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Summary The anthropogenic spread of disease from captive to wild amphibian populations (referred to as spillover) is linked to global amphibian declines. Disinfecting procedures and protocols exist to mitigate pathogen transmission to and within natural areas, but understanding of visitor attitudes and behaviour regarding their adoption is limited. We surveyed visitors in two natural areas in a global amphibian biodiversity hotspot to assess their attitudes regarding pathogen spread in such areas and analysed the factors influencing their behavioural intentions to take specific actions to prevent pathogen spillover. Visitors’ willingness to take action was influenced by their attitudes, behavioural control and trust in wildlife/land managers, whereas socio-demographic characteristics were less influential. These findings help us to understand visitor behaviour with respect to amphibian biosecurity in natural areas and inform enhanced biosecurity measures and strategic messaging to reduce pathogen spillover. 

Abstract Batrachochytrium salamandrivorans ( Bsal ) is a fungal pathogen of amphibians that is emerging in Europe and could be introduced to North America through international trade or other pathways. To evaluate the risk of Bsal invasion to amphibian biodiversity, we performed dose-response experiments on 35 North American species from 10 families, including larvae from five species. We discovered that Bsal caused infection in 74% and mortality in 35% of species tested. Both salamanders and frogs became infected and developed Bsal chytridiomycosis. Based on our host susceptibility results, environmental suitability conditions for Bsal , and geographic ranges of salamanders in the United States, predicted biodiversity loss is expected to be greatest in the Appalachian Region and along the West Coast. Indices of infection and disease susceptibility suggest that North American amphibian species span a spectrum of vulnerability to Bsal chytridiomycosis and most amphibian communities will include an assemblage of resistant, carrier, and amplification species. Predicted salamander losses could exceed 80 species in the United States and 140 species in North America. 

Introduction One of the most important emerging infectious diseases of amphibians is caused by the fungal pathogen Batrachochytrium salamandrivorans (Bsal) . Bsal was recently discovered and is of global concern due to its potential to cause high mortality in amphibians, especially salamander species. To date, little has been reported on the pathophysiological effects of Bsal ; however, studies of a similar fungus, B. dendrobatidis (Bd) , have shown that electrolyte losses and immunosuppression likely play a key role in morbidity and mortality associated with this disease. The goal of this study was to investigate pathophysiological effects and immune responses associated with Bsal chytridiomycosis using 49 rough-skinned newts ( Taricha granulosa ) as the model species. Methods Taricha granulosa were exposed to a 1 × 10 7 per 10 mL dose of Bsal zoospores and allowed to reach various stages of disease progression before being humanely euthanized. At the time of euthanasia, blood was collected for biochemical and hematological analyses as well as protein electrophoresis. Ten standardized body sections were histologically examined, and Bsal -induced skin lesions were counted and graded on a scale of 1–5 based on severity. Results Results indicated that electrolyte imbalances and dehydration induced by damage to the epidermis likely play a major role in the pathogenesis of Bsal chytridiomycosis in this species. Additionally, Bsal -infected, clinically diseased T. granulosa exhibited a systemic inflammatory response identified through alterations in complete blood counts and protein electrophoretograms. Discussion Overall, these results provide foundational information on the pathogenesis of this disease and highlight the differences and similarities between Bsal and Bd chytridiomycosis. 

The emerging fungal amphibian pathogen, Batrachochytrium salamandrivorans (Bsal), is currently spreading across Europe and given its estimated invasion potential, has the capacity to decimate salamander populations worldwide. Fungicides are a promising in situ management strategy for Bsal due to their ability to treat the environment and infected individuals. However, antifungal drugs or pesticides could adversely affect the environment and non-target hosts, thus identifying safe, effective candidate fungicides for in situ treatment is needed. Here, we estimated the inhibitory fungicidal efficacy of five plant-derived fungicides (thymol, curcumin, allicin, 6-gingerol, and Pond Pimafix®) and one chemical fungicide (Virkon® Aquatic) against Bsal zoospores in vitro. We used a broth microdilution method in 48-well plates to test the efficacy of six concentrations per fungicide on Bsal zoospore viability. Following plate incubation, we performed cell viability assays and agar plate growth trials to estimate the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of each fungicide. All six fungicides exhibited inhibitory and fungicidal effects against Bsal growth, with estimated MIC concentrations ranging from 60 to 0.156 μg/mL for the different compounds. Allicin showed the greatest efficacy (i.e., lowest MIC and MFC) against Bsal zoospores followed by curcumin, Pond Pimafix®, thymol, 6-gingerol, and Virkon® Aquatic, respectively. Our results provide evidence that plant-derived fungicides are effective at inhibiting and killing Bsal zoospores in vitro and may be useful for in situ treatment. Additional studies are needed to estimate the efficacy of these fungicides at inactivating Bsal in the environment and treating Bsal-infected amphibians. 

Pathogen transmission through wildlife trade has become a significant One Health issue, but businesses involved in trade can take actions to minimize pathogen spread within and beyond trade networks. Such actions could include an industry‐led clean‐trade certification program whereby retailer costs for enhanced biosecurity and disease‐free product certification are offset by increased prices to consumers. However, we currently know little about the demand for, and value of such a program to consumers. With the case of pet amphibian owners in the United States, we assessed what characteristics make people more likely to demand certified wild animals and how much more in price premium they are willing to pay for such animals. Findings suggested that the demand for pathogen‐free amphibians was driven by the perceived risk of pathogen spillover to wild populations, behavioral control in preventing transmission, and other characteristics related to pet ownership and demographics. On average, respondents were willing to pay $38.65 per animal more for certified pathogen‐free amphibians than non‐certified amphibians. Findings lend support for the viability of an industry‐led clean‐trade program aimed at enhancing animal well‐being, increasing customer satisfaction, and reducing the risk of pathogen transmission within and beyond the pet trade network.

 

The recent decline in wild populations of amphibians worldwide coincides with the period of rapid growth in the global trade of wildlife. The potential for pathogen transmission within and beyond the pet amphibian trade network makes it important to explore the attitudes and behavior of businesses involved in the industry. We surveyed US businesses involved in the pet amphibian trade industry to characterize their attitudes and behaviors and identify business characteristics that could influence percieved risk of pathogen transmission in trade. We found that amphibian businesses acquire their animals from a variety of sources (e.g., importers, wholesalers, retailers, breeders, hobbyists, wild), are aware of the threat of emerging pathogens, and are concerned about the potential spillover of pathogens from captive to wild populations. Attitudes and behaviors of businesses toward pathogens varied among business types (e.g., size of business, the share of amphibian sales, mode of business operation). Moreover, businesses expressed a strong interest in acquiring amphibians that are free of pathogens and indicated a willingness to pay a price premium to acquire certified disease‐free animals. Our results indicate that the US pet amphibian trade industry is willing to participate in healthy (clean) trade practices and increasing product prices may be one option to compensate for expenses. A government program to support pathogen‐free certification would likely facilitate implementation.