In vitro thermal tolerance of a hypervirulent lineage of Batrachochytrium dendrobatidis: Growth arrestment by elevated temperature and recovery following thermal treatment.

Chytridiomycosis, an emerging infectious disease caused by Batrachochytrium dendrobatidis (Bd), poses a serious threat to amphibians. The thermal optimum of Bd is lower than that of most amphibians, providing an opportunity to cure infected individuals with elevated temperature. However, this approach presupposes detailed knowledge about the thermal tolerance of the fungus. To determine the temperature that may effectively reduce infection burdens in vivo, detailed in vitro studies are needed to characterize thermal tolerance of the fungus without complexities introduced by the species-specific characteristics of hosts' immune systems. The aim of our study was to evaluate the thermal tolerance of a hypervirulent isolate of Bd, considering the limits of its thermal tolerance and its capacity to rebound following heat treatment. We incubated Bd cell cultures at five different temperatures (21, 25.5, 27, 29, or 30.5 C) for one of six exposure durations (3, 4, 5, 6, 7, or 8 days) and subsequently counted the number of zoospores to assess the temperature dependence of Bd growth. We observed intensive Bd growth at 21 C. At 25.5 C, the number of zoospores also increased over time, but the curve plateaued at about half of the maximum values observed in the lower temperature treatment. At temperatures of 27 C and above, the fungus showed no measurable growth. However, when we moved the cultures back to 21 C after the elevated temperature treatments, we observed recovery of Bd growth in all cultures previously treated at 27 C. At 29 C, a treatment duration of 8 days was necessary to prevent recovery of Bd growth, and at 30.5 C a treatment duration of 5 days was needed to achieve the same result, revealing that these moderately elevated temperatures applied for only a few days have merely a fungistatic rather than a fungicidal effect under in vitro conditions.

Host-multiparasite interactions in amphibians: a review.

Parasites, including viruses, bacteria, fungi, protists, helminths, and arthropods, are ubiquitous in the animal kingdom. Consequently, hosts are frequently infected with more than one parasite species simultaneously. The assessment of such co-infections is of fundamental importance for disease ecology, but relevant studies involving non-domesticated animals have remained scarce. Many amphibians are in decline, and they generally have a highly diverse parasitic fauna. Here we review the literature reporting on field surveys, veterinary case studies, and laboratory experiments on co-infections in amphibians, and we summarize what is known about within-host interactions among parasites, which environmental and intrinsic factors influence the outcomes of these interactions, and what effects co-infections have on hosts. The available literature is piecemeal, and patterns are highly diverse, so that identifying general trends that would fit most host-multiparasite systems in amphibians is difficult. Several examples of additive, antagonistic, neutral, and synergistic effects among different parasites are known, but whether members of some higher taxa usually outcompete and override the effects of others remains unclear. The arrival order of different parasites and the time lag between exposures appear in many cases to fundamentally shape competition and disease progression. The first parasite to arrive can gain a marked reproductive advantage or induce cross-reaction immunity, but by disrupting the skin and associated defences (i.e., skin secretions, skin microbiome) and by immunosuppression, it can also pave the way for subsequent infections. Although there are exceptions, detrimental effects to the host are generally aggravated with increasing numbers of co-infecting parasite species. Finally, because amphibians are ectothermic animals, temperature appears to be the most critical environmental factor that affects co-infections, partly via its influence on amphibian immune function, partly due to its direct effect on the survival and growth of parasites. Besides their importance for our understanding of ecological patterns and processes, detailed knowledge about co-infections is also crucial for the design and implementation of effective wildlife disease management, so that studies concentrating on the identified gaps in our understanding represent rewarding research avenues.