguesa y 29 localidades y 350 casas del Estado Barinas. No se encontraron diferencias estadísticamente significativas entre los resultados obtenidos con los diferentes métodos en los dos estados. Sobre el total de las 550 casas exploradas, 132 fueron positivas: en 131 (99,2%) la presencia de triatominos fue detectada por lo habitantes vs. 5,3% por búsqueda activa; 3,38 por la caja Gómez-Nuñez; 3,03% por búsqueda con excitante y 0 por ciento por la hoja de papel. Se colectaron en total 5 especies y 563 individuos. Rhodnius prolixus fue la especie predominante seguido por Triatoma maculata, Panstrongylus geniculatus y R. pictipes.

Risk of Trypanosoma cruzi I (Kinetoplastida: Trypanosomatidae) transmission by Panstrongylus geniculatus (Hemiptera: Reduviidae) in Caracas (Metropolitan District) and neighboring States, Venezuela.

The collection of Panstrongylus geniculatus bugs by inhabitants of dwellings in Caracas city (Metropolitan District) and in the neighboring Miranda and Vargas Sates, Venezuela, allowed for the gathering of data on the potential role of this sylvatic triatomine bug as a vector of Chagas disease in this area. The natural infection by Trypanosoma cruzi was recorded by examining fresh and stained faeces of the bugs. Additionally, a random amplification of polymorphic DNA technique for parasite identification and group typing was employed. A dot-ELISA test was used to identify the gut content of the triatomine bugs with the aim of assessing and quantifying the vector-human contact. Sixty-seven specimens (76.1%) were positive to T. cruzi (identified as T. cruzi I) and 60.2% (53/88) gave a positive reaction to the human antiserum. The human blood-positive samples included mixed blood meals with domestic animals (dog, pig and cow) (9.4%) and with mouse (3.8%). The overall Human Blood Index, measured as the percentage of bugs whose gut contents reacted with human antiserum on the total numbers of bugs that reacted with all the antisera tested, was 98.1%. Almost 41% of the bugs that had fed on humans were also positive for T. cruzi. These data show that the feeding of P. geniculatus on humans does not seem to be accidental and that its rate of infection by T. cruzi is high in this area which is not regarded as endemic for Chagas disease by the National Control Programme. This situation is particularly striking because it occurs in and around Caracas, the capital city, where 20% of the whole population of Venezuela live, human migrations from endemic areas are continuous, people in the crowded shantytown as well as people living in high-quality country houses are equally at risk and the epidemiological cycle Didelphis marsupialis/Rattus rattus-P. geniculatus-human does appear to occur successfully.