Behavioral evidence of an ectoparasitic interaction between Triatoma pseudomaculata Corrêa e Espínola (Heteroptera: Reduviidae) and Periplaneta americana (L.) (Blattodea: Blattidae)

The present work tested whether Triatoma pseudomaculata Corrêa & Espínola shows behavioral traits indicating that it is capable of feeding on arthropods. We consistently observed nymphs extending their proboscis in the direction of cockroaches and attempting to bite. Insects presented a preference for biting specific cockroach body parts. Our results suggest that arthropod hemolymph represents an alternative source of food that increases the survival of T. pseudomaculata individuals undergoing long starvation.

Misidentification of two Brazilian triatomes, Triatoma arthurneivai and Triatoma wygodzinskyi, revealed by geometric morphometrics.

Triatoma arthurneivai Lent & Martins and Triatoma wygodzinskyi Lent (Hemiptera: Reduviidae) are two Brazilian species found in the sylvatic environment. Several authors may have misidentified T. arthurneivai and consequently published erroneous information. This work reports the use of geometric morphometric analysis on wings in order to differentiate T. arthurneivai and T. wygodzinskyi, and thus to detect possible misidentifications. Triatomines collected from the field in the states of Minas Gerais and São Paulo, and from laboratory colonies, were used. Analyses show a clear differentiation between specimens of T. arthurneivai and T. wygodzinskyi. This indicates that T. arthurneivai populations from São Paulo state were misidentified and should be considered as T. wygodzinskyi. This study also suggests that T. arthurneivai is an endemic species from Serra do Cipó, Minas Gerais state.

Behavioral evidence of an ectoparasitic interaction between Triatoma pseudomaculata Corrêa e Espínola (Heteroptera: Reduviidae) and Periplaneta americana (L.) (Blattodea: Blattidae).

The present work tested whether Triatoma pseudomaculata Corrêa & Espínola shows behavioral traits indicating that it is capable of feeding on arthropods. We consistently observed nymphs extending their proboscis in the direction of cockroaches and attempting to bite. Insects presented a preference for biting specific cockroach body parts. Our results suggest that arthropod hemolymph represents an alternative source of food that increases the survival of T. pseudomaculata individuals undergoing long starvation.

Infestation of peridomestic Attalea phalerata palms by Rhodnius stali, a vector of Trypanosoma cruzi in the Alto Beni, Bolivia.

OBJECTIVES: To determine (i) whether peridomestic Attalea phalerata palms in fragmented human-occupied areas of the Alto Beni, Bolivia, are infested by triatomines; (ii) the specific status of triatomines captured in the area; and (iii) the rate of natural Trypanosoma cruzi infection among those triatomines. METHODS: One hundred and twenty-five live-bait traps were used to sample 47 A. phalerata palms in three Alto Beni localities. Active search for vectors was also performed in 10 chicken coops and three rice storage units. Only Rhodnius specimens were found. As nymphs of closely related Rhodnius species are morphologically undistinguishable, and because of controversy in the literature regarding which Rhodnius species occur in Bolivia, collected insects were identified through molecular taxonomy. Phylogenetic analyses of DNA sequences obtained for a fragment of the mitochondrial cytochrome b gene and for the nuclear ITS-2 ribosomal region were used as molecular markers. Natural infection rates were determined using a pair of primers that PCR-amplify a 330-bp fragment of the parasite's kDNA. RESULTS: Twelve nymphs were captured in five A. phalerata palms (from two of the three localities studied), and an adult was collected from a chicken coop in Iniqua (and morphologically identified as Rhodnius stali). All nymphs (as well as the adult) were molecularly identified as R. stali based on the two molecular markers used. A single nymph was found to be infected with T. cruzi. CONCLUSIONS: Attalea phalerata palms represent an important sylvatic ecotope occupied by R. stali in the Alto Beni region of Bolivia, where there are signs of T. cruzi transmission to humans, despite the preliminary indication of low level of natural infection of the vectors.

New insights on the spread of Triatoma infestans from Bolivia--implications for Chagas disease emergence in the southern cone.

Triatoma infestans, now eliminated from most of South America by control campaigns, has been and still is the main Chagas disease vector due to its ability to colonize rural dwellings. The traditional hypothesis put forth to explain T. infestans adaptation to the synanthropic environment rests on the domestication of wild guinea pigs, one of its natural hosts, by Andean tribes about 5000 BC. Here we present two new hypotheses, based on organized human social activities. The first involves maize production, storage and distribution during the Inca period. Maize granaries could host wild rodent populations that would attract sylvatic T. infestans that were later dispersed during maize distribution. The second hypothesis is associated with the contemporary Urkupiña Virgin festival, near Cochabamba, where thousands of pilgrims gather for rituals in an area that is part of a sylvatic T. infestans focus, thus favoring the contact with the insects and leading to their passive dispersal.

Molecular population genetics and phylogeography of the Chagas disease vector Triatoma infestans in South America.

Knowledge of the genetic variability, population structure, and evolutionary history of Triatoma infestans may be useful for developing rational vector control strategies. A 661-bp fragment of the mitochondrial gene cytochrome oxidase I (COI) was sequenced and analyzed in bugs from Argentina, Uruguay, Peru, and Bolivia, including peridomestic, domestic, Andean, and Chaco sylvatic bugs. A total of 48 polymorphic sites among 37 haplotypes were described. Nucleotide variation fluctuated among samples, with the highest nucleotide diversity observed in seven Argentinean provinces. Within this group, some populations showed patterns of variability compatible with population expansions and/or fine-scale population structure, whereas others suggested population bottlenecks and/or population admixture processes. A maximum parsimony analysis of the haplotypes showed the presence of a Bolivian/Peruvian and an Argentinean/Uruguayan clade. Bolivian sequences were further divided in Chaco sylvatic and Andean domestic and sylvatic. Two different nested clades were found within the Argentinean/Uruguayan cluster. Analysis of molecular variance (AMOVA) and K(ST)* analysis supported a strong population structure in Argentina, where genetic differentiation was correlated with geographic distance. Departures from neutrality expectations and a nested cladistic analysis suggest a recent population expansion of T. infestans in Argentina, followed by restricted gene flow and patterns of isolation by distance. This expansion could have taken place as a two-wave process, as was shown by the phylogenetic analysis and signatures of population admixture in the southern most Argentinean populations.

The association between the geographic distribution of Triatoma pseudomaculata and Triatoma wygodzinskyi (Hemiptera: Reduviidae) with environmental variables recorded by remote sensors.

In this study, predictive models of geographic distribution patterns of Triatoma pseudomaculata (Tps) and T. wygodzinskyi (Twy) were carried out. They were based on biophysical variables estimated from information provided by the satellite remote sensors AVHRR (Advanced Very High Resolution Radiometer) and MODIS (MODerate-resolution Imaging Spectroradiometer). Our goal was to analyze the potential geographic distribution of Tps and Twy and to assess the performance of three predictive models (one for each species and one for both species together) based on temperature, vapour pressure deficit, vegetation and altitude. The geographic distribution analysis shows that all models performed well (>85.7% of overall correct classification of presence and absence point data). The MODIS-based models showed lower correct classifications than the AVHRR-based models. The results strongly suggest that environmental information provided by remote sensors can be successfully used in studies on the geographic distribution of poorly understood Chagas disease vector species.

Microsatellite markers in Triatoma pseudomaculata (Hemiptera, Reduviidae, Triatominae), Chagas' disease vector in Brazil.

Six polymorphic microsatellite loci were isolated and characterized using a microsatellite-enriched genomic library from the Chagas' disease vector Triatoma pseudomaculata. This species is found in Brasil in Caatinga areas and predominantly in peridomestic habitats. All the microsatellites tested on a population of T. pseudomaculata sampled in the Bahia State, Brazil, were polymorphic (2-15 alleles). Markers amplification was also tested on six Triatoma species and some loci successfully amplified in the most phylogenetically related species, in particular Triatoma brasiliensis.

Microsatellite markers from the Chagas disease vector, Rhodnius prolixus (Hemiptera, Reduviidae), and their applicability to Rhodnius species.

Ten microsatellites were isolated and characterized from a partial genomic library of Rhodnius prolixus, the principal Chagas disease vector in Venezuela, Colombia and Central America. These polymorphic molecular markers could be particularly useful in Chagas disease control initiatives. A wider applicability of the primer-pairs isolated was shown, from 6 to 10 loci being amplifiable in five out of the ten Rhodnius species tested, namely R. domesticus, R. nasutus, R. neglectus, R. neivai and R. robustus. Interestingly, all the loci were amplified in the latter. These markers may be of interest to trace the colonization of human dwellings from triatomine sylvatic populations in order to better define epidemiological risk patterns.

Evolutionary relationships based on genetic and phenetic characters between Triatoma maculata, Triatoma pseudomaculata and morphologically related species (Reduviidae: Triatominae).

The maculata group currently comprises two species of Triatominae, Triatoma maculata and Triatoma pseudomaculata, which share morphologic and chromatic characteristics. In order to clarify the systematic status of these two vectors of Trypanosoma cruzi and to infer their evolutionary relationships, we performed an enzymatic, morphometric and cytogenetic comparison of them, also taking into account two sister species not included in the group (T. arthurneivai and T. wygodzinskyi). According to our results, T. maculata and T. pseudomaculata belong to distinct evolutionary lineages. Similarly, T. arthurneivai topotypes from Minas Gerais form an independent isolated group by morphometrics. Our results also support the specific status of the Triatoma population from São Paulo State (formerly referred to T. arthurneivai), and suggest the possibility that it is T. wygodzinskyi. Finally, we suggest that only the arboricolous T. pseudomaculata from northeast Brazil and the rupicolous sister species originated from São Paulo State should be classified together in the same group.

Peridomestic structure, farming activity and triatomine infestation.

The role of peridomestic structure and farming activity on triatomine infestation was studied on two vector species of Chagas disease (Triatoma pseudomaculata and T. brasiliensis) in Bahia State, northeastern Brazil. A randomly selected population issued from 136 farms was divided into four categories according to the householder activity. At regional scale, the dwellings of farmers working on degraded land of irrigated farms are less exposed to T. pseudomaculata infestation. At premises scale, the farmers and casual workers, who have smaller peridomiciles and less cattle, are also less exposed to T. pseudomaculata. The association of T. brosiliensis with the most mobile populations (casual workers and young breeders) suggests a passive transport of this competitive species. Finally, the retired farmers that own large premises and cattle, but have more sedentary behavior, are the most exposed to T. pseudomaculata infestation.

Origin and phylogeography of the Chagas disease main vector Triatoma infestans based on nuclear rDNA sequences and genome size.

For about half of all Chagas disease cases T. infestans has been the responsible vector. Contributing to its genetic knowledge will increase our understanding of the capacity of geographic expansion and domiciliation of triatomines. Populations of all infestans subcomplex species, T. infestans, T. delpontei, T. platensis and T. melanosoma and the so-called T. infestans "dark morph", from many South American countries were studied. A total of 10 and 7 different ITS-2 and ITS-1 haplotypes, respectively, were found. The total intraspecific ITS-2 nucleotide variability detected in T. infestans is the highest hitherto known in triatomines. ITS-1 minisatellites, detected for the first time in triatomines, proved to be homologous and thus become useful markers. Calculations show that ITS-1 evolves 1.12-2.60 times faster than ITS-2. Despite all species analyzed presenting the same n=22 chromosome number, a large variation of the haploid DNA content was found, including a strikingly high DNA content difference between Andean and non-Andean specimens of T. infestans (mean reduction of 30%, with a maximum of up to 40%) and a correlation between presence/absence of minisatellites and larger/smaller genome size. Population genetics analysis of the eight composite haplotypes of T. infestans and net differences corroborate that there are clear differences between western and eastern populations (60%), and little genetic variation among populations (1.3%) and within populations (40%) within these two groups with migration rates larger than one individual per generation corresponding only to pairs of populations one from each of these groups. These values are indicative either of a large enough gene flow to prevent population differentiation by drift within each geographic area or a very recent spread, the latter hypothesis fitting available data better. Phylogenetic trees support a common ancestor for T. infestans and T. platensis, an origin of T. infestans in Bolivian highlands and two different dispersal lines, one throughout Andean regions of Bolivia and Peru and another in non-Andean lowlands of Chile, Paraguay, Argentina, Uruguay and Brazil.

Observations on the domestic ecology of Rhodnius ecuadoriensis (Triatominae).

Rhodnius ecuadoriensis infests peridomiciles and colonises houses in rural southern Ecuador. Six out of 84 dwellings (7%) surveyed in a rural village were infested (78 bugs/infested domicile; 279 bugs were collected in a single dwelling). Precipitin tests revealed R. ecuadoriensis fed on birds (65%), rodents (31%), marsupials (8%), and humans (15%) - mixed bloodmeals detected in 37.5% of individual samples. Trypanosoma cruzi from opossums and rodents may thus be introduced into the domestic cycle. Wasp parasitoidism was detected in 6.5% of 995 R. ecuadoriensis eggs (only in peridomestic habitats). Control strategies should integrate insecticide spraying (indoors and peridomestic), better management of poultry, and housing improvements. A possible inefficacy of Malathion is reported.

Allozyme relationships among ten species of Rhodniini, showing paraphyly of Rhodnius including Psammolestes.

Genetic relationships among 10 species of bugs belonging to the tribe Rhodniini (Hemiptera: Reduviidae), including some important vectors of Chagas disease, were inferred from allozyme analysis of 12 enzyme loci (out of 21 enzyme systems examined), using agarose gel electrophoresis. These species formed two clusters: one comprising Rhodnius brethesi, R. ecuadoriensis, R. pallescens and R. pictipes; the other with Psammolestes tertius, Rhodnius domesticus and the Rhodnius prolixus group comprising R. nasutus, R. neglectus, R. prolixus and R. robustus. The resulting tree was [((R. ecuadoriensis, R. pallescens) R. brethesi) R. pictipes], [R. domesticus (P. tertius [(R. nasutus, R. neglectus) (R. prolixus, R. robustus)])]. Rhodnius nasutus and R. neglectus differed by only one locus, whereas no diagnostic loci were detected between R. prolixus and R. robustus (22 loci were analysed for these four species), despite considerable DNA sequence divergence between species in each of these pairs. Allozymes of the R. prolixus group showed greater similarity with Psammolestes tertius than with other Rhodnius spp., indicating that Rhodnius is paraphyletic and might include Psammolestes.

Observations on the domestic ecology of Rhodnius ecuadoriensis (Triatominae)

Rhodnius ecuadoriensis infests peridomiciles and colonises houses in rural southern Ecuador. Six out of 84 dwellings (7 percent) surveyed in a rural village were infested (78 bugs/infested domicile; 279 bugs were collected in a single dwelling). Precipitin tests revealed R. ecuadoriensis fed on birds (65 percent), rodents (31 percent), marsupials (8 percent), and humans (15 percent) - mixed bloodmeals detected in 37.5 percent of individual samples. Trypanosoma cruzi from opossums and rodents may thus be introduced into the domestic cycle. Wasp parasitoidism was detected in 6.5 percent of 995 R. ecuadoriensis eggs (only in peridomestic habitats). Control strategies should integrate insecticide spraying (indoors and peridomestic), better management of poultry, and housing improvements. A possible inefficacy of Malathion is reported

Nuclear rDNA ITS-2 sequences reveal polyphyly of Panstrongylus species (Hemiptera: Reduviidae: Triatominae), vectors of Trypanosoma cruzi.

Panstrongylus species are widely distributed throughout the Americas, where they act as vectors of Trypanosoma cruzi, agent of Chagas disease. Their intraspecific relationships, taxonomic position and phylogeny in relation to other Triatomini were explored using ribosomal DNA (rDNA) internal transcribed spacer 2 (ITS-2) sequence polymorphisms and maximum parsimony, distance and maximum likelihood analyses of 10 populations representing six species of the genus (P. megistus, P. geniculatus, P. rufotuberculatus, P. lignarius, P. herreri and P. chinai). At the subspecific level, P. megistus appeared more homogeneous than P. rufotuberculatus and P. geniculatus (both with broader distribution). Several dinucleotide microsatellites were detected in the sequences of given species. Many of these microsatellites (GC, TA, GT and AT) showed different number of repeats in different populations and thus, may be very useful for population differentiation and dynamics analyses in future studies. The sequences of P. lignarius (considered sylvatic) and P. herreri (a major disease vector in Peru) were identical, suggesting that these species should be synonymised. Intrageneric analysis showed a clear separation of P. rufotuberculatus, with closest relationships between P. geniculatus and P. chinai, and P. megistus occupying a separate branch. Genetic distances between Panstrongylus species (0.11585-0.22131) were higher than those between Panstrongylus and other Triatomini (16 species from central and North America and South America) (0.08617-0.11039). The distance between P. megistus and P. lignarius/herreri (0.22131) was the largest so far recorded in the tribe. The pronounced differences in length and nucleotide composition suggest a relatively old divergence of Panstrongylus species. P. rufotuberculatus was closer to Mesoamerican Triatoma, Meccus and Dipetalogaster species than to other Panstrongylus. All Panstrongylus clustered with the Mesoamerican clade; P. rufotuberculatus clustered with the phyllosoma complex and T. dimidiata, with D. maxima and T. barberi in a basal position. The rest of Panstrongylus appeared paraphyletically in the tree. This is evidence suggesting polyphyly within the genus Panstrongylus, whose species may be related to the ancestors giving rise to central and North American Triatomini.

Flight and nutritional status of sylvatic Triatoma sordida and Triatoma guasayana.

Nutritional status relating to flight activity was investigated in natural populations of Triatoma sordida and T. guasayana during the dry season in the Bolivian Chaco. The peak flight activity of both species was unimodal and covered the period 61-180 min after sunset. The weight of insects was used as the indicator of nutritional status. Interspecies comparisons employing the same sex and type of capture showed a higher weight for T. guasayana. No significant difference according to weight was observed between flying insects and those collected in natural ecotopes (hollow trees and bromeliads). More than 87% of insects collected from natural ecotopes displayed flight ability under the study conditions, explaining their tendency to invade artificial structures during the dry season.

Natural ecotopes of Triatoma infestans dark morph and other sylvatic triatomines in the Bolivian Chaco.

A survey of natural ecotopes of Triatoma infestans dark morph and other triatomine sylvatic species was performed in an uninhabited area of the Bolivian Chaco. Among the 321 triatomines collected by light trapping, only 4 T. infestans dark morph specimens were identified. Predominant flying species were T. guasayana and T. sordida group 2 (51.7% and 37.1% of capture, respectively). The same species prevailed in terrestrial and epiphytic bromeliads where scarce T. infestans dark morph nymphal instars were also detected. In parrot nests T. delpontei prevailed broadly over other species (90.2% of the capture) and only 4 T. infestans dark morph adults were collected. In contrast, T. infestans dark morph was the predominant species captured in hollow trees (46.0% of the total collected). The abundance of immature forms (88.2% of the collection) shows that hollow trees constitute a favourable ecotope for this species. Of the 421 trees investigated, 33.7% were positive for triatomines. T. infestans dark morph, found inside 15.0% of them, also had higher apparent density than other species (average number of T. infestans in positive trees, 2.0 +/- 1.6 vs 1.3 +/- 0.6 for other species). Light trapping seems to be an efficient method to sample the T. sordida-T. guasayana complex in that it shows a similar distribution to that observed in natural ecotopes; however, this method is ineffective for the assessment of the local abundance of T. infestans dark morph.

Sylvatic population of Triatoma infestans from the Bolivian Chaco: from field collection to characterization.

A sylvatic Triatoma infestans DM (dark morph) population detected in the Bolivian Chaco was characterized and compared with various domestic ones. The degree of differentiation of DM was clearly within the T. infestans intra-specific level. Nevertheless marked chromatic and morphometric differences as well as differences in antennal pattern, chromosome banding and randomly amplified polymorphic DNA support the hypothesis of a distinct population. Continuous exchange of insects between wild and domestic habitats seems unlikely in the Chaco.