A novel phylogeny for the genus Echinococcus, based on nuclear data, challenges relationships based on mitochondrial evidence.

The taxonomic status of Echinococcus, an important zoonotic cestode genus, has remained controversial, despite numerous attempts to revise it. Although mitochondrial DNA (mtDNA) has been the source of markers of choice for reconstructing the phylogeny of the genus, results derived from mtDNA have led to significant inconsistencies with earlier species classifications based on phenotypic analysis. Here, we used nuclear DNA markers to test the phylogenic relationships of members of the genus Echinococcus. The analysis of sequence data for 5 nuclear genes revealed a significantly different phylogeny for Echinococcus from that proposed on the basis of mitochondrial DNA sequence data, but was in agreement with earlier species classifications. The most notable results from the nuclear phylogeny were (1) E. multilocularis was placed as basal taxon, (2) all genotypes of Echinococcus granulosus grouped as a monophyletic entity, and (3) genotypes G8 and G10 clustered together. We conclude that the analysis of nuclear DNA data provides a more reliable means of inferring phylogenetic relationships within Echinococcus than mtDNA and suggest that mtDNA should not be used as the sole source of markers in future studies where the goal is to reconstruct a phylogeny that does not only reflect a maternal lineage, but aims to describe the evolutionary history at species level or higher.

Seasonal fluctuations in prevalence of dog intestinal parasites in public squares of Mar del Plata city, Argentina and its risk for humans / Estacionalidad en la prevalencia de parásitos intestinales caninos en plazas públicas de Mar del Plata, Argentina, y su riesgo para la salud humana

The aim of this work is to analyze whether there is a seasonal prevalence of parasites in dog feces from public squares in Mar del Plata city, Argentina, and to evaluate the climatic conditions that promote the development of parasites and allow the transmission to people. The study was performed in 21 squares from June 2001 to May 2002. Samples were processed by the Willis technique. Differences in prevalence of parasites were examined for significance by the Chisquare test. Climatic data were obtained from the Library of the National Meteorological Service of Argentina. Total parasitic prevalence was higher in winter than in summer. The prevalence of Ancylostoma spp. was higher in the summer-autumn period. For Toxocara canis, the prevalence was higher in winter whereas for Trichuris vulpis, it was higher in winter, spring and summer. This work shows high prevalence of total parasites throughout the year. For Ancylostoma spp., summer and autumn might be the seasons with higher sanitary risk. On the other hand, T. canis could present the higher risk to people in winter and, T. vulpis, would be transmitted throughout the whole year. The seasonal variation in prevalence of dog parasites results in continuous exposure to people visiting the squares, not only Mar del Plata residents but also tourists from other regions of Argentina and the world, with at least one species of parasite with sanitary risk.

El objetivo de este trabajo es analizar la variación estacional de la prevalencia de los parásitos presentes en las heces caninas halladas en plazas públicas de la ciudad de Mar del Plata y evaluar aquellas condiciones climáticas que promueven su desarrollo y permiten su transmisión a la gente. El estudio se realizó en 21 plazas desde junio de 2001 hasta mayo de 2002. Las muestras fueron procesadas con la técnica de Willis. Se utilizó el test de c² para comparar la prevalencia de los diferentes parásitos. Los datos climáticos fueron obtenidos en la biblioteca del Servicio Meteorológico Nacional. La prevalencia total de parásitos fue considerablemente más alta en invierno que en verano. Ancylostoma spp. fue la especie prevalente en el período otoño-verano. La incidencia de Toxocara canis fue más alta en invierno que en el resto de las estaciones, y la de Trichuris vulpis fue alta en invierno, primavera y verano. Este trabajo muestra alta prevalencia de parásitos totales a lo largo de todo el año en las plazas públicas de Mar del Plata. Para Ancylostoma spp. verano y otoño podrían ser las estaciones con riesgo sanitario más alto para la gente. Por otra parte, T. canis podría presentar mayor riesgo para la gente en invierno y T. vulpis sería transmitido durante todo el año. La variación estacional en la prevalencia de los distintos parásitos caninos, con al menos una especie con riesgo sanitario, hace que la exposición para la gente que visita las plazas sea continua e involucre no sólo a la gente que vive en Mar del Plata sino también turistas de otras regiones de Argentina y el mundo.

In vitro and in vivo effects of flubendazole on Echinococcus granulosus metacestodes.

The aim of the present work was to determine the efficacy of flubendazole (FLBZ) against Echinococcus granulosus metacestodes by using in vitro and in vivo models. Groups of 50 microcysts developed in vitro, and groups of 10 peritoneal cysts were obtained from Balb C mice with experimental secondary infections of 8 months. The cysts were placed in Leighton tubes containing 10 ml of culture medium. FLBZ was added to the medium resulting in final concentrations of 5 and 1 microg/ml for mycrocysts treatment and 10, 5, and 1 microg/ml for murine cysts treatment. In vivo treatment was performed on 20 mice that developed an experimental secondary hydatid disease over a period of 11 months. FLBZ was given (1.5 mg/kg) by the oral route once a day for 50 days. A loss of turgidity was detected in all in vitro drug treated cysts irrespective of the drug concentration or parasite origin. Inspection of treated cysts by scanning electron microscopy (SEM) revealed that the germinal layer lost it characteristic multicelular structure. These results were confirmed on the ultrastructural level by transmission electron microscopy (TEM), treated metacestodes had undergone considerable degenerative changes after the in vitro treatment. The results obtained after the in vivo treatment with FLBZ showed no significant difference between the control and treated groups related to the weight of cyst masses. However, the ultrastructural study at TEM of cysts that developed in mice from the treated group revealed alterations in the germinal layer with the presence of numerous vacuoles. With regard to the ultrastructural study at SEM, only cellular debris of the germinal layer could be seen. In conclusion, the data obtained clearly demonstrate that in vitro and in vivo treatment with FLBZ is effective against E. granulosus metacestodes.

Seasonal fluctuations in prevalence of dog intestinal parasites in public squares of Mar del Plata city, Argentina and its risk for humans.

The aim of this work is to analyze whether there is a seasonal prevalence of parasites in dog feces from public squares in Mar del Plata city, Argentina, and to evaluate the climatic conditions that promote the development of parasites and allow the transmission to people. The study was performed in 21 squares from June 2001 to May 2002. Samples were processed by the Willis technique. Differences in prevalence of parasites were examined for significance by the Chi-square test. Climatic data were obtained from the Library of the National Meteorological Service of Argentina. Total parasitic prevalence was higher in winter than in summer. The prevalence of Ancylostoma spp. was higher in the summer-autumn period. For Toxocara canis, the prevalence was higher in winter whereas for Trichuris vulpis, it was higher in winter, spring and summer. This work shows high prevalence of total parasites throughout the year. For Ancylostoma spp., summer and autumn might be the seasons with higher sanitary risk. On the other hand, T. canis could present the higher risk to people in winter and, T. vulpis, would be transmitted throughout the whole year. The seasonal variation in prevalence of dog parasites results in continuous exposure to people visiting the squares, not only Mar del Plata residents but also tourists from other regions of Argentina and the world, with at least one species of parasite with sanitary risk.