Use of genetically engineered Escherichia coli to monitor ingestion, loss, and transfer of bacteria in termites.

Escherichia coli was transformed with a recombinant plasmid (pEGFP) containing the genes for ampicillin resistance and Green Fluorescent Protein (GFP). Escherichia coli expressing GFP (E. coli/GFP+) was then fed to workers of the termite Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae). The transformed bacteria in the termite guts were detected by growing the gut flora under selective conditions and then checking the cultures for fluorescence. Recombinant plasmids in the termite gut were detected by plasmid extraction with subsequent restriction enzyme digest. The presence of the GFP gene in the gut of termites fed with E. coli/GFP+ was verified by PCR amplification. Transformed E. coli were ingested rapidly when workers fed on filter paper inoculated with E. coli/GFP+. After 1 day, 42% of termite guts harbored E. coli/GFP+. Transfer of E. coli/GFP+ from donor termites (fed with E. coli/GFP+) to recipients (fed with moist filter paper) occurred within 1 day. However, without continuous inoculation, termites lost the transformed bacteria within 1 week.