Morphometric analysis of Triatoma dimidiata populations (Reduviidae:Triatominae) from Mexico and Northern Guatemala

Triatoma dimidiata is one of the major vectors of Chagas disease in Latin America. Its range includes Mexico, all countries of Central America, Colombia, and Ecuador. In light of recent genetic analysis suggesting that the possible origin of this species is the Yucatan peninsula, we have analyzed populations from the state of Yucatan, San Luis Potosi, and Veracruz in Mexico, and a population from the southern region of the Yucatan peninsula located in Northern Guatemala, the region of El Peten. Classical morphometry including principal component, discriminant, sexual dimorphism, and wing asymmetry was analyzed. San Luis Potosi and Veracruz populations were indistinguishable while clearly separate from Yucatan and Peten populations. Despite important genetic differences, Yucatan and Peten populations were highly similar. Yucatan specimens were the smallest in size, while females were larger than males in all populations. Only head characters were necessary to distinguish population level differences, although wing fluctuating asymmetry was present in all populations. These results are discussed in light of recent findings suggesting genetic polymorphism in most populations of Triatoma dimidiata south of Chiapas to Ecuador.

A survey on Triatoma dimidiata in an urban area of the province of Heredia, Costa Rica

Triatoma dimidiata has been found in several cities and towns of those countries where the insect is a domestic or peridomestic pest. In Central America, urban infestations occur in the capitals of at least five countries. During 2001 and 2002 a survey was carried out in the county of San Rafael, Heredia province, located 15 km northwest of San José, capital of Costa Rica, in order to determine the degree of infestation by T. dimidiata in an entire city block. Six peridomestic colonies of the insect were detected in the backyards of eight households. The ecotopes occupied by the insects consisted of store rooms with old objects, wood piles or firewood, and chicken coops. A total of 1917 insects were found in the six foci, during two sampling periods, and a mean infection rate by Trypanosoma cruzi of 28.4% was found in 1718 insects examined. The largest colony found in one of the households yielded 872 insects that were thriving mainly at the expenses of two dogs. Opossums and adult insects were common visitors of the houses and it became evident that this marsupial is closely related to the peridomestic cycle of the Chagas disease agent. Lack of colonization of the insect inside the human dwellings is explained by the type of construction and good sanitary conditions of the houses, in contrast to the situation in most peridomiciliary areas. Stomach blood samples from the insects showed that the main hosts were, in order of decreasing frequency: rodents, dogs, fowl, humans, opossums, and cats. The fact that no indication of infection with Chagas disease could be detected in the human occupants of the infested houses, vis a vis the high infection rate in dogs, is discussed.

Maintenance and breeding of Thrichomys (Trouessart, 1880) (Rodentia: Echimyidae) in captivity

South American histricognath rodents Thrichomys apereoides laurentius and Thrichomys pachyurus are natural hosts of Trypanosoma cruzi, agent of Chagas disease. We established breeding colonies of these species to serve as experimental models in various parasitological studies. Both species of Thrichomys have all the requirements necessary to become excellent laboratory models: they can be easily maintained in the standard laboratory conditions and breed throughout the year and they do not have any special dietary demands and can be fed by standard food pellets designed for laboratory mice. Both species produce precocious offspring that have their eyes and ears open, teeth erupted, fur well developed, and can eat solid food in the first week of life. T. a. laurentius has larger litter sizes and lower body masses at birth and weaning than T. pachyurus. Moreover, females of T. a. laurentius reach puberty earlier and with lower body mass than T. pachyurus.

Morfologia comparada dos ovos e ninfas de Triatoma vandae Carcavallo, Jurberg, Rocha, Galvão, Noireau & Lent, 2002 e Triatoma williami Galvão, Souza & Lima, 1965 (Hemiptera, Reduviidae, Triatominae)

Eggs and all nymphs of these species were studied employing light microscopy (LM) and scanning electron microscopy (SEM). The major differences observed by LM in the eggs were related to the presence and the distribution of pores on the surface of their chorion. Morphological differences among three nymphal stages (1st, 3rd, and 5th) development of each species were observed. The differential characteristics are chromatic and in the shape of connexival spots. The ultrastructure of the ventral region of the head and the IX, X, and XI abdominal segments (anal tube) of the both species were described demonstrating morphological differences that can be used for diagnosis of the species.

Patterns of antenal sensilla of Panstrongylus megistus from three Brazilian states

The objective of the present study was to analyze and describe the phenotype of the antennal sensilla of Panstrongylus megistus, one of the epidemiologically most important species of triatomines in Brazil. Specimens from the Brazilian states of Goiás (GO), Minas Gerais (MG), and Rio Grande do Sul (RS) were compared, based on studies of four types of sensilla on three antennal segments: thick-walled trichoid (TK), thin-walled trichoid (TH), bristles (BR), and basiconica (BA). Discriminant analysis allowed the separation of the RS specimens from those of GO and MG. Multivariate discriminant analysis demonstrated that the sensilla of males differed from those of females, the variables with greatest weight being the BA of all three segments and the TK of flagellum 1. The basiconica sensilla were significantly more abundant in females, on all three segments. Antennal sensilla patterns also demonstrated significant differences among P. megistus specimens.

Genetic diversity of Triatoma infestans (Hemiptera: Reduviidae) in Chuquisaca, Bolivia based on the mitochondrial cytochrome b gene

Partial cytochrome b DNA sequences for 62 Triatoma infestans were analyzed to determine the degree of genetic variation present in populations of this insect in the northwest region of Chuquisaca, Bolivia. A total of seven haplotypes were detected in the localities sampled. The phylogenetic relationship and population genetic structure of the haplotypes found in this region, indicate that there is greater variation in this relatively small region of Bolivia than what has been previously reported by studies using the same gene fragment, for more distant geographic areas of this country. In addition, a comparison of rural and peri-urban localities, indicate that there is no difference in the genetic variation of T. infestans between these two environments.

Antileishmanial and trypanocidal activity of Brazilian Cerrado plants

The side effects and the emerging resistance to the available drugs against leishmaniasis and trypanosomiasis led to the urgent need for new therapeutic agents against these diseases. Thirty one extracts of thirteen medicinal plants from the Brazilian Cerrado were therefore evaluated in vitro for their antiprotozoal activity against promastigotes of Leishmania donovani, and amastigotes of Trypanosoma cruzi. Among the selected plants, Casearia sylvestris var. lingua was the most active against both L. donovani and T. cruzi. Fifteen extracts were active against promastigotes of L. donovani with concentrations inhibiting 50% of parasite growth (IC50) between 0.1-10 µg/ml, particularly those of Annona crassiflora (Annonaceae), Himatanthus obovatus (Apocynaceae), Guarea kunthiana (Meliaceae), Cupania vernalis (Sapindaceae), and Serjania lethalis (Sapindaceae). With regard to amastigotes of T. cruzi, extracts of A. crassiflora, Duguetia furfuracea (Annonaceae), and C. sylvestris var. lingua were active with IC50 values between 0.3-10 µg/ml. Bioassay fractionations of the more active extracts are under progress to identify the active antiparasite compounds.

Analysis of antenal sensilla patterns of Rhodnius prolixus from Colombia and Venezuela

Antennal sensilla patterns were used to analyze population variation of domestic Rhodnius prolixus from six departments and states representing three biogeographical regions of Colombia and Venezuela. Discriminant analysis of the patterns of mechanoreceptors and of three types of chemoreceptors on the pedicel and flagellar segments showed clear differentiation between R. prolixus populations east and west of the Andean Cordillera. The distribution of thick and thin-walled trichoids on the second flagellar segment also showed correlation with latitude, but this was not seen in the patterns of other sensilla. The results of the sensilla patterns appear to be reflecting biogeographic features or population isolation rather than characters associated with different habitats and lend support to the idea that domestic R. prolixus originated in the eastern region of the Andes.

Morphobiological aspects of Rhodnius brethesi Matta, 1919 (Hemiptera:Reduviidae) from the Upper and Middle Negro River, Amazon region of Brazil: I - scanning electron microscopy

The occurrence of autochthonous cases of Chagas disease in the Amazon region of Brazil over recent decades has motivated an intensification of studies in this area. Different species of triatomines have been identified, and ten of these have be proven to be carriers of the parasite Trypanosoma cruzi or " cruzi-like " parasites. Studies conducted in the municipalities of Santa Isabel do Rio Negro and Barcelos, located on the Upper and Middle of the Negro River, microregion of Negro River, state of Amazonas have confirmed not only that Rhodnius brethesi is present in the palm tree Leopoldinia piassaba, but also that this insect was recognized by palm fiber collectors. A morphological study of eyes, inter-ocular and inter-ocellar regions, antennae, buccula, labrum, rostrum, stridulatory sulcus and feet, including the apex of the tibia, spongy fossette and ctenidium was conducted by scanning electron microscopy. The buccula and the stridulatory sulcus presented notable differences in specimens of different genera and also of different species. These data make it possible to suggest that the details presented in these structures can be included as diagnostic characteristics to be used in new dichotomous keys, thereby contributing towards studies of taxonomy and systematics and furnishing backing for comparative analysis of specimens collected from different localities.

CC-chemokine receptors: a potential therapeutic target for Trypanosoma cruzi-elicited myocarditis

The comprehension of the pathogenesis of Trypanosoma cruzi-elicited myocarditis is crucial to delineate new therapeutic strategies aiming to ameliorate the inflammation that leads to heart dysfunction, without hampering parasite control. The augmented expression of CCL5/RANTES and CCL3/MIP-1alpha, and their receptor CCR5, in the heart of T. cruzi-infected mice suggests a role for CC-chemokines and their receptors in the pathogenesis of T. cruzi-elicited myocarditis. Herein, we discuss our recent results using a CC-chemokine receptor inhibitor (Met-RANTES), showing the participation of CC-chemokines in T. cruzi infection and unraveling CC-chemokine receptors as an attractive therapeutic target for further evaluation in Chagas disease.

Genetic structure of Triatoma venosa (Hemiptera: Reduviidae): molecular and morphometric evidence

Triatoma venosa presents a restricted geographical distribution in America and is considered as a secondary vector of Chagas disease in Colombia and Ecuador. A total of 120 adult insects were collected in domestic and peridomestic habitats in an endemic area of the department of Boyacá, Colombia, in order to determine their genetic structure through morphometric and molecular techniques. The head and wings of each specimen were used for the analyses of size, shape, and sexual dimorphism. A significant sexual dimorphism was found, although no differences in size among the studied groups were detected. Differences were found in the analyzed structures except for male heads. DNA was extracted from the legs in order to carry out the internal transcriber space-2 (ITS-2) amplification and the randon amplified polymorphic DNA (RAPD) analyses. Length polymorphisms were not detected in the ITS-2. Fst and Nm values were estimated (0.047 and 3.4, respectively). The high genetic flow found among the insects captured in the domicile and peridomiciliary environment does not permit a genetic differentiation, thus establishing the peridomicile as an important place for epidemiological surveillance.

Distribution and ecological aspects of Rhodnius pallescens in Costa Rica and Nicaragua and their epidemiological implications

In light of the Central American Initiative for the control of Chagas disease, efforts were made on the part of Costa Rican and Nicaraguan teams, working separately, to determine the present status of Rhodnius pallescens in areas close to the common border of the two countries, where the insect has appeared within the last few years. The opportunity was also used to establish whether R. prolixus, a vector present in some areas of Nicaragua, has been introduced in recent years into Costa Rica with Nicaraguan immigrants. It became evident that wild adults of R. pallescens are common visitors to houses in different towns of a wide area characterized as a humid, warm lowland, on both sides of the frontier. Up to the present, this bug has been able to colonize a small proportion of human dwellings only on the Nicaraguan side. There was strong evidence that the visitation of the adult bug to houses is related to the attraction of this species to electric lights. There were no indications of the presence of R. prolixus either in Nicaragua or in Costa Rica in this area of the Caribbean basin. Triatoma dimidiata, a widespread domestic species in both countries, was totally absent in the explored areas of Costa Rica but occasionally occurs on the Nicaraguan side. Serological surveys in children of both areas showed that transmission of Chagas disease takes place in a rather small degree in Costa Rica and more commonly in Nicaragua, indicating that R. pallescens could be a potential threat as a vector in this particular region.

Arylfurans as potential Trypanosoma cruzi trypanothione reductase inhibitors

The natural lignans veraguensin and grandisin have been reported to be active against Trypanosoma cruzi bloodstream forms. Aiming at the total synthesis of these and related compounds, we prepared three 2-arylfurans and eight 2,5-diarylfurans. They were evaluated for their potential as T. cruzi trypanothione reductase (TR) inhibitors as well against the parasite's intracellular (amastigote) and bloodstream (trypomastigote) forms. Compound 12 was the most effective against TR with an IC50 of 48.5 µM while 7 and 14 were active against amastigotes, inhibiting the parasite development by 60% at 20 µg/ml (59 and 90 µM, respectively). On the other hand, none of the compounds was significantly active against the parasite bloodstream forms even at 250 µg/ml (0.6-1.5 mM).

The effect of Bulgarian propolis against Trypanosoma cruzi and during its interaction with host cells

Propolis has shown activity against pathogenic microorganisms that cause diseases in humans and animals. The ethanol (Et-Blg) and acetone (Ket-Blg) extracts from a Bulgarian propolis, with known chemical compositions, presented similar activity against tissue culture-derived amastigotes. The treatment of Trypanosoma cruzi-infected skeletal muscle cells with Et-Blg led to a decrease of infection and of the intracellular proliferation of amastigotes, while damage to the host cell was observed only at concentration 12.5 times higher than those affecting the parasite. Ultrastructural analysis of the effect of both extracts in epimastigotes revealed that the main targets were the mitochondrion and reservosomes. Et-Blg also affected the mitochondrion-kinetoplast complex in trypomastigotes, offering a potential target for chemotherapeutic agents.

Molecular characterization of human Trypanosoma cruzi isolates from endemic areas in Panama

The present work provides information on Trypanosoma cruzi genotype circulating in endemic areas of Chagas disease in Panama. A total of 26 crude stocks of T. cruzi, isolated from the blood of persons with different clinical profiles of Chagas disease were collected and crio-conserved until used. Most of the stocks had been characterized by means of isoenzyme electrophoresis on cellulose acetate membranes. The clinical profiles of infected persons included 9 (34.6%) asymptomatic and 17 acute (65.4%) including 5 (19.2%) fatal cases, 2 under 5 years old and 3 adults. A multiplex-PCR assay based on the amplification of the non-transcribed spacer of the mini-exon gene was performed. All stocks of T. cruzi included in the study were found to correspond to Tc I group. This result supports the predominance of T. cruzi-I in the transmission cycles affecting the human population in the Republic of Panama.