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Hi Greg thanks I mailed him, and have answers back from others Gr Marja Van: Global Ranavirus Consortium [mailto:[log in to unmask]] Namens Victor Gregory Chinchar Verzonden: maandag 18 juni 2012 16:09 Aan: [log in to unmask] Onderwerp: Re: question from the netherlands The fellow to ask is Alex Hyatt as he has worked up the sampling protocols for amphibian ranaviruses. Protocols are in the OIE Manual, but I'd drop Alex an email. His email address is (and I suspect he is part of the GRC): [log in to unmask] Greg From: Global Ranavirus Consortium [mailto:[log in to unmask]] On Behalf Of marja kik Sent: Monday, June 18, 2012 5:13 AM To: [log in to unmask] Subject: question from the netherlands 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 Individuals who have received this information in error or are not authorized to receive it must promptly return or dispose of the information and notify the sender. Those individuals are hereby notified that they are strictly prohibited from reviewing, forwarding, printing, copying, distributing or using this information in any way.