NORTH-WESTERN JOURNAL OF ZOOLOGY 9 (2): 446-449 ©NwjZ, Oradea, Romania, 2013 Article No.: 132510 http://biozoojournals.3x.ro/nwjz/index.html First record of Batrachochytrium dendrobatidis on amphibians in Romania Judit VÖRÖS 1, *, Jaime BOSCH 2 , Ádám DÁN 3 and Tibor HARTEL 4,5,6 1. Department of Zoology, Hungarian Natural History Museum, Baross u. 13, Budapest H-1088, Hungary. 2. Museo Nacional de Ciencias Naturales, CSIC, José Gutiérrez Abascal 2, E-28006 Madrid, Spain. 3. Molecular Biology Department, Veterinary Diagnostic Directorate, National Food Chain Safety Office, 1143 Budapest, Tábornok u. 2., Hungary. 4. Faculty of Natural Sciences, Ovidius University, Constanta, Romania. 5. Sapientia University, Department of Environmental Sciences, Cluj-Napoca, Romania. 6. Institute of Ecology, Faculty of Sustainability, Leuphana University Lüneburg, Lüneburg, Germany. * Corresponding author, J. Vörös, E-mail: jvoros@nhmus.hu Received: 01. December 2012 / Accepted: 16. May 2013 / Available online: 04. June 2013 / Printed: December 2013 Abstract. Here we report for the first time the presence of Batrachochytrium dendrobatidis in Romania. Seventy- seven amphibians belonging to four species were sampled in four regions of Transylvania: Sighisoara, Corund, Miercurea-Ciuc and Apuseni Mountains. Infected individuals were found in Sighisoara and Apuseni. Long term studies in Sighisoara region suggests that no unusual mass mortality occurs in the amphibian populations sampled for this study. We suspect that amphibians coexist with the chytrid. Further studies are urgently needed to understand the origin, ways of spreading, distribution, and potential impacts of Bd on amphibian populations in this country. Key words: amphibian, chytrid, conservation, Romania. Chytridiomycosis is an emerging infectious dis- ease which is a major driver of amphibian decline in many parts of the world (e.g. Beebee & Griffiths 1995). It is caused by a fungus (Batrachochytrium dendrobatidis hereafter Bd) and it was first de- scribed on frogs from Australia and Central Amer- ica in 1998 (Berger et al. 1998). The chytrid attacks the keratinised tissues of amphibians which are the mouthpart of the larvae and the skin of the post-metamorphic individuals (Berger et al. 1998, Daszak et al. 1999). The chytrid causes severe elec- trolyte imbalance (Voyles et al. 2006) and affects respiration ultimately resulting in the death of the individuals (Berger et al. 1998). Mass mortality due to Bd was reported for the adult stages as well as metamorphs (e.g. Bosch et al. 2001), and lower survival for larval stages (e.g. Garner et al. 2009). Having limited ability to swim, the fungus is likely carried in long distances by human activi- ties, the animals or water flow (Johnson & Speare 2005). The first record of Bd in the wild in European amphibians is from 1997 from Peňalara Mountains (Bosch et al. 2001). The occurrence of Bd in Europe was further reported from Portugal, Italy, France, Germany, Switzerland, Denmark, UK, Austria, Czech Republic, Poland and Hungary (Garner et al. 2005, Federici et al. 2008, Scalera et al. 2008, Sura et al. 2010, Ohst et al. 2011, Sztatecsny & Glaser 2011, Civis et al. 2012, http://www.bd- maps.net/). Currently the Easternmost part of Europe where Bd was confirmed is Hungary (Gál et al. 2012). Here we report for the first time the pres- ence of Bd in two regions of Romania. Amphibians were sampled in four regions of Transylva- nia (Romania) (Fig. 1) the Apuseni mountains (hereafter Apuseni, with 26 individuals belonging to Bombina varie- gata and Lissotriton vulgaris ampelensis), Sighisoara area (41 individuals belonging to two species), Corund area (here- after Corund, with one specimen belonging to B. varie- gata) and Miercurea Ciuc area (hereafter Ciuc, with 11 in- dividuals belonging to Rana temporaria, R. arvalis and L. vulgaris ampelensis). Two sites were sampled in Sighisoara: the Breite wood-pasture (at an altitude of ca 430 m) and a deciduous forest (at ca 480 m altitude). In Apuseni six lo- cations were sampled; two of them (Nucet, Baita) at an al- titude of between 400-550 m, two sites at ca 800 m (Mununá, Valea largá) and two (Vartop, Padis) at ca 1100 m altitude. The site from Corund was at 650 m altitude and the site Ciuc at 700 m altitude (Table 1). Data were collected in April 2011 (for Sighisoara, Co- rund and Ciuc) and in July 2011 (Apuseni). We used sterile dry swabs (Biolab and Medical Wire and Equipment) to sample the back, belly, sides and legs of the specimens. The samples were then transferred to Hungary and were processed in the Laboratory for Mo- lecular Taxonomy of the Hungarian Natural History Mu- seum and the Veterinary Diagnostic Directorate of the National Food Chain Safety Office. To detect Bd we used the real-time PCR protocol by Boyle et al. (2004). We ran every sample in duplicate with a dedicated internal posi- tive control (TaqMan Exogenous Internal Positive Control Reagents) for each sample that helps to identify inhibitors present in the DNA extractions. The templates were run