Experimentally induced heat-shock tolerance in Panstrongylus megistus (Hemiptera: Reduviidae)

The survival and molting incidence of fifth-instar nymphs of Panstrongylus megistus (Burmeister, 1835), a vector of Chagas' disease, were investigated following sequential heat shocks in which a mild shock (35 or 40 degrees C, 1 h) preceded a more drastic one (40 degrees C, 12 h). The shocks were separated by 8-, 18-, 24-, or 72-h periods at 28 degrees C. The heat-shock tolerance response was more effective when the first shock was given at 40 degrees C. When the period between shocks was 18 h, the tolerance to sequential shocks (in terms of specimen survival) weakened, which suggested a transient control of the process that enables the organism to circumvent the unfavorable effects of severe shock. In terms of molting incidence, the heat-shock tolerance was only demonstrated when the period between the first shock at 40 degrees C for 1 h and the second shock at 40 degrees C for 12 h was > or = 24 h. These results are the first to demonstrate the induction of heat-shock tolerance in a blood-sucking hemipteran.     

Dynamics of thermopreference in the Chagas disease vector Panstrongylus megistus (Hemiptera: Reduviidae)

The thermopreference of resting P. megistus (Burmeister, 1834) adults was studied in a temperature gradient. We also determined the thermopreference for oviposition and ecdysis. Thermopreference of resting individuals depended on the feeding state of the insects. Results demonstrated an initial resting preference of approximately 290 degrees C, which gradually changed to 26-27 degrees C with increasing starvation. A daily variation in thermopreference was also observed with this species. The insects showed a daily maximum preference at the beginning of the dark phase, and a minimum preference during the first half of the light phase. Ecdysis took place all along the gradient, but exhibited a maximum frequency at 30 degrees C. Most eggs were found between 25 and 29 degrees C. Results are discussed in relation to the ecology of P. megistus as compared with other related species, and the existence of mechanisms of behavioral thermoregulation in the group.     

Development cycle of Panstrongylus rufotuberculatus (Hemiptera: Reduviidae) under laboratory conditions

Determinant aspects of the development cycle of Panstrongylus rufotuberculatus (Champion, 1899) were evaluated under laboratory conditions. Fertility, duration of life cycle, mortality rate, feeding time, defecation time, thermal reaction time to food source, and volume of blood ingested were studied. From an initial cohort of fifth instars, nine adult pairs were formed, the females of which oviposited 2,338 eggs over 231 d. The eclosion rate was 84.4%; female and male progeny lived 223.56 (14.41) and 180.78 (14.41) d, respectively. The average total life cycle of immature stages (138.82+/-51.42 d) was distributed as egg, 24.22+/-0.42 d; nymph I, 18.94+/-5.15 d; nymph II, 15.80+/-3.29 d; nymph III, 16.55+/-4.36 d; nymph IV, 25.50+/-7.32 d; and nymph V, 36.71+/-2.75 d. Given these values, 2.6 generations per year are possible. Because of its longevity, rapid response to the presence of a host, large volume of blood ingested, and frequent defecation during feeding, this species constitutes a potentially important vector of Chagas' disease in nonendemic rural areas.     

Cuticular hydrocarbon variability among Triatoma dimidiata (Hemiptera: Reduviidae) populations from Mexico and Guatemala

The geographic variation in the cuticular hydrocarbon pattern among 11 populations of Triatoma dimidiata Latreille (Hemiptera: Reduviidae: Triatominae) from different regions of Mexico and Guatemala, was studied using capillary gas chromatography. T. dimidiata populations were differentiated based on the relative amounts of 71 hydrocarbon components. Insect population classification was mostly in agreement with their geographical vicinity; Mexican populations from the Yucatan peninsula grouped together with those from northern Guatemala, insects from the Mexican Gulf coast states were closely related to those collected from northern Oaxaca, and to a lesser extent, to insects from Chiapas. Insects from southern Oaxaca were clustered together with those from southern Guatemala. All these populations were clearly separated from Guatemalan specimens collected in caves from Alta Verapaz.     

Genetic variability of Triatoma brasiliensis (Hemiptera: Reduviidae) populations

Isoenzymes, random amplified polymorphic DNA (RAPD), and morphometry were used to compare genetic variability within and between three populations of Triatoma brasiliensis Neiva, 1911 from northeast Brazil. The isoenzyme profiles were identical for the three populations, whereas the complex RAPD patterns allowed clear discrimination between the three. Morphometric analysis, using characters of the head capsule, also showed discrimination between the three populations but only in comparisons of males, not females. The results show considerable genetic heterogeneity in T. brasiliensis, with an indication of geographic structuring possibly resulting from a recent series of domestication events.     

Contribution of AT-, GC-, and methylated cytidine-rich DNA to chromatin composition in Malpighian tubule cell nuclei of Panstrongylus megistus (Hemiptera, Reduviidae)

The Malpighian tubule cell nuclei of male Panstrongylus megistus, a vector of Chagas disease, contain one chromocenter, which is composed solely of the Y chromosome. Considering that different chromosomes contribute to the composition of chromocenters in different triatomini species, the aim of this study was to determine the contribution of AT-, GC-, and methylated cytidine-rich DNA in the chromocenter as well as in euchromatin of Malpighian tubule cell nuclei of P. megistus in comparison with published data for Triatoma infestans. Staining with 4′,6-diamidino-2-phenylindole/actinomycin D and chromomycin A₃/distamycin, immunodetection of 5-methylcytidine and AgNOR test were used. The results revealed AT-rich/GC-poor DNA in the male chromocenter, but equally distributed AT and GC DNA sequences in male and female euchromatin, like in T. infestans. Accumulation of argyrophilic proteins encircling the chromocenter did not always correlate with that of GC-rich DNA. Methylated DNA identified by immunodetection was found sparsely distributed in the euchromatin of both sexes and at some points around the chromocenter edge, but it could not be considered responsible for chromatin condensation in the chromocenter, like in T. infestans. However, unlike in T. infestans, no correlation between the chromocenter AT-rich DNA and nucleolus organizing region (NOR) DNA was found in P. megistus.     

Antennal sensilla patterns indicate geographic and ecotopic variability among Triatoma infestans (Hemiptera: Reduviidae) populations

Antennal sensilla patterns were used to explore intraspecific variability among Triatoma infestans (Klug) from domestic habitats in Argentina and Bolivia, and from sylvatic habitats in Bolivia. The sensilla pattern was effective in distinguishing individuals at sexual, ecotopic, and geographic levels, and supported the idea of a lack of gene flow between sylvatic and domestic habitats.     

Intraspecific variability in Triatoma dimidiata (Hemiptera: Reduviidae) populations from Guatemala based on chemical and morphometric analyses

The intraspecific variability of Triatoma dimidiata Latreille, a major vector of Chagas disease, was studied in four departments of Guatemala. Insects were collected from either domestic and sylvatic habitats, and their cuticular hydrocarbon pattern and head morphology were analyzed using ordination and classification techniques. A significant discrimination was obtained both with morphometric and hydrocarbon analyses. Insects from northern departments were easily differentiated from southern conspecifics. Distinctive hydrocarbon pattern and head shape were detected for insects collected from caves in the north central region of the country, posing concern about their taxonomic status.     

Molecular evidence of intraspecific variability in different habitat-related populations of Triatoma dimidiata (Hemiptera: Reduviidae) from Costa Rica

Intraspecific genetic variation among Triatoma dimidiata (Hemiptera: Reduviidae) from seven Costa Rican populations and from different domestic, peridomestic, and sylvatic ecotopes were analyzed. The complete nucleotide sequence of the nuclear ribosomal DNA internal transcribed spacer (ITS-2) and partial sequences of the cytochrome B (Cyt b) gene and the large ribosomal subunit RNA (16S) of mitochondrial DNA (mtDNA) were analyzed and compared. All ITS-2 sequences analyzed were identical and correspond to the haplotype T.dim-H1, the most common haplotype in Central American populations. Sequences of mtDNA revealed a 10.17% of polymorphism in Cyt b and 2.39% in 16S, suggesting that the Cyt b fragment is a useful marker to describe the genetic structure of populations, even at habitat-related level. The analyses of the 18 new combined T. dimidiata haplotypes (Cytb/16S/ITS-2) showed that the two main geographical locations and populations studied are genetically structured showing different haplotype profiling. Only one combined haplotype was shared in the studied areas (Cytb.d/16S.a). Seven haplotypes exclusive for domestic/peridomestic populations, five for sylvatic, and six shared haplotypes for both habitat-related ecotopes are described. Although the relationship between the habitat and the haplotype profiling is less clear, there are different patterns of haplotype distribution in each geographic area between the two habitat-related ecotopes studied (domestic/peridomestic and sylvatic), some of them reflected in the phylogenetic relationships analyzed. The intraspecific variability detected may underlie the known plasticity of T. dimidiata, an important vector for Chagas disease transmission, suggesting that this species must be continuously monitored.     

Wild habitat and related fauna of Panstrongylus lutzi (Reduviidae, Triatominae)

Although Panstrongylus lutzi (Neiva and Pinto) is currently the fifth most frequent species of Triatominae captured in artificial structures in Brazil, its silvatic habitat remains unknown. A survey of its natural ecotopes was performed in an area of Bahia State. P. lutzi nymphal instars and adults were detected in burrows of Dasypodidae. Silvatic habitat of four other endemic triatomine species of the caatinga (Triatoma pseudomaculata Correa and Espinola, Triatoma brasiliensis Neiva, Rhodnius neglectus Lent, and Psammolestes tertius Lent and Jurberg) was also recorded.     

Mitochondrial DNA sequences of triatomines (Hemiptera: Reduviidae): phylogenetic relationships

The phylogenetic relationships among 18 species of Triatominae were inferred based on mitochondrial DNA (mtDNA) sequences. The species of Triatoma included 11 belonging to the infestans complex [T. infestans (Klug), T. guasayana Wygodzinsky & Abalos, T. sordida (St?l), T. platensis Neiva, T. brasiliensis Neiva, T. rubrovaria (Blanchard), T. vitticeps (St?l), T. delpontei Roma?a & Abalos, T. maculata (Erichson), T. patagonica Del Ponte, and T. matogrossensis Leite & Barbosa] and four others of the same genus but of different complexes [T. circummaculata (St?l), T. protracta (Uhler), T. dimidiata (Latreille), and T. mazzottii Usinger]. As possible outgroups we used Mepraia spinolai Mazza, Panstrongylus megistus (Burmeister), and Rhodnius prolixus St?l. We analyzed mtDNA fragments of the 12S and 16S ribosomal RNA genes from each of the 18 species, as well as of the cytochrome oxidase I (COI) gene from nine. The 12S, 16S, and COI gene sequences were analyzed individually and combined. All of the phylogenetic analyses unambiguously supported two clusters: one including T. infestans, T. platensis, and T. delpontei, and the other T. sordida and T. mutagrossensis. Inclusion of T. circummaculata into the infestans complex was confirmed, although this is in disagreement with the morphological classification. On the other hand, our analyses showed that T. dimidiata is closely related to a phylosoma complex species, T. mazzottii. This is consistent with the tentative classification previously made based on morphological characters. The issue of the monophyly of the genus Triatoma remains unresolved.     

Search for a sex pheromone in Triatoma mazzottii (Hemiptera: Reduviidae)

Hexane extracts of volatile substances from sexually mature male and female Triatoma mazzottii Usinger, collected with cold glass trap (-10 degrees C) and evaluated in a wind tunnel, demonstrated the absence of a sex pheromone. The females failed to secrete a liquid from the sixth and seventh abdominal sternite.     

Circadian rhythm of egg hatching in Triatoma infestans (Hemiptera: Reduviidae)

The timing of hatching of Triatoma infestans (Klug) eggs was analyzed by means of time-lapse photography. Under a photoperiod of 12:12 (L:D) and constant darkness, maximum eclosion occurred at the end of the subjective night. Under constant light, the rhythmicity disappeared. Hatching rhythm seems to be maintained by a circadian endogenous oscillator. By hatching close to dawn, when relative humidity is high, insects may avoid excessive water loss.     

Importance and physiological effects of hemolymphagy in triatomines (Hemiptera: Reduviidae)

Triatomines are hematophagous insects and the vectors for Trypanosoma cruzi in the Americas. Although their preferred meal is blood from vertebrate hosts, unfed triatomine nymphs are often seen feeding on different arthropod species. Triatomine saliva has a wide range of activities that aid the hematophagic process. However, nothing is known about its role during hemolymphagy. In the current study, we reproduced hemolymphagy under laboratory conditions and evaluated the influence of hemolymphagy on the survival of Triatoma infestans nymphs. The effects of saliva on the activation of the prophenoloxidase cascade in the invertebrate host and the influence of the saliva on the motility of the bugs and contractions of the dorsal vessels were assessed. Hemolymphagy prolonged the survival rate of T. infestans first instars from 60 to >120 d compared with unfed nymphs. The saliva from T. infestans caused a 50% reduction in the amplitude and frequency of the dorsal vessel contractions of adult Rhodnius prolixus and induced paralysis for >10 min when the saliva was injected into second instars. T. infestans saliva was able to inhibit the activation of the prophenoloxidase cascade from a R. prolixus hemolymph, but had no effect on the phenoloxidase activity after the cascade was activated. The paralyzing molecule in the saliva was <5 kDa and probably had no proteic or lipidic characteristics. These results suggest that triatomine saliva may play an important role during hemolymphagy by inducing paralysis and suppressing immune responses in the invertebrate host. The importance of hemolymphagy for the survival of nymphs in periods when vertebrate blood is scarce is also discussed.