IDENTIFYING ANTIMICROBIAL MOLECULES AGAINST LEGIONELLA PNEUMOPHILA

AND OTHER BACTERIAL PATHOGENS FROM HONEY BACTERIA.

Jeffrey Jian, Carson Shin, Ahmed Warshanna, Joshua Khorsandi, Laura Sanchez-
Buitrago, Benjamin Orsburn, and Tamara J. O’Connor

Introduction: Antimicrobial resistance (AMR) is an immediate threat to human health. Over

the last several decades, many bacterial infections have become increasingly difficult, if not
impossible, to treat due to reduced antibiotic efficacy as a result of the rising incidence of
drug-resistant bacteria. This indicates an urgent need for the identification and development
of novel antibiotics. Given that many clinically-relevant antibiotics are of natural product
origin, investigating natural product sources remains critical for identifying and developing
novel therapeutics.

Methods: Honey was plated to observe inhibition of Legionella pneumophila growth.

Individual clonal isolates were cultured and sequenced using 16S RNA. L. pneumophila
was grown in either AYE culture medium or minimally-defined Ristroph medium (MRM). L.
pneumophila supernatant was harvested, filter-sterilized, and used to induce the isolated
honey bacterial species. Supernatant from the induced honey bacteria was filter-sterilized
and concentrated 10-fold. Antibacterial activity of the concentrated supernatant was used to
inhibit L. pneumophila growth via disk-diffusion assays, and zone diameters of inhibition
were measured as a readout of anti-Legionella growth.

Results: We have identified a number of previously-uncharacterized bacterial species from

raw honey that can produce and secrete molecules with antibacterial activity. The
production of these antimicrobial molecules occurs when honey bacteria are culture in the
presence of L. pneumophila, a Gram-negative pathogen that causes Legionnaires’ disease,
a life-threatening pneumonia. We have determined that the production of these molecules
occurs in response to diffusible molecules secreted by L. pneumophila. The secreted
molecules appear to be active in inhibiting both L. pneumophila and other known bacterial
species with antibacterial resistance.

Discussion: We are currently determining the identities of these honey bacteria-generated

antimicrobials molecules, their modes of action, and the sensory and biosynthetic pathways
responsible for their production upon exposure to L. pneumophila. Defining these active
molecules provides a unique opportunity for the discovery of potentially novel antibiotics to
active molecule to treat bacterial infections.