BACKGROUND It was previously demonstrated that CMC-20, a nitazoxanide and N-methyl-1H-benzimidazole hybrid molecule, had higher in vitro activity against Giardia intestinalis WB strain than metronidazole and albendazole and similar to nitazoxanide. OBJETIVES To evaluate the in vitro activity of CMC-20 against G. intestinalis strains with different susceptibility/resistance to albendazole and nitazoxanide and evaluate its effect on the distribution of parasite cytoskeletal proteins and its in vivo giardicidal activity. METHODS CMC-20 activity was tested against two isolates from patients with chronic and acute giardiasis, an experimentally induced albendazole resistant strain and a nitazoxanide resistant clinical isolate. CMC-20 effect on the distribution of parasite cytoskeletal proteins was analysed by indirect immunofluorescence and its activity was evaluated in a murine model of giardiasis. FINDINGS CMC-20 showed broad activity against susceptible and resistant strains to albendazole and nitaxozanide. It affected the parasite microtubule reservoir and triggered the parasite encystation. In this process, alpha-7.2 giardin co-localised with CWP-1 protein. CMC-20 reduced the infection time and cyst load in feces of G. muris infected mice similar to albendazole. MAIN CONCLUSIONS The in vitro and in vivo giardicidal activity of CMC-20 suggests its potential use in the treatment of giardiasis.

Abstract INTRODUCTION: Different blood meal sources can affect biology of triatomines. METHODS: We studied the influence of two different blood meal sources (rabbits and hens) on five biological parameters in Meccus phyllosomus pallidipennis and collected biological parameters. RESULTS: The cohort fed on rabbits had a shorter median life-cycle and lower mortality rate than that fed on hens. Both cohorts required a similar number of blood meals to reach adulthood. Median longevity and number of blood meals for adults were similar between both cohorts. CONCLUSIONS The studied parameters reflect the high grade of adaptation of M. p. pallidipennis feeding on different hosts.

Resumen Más de la mitad de la población humana está expuesta a contraer infecciones transmitidas por mosquitos. El cambio climático y la aparición de cepas resistentes a los insecticidas tradicionalmente utilizados han motivado la búsqueda de nuevos agentes capaces de controlar las poblaciones de mosquitos. Los aceites esenciales han resultado ser eficaces agentes repelentes y larvicidas. El objetivo de este trabajo fue revisar las investigaciones llevadas a cabo en los últimos años sobre la actividad larvicida de los aceites esenciales y sus componentes contra mosquitos de los géneros Aedes, Anopheles y Culex, así como los últimos reportes sobre su posible mecanismo de acción.

Abstract More than half of the human population is exposed to mosquito-borne infections. Climate change and the emergence of strains resistant to traditionally used insecticides have motivated the search of new agents for mosquito population control. Essential oils have been effective repellents and larvicidal agents. The aim of this work was to review research studies conducted in recent years on the larvicidal activity of essential oils and their components against Aedes, Anopheles and Culex mosquitoes, as well as the latest reports about their possible mechanism of action.

Trypanosoma cruzi has a particular cytoskeleton that consists of a subpellicular network of microtubules and actin microfilaments. Therefore, it is an excellent target for the development of new anti-parasitic drugs. Benzimidazole 2-carbamates, a class of well-known broad-spectrum anthelmintics, have been shown to inhibit the in vitro growth of many protozoa. Therefore, to find efficient anti-trypanosomal (trypanocidal) drugs, our group has designed and synthesised several benzimidazole derivatives. One, named JVG9 (5-chloro-1H-benzimidazole-2-thiol), has been found to be effective against T. cruzi bloodstream trypomastigotes under both in vitro and in vivo conditions. Here, we present the in vitro effects observed by laser scanning confocal and scanning electron microscopy on T. cruzi trypomastigotes. Changes in the surface and the distribution of the cytoskeletal proteins are consistent with the hypothesis that the trypanocidal activity of JVG9 involves the cytoskeleton as a target.

The values of biological parameters related to the life cycles of four populations of Meccus longipennis (Reduviidae: Triatominae) were evaluated. Cohorts of each of the four studied populations from different geographical areas of Mexico were maintained under similar laboratory conditions and then compared. The population from El Saucito de Araujo was different from the other three studied populations, which could help explain the secondary importance of M. longipennis in the state of Chihuahua. This paper also supports the proposition that biological traits are important criteria for determining relationships between populations.

The values of biological parameters related to hatching, lifespan, the number of blood meals between moults, mortality, time lapse before the beginning of feeding, feeding time and defecation delay for each instar of three Mexican-American species of Triatominae, Triatoma recurva, Triatoma protracta (former subspecies protracta) and Triatoma rubida (former subspecies uhleri), were evaluated and compared. No significant (p > 0.05) differences were recorded among the three species with respect to the average time required to hatch. This time was approximately 19 days. The average egg-to-adult development time was significantly (p < 0.05) shorter for T. rubida. The number of blood meals at each nymphal instar varied from one-five for each species. The mortality rates were higher for the first-instar nymphs of the three species studied. The mean time lapse before the beginning of feeding was between 0.3-3 min for most nymphs of all instars of each species studied. The mean feeding time was the longest for T. recurva, followed by T. protracta. The defecation delay was less than 10 min for T. recurva and T. rubida. Given these results, only T. rubida should be considered an important potential vector of Trypanosoma cruzi transmission to humans in areas of Mexico where these species exist, whereas T. recurva and T. protracta would be of secondary importance.

Se reporta por primera vez la recolecta de ejemplares de Meccus longipennis (Usinger) y de Triatoma recurva (Stål) en el estado de Durango. La búsqueda de triatominos se realizó durante 12 meses (septiembre 2010-agosto 2011) en cuatro comunidades de dos municipios (Pueblo Nuevo y Mezquital) del estado de Durango. Se recolectaron 71 ejemplares de M. longipennis y seis de T. recurva provenientes mayoritariamente del interior de las viviendas humanas (recámaras). La presencia de M. longipennis en el área abre la posibilidad de un riesgo potencial de transmisión de Trypanosoma cruzi Chagas a las poblaciones humanas del área estudiada.

The collection of Meccus longipennis (Usinger) and of Triatoma recurva (Stål) in the state of Durango is reported for the first time. Both species were collected during twelve months (from September 2010 to August 2011) in four localities of two municipalities (Pueblo Nuevo y Mezquital) in the state of Durango. Seventy one specimens of M. longipennis and six of T. recurva were collected mostly from indoors of human dwellings (bedrooms). Presence of M. longipennis means a potential risk of transmission of Trypanosoma cruzi Chagas to human populations in the study area.

An entomological study of triatomine species was carried out to assess their prevalence in 10 localities of the state of Michoacán, Mexico. Entomological indices were calculated to estimate the risk for vector-borne transmission of Trypanosoma cruzi to the human population in this area. Four triatomine species (Triatoma barberi, Triatoma dimidiata, Meccus pallidipennis and Meccus longipennis) were collected from the study area. This is the first report of M. longipennis and T. dimidiata in Michoacán. M. pallidipennis was significantly (p < 0.05) more abundant than any of the other species collected in the study area. Infection indices were greater than 50% for each of the four collected triatomine species. Significantly more triatomines were collected from intradomiciliary areas than from peridomiciliary or sylvatic areas. Infestation, crowding and density indices were low, whereas colonisation indices were high in five localities. The current vectorial conditions in the study area require continuous entomological and serological surveillance to diminish the risk of T. cruzi transmission to human populations.

The degree of reproductive isolation between Meccus phyllosomus and the remaining five species of the genus Meccus, as well as between Meccus bassolsae and Meccus pallidipennis, Meccus longipennis and Meccus picturatus, was examined. Fertility and the segregation of morphological characteristics were examined in two generations of hybrids from crosses between these species. The percentage of couples with offspring (fertile) was high in the vast majority of sets of crosses, with the exception of that between ♀M. phyllosomus and ♂Meccus mazzottii. In sets of crosses involving M. bassolsae specimens, no first-generation (F1) individuals were morphologically similar to M. bassolsae, but instead shared the morphology of the other parental species. A similar phenomenon was observed in most sets of crosses involving M. phyllosomus. These results indicated that different degrees of reproductive isolation exist among the species of Meccus involved in this study. The biological evidence obtained in this study does not support the proposal that M. bassolsae is a full species. It could indicate that, on the contrary, it should be considered a subspecies of a single polytypic species. The biological evidence does support the proposal that M. phyllosomus is a full species.

En este estudio se reportan las colectas de triatominos realizadas en los estados mexicanos de Chihuahua y Aguascalientes, de los cuales se carecía de datos recientes sobre presencia e infección por Trypanosoma cruzi en triatominos vectores de la enfermedad de Chagas. En Chihuahua se colectaron exclusivamente ejemplares adultos de Triatoma recurva y de Meccus phyllosomus longipennis, con porcentajes de infección por Trypanosoma cruzi de 5,17% y 12,5%, respectivamente. En Aguascalientes se colectaron ejemplares adultos y ninfas sólo de M. p. longipennis, con un porcentaje de infección por T. cruzi de 96,7%. Los resultados indican nula colonización de la vivienda por triatominos en el área de estudio en Chihuahua, así como bajo riesgo de infección por T. cruzi. En contraste, en el área de estudio en Aguascalientes se prevé riesgo alto de infección por T. cruzi a la población. Se requiere de investigaciones entomológicas adicionales en las áreas de estudio para clarificar el riesgo de transmisión vectorial de T. cruzi a los humanos y reservorios animales.

On this study are reported some collections of triatomines from the Mexican states of Chihuahua and Aguascalientes, where recent data about vector infection by Trypanosoma cruzi was not available. In the state of Chihuahua only adult specimens of Triatoma recurva and of Meccus phyllosomus longipennis were collected. Percentages of infection by T. cruzi were respectively 5.17% and 12.5%. In the state of Aguascalientes adult as well nymph specimens of M. p. longipennis were collected. Percentage of infection by T. cruzi reached 96.7%. Our results show a null colonization of human dwellings by triatomines in the study area of the state of Chihuahua, as well as low risk of infection by T. cruzi for the inhabitants of that area. On the other hand, in the study area in the state of Aguascalientes, the risk of infection by T. cruzi was high for inhabitants of the area. Complementary entomological studies are necessary to establish the proper risk of infection by vectorial transmission of T. cruzi to human and animal reservoir host in the two studied areas.

Introducción. En México, la enfermedad de Chagas es un problema de salud pública. Se han registrado, al menos, 30 especies de triatominos vectores, la mayoría con hábitos domésticos y tendencia a invadir las viviendas humanas. Objetivo. Evaluar la utilidad de gallineros en la vigilancia de la presencia de triatominos peridomésticos y conocer su situación en el área de estudio. Materiales y métodos. Se realizó un estudio de campo, experimental, longitudinal, por un año. Se instalaron 50 gallineros en 50 viviendas de tres localidades del municipio de Teocuitatlán de Corona, en el estado de Jalisco, México. Se inspeccionaron los gallineros y las viviendas para saber si aquéllos podían detectar la presencia de triatominos, para estimar la situación vectorial en el área. Resultados. Se recolectaron significativamente (p<0,05) más especímenes de Meccus longipennis (93%) que de Triatoma barberi (7%). Aproximadamente, un tercio (31,3%) de los especímenes de M. longipennis y ninguno de T. barberi se recolectó en los gallineros. M. longipennis tuvo un porcentaje de infección por Trypanosoma cruzi de 25,7%, y T. barberi, de 53,2%. Los meses de primavera y verano fueron en los que se recolectaron más ejemplares de triatominos. Conclusiones. Los gallineros se consideran una herramienta útil para detectar la presencia de ejemplares de M. longipennis, aunque no funcionaron para T. barberi. Se confirma la importancia de M. longipennis y se establece el posible rol primario de T. barberi como vector en el área de estudio.

Introduction. Chagas disease is an important public health problem in Mexico. At least 30 Triatominae species, vectors of that disease have been recorded, most with domiciliary habits and tendency to invade human dwellings. Objective. The usefulness of using chicken coops was evaluated for monitoring peridomestic triatomine presence, identity and infection with Trypanosoma cruzi. Materials and methods. A year-long, longitudinal field study was designed. Fifty chicken coops were placed in 50 human dwellings on three localities in Teocuitatlán de Corona County, Jalisco State, Mexico. The coops and human dwellings were routinely inspected to determine if the coops alone can monitor for the presence of triatomines. Results. Significantly (p<0.05) more Meccus longipennis specimens (93.0%) than of Triatoma barberi (7.0%) were collected. Almost a third (31.3%) of M. longipennis and none of T. barberi specimens were collected in the coops. Meccus longipennis was found with infections of Trypanosoma cruzi of 25.7%, and T. barberi of 53.2%. Most triatomines were collected in spring and summer months. Conclusions. Chicken coops proved useful tools to detect presence of M. longipennis; however, they were not effective for T. barberi. The importance of M. longipennis as a potential vector was confirmed, and the possible primary vector role of T. barberi was established as well.

In seven studied communities of Western Mexico, triatomine specimens were sympatrically collected, some with atypical morphological characteristics in contrast to pure specimens, which were presumed to be hybrids. More than 200 specimens of Meccus pallidipennis and Meccus longipennis with brown-yellow markings on dorsal connexival segments were collected in Ahuacapán and Quitupan. In La Mesa, more than 60 specimens similar to Meccus picturatus in most morphological characteristics (including size) were collected, although they presented a largely yellowish corium like M. pallidipennis. Interfertility was proven between all of the studied wild hybrid specimens, as well as between all the experimental laboratory hybrids. Two different phenotypes (M. picturatus and M. longipennis) were obtained from crosses between M. picturatus x M. picturatus and M. longipennis x M. longipennis from the three studied localities in state of Nayarit as from La Mesita. Results support the hypothesis that the subspecific ranking of those triatomines may, therefore, be more appropriate because reproductive isolation has not been developed and complete interbreeding was recorded.

Several biological parameters related to the Triatoma mexicana life-cycle were evaluated in this study. Three cohorts were maintained under different combinations of temperature and relative humidity (RH): 25ºC/50% RH; 25ºC/75% RH; and 30ºC/75% RH. Observed hatching rates varied from 49-57.5% whereas the average time of hatching varied from 19.5-22.7 days. In the three cohorts studied, the mean time-lapse between presentation of the blood meal and the beginning of feeding was less than 5 min in all instars; the mean feeding time was longer than 10 min in all the instars; the post-feed defecation delay was over 10 min in all the instars. Less than 50% of nymphs in each cohort completed the cycle and the average time from 1st instar nymph to adult was more than 255 days for the three cohorts. The number of blood meals before molt at each nymphal instar varied from 1-9. Our results appear to indicate a lack of influence of temperature and RH on the biological parameters of T. mexicana that were studied, which could reflect the adaptation capacity of this species. We also conclude that T. mexicana can not be considered an effective transmitter of Trypanosoma cruzi to human populations in areas where this species is currently present.

The present work is a thorough investigation of the degree of reproductive isolation between Meccus mazzottii and Meccus longipennis, Meccus picturatus, Meccus pallidipennis and Meccus bassolsae, as well as between M. longipennis and M. picturatus. We examined fertility and segregation of morphological characteristics in two generations of hybrids derived from crosses between these species. The percentage of pairs with (fertile) offspring was highest in the set of crosses between M. longipennis and M. picturatus, and lowest between M. mazzottii and M. picturatus. Most first-generation (F1) individuals from crosses involving M. mazzottii were morphologically similar to this species, while only F1 x F1 progeny of parental crosses between M. mazzottii and M. longipennis had offspring second generation that looked like M. mazzottii. The results indicate that different degrees of reproductive isolation apparently exist among the species of the Phyllosoma complex examined in this study. The biological evidence obtained in this study does not support the proposal that M. longipennis and M. picturatus are full species. It could indicate on the contrary, that both could be considered as subspecies of a single polytypic species. On the other hand, biological evidence supports the proposal that M. mazzottii is a full species.

Aspects related to hatching, lifetime, number of blood meals for molting, mortality, feeding time, and postfeeding defecation delay were evaluated and compared in each instar of three North American Triatominae: Triatoma gerstaeckeri, Triatoma lecticularia and Triatoma protracta, all of them fed on rabbits. No significant differences (p > 0.05) were found among the three species regarding mean hatching rate, which was close to 20 days. Egg-to-adult development times were significantly shorter (p < 0.05) for T. lecticularia. Number of blood meals for molting to next instar ranged from one to five for T. protracta, and from one to six for T. gerstaeckeri and T. lecticularia. Mortality rates were higher in younger nymphs of T. lecticularia and T. protracta, while rates in T. gerstaeckeri were higher in fifth-instar nymphs. Mean feeding time was longest in T. gerstaeckeri, followed by T. lecticularia. More than twice the number of T. gerstaeckeri nymphs completed the development process, if compared to the nymphs from the other two species. Defecation delay was less than 10 min for T. lecticularia, T. protracta and the youngest nymphs of T. gerstaeckeri. Results point out that these three species may be important potential vectors of Trypanosoma cruzi for human populations, in areas of Mexico where these species are currently present.