Potential Protective Role of Amphibian Skin Bacteria Against Water Mold Saprolegnia spp.

Amphibian populations have experienced a severe decline over the past 40 years, driven primarily by environmental pollution, habitat destruction, climate change, and disease. This work reports, for the first time, saprolegniosis in <i>Pelophylax perezi</i> egg masses and saprolegniosis in amphibians in Portugal. After isolation and phylogenetic analysis, the pathogen was identified as <i>Saprolegnia australis</i>. Following this, the present work intended to screen a collection of <i>P. perezi</i> skin bacteria for the existence of bacterial strains with inhibitory action against the newly identified <i>S. australis</i> SC1 and two other species, <i>Saprolegnia diclina</i> SAP 1010 UE and <i>Saprolegnia australis</i> SAP 1581 UE. The results showed that various bacterial species could inhibit the growth of these three species of oomycetes. Bacteria with the most significant antagonistic action against <i>Saprolegnia</i> spp. predominantly belonged to the genus <i>Bacillus</i>, followed by <i>Serratia</i>, <i>Pseudomonas</i>, and <i>Aeromonas</i>. Despite variations in bacterial diversity among frog populations, the present study also demonstrated the presence of bacteria on frogs' skin that were capable of inhibiting <i>Saprolegnia</i> spp., as evidenced by <i>in vitro</i> challenge assays. These findings highlight the protective function of bacteria present in amphibian skin. The observed bacterial diversity may contribute to the metabolic redundancy of the frog skin microbiome, helping to maintain its functional capacity despite shifts in the community composition. Additionally, the study found that, when providing a more advantageous environment for pathogen growth-in this case a peptone-glucose (PG) medium instead of R2A-the percentage of bacteria with moderate-to-strong antagonistic activity dropped by 13% to 4%. In conclusion, the presence of bacteria capable of inhibiting <i>Saprolegnia</i> spp. in adult individuals and across different environmental conditions may contribute to lowering the susceptibility of frog adults towards <i>Saprolegnia</i> spp., compared with that in the early stages of development, like the tadpole or egg stages.