Taken together, our results demonstrated that thymine potentiates bactericidal antibiotics activity against Gram-negative pathogens through activating bacterial metabolism, providing a universal strategy to overcome Gram-negative pathogens. In Galleria mellonella infection model, thymine effectively improved ciprofloxacin activity against E. Subsequently, metabolically inactive bacteria were converted to active bacteria and restored its susceptibility to antibiotic killing. The mechanisms underlying this finding were that exogenous thymine could upregulate bacterial metabolism including increased TCA cycle and respiration, which thereby promote the production of ATP and ROS. Similar promotion effects were also observed for other bactericidal antibiotics, including ampicillin and kanamycin, in the fight against four kinds of Gram-negative bacteria. coli in a growth phase-independent manner. Herein, we found that thymine potentiated ciprofloxacin killing against both sensitive and resistant- E. However, the adjuvant potential of nucleotides in combination with antibiotics to kill Gram-negative pathogens remains unknown. Bacterial metabolic state has been shown to play a critical role in its susceptibility to antibiotic killing. Alternative strategies are required to combat antibiotic refractory bacteria. 4Joint International Research Laboratory of Agriculture and Agri-Product Safety, The Ministry of Education of China, Yangzhou University, Yangzhou, Chinaĭiminished antibiotic susceptibility of bacterial pathogens is an increasingly serious threat to human and animal health.3Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, China.2Institute of Comparative Medicine, Yangzhou University, Yangzhou, China.
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