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We designed two probiotic treatments to control chytridiomycosis caused by Batrachochytrium dendrobatidis (Bd) on infected Panamanian golden frogs (Atelopus zeteki), a species that is thought to be extinct in the wild due to Bd. The first approach disrupted the existing skin microbe community with antibiotics then exposed the frogs to a core golden frog skin microbe (Diaphorobacter sp.) that we genetically modified to produce high titers of violacein, a known antifungal compound. One day following probiotic treatment, the engineered Diaphorobacter and the violacein-producing pathway could be detected on the frogs but the treatment failed to improve frog survival when exposed to Bd. The second approach exposed frogs to the genetically modified bacterium mixed into a consortium with six other known anti-Bd bacteria isolated from captive A. zeteki, with no preliminary antibiotic treatment. The consortium treatment increased the frequency and abundance of three probiotic isolates (Janthinobacterium, Chryseobacterium, and Stenotrophomonas) and these persisted on the skin 4 weeks after probiotic treatment. There was a temporary increase in the frequency and abundance of three other probiotics isolates (Masillia, Serratia, and Pseudomonas) and the engineered Diaphorobacter isolate, but they subsequently disappeared from the skin. This treatment also failed to reduce frog mortality upon exposure.

A chytridiomycosis outbreak from Batrachochytrium dendrobatidis ( Bd) in a mixed-species plethodontid salamander exhibit resulted in four green salamander ( Aneides aeneus) deaths. One green salamander died before treatment, and three died during treatment with daily 0.005% itraconazole baths. All salamanders had evidence of severe Bd infections via cytology, histopathology, and/or polymerase chain reaction (PCR) at the time of death. Ten long-tailed salamanders ( Eurycea longicauda) and one two-lined salamander ( Eurycea bislineata) that shared the enclosure were initially negative for Bd on quantitative PCR but were prophylactically treated with daily 0.01% itraconazole baths for 11 days. Posttreatment testing yielded eight long-tailed salamanders and one two-lined salamander positive for Bd with low gene equivalents. All salamanders were negative after two to three treatment courses, and there were no additional mortalities. The difference in mortality and fungal load suggested that genus Aneides salamanders may be more susceptible to Bd than genus Eurycea salamanders.

Amphibians around the world are declining from threats that cannot currently be mitigated, making it impossible to safeguard some species in their natural habitats. Amphibians in the mountainous neotropics are one example where severe diseaserelated declines prompted calls for the establishment of captive assurance colonies to avoid extinctions. We surveyed experts in Panamanian amphibians to determine the probability of avoiding chytridiomycosis-related extinctions using captive breeding programs. We ranked Panamanian amphibian species by perceived susceptibility to chytridiomycosis, then calculated the likelihood of avoiding extinction as the product of three probabilities, which include (1) finding sufficient founder animals, (2) successfully breeding these species in captivity and (3) becoming extinct in the wild. The likelihood of finding enough animals to create a captive founding population was low for many rare species, especially for salamanders and caecilians. It was also low for frogs which were once regularly encountered, but have already disappeared including Atelopus chiriquiensis, Craugastor emcelae, C. obesus, C. punctariolus, C. rhyacobatrachus, Ecnomiohyla rabborum, Isthmohyla calypsa and Oophaga speciosa. Our results indicate that captive breeding could improve the odds of avoiding extinction for species that have severely declined or are likely to decline due to chytridiomycosis including Atelopus certus, A. glyphus, A. limosus, A. varius, A. zeteki, Anotheca spinosa, Gastrotheca cornuta, Agalychnis lemur and Hemiphractus fasciatus. Priority species that experts predicted were highly susceptible to chytridiomycosis that might also benefit from ex situ management include Craugastor tabasarae, C. azueroensis, C. evanesco, Strabomantis bufoniformis and Colostethus panamansis. In spite of high levels of uncertainty, this expert assessment approach allowed us to refine our priorities for captive amphibian programs in Panama and identify priority conservation actions with a clearer understanding of the probability of success.

We engaged pet salamander owners in the United States to screen their animals for two amphibian chytrid fungal pathogens Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal). We provided pet owners with a sampling kit and instructional video to swab the skin of their animals. We received 639 salamander samples from 65 species by mail, and tested them for Bd and Bsal using qPCR. We detected Bd on 1.3% of salamanders (95% CI 0.0053–0.0267) and did not detect Bsal (95% CI 0.0000–0.0071). If Bsal is present in the U.S. population of pet salamanders, it occurs at a very low prevalence. The United States Fish and Wildlife Service listed 201 species of salamanders as “injurious wildlife” under the Lacey Act (18 U.S.C. § 42) on January 28, 2016, a precautionary action to prevent the introduction of Bsal to the U.S. through the importation of salamanders. This action reduced the number of salamanders imported to the U.S. from 2015 to 2016 by 98.4%. Our results indicate that continued precautions should be taken to prevent the introduction and establishment of Bsal in the U.S., which is a hotspot of salamander biodiversity.

Batrachochytrium salamandrivorans (Bsal) is an emerging fungal pathogen that has caused recent die-offs of native salamanders in Europe and is known to be lethal to at least some North American species in laboratory trials [1]. Bsal appears to have originated in Asia, and may have been introduced by humans into wild populations in Europe through commercial trade of amphibians [1]. Since the first outbreaks of Bsal in the Netherlands, it has been the etiologic agent of mortality events in Belgium (wild) and Germany (captivity), and was recently found in imported salamanders in the United Kingdom [1–4]. Substantial concern has been raised about the associated risk of Bsal to native salamanders in North America [5]. Herein, we review what policy actions are occurring in North America and elsewhere, and call for creation of a North American Bsal Strategic Plan.