Role of Cuticular Hydrocarbons in German Cockroach (Blattodea: Ectobiidae) Aggregation Behavior

Aggregation can be adaptive by providing protection from predators, facilitating thermoregulation, and expediting the location of food, shelter, and mates. German cockroaches Blattella germanica L. (Blattodea: Ectobiidae), are obligatory commensals in human-built structures, where they aggregate in crevices during the day. The source of the aggregation pheromone that drives this behavior and its chemical identity remain unclear. Cuticular hydrocarbons (CHCs) in feces have been proposed to serve as aggregation pheromone, but this function has not been investigated in relation to visual and tactile cues that mediate aggregation. Our objective was to delineate how CHCs in the feces and on the cockroach body operate in conditions that reflect the German cockroach's ecology-either applied to shelters, representing fecal deposition, or to previously extracted cockroaches, representing shelter co-habitation with other cockroaches. Cockroaches and feces-conditioned filter papers were extracted, CHCs were purified by flash chromatography, and two-choice behavior assays were performed with first instar nymphs. Our results confirmed that nymphs preferred to rest within feces-conditioned shelters. However, purified CHCs did not elicit more aggregation than solvent-treated control shelters. Nymphs significantly preferred to rest in shelters that contained a CHC-free dead female, but the addition of CHCs to the female did not enhance aggregation. Nymphs preferred to aggregate with the CHC-free female over CHC-treated shelters. Finally, a methanol extract of feces was highly effective at eliciting aggregation, contesting previous reports that fecal CHCs serve as aggregation pheromone. We assert that CHCs play a minor, if any, role in the aggregation behavior of German cockroaches.