Species fraud and product mislabelling in processed food, albeit not being a direct health issue, often results in consumer distrust. Therefore methods for quantification of undeclared species are needed. Targeting mitochondrial DNA, e.g. CYTB gene, for species quantification is unsuitable, due to a fivefold inter-tissue variation in mtDNA content per cell resulting in either an under- (-70%) or overestimation (+160%) of species DNA contents. Here, we describe a reliable two-step droplet digital PCR (ddPCR) assay targeting the nuclear F2 gene for precise quantification of cattle, horse, and pig in processed meat products. The ddPCR assay is advantageous over qPCR showing a limit of quantification (LOQ) and detection (LOD) in different meat products of 0.01% and 0.001%, respectively. The specificity was verified in 14 different species. Hence, determining F2 in food by ddPCR can be recommended for quality assurance and control in production systems.
White Galloway cattle exhibit three different white coat colour phenotypes, that is, well marked, strongly marked and mismarked. However, mating of individuals with the preferred well or strongly marked phenotype also results in offspring with the undesired mismarked and/or even fully black coat colour. To elucidate the genetic background of the coat colour variations in White Galloway cattle, we analysed four coat colour relevant genes: mast/stem cell growth factor receptor (KIT), KIT ligand (KITLG), melanocortin 1 receptor (MC1R) and tyrosinase (TYR). Here, we show that the coat colour variations in White Galloway cattle and White Park cattle are caused by a KIT gene (chromosome 6) duplication and aberrant insertion on chromosome 29 (Cs29 ) as recently described for colour-sided Belgian Blue. Homozygous (Cs29 /Cs29 ) White Galloway cattle and White Park cattle exhibit the mismarked phenotype, whereas heterozygous (Cs29 /wt29 ) individuals are either well or strongly marked. In contrast, fully black individuals are characterised by the wild-type chromosome 29. As known for other cattle breeds, mutations in the MC1R gene determine the red colouring. Our data suggest that the white coat colour variations in White Galloway cattle and White Park cattle are caused by a dose-dependent effect based on the ploidy of aberrant insertions and inheritance of the KIT gene on chromosome 29.
The multigene family of pregnancy‐associated glycoproteins (PAGs) belongs to a group of aspartic proteases that are exclusively expressed by trophoblast cells in the placenta of even‐toed ungulates. In Bovidae, 22 different PAG genes (boPAGs) with a wide range of temporal and spatial expression‐ and glycosylation patterns have been reported to date. In this study we describe the mRNA expression patterns using real‐time quantitative PCR (qPCR) for selected modern (boPAG‐1, ‐9, ‐21) and ancient bovine PAGs (boPAG‐2, ‐8, ‐10, ‐11, ‐12) in cotyledonary tissue. The highest mean expression was detected in boPAG‐8 and lowest in boPAG‐10 (P < 0.05). Furthermore, boPAG‐8 and ‐11 were significantly greater expressed in early gestation compared with later pregnancy stages. The characterization of boPAG mRNA‐expression levels gives important insights for further protein analyses which will be valuable information for the development of new pregnancy detection systems.
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