CO₂ anesthesia on Drosophila survival in aging research

Carbon dioxide (CO₂ ) exposure is a common method of anesthesia in studies of Drosophila melanogaster. A number of negative side effects of CO₂ anesthesia have been reported. It is not clear whether the length of CO₂ anesthesia time affects Drosophila survival in aging research. Here, we examined the potential effect of the CO₂ anesthesia time length of 10-150 min. We found that long CO₂ exposure could lead to Drosophila death, more significant in males. The longer the anesthesia time is, the longer it takes for flies to wake up. Long-time CO₂ anesthesia can reduce the lifespan. Our stress tests showed that long-time CO₂ anesthesia can increase the average survival time in both males and females under starvation conditions, but can only increase female lifespan under H₂ O₂ oxidative stress. Long-time CO₂ anesthesia also significantly affects physiological traits, with spontaneous activity increased in females but decreased in males, and reduced female fecundity. Our study suggests that limiting the CO₂ anesthesia time and giving enough recovery time before performing physiological tests are important in Drosophila aging research.