Antibiotic treatment and supplemental hemin availability affect the volatile organic compounds produced by P. gingivalis in vitro

Abstract

We have measured the changes in the production of volatile organic compounds (VOCs) by the oral pathogen Porphyromonasgingivalis, when treated in vitro with the antibiotic amoxicillin. We have also measured the VOC production of P.gingivalis grown in the presence and absence of supplemental hemin. Planktonic bacterial cultures were treated with different amounts of amoxicillin in the lag phase of the bacterial growth. Planktonic bacteria were also cultured with and without supplemental hemin in the culture medium. Concentrations of VOCs were measured with proton-transfer-reaction time-of-flight mass spectrometry (PTR-ToF–MS) and further molecular identification was done with gas chromatography–mass spectrometry (GC–MS) using solid phase microextraction (SPME) for sampling. The cell growth of P.gingivalis in the cultures was estimated with optical density measurements at the wavelength of 600 nm (OD600). We found that the production of methanethiol, hydrogen sulfide and several short- to medium-chain fatty acids was decreased with antibiotic treatment using amoxicillin. Compounds found to increase with the antibiotic treatment were butyric acid and indole. In cultures without supplemental hemin, indole and short- to medium-chain fatty acid production was significantly reduced. Acetic acid production was found to increase when supplemental hemin was not available. Our results suggest that the metabolic effects of both antibiotic treatment and supplemental hemin availability are reflected in the VOCs produced by P.gingivalis and could be used as markers for bacterial cell growth and response to threat. Analysis of these volatiles from human samples, such as the exhaled breath, could be used in the future to rapidly monitor response to antibacterial treatment.