R. flavipes termite colonies were collected from West Lafayette,
IN and maintained in the laboratory with 24 h of darkness on a diet
of pine wood shims and brown paper towels. Three individual
colonies were used as biological replicates in all studies. Large
termite workers (third instar or later) were used in this study;
workers lacked wing-buds and large mandibles. Sixty termites
were placed in small, Petri dishes (Nunc, 33 mm) sanded with 200
grit sandpaper. Theywere fed with a ~1 cmdisk of diet consisting of
pine wood sawdust and shredded brown paper towel (50/50 w/w).
Initially, the dietwas treated with 200 mL of one of 5 solutions based
on treatment group: water (untreated group), 5% ampicillin (w/v),
5% kanamycin (w/v), 5% metronidazole (w/v), or 2.5% tetracycline
(w/v). These concentrations were determined to be sublethal to
termites based on extensive preliminary optimization studies. Diet
disks were rewetted every other day with 100 mL of the appropriate
solution for a total holding period of seven days. After the seven day
holding period, whole guts were dissected from the termites in
sodium phosphate buffer (0.05 M, pH 7.0).