Comparing electroantennogram and behavioral responses of two Pseudacteon phorid fly species to body extracts of Black, Red and Hybrid imported fire ants, Solenopsis spp.

Several phorid fly species were introduced to the southern United States for biological control of the invasive imported fire ants, Solenopsis richteri (Black), Solenopsis invicta (Red), and their Hybrid S. richteri×S. invicta (Hybrid). It has been previously reported that the Jaguariuna biotype of Pseudacteon tricuspis and the Formosan biotype of Pseudacteon curvatus could distinguish among the three fire ant species with greater preference for Hybrid and Red fire ants. We hypothesized that phorid flies might use host derived chemical cues to differentiate ant species. To determine possible differential olfactory sensitivity of phorid fly species to different fire ant species, we compared electroantennogram (EAG) and behavioral responses of both sexes of P. tricuspis and P. curvatus to body extracts of Black, Red and Hybrid fire ants. As worker sizes of Black and Hybrid fire ants used in this study were much larger than that of Red fire ant (the average weight for Black, Red and Hybrid workers was 1.707, 0.863, 1.223mg per ants, respectively), at doses of 0.01, 0.1, 1 worker equivalent, body extracts of Black and Hybrid fire ant elicited significantly greater EAG response in both sexes of P. tricuspis than that of Red fire ant. Similarly, the EAG response in female P. curvatus to body extract of Black fire ant was significantly greater than to body extract of Red fire ant. To eliminate worker size influence on EAG response in phorid flies, we conducted a second EAG study using a dose of 1mg ant equivalent (body extract from 1mg of worker). No difference in EAG responses was recorded to body extract obtained from the same amount of workers among the three fire ant species (we consider viable Hybrid fire ant as a species in this paper), suggesting that worker size differences contributed to difference in EAG response in the first EAG study. In both EAG studies, male P. tricuspis showed significantly greater EAG responses than male P. curvatus to all three fire ant species. In four-way olfactometer bioassay, worker body extracts of all three fire ant species were equally attractive to P. tricuspis and P. curvatus (i.e. both phorid fly species did not show any preferences among the three fire ant species). Together, the results of the EAG and behavior studies suggest that parasitic phorid flies utilize host derived non-polar compounds from worker ants extracted out by hexane for host location but not for host preference, since both fly species are not able to distinguish among the body extracts of the three fire ant species. Future study will investigate possible involvement of polar compounds and/or non-chemical cues in mediating host preference by phorid flies.