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 Our study shows that Bd is able to infect Alerter l'administrateur Recommander à un ami Lien de l'article 

Figure 4: Intracellular colonization by Bd in zebrafish larvae.
Figure 4
(a) Zebrafish larvae bath water was inoculated with high (>200 zsp per μl) dose Bd zoospores and incubated for 72 h.p.i., then fixed and labelled with Hoechst (for DNA; blue) and mAb 5C4 (for Bd; green) for visualization by confocal microscopy. Images taken at 63 X, maximum intensity projection of Z-stack shown here. Cartoon depicts imaged region. Representative images with arrows highlight colocalization between Bd and blisters on larvae skin. Scale bar, 50 μm. (b) Zebrafish larvae bath water was inoculated with high dose Bd zoospores and incubated for 72 h.p.i., then fixed and labelled with Hoechst (for DNA; blue), mAb 5C4 (for Bd; green) and phalloidin (for F-Actin; red) for visualization by confocal microscopy. Images taken at × 63, maximum intensity projection of Z-stack shown here. Cartoon depicts imaged region. Representative images with insets highlight Bd adjacent to host cell actin rearrangements. Scale bar, 50 μm. See also Supplementary Fig. 4c for host cell actin in control treated larvae. (c) Zebrafish larvae bath water was treated as in a. Images taken at × 40 or × 63, maximum intensity projection of Z-stack shown here. Images showing different stages of Bd invasion and infection on larvae, also depicted using cartoons in the right column. 1, rhizoid-like germ tube attached to encysting zoospore, 2, chytrid thallus growth on zebrafish larvae skin, 3, encysted sporangium amongst hyperplasic epithelial cells. Scale bars, 10 μm.
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Discussion
Our study shows that Bd is able to infect and multiply on zebrafish larvae treated with antibiotics in a dose dependent manner that mimics the process of infection seen in amphibians. This demonstration of a non-amphibian vertebrate host being infected by Bd widens the host range previously known to be exploited by this hypervirulent chytrid lineage. Using a Bd monoclonal antibody (mAb 5C4), we were able to image the different stages of Bd infection with unprecedented resolution in vivo. Collectively, these results validate zebrafish larvae as a powerful aquatic model system within which these host-Bd interactions can be more fully explored. Furthermore, our observations that treating zebrafish with antibiotics results in higher cable accessories burdens of infection highlight the use of probiotic bacteria to combat Bd infection34.

Although the specific host factors necessary for Bd infection remain to be discovered, Bd is commonly found to parasitize the keratinized tissue of both amphibian and non-amphibian hosts8,9,35,36. Consistent with these observations, we found Bd parasitizing zebrafish larvae structures known to express high levels of keratin, such as the edges of the caudal fin32, where we observed fin erosion, tissue damage and apoptotic cells. These observations are in agreement with studies showing widespread apoptosis of amphibian skin cells in response to Bd infection6,31. However, in common with many macroparasite disease systems37, not all larvae became infected and mortality is heterogeneous among experiments (Supplementary Table 1), suggesting that there are unknown factors underlying the susceptibility of zebrafish larvae to Bd.

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