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.

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.

Olfactory attraction of the larval parasitoid, Hyposoter horticola, to plants infested with eggs of the host butterfly, Melitaea cinxia.

Parasitoids locate inconspicuous hosts in a heterogeneous habitat using plant volatiles, some of which are induced by the hosts. Hyposoter horticola Gravenhost (Hymenoptera: Ichneumonidae) is a parasitoid of the Glanville fritillary butterfly Melitaea cinxia L. (Lepidoptera: Nymphalidae). Melitaea cinxia lays eggs in clusters on leaves of Plantago lanceolata L. (Lamiales: Plantaginaceae) and Veronica spicata L. (Lamiales: Plantaginaceae). The parasitoid oviposits into host larvae that have not yet hatched from the egg. Thus, though H. horticola is a parasitoid of Melitaea cinxia larvae, it must find host eggs on plants that have not been fed on by the larvae. Using a Y-tube olfactometer, the response of H. horticola to odors of Melitaea cinxia and extracts of the attacked plant species were tested. Three week-old eggs (near hatching) were attractive to young H. horticola, but one week-old eggs were attractive only to old or experienced H. horticola. Melitaea cinxia larvae were not attractive. A water extract of P. lanceolata was attractive, but ethanol or hexane extracts were not. None of the extracts of V. spicata were attractive. Leaves of V. spicata were attractive only if harboring eggs, but P. lanceolata leaves with eggs were not. Free flying H. horticola in a large outdoor enclosure were presented with host and plant cues. As in the olfactometer, V. spicata was attractive only when eggs were on it, and P. lanceolata was somewhat attractive with or without eggs. This study shows for the first time that a parasitoid of larvae uses egg volatiles or oviposition-induced plant volatiles, to find host larvae, and that Melitaea cinxia eggs or traces of oviposition induce the production of these volatiles by the plant. Based on the results, and given the natural distribution of the plants and M. cinxia eggs, parasitism of Melitaea cinxia eggs on P. lanceolata would be expected to be low. Instead, under natural conditions, a fraction of the eggs in virtually all egg clusters are parasitized on both plant species. The mismatch between the experimental results and the natural pattern of host-parasitoid interactions is discussed in terms of the expected coupling foraging cues with foraging success.

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.

Endothermy in the temperate scarab Cyclocephala signaticollis.

The increase in body temperature over that of the environment has been frequently reported in insects, in particular in relation with flight activity. Scarab beetles of the genus Cyclocephala living in tropical areas are known to exploit the heat produced by thermogenic plants, also producing heat by endothermy. Here, we report the first case of endothermy in a species of this genus living in a temperate region, Cyclocephala signaticollis. We characterised the phenomenon in this beetle using infrared thermography and exposing them to different thermal conditions. We evaluated the frequency of endothermic bouts, the nature of their periodic occurrence and their association with the activity cycles of the beetles. We found that endothermy occurs in both males and females in a cyclic fashion, at the beginning of the night, around 21:00 local time. The mean temperature increase was of 9 °C, and the mean duration of the bouts was 7 min. During endothermic bouts, the temperature of the thorax was on average 3.6 °C higher than that of the head and 4.8 °C above that of the abdomen. We found no differences between females and males in the maximum temperature attained and in the duration of the endothermy bouts. The activity period of the beetles extends throughout the whole night, with maximum activity between 22:00 and 23:00. By subjecting the beetles to different light regimes we were able to determine that the rhythm of endothermy is not controlled by the circadian system. Finally, we experimentally tested if by performing endothermy the scarabs try to reach a particular body temperature or if they invest a given amount of energy in heating up, instead. Our results indicate that at lower ambient temperature beetles show higher increase in body temperature, and that endothermy bouts last longer than at relatively higher ambient temperatures. We discuss our findings in relation to the ecology and behaviour of this beetle pest.

Host-seeking behavior in larvae of the robber fly Mallophora ruficauda (Diptera: Asilidae).

The robber fly Mallophora ruficauda is the most important pest of apiculture in the Pampas region of Argentina. Adults prey on honeybees and other insects, while larvae parasitize larvae of scarab beetles, which live underground. Females of M. ruficauda do not search for hosts but instead lay eggs in tall pastures. Once hatched, larvae drop to the ground and burrow underground to search for their hosts. We tested in the laboratory whether larvae of M. ruficauda actively search for their hosts using host and/or host-related chemical cues. We report that M. ruficauda detects its host using chemical cues that originate in the posterior half of the host's body, most likely from an abdominal exocrine structure. This particular host-searching strategy is described for the first time in Asilidae.

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.