Investigation of Amphibian Mortality Events in Wildlife Reveals an On-Going Ranavirus Epidemic in the North of the Netherlands.

In the four years following the first detection of ranavirus (genus Ranavirus, family Iridoviridae) infection in Dutch wildlife in 2010, amphibian mortality events were investigated nationwide to detect, characterize and map ranaviruses in amphibians over time, and to establish the affected host species and the clinico-pathological presentation of the disease in these hosts. The ultimate goal was to obtain more insight into ranavirus disease emergence and ecological risk. In total 155 dead amphibians from 52 sites were submitted between 2011 and 2014, and examined using histopathology, immunohistochemistry, virus isolation and molecular genetic characterization. Ranavirus-associated amphibian mortality events occurred at 18 sites (35%), initially only in proximity of the 2010 index site. Specimens belonging to approximately half of the native amphibian species were infected, including the threatened Pelobates fuscus (spadefoot toad). Clustered massive outbreaks involving dead adult specimens and ranavirus genomic identity indicated that one common midwife toad virus (CMTV)-like ranavirus strain is emerging in provinces in the north of the Netherlands. Modelling based on the spatiotemporal pattern of spread showed a high probability that this emerging virus will continue to be detected at new sites (the discrete reproductive power of this outbreak is 0.35). Phylogenetically distinct CMTV-like ranaviruses were found in the south of the Netherlands more recently. In addition to showing that CMTV-like ranaviruses threaten wild amphibian populations not only in Spain but also in the Netherlands, the current spread and risk of establishment reiterate that understanding the underlying causes of CMTV-like ranavirus emergence requires international attention.

Concurrent ranavirus and Batrachochytrium dendrobatidis infection in captive frogs (Phyllobates and Dendrobates species), The Netherlands, 2012: a first report.

A ranavirus infection with concurrent Batrachochytrium dendrobatidis infection and mortality in captive Phyllobates and Dendrobates species is reported. Greyish skin with hepato- and reno-megaly were evident. Microscopically, Batrachochytrium dendrobatidis was present in the stratum corneum of the hyperkeratotic skin. Intracytoplasmic inclusion bodies were present in erythrocytes and multiple organs. All samples examined tested positive using PCR for the major capsid protein (MCP) gene of ranavirus and the ITS-1-5.8S region of B. dendrobatidis. The sequence obtained showed a 99% identity with the deposited sequence of the MCP gene of the common midwife toad virus (CMTV). This is the first report of mortality in captivity in poison dart frogs caused by a ranavirus, CMTV or like virus, and Batrachochytrium dendrobatidis infection.