Untargeted metabolomics description of propolis's in vitro antibacterial mechanisms against Clostridium perfringens

Propolis is a natural resinous substance that is collected by honeybees (Apis mellifera) with promising antibacterial effects. Here, we examined the antibacterial activity of Chinese propolis against Clostridium perfringens, a bacterial pathogen that threatens food safety and causes intestinal erosion. The inhibitory effects of the ethanolic extract of Chinese propolis (CPE) on human-associated C. perfringens strains were determined by using the circle of inhibition, the minimum inhibitory concentrations, and bactericidal concentrations. CPE also induced morphological elongation, bacterial cell wall damage, and intracellular material leakage in C. perfringens. Untargeted HPLC-qTOF-MS-based metabolomics analysis of the bacterial metabolic compounds revealed that propolis triggered glycerophospholipid metabolism, one carbon pool by folate, and d-glutamine and d-glutamate metabolism alterations in C. perfringens. Finally, caffeic acid phenethyl ester was identified as the key active ingredient in CPE. This study suggested the usage of propolis as an alternative to antibiotics in controlling C. perfringens.