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.

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.