Characterization of Antibacterial Activities of Eastern Subterranean Termite, Reticulitermes flavipes, against Human Pathogens

The emergence and dissemination of multidrug resistant bacterial pathogens necessitate research to find new antimicrobials against these organisms. We investigated antimicrobial production by eastern subterranean termites, Reticulitermes flavipes , against a panel of bacteria including three multidrug resistant (MDR) and four non-MDR human pathogens. We determined that the crude extract of naïve termites had a broad-spectrum activity against the non-MDR bacteria but it was ineffective against the three MDR pathogens Pseudomonas aeruginosa , methicillin-resistant Staphylococcus aureus (MRSA), and Acinetobacter baumannii . Heat or trypsin treatment resulted in a complete loss of activity suggesting that antibacterial activity was proteinaceous in nature. The antimicrobial activity changed dramatically when the termites were fed with either heat-killed P . aeruginosa or MRSA. Heat-killed P . aeruginosa induced activity against P . aeruginosa and MRSA while maintaining or slightly increasing activity against non-MDR bacteria. Heat-killed MRSA induced activity specifically against MRSA, altered the activity against two other Gram-positive bacteria, and inhibited activity against three Gram-negative bacteria. Neither the naïve termites nor the termites challenged with heat-killed pathogens produced antibacterial activity against A . baumannii . Further investigation demonstrated that hemolymph, not the hindgut, was the primary source of antibiotic activity. This suggests that the termite produces these antibacterial activities and not the hindgut microbiota. Two-dimensional gel electrophoretic analyses of 493 hemolymph protein spots indicated that a total of 38 and 65 proteins were differentially expressed at least 2.5-fold upon being fed with P . aeruginosa and MRSA, respectively. Our results provide the first evidence of constitutive and inducible activities produced by R . flavipes against human bacterial pathogens.