Chytrid fungal (Bd)/Ranavirus Co-infection Model using Alytes muletensis frogs: Evaluation of Host/Pathogen Interactions using the C-bellyskin Organ Culture Method

Chytrid fungal (Bd) and Ranavirus (Rv) infections can lead to catastrophic amphibian population declines. These pathogens often co-occur in amphibian populations and preliminary evidence suggests that survival is diminished if frogs are already infected with Bd prior to Rv infection. The present study aimed to uncover the host/pathogen response in the context of a co-infection model. Young metamorphs were exposed to high concentration Bd. Whole animal and C-bellyskin organ culture assays were compared as a potential method to reduce animal use. After Bd exposure, the whole animal or the organ cultured bellyskin was exposed to either heat-killed or live Rv. Using the NovaSeq6000 platform, polyA mRNA strand-specific library construction and sequencing was performed to generate 2 x 150bp paired-end RNA-seq reads (50 million reads per sample).