Influence of blood meal source on the biological parameters of Triatoma pallidipennis (Heteroptera: Reduviidae) from Mexico.

Chagas disease is one of the most important vector-borne diseases in Latin America, including Mexico. Triatoma pallidipennis (Stål) (Hemiptera: Reduviidae) is a Mexican triatomine vector commonly associated with different hosts. The influence of six blood meals (rabbits, rats, mice, dogs, cats and chickens) on six biological parameters of the biology of T. pallidipennis was evaluated. A significant difference was found in the period of egg-to-adult development between the five mammalian feeds (mean 195 days) and the chicken feed (221 days). The probability of survival was significantly lower in the chicken cohort (0.285). The total number of blood meals to moult from the first instar to the adult stage was the highest in the chicken cohort (10-15). This cohort had the significantly highest rate of females at the end cycle. The mean number of eggs laid per female and the egg eclosion rate were similar among the six food sources. Most results seemed to be influenced by the higher nutritional quality of the mammalian blood compared to the bird's blood and the increased energy expenditure required for the digestion of bird blood. These results clearly show that T. pallidipennis, unlike other triatomine species, has a high reproductive capacity when feeding on different hosts.

New finding of peridomestic Triatoma infestans (Klug, 1834) (Hemiptera: Reduviidae) in Mexico: Molecular approach using cytochrome B and cytochrome oxidase I.

A previous work morphologically identified some specimens colonizing peridomestic sites of Manzanillo Colima, Mexico, as Triatoma infestans (Klug). In the present study, a molecular approach using cytochrome B and cytochrome oxidase I was implemented for the genetic identification and determination of the origin of that population. Phylogenetic analyses positively identified our studied specimens as belonging to the T. infestans clade based on genetic markers with high posterior probability values, and the haplotype network showed Uruguay, Chile and Argentina as probable countries of origin of the populations in Mexico, which was supported by gene flow and migration index analyses. Due to the proximity of the port of Manzanillo to the collection sites, the introduced specimens were hypothesized to have travelled from the countries of origin to Mexico in a seed shipment inside a TEU (twenty-foot equivalent unit) maritime container. The identification of T. infestans in Mexico represents a serious health problem, and the findings presented here indicate a novel pathway for displacing this vector with the possibility of transmission to any other part of the world, which should be further investigated.

Biological Parameters of Three Populations of Triatoma dimidiata s. s. (Hemiptera: Reduviidae) From Western Mexico.

Chagas disease is a very important vector-borne disease in México, and Triatoma dimidiata sensu stricto (Latreille) is one of the most important vectors of Trypanosoma cruzi Chagas, mainly in southern and central states. In the Pacific Coast states with the highest prevalence of human T. cruzi infection, T. dimidiata s. s. is considered as a secondary vector. However, the vectorial capacity of those populations has not been studied. Therefore, the vector characteristics of three populations of T. dimidiata s. s. in western México were evaluated in this study. The populations were maintained in the laboratory at 27 ± 1°C and 75% ± 5% RH with a 12:12 h (light:dark) regime, fed on rabbits in a fortnight basis. The development times were short (172-238 d), and the number of bloodmeals to molt was low (11). Mortality was moderate (36-45%), the onset of feeding was relatively rapid (0.5-1.7 min), and feedings were extended (>15 min). More than 40% of individuals in most instars defecated in one of three categories: <1 min when feeding (5-37.9%), immediately after feeding (9-28.6%), or in <1 min post feeding (7-25.8%). The median number of laid eggs was high (over 2.5) in the three populations, as were the egg eclosion rates (>86%). Thus, the T. dimidiata s. s. in the three populations are potentially efficient vectors of T. cruzi and could contribute to the high prevalence of infection in human populations in western México.

Behavioral parameters of six populations of Meccus phyllosomus longipennis (Heteroptera: Reduviidae) from areas with high and low prevalences of Trypanosoma cruzi human infection.

Three behaviors of epidemiological importance, namely feeding latency, feeding duration and defecation latency, for six populations of Meccus phyllosomus longipennis (Usinger) from areas of central, western and north-central Mexico with high (HP) and low (LP) prevalence of Trypanosoma cruzi (Chagas) human infection were evaluated in this study. The median feeding latency (the time taken to begin feeding) was highly variable between instars. Within-instar comparisons showed that at least 65% of the LP populations (N3 to adult) started to feed significantly (P < 0.05) later than the HP population, with N1 showing no difference, and N2 from LP populations feeding sooner than those from HP populations. The six populations had similar median feeding durations within instars. A higher (P < 0.05) percentage of the instars from HP populations defecated faster than the respective instars from the three LP populations. Approximately 25% of the young nymphs (N1 to N3) and females in the HP populations defecated < 2 min postfeeding, compared with 4%-6% of the young nymphs and 1.3%-3% of females in the LP populations. Moreover, 17.7%-38.8% of the older nymphs (N4 to N5) in the HP populations and 6.8%-13.4% in the LP populations defecated during or immediately after feeding. Our results indicate that the HP populations have a greater potential than the LP populations to transmit T. cruzi infections, which may underlie the differences in the prevalence of T. cruzi infection in some areas where M. p. longipennis is currently distributed.

Biological parameters and estimation of the vectorial capacity of two subspecies of Triatoma protracta (Uhler) and their laboratory hybrids in Mexico.

Chagas disease is one of the most important vector-borne diseases in Latin America. Studying the biological parameters of each vector species or subspecies contributes to our understanding of their epidemiologic importance. The aim of our study was to compare the biological parameters and start to estimate the vectorial capacity of Triatoma protracta protracta Ryckman, T. p. nahuatlae Ryckman, and their laboratory hybrids. Specifically, we estimated nine biological parameters to increase knowledge about the potential role of triatomine hybrids in the transmission of T. cruzi to reservoir hosts. Nine biological parameters related to the lifecycle, feeding and defecation patterns, number of females, fecundity, and egg eclosion rates in cohorts of T. p. protracta, T. p. nahuatlae, and their hybrids were evaluated and compared. Eight parameters (exception: number of laid eggs) indicated that T. p. nahuatlae was a potentially effective vector of T. cruzi. Our results showed that the hybrid cohorts had better fitness and could potentially have higher vectorial capacity than the parental cohorts. The outstanding characteristics of the hybrids found in our study could lead to an increase in the epidemiologic risks caused by transmission of T. cruzi to humans.

Life cycle of Triatoma huehuetenanguensis Lima-Cordón, Monroy, Stevens, Rodas, Rodas, Dorn, Justi 2019 (Hemiptera: Reduviidae: Triatominae) from Mexico.

Several important biological parameters for estimating the vectorial capacity of a triatomine species were determined on a Mexican population of the recently described Triatoma huehuetenanguensis Lima-Cordón, Monroy, Stevens, Rodas, Rodas, Dorn, Justi (Hemiptera: Reduviidae: Triatominae). The biological parameters were the egg-to-adult development time, number of blood meals required for molting, cumulative mortality, onset time for feeding, feeding and defecation behaviors, and fertility and fecundity of T. huehuetenanguensis. The median egg-to-adult development time was 253 days, with 11 blood meals required for molting. A cumulative mortality rate of 47.41% was observed. The time to begin feeding was between 2.5 and 3.5 min. Feeding times were longer than 15 min. The highest percentages of defecation delays were 1 to 10 min in all nymphal instars and also in males. In contrast, significantly (P < 0.01) more females defecated while eating. At the end of the cycle, the percentage of the obtained females was 54.9%. The mean number of eggs laid per day per female for a month was 2.08. These eggs had an eclosion rate of 98.08%, with an incubation period of 19 days. Our results contribute to estimations of the potential role of T. huehuetenanguensis in the transmission of Trypanosoma cruzi in its distribution area.

Biological Parameters of Two Triatoma rubida Subspecies (Hemiptera: Reduviidae) and Their Laboratory Hybrids.

Chagas disease is one of the most important vector-borne diseases in Latin America, including Mexico. Recently, autochthonous cases have also been detected in the United States of America. It is suspected that two subspecies of Triatoma rubida (Uhler), T. r. sonoriana (Usinger) and T. r. uhleri (Usinger), considered efficient vectors of Trypanosoma cruzi Chagas in Mexico, could interbreed and potentially generate offspring with superior biological characteristics. In this study, the biological parameters of T. r. sonoriana, T. r. uhleri and their laboratory hybrids were evaluated. Hybrids of the two subspecies surpassed both parental subspecies, T. r. sonoriana and T. r. uhleri, in three (numbers of required blood meals to molt [16-20], feeding [10.5-17 min] and defecation times [˂1 min post-feeding]) of nine studied biological parameters. Moreover, the hybrids surpassed T. r. uhleri in two additional parameters, development time (298 d) and mortality (39-42%). Both the hybrid and the two parental cohorts had comparable results in the remaining four (onset of feeding, number of obtained females, number of eggs laid, and eclosion rate) of nine studied parameters. Thus, we conclude that hybrid vigor could result in an increased risk of T. cruzi transmission to humans and animals.

In vitro production of gas methane by tropical grasses / Produccion in vitro de gas metano por gramineas forrajeras tropicales

ABSTRACT Objective. Estimate the production of methane (CH4) by tropical grasses fermented in vitro. Materials and methods. A sample of 20 g dry matter of Cynodon nlemfuensis, Hyparrhenia rufa, Megathyrsus maximus and Digitaria swazilandensis plus 200 ml of culture medium were plated in triplicate flasks sterile stainless steel with CO2 flux, inoculated with 20 ml of ruminal fluid bovine, incubated at 38 °C for 24, 48, 72 and 96 h. Total production of gas, CH4, volatile fatty acids, and pH were evaluated in a completely randomized design with three replicates per treatment and comparison of means with Tukey; the concentration of total and cellulolytic bacteria were analyzed with the Kruskal-Wallis, and the GLM procedure independent data Wilcoxon rank. Results. H. rufa and D. swazilandensis both had the lowest total gas production (p<0.05), while D. swazilandensis had lower production of CH4, increased production of propionic acid (p<0.05) and lower pH 96 hours of incubation (p<0.05). D. swazilandensis showed greater efficiency in energy production due to reduced production of CH4 and increased propionate production. The concentration of total bacteria was similar between treatments (p>0.05), while the concentration of cellulolytic bacteria was lower in C. nlemfuensis y D. swazilandensis when 96 of incubation (p<0.05). Conclusions. The Digitaria swazilandensis, showed favorable conditions to have lower total methane and total gas production.

RESUMEN Objetivo. Estimar la producción de metano (CH4) por gramíneas tropicales fermentadas in vitro. Materiales y métodos. Una muestra de 20 g de materia seca de Cynodon nlemfuensis, Hyparrhenia rufa, Megathyrsus maximus y Digitaria swazilandensis más 200 ml de medio de cultivo se depositaron por triplicado en frascos de acero inoxidable estériles con flujo de CO2, se inocularon con 20 ml de líquido ruminal de bovino e incubaron a 38 °C por 24, 48, 72 y 96 h. Se evaluó producción total de gas, CH4, ácidos grasos volátiles, y pH en un diseño completamente al azar con tres repeticiones por tratamiento y la comparación de medias con Tukey; la concentración de bacterias totales y celulolíticas, se analizaron con la prueba de Kruskal-Wallis, y el procedimiento GLM con datos de rangos independientes de Wilcoxon. Resultados. H. rufa y D. swazilandensis tuvieron la menor producción total de gases (p<0.05), mientras que D. swazilandensis tuvo menor producción de CH4, mayor producción de ácido propónico (p<0.05) y menor pH a las 96 horas de incubación (p<0.05). D. swazilandensis mostró mayor eficiencia en la producción de energía por la menor producción de CH4 y mayor producción de propionato. La concentración de bacterias totales fue similar entre tratamientos (p>0.05), mientras que la concentración de bacterias celulolíticas fue menor en C. nlemfuensis y D. swazilandensis a la hora 96 de incubación (p<0.05). Conclusiones. La Digitaria swazilandensis, mostró condiciones favorables para tener menor producción total de metano y gases totales.

Life history of two abundant populations of Dipetalogaster maximus (Uhler, 1894) (Hemiptera: Reduviidae: Triatominae) in northwestern Mexico.

We focused on the analysis of biological parameters of two different abundant populations of Dipetalogaster maximus (Uhler) (Hemiptera: Reduviidae: Triatominae) from northwestern Mexico. The biological parameters were related to hatching of eggs, life cycle, and meal needed for molting by each instar of two populations from environmentally similar areas (El Fandango and San Dionisio) with a similar number of available hosts as blood meal sources. The D. maximus populations from the two locations were evaluated and compared. No significant differences (P>0.05) were recorded for the average hatching time of the two cohorts. The median egg-to-adult development time and the number of blood meals at each nymphal group were significantly shorter (P<0.05) for the El Fandango cohort. The number of obtained females at the end of the cycles, number of eggs laid, and rate of egg hatching were significantly higher (P<0.05) for the El Fandango cohort. These results contribute to the estimation of abundances of the studied populations of D. maximus in areas where many tourists go for camping and have an increased risk of being bitten and infected by this species.

Chagas disease: Importance of rats as reservoir hosts of Trypanosoma cruzi (Chagas, 1909) in western Mexico.

In Mexico, the role of most species of mammals involved in the transmission cycle of Trypanosoma cruzi Chagas, 1909 is poorly known. It was carried out a study to investigate the importance of rats as reservoir of T. cruzi in western Mexico, an area with important risk of transmission of T. cruzi to human. Thirty-eight human dwellings were searched on two representative towns of western Mexico along twelve months for collection of rats and triatomines. Study rats (Rattus norvegicus) Berkenhout, 1769 and triatomines (Meccus phyllosomus longipennis) (Usinger, 1939) were collected inside and outside human dwellings. Most rats (68.6%, n=312) and triatomines (68.7%, n=217) were collected along months of the hot season. Most rats (59.3%) were collected in peridomiciliary areas. From 312 examined rats, 71 (22.7%) were positive for T. cruzi on examination by Indirect Hemagglutination, which was confirmed by xenodiagnosis. From the 217 examined triatomines, 169 (77.9%) were infected by T. cruzi. The presence of infected rats and triatomines was highly related since on every studied human dwelling where infected triatomines were collected, infected rats were also found. Rats seem to constitute an important domiciliary and peridomiciliary reservoir for T. cruzi, furthering the risk of infection for human beings.

Geographic variation on biological parameters of Meccus picturatus (Usinger), 1939 (Hemiptera: Reduviidae).

The biological parameters related to the life cycles of three populations of Meccus picturatus (Usinger) (Hemiptera: Reduviidae), one of the main vectors of Trypanosoma cruzi Chagas (Trypanosomatida: Trypanosomatidae), were evaluated. A cohort of each of three populations from geographically isolated localities in western Mexico was maintained under similar laboratory conditions, after which all three populations were compared to each other. In each cohort, 50.9 to 72.1% of nymphs completed the cycle. The average time from N1 to adult was approximately 175 days and different for the three studied cohorts. The number of blood meals between molts varied from one to three. The hatching rates as well as the fecundity per female per day were different among the three studied populations. Our results show that the three isolated populations are statistically different in some parameters from each other, which points to the importance of studying the biological characteristics of local populations of triatomines for estimating their capacity of transmitting T. cruzi to reservoir hosts.

Differences on biological attributes of three populations of Meccus pallidipennis Stål (Hemiptera: Reduviidae).

BACKGROUND & OBJECTIVES: Meccus pallidipennis is one of the most epidemiologically important vectors of Trypanosoma cruzi to reservoir hosts in nine states of Mexico. Triatomines occurring in distinct locations normally adapt to local conditions. The aim of this study was to examine the biological attributes of three populations of M. pallidipennis from areas with different environmental characteristics as a factor influencing the triatomine capacity for T. cruzi transmission. METHODS: The values of biological parameters related to the life cycle, the number of blood meals to molt to next instar, fecundity and percentage of females after a biological cycle of three populations of M. pallidipennis were evaluated. A cohort of each of the three studied populations from different geographical areas of Mexico was maintained under similar laboratory conditions and then compared with each other. RESULTS: The life cycle was less than six months in all the studied cohorts, with differences among them. The number of blood meals to molt was lower for the cohort from Izϊcar de Matamoros. Laid eggs per day per female was lower for the cohort from Luvianos. In contrast, no important differences were recorded on the percentage of mortality, egg eclosion rate or percentages of obtained females. INTERPRETATION & CONCLUSION: It was found that an important level of heterogeneity exist between the three studied populations of M. pallidipennis, apparently influenced by the remarkable differences on environmental conditions on the localities where the founders were initially collected, that emphasizes the necessity of studies on local populations of triatomines.

Effect of sugarcane silage on productive parameters of replacement Holstein-Friesian heifers / Efecto del ensilado de caña de azúcar en los parámetros productivos de vaquillas Holstein-Friesian para reemplazo

Objective. Measure the productive parameters of Holstein-Friesian females from weaning to 470 days old, comparing feeding through sugarcane silage (SCS) and corn silage (CS) in a concentrated fodder ratio of 70:30. Materials and methods. Twenty-eight post-weaning calves were used, with an average age of 80±16 days old and an initial weight of 79.5±12.9 kg. The T-Student test was used for the comparison of means, where calves were divided into two groups of 14, one for each treatment. The body weight BW (kg), wither height WH (cm), body condition BC (1 to 5 scale), daily weight gain DWG (kg) and feed conversion FC (kg) was measured every 30 days. Results. Differences were found between treatments, where heifers fed with SCS showed better BC (3.12), FC (7.4) as compared to the CS treatment. No significant differences were found in terms of DWG (0.666 and 0.743 kg/d). Conclusions. Feeding based on sugarcane silage allows for an improved body condition and feed conversion of animals having with this an acceptable and cost-effective production performance in the raising of dairy replacements.

Objetivo. Medir los parámetros productivos de hembras Holstein-Friesian desde el destete hasta los 470 días de edad, comparando la alimentación con ensilado de caña de azúcar (ECA) y ensilado de maíz (EM) en una proporción forraje concentrado de 70:30. Materiales y métodos. Se utilizaron 28 becerras, con una edad promedio de 80±16 días y un peso inicial de 79.5±12.9 kg. Se utilizó la prueba de comparación de medias T- Student, donde se dividieron las becerras en dos grupos de 14, uno para cada tratamiento. Cada 30 días se midió el peso corporal, PC (kg); altura a la cruz, AC (cm); condición corporal, CC (escala de 1 a 5); ganancia diaria de peso, GDP (kg); y conversión alimenticia, CA (kg). Resultados. Se encontraron diferencias entre los tratamientos, donde las vaquillas alimentadas con ECA mostraron mejor CC (3.12), CA (7.4) en comparación con el tratamiento EM. No se encontraron diferencias significativas en cuanto a la GDP (0.666 y 0.743 kg/d). Conclusiones. La alimentación basada en ensilaje de caña de azúcar permite una mejora en la condición corporal y en la conversión alimenticia de los animales teniendo con esto un comportamiento productivo aceptable y económicamente rentable en la crianza de reemplazos lecheros.

Comparative bionomics of four populations of Meccus longipennis (Hemiptera: Reduviidae: Triatominae) under laboratory conditions.

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 biology of three Mexican-American species of Triatominae (Hemiptera: Reduviidae): Triatoma recurva, Triatoma protracta and Triatoma rubida.

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.

The biology of three Mexican-American species of Triatominae (Hemiptera: Reduviidae): Triatoma recurva, Triatoma protracta and Triatoma rubida

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.

Presencia de Meccus longipennis y Triatoma recurva en el estado de Durango, México / Presence of Meccus longipennis and Triatoma recurva in the state of Durango, Mexico

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.

Combining two teaching techniques for young children on Aedes aegypti control: effects on entomological indices in western Mexico.

A study of the effect of educating four- to six-year-old children in mosquito control was recently conducted in a city in the state of Jalisco, western Mexico. Four neighborhood districts were selected. Children attending one kindergarten in each of two experimental districts were taught mosquito control with a video from the American Mosquito Control Association (AMCA), joined to the use of the AMCA Touch Table Technique. The entomological indices monitored in the study decreased significantly (P<0.05) in houses in the experimental districts, apparently because parents acted on the comments and suggestions of the children and eliminated or monitored containers used as oviposition sites by mosquitoes. Based on these results, combining both techniques for teaching children mosquito control is a potentially useful tool for control efforts in Mexico and other places in Latin America.

Distribution and infection of triatomines (Hemiptera: Reduviidae) by Trypanosoma cruzi in the state of Michoacán, Mexico.

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.