Growth and development traits are economically important in animal production, especially in pig breeding. Therefore, the porcine GHRL gene is considered as a candidate gene responsible for growth rate and body weight. The aim of our study was to identify new polymorphisms in the GHRL gene in pig. Ten novel single nucleotide polymorphisms (SNP's) were detected: four substitutions in exons, four in introns and two mutations in promoter region. We evaluated the GHRL mRNA abundance in porcine stomachs (fundus ventriculis) and ghrelin protein concentration in plasma in three breeds: Polish Landrace, Polish Large White and Pietrain. The results showed that transcript abundance of GHRL gene was significantly higher in Polish Landrace than in other two breeds (P<0.05). The mutation c.-93A>G located in the promoter region affected expression of the GHRL gene. The AA genotype animals showed a significantly (P<0.05) higher expression level when compared to AG genotype animals.
NATONEK-WINIEWSKA M., S£OTA E., KALISZ B. 2010. Use of cytochrome b polymorphism for species identification of biological material derived from cattle, sheep, goats, roe deer and red deer. Folia biol. (Kraków) #&: 47-50. The objective of this study was species identification of the following biological trace material: skin, blood stains, meat samples and jawbone with a tooth, which were the subject of expert opinion ordered by a court. The expert appraisement was conducted by an analysis of a cytochrome b fragment. The choice of mtDNA fragment for analysis was based on its conservation in mammals which enabled several farm and wild species to be identified with one pair of primers. The PCR product was differentiated by Tsp509I and AluI enzymes. Due to problems with amplification of roe deer DNA, primers specific to this species only, flanking a cytochrome b fragment (Y14951.1), were designed. On the basis of this analysis, it was concluded that the skin sample was derived from a goat, dried blood from a roe deer, the jawbone from cattle, and two meat samples from a roe deer and red deer. This method allowed rapid and efficient identification of several species of mammals using diverse biological material.
This preliminary study aimed to differentiate domestic pigs from wild boars based on MC1R and NR6A1 polymorphisms and to identify admixture between these genomes. We studied samples obtained from wild boars from two regions of Poland and five pig breeds: Polish Landrace, Polish Large White, Złotnicka White, Pulawska and Duroc. Along the MC1R gene sequence, we identified four polymorphic loci comprising three codons. The “wild type” allele was primarily found in wild boar but also in the Duroc and Złotnicka White breeds. Non-wild type alleles were identified in the vast majority of domestic pig samples and in two wild boar samples. Based on MC1R profiles, we conducted a population study, and revealed admixture between both genomes using STRUCTURE and NETWORK Software. Interestingly, an allelic discrimination assay with NR6A1 g.748C > T TaqMan probes revealed a clear separation of samples into two groups: wild boar samples representing the C allele and domestic breeds representing the T allele. Based on the obtained results, we conclude that NR6A1 g.748C > T is an effective marker for differentiating between wild boars and domestic pigs, where this is supported by MC1R data, to identify admixed profiles. We recommend that a larger sample of genomes is studied to verify this method.
european union regulations (e.g. Commission Regulation (eu) no 56/2013) set restrictions on the use of poultry meals in animal nutrition, which requires the species composition of manufactured feeds to be constantly monitored. The aim of this study is to develop a method for qualitative analysis of poultry meals, enabling the four main poultry species (chicken, turkey, duck, goose) to be detected using one pair of primers, and a method for quantitative determination of chicken meals in poultry meals. The qualitative identification method was developed using PCR technology, whereas qPCR and TaqMan probes were used for quantitative identification. The study was performed with samples of feed mixture containing poultry and chicken meal. The limit of determination was 0.08% and 0.02% for qualitative and quantitative identification, respectively. The results of quantitative determinations obtained for independent dna isolations from the same samples are repeatable (RSDcT ≤0.46%). The determined concentrations are accurate (Dc ≤11.23% for c ≥0.06). The identification of target sequences in both tests is good enough for commercial applications.
The objective of the study was to demonstrate how mitochondrial DNA (mtDNA) can be used to determine the species origin of animal microtraces. The study included pieces of cat and dog hair without the root, a fragment of cooked chicken bone (0.1g), three goose down samples (0.028 g), a pork swab, a pork scratching (5×5×5 mm), and pork lard (0.22 g). DNA was isolated from all of these samples using the method appropriate for the particular source material. The extracts had DNA concentration exceeding 5.4 ng/µl with A purity range of 1.14-1.88. Next, the samples were subjected to PCR and real-time PCR with species-specific primers and primers complementary to mitochondrial DNA (mtDNA). Control reactions based on the amplification of eukaryotic-specific fragment (18S rRNA) were additionally performed. PCR and real-time PCR products for detection of species-specific mtDNA were obtained for all templates, whereas during the detection of eukaryote DNA no product was obtained for dog and cat hair only. The poor quality of the obtained DNA did not prevent the analysis. The results showed that mitochondrial DNA is suitable for identification of small or highly processed samples, in which genomic DNA often cannot be analyzed.
Gilts were slaughtered at 60 th , 90 th , 120 th , 150 th , 180 th and 210 th day of breeding (6 sows in each age). After afeing in 4 0 C during 24 hours the samples were taken from right half of carcasses of longissimus dorsi (behind the last rib between the thoracic and lumbar vertebrae) muscle. In the meat to carry out instrumental measurement of shear force and texture parameters. The meat of loin was subjected to chemical analysis to determine its dry matter, water, protein, fat and ash content. The age of fatteners slaugthering beside the genotype and sex is the basic factor influencing on quality, nutritional value and attractiveness for consumer of pig meat.
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