Hatching of whipworm eggs induced by bacterial contact is serine-protease dependent
Fig 5
Bacteria-mediated hatching of Trichuris muris eggs is protease-dependent.
Trichuris muris eggs were co-cultured with Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa and Staphylococcus aureus in the absence (-) or presence (+) of a protease inhibitor cocktail (1x) for A. 2 h and B. 4 h at 37°C. The number of total embryonated eggs and hatched larvae were counted, from which the hatching percentage was calculated. Hatching was completed in triplicate across three independent experiments (n = 9), except for S. typhimurium in A, where data comes from two independent experiments (n = 6). Median and interquartile range are shown, and statistical differences between the absence (-) or presence (+) of a protease inhibitor cocktail for each bacterial species were evaluated using the Mann-Whitney test (**p<0.005, ****p<0.0001). C. T. muris eggs were co-cultured with E. coli, S. typhimurium, P. aeruginosa and S. aureus in the absence (-) or presence (1x or 2x) of a protease inhibitor cocktail for 24 h at 37°C. The number of total embryonated eggs and hatched larvae were counted, from which the hatching percentage was calculated. Hatching was completed in triplicate across three independent experiments (n = 9). Median and interquartile range are shown, and statistical differences between the absence (-) or presence (1x or 2x) of a protease inhibitor cocktail for each bacterial species were evaluated using Kruskal Wallis with Dunn’s multiple comparison tests performed (*p<0.05, **p<0.005, ***p<0.0005, ****p<0.0001). D. T. muris eggs were co-cultured with E. coli in the absence (-) or presence (+) of a protease inhibitor cocktail (1x) for 24 h at 37°C. The number of total embryonated eggs and hatched larvae were counted, from which the hatching percentage was calculated. Next, T. muris eggs co-cultured in the presence of the inhibitor were washed to remove it and re-exposed to fresh E. coli cultures for 24 h at 37°C; the percentage hatching was recorded 24 h later (Washed). Hatching was completed in triplicate in the absence and presence of the protease inhibitor cocktail and sextuplicate for the washed condition across three independent experiments (n = 9 and n = 18, respectively). Median and interquartile range are shown, and statistical differences between groups were evaluated using Kruskal Wallis with Dunn’s multiple comparison tests performed (*p<0.05, **p<0.005, ****p<0.0001).