Heat-inactivated Bifidobacterium longum ZFML006 extends lifespan in Caenorhabditis elegans via MAPK and insulin/IGF-1 signaling pathways

The global burden of aging populations and age-related functional decline presents a major public health challenge. Although live Bifidobacterium exhibits anti-aging properties, its strict anaerobic requirements limit its utility, whereas heat-inactivated preparations may offer enhanced stability and functional activity. This study characterized Bifidobacterium longum ( B. longum ) ZFML006 and systematically evaluated its anti-aging efficacy in Caenorhabditis elegans ( C. elegans ) using heat-inactivated administration. Treatment resulted in a 30.64 % increase in lifespan, a 15.20 % reduction in fecundity, a 49.92 % decrease in lipofuscin accumulation, a 20.69 % improvement in thermotolerance, a 14.8 % increase in oxidative stress survival, and enhanced motility. Additionally, it led to 10.6 % lower Reactive Oxygen Species (ROS) levels and elevated Catalase (CAT), Superoxide Dismutase (SOD), and Glutathione Peroxidase (GSH-Px) activities. Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) analysis revealed coordinated downregulation of daf-2 and akt-2 , accompanied by the upregulation of daf-16 , sod-3 , pmk-1 , nsy-1 , skn-1 , and sek-1 , confirming the modulation of both Insulin/IGF-1 signaling (IIS) and Mitogen-Activated Protein Kinase (MAPK) signaling pathways. These findings establish heat-inactivated ZFML006 as a potent anti-aging intervention by precisely regulating conserved longevity mechanisms. • Heat-inactivated form overcomes viability challenges of live Bifidobacteria , enabling shelf-stable applications in anti-aging foods and nutraceuticals. • Heat-inactivated Bifidobacterium longum ZFML006 exhibits potent antioxidant effects in Caenorhabditis elegans . • Activates longevity networks through coordinated regulation of both insulin/IGF-1 and MAPK signaling pathways.