In vivo efficacy of trans-cinnamaldehyde, carvacrol, and thymol in attenuating Listeria monocytogenes infection in a Galleria mellonella model

Listeria monocytogenes is a major foodborne pathogen that causes life-threatening illnesses in humans. With emergence of antibiotic resistance in L. monocytogenes, there is considerable interest in testing the efficacy of alternative therapies for controlling listeriosis in humans. This study investigated the efficacy of three phytochemicals, namely trans-cinnamaldehyde (TC), carvacrol (CR), and thymol (TY) in reducing L. monocytogenes virulence in the recently established invertebrate model, Galleria mellonella. In addition, the effect of phytochemicals on the transcription of antimicrobial peptide genes in G. mellonella (responsible for host defense) was investigated using real-time quantitative polymerase chain reaction. G. mellonella larvae were inoculated with L. monocytogenes (10(5) CFU/larvae) either with or without the subinhibitory concentration (chemical concentration not inhibiting bacterial growth) of phytochemicals. The larvae were incubated at 37 A degrees C for 5 days, and their mortality was scored at 24-h intervals. The transcriptional response of the defense genes was studied in inoculated and uninoculated larvae at 6 h post challenge. The experiments were repeated at least six times with replicates. All phytochemicals enhanced the survival rates of G. mellonella infected with lethal doses of L. monocytogenes (P < 0.05). CR and TC at 0.01 % concentration were found to be the most effective treatments, and increased larval survival rates by 80 % and 50 %, respectively, on day 5 (P < 0.05). The phytochemicals also upregulated the expression of antimicrobial peptide genes in G. mellonella larvae challenged with L. monocytogenes (P < 0.05). Results suggest that TC, CR, and TY could potentially be used to control listeriosis. Further investigation in an appropriate mammalian model is warranted.