## 3 Results  
### Effect on Longevity  
The parental generation was first exposed to BPA for 72 h to evaluate the effect of acute exposure to the compound, but although a slight decrease was observed at 10 mM, the results showed no significant changes in the mean survival of the subjects (Figure 5A). Humans, however, are continuously exposed to BPA, so chronic treatment throughout larval development was applied in the following two generations. The longevity of non-treated individuals, but whose parents underwent either acute (F\({}_{1}\)-N) or chronic treatment (F\({}_{2}\)-TN), was also evaluated (Figure 5). Even though there were no significant differences observed in the F\({}_{1}\)-N group when comparing the mean lifespan for each concentration and the control, [PERSON]'s exact test showed a significant change in the flies' age when their mortality reached 25% at 10 mM (Table 1). These results suggest that acute exposure of the parents may have affected the offspring's survival, even though they were not exposed to the compound. In the treated subjects of the F\({}_{1}\)-T, a significant decrease in mean survival was observed at 5 mM, under which conditions longevity did not exceed 23 days (Figure 5C). A significant decrease was also found in maximal longevity when the mortality reached 50% and 75%. At 10 mM, a significant change in survival was also observed when the mortality reached 75% (Table 1). When comparing F\({}_{1}\)-N with F\({}_{1}\)-T, we could see that, although the average survival of the F\({}_{1}\)-T generation was always lower than that of F\({}_{1}\)-N, both were affected, mainly at 2 mM and 5 mM, under which conditions the differences were significant (Figure 5D). Furthermore, at 5 mM, the differences were also significant when comparing the maximal longevity at 25%, 50% and 75% mortality. [PERSON]'s exact test even showed significant changes at 25% mortality for the 0.5, 2 and 10 mM concentrations, and at 75% for the 10 mM concentration (Table 2). We can thus conclude that chronic exposure to BPA during larval development exhibits greater toxicity than acute exposure, and that, since the most significant alterations in maximal longevity were observed in the early stages of life, this suggests that BPA may have a more relevant role in earlier deaths, and that greater emphasis should be placed on these deaths in future studies. Figure 5: The results of the longevity assay. (**A**) The mean survival obtained for the parental generation after acute exposure to BPA. (**B**) The mean survival obtained in F\({}_{1}\)-N after acute parental exposure. (**C**) The mean survival obtained in F\({}_{1}\)-T after chronic exposure throughout larval development and acute parental exposure. (**D**) Comparison between the mean survival obtained in F\({}_{1}\)-N and F\({}_{1}\)-T. (**E**) The mean survival obtained in F\({}_{2}\)-TN after chronic parental exposure. (**F**) The mean survival obtained in F\({}_{2}\)-TT after chronic exposure throughout larval development and chronic parental exposure. (**G**) Comparison between the mean survival obtained in F\({}_{2}\)-TN and F\({}_{2}\)-TT. (**H**) Comparison between the mean survival obtained in F\({}_{1}\)-T and F\({}_{2}\)-TT, to evaluate the effects of chronic exposure to BPA in successive generations. Statistical significance was obtained by the log-rank test, with the significance level set at 0.05. The results are plotted as mean + SEM when compared to the control (n = 20 flies/treatment). Legend: *—\(p<0.05\); *—\(p<0.01\); ***—\(p<0.005\).