1A). If they approached the access point but did not attempt or complete the worm pulling, it was still considered orientation time. Manipulation index is defined as the duration after the animal starts to reach in through the access point until the time the food reward is pulled. Orientation time for sighted crayfish in both white and red light and blind crayfish in red light indicated a significant effect from the first day and last day

of the experiment using the repeated measures ANOVA (F4,79 = 12.288, P < 0.001; using a Bonferroni cutoff of P < 0.0035 to account for a family-wise error rate of P < 0.05) with sizeable variation among the crayfish (residual standard Inhibitors,research,lifescience,medical deviation of 1.12 for log(Time) with the standard deviation across

crayfish of 0.84; Fig. 4I). Figure 4 Graphical representation of orientation and manipulation times for both species in white and red light. (A) Sighted crayfish in white light showing individual minutes to orient and locate access point. (B) Sighted crayfish in white light showing individual … In contrast, blind Inhibitors,research,lifescience,medical crayfish in white light did not show a significant difference from the first day to the last day of the experiment (F4,79 = 12.288, Inhibitors,research,lifescience,medical P < 0.028; using a Bonferroni cutoff of P < 0.0035 to account for a family-wise error rate of P < 0.05). Thus, there was a significant difference between the other experimental groups (sighted white/red light, blind red light) from Day 1 to Day 38, but not for blind crayfish in white light (Fig. 4I). Significance is shown on the graph with boxes representing the points of statistical comparison. Furthermore, group values on Day 38 were statistically tested across

groups and check details showed no significance between sighted red/white light Inhibitors,research,lifescience,medical and blind red light, but there is a significant difference to blind white light (shown on graph). To understand the actual time spent completing the motor task, Inhibitors,research,lifescience,medical analysis of manipulation index separated out the actual time between access point location and when the first worm was pulled. The manipulation index analysis indicated a significant effect for all experimental groups when comparing Day 1 and Day 38 (Fig. 4J). Specifically, the repeated measures ANOVA comparison for sighted crayfish in white and red light, and blind crayfish in white and red light, showed a significant difference from the first day of learning (F4,79 = 5.78, P < 0.001) and no significant difference from each other with sizeable variation among the crayfish (residual standard deviation of 1.42 for log(Time) with the standard deviation MycoClean Mycoplasma Removal Kit across crayfish of 0.93). Significance is shown on the graph with boxes representing the points of statistical comparison (Day 1, Day 38) as well as significance between groups on Day 38. Sighted crayfish showed more individual variability during manipulation time with white light, so the ability to see the food reward may have interfered with the ability to manipulate the cheliped in the same time frame that occurred in red light.