<i>COL7A1</i> genotype frequencies.

*<p>Angus (n = 18), Aubrac (n = 1), Ayshire (n = 1), Belgian blue (n = 3), Blonde d'Aquitaine (n = 2), Brown Swiss (n = 35), Charolais (n = 17), Chianina (n = 19), Dutch belted (n = 18), Eringer (n = 16), Evolenard (n = 10), Gelbvieh (n = 1), Galloway (n = 20), Hereford (n = 3), Scotish Highland (n = 4), Holstein (n = 35), Jersey (n = 3), Limousin (n = 17), Montbéliarde (n = 4), Nelore (n = 3), Ongola (n = 1), Piedmontese (n = 2), Pinzgauer (n = 10), Pustertaler Sprinzen (n = 10), Romagnola (n = 18), Salers (n = 1), Simmentaler (n = 38), Tyrolean Grey (n = 18), Domestic Yak (n = 1).</p

Pedigree of Rotes Höhenvieh cattle with DEB.

<p>DNA samples were available only from red labeled cattle. The genotypes for the <i>COL7A1</i> c.4756C>T exon 49 SNP are given below the symbols. The arrow indicates the cow, which is supposed to be the founder animal. We genotyped all available male offspring of the two carrier bulls Hannibal and Oska and detected 51% and 61% carriers, respectively.</p

Genome-wide homozygosity mapping of the DEB mutation.

<p>After genotyping approximately 777,000 uniformly distributed SNP markers homozygous blocks >0.1 Mb were identified across 3 DEB affected cattle. Only on BTA 22 a very large homozygous block was identified.</p

Clinical features of a DEB affected Rotes Höhenvieh calf.

<p>Typical signs are the extensive epidermal loss with ulcerations at the fetlocks (A) and muzzle (B) and dysungulation (C).</p

<i>COL7A1</i> mutation analysis.

<p>Electropherograms of the <i>COL7A1</i> c.4756C>T mutation. Representative sequence traces of PCR products amplified from genomic DNA of 3 cattle with the different genotypes are shown. The mutant T allele in homozygous form is present only in DEB affected calves and leads to a premature stop codon.</p

Histopathological features of DEB.

<p>Biopsy of the ear of case 1: (A) Note that on the lateral borders of the biopsy the epidermis (ep) is missing and, where present, the epidermis is detached from the underlying dermis (de). The epidermis is cleanly separated from the dermis and the basal cells are intact. H&E, 25×. (B) Higher magnification: Note the intact basal cells of the blister roof (br), larger vacuoles (lv) and a small vesicle (sv) along the basement membrane zone and separation of the infundibular epithelium (ie) from the surrounding connective tissue. H&E, 200× (C) Note that the epidermal and infundibular epithelium of the hair follicle (hf) is missing and the surface is covered by homogenous proteinaceous material. H&E, 200×. Biopsy of the right hind leg of case 3: (D) Necrotic superficial dermis (de) characterized by fibrinous exsudation (fe), cellular debris, dense amounts of degenerate inflammatory cells, and hemorrhage. H&E, 100×.</p

Multispecies alignment of the region around the KDM2B:p.D835N mutation.

<p>Multispecies alignment of the region around the KDM2B:p.D835N mutation.</p

Genome-wide association and homozygosity mapping of the paunch calf syndrome locus.

<p>(<b>A</b>) Case-control genome-wide association analysis shows a significant association to SNPs on BTA 17. (<b>B</b>) Single SNP association results across BTA 17. (<b>C</b>) The limit of the homozygous 1.23 Mb interval shared by the 16 cases is highlighted in red. (<b>D</b>) Gene content of the corresponding human chromosome 5 segments. The <i>KDM2B</i> gene containing the causative mutation is highlighted in red.</p

Pedigree of selected inbred Romagnola cattle.

<p>The special structure of livestock families facilitates the mutation identification: The sire A, born 1969, is the assumed founder animal for paunch calf syndrome. The paunch calf syndrome mutation (indicated by an asterisk) most likely occurred in the germline of sire A. Beneath the animals the two copies of BTA 17 are indicated. Black symbols represent the ancestral chromosome, on which the mutation occurred. Gray symbols represent any other wild-type copy of BTA 17. Due to inbreeding loops three animals (cow 326, cow 19, bull 145) have inherited two copies of the same chromosome from their common ancestor (sire A). These three animals were genotyped as heterozygous carriers of the <i>KDM2B</i> mutation. For the cow 19 it is shown that her paternal copy of the critical 1.23 Mb interval is still in its ancestral wild-type state as her father (bull 198) was tested <i>KDM2B</i> clear, while her maternal copy carries the paunch calf syndrome mutation. The BTA 17 haplotypes were confirmed by SNP genotyping of available samples from cow 326, cow 19, bull 198 and bull 145. Based on pedigree and marker data these three animals were identical-by-descent for the critical segment on BTA 17, with the only exception of the causative paunch calf syndrome mutation.</p

Genotyping results for <i>RNF34</i> and <i>KDM2B</i> mutations.

a<p>Cattle recorded as parent of affected calves.</p>b<p>For this cow only one single suspicious offspring was recorded by the breeding organization and the diagnosis of this calf had not been confirmed by necropsy. Genotyping of additional flanking markers revealed that this animal did not carry the disease-associated haplotype. Therefore, the reported calf most likely represented a phenocopy and the parent is indeed free of the deleterious mutation.</p