Antibacterial activity and metabolomic analysis of linalool against bovine mastitis pathogen Streptococcus agalactiae

Streptococcus agalactiae is among the major causative pathogens of bovine mastitis, as well as crucial pathogen leading to human morbidity and mortality. Being a promising natural antibacterial agent, linalool has been broadly applied in medicine and food processing. However, its antibacterial effect against S. agalactiae has barely been elucidated. This study is the first to investigate the antibacterial activity and action mechanism of linalool against S. agalactiae causing bovine mastitis. Linalool exhibited significant antibacterial activity against S. agalactiae, with an inhibition zone diameter of 23 mm and a minimum inhibitory concentration of 1.875 μL/mL. In addition, linalool damaged cell structural integrity of S. agalactiae, leading to the leakage of intracellular components (alkaline phosphatase, nucleic acids and protein). Linalool also exhibited a scavenging effect on biofilm. Moreover, untargeted metabolomics analysis revealed that linalool stress substantially disrupted intracellular metabolism of S. agalactiae. Linalool caused energy metabolism disorder, and obstructed nucleic acid synthesis in S. agalactiae. Furthermore, downregulation of amino acids (e.g., proline, alanine) and upregulation of saturated fatty acids provide strong evidence for linalool induced cell wall and membrane damage. Overall, linalool exhibited strong antibacterial activity against S. agalactiae by destroying the cell structure and disrupting intracellular metabolism. This study provides a new insight and theoretical foundation for linalool application in preventing S. agalactiae infection.