Characterization of Feeding Behavior and its Relationship With the Longevity of Wild and Peridomestic Triatoma dimidiata, Latreille 1811 (Hemiptera, Reduviidae) Under Laboratory Conditions.

Triatoma dimidiata (Latreille 1811) is considered the second most important vector of the Trypanosa cruzi etiological agent of Chagas disease in Colombia. It has a life cycle that involves a domiciled, peridomiciled, and wild distribution. The study of feeding behavior and its influence on the survival of sylvatic and peridomestic populations can help identify a possible differential risk in the transmission of Chagas disease to humans, mainly in northwestern and east-central Colombia. We characterize the main parameters of feeding behavior and their influence on the longevity and survival of two rat-fed populations of T. dimidiata from Colombia, one in the north-west (from palms in a tropical dry forest area) and the other in the center-east (peridomiciliated), under controlled environmental conditions. The palm population took considerably longer than the peridomestic population to complete its life cycle under experimental laboratory conditions, being both populations univoltine since they have only one life cycle per year. Statistically significant differences were evidenced using Box-Cox model between the survival rates of T. dimidiata populations when the parameters related to blood intake and behavior were incorporated, in contrast to the survival models in which the origin only was considered as a factor. Our results could be used to generate recommendations to guide prevention strategies in communities near sylvatic and peridomiciliated populations of T. dimidiata.

Environmental Determinants of the Distribution of Chagas Disease Vector Triatoma dimidiata in Colombia.

Triatoma dimidiata (Hemiptera: Reduviidae) is a secondary vector of Trypanosoma cruzi in Colombia and represents an important epidemiological risk mainly in the central and oriental regions of the country where it occupies sylvatic, peridomestic, and intradomestic ecotopes, and because of this complex distribution, its distribution and abundance could be conditioned by environmental factors. In this work, we explored the relationship between T. dimidiata distribution and environmental factors in the northwest, northeast, and central zones of Colombia and developed predictive models of infestation in the country. The associations between the presence ofT. dimidiata and environmental variables were studied using logistic regression models and ecological niche modeling for a sample of villages in Colombia. The analysis was based on the information collected in field about the presence ofT. dimidiata and the environmental data for each village extracted from remote sensing images. The presence of Triatoma dimidiata(Latreille, 1811) was found to be significantly associated with the maximum vegetation index, minimum land surface temperature (LST), and the digital elevation for the statistical model. Temperature seasonality, annual precipitation, and vegetation index were the variables that most influenced the ecological niche model ofT. dimidiata distribution. The logistic regression model showed a good fit and predicted suitable habitats in the Andean and Caribbean regions, which agrees with the known distribution of the species, but predicted suitable habitats in the Pacific and Orinoco regions proposing new areas of research. Improved models to predict suitable habitats forT. dimidiata hold promise for spatial targeting of integrated vector management.

Geographic Distribution and Ecology of Triatoma dimidiata (Hemiptera: Reduviidae) in Colombia.

Triatoma dimidiata Latreille is the second most important vector of Chagas' disease in Colombia and is found in urban and periurban areas. From January 2007 to June 2008, we performed field work in 8 departments, 18 municipalities, and 44 rural villages, covering most of its known distribution and all of its ecological zones in the country. The goal was to determine the geographical distribution, the ecology, and house infestation indices of T. dimidiata over its range and hence the Chagas' disease transmission risk. In Colombia, T. dimidiata occupies a wide variety of ecosystems, from transformed ecosystems in the Andean biome with shrub and xerofitic vegetation to very dense forests in the humid tropical forests in the Sierra Nevada of Santa Marta. According to genetic and ecological criteria, at least two T. dimidiata forms of this species are present: populations from the northwest of the country (Caribbean plains) are restricted to palm tree habitats, and domestic involvement is limited to sporadic visits because of attraction by light; and populations of the east region (Andean mountains) presenting a complex distributional pattern including sylvatic, peridomestic, and domiciliated ecotopes, and occupying a great variety of life zones. The latter population is of epidemiological importance due to the demonstrated migration and genetical flow of individuals among the different habitats. Control, therefore, must take into account its diversity of habitats.

Description and characterization of the melanic morphotype of Rhodnius nasutus Stål, 1859 (Hemiptera: Reduviidae: Triatominae).

INTRODUCTION: For the first time we provide the description of the melanic (dark) morphotype of Rhodnius nasutus and determine the pattern of genetic inheritance for this characteristic. METHODS: Dark morph R. nasutus specimens were crossbred with standard (typically patterned) R. nasutus. RESULTS: We present the first occurrence of the melanic morphotype in the genus Rhodnius. The crossbreeding results demonstrate that the inheritance pattern of this characteristic follows Mendel's simple laws of segregation and an independent assortment of alleles. CONCLUSIONS: Phenotypic variation of R. nasutus reinforces the heterogeneity found in the Triatominae. Descriptions of new species in this subfamily require rigorous validation criteria.

Geometric morphometrics of nine field isolates of Aedes aegypti with different resistance levels to lambda-cyhalothrin and relative fitness of one artificially selected for resistance.

Aedes aegypti, a mosquito closely associated with humans, is the principal vector of dengue virus which currently infects about 400 million people worldwide. Because there is no way to prevent infection, public health policies focus on vector control; but insecticide-resistance threatens them. However, most insecticide-resistant mosquito populations exhibit fitness costs in absence of insecticides, although these costs vary. Research on components of fitness that vary with insecticide-resistance can help to develop policies for effective integrated management and control. We investigated the relationships in wing size, wing shape, and natural resistance levels to lambda-cyhalothrin of nine field isolates. Also we chose one of these isolates to select in lab for resistance to the insecticide. The main life-traits parameters were assessed to investigate the possible fitness cost and its association with wing size and shape. We found that wing shape, more than wing size, was strongly correlated with resistance levels to lambda-cyhalothrin in field isolates, but founder effects of culture in the laboratory seem to change wing shape (and also wing size) more easily than artificial selection for resistance to that insecticide. Moreover, significant fitness costs were observed in response to insecticide-resistance as proved by the diminished fecundity and survival of females in the selected line and the reversion to susceptibility in 20 generations of the non-selected line. As a practical consequence, we think, mosquito control programs could benefit from this knowledge in implementing efficient strategies to prevent the evolution of resistance. In particular, the knowledge of reversion to susceptibility is important because it can help in planning better strategies of insecticide use to keep useful the few insecticide-molecules currently available.

Genetic, cytogenetic and morphological trends in the evolution of the Rhodnius (Triatominae: Rhodniini) trans-Andean group.

The Rhodnius Pacific group is composed of three species: Rhodnius pallescens, R. colombiensis and R. ecuadoriensis, which are considered important vectors of trypanosomes (Trypanosoma cruzi and T. rangeli) infecting humans. This group is considered as a recent trans-Andean lineage derived from the widespread distributed sister taxa R. pictipes during the later uplift of northern Andes mountain range. The widest spread species R. pallescens may be a complex of two divergent lineages with different chromosomal attributes and a particular biogeographical distribution across Central America and Colombia with several southern populations in Colombia occupying the same sylvatic habitat as its sister species R. colombiensis. Although the taxonomy of Rhodnius Pacific group has been well studied, the unresolved phylogenetic and systematic issues are the target of this paper. Here we explore the molecular phylogeography of this species group analyzing two mitochondrial (ND4 and cyt b) and one nuclear (D2 region of ribosomal 28S gene) gene sequences. The molecular analyses suggest an early divergence of the species R. ecuadoriensis and R. colombiensis, followed by a recent expansion of R. pallescens lineages. The phylogenetic relationship between sympatric R. pallescens Colombian lineage and R. colombiensis was further explored using wing morphometry, DNA genome size measurements, and by analyzing chromosomal behavior of hybrids progeny obtained from experimental crosses. Our results suggest that the diversification of the two R. pallescens lineages was mainly influenced by biogeographical events such as (i) the emergence of the Panama Isthmus, while the origin and divergence of R. colombiensis was associated with (ii) the development of particular genetic and chromosomal features that act as isolation mechanisms from its sister species R. pallescens (Colombian lineage). These findings provide new insights into the evolution of the Rhodnius Pacific group and the underlying biological processes that occurred during its divergence.

Eco-geographical differentiation among Colombian populations of the Chagas disease vector Triatoma dimidiata (Hemiptera: Reduviidae).

Triatoma dimidiata is currently the main vector of Chagas disease in Mexico, most Central American countries and several zones of Ecuador and Colombia. Although this species has been the subject of several recent phylogeographic studies, the relationship among different populations within the species remains unclear. To elucidate the population genetic structure of T. dimidiata in Colombia, we analyzed individuals from distinct geographical locations using the cytochrome c oxidase subunit 1 gene and 7 microsatellite loci. A clear genetic differentiation was observed among specimens from three Colombian eco-geographical regions: Inter Andean Valleys, Caribbean Plains and Sierra Nevada de Santa Marta mountain (SNSM). Additionally, evidence of genetic subdivision was found within the Caribbean Plains region as well as moderate gene flow between the populations from the Caribbean Plains and SNSM regions. The genetic differentiation found among Colombian populations correlates, albeit weakly, with an isolation-by-distance model (IBD). The genetic heterogeneity among Colombian populations correlates with the eco-epidemiological and morphological traits observed in this species across regions within the country. Such genetic and epidemiological diversity should be taken into consideration for the development of vector control strategies and entomological surveillance.

Rhodnius barretti, a new species of Triatominae (Hemiptera: Reduviidae) from western Amazonia.

Rhodnius barretti, a new triatomine species, is described based on adult specimens collected in rainforest environments within the Napo ecoregion of western Amazonia (Colombia and Ecuador). R. barretti resembles Rhodnius robustus s.l., but mitochondrial cytochrome b gene sequences reveal that it is a strongly divergent member of the "robustus lineage", i.e., basal to the clade encompassing Rhodnius nasutus, Rhodnius neglectus, Rhodnius prolixus and five members of the R. robustus species complex. Morphometric analyses also reveal consistent divergence from R. robustus s.l., including head and, as previously shown, wing shape and the length ratios of some anatomical structures. R. barretti occurs, often at high densities, in Attalea butyracea and Oenocarpus bataua palms. It is strikingly aggressive and adults may invade houses flying from peridomestic palms. R. barretti must therefore be regarded as a potential Trypanosoma cruzi vector in the Napo ecoregion, where Chagas disease is endemic.

Rhodnius barretti, a new species of Triatominae (Hemiptera: Reduviidae) from western Amazonia

Rhodnius barretti , a new triatomine species, is described based on adult specimens collected in rainforest environments within the Napo ecoregion of western Amazonia (Colombia and Ecuador). R. barretti resembles Rhodnius robustus s.l. , but mitochondrial cytochrome b gene sequences reveal that it is a strongly divergent member of the “robustus lineage”, i.e., basal to the clade encompassing Rhodnius nasutus , Rhodnius neglectus , Rhodnius prolixus and five members of the R. robustus species complex. Morphometric analyses also reveal consistent divergence from R. robustus s.l. , including head and, as previously shown, wing shape and the length ratios of some anatomical structures. R. barretti occurs, often at high densities, in Attalea butyracea and Oenocarpus bataua palms. It is strikingly aggressive and adults may invade houses flying from peridomestic palms. R. barretti must therefore be regarded as a potential Trypanosoma cruzi vector in the Napo ecoregion, where Chagas disease is endemic.

Morphometric and molecular evidence of intraspecific biogeographical differentiation of Rhodnius pallescens (HEMIPTERA: REDUVIIDAE: RHODNIINI) from Colombia and Panama.

Rhodnius pallescens is considered the main vector of Chagas disease in Panama and a relevant secondary vector in northern Colombia. Previous data reported that this species presents cytogenetically heterogeneous populations, which are probably biogeographically segregated. To provide new information on the diversity of R. pallescens, we compared several populations from Colombia and Panama based on the morphometric analyses of wings, mitochondrial cytochrome b (cyt b) gene sequencing, and genomic DNA measurements. Although no differences in DNA amount were detected, significant differences in cyt b sequences as well as wing size and shape were identified among populations. The results obtained in this work indicate R. pallescens comprises two evolutionary lineages with genetic and morphological differences that could be explained by their geographic isolation in distinct ecological zones. These results provide new insight into R. pallescens population diversity and the underlying biological processes that shape its evolution.

Spatial and temporal dynamics of Aedes aegypti larval sites in Bello, Colombia.

Counts of immature stages of the mosquito Aedes aegypti have been used to calculate several entomological indices of dengue vector abundance. Some studies have concluded that these indices can be used as indicators of dengue epidemic risk, while other studies have failed to find a predictive relationship. Ecological niche models have been able to predict distributional patterns in space and time, not only of vectors, but also of the diseases that they transmit. In this study, we used Landsat 7 ETM+ images and two niche-modeling algorithms to estimate the local-landscape ecological niche and the dynamics of Ae. aegypti larval habitats in Bello, Colombia, and to evaluate their potential spatial and temporal distribution. Our models showed low omission error with high confidence levels: about 13.4% of the area presents conditions consistently suitable for breeding across the entire study period (2002-2008). The proportion of neighborhoods predicted to be suitable showed a positive association with dengue case rates, whereas the vector-focused Bretau index had no relationship to case rates. As a consequence, niche models appear to offer a superior option for predictive evaluation of dengue transmission risk and anticipating the potential for outbreaks.

Morphometric and molecular differentiation of a Rhodnius robustus-like form from R. robustus Larousse, 1927 and R. prolixus Stal, 1859 (Hemiptera, Reduviidae).

In Triatominae, "robustus" group constitutes a cluster of species with great haplotypic divergences but high similarities at morphological and nuclear DNA levels. Given these similarities, species identification generates a frequently problematic issue. In northwestern Amazonia, Rhodnius robustus cohabit with an apparently new species, cryptic with R. robustus (Abad-Franch and Monteiro, 2005). In this region (municipality of Puerto Asís, Department of Putumayo, Colombia), we collected insects classified as R. robustus by traditional keys. We compared this sample with specimens of R. robustus from Venezuela, and of R. prolixus from Colombia and Venezuela. The comparisons used landmark-based geometric morphometrics, and analyses of mitochondrial cytochrome b gene and of D2 variable region of the 28S RNA. The shape of the wings from Puerto Asís specimens disclosed clear-cut divergence from the shape of the wings as found for R. prolixus specimens from Venezuela and Colombia, and diverged from the shape of R. robustus from Venezuela. Thus, morphometric analyses suggested that the Puerto Asís collection could represent a new taxon. Using R. pallescens as an outgroup, a tentative phylogenetic tree based on the geometry of the wing showed the Rhodnius from Puerto Asís more similar to the R. prolixus from Colombia than their congeners from Venezuela. In contrast, the molecular classification clustered Colombian R. prolixus and Venezuelan R. robustus with published GenBank sequences, but it gave the insects from Puerto Asís a basal position to the "robustus" group. This outcome suggests that the Puerto Asís haplotype could be the one found by Abad-Franch and Monteiro (2005). Thus, both morphometric and molecular markers used here, although differing in the phylogenetic classification of samples, could differentiate the Puerto Asís sample from the morphologically similar R. prolixus and R. robustus. This could represent a valuable help in the entomological surveillance related to the control of Chagas disease in the South of Colombia and North of Ecuador.

Mapping environmental dimensions of dengue fever transmission risk in the Aburrá Valley, Colombia.

Dengue fever (DF) is endemic in Medellín, the second largest Colombian city, and surrounding municipalities. We used DF case and satellite environmental data to investigate conditions associated with suitable areas for DF occurrence in 2008 in three municipalities (Bello, Medellín and Itagüí). We develop spatially stratified tests of ecological niche models, and found generally good predictive ability, with all model tests yielding results significantly better than random expectations. We concluded that Bello and Medellín present ecological conditions somewhat different from, and more suitable for DF than, those of Itagüí. We suggest that areas predicted by our models as suitable for DF could be considered as at-risk, and could be used to guide campaigns for DF prevention in these municipalities.

Ecology, evolution, and the long-term surveillance of vector-borne Chagas disease: a multi-scale appraisal of the tribe Rhodniini (Triatominae).

Chagas disease incidence has sharply declined over the last decade. Long-term disease control will, however, require extensive, longitudinal surveillance systems capable of detecting (and dealing with) reinvasion-reinfestation of insecticide-treated dwellings by non-domiciliated triatomines. Sound surveillance design calls for reliable data on vector ecology, and these data must cover different spatial scales. We conducted a multi-scale assessment of ecological and evolutionary trends in members of the tribe Rhodniini, including (i) a macroscale analysis of Rhodniini species richness and composition patterns across the Americas, and (ii) a detailed, mesoscale case-study of ecological and behavioural trends in Rhodnius neglectus and R. nasutus. Our macroscale overview provides some comprehensive insights about key mechanisms/processes probably underlying ecological and genetic diversification in the Rhodniini. These insights translate into a series of testable hypotheses about current species distributions and their likely causes. At the landscape scale, we used geometric morphometrics to identify dubious specimens as either R. neglectus or R. nasutus (two near-sibling species), and studied palm tree populations of these two vector taxa in five geographical areas. The data suggest that deforestation and the associated loss of habitat and host diversity might increase the frequency of vector-human contact (and perhaps Trypanosoma cruzi infection rates in vectors). Surveillance in central-northeastern Brazil should prioritise deforested landscapes where large palm trees (e.g., Attalea, Mauritia, Copernicia, Acrocomia or Syagrus) occur near houses. We anticipate that, by helping define the distribution patterns and ecological preferences of each species, multi-scale research will significantly strengthen vector surveillance systems across Latin America.

Morphometric discrimination of females of five species of Anopheles of the subgenus Nyssorhynchus from Southern and Northwest Colombia

The most important vectors of human Plasmodium in the neotropics belong to the subgenus Nyssorhynchus. These species are generally sympatric in terms of their geographical distributions. Some are difficult to identify based solely on examination of adult females using the available morphological keys, in these cases examination of immature stages and male genitalia is required to make correct determinations. However, in epidemiological studies it is necessary to identify the species of adult females which are found near humans, i.e. in studies of malaria transmission or evaluation of control measures. The purpose of the present study was to evaluate the discrimination of adult females of different species of Nyssorhynchus isolated mainly from Southern Colombia (department of Putumayo), using morphometric analysis. Adult females were obtained after rearing larvae collected in natural breeding places and from the progeny of females collected on humans. The morphological characteristics of the immature stages allowed the identification of four species of the subgroup Oswaldoi from Southern Colombia: Anopheles rangeli Gabaldon, Cova Garcia & Lopez, An. oswaldoi (Peryassu), An. benarrochi Gabaldon, Cova Garcia & Lopez and An. triannulatus (Neiva & Pinto). The species An. nuneztovari (Gabaldon) from the Northwest of Colombia was included for comparison. Morphometric analysis allowed differentiation of the females of all species to a confidence level approaching 90 percent using principal components analysis of 10 wing and leg variables, followed by canonical variate analysis of the first four principal components. We conclude that morphometrics may represent a useful taxonomic tool for this group and that its use should be further studied

Geometric morphometric differences between Panstrongylus geniculatus from field and laboratory.

The finding of Panstrongylus geniculatus nymphs inside a house in northeastern Antioquia, Colombia, and the reports related to their increasing presence in homes suggest the need for surveillance methods for monitoring the invasion processes. We analyzed the morphological differences between a wild population and its laboratory descendants, using the techniques of geometric morphometry, with the idea that such differences might parallel those between sylvatic and synanthropic populations. The analyses over five generations showed differences in size but not in shape. Head size and wing size were both reduced from sylvatic to laboratory populations, but the decrease in head size occurred only up to the second generation while the decrease in wing size proceeded up to the fifth generation. In contrast, although a decrease in sexual size dimorphism has been proposed as a marker of colonization in human dwellings, we did not detect any significant loss of dimorphism between sexes of P. geniculatus over the five generations studied. We conclude that size changes may have a physiological origin in response to a change of ecotopes, but more than five generations may be required for the expression of permanent morphological markers of human dwellings colonization.

Geometric morphometric differences between Panstrongylus geniculatus from field and laboratory

The finding of Panstrongylus geniculatus nymphs inside a house in northeastern Antioquia, Colombia, and the reports related to their increasing presence in homes suggest the need for surveillance methods for monitoring the invasion processes. We analyzed the morphological differences between a wild population and its laboratory descendants, using the techniques of geometric morphometry, with the idea that such differences might parallel those between sylvatic and synanthropic populations. The analyses over five generations showed differences in size but not in shape. Head size and wing size were both reduced from sylvatic to laboratory populations, but the decrease in head size occurred only up to the second generation while the decrease in wing size proceeded up to the fifth generation. In contrast, although a decrease in sexual size dimorphism has been proposed as a marker of colonization in human dwellings, we did not detect any significant loss of dimorphism between sexes of P. geniculatus over the five generations studied. We conclude that size changes may have a physiological origin in response to a change of ecotopes, but more than five generations may be required for the expression of permanent morphological markers of human dwellings colonization

Morphometric discrimination of females of five species of Anopheles of the subgenus Nyssorhynchus from Southern and Northwest Colombia.

The most important vectors of human Plasmodium in the neotropics belong to the subgenus Nyssorhynchus. These species are generally sympatric in terms of their geographical distributions. Some are difficult to identify based solely on examination of adult females using the available morphological keys, in these cases examination of immature stages and male genitalia is required to make correct determinations. However, in epidemiological studies it is necessary to identify the species of adult females which are found near humans, i.e. in studies of malaria transmission or evaluation of control measures. The purpose of the present study was to evaluate the discrimination of adult females of different species of Nyssorhynchus isolated mainly from Southern Colombia (department of Putumayo), using morphometric analysis. Adult females were obtained after rearing larvae collected in natural breeding places and from the progeny of females collected on humans. The morphological characteristics of the immature stages allowed the identification of four species of the subgroup Oswaldoi from Southern Colombia: Anopheles rangeli Gabaldon, Cova Garcia & Lopez, An. oswaldoi (Peryassu), An. benarrochi Gabaldon, Cova Garcia & Lopez and An. triannulatus (Neiva & Pinto). The species An. nuneztovari (Gabaldon) from the Northwest of Colombia was included for comparison. Morphometric analysis allowed differentiation of the females of all species to a confidence level approaching 90% using principal components analysis of 10 wing and leg variables, followed by canonical variate analysis of the first four principal components. We conclude that morphometrics may represent a useful taxonomic tool for this group and that its use should be further studied.