Body weight
The water treatment experienced during development did not affect male body weight at emergence (mean ± SE = 3.863 ± 0.030 mg, χ2 = 0.003, Df = 1, p = 0.959, Table A1). Males’ change in body weight prior to mating was not affected by the interaction between juvenile and adult water treatment (χ2 = 1.285, Df = 1, p = 0.257, Table A2). However, both wet juvenile environments and wet adult environments caused greater weight loss before mating (juvenile: χ2 = 4.405, Df = 1, p = 0.036; adult: χ2 = 25.842, Df = 1, p < 0.001, Fig. 1a, Table A2), and males that were larger at emergence lost more weight (χ2 = 138.428, Df = 1, p < 0.001, Table A2).
The change in male weight between mating days was affected by an interaction between juvenile and adult water treatments (χ2 = 4.933, Df = 1, p = 0.026, Table A3). However, post hoc tests revealed that the dominant pattern in the data is driven by male access to water during adulthood (Fig. 1b, Table A4). Furthermore, larger males lost more weight between matings (χ2 = 5.705, Df = 1, p = 0.017, Table A3) and males lost less weight between the second and third mating day than they did between the first and second mating day (χ2 = 29.484, Df = 1, p < 0.001, Table A3).
Ejaculate traits
Neither juvenile water treatment, nor the interaction between juvenile and adult water treatment affected male ejaculate weight in their first mating (juvenile: χ2 = 0.635, Df = 1, p = 0.426, juvenile*adult interaction: χ2 = 0.009, Df = 1, p = 0.926, Table A5). However, males with access to water during adulthood had a larger ejaculate in their first mating than those without (χ2 = 4.344, Df = 1, p = 0.037, Fig. 2, Table A5), and males who were larger at emergence had larger ejaculates in their first mating (χ2 = 9.613, Df = 1, p = 0.002, Table A5).
Ejaculate replenishment across days was not affected by the three-way nor two-way interactions involving juvenile water treatment, adult water treatment or mating day (all p > 0.089, Table A6), nor was it affected by juvenile or adult water treatments (juvenile: χ2 = 0.084, Df = 1, p = 0.771; adult: χ2 = 0.068, Df = 1, p = 0.794, Table A6). Ejaculate replenishment was also not affected by male body weight at emergence (χ2 = 0.701, Df = 1, p = 0.402, Table A6). However, the difference in ejaculate weight between the first and second day of mating was greater than between the second and third day (χ2 = 267.236, Df = 1, p < 0.001, Table A6).
The change in ejaculate weight between matings on the same day was not affected by the three-way or the two-way interactions involving juvenile water treatment, adult water treatment and mating day (all p > 0.095, Table A7). There was also no effect of the juvenile water treatment (χ2 = 1.611, Df = 1, p = 0.204, Table A7), adult water treatment (χ2 = 3.244, Df = 1, p = 0.072, Table A7), or male body weight at emergence (χ2 = 0.368, Df = 1, p = 0.544, Table A7). However, there was a greater decrease in ejaculate weight on the first mating day (Table A9).
Mating behavior
When looking at mating behavior during the first mating, mating duration was not affected by the interaction between juvenile and adult water treatment (χ2 = 2.689, Df = 1, p = 0.101, Table A10) or the adult water treatment (χ2 = 0.018, Df = 1, p = 0.893, Table A10). However, males reared in wet beans had shorter matings than those reared in dry beans (χ2 = 7.158, Df = 1, p = 0.007, Fig. 3, Table A10). Also, larger males had shorter mating durations than smaller males (χ2 = 3.995, Df = 1, p = 0.046, Table A10), but female body weight had no effect (χ2 = 1.493, Df = 1, p = 0.222, Table A10).
Kicking latency, on the other hand, was affected by the interaction between juvenile and adult water treatment (χ2 = 8.044, Df = 1, p = 0.005, Table A11). Males reared in dry beans, without access to water during adulthood were kicked later than males reared in dry beans, with access to water during adulthood (t ratio = 3.115, p = 0.011, Fig. 3b, Table A12) as well as those reared in wet beans, without access to water (t ratio = 3.395, p = 0.005, Fig. 3b, Table A12). Finally, matings involving males with access to water as adults had longer kicking duration (χ2 = 6.384, Df = 1, p = 0.012, others: p > 0.133, Fig. 3c, Table A13).
When analyzing mating duration across the entire experiment, there were no two or three-way interactions (all p > 0.065, Table A14). Indicating that neither juvenile or adult water treatment mediated changes in mating duration that occurred over subsequent matings. Further, neither juvenile or adult water treatment had any overall effects on mating duration (juvenile: χ2 = 0.284, Df = 1, p = 0.593; adult: χ2 = 0.098, Df = 1, p = 0.754, Table A14). However, mating duration tended to increase with subsequent matings both within and across days. Specifically, mating duration was longer on mating days two and three than it was on mating day one (χ2 = 119.591, Df = 2, p < 0.001, Table A15), and mating duration was longer for the second mating than the first mating of each mating day (χ2 = 81.546, Df = 1, p < 0.001, Table A14). Finally, although larger males had longer matings (χ2 = 18.755, Df = 1, p < 0.001, Table A14), larger females had shorter matings (χ2 = 4.671, Df = 1, p = 0.031, Table A14).
Our analysis of kicking latency also showed no significant two or three-way interactions (all p > 0.078, Table A16), apart from a significant interaction between juvenile water treatment and mating day (χ2 = 7.694, Df = 2, p = 0.021, Table A16). This interaction reflects that for males reared in dry beans kicking latency initially increases from day one to day two and then plateaus (day 1 vs. day 2: t ratio = -7.141, p < 0.001; day 2 vs. day 3: t ratio = 0.170, p = 1.000, Fig. 4, Table A17), whereas for males reared in wet beans kicking latency increase across all three days (day 1 vs. day 2: t ratio = -7.405, p < 0.001; day 2 vs. day 3: t ratio = -3.153, p = 0.021, Fig. 4, Table A17). Further, as for mating duration, adult water treatment had no overall effect on kicking latency (χ2 = 0.621, Df = 1, p = 0.431, Table A16), but there were significant effects of mating order within each day as well as male and female size. Males were kicked later during the second mating of each day (χ2 = 120.766, Df = 1, p < 0.001, Table A16), larger males were kicked sooner than smaller males (χ2 = 11.081, Df = 1, p < 0.001, Table A16) and larger females started kicking later than smaller females (χ2 = 10.581, Df = 1, p = 0.001, Table A16).
For kicking duration, none of two or three-way interactions had a significant effect (all p > 0.068, Table A19), and there were no significant main effects (all p> 0.050, Table A19) apart from effects of mating order and male size. These showed that, males were kicked for a shorter duration in the second mating of each mating day (χ2 = 7.129, Df = 1, p = 0.008, Table A19) and that larger males generally experienced longer kicking durations than smaller males kicked longer (χ2 = 5.056, Df = 1, p = 0.025, Table A19).
Lifespan
Male lifespan was not affected by an interaction between juvenile and adult water treatment, nor the juvenile water environment on its own (mean ± SE = 13.987 ± 0.320 day, Table A20). However, males with access to water during adulthood lived significantly longer than those without (z = -3.118, p = 0.002, Fig. 5, Table A20), as did males that were larger at emergence (z = -4.214, p < 0.001, Table A20).