MONITORING RANAVIRUS-ASSOCIATED MORTALITY IN A DUTCH HEATHLAND IN THE AFTERMATH OF A RANAVIRUS DISEASE OUTBREAK.

The ranaviruses are an emerging group of viruses that infect amphibians, fish, and reptiles. Although ranaviruses have not been linked to extinctions, emergence in amphibian communities has resulted in population declines for some species. We present the results of ranavirus-associated mortality in a Dutch national park in the aftermath of an outbreak associated with a common midwife toad virus (CMTV)-like ranavirus. We monitored five bodies of water across Dwingelderveld National Park, the Netherlands, in 2011-13. Dead and live amphibians were counted weekly July-September and every 2 wk in June and October. Dead amphibians were collected and tested for ranavirus infection. In addition, we measured biologic, chemical, and physical site characteristics to test for a correlation with ranavirus-associated mortality. Ranavirus infection was widespread in our study area and we observed nearly continuous presence of dead, ranavirus-infected amphibians in the presence of asymptomatic, live amphibians throughout our study. Fatalities occurred in larval, subadult, and adult amphibians. Ranavirus infection prevalence (based on fatal cases) was significantly associated with increasing fractions of adults and subadults compared to juveniles and larvae in the population, but was unrelated to any other measured site characteristics. Our findings showed that a CMTV-like ranavirus can persist long term in an ecosystem, affecting a diversity of amphibian species and life stages for a prolonged period. This study illustrates the importance of monitoring the modes of spread for ranaviruses and their impact on amphibian populations.

Ranavirus-associated mass mortality in wild amphibians, the Netherlands, 2010: a first report.

In 2010, a mass die-off of over 1000 wild water frogs (Pelophylax spp.) and at least 10 common newts (Lissotriton vulgaris) occurred in a pond in The Netherlands. Haemorrhagic disease with hepatomegaly and splenomegaly was evident. Microscopically, multiple organs presented cells with multifocal intracytoplasmic inclusion bodies, in which ranavirus-like particles were demonstrated ultrastructurally. All specimens examined tested positive for ranavirus by PCR. The sequence obtained showed a 100% identity with the one deposited for common midwife toad virus (CMTV). This is the first report of ranavirus-associated mortality in wild amphibian populations in The Netherlands. It is also the first time CMTV or a CMTV-like virus has been reported in these two species in the adult stage and outside of Spain.