Barometric pressure decrease induces density-dependent changes in foraging behaviour in a parasitoid fly.

Barometric pressure is an environmental factor involved in the modulation of a variety of activities in insects. Generally, a drop in barometric pressure precedes the arrival of weather conditions that can affect insect activities and life expectancy. We simulated different scenarios of pressure drop in a modified hermetic chamber and studied their influence on the host-seeking behaviour of the larvae of the robber fly Mallophora ruficauda using air stationary olfactometers. In addition, we studied whether larval density modulates orientation to the host under the same scenarios of pressure drop. We found that motivation to search for hosts is affected by the same slope of pressure drop in both low- and high-density larvae. However, larval density modulates the onset of the responses to pressure decrease, as low-density larvae stop searching for hosts more quickly than high-density larvae. This result reflects an avoidance strategy according to which low-density larvae would have a reduced host range and higher risk of mortality and fewer chances to find a suitable host under adverse pressure conditions. Low-density larvae, known to prefer healthy hosts, do not search for parasitized hosts under normal pressure conditions nor under a range of pressure drops, strongly suggesting that host selectivity is not modulated by barometric pressure. This study paves the way to a better understanding of the changes in crucial insect behaviours induced by weather conditions, and provides more knowledge about the risk factors likely to affect insect survival in the context of foraging ecology.

Mosquitoes do not Like Bitter.

Chemical repellents play a crucial role in personal protection, serving as essential elements in reducing the transmission of vector-borne diseases. A biorational perspective that extends beyond the olfactory system as the classical target may be a promising direction to move. The taste system provides reliable information regarding food quality, helping animals to discriminate between nutritious and potentially harmful food sources, often associated with a bitter taste. Understanding how bitter compounds affect feeding in blood-sucking insects could unveil novel molecules with the potential to reduce biting and feeding. Here, we investigated the impact of two naturally occurring bitter compounds, caffeine and quinine, on the feeding decisions in female Aedes aegypti mosquitoes at two distinctive phases: (1) when the mosquito explores the biting substrate using external taste sensors and (2) when the mosquito takes a sip of food and tastes it using internal taste receptors. We assessed the aversiveness of bitter compounds through both an artificial feeding condition (artificial feeder test) and a real host (arm-in-cage test). Our findings revealed different sensitivities in the external and internal sensory pathways responsible for detecting bitter taste in Ae. aegypti. Internal detectors exhibited responsiveness to lower doses compared to the external sensors. Quinine exerted a more pronounced negative impact on biting and feeding activity than caffeine. The implications of our findings are discussed in the context of mosquito food recognition and the potential practical implications for personal protection.

Microorganismal Cues Involved in Host-Location in Asilidae Parasitoids.

Parasitoids are organisms that kill their host before completing their development. Typical parasitoids belong to Hymenoptera, whose females search for the hosts. But some atypical Diptera parasitoids also have searching larvae that must orientate toward, encounter, and accept hosts, through cues with different levels of detectability. In this work, the chemical cues involved in the detection of the host by parasitoid larvae of the genus Mallophora are shown with a behavioral approach. Through olfactometry assays, we show that two species of Mallophora orient to different host species and that chemical cues are produced by microorganisms. We also show that treating potential hosts with antibiotics reduces attractiveness on M. ruficauda but not to M. bigoti suggesting that endosymbiotic bacteria responsible for the host cues production should be located in different parts of the host. In fact, we were able to show that M. bigoti is attracted to frass from the most common host. Additionally, we evaluated host orientation under a context of interspecific competence and found that both parasitoid species orient to Cyclocephaala signaticollis showing that host competition could occur in the field. Our work shows how microorganisms mediate orientation to hosts but differences in their activity or location in the host result in differences in the attractiveness of different cues. We show for the first time that M. bigoti behaves similar to M. ruficauda extending and reinforcing that all Mallophora species have adopted a parasitoid lifestyle.

Species-Specificity in Thermopreference and CO2-Gated Heat-Seeking in Culex Mosquitoes.

Combining thermopreference (Tp) and CO2-gated heat-seeking assays, we studied the thermal preferendum and response to thermal cues in three Culex mosquito species exhibiting differences in native habitat and host preference (e.g., biting cold and/or warm-blooded animals). Results show that these species differ in both Tp and heat-seeking behavior. In particular, we found that Culex territans, which feed primarily on cold-blood hosts, did not respond to heat during heat-seeking assays, regardless of the CO2 concentration, but exhibited an intermediate Tp during resting. In contrast, Cx. quinquefasciatus, which feeds on warm blooded hosts, sought the coolest locations on a thermal gradient and responded only moderately to thermal stimuli when paired with CO2 at higher concentrations. The third species, Cx. tarsalis, which has been shown to feed on a wide range of hosts, responded to heat when paired with high CO2 levels and exhibited a high Tp. This study provides the first insights into the role of heat and CO2 in the host seeking behavior of three disease vectors in the Culex genus and highlights differences in preferred resting temperatures.

How Did Seal Lice Turn into the Only Truly Marine Insects?

Insects are the most evolutionarily and ecologically successful group of living animals, being present in almost all possible mainland habitats; however, they are virtually absent in the ocean, which constitutes more than 99% of the Earth's biosphere. Only a few insect species can be found in the sea but they remain at the surface, in salt marshes, estuaries, or shallow waters. Remarkably, a group of 13 species manages to endure long immersion periods in the open sea, as well as deep dives, i.e., seal lice. Sucking lice (Phthiraptera: Anoplura) are ectoparasites of mammals, living while attached to the hosts' skin, into their fur, or among their hairs. Among them, the family Echinophthiriidae is peculiar because it infests amphibious hosts, such as pinnipeds and otters, who make deep dives and spend from weeks to months in the open sea. During the evolutionary transition of pinnipeds from land to the ocean, echinophthiriid lice had to manage the gradual change to an amphibian lifestyle along with their hosts, some of which may spend more than 80% of the time submerged and performing extreme dives, some beyond 2000 m under the surface. These obligate and permanent ectoparasites have adapted to cope with hypoxia, high salinity, low temperature, and, in particular, conditions of huge hydrostatic pressures. We will discuss some of these adaptations allowing seal lice to cope with their hosts' amphibious habits and how they can help us understand why insects are so rare in the ocean.

Under pressure: the extraordinary survival of seal lice in the deep sea.

Lice from pinnipeds - sea lions, seals and walruses - are the only insects capable of surviving marine dives. Throughout their evolutionary history, they have adapted to tolerate hypoxia, high salinity, low temperature and, in particular, to tolerate conditions of high hydrostatic pressure. To understand the limits of the capacity of lice to survive during host deep dives, we conducted a series of controlled experiments in the laboratory. We collected lice from elephant seals and submitted the different life stages to high pressure conditions. Lice were first exposed to one of four hydrostatic pressures: 30, 80, 150 or 200 kg cm-2 They were then exposed a second time to higher or lower hydrostatic pressure conditions to test for the impact of the first experience, which could either be deleterious or trigger physiological adaption, allowing them a better tolerance to high pressure. We found that lice from elephant seals can tolerate hydrostatic pressures higher than 200 kg cm-2 (close to 200 atm), which is equivalent to 2000 m depth. Adults exhibited lower recovery times than nymphs after immersion at high hydrostatic pressure. Our findings show that lice have developed unique adaptations to endure extreme marine conditions. We discuss these extreme performances in relation to the morphological characteristics and physiological responses to diving in these insects.

Thermosensation and the TRPV channel in Rhodnius prolixus.

The thermal sense of triatomine bugs, vectors of Chagas disease, is unique among insects. Not only do these bugs exhibit the highest sensitivity to heat known in any animal up to date, but they can also perceive the infrared radiation emitted by the body of their warm-blooded hosts. The sensory basis of this capacity has just started to be unravelled. To shed additional light on our understanding of thermosensation, we initiated an analysis of the genetic basis of the thermal sense in Rhodnius prolixus. We tested the hypothesis that a TRPV (transient receptor potential vanilloid) channel receptor is involved in the evaluation of heat in this species. Two different approaches were adopted. Initially, we analysed the expression of a TRPV candidate for this function, i.e., RproIav, in different tissues. Subsequently, we tested the effects of capsaicin and capsazepine, two molecules known to interact with mammal TRPV1, using three different behavioural protocols for evaluating thermal responses: (1) proboscis extension response (PER), (2) thermopreference in a temperature gradient and (3) spatial learning in an operant conditioning context. Bioinformatic analyses confirmed that the characteristic features typical of the TRPV channel subfamily are found in the RproIav protein sequence. Molecular analysis showed that RproIav is expressed in R. prolixus, not only in the antennae, but also in other body structures bearing sensory organs. Behavioural experiments consistently revealed that capsaicin treated insects are less responsive to heat stimuli and prefer lower temperatures than non-treated insects, and that they fail to orient in space. Conversely, capsazepine induces the opposite behaviours. The latter data suggest that triatomine thermoreception is based on the activation of a TRP channel, with a similar mechanism to that described for mammal TRPV1. The expression of RproIav in diverse sensory structures suggests that this receptor channel is potentially involved in bug thermoreception. This constitutes solid evidence that thermosensation could be based on the activation of TRP receptors that are expressed in different tissues in R. prolixus. Whether RproIav channel is a potential target for the compounds tested and whether it mediates the observed effects on behaviour still deserves to be confirmed by further research.

Barometric pressure influences host-orientation behavior in the larva of a dipteran ectoparasitoid.

Rain and temperature have been awarded as the most important weather factors that influence insect behavior. Barometric pressure studies have been relegated to a secondary place mainly because most studies deal with adult insects where temperature and water availability are the main environmental factors that influence behavior. We studied the influence of barometric pressure on the host orientation behavior in Mallophora ruficauda, an ectoparasitoid with an active host-seeking larval stage. Our results show that a steeper decrease in barometric pressure than expected by regular variation reduced orientation to host chemical cues. This study is the first to show a correlation between changes in the barometric pressure and the seeking behavior of parasitoid soil-dwelling larvae. Our results show that in this kind of insects, ambient factors other than temperature, water availability and light, can influence and have a profound impact on the process of parasitism. We discuss the influence of this behavior on a task so important for parasitoids as host location, and highlight the importance of including such information in parasitoid foraging ecology and climatic change studies.

Incidence of Non-Immunological Defenses of Soil White Grubs on Parasitism Success of Mallophora ruficauda Larva (Diptera: Asilidae).

White grubs are larvae of Coleoptera of the family Scarabaeidae. They are known because of their intensive feeding habits on crop roots. Mallophora ruficauda (Diptera: Asilidae) is a dipteran parasitoid whose larva is a natural enemy for white grubs. This species is a solitary ectoparasitoid, where both female and larva realize different steps in the host location process. Female place its eggs in high grasslands and then, the larva finds and parasitizes the host in the ground. There are nine potential hosts in the area of action of this parasitoid; however a high preference for Cyclocephala signaticollis has been observed (87% of field parasitism). It is known that many insects have developed defensive and immunological mechanisms when attacked by a parasitoid, which can be behavioral, physiological, chemical or genetic. The objectives of this work were to investigate what kind of defense and non-immunological associated mechanisms the white grubs have against this parasitoid and to understand why M. ruficauda have such a high preference for masked chafer grubs or Cyclocephala species. In particular, for each white grub species, we asked: (1) If there is a differential behavioral reaction when a parasitoid attack is simulated; (2) If body attributes of white grubs species have influence on defense behavior, and particularly for the masked chafer C. signaticollis; and (3) Why this species is the most selected by M. ruficauda. It was found that behavioral defenses of white grubs would explain the parasitism pattern of M. ruficauda larvae and its preference for C. signaticollis.

Orientation mechanisms and sensory organs involved in host location in a dipteran parasitoid larva.

The robber fly Mallophora ruficauda is one of the principal pests of apiculture in the Pampas region of Argentina. Larvae are solitary ectoparasitoids of third-instar scarab beetle larvae. Females of M. ruficauda do not lay eggs on or near the hosts, but on tall grasses. After hatching, larvae are dispersed by the wind and drop to the ground, where they dig and search for potential hosts. It is known that second-instar larvae of M. ruficauda exhibit active host-searching behaviour towards their preferred hosts, i.e., third-instar larvae of Cyclocephala signaticollis. Although host-location seems to be mediated by chemical cues, the mechanism of orientation and the sensory organs involved in host location remain unknown. We carried out behavioural experiments in the laboratory to address these questions. We also tested whether the orientation behaviour is exclusively based on the use of chemical cues. We found that larvae of M. ruficauda detect the chemicals with chemosensilla on the maxillary palps. Only one maxillary palp suffices for orientation, but their bilateral ablation abolishes orientation. Besides, an hexane extract of the host body was as attractive as a live host. Our results support that M. ruficauda larvae find their hosts underground by means of chemoklinotaxis.

Insights to host discrimination and host acceptance behaviour in a parasitoid (Diptera: Asilidae): implications for fitness.

The robber fly Mallophora ruficauda is one of the principal pests of apiculture in the Pampas region of Argentina. As adults they prey on honey bees and other insects, while as larvae they are solitary ectoparasitoids of third instar scarab beetle larvae. Females of M. ruficauda lay eggs away from the host in tall grasses. After being dispersed by the wind, larvae drop to the ground, where they dig in search of their hosts. It is known that second instar larvae of M. ruficauda exhibit active host searching behaviour towards its preferred host, third instar larva of Cyclocephala signaticollis. Although the means by which host location occurs has been studied and since superparasitism is a frequent scenario in the field, no information about host discrimination and host acceptance is available. We carried out studies in the field and behavioural experiments in the laboratory to determine if M. ruficauda is capable of quality host discrimination. We also studied if this parasitoid is capable of conspecific detection in order to avoid superparasitism. Finally, we analyzed the conditions under which superparasitism occurs in the field. We report here that the second instar larva of M. ruficauda is able to discriminate the parasitism status of the host by means of chemical cues, but is not capable of detecting conspecifics prior to attacking a host. We also found that the host cannot detect the presence of the parasitoid by means of chemical cues, so that no counter-defense against parasitism occurs. Furthermore, we determined that superparasitism occurs on the heavier hosts, i.e. those with more abundant resources which could harbor several parasitoid individuals. Finally, we discuss the possible implications of larval host location and host discrimination decisions on the fitness of this parasitoid.

The ontogeny of host-seeking behaviour in a parasitoid dipteran.

The robber fly Mallophora ruficauda is one of the principal apiculture pests in the Pampas region of Argentina. As adults, they prey on honeybees and other insects, while as larvae they are ectoparasitoids of third-instar scarab larvae. Females of M. ruficauda lay eggs in tall grasses. After being dispersed by the wind, larvae drop to the ground, where they dig in search of their hosts. It is known that M. ruficauda larvae exhibit active host-searching behaviour; however, it is unknown which instars are involved in this search. We carried out experiments in the laboratory to determine which larval stages are involved in host location. We report here that the second instar of M. ruficauda orientates specifically toward a source of Cyclocephala signaticollis odour, while first larval instar is indifferent to the host cues. Furthermore, we have determined that second instar larvae are more motivated to initiate exploratory movements than larva of the first stage. So far as we know, this is the first case among parasitoids, where the second instar is responsible for host location. Here we provide relevant information of this parasitoid's host-searching strategy, increasing the available knowledge of this significant apiculture pest.