Paz María Salazar-Schettino

Phylogeography and Genetic Variation of Triatoma dimidiata, the Main Chagas Disease Vector in Central America, and Its Position within the Genus Triatoma

Background Among Chagas disease triatomine vectors, the largest genus, Triatoma, includes species of high public health interest. Triatoma dimidiata, the main vector throughout Central America and up to Ecuador, presents extensive phenotypic, genotypic, and behavioral diversity in sylvatic, peridomestic and domestic habitats, and non-domiciliated populations acting as reinfestation sources. DNA sequence analyses, phylogenetic reconstruction methods, and genetic variation approaches are combined to investigate the haplotype profiling, genetic polymorphism, phylogeography, and evolutionary trends of T. dimidiata and its closest relatives within Triatoma. This is the largest interpopulational analysis performed on a triatomine species so far. Methodology and Findings Triatomines from Mexico, Guatemala, Honduras, Nicaragua, Panama, Cuba, Colombia, Ecuador, and Brazil were used. Triatoma dimidiata populations follow different evolutionary divergences in which geographical isolation appears to have had an important influence. A southern Mexican–northern Guatemalan ancestral form gave rise to two main clades. One clade remained confined to the Yucatan peninsula and northern parts of Chiapas State, Guatemala, and Honduras, with extant descendants deserving specific status. Within the second clade, extant subspecies diversity was shaped by adaptive radiation derived from Guatemalan ancestral populations. Central American populations correspond to subspecies T. d. dimidiata. A southern spread into Panama and Colombia gave the T. d. capitata forms, and a northwestern spread rising from Guatemala into Mexico gave the T. d. maculipennis forms. Triatoma hegneri appears as a subspecific insular form. Conclusions The comparison with very numerous Triatoma species allows us to reach highly supported conclusions not only about T. dimidiata, but also on different, important Triatoma species groupings and their evolution. The very large intraspecific genetic variability found in T. dimidiata sensu lato has never been detected in a triatomine species before. The distinction between the five different taxa furnishes a new frame for future analyses of the different vector transmission capacities and epidemiological characteristics of Chagas disease. Results indicate that T. dimidiata will offer problems for control, although dwelling insecticide spraying might be successful against introduced populations in Ecuador.

Chromosomal variation and genome size support existence of cryptic species of Triatoma dimidiata with different epidemiological importance as Chagas disease vectors

The wide geographical distribution of Triatoma dimidiata, one of the three major vectors of Chagas disease, ranges from Mexico to northern Peru. Since this species occupies a great diversity of artificial and natural ecotopes, its eradication is extremely difficult. In order to assist control efforts, we used chromosome analyses and DNA amount as taxonomic markers to study genetic variability in populations of T. dimidiata from Mexico, Guatemala, El Salvador and Colombia. We differentiated three groups or cytotypes defined by characteristic chromosome C‐banding patterns and genome size measured by flow cytometry. The three cytotypes are restricted to different geographic locations. Cytotype 1 occurs in Mexico (excluding Yucatán), Guatemala (excluding Petén), El Salvador and Colombia. Cytotype 2 occurs in Yucatán and cytotype 3 occurs in Petén. Cytotype 1, commonly associated with domestic and peridomestic environments but also inhabiting sylvatic ecotopes, is the most widespread and with major epidemiological significance. In contrast, the Yucatán cytotype inhabits wild ecotopes but increasingly enters houses, while the Petén cytotype appears exclusively sylvatic. We suggest that these cytotypes represent cryptic species of T. dimidiata with different epidemiological relevance as Chagas disease vectors. Poor ability to colonize human dwellings, together with their restricted geographic distribution, indicate that the Yucatán and Petén putative species probably have much less epidemiological significance than cytotype 1. Thus, the genetic markers we describe are powerful tools to differentiate cryptic species in T. dimidiata with different epidemiological significance, contributing to planning the most effective control measures.

Unusual prevalence of the Giardia intestinalis A-II subtype amongst isolates from humans and domestic animals in Mexico

Defined by Filice in 1952 as Giardia duodenalis (syn. Giardia lamblia) on the basis of morphological characteristics, the species includes organisms that are ubiquitous in a variety of mammalian hosts. However, this morphological similarity masks genetic and biotypic differences that are sufficiently large for G. intestinalis now to be considered a species complex. Seven major sublineages (A–G) representing widely distributed genotypes, some with discernable host range differences, have been defined by phylogenetic analyses (Monis et al., 1999; Thompson et al., 2000).

Metric Variation Among Geographic Populations of the Chagas Vector Triatoma dimidiata (Hemiptera: Reduviidae: Triatominae) and Related Species

Abstract Eight Triatoma dimidiata populations from different geographic regions were compared along with related species using traditional morphometry of head characters. A method for removing allometric change was used for the intraspecific comparisons, and scaling for the isometric change of size was used for the interspecific comparisons. The intraspecific comparison showed significant head shape differences between T. dimidiata populations correlating with geography, showing a separation between the northern, intermediate, and southern collections (more evident in females), and supporting the idea that this species includes several evolutionarily divergent populations. The positioning of one sylvatic group from Guatemala did not correlate with geography, because it was more closely related to a distinct population of Colombia. This sylvatic group was found in caves, while the Colombian specimens, although collected in houses, may have migrated from nearby caves. Evolutionary and/or ecological influences could be responsible for the head shape similarities between these two groups: a common ancestral origin of both populations or a morphological convergence caused by similar environmental pressures. The interspecific comparisons included four other regional species of the same genus, three of them belonging to the phyllosoma complex (T. pallidipennis, T. mexicana, and T. ryckmani, the latter provisionally) and the fourth one classified in the protracta complex (T. nitida). Both complexes were readily separated by their head dimensions, even after size adjustment, and our data support inclusion of T. dimidiata within the phyllosoma complex.

Antennal Phenotype of Triatoma dimidiata Populations and Its Relationship with Species of phyllosoma and protracta Complexes

Abstract Triatoma dimidiata (Latreille 1811) Reduviidae Triatominae is the main vector of Chagas disease in several countries of Latin America. As for other vector species, the characterization of T. dimidiata subpopulations within particular geographical regions or occupying different habitats could help in better planning of vector control actions. A first objective in this study was to evaluate the antennal phenotype as a phenetic marker to characterize populations of T. dimidiata collected in different geographic areas and domestic and sylvatic habitats. A second objective was to evaluate the phenetic relationships of T. dimidiata with other species of the phyllosoma complex: longipennis, pallidipennis, and phyllosoma. The antennal sensilla of T. dimidiata specimens collected in Mexico, Central America, and Colombia were analyzed and compared with the antennal sensilla of T. longipennis, T. pallidipennis, and T. phyllosoma. T. barberi was used as an outgroup in the analysis. For each specimen, the ventral side of the three distal segments of the antennae was drawn, identifying and counting four types of sensilla. In T. dimidiata, univariate and multivariate analysis showed differences between sexes, among populations collected in different habitats within the same region, and among populations collected in different geographic regions. Two types of antennal sensilla showed a latitudinal variation. Domestic specimens showed intermediate characteristics of the antennal phenotype, between sylvatic cave- and sylvatic forest-collected specimens. The antennal phenotypes show high similarities among T. pallidipennis, T. phyllosoma, and T. longipennis, with a better differentiation of T. pallidipennis. T. dimidiata is separated from the other members of the complex by a similar distance to T. barberi, of the protracta complex.

Phylogeographic Pattern and Extensive Mitochondrial DNA Divergence Disclose a Species Complex within the Chagas Disease Vector Triatoma dimidiata

Background Triatoma dimidiata is among the main vectors of Chagas disease in Latin America. However, and despite important advances, there is no consensus about the taxonomic status of phenotypically divergent T. dimidiata populations, which in most recent papers are regarded as subspecies. Methodology and Findings A total of 126 cyt b sequences (621 bp long) were produced for specimens from across the species range. Forty-seven selected specimens representing the main cyt b clades observed (after a preliminary phylogenetic analysis) were also sequenced for an ND4 fragment (554 bp long) and concatenated with their respective cyt b sequences to produce a combined data set totalling 1175 bp/individual. Bayesian and Maximum-Likelihood phylogenetic analyses of both data sets (cyt b, and cyt b+ND4) disclosed four strongly divergent (all pairwise Kimura 2-parameter distances >0.08), monophyletic groups: Group I occurs from Southern Mexico through Central America into Colombia, with Ecuadorian specimens resembling Nicaraguan material; Group II includes samples from Western-Southwestern Mexico; Group III comprises specimens from the Yucatán peninsula; and Group IV consists of sylvatic samples from Belize. The closely-related, yet formally recognized species T. hegneri from the island of Cozumel falls within the divergence range of the T. dimidiata populations studied. Conclusions We propose that Groups I–IV, as well as T. hegneri, should be regarded as separate species. In the Petén of Guatemala, representatives of Groups I, II, and III occur in sympatry; the absence of haplotypes with intermediate genetic distances, as shown by multimodal mismatch distribution plots, clearly indicates that reproductive barriers actively promote within-group cohesion. Some sylvatic specimens from Belize belong to a different species – likely the basal lineage of the T. dimidiata complex, originated ∼8.25 Mya. The evidence presented here strongly supports the proposition that T. dimidiata is a complex of five cryptic species (Groups I–IV plus T. hegneri) that play different roles as vectors of Chagas disease in the region.

FIRST CASE OF NATURAL INFECTION IN PIGS. REVIEW OF TRYPANOSOMA CRUZI RESERVOIRS IN MEXICO

An epidemiological research project was performed in the State of Morelos including collection of samples for blood smears and culture, serological tests, and xenodiagnoses from a total of 76 domestic and peridomestic mammals. Two strains of Trypanosoma cruzi were isolated by haemocultures; one from a pig (Sus scrofa), the first case of natural infection reported in Mexico, and the other from a dog (Canis familiaris). This study summarizes current information in Mexico concerning confirmed reservoirs of T. cruzi.

Enteroparasitosis en poblaciones indígenas y mestizas de la Sierra de Nayarit, México

We compared the prevalences of enteroparasitoses from 420 individuals, 306 of them wereindigenous and 114 were mestizo persons, without gender differentiation, and of all ages from theNayarit mountain range. Six field trips were made to the region during one year to collect the stoolsamples, some individuals provided only one sample and other even five. Stool samples weresuspended 1:5 with 10% formalin for their transportation to Mexico City, where they were processedby direct parasitoscopic analysis with lugol solution We found

Chagas disease as a cause of symptomatic chronic myocardopathy in Mexican children.

We report the first case series of children in Mexico living with symptomatic Chagas disease causing chronic myocardopathy. The findings suggest that children with Chagas disease may develop symptomatic chronic myocardopathy earlier than previously recognized. Our findings emphasize the importance of longitudinal cardiologic follow-up of all children identified with acute Chagas disease. In a cohort of 826 children from the state of Queretaro in Mexico, 11 were identified with positive serology (ELISA and IFI) for Chagas and were tested for electrocardiogram alterations and symptoms and signs. Four children had ECG alterations with 3 of these reporting signs and symptoms associated with the chronic phase of Chagas disease (27%; 95% CI: 6%–61%). The most common chronic symptom was chest pain, with one child also reporting dyspnea and tachycardia.

Occurrence of hybrids and laboratory evidence of fertility among three species of the Phyllosoma complex (Hemiptera: Reduviidae) in Mexico

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.