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Dear all, In the Netherlands different institutions raise toads from the common spade foot toad or Garlic Toad (Pelobates fuscus) for reintroduction purposes. The eggs were collected in many different parts of the country. At this moment the larvae are supposed to be released in nature. But Ravon (reptile, amphibian and fish research Netherlands) wants the animals tested on chytrid an ranavirus before releasing them. Swabs are being examined for chytrid infection by PCR. But the main challenge now is how to detect in a reliable way whether ranavirus is present within these groups of animals. The following questions came from the involved veterinarian. How many animals should be tested to have a statistical reliability that they are really free of ranavirus infection. The animals are housed in separated tanks. In some of them approximately 100 are housed, in others a thousand. Further what would you use as test samples? Is a piece of tail? The whole animal? I'm aware if the work of Greer and Collins and the risk of underestimating the true prevalence of infection, as PCR does for early-stage infections. But please could you advise us how to proceed. All the best Marja