Figure 6 shows the M-mode contraction pattern, in which we can observe the phenomenon of prolonged relaxation time and irregular heart rhythms that occur with increasing age. In particular, flies in their ninth week of age displayed various forms of arrhythmia. Figures 7 to 10 summarize eight heartbeat parameters of male Drosophila derived and quantified by our automated algorithm, in their first, third, fifth, and ninth week, respectively. Data points represent the mean [±standard error of the mean (SEM)] for 10 files per datum point. Figure 7 shows a significant difference both in HP and HR between one- and three-week-old [analysis of variance (ANOVA), )], one- and five-week-old (ANOVA, ), one- and nine-week-old (ANOVA, ), three- and five-week-old (ANOVA, ), and three- and nine-week-old flies (ANOVA, ). It was also quite clear from our data (Fig. 8) that DI in particular increased with age, as compared to SI, with a significant difference in DI between three- and five-week-old flies (ANOVA, ). Moreover, the variation in DI was more obvious when comparing flies in their fifth and ninth weeks; this was consistent with Fig. 9, which shows the average AI, reflecting the age-related increase in arrhythmicity in flies (ANOVA, ). However, the average size for ESA remains similar across age, while a small but statistically insignificant increase in EDA is shown in Fig. 10(a). For this reason, no significant decline in FS was observed, as shown in Fig. 10(b).