Temperature and duration of exposure drive infection intensity with the amphibian pathogen Batrachochytrium dendrobatidis.

The intensity of a pathogen infection plays a key role in determining how the host responds to infection. Hosts with high infections are more likely to transmit infection to others, and are may be more likely to experience progression from infection to disease symptoms, to being physiologically compromised by disease. Understanding how and why hosts exhibit variation in infection intensity therefore plays a major part in developing and implementing measures aimed at controlling infection spread, its effects, and its chance of persisting and circulating within a population of hosts. To track the relative importance of a number of variables in determining the level of infection intensity, we ran field-surveys at two breeding sites over a 12 month period using marked larvae of the common midwife toad (Alyes obstetricans) and their levels of infection with the amphibian pathogen Batrachochytrium dendrobatidis (Bd). At each sampling occasion we measured the density of larvae, the temperature of the water in the 48 h prior to sampling, the period of time the sampled individual had been in the water body, the developmental (Gosner) stage and the intensity of Bd infection of the individual. Overall our data suggest that the temperature and the duration of time spent in the water play a major role in determining the intensity of Bd infection within an individual host. However, although the duration of time spent in the water was clearly associated with infection intensity, the relationship was negative: larvae that had spent less than 3-6 months in the water had significantly higher infection intensities than those that had spent over 12 months, although this infection intensity peaked between 9 and 12 months. This could be due to animals with heavier infections developing more quickly, suffering increased mortality or, more likely, losing their mouthparts (the only part of anuran larvae that can be infected with Bd). Overall, our results identify drivers of infection intensity, and potentially transmissibility and spread, and we attribute these differences to both host and pathogen biology.

Host contribution to parasite persistence is consistent between parasites and over time, but varies spatially.

Most parasites and pathogens infect multiple hosts, but a great deal of variation exists in the role of those hosts in persistence of infection. Understanding which hosts are most important in maintaining parasites can provide a clearer target for infection control. Recently developed empirical and theoretical approaches provide a way to quantify the relative contribution of hosts within a community and place them in a multi-host framework to better direct control efforts. Amphibians provide a framework for better understanding multi-host-multi-parasite dynamics. Two well-studied amphibian parasites, Batrachochytrium dendrobatidis (Bd) and Ranavirus, infect multiple host species and exhibit a great deal of heterogeneity in how they affect hosts. We used these two parasites and a community of five amphibian species to investigate the relative importance of hosts in parasite persistence, and how any patterns varied spatially and temporally. At two sites (Lake Ercina and Lake Lloroza in the Picos de Europa National Park, Spain) we collected data on the prevalence and shedding rate of parasite infection for both Bd and Ranavirus, and the abundance of each species' life stages. We used these data to parameterize a recently developed modeling framework, which was used to quantify the relative contribution of each host to the community reproductive number, R0 . By comparing each host-category over time and between sites we were able to identify consistencies in which host was responsible for the maintenance of these two parasites. Within a site one species consistently contributed the most to the persistence of both parasites. This consistency did not transfer between sites, the maintenance host species being different for each. At one site (Ercina), life stages of the common midwife toad, Alytes obstetricans, acted as the maintenance host for both Bd and Ranavirus. In contrast, at the second site, Lloroza, the alpine newt, Ichthyosaura alpestris, fulfilled that role. A single host species was responsible for infection persistence of both parasites at each lake. Attempts to control the infection levels and impacts of multiple parasites can benefit from a community epidemiology approach, and provide clarity on which hosts are the foci of mitigation efforts. However, at a small spatial scale, the target host may vary according to the physical qualities of those sites and the demographics of the host community.

Single infection with Batrachochytrium dendrobatidis or Ranavirus does not increase probability of co-infection in a montane community of amphibians.

Understanding the occurrence and consequence of co-infections can be useful in designing disease management interventions. Amphibians are the most highly threatened vertebrates, and emerging pathogens are a serious threat to their conservation. The amphibian chytrid fungus and the viruses of the Ranavirus genus are already widely distributed, causing disease outbreaks and population declines worldwide. However, we lack information about the occurrence and consequences of coinfection with these pathogens across age-classes of amphibian hosts. Here, we analyze the occurrence of infection of the amphibian chytrid fungus and ranaviruses during one season in two susceptible amphibian species at two different locations at which outbreaks have occurred. We found that the co-occurrence of both pathogens in a particular host is not common except in highly susceptible life-stages, and that single infections are the most common situation. Moreover, we found that the occurrence of one pathogen in a particular host did not predict the occurrence of the other. We attribute these results to the niches in which both pathogens proliferate in amphibian hosts.

Seasonal migrations, body temperature fluctuations, and infection dynamics in adult amphibians.

Risks of parasitism vary over time, with infection prevalence often fluctuating with seasonal changes in the annual cycle. Identifying the biological mechanisms underlying seasonality in infection can enable better prediction and prevention of future infection peaks. Obtaining longitudinal data on individual infections and traits across seasons throughout the annual cycle is perhaps the most effective means of achieving this aim, yet few studies have obtained such information for wildlife. Here, we tracked spiny common toads (Bufo spinosus) within and across annual cycles to assess seasonal variation in movement, body temperatures and infection from the fungal parasite, Batrachochytrium dendrobatidis (Bd). Across annual cycles, toads did not consistently sustain infections but instead gained and lost infections from year to year. Radio-tracking showed that infected toads lose infections during post-breeding migrations, and no toads contracted infection following migration, which may be one explanation for the inter-annual variability in Bd infections. We also found pronounced seasonal variation in toad body temperatures. Body temperatures approached 0 °C during winter hibernation but remained largely within the thermal tolerance range of Bd. These findings provide direct documentation of migratory recovery (i.e., loss of infection during migration) and escape in a wild population. The body temperature reductions that we observed during hibernation warrant further consideration into the role that this period plays in seasonal Bd dynamics.