Self-fertilization, long-distance flash invasion and biogeography shape the population structure of Pseudosuccinea columella at the worldwide scale.

Population genetic studies are efficient for inferring the invasion history based on a comparison of native and invasive populations, especially when conducted at species scale. An expected outcome in invasive populations is variability loss, and this is especially true in self-fertilizing species. We here focus on the self-fertilizing Pseudosuccinea columella, an invasive hermaphroditic freshwater snail that has greatly expanded its geographic distribution and that acts as intermediate host of Fasciola hepatica, the causative agent of human and veterinary fasciolosis. We evaluated the distribution of genetic diversity at the largest geographic scale analysed to date in this species by surveying 80 populations collected during 16 years from 14 countries, using eight nuclear microsatellites and two mitochondrial genes. As expected, populations from North America, the putative origin area, were strongly structured by selfing and history and harboured much more genetic variability than invasive populations. We found high selfing rates (when it was possible to infer it), none-to-low genetic variability and strong population structure in most invasive populations. Strikingly, we found a unique genotype/haplotype in populations from eight invaded regions sampled all over the world. Moreover, snail populations resistant to infection by the parasite are genetically distinct from susceptible populations. Our results are compatible with repeated introductions in South America and flash worldwide invasion by this unique genotype/haplotype. Our study illustrates the population genetic consequences of biological invasion in a highly selfing species at very large geographic scale. We discuss how such a large-scale flash invasion may affect the spread of fasciolosis.

Real-time PCR strategy for rapid discrimination among main lymnaeid species from Argentina.

Snails of the Family Lymnaeidae act as an intermediate hosts of Fasciola hepatica worldwide. The taxonomy of lymnaeid species is relevant for epidemiological studies and molecular strategies are increasingly used for that purpose. This work presents the first report of a real-time PCR approach used to identify the most important lymnaeid species in the Southern Cone of South America. Species discrimination is based on the sequence polymorphism located within the helix E10-1 of the variable region V2 of the 18S rRNA genes, which yields amplicons with clearly different melting temperatures. This procedure minimises the risk of carry-over contamination because it does not require post-PCR manipulations, and the whole protocol can be completed in less than 4h with a single snail foot as starting material. This method was successfully carried out in a blind study that included a panel of 20 Galba truncatula, 5 Lymnaea viatrix, 5 Lymnaea diaphana and 5 Pseudosuccinea columella specimens from different endemic areas for fasciolosis. This molecular approach constitutes a key laboratory tool complementing ecological studies that ultimately will promote more efficient control strategies.

Comparison of two coprological methods for the veterinary diagnosis of fasciolosis

Compararam-se a sensibilidade dos métodos padrão de sedimentação fecal (MPSF) e modificado da tamisação e coloração das fezes (MTCF)-quatro tamises, ambos empregados no diagnóstico de infecção por Fasciola hepatica. Amostras de fezes foram obtidas de 51 bovinos oriundos de uma área endêmica por fasciolose no Sudoeste da Argentina. Cada amostra foi colocada em um recipiente contendo formalina 5%. Oito mililitros da suspensão, equivalente a 2g de fezes, foram usados em cada método testado, registrando-se o número de ovos por amostra. A porcentagem de amostras positivas pelo MTCF (27/51) foi maior que a apresentada pelo MPSF (11/51), o que representou 60% de amostras falso-negativas. A porcentagem de concordância pelos dois métodos foi 41%. A complexidade do MPSF pode diminuir sua sensibilidade pela perda de ovos durante o processamento. Estes resultados confirmaram que o MPSF subestima a contagem de ovos e que o método MTCF é mais confiável.

Epidemiología de la enfermedad de Chagas en San Roque, Corrientes: infestación por triatominos y seroprevalencia humana / Epidemiology of Chagas disease in San Roque, Corrientes: Triatomine infestation and human seroprevalence

El objetivo fue evaluar la presencia de triatominos en ecotopos domésticos y extradomésticos localizados en el área circundante a una comunidad de palmas, determinar el índice de infección de los triatominos y estimar la prevalencia humana de individuos serorreactivos al Trypanosoma cruzi. Los muestreos se efectuaron en primavera de 1998 y en verano de 1999 en Colonia Laurel, Departamento San Roque, Corrientes, Argentina. La colección de triatominos se realizó por el método de captura/ hora/ hombre. La identificación del T. cruzi se efectuó según criterios morfológicos y por PCR. El diagnóstico de la infección chagásica se llevó a cabo en pobladores voluntarios mediante las pruebas de hemaglutinación indirecta e inmunofluorescencia indirecta. Se analizaron 16 viviendas, 11 resultaron infestadas por Triatoma infestans y dos por Triatoma sordida. Se colectaron 122 ejemplares de T. infestans de todas las edades. La infección por T. cruzi fue 16.7%. Triatoma infestans fue la especie predominante en el domicilio y T. sordida en el peridomicilio. El 77.9% (88/113) de los pobladores se analizaron serológicamente y la prevalencia fue 26.1%. La tasa de infección fue relativamente alta en el grupo etario de 0-10 años. El porcentaje de seropositividad se incrementó con la edad y ascendió al 66.6% en los pobladores de más de 40 años. La presencia de T. infestans, de pobladores serorreactivos y de niños seropositivos ≤4 años, así como las condiciones precarias de vida, indican que la endemia chagásica continúa siendo un problema en el área de estudio.

Estudio del foco en un caso de fasciolosis humana en Neuquén / A focus study from a case of human fascioliasis in Neuquén

El objetivo del trabajo fue realizar un estudio de foco relacionado con un caso de fasciolosis humana ocurrido en abril de 2002 en Loncopué, Neuquén, Argentina. La confirmación diagnóstica se efectuó en mayo de 2002 por el test de ELISA. En noviembre de 2002 se realizó un muestreo en el área rural donde se ubicaba la vivienda de la paciente, ya restablecida, y se le tomó una nueva muestra de sangre. El suero de la paciente continuó reactivo para antígenos de Fasciola hepatica. Se muestrearon plantas de berro para detectar metacercarias. Se recolectaron caracoles en cuatro canales de riego conectados a un canal principal. Los caracoles fueron trasladados vivos para su identificación, medición y examen de infección. Se recolectaron 35 muestras fecales de ganado de cría. No se observaron metacercarias en las hojas de berro examinadas (n=222). Se recolectaron 130 caracoles identificados como Lymnaea viatrix y 2 de 101 ejemplares (2%) estaban infectados con larvas de F. hepatica. Las prevalencias en el ganado adulto fueron: 100% (10/10) para caprinos, 82% (9/11) para ovinos y 86% (6/7) para bovinos. El número de huevos eliminados por las cabras (mediana = 20.7; Q1=6.2; Q3=34.5) y ovejas (4, 18, 13) infectadas, resultó mayor que el eliminado por vacas (0.3; 0.3; 1.7) (p<0.01). La práctica de control local no tuvo efecto aparente en este caso, por lo que deberían revisarse los calendarios de tratamiento y los antiparasitarios utilizados. Los resultados muestran que el ganado criado a pequeña escala por los pobladores debe incluirse en los programas de control. Se discute la posible importancia de la fasciolosis humana en Argentina.

Estudio del foco en un caso de fasciolosis humana en Neuquén / A focus study from a case of human fascioliasis in Neuquén

El objetivo del trabajo fue realizar un estudio de foco relacionado con un caso de fasciolosis humana ocurrido en abril de 2002 en Loncopué, Neuquén, Argentina. La confirmación diagnóstica se efectuó en mayo de 2002 por el test de ELISA. En noviembre de 2002 se realizó un muestreo en el área rural donde se ubicaba la vivienda de la paciente, ya restablecida, y se le tomó una nueva muestra de sangre. El suero de la paciente continuó reactivo para antígenos de Fasciola hepatica. Se muestrearon plantas de berro para detectar metacercarias. Se recolectaron caracoles en cuatro canales de riego conectados a un canal principal. Los caracoles fueron trasladados vivos para su identificación, medición y examen de infección. Se recolectaron 35 muestras fecales de ganado de cría. No se observaron metacercarias en las hojas de berro examinadas (n=222). Se recolectaron 130 caracoles identificados como Lymnaea viatrix y 2 de 101 ejemplares (2%) estaban infectados con larvas de F. hepatica. Las prevalencias en el ganado adulto fueron: 100% (10/10) para caprinos, 82% (9/11) para ovinos y 86% (6/7) para bovinos. El número de huevos eliminados por las cabras (mediana = 20.7; Q1=6.2; Q3=34.5) y ovejas (4, 18, 13) infectadas, resultó mayor que el eliminado por vacas (0.3; 0.3; 1.7) (p<0.01). La práctica de control local no tuvo efecto aparente en este caso, por lo que deberían revisarse los calendarios de tratamiento y los antiparasitarios utilizados. Los resultados muestran que el ganado criado a pequeña escala por los pobladores debe incluirse en los programas de control. Se discute la posible importancia de la fasciolosis humana en Argentina. (AU)

Epidemiología de la enfermedad de Chagas en San Roque, Corrientes: infestación por triatominos y seroprevalencia humana / Epidemiology of Chagas disease in San Roque, Corrientes: Triatomine infestation and human seroprevalence

El objetivo fue evaluar la presencia de triatominos en ecotopos domésticos y extradomésticos localizados en el área circundante a una comunidad de palmas, determinar el índice de infección de los triatominos y estimar la prevalencia humana de individuos serorreactivos al Trypanosoma cruzi. Los muestreos se efectuaron en primavera de 1998 y en verano de 1999 en Colonia Laurel, Departamento San Roque, Corrientes, Argentina. La colección de triatominos se realizó por el método de captura/ hora/ hombre. La identificación del T. cruzi se efectuó según criterios morfológicos y por PCR. El diagnóstico de la infección chagásica se llevó a cabo en pobladores voluntarios mediante las pruebas de hemaglutinación indirecta e inmunofluorescencia indirecta. Se analizaron 16 viviendas, 11 resultaron infestadas por Triatoma infestans y dos por Triatoma sordida. Se colectaron 122 ejemplares de T. infestans de todas las edades. La infección por T. cruzi fue 16.7%. Triatoma infestans fue la especie predominante en el domicilio y T. sordida en el peridomicilio. El 77.9% (88/113) de los pobladores se analizaron serológicamente y la prevalencia fue 26.1%. La tasa de infección fue relativamente alta en el grupo etario de 0-10 años. El porcentaje de seropositividad se incrementó con la edad y ascendió al 66.6% en los pobladores de más de 40 años. La presencia de T. infestans, de pobladores serorreactivos y de niños seropositivos ≤4 años, así como las condiciones precarias de vida, indican que la endemia chagásica continúa siendo un problema en el área de estudio. (AU)

Triatoma sordida Stål 1859 (Hemiptera, Reduviidae: Triatominae) in palms of northeastern Argentina.

Several palms species provide an important habitat for triatomines and associate vertebrates in tropical America. The objective of this work is to identify the triatomine species living in the palms of rural areas in the Province of Corrientes, and to estimate the potential epidemiological risk they represent for the residents of nearby houses. The survey was carried out in a palm community in Colonia Laurel, Department San Roque, Province of Corrientes, Argentina. Samplings were performed in October, November and December 1998; January, February and March 1999; May and June 1999. Thirty palms: 27 (90%) Butia yatay (Mart.) Becc. and 3 Acrocomia aculeata (Jacq.) Lodd ex Mart. were dissected. Triatoma sordida Stål 1859 was found in 96.2% of B. yatay and in all the A. aculeata palms. A total of 272 live T. sordida was collected; 36 of them (13.2%) were found in bird nests in the frond and the remainder in other locations of the tree. The mean number of triatomines per palm was 9.6 (range 1-60, mode 2). T. sordida was collected during all the sampling months and all stages were present at all seasons. The highest population density was reached in spring and the lowest in autumn. Trypanosoma cruzi was detected in 38.5% in feces of 174 examined insects and identified as such, both by microscopical examination and PCR. This is the first finding of T. sordida populations in B. yatay, an endemic palm of South America distributed in southern Brazil, Uruguay and northeastern Argentina. The high infection prevalence found in this work suggests that T. sordida plays an essential role in the maintenance of the wild T. cruzi transmission cycle in northeastern Argentina.

Probability of Trypanosoma cruzi transmission by Triatoma infestans (Hemiptera: Reduviidae) to the opossum Didelphis albiventris (Marsupialia: Didelphidae).

The probability of Trypanosoma cruzi transmission to opossums by independent events of predation and fecal contamination during feeding ("biting") with positive Triatoma infestans was estimated. Negative female opossums were challenged for 23 hr with 10 infected third and fourth instars of T. infestans, and tests for positivity for T. cruzi by xenodiagnosis were performed at 30, 60, and 90 days. From these data, seven probability parameters were estimated by maximum likelihood, and likelihood ratio statistics confidence intervals were calculated. Simultaneous estimation of p1 (probability that a "bite" will infect an opossum), p3 (probability that a bug that has been eaten by an opossum will infect it), and p6 (probability that the opossum will become infected if faced with an infected triatomine), resulted in p1 = 0.06, p3 = 0.075, and p6 = 0.059. On average, each opossum should be exposed to an average of 700 encounters with bugs during its life, resulting in about eight potentially infective contacts, to produce the 35% opossum prevalence found in the field.

Characterization of Triatoma guasayana biotopes in a hardwood forest of Santiago del Estero, Argentina.

Triatoma guasayana is a silvatic triatomine species distributed in Argentina, Bolivia and Paraguay. The study was performed in a secondary forest of Santiago del Estero, Argentina. The abundance of T. guasayana was evaluated by census in the following wild biotopes: quimiles (Opuntia quimilo), chaguares (dry bromeliads), logs and underground burrows. Ten biotopes of each type were dismantled in winter (August) and another 40 in summer (January); all fauna was recorded. The biotopes most infested by T. guasayana were quimiles (65%), followed by chaguares (55%), and logs (25%). Quimiles and chaguares were infested in both seasons, whereas logs were positive only in summer and burrows were never infested. Infestation and abundance were higher in summer than in winter. The biotope structure is a key factor for T. guasayana colonization. The larger number of refuges, the constant presence of blood sources and suitable inner microclimatic conditions offered by quimiles may favour the persistence of T. guasayana colonies. The richness of invertebrate fauna per type of biotope was ranked in the same order as that of T. guasayana, suggesting similar microhabitat requirements for all studied arthropods.

Triatoma infestans in Greater Buenos Aires, Argentina.

The Health Administration Agencies of many municipalities in Greater Buenos Aires (GBA) receive frequent reports on triatomines in houses. The aim of this work was to identify and describe the dispersal foci of Triatoma infestans in an urban neighborhood of GBA, and contribute to the knowledge of the epidemiological situation in the region. In June 1998, potentially infested places were entomologically evaluated. T. infestans was only detected in a hen building for egg production, which housed approximately 6,000 birds. A total of 2,930 insects were collected. Density was about 9 triatomines/m(2). The proportions of fifth instar nymphs and adults were significantly higher than those of the other stages (p<0.001). The number of triatomines collected largely exceeded the highest domestic infestation found in one house from rural endemic areas of Argentina. Though triatomines were negative for Trypanosoma cruzi, they could acquire the parasite by coming in contact with infected people living in GBA. Besides, the numerous and widely distributed places housing hens and chickens, would favor the settlement of the vector. Together, both facts may constitute a risk of parasitic vectorial transmission. It is recommended to intensify systematic activities of vector search and case detection in GBA.

Triatoma infestans in Greater Buenos Aires, Argentina

The Health Administration Agencies of many municipalities in Greater Buenos Aires (GBA) receive frequent reports on triatomines in houses. The aim of this work was to identify and describe the dispersal foci of Triatoma infestans in an urban neighborhood of GBA, and contribute to the knowledge of the epidemiological situation in the region. In June 1998, potentially infested places were entomologically evaluated. T. infestans was only detected in a hen building for egg production, which housed approximately 6,000 birds. A total of 2,930 insects were collected. Density was about 9 triatomines/m². The proportions of fifth instar nymphs and adults were significantly higher than those of the other stages (p<0.001). The number of triatomines collected largely exceeded the highest domestic infestation found in one house from rural endemic areas of Argentina. Though triatomines were negative for Trypanosoma cruzi, they could acquire the parasite by coming in contact with infected people living in GBA. Besides, the numerous and widely distributed places housing hens and chickens, would favor the settlement of the vector. Together, both facts may constitute a risk of parasitic vectorial transmission. It is recommended to intensify systematic activities of vector search and case detection in GBA

Interactions between Triatoma infestans and Triatoma sordida (Hemiptera: Reduviidae) in artificial ecotopes: population growth and age structure.

Interaction characteristics between Triatoma infestans Klug, 1834 and Triatoma sordida Stal, 1859 populations were studied in artificial ecotopes for 16 mo. The experimental design involved simultaneous treatments with T. infestans and T. sordida together in the same experimental unit (EU) and each separately in two control units (CU) made of adobe bricks. Chickens were used as host animals. Each unit was dismantled monthly to estimate triatomine population size and age structure, rebuilt, and repopulated with the same insects. In both units, T. infestans population growth followed a logistic model, whereas T. sordida did not show this pattern. T. infestans completed a generation in 24 wk (EU) and 32 wk (CU1), whereas T. sordida did not complete one generation during this period. We concluded that T infestans showed a better colonizing success than T. sordida. After living together for 1 yr, an interference process took place that resulted in the extinction of T. sordida.

Ecogenetics of Triatoma sordida and Triatoma guasayana (Hemiptera: reduviidae) in the Bolivian chaco.

Triatoma guasayana and two putative cryptic species pertaining to T. sordida complex (named groups 1 and 2) occur in sympatry in the Bolivian Chaco. Using multilocus enzyme electrophoresis and subsequent genetic analysis, our work assesses their population distribution and dispersal capacity in domestic, peridomestic, and silvatic environments. Our collections by light trap in the silvatic environment indicated a predominance of T. guasayana and T. sordida group 2 and a lesser abundance of T. sordida group 1 ( pound 10% of the total of captures). Their similar distribution in two silvatic areas 80 km apart supports the hypothesis of their homogeneous dispersal through the Bolivian Chaco. The distribution of T. guasayana and T. sordida groups 1 and 2 was similar between silvatic environment and peridomestic ecotopes where 25% of positive places was occupied by two or three species. Bromeliads were confirmed as favorable shelter for T. guasayana but were free of T. sordida. T. sordida group 1 and to a lesser extent T. guasayana would be more invasive vectors for houses than T. sordida group 2. The spatial partition in the three species sampled in two distant sites suggested a reduced dispersive capacity.

[Prevalence of Trypanosoma cruzi infection in opossum (Didelphis albiventris) in Santiago del Estero, Argentina]. / Estudio de la prevalencia de la infección por Trypanosoma cruzi en zarigüeyas (Didelphis albiventris) en Santiago del Estero, Argentina.

The opossum of the genus Didelphis is one of the principal wild reservoirs of Trypanosoma cruzi and is widely distributed in the Western Hemisphere. Didelphis albiventris is the most common marsupial in Amamá and Trinidad, two communities in the province of Santiago del Estero, Argentina. The D. albiventris population is replaced every year, and the opossum normally has two reproductive periods, one at the beginning of the spring and another at the beginning of the summer. The two litters are weaned, and they leave the mother's marsupial pouch to join the population, the first (G1) at the beginning of the summer and the second (G2) at the beginning of the fall. Between 1988 and 1991 409 D. albiventris opossums were studied, and xenodiagnoses showed that 35% of them were infected with T. cruzi. Annual cycles of renewed infection were observed, with prevalences that ranged between 22% and 43%. The acquisition of the parasite occurred over the entire year, from the summer through the spring. The prevalence of infection increased with age. The G1 individuals tended to present higher prevalences than the G2 individuals, probably from being exposed to transmission for a longer period of time. In the first two (younger) age categories for the opossums, G2 individuals showed higher prevalences than did the G1 individuals. This indicates a significant increase in transmission intensity during the fall. Opossums should be regarded as a potential source of T. cruzi entry to the domestic transmission cycle.

Baja probabilidad de transmisión de trypanosoma cruzi a humanos por triatoma sordida domiciliado en el departamento de Santa Cruz / Low probability of transmission od Trypanosoma cruzi to humans by triatoma sordid domiciled in the department of Santa Cruz

Entre más de 100 especies de triatominae (hemiptera: ruduviidae) descritas en el Nuevo Mundo, triatoma sordida es considerada de significancia epidemiológica como vector de trypanosoma cruzi por su amplia área de distribución y su tendencia para invadir ambientes domésticos.

Comparative meiotic studies in Triatoma sordida (Stål) and T. guasayana Wygodzinsky & Abalos (Reduviidae, Heteroptera).

Triatoma sordida and T. guasayana are competent Trypanosoma cruzi vectors, with overlapping distribution areas in Argentina. Both species are morphologically similar, and their immature stages are hard to discriminate. Cytogenetic studies in the genus Triatoma reveal scarce karyotypic variations, being 2n = 20 + XY the most frequent diploid number in males. In the present work the meiotic behaviour of different Argentinian populations of T. sordida and T. guasayana has been analyzed; the meiotic karyotype of both species has also been compared. The species differ in total chromosome area and in the relative area of the sex chromosomes. These meiotic karyotypic differences constitute an additional tool for the taxonomic characterization of T. sordida and T. guasayana. The analysis of an interpopulation hybrid of T. sordida (Brazil x Argentina) reveals a regular meiotic behaviour; despite the presence of heteromorphic bivalents. Our observations support the hypothesis that karyotype variations through the gain or loss of heterochromatin can not be considered as a primary mechanism of reproductive isolation in Triatoma.

[Leptospira interrogans in a canine population of Greater Buenos Aires: variables associated with seropositivity]. / Leptospira interrogans en una población canina del Gran Buenos Aires: variables asociadas con la seropositividad.

We determined the seroprevalence of leptospirosis in a suburban canine population for the purpose of analyzing the association between different individual and environmental variables and seropositivity for leptospirosis. The study, which was cross-sectional, was performed in July 1992 in a neighborhood of Greater Buenos Aires with approximately 9,500 inhabitants and a canine population of around 2,000 animals. We studied a random sample of 223 dogs and obtained a blood sample from each. Each animal's epidemiologic history was obtained by interviewing the housewife. Serologic measurements were performed by the microagglutination technique with the use of 10 different serotypes of Leptospira interrogans. Of the 223 dogs that were tested, 57% showed seropositivity; 82% of the positive sera coagglutinated with two or more serotypes. The most frequently detected serotypes were canicola and pyrogenes. Seroprevalence in females was less common than in males (P < 0.05) and in puppies less than 1 year old it was less common than in older animals (P < 0.01). Street behavior in the dog and the presence of stagnant water in front of the owner's dwelling were the most important of the risk factors examined. The associations between seropositivity on the one hand and contact with trash deposits, hunting behavior and the presence of rodents inside the dwelling on the other were not statistically significant. Different control measures are discussed.

Spatial distribution of Triatoma guasayana (Hemiptera:Reduviidae) in hardwood forest biotopes in Santiago del Estero, Argentina.

In the study area Triatoma guasayana Wygodzinsky & Abalos is the only wild triatomine found sympatric with Triatoma infestans (Klug) in peridomestic premises. The Trypanosoma cruzi Chagas wild cycle is centered around the same biotopes occupied by T. guasayana, which are also visited mainly by opossums with annual prevalences of 29-50%. Twelve hectares were sampled for 4 consecutive years during all seasons. During that time, 420 T. Guasayana individuals were collected in 11.3% of 1,188 biotopes of 4 types, which included quimiles (the cactus Opuntia quimilo), chaguares (several species of bromeliads), trees, and logs. Quimiles had the highest percentage of positive biotopes (31.5%) followed by chaguares (22.3%), whereas 5% of the logs were found infested. During all seasons, 9.6-15.2% of biotopes were found infested. Distance to artificial biotopes was not statistically significant when comparing the frequency of triatomine numbers per biotope in all biotope-season combinations. With the exception of quimiles in the fall, the mean number of triatomines was higher in chaguares during all seasons. Triatomine abundance by biotope and season strata showed a clumped distribution, except for the quimiles biotope during the summer. When pooling by seasons, the mean number of triatomines in chaguares and quimiles biotope was higher than in logs and trees, with all biotopes showing a strong clumped distribution. When pooling by biotopes, the mean number of T. guasayana was relatively similar for all seasons, with a strong clumped distribution. The strong contagious distribution of T. guasayana in the hardwood forest biotopes may explain the maintenance of the wild cycle of T. cruzi, despite the low number and the low prevalences of the insect vector.