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  1. Reguera, Gemma (Ed.)
    ABSTRACT Mucosal defenses are crucial in animals for protection against pathogens and predators. Host defense peptides (antimicrobial peptides, AMPs) as well as skin-associated microbes are key components of mucosal immunity, particularly in amphibians. We integrate microbiology, molecular biology, network-thinking, and proteomics to understand how host and microbially derived products on amphibian skin (referred to as the mucosome) serve as pathogen defenses. We studied defense mechanisms against chytrid pathogens, Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal), in four salamander species with different Batrachochytrium susceptibilities. Bd infection was quantified using qPCR, mucosome function (i.e., ability to kill Bd or Bsal zoospores in vitro ), skin bacterial communities using 16S rRNA gene amplicon sequencing, and the role of Bd-inhibitory bacteria in microbial networks across all species. We explored the presence of candidate-AMPs in eastern newts and red-backed salamanders. Eastern newts had the highest Bd prevalence and mucosome function, while red-back salamanders had the lowest Bd prevalence and mucosome function, and two-lined salamanders and seal salamanders were intermediates. Salamanders with highest Bd infection intensity showed greater mucosome function. Bd infection prevalence significantly decreased as putative Bd-inhibitory bacterial richness and relative abundance increased on hosts. In co-occurrence networks, some putative Bd-inhibitory bacteria were found as hub-taxa, with red-backs having the highest proportion of protective hubs and positive associations related to putative Bd-inhibitory hub bacteria. We found more AMP candidates on salamanders with lower Bd susceptibility. These findings suggest that salamanders possess distinct innate mechanisms that affect chytrid fungi. IMPORTANCE How host mucosal defenses interact, and influence disease outcome is critical in understanding host defenses against pathogens. A more detailed understanding is needed of the interactions between the host and the functioning of its mucosal defenses in pathogen defense. This study investigates the variability of chytrid susceptibility in salamanders and the innate defenses each species possesses to mediate pathogens, thus advancing the knowledge toward a deeper understanding of the microbial ecology of skin-associated bacteria and contributing to the development of bioaugmentation strategies to mediate pathogen infection and disease. This study improves the understanding of complex immune defense mechanisms in salamanders and highlights the potential role of the mucosome to reduce the probability of Bd disease development and that putative protective bacteria may reduce likelihood of Bd infecting skin. 
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  2. Abstract

    We performed 3 experimental reintroductions of the critically endangered Wyoming toad (Anaxyrus baxteri) to study the toad's spatial ecology, behavior, and test the effectiveness of a soft‐release strategy at Mortenson Lake National Wildlife Refuge, Wyoming, USA. We tracked reintroduced, captive‐bred toads (n = 46) and field collected toads that had been in the wild for at least one year (n = 12) using a harmonic tracking system totaling 1,438 toad relocations in 2014 and 2015. We compared the spatial ecology and behavior of soft‐released toads, hard‐released toads, and overwintered toads. Hard‐released toads were transported to the reintroduction site and released. Soft‐released toads were kept in an outdoor enclosure at the release site for 14 days before release. Compared to hard‐released toads, the soft‐release group's average total path distance moved was lower (mean difference = 263.23, CI = 115.37 – 411.05,P= 0.007). Additionally, hard‐released toads in the first 2 days post release moved 380% farther than the soft‐released toads in the same 2‐day period. Our results indicated that soft‐releases are an effective way to improve site fidelity and retain wild‐type spatial ecology and behavior of reintroduced captive‐bred amphibians.

     
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