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OUELLET, M., I. MIKAELIAN, B. D. PAULI, J. RODRIGUE, AND D. M. GREEN.2005. Historical evidence of widespread chytrid infection in NorthAmerican amphibian populations. Conserv. Biol. 19:1431–1440.PIOTROWSKI, J. S., S. L. ANNIS, AND J. E. LONGCORE. 2004. Physiology ofBd, a chytrid pathogen of amphibians. Mycologia 96:9–15.REGOSIN, J. V., B. S. WINDMILLER, AND J. M. REED. 2003. Terrestrial habitatuse and winter densities of the wood frog (Rana sylvatica). J. Herpetol.37:390–394.RELYEA, R. A. 2005. The lethal impact of roundup on aquatic and terrestrialamphibians. Ecol. Appl. 15:1118–1124.RETALLICK, R. W. R., H. MCCALLUM, AND R. SPEARE. 2004. Endemic infectionof the amphibian chytrid fungus in a frog community post-decline.Plos Biol. 2:1965–1971.RON, S. R. 2005. Predicting the distribution of the amphibian pathogenBatrachochytrium dendrobatidis in the New World. Biotropica 37:209–221.SANZO, D., AND S. J. HECNAR. 2006. Effects of road de-icing salt (NaCl) onlarval wood frogs (Rana sylvatica). Environ. Pollut. 140:247–256.SKERRATT, L. F., L. BERGER, R. SPEARE, S. CASHINS, K. R. MCDONALD, A.D. PHILLOTT, H. B. HINES, AND N. KENYON. 2007. Spread ofchytridiomycosis has caused the rapid global decline and extinction offrogs. EcoHealth 4:125–134.WOODHAMS, D. C., AND R. A. ALFORD. 2005. Ecology of chytridiomycosisin rainforest stream frog assemblages of tropical Queensland. Conserv.Biol. 19:1449–1459.––––––, ––––––, AND G. MARANTELLI. 2003. Emerging disease of amphibianscured by elevated body temperature. Dis. Aquat. Org. 55:65–67.Herpetological Review, 2008, 39(2), 199–200.© 2008 by Society for the Study of Amphibians and ReptilesOccurrence of Batrachochytrium dendrobatidis inAmphibian Populations in Denmark(pers. comm. to R. Scalera, 2007). Here, we report the results ofsurveys carried out at four sites in Denmark (Fig. 1) on two nativeamphibians: Rana temporaria and Rana kl. esculenta.In summer 2007, we hand captured individual amphibians andsampled them for B. dendrobatidis by rubbing a cotton-tipped swabover the body of each individual. Frogs were held separately priorto swabbing and technicians wore a new pair of gloves for eachindividual handled. The sampling is harmless and was carried outin-situ so as to release the sampled animals within just a few minutesat the location where they were captured. As the frog wasrestrained, the swab was firmly rubbed back and forth 25–30 times,targeting the drink patch, the mouth, and the webbing betweeneach toe. The swab was immediately inserted, cotton side down,into a 2 ml screw-cap tube containing 1 ml of 70% ethanol andstored upright. Vials were shipped to the laboratory for analysis,and each swab was analyzed individually for the presence of B.dendrobatidis. Swabs were qualitatively analyzed using a PCRassay (45 amplification cycles). Presence of B. dendrobatidis wasdetermined by presence of PCR product visualized on agarose gels(30–90 minute electrophoresis) containing positive controls. Fragmentswere sized using a molecular weight marker (Pisces MolecularLLC, Boulder, Colorado, USA (Annis et al. 2004; J. Wood,pers. comm.). All field gear was cleaned with a brush and waterand then sterilized using a dilute bleach solution between eachsampling location.Two of the 13 amphibians we swabbed were positive for B.dendrobatidis (Table 1). We found B. dendrobatidis on individualsfrom both species and at 2 of the 4 study areas we examined.One of the positive results was for an adult of Rana kl. esculentacaptured in Vestamager. The other positive result was for a juvenileof Rana temporaria captured in Egense. We did not find anyfrogs that were dead or that appeared to be sick.RICCARDO SCALERA*Via Torcegno 49 V1 A2, Rome 00124, ItalyMICHAEL J. ADAMSandSTEPHANIE K. GALVANU. S. Geological Survey, Forest and Rangeland Ecosystem Science Center3200 SW Jefferson Way, Corvallis, Oregon 97331, USA*Corresponding author: Riccardo.Scalera@alice.itAmphibian decline is a global phenomenon with multiple causes(Stuart et al. 2004). Some declines have been attributed to thedisease chytridiomycosis that affects the skin of amphibians(Skerratt et al. 2007). The agent responsible for chytridiomycosisis the fungus Batrachochytrium dendrobatidis (Berger et al. 1998).There is evidence that the spread of B. dendrobatidis around theworld occurred in the last half century (Ouellet et al. 2005), andthere is a need for detailed information on its current spatial extent.In Europe, B. dendrobatidis has been reported in several amphibianspecies in multiple countries, such as Spain, Portugal, Italy,Switzerland, France, Germany and the UK (Cunningham et al.2005; Garner et al. 2005, 2006; Mutschmann et al. 2000; Simoncelliet al. 2005; Stagni et al. 2004). No comprehensive surveys haveoccurred in Denmark but a single record of B. dendrobatidis forRana kl. esculenta on the island of Bornholm is reported(www.spatialepidemiology.net) and confirmed by Trent GarnerFIG. 1. Locations of study areas in Denmark where amphibians weresampled for the presence of Batrachochytrium dendrobatidis in 2007.Circles are filled at locations where we found B. dendrobatidis. The squaresymbol indicates the location of the positive record reported by TrentGarner (see text). Vestamager is located on the island of Zealand, close toCopenhagen, Egense is on Fyn Island, and both Amtoft and Klosterhedenare on the Jutland Peninsula.Herpetological Review 39(2), 2008 199