Wild-type and mutant animals were synchronized by placing 20 gravid adult worms on NGM plates seeded with E. coli OP50 and allowing them to lay eggs for 45 min at 25°C. The gravid adult worms were then removed, and the eggs were allowed to hatch and grow at 20°C until they reached the young adult stage. The synchronized worms were transferred to NGM plates fully seeded with P. aeruginosa for 24 hours at 25°C. Worms were observed under a Leica M80 microscope with a 60× magnification at room temperature. For each worm, an average of 10 intervals between two defecation cycles were measured. The defecation cycle was identified as a peristaltic contraction beginning at the posterior body of the animal and propagating anteriorly followed by feces expulsion.

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