Spectral sensitivity of the photonegative reaction of the blood-sucking bug Triatoma infestans (Heteroptera: Reduviidae)

We studied the spectral sensitivity of the visual system of the blood-sucking bug Triatoma infestans, one of the main vectors of Chagas Disease in South America. We quantified the photonegative reaction of this insect in a rectangular arena, half of which was kept dark and the other half illuminated with various intensities of different monochromatic lights (or broadband stimuli for λ>665 nm). As a behavioral parameter of the photonegative response, we measured the time each insect spent in the dark half of the arena. We found that low intensity levels (under 0.06 μW/cm²) of monochromatic lights of 397, 458, 499, and 555 nm evoked a statistically significant (i.e., different from that of control groups) photonegative reaction. Insects were less sensitive to monochromatic lights of 357 nm (UV) and 621 nm (dark orange), and to broadband stimuli in the red part of the spectrum (665–695 nm). These findings indicate that the visual system of T. infestans is sensitive to broader regions of the spectrum than those previously reported.     

Ammonia attracts the haematophagous bug Triatoma infestans : behavioural and neurophysiological data on nymphs

1) Nymphs of the haematophagous bug Triatoma infestans (Heteroptera: Reduviidae) are attracted to volatiles from their own faeces on a servosphere. 2) Biological substrates attractive to triatomines release NH₃: wetted triatomine faecal papers release NH₃ at 256 ppb NH₃ from a 60-g source and stale rabbit urine at 394 ppb from 200 ml. Ammonia released from aqueous NH₃ also attracts bugs at doses of 3 ppb and 17 ppb on the servosphere. 3) Bugs typically show negative anemotaxis in a stimulus-free air-stream on the servosphere. At onset of stimulation with ammonia from either biological substrates or aqueous NH₃ the bugs stop, move their antennae, turn and walk upwind, i.e. odour-mediated anemotaxis. 4) At lower NH₃ doses a latency in attraction is recorded, but this latency disappears when the relative humidity of the stimulus delivery air-stream is dropped from 90 to 35%. 5) Electrophysiological recordings from single olfactory sensilla on antennae of Triatoma nymphs reveal two different types of NH₃-excited receptors, both within grooved-peg sensilla. The responses of one of these receptor cells to NH₃ has been studied in detail and shows that the action potential discharge rate is dose-dependent over the range 2–200 ppb. 6) The amplitudes of electroantennograms recorded from Triatoma nymphs to NH₃ are dose dependent over the range 5–550 ppb. Accepted: 16 January 1997     

Inhibition of xenograft rejection reaction in the bug Triatoma infestans during infection with a protozoan, Trypanosoma cruzi

Autografts and allografts implanted into the body cavity of the bug Triatoma infestans provoked a poor hemocyte encapsulation, while xenografts stimulated significantly stronger reaction. This reaction of xenograft rejection, however, appeared distinctly inhibited in Trypanosoma cruzi-infected bugs. Similar effect was produced in triatomas by injecting them with the fluid from culture of the parasites. The results of these studies suggest that under natural conditions some parasites may avoid destruction and develop within the insect's body cavity following their suppression of host's defense reactions.     

Population dynamics and control of Triatoma infestans

Analysis of field populations of Triatoma infestans (Hemiptera: Reduviidae), after a 3-year study, shows that population growth rate is affected by both density-dependent and density-independent mortality. Although an equilibrium exists, apparently as a consequence of a density dependent-mechanism, population density fluctuates throughout the year because of the effect of monthly mean minimum temperature as a density-independent source of mortality. Simulation studies based on Moran curves shows that high population densities have an approximately constant extinction probability (around 0.20), independently of the season the population starts growing. However, at very low population densities, the extinction probability depends strongly on the season when the population begins to grow. Very low density populations beginning in winter or autumn have the highest extinction probability. The outcome of the simulation studies coincides with results observed in field populations affected by insecticide application at different seasons.     

Chronobiological basis of thermopreference in the haematophagous bug Triatoma infestans

The chronobiological basis of the daily dynamics of thermopreference was tested in adults of Triatoma infestans, continuously registering the preferred temperature of insects released over a temperature gradient for 13 days. We found that the thermopreference in T. infestans is a dynamic process that depends on the time of the day and the post-feeding time. When submitted to a 12:12 h light/darkness cycle (L/D), the preferred temperature reached the highest and the lowest values at the end of the light and dark phases, respectively. This daily rhythm persisted under constant conditions of illumination (D/D and L/L), suggesting the existence of an internal oscillator controlling this behaviour. Statistical analysis revealed that the thermopreference of insects kept under L/D exhibited a ca. 24 h periodicity, while insects kept in D/D and L/L showed free-running periods of tau((D/D))=23.35 and tau((L/L))=27.35 h, respectively. The persistence of a cyclic pattern of thermopreference under constant conditions, and free-running periods, close to, but different from 24 h, demonstrate the existence of an endogenous control of the thermopreference in this species. The biological relevance of these results is discussed in the light of the hypothesis that both the length of time elapsed since feeding and the time of the day modify thermopreference in these bugs. The gradual decline in preferred temperature following feeding may be associated with energy conservation during starvation. The rhythmic modulation of thermopreference may be associated with the daily rhythm of locomotion activity shown by these bugs.     

Isolation and characterization of a cDNA encoding for a lysozyme from the gut of the reduviid bug Triatoma infestans

We have isolated and characterised a Triatoma infestans cDNA encoding a lysozyme. A 174-bp fragment was amplified by PCR using degenerate oligodeoxyribonucleotide primers derived from the known amino acid sequences of lysozyme from other insects. This PCR fragment was used to screen a cDNA gut library of T. infestans. A clone containing the 3'-end of the lysozyme cDNA (219 bp) was isolated and sequenced. RACE was used to amplify the 5'-end of the lysozyme cDNA. After sequencing the complete lysozyme cDNA, the deduced 417 amino acid sequence showed high identity (40-50%) with other chicken-type lysozymes. The amino acid residues responsible for the catalytic activity and the binding of the substrate were essentially conserved. The expression pattern of the lysozyme gene in bugs at different molting and feeding states showed that this gene was upregulated in the digestive tract directly after the molt and after feeding. Additionally, this lysozyme gene was expressed differently in the different regions of the digestive tract, strongly in the cardia and stomach, the anterior regions of the midgut, and only traces of lysozyme mRNA could be detected in the small intestine, the posterior region of the midgut.     

Potential of oil-based formulations of Beauveria bassiana to control Triatoma infestans

The in vitro development of Beauveria bassiana conidia was monitored when immersed in six concentrations of seven non-ionic (MP 6400, MP 600, Renex 60, Renex 95, Span 80, Tween 20 and Tween 80) and three anionic (DOS 75, Hostapaval BVQ 9 and Surfax 220) surfactants and 11 vegetable oils (linseed, soybean, groundnut, rapeseed, thistle, sunflower, olive, sesame, corn, castor, and babassu). The influence of the oils on the settling behavior of Triatoma infestans nymphs and the activity of an oil–water formulation of the fungus against this vector under laboratory and simulated field conditions were also determined. With exception of DOS 75 and Surfax 220 germination of conidia on complete medium was >98% at 24 h after exposure to surfactants up to 10%. Elevated rates of germination (>25%) were observed in 10% corn, thistle and linseed oil 8 days after incubation. Pure oils had a significant repellent effect to T. infestans. Repellency decreased generally at 10% of the oil and some oils showed some attractiveness for nymphs when tested at 1%. Nymphs were highly susceptible to oil–water formulated conidia, even at unfavorable moisture for extra-tegumental development of the fungus on the insect cuticle.     

Visual lateral fixation and tracking in the haematophagous bug Triatoma infestans

Summary Triatoma bugs turn away from a vertical black stripe on the inner surface of a rotating or oscillating drum by keeping it in the lateral visual field at an angle relative to the long axis of the body. The value of depends on the angular velocity w of the drum. Below w=5° s^–1, increases with increasing w, and the stripe can lie to either side of the animal. Occasionally, the bugs switch between these two tracking modes. Above w=5° s^–1, remains nearly constant at about 120° and the stripe lags behind the animal. We call this lateral tracking. At velocities over 5° s^–1 the animals track the leading edge of a wide black stripe in the same manner as they track a narrow stripe. Below 5° s^–1 they walk towards the centre of the stripe (skototaxis). Objects moving towards the insect above the horizon are also fixated at an angle of about 120°. Lateral tracking is mediated mainly by the dorsal part of the visual field, as can be shown by occluding either the dorsal or ventral halves of the eyes. The walking speed of the bugs increases significantly during lateral tracking of an oscillating stripe compared with that during frontal fixation of a stationary one. We therefore interpret lateral tracking as an escape response.     

The response of the blood-sucking bug Triatoma infestans to carbon dioxide and other host odours

Behavioural responses of Triatoma infestans larvae to carbon dioxide and other odours of vertebrate origin were investigated in a locomotion compensator. T. infestans oriented towards airstreams enriched with carbon dioxide exhibiting a threshold response between 300 and 400 p.p.m. above the ambient CO(2) background. The accuracy of the oriented response to carbon dioxide improved with stimulus intensity. Remarkably, insects did not show any change in their sensitivity threshold to carbon dioxide with the starvation time. The attractiveness to carbon dioxide depended on the time of the day, i.e. these nocturnal bugs only oriented towards carbon dioxide-loaded airstreams during the first hours of the scotophase. L-lactic acid did not evoke oriented responses when it was presented as a single stimulus in a wide range of intensities. However, a marked synergism was evident when L-lactic acid was combined with a sub-threshold concentration of carbon dioxide. Under this condition, the threshold response to carbon dioxide decreased to 75-150 p.p.m. above ambient CO(2) background. The isomer D-lactic acid evoked no response, either alone or in combination with carbon dioxide. When insects were stimulated with 1-octen-3-ol a significant positive orientation was found. This response was not modified by the addition of carbon dioxide.     

Differential control of light-dark adaptation in the ocelli and compound eyes of Triatoma infestans

The adaptation to light of compound eyes in insects has been extensively documented and their adaptive role is well understood. Much less attention has been paid, however, to the control of ocelli sensitivity, a study which could help us to understand the functional role of these simple eyes. We analyzed the dynamic changes in the distribution of screening pigments which occur in the ocelli of the haematophagous bug, Triatoma infestans, when the insects are subjected either to light/dark cycles (LD), to constant darkness (DD) or constant light (LL). We then compared these changes with those occurring in the compound eyes of the same individuals and found that, while compound eyes are subject to the control of an endogenous circadian clock, the adaptation of the ocelli is entirely dependent on environmental illumination. In addition, we have observed that environmental temperature is not involved in the control of screening pigments in either ocelli or compound eyes as a direct stimulus, nor as a Zeitgeber. The existence of a differential control in the components of the dual visual system represents an adaptive advantage in the adjustment of visual sensitivity in insects exposed to quick changes in lighting conditions in their natural habitat. We discuss the implications of our findings with regards to the biology of triatomines and with respect to the general understanding the functional role of insect ocelli.     

Aggregation in the haematophagous bug Triatoma infestans: a novel assembling factor

Abstract. The response of Triatoma infestans (Klug) to odours left behind by walking insects was analysed. The insects aggregated on papers marked with putative chemical footprints in the absence of other assembling chemical signals. Contact of the receptor insect with the treated surface was necessary to elicit the response because the signal was not able to attract insects, only to arrest them on marked surfaces. These requirements contrast with other chemical aggregation signals of triatomines that attract the insect and involve olfactory responses. By washing the cuticle of the insects with hexane and impregnating papers with the extract, a response was evoked in test insects similar to that elicited by papers on which insects had walked. The biological role of this novel aggregation factor is discussed in relation to other assembling signals of different origin.     

Dispersive flight by Triatoma infestans under natural climatic conditions in Argentina

Male and female T. infestans were released on two consecutive nights in the salinas of Cordoba Province, Argentina, when air temperatures during the flying period averaged 28.5 degrees C. 136 males (43% of the total released) and 170 females (57%) flew on the first night, and 6 males (18%) and 7 females (27%) on the second. Of these, we recovered 23 males and 14 females within 100 m, and a further female within 200 m, all from the first night's release. The remainder appear to have flown more than 100 m and possibly more than 200 m. In support of this conclusion 4 male and 5 female bugs were recovered in an abandoned brick house 550 m from the release point. Since the proportion of bugs apparently flying more than 200 m is considerably greater than that deduced from previous experiments at lower temperatures (Lehane & Schofield, 1981) it is possible that temperature influences not only the proportion of bugs flying but also the distance flown. Flight appears to be affected by wind speed. On the first night, when there was negligible wind, 43% of male and 57% of female bugs flew. On the second night, with winds gusting at 4-5 m/s, only 18% of the male and 27% of the female bugs flew.     

Brindley's gland exocrine products of Triatoma infestans

olatile exocrine products of the metathoracic Brindley's glands in Triatoma infestans (Hemiptera: Reduviidae) were obtained by dissection and by sampling the air passed over agitated live adults. Isobutyric acid was the main component in the glands, together with isobutyl, isoamyl and amyl alcohols, 2-phenylethanol and other carboxylic acids and esters. Isobutyric acid, isobutyl, isoamyl and amyl alcohols and esters were also found to be emitted into the air, apparently for defence. No volatile products were detected in the metasternal glands.     

Circadian system responses to nocturnal and diurnal hosts in the kissing bug,Triatoma infestans

ABSTRACT

Insects express diverse behavioral rhythms synchronized to environmental cycles. While circadian entrainment to light–dark cycles is ubiquitous in living organisms, synchronization to non-photic cycles may be critical for hematophagous bugs that depend on rhythmic hosts. The purpose was to determine whether Triatoma infestans are capable of synchronizing to the circadian rhythms of potential hosts with temporally distinct activity patterns; and, if so, if this synchronization occurs through masking or entrainment. Precise synchronization with the food source may be critical for the insects’ survival due to the specific predatory or defensive nature of each host. Kissing bugs were housed in a compartment in constant dark, air-flow-connected to another compartment with a nocturnal or a diurnal host; both hosts were synchronized to a light–dark cycle. The activity rhythms of kissing bugs were modulated by the daily activity rhythms of the vertebrates. Effects were a decrease in the endogenous circadian period, independent of the host being nocturnal or diurnal; in some cases relative coordination occurred and in others synchronization was clearly achieved. Moreover, splitting and bimodality arose, phenomena that were also affected by the host presence. The results indicate that T. infestans were able to detect the non-photic cycle of their potential hosts, an ability that surely facilitates feeding and hinders predation risk. Understanding triatomines behavior is of fundamental importance to the design of population control methods.     

Fenitrothion: an alternative insecticide for the control of deltamethrin‐resistant populations of Triatoma infestans in northern Argentina

Deltamethrin‐based campaigns to control Triatoma infestans (Klug) (Hemiptera: Reduviidae) have decreased in success as a result of the development of insecticide resistance. We compared the in vitro effects of the pyrethroid deltamethrin and two doses of the organophosphate fenitrothion, presented on different materials, on T. infestans from La Esperanza, Argentina. Laboratory tests demonstrated a decrease in susceptibility to deltamethrin in the field population [LD₅₀: 30.32 nanograms per insect (ng/i)] compared with the reference population (LD₅₀: 0.13 ng/i), giving a high resistance ratio of 233.42. By contrast, similar susceptibility to fenitrothion was assessed in both the field and reference populations (LD₅₀: 21.65 ng/i and 21.38 ng/i, respectively). The effectiveness of the formulated insecticides varied according to the surfaces to which they were applied. The application of fenitrothion formulations to glass or brick resulted in mortality of 90–100%. The application of fenitrothion formulations to wood or mud caused mortality in the range of 6.7–56.7%. Resistant insects presented low mortality when exposed to the deltamethrin formulation and high mortality when exposed to fenitrothion formulations. Moreover, the insecticides demonstrated residual activity only when applied to glass. The present work demonstrates that fenitrothion is an alternative to pyrethroids for the management of deltamethrin‐resistant insects in La Esperanza. However, this effectiveness is not sustained over time.     

The central projection of cephalic mechanosensory axons in the haematophagous bug Triatoma infestans

The projections of mechanosensory hairs located on the dorsal and lateral head of the adult haematophagous bug Triatoma infestans were analyzed by means of cobalt filling. Axons run into the anterior and posterior tegumentary nerve and project through the brain to the ventral nerve cord. The fibres are small in diameter and run as a fascicle. Some branches run into suboesophageal and prothoracic centres; others run as far as to the mesothoracic ganglion. These sensory projections resemble that of wind-sensitive head hairs of the locust. The functional role of this sensory system in this species is discussed.     

Population dynamics of Triatoma infestans under natural climatic conditions in the Argentine Chaco

Using experimental chicken houses at a site in central Argentina where the bug Triatoma infestans (Klug) is endemic, nine populations of this vector of Chagas disease were monitored during a 34-month period. Bug populations with four chickens as hosts were consistently larger than those with two chickens as hosts. Age structure of the bug population followed a similar pattern irrespective of the initial age structure. Egg to adult mortality was consistently around 98.5% and there was no consistent evidence for density-dependent mortality. There was some evidence for density dependence in fecundity and recruitment rates, but these were heavily constrained by low temperatures during the winter months. Nymphal development rates correlated most strongly with mean minimum temperatures rather than with mean maximum temperatures. We conclude that vector control using insecticides against this species would be most effective at the onset of winter, when recovery of any surviving populations would be inhibited by low temperatures.     

The biting rate of Triatoma infestans in Argentina

1. The daily proportion of fed individuals in a population of the reduviid bug Triatoma infestans (Klug), maintained under natural climatic conditions in experimental chicken houses in central Argentina, was estimated from the proportion of bugs retaining colourless urine in the rectum. 2. From the estimates of feeding frequency throughout a 1 year period, it was shown that temperature has a dominant effect on biting rate, but density-dependent effects became apparent during the warmest months. 3. These and other data on the determinants of blood consumption by T. infestans, were incorporated into a detailed hypothesis of density regulation in this species.     

Mitochondrial 16S DNA variation in populations of Triatoma infestans from Argentina

Abstract.  Variation in the mtDNA 16S ribosomal RNA gene in populations of Triatoma infestans (Klug) was surveyed. DNA sequence comparisons yielded 18 haplotypes among 130 individuals from 16 localities that represent a large proportion of the range of T. infestans in Argentina. The most common genotype in all populations was found in 76.9% of individuals and two other haplotypes were shared among different populations. The remaining 15 haplotypes were present exclusively in one of the populations, suggesting currently low levels of genetic exchange. Analysis of mtDNA 16S sequences uncovered substantial genetic variation among T. infestans populations. Haplotype and nucleotide diversities varied among populations, from 0% to 0.84% and 0% to 0.29%, respectively. It appears that this locus has a low mutation rate. Uncorrected pairwise differences of T. infestans haplotypes ranged from 0% to 1.2%. The molecular phylogeny supported the monophyly of T. infestans haplotypes and clustered two different pairs of haplotypes with a moderate degree of bootstrap support (∼ 60%). Mitochondrial DNA phylogeographic differentiation was not evident, suggesting a recent rapid spread of the species. Analysis of molecular variance showed hierarchical structure in the data. Considerably less variation was found among T. infestans populations from the northwest and northeast regions than among those belonging to the central area. Such a lack of variation may be indicative of one or more past population bottlenecks.