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Fig. 6 | BMC Biology

Fig. 6

From: Skin-penetrating nematodes exhibit life-stage-specific interactions with host-associated and environmental bacteria

Fig. 6

P. mirabilis impairs both egg and larval development in S. stercoralis. a Representative images of S. stercoralis embryonic development, a healthy pre-infective larva, and an unhealthy pre-infective larva. Egg development was classified into the categories indicated based on the classifications used for C. elegans [29]. Scale bars = 20 μm. b The percentage of unhatched eggs that fell into each category of embryonic development for worms cultured on either P. mirabilis or E. coli. For each experiment, 1 female and 3 males were cultured on the bacterial lawn for 24 h, after which the worms were removed from the plate and the percentage of eggs in each category was scored. The percentage of eggs in each category was then scored again after 48 h. After 24 h, more undeveloped eggs were observed on P. mirabilis relative to E. coli, suggesting that P. mirabilis interferes with the earliest stages of egg development (left). **p< 0.01, ns = not significant, two-way ANOVA with Sidak’s multiple comparisons post-test. n = 9–10 trials for each condition. After 48 h, nearly all of the eggs on the E. coli plates had hatched; the remaining eggs were almost all undeveloped. In contrast, approximately half of the eggs on P. mirabilis were at the pretzel stage of development while the other half were undeveloped, indicating that culturing on P. mirabilis delays or prevents egg hatching in some eggs that have initiated embryonic development (right). ***p< 0.001, ns = not significant, two-way ANOVA with Sidak’s multiple comparisons post-test. n = 6-10 trials for each condition. c. The percentage of the total progeny consisting of undeveloped eggs, developing eggs, healthy larvae, or unhealthy larvae from the experiment described in (b) after 24 h (left) or 48 h (right). Culturing on P. mirabilis resulted in an increased proportion of eggs that failed to develop and of the worms that hatched, an increased number of unhealthy larvae. Larvae that were classified as unhealthy appeared non-motile and generally remained in a curled position. *p< 0.05, ****p< 0.0001, two-way ANOVA with Tukey’s multiple comparisons test. Comparisons are between the same category and time point; only significant differences are shown. n = 7–10 trials for each condition. Median percentages are represented, and bars represent interquartile ranges. Data in (b) and (c) are from the same experiments

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