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Fig. 2
Symptoms of Bd infection in zebrafish larvae.
(a) Zebrafish larvae bath water was inoculated with mTGhL plate washings (control, open circles), low (<200 zsp per μl, filled circles) or high (>200 zsp per μl, filled squares) dose Bd zoospores. Larvae were incubated for 72 h.p.i., where dorsal and ventral fin length was measured (as depicted by red line in cartoon above each graph). Each point represents an individual larva. Data pooled from three experiments per dose, using n=4–12 per treatment. Mean±s.e.m. are shown. Significance testing performed using unpaired Student’s t-test (two-tailed), ns P>0.05, ***P<0.001. (b) Zebrafish larvae bath water was inoculated with control, low or high dose Bd zoospores, incubated for 72 h.p.i. and imaged by stereomicroscopy. Representative images with cyan outline showing presence or erosion of fin over time. (c) Zebrafish larvae bath water was inoculated with control, low or high dose Bd zoospores and incubated for 24, 48 or 72 h.p.i., then labelled with calcofluor white (CFW; for chitin, green) and imaged by fluorescent stereomicroscopy. Representative images with dotted outline of larvae showing (CFW)-labelled punctae on infected larvae. (d) Enumeration of CFW-labelled punctae from zebrafish larvae whose bath water was inoculated with control (open circles), low (filled circles) or high (filled squares) dose Bd zoospores and incubated for 72 h.p.i. Each point represents an individual larva. Data pooled from three experiments per dose, using n=12 per treatment. Mean±s.e.m. are shown. Significance testing performed using Mann–Whitney test (two tailed), ns P>0.05, ***P<0.001. See also Supplementary Fig. 2e for replicate experiments. (e) Zebrafish larvae bath water was inoculated with high dose Bd zoospores and incubated for 72 h.p.i., then labelled with CFW (green) and imaged by fluorescent stereomicroscopy. Representative images with insets showing colocalization of CFW-labelled punctae with blisters.