SHORT COMMUNICATION Low neutral genetic variability in a specialist puffin hunter: the Norwegian Lundehund Claudia Melis*, A ˚ sa A. Borg*, Ingvild S. Espelien † and Henrik Jensen* *Centre for Conservation Biology, Department of Biology, Norwegian University of Science and Technology, 7491, Trondheim, Norway. † Norwegian Lundehund Club, Breeding Council, Tangen, 7039, Trondheim, Norway. Summary The genetic variability of 125 Norwegian Lundehund and 27 Nova Scotia Duck Tolling Retriever was analysed using a set of 26 microsatellite markers. In Lundehund, the average number of alleles per locus was 1.73, and average observed (H O ) and expected (H E ) heterozygosity were 0.07. In Toller, all measures of genetic diversity were much higher than in Lundehund and similar to studies on other dog breeds. The cluster analysis correctly assigned individuals to their respective breed. The low genetic variability in Lundehund was not surprising, given the two strong bottlenecks in the 1940s and the 1960s. The relatedness of Lundehund to other Nordic small spitzes should be investigated in the view of possible outcrossing. Keywords Canis familiaris, genetic bottleneck, inbreeding, Nova Scotia Duck Tolling Retriever, puffin dog The Norwegian Lundehund is a small spitz, which since the 17th century was used to hunt puffins (Fratercula arctica, lunde in Norwegian) on steep mountainsides along the coast of northern Norway. This breed is characterized by a great flexibility of the joints and by polydactyly (i.e. the presence of extra toes). At the beginning of the 20th century, only a small population of Lundehund was left in the fishing village of Ma ˚stad (Værøy island, Lofoten archipelago). The breed went through two bottlenecks, one caused by canine distemper in the 1940s and one following the abandonment of Ma ˚stad in the 1960s. Today’s Lundehund world popu- lation stems entirely from five surviving dogs, who shared a grandmother. Three of them also shared the mother (Frimann-Clausen & Laane 1968). A recent census (2010, Norwegian Lundehund Club) found that there are ca. 500 Lundehund in Norway and ca. 300 in other European countries. The aim of this study was to evaluate Lundehund within breed genetic diversity; the same analyses were performed on a sample of Nova Scotia Duck Tolling Retriever (hereafter Toller) for comparison. A total of 125 Lundehund and 27 Toller were DNA sampled by buccal swabs at dog shows and meetings in Norway (2009–2011). DNA was extracted using the Isohelix DDK-50 isolation kit. A total of 22 microsatellite markers from the ISAG panel (http://www.isag.us/Docs/consignmentforms/2005ISAG PanelDOG.pdf) and nine extra microsatellites (http://www. qiagen.com/literature/qiagennews/weeklyarticle/09_06/e10/ default.aspx) were chosen to perform the molecular anal- yses. Amelogenin was used for sex identification. After exclusion of four markers that presented problems in amplification or scoring, 26 autosomal markers remained and were amplified by PCR in three multiplex panels (Table 1). All individuals were genotyped at the 27 loci in 10-ll reactions as described in Kekkonen et al. (2011), and alleles were scored using GENEMAPPER 4.0 software (Applied Biosystems). For each locus, observed (H O ) and expected (H E ) hetero- zygosity and number of alleles were calculated by CERVUS 3.0.3 (Kalinowski et al. 2007). The inbreeding coefficient (F IS ) and allelic richness corrected for sample size (A R ) were calculated in FSTAT 2.9.3.2 (Goudet 1995). Within each breed, we tested for locus-specific departure from Hardy– Weinberg (HW) equilibrium using exact tests in GENEPOP 3.4 (Raymond & Rousset 1995). All markers were in HW equilibrium after Bonferroni correction except C01.246 in Lundehund (Table 1). We also tested for significant linkage disequilibrium (LD) with GENEPOP 3.4. In Lundehund, only 34 of 325 combinations could be tested owing to the presence of many monomorphic loci and very few alleles on variable loci, but significant LD was found in 1.84% of pairs Address for correspondence C. Melis, Centre for Conservation Biology, Department of Biology, Norwegian University of Science and Technology, Trondheim 7491, Norway. E-mail: claudia.melis@bio.ntnu.no Accepted for publication 07 August 2012 doi: 10.1111/age.12000 1 © 2012 The Authors, Animal Genetics © 2012 Stichting International Foundation for Animal Genetics