A simplified intermittent fasting regimen robustly extends C. elegans lifespan without FUdR or antibiotic confounds

Fasting-based interventions are conserved regulators of aging that extend lifespan across species, including Caenorhabditis elegans . However, existing fasting studies are confounded by FUdR, antibiotics, germline-less mutants, and bacterial killing, which independently affect lifespan. FUdR alters lifespan, pathology, and stress responses, while antibiotics may affect worm physiology. To minimize these confounders, we developed an adult-onset intermittent fasting paradigm devoid of these factors. Wild-type worms subjected to daily fasting periods of 5 h, 6 h, or 18 h until day 10 of adulthood showed 24-57% lifespan extension. This tractable paradigm underscores the importance of limiting pharmacological and microbial conditions in dietary-intervention experiments.