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Hi Rolando and others 1. I am afraid not. 2. Yes we had ranavirus in the Netherlands see attachment. 3. Very small, I asked the involved vet, but he told me he swabbed the whole animal also the mouthparts??? I agree completely with your other comment. There is a firm that is trying to develop environmental DNA tests for ranavirus, but is not ready with it yet. Thanks for all the input!! Gr Marja Van: Global Ranavirus Consortium [mailto:[log in to unmask]] Namens Rolando Mazzoni Verzonden: woensdag 20 juni 2012 1:44 Aan: [log in to unmask] Onderwerp: Re: question from the netherlands Hi Marja and all colleagues, Previous emails have given exellent information about sampling from which I will take great advantage. So, many thanks ! Concerning the initial question I think it will be useful to know some details for risk assessment considerations: 1. The larvae will be released in the same area where adults were caught ? 2. Do you have any previous diagnostic/detection of Bd or ranavirus in your country or surrounding areas? 3. Which size are the tadpoles? Because you said that Bd detection is based on swabs, but the parasite will be present only in keratinized mouth tissues, probably difficult to be sampled by this method. In addition, haematopoietic tissues targeted for ranavirus detection will be hard to get in very small tadpoles, where we have succeed using the whole body for DNA extraction. I hope these questions will help to the discussion. Best regards, Rolando Mazzoni DVM/PhD CPA/EV/UFG Brazil On Mon, Jun 18, 2012 at 7:13 AM, marja kik <[log in to unmask]> wrote: 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