The robber fly Mallophora ruficauda is one of the most important apicultural pests in the Pampas region of Argentina. This species is a parasitoid of scarab beetle larvae. Females lay eggs away from the host, and the larvae perform active search behaviour toward Cyclocephala signaticollis third instar larvae, parasitoid's preferred host. This behaviour is mediated by host-related chemical cues produced in hosts’ fermentation chamber. Also, C. signaticollis larvae are attracted to fermentation chamber extracts. As scarab larvae have microbe-rich fermentation chamber, it has been suggested that microorganisms could be involved in the production of these semiochemicals. The aims of this work were first to ascertain the presence of microorganisms in the fermentation chamber of C. signaticollis larvae and second to determine the role of microorganisms in the orientation response of parasitoid and host larvae. We found that microorganisms-free C. signaticollis larvae showed deterioration in their development and did not produce the attractive semiochemicals. Therefore, we isolated fermentation chamber microorganisms of host larvae by means of different cultures media, and then, assayed different microorganisms’ stimuli by binary choice tests. We were able to isolate microorganisms and determine that M. ruficauda larvae are attracted to semiochemicals from protein degradation in the fermentation chamber. However, C. signaticollis larvae were not attracted to any semiochemicals associated with microorganisms’ activity in the fermentation chamber. Although we were unable to elucidate the exact role of gut microorganisms in host behaviour, we discuss their relevance in parasitoid host-seeking behaviour and host conspecific interaction in M. ruficauda–C. signaticollis system.

Larvae of the robber fly Mallophora ruficauda are ectoparasitoids of white grubs and adults are an important apiculture pest in Argentina. Females oviposit on tall grasses and the second instar larva actively searches and locates hosts. There are nine potential hosts in the distribution area of this parasitoid and Cyclocephala signaticollis (Coleoptera: Scarabaeidae) is the most parasitized in the field. However, M. ruficauda has a certain degree of behavioural flexibility towards different host species, and not being a strict specialist. The conditions under which the parasitoid orientates and accepts different hosts’ species are unknown. We studied the host specificity of M. ruficauda towards three species of Cyclocephala genus and we determined whether this specificity depends on larval age. We also evaluated whether larva orientation towards Cyclocephala species changes with chemical cue concentration. We assessed host specificity measuring the orientation and acceptance behaviours towards kairomones extracts and live individuals of Cyclocephala species using M. ruficauda larvae of low and high life expectancy (i.e., young and aged second instar larvae). We observed that young larvae orientated only towards C. signaticollis chemical stimulus, whereas aged larvae orientated also towards C. modesta, and the same was observed with increasing stimuli's concentration. Both young and aged M. ruficauda larvae orientate towards live C. signaticollis and C. putrida species and rejected C. modesta. Also, we found that larvae accepted all Cyclocephala hosts. In conclusion, our results indicate that specificity in the laboratory, observed through host orientation and host acceptance behaviours, depends not only on the availability of host species, but also on the nature of the host's stimuli combined with parasitoid age.

The robber fly Mallophora ruficauda is one of the principal apicultural pests in the Pampas region of Argentina. As adults, the flies prey on honey bees and other insects; while, as larvae, they parasitize 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, using host-related chemical cues. Furthermore, previous works show that these chemical cues are produced in the posterior body half of hosts. However, the precise anatomical origin of these cues and whether they mediate any behaviour of C. signaticollis larvae remains yet unknown. In order to determine the precise origin of the chemical cue, we carried out olfactometer assays with different stimuli of extracts of the posterior C. signaticollis body half. Additionally, we tested whether C. signaticollis is attracted to any of the same extracts as in the previous experiments. We found that both second instar of M. ruficauda and third instar of C. signaticollis are attracted to extracts of the fermentation chamber (proctodeum). This is the first report of attraction of conspecific larvae in scarab beetles. We discuss a possible case of system communication exploitation in an immature parasitoid-host system.