Synchronized fourth-stage (L4) animals (at least eight animals per strain per experiment) were singly transferred onto 60-mm NGM agar plates seeded with half the normal volume of E. coli OP50 (50 μl) to improve visibility of eggs. Animals were incubated at 20°C, and eggs laid were counted manually every day using a dissecting microscope. Animals were transferred to new plates every 24 hours until the end of their reproductive life spans. Counts for each animal were summed to determine brood size. Serial, egg-laid plates were maintained at 20°C and evaluated for the number of viable larvae compared to the number of eggs identified within each plate, as an index of viability. Experiments were replicated three times by two independent evaluators.

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