Marcelo G. Lorenzo

Chemical communication in chagas disease vectors. source, identity, and potential function of volatiles released by the metasternal and Brindley's glands of Triatoma infestans adults

Compounds from the metasternal and Brindleys glands of the blood-sucking bug, Triatoma infestans, were identified by solid phase microextraction (SPME) and gas chromatography-mass spectrometry. Volatile compounds released by adult bugs during copulation or after mechanical disturbance were also characterized. Six compounds were identified and found consistently in all samples from metasternal glands. The most abundant were 3-pentanone, 2-methylbutanol, 3-pentanol, and an unidentified compound. The metasternal gland blends did not differ qualitatively between sexes. Compounds found in Brindleys glands were short chain acids, alcohols, esters, and a ketone with no qualitative differences between sexes. Isobutyric acid was the main component of this blend, and two new confirmed compounds were described as products of these glands: 2-butanone and 2-methylbutyric acid. 3-Pentanone was collected from the headspace over 33% of the copulating pairs of T. infestans. Volatiles found in the headspace of disturbed T. infestans adults included short-chain fatty acids, alcohols, esters, and ketones, with no qualitative differences between sexes. Both types of glands apparently discharge their contents after disturbance. However, most of the volatiles released by bugs after disturbance came from Brindleys glands. The locomotor activity of fourth instars increased significantly after stimulation with the odors emitted by disturbed adults, as compared with larvae stimulated by the odor of undisturbed adults or by clean air. We also studied the directional behavioral response of fifth instars to the disturbance scent in a locomotion compensator. Larvae exposed to volatiles released by disturbed adults walked away from the direction of the odor. The results suggest that this blend or part of it functions as an alarm pheromone for T. infestans. We suggest that the metasternal glands of this species are involved both in the sexual and the alarm contexts, and that the Brindleys glands probably have both alarm and defensive roles.

Metasternal Gland Volatiles and Sexual Communication in the Triatomine Bug, Rhodnius prolixus

Twelve compounds produced by the metasternal glands (MGs) of the triatomine bug Rhodnius prolixus were identified by solid phase microextraction (SPME) combined with coupled gas chromatography-mass spectrometry (GC-MS) using achiral and chiral columns. All substances were ketones or alcohols, and the same compound profile was found in the secretions produced by either sex. The most abundant compounds were 2-methyl-3-buten-2-ol, (2S)-pentanol, (3E)-2-methyl-3-penten-2-ol, and (2R/2S)-4-methyl-3-penten-2-ol. Emission of these compounds was detected more frequently from females than males, and females released them more frequently during the early hours of the scotophase, the period when sexual activity in this species is at its peak. These compounds were also detected in the headspace above mating pairs. Finally, the occlusion of the MG orifices of male or female bugs with paraffin resulted in a significant decrease in copulation frequency compared to sham-operated insects. Together, these data suggest that the MG secretions of R. prolixus may be involved in sexual communication.

Biology, diversity and strategies for the monitoring and control of triatomines: Chagas disease vectors

Despite the relevant achievements in the control of the main Chagas disease vectors Triatoma infestans and Rhodnius prolixus, several factors still promote the risk of infection. The disease is a real threat to the poor rural regions of several countries in Latin America. The current situation in Brazil requires renewed attention due to its high diversity of triatomine species and to the rapid and drastic environmental changes that are occurring. Using the biology, behaviour and diversity of triatomines as a basis for new strategies for monitoring and controlling the vectorial transmission are discussed here. The importance of ongoing long-term monitoring activities for house infestations by T. infestans, Triatoma brasiliensis, Panstrongylus megistus, Triatoma rubrovaria and R. prolixus is also stressed, as well as understanding the invasion by sylvatic species. Moreover, the insecticide resistance is analysed. Strong efforts to sustain and improve surveillance procedures are crucial, especially when the vectorial transmission is considered interrupted in many endemic areas.

The effect of relative humidity on the behaviour and development of Triatoma brasiliensis

Abstract The preference for relative humidity (RH) and suitability of different levels of this environmental parameter were investigated in the haematophagous bug Triatoma brasiliensis Neiva, 1911 (Hemiptera, Reduviidae). The hygropreference of T. brasiliensis was studied using a RH gradient and the effect of different RHs on the egg hatching, nymph mortality and moulting success was also analysed. The results show that egg hatching in first‐instar nymphs of T. brasiliensis was lower at extreme RHs and, particularly, it was lowest at 9.3% RH. The survival of starved nymphs was not affected by RH, but the percentage of engorged nymphs and the ecdysis success of these nymphs once fed was diminished strongly by high humidity. Fourth‐instar nymphs preferred to stay at the lowest RH during the first 5 days after feeding and during ecdysis. This preference changed markedly during starvation. Fifteen days after ecdysis, the bugs moved towards intermediate humidities, and 30 days after ecdysis they even preferred the most humid sectors of the gradient. Females preferred to lay eggs in dry environments, suggesting that they may not have a particular hygropreference for oviposition, but that they simply lay their eggs at the RHs where they prefer to stay.

Do Haematophagous Bugs Assess Skin Surface Temperature to Detect Blood Vessels

Background It is known that some blood-sucking insects have the ability to reach vessels under the host skin with their mouthparts to feed blood from inside them. However, the process by which they locate these vessels remains largely unknown. Less than 5% of the skin is occupied by blood vessels and thus, it is not likely that insects rely on a “random search strategy”, since it would increase the probability of being killed by their hosts. Indeed, heterogeneities along the skin surface might offer exploitable information for guiding insects bites. Methodology/Principal Findings We tested whether the bug Rhodnius prolixus can evaluate temperature discontinuities along the body surface in order to locate vessels before piercing the host skin. When placed over a rabbit ear, the bugs first bites were mostly directed towards the main vessels. When insects were confronted to artificial linear heat sources presenting a temperature gradient against the background, most bites were directly addressed to the warmer linear source, notwithstanding the temperature of both, the source and the background. Finally, tests performed using uni- and bilaterally antennectomized insects revealed that the bilateral integration of thermal inputs from both antennae is necessary for precisely directing bites. Conclusions/Significance R. prolixus may be able to exploit the temperature differences observed over the skin surface to locate blood vessles. Bugs bite the warmest targets regardless of the target/background temperatures, suggesting that they do not bite choosing a preferred temperature, but select temperature discontinuities along the skin. This strategy seems to be an efficient one for finding blood vessels within a wide temperature range, allowing finding them on different hosts, as well as on different areas of the host body. Our study also adds new insight about the use of antennal thermal inputs by blood sucking bugs.

Behavioral and Electrophysiological Responses of Triatoma brasiliensis Males to Volatiles Produced in the Metasternal Glands of Females

In many insects, mate finding is mediated by volatile sex pheromones, but evidence for this phenomenon in triatomines (Heteroptera: Reduviidae) is still fragmentary. Recently, it was shown that metasternal glands (MGs) are involved in producing signals related to the sexual communication of Triatoma infestans and Rhodnius prolixus. Based on this, we tested whether MG volatiles could be involved in the sexual communication of Triatoma brasiliensis. Odor-mediated orientation responses were studied by using a T-tube olfactometer. These tests showed that males exhibit positive anemotaxis when confronted with adult odor-laden air currents. Moreover, females that had their metasternal glands occluded did not elicit significant orientation by males. Compounds produced by the MGs of T. brasiliensis females were identified by means of SPME, GC-FID, and GC-MS, with achiral and chiral columns. All substances identified were ketones and alcohols, and similar compound profiles were found in the secretions produced by both sexes. The most abundant compounds identified were 3-pentanone, followed by (4R)-methyl-1-heptanol, 3-pentanol, and (2S)-methyl-1-butanol. In addition, GC-EAD recordings showed that the antennae of males responded to several of the main components of female MG secretions. Our results showed that compounds produced by the MGs of T. brasiliensis females are involved in the sexual communication of this species.

Behavioural biology of Chagas disease vectors

Many arthropod species have adopted vertebrate blood as their main food source. Blood is rich in nutrients and, except for the presence of parasites, sterile. However, this food source is not freely available, nor is obtaining it devoid of risk. It circulates inside vessels hidden underneath the skin of mobile hosts that are able to defend themselves and even predate the insects that try to feed on them. Thus, the haematophagous lifestyle is associated with major morphological, physiological and behavioural adaptations that have accumulated throughout the evolutionary history of the various lineages of blood-sucking arthropods. These adaptations have significant consequences for the evolution of parasites as well as for the epidemiology of vector-transmitted diseases. In this review article, we analyse various aspects of the behaviour of triatomine bugs to illustrate how each behavioural trait represents a particular adaptation to their close association with their hosts, which may easily turn into predators. Our aim is to offer to the reader an up-to-date integrative perspective on the behaviour of Chagas disease vectors and to propose new research avenues to encourage both young and experienced colleagues to explore this aspect of triatomine biology.

Copulation and mate guarding behavior in Triatoma brasiliensis (Hemiptera: Reduviidae).

ABSTRACT The hematophagous bug Triatoma brasiliensis (Neiva) is an important vector of Chagas disease in Brazil. We describe the mating behavior of this species in a comparative manner and analyze the effect of time elapsed since the first imaginal bloodmeal on motivation of males to mate, on female mate-rejections, and on overall mating frequency. We also evaluate whether females of this species copulate with several males in short time intervals, and we examine whether the duration of copulation may vary in the presence of other males compared with what is observed with isolated pairs. Age affected the motivation of males to perform mating attempts, whereas nutritional status affected the receptivity of females. Results showed that 70% of the females tested accepted copulating with three males over a period of 90 min, whereas the remaining 30% mated with at least two males. Mating duration in T. brasiliensis was prolonged in the presence of other males compared with observations of isolated pairs, and males of this species performed long postcopulatory associations only in the presence of other males, apparently as a mate guarding response to the presence of potential competitors.

Aggregation behaviour in Panstrongylus megistus and Triatoma infestans: inter and intraspecific responses☆

We tested the aggregation response to inter and intraspecific chemical signals in Panstrongylus megistus and Triatoma infestans. As previously described for T. infestans, larvae of P. megistus significantly aggregated on papers impregnated with their own excrement and on papers marked with cuticular substances deposited on surfaces on which these insects had walked. T. infestans bugs also aggregated on papers impregnated by faeces or by cuticular substances of P. megistus, and P. megistus aggregated on papers contaminated by faeces or by cuticular substances of T. infestans. The response of P. megistus to its cuticular substances was significantly stronger than that to its faeces. The non-specificity of the two signals is discussed in the context of the ecological relationship between both species.

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

Abstract 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 ≈29°C, which gradually changed to 26–27°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°C. Most eggs were found between 25 and 29°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.