Single Nucleotide Polymorphisms in collagen genes and association with skin quality trait

AbstractLivestock skin is largely employed in the manufacturing of clothing and shoes, sector in which Italy is a world leader. To sustain Italian products against foreign competition in the globalization era particular attention is to be focus on product quality. Here we investigate the association of SNP mutations in genes coding for collagen proteins present in animal skin with a number of phisico-chemical parameters influencing skin quality for the tanning industry.Skin and blood were sampled from 73 Italian Friesian and Italian Brown bovines and from 43 Bergamasca and Sarda ovines, classified by sex and age. Skins were characterised for a set of chemico-physical parameters (thickness, density, humidity, protein content, ashes, lipid content, hydrossi-proline and DNA content).Regions of the collagen type I, III and IV were screened for SNP discovery in the two species by sequencing a set of reference animals. In bovine 15 polymorphisms were identified: (2 in collagen type I, 9 in collagen type III alp..

TRACEABILITY OF FOUR EUROPEAN PROTECTED GEOGRAPHIC INDICATION (PGI) BEEF PRODUCTS USING SINGLE NUCLEOTIDE POLYMORPHISMS (SNP) AND BAYESIAN STATISTICS

The use of SNPs in combination with Bayesian statistics for the geographic traceability of cattle were evaluated using a dataset comprising 24 breeds from Italy,France,Spain, Denmark, the Netherlands,Switzerland and UK genotyped with 90 polymorphic markers. The percentage of correct assignment of the individuals to their Country of origin was 90%, with an average assignment probability of 93% and an average specificity of 92%. The higher value was observed for UK breeds (97% of correct assignment) while Swiss animals were the most difficult to allocate (77% of correct assignment). Tracing of Protected Geographic Indication (PGI) products, the approach correctly assigned 100% of Guaranteed Pure Highland Beef; 97% of “Vitellone dell’Appennino Centrale” breeds; 84% of Ternera de Navarra, and 80% of Boeuf de Chalosse. Methods to verify Products of Designated Origin (PDO) and Protected Geographic Indication (PGI) products will help to protect regional foods and promote the economic growth of marginal rural areas by encouraging the product on of high quality niche market foods

Standard and Light-Cycler PCR methods for animal DNA species detection in animal feedstuffs

In this work four species-specific primers and probes were designed and evaluated for the detection and quantification of bovine, ovine, swine and chicken mitochondrial DNA in feeds. PCR primers were optimized using conventional and Real Time PCR, to detect short species-specific sequences amplifiable from heat treated material. Both methods confirmed the high specificity of the primers designed. Real time quantitative PCR assay allowed the detection of as few as 0.01 ng and 0.05 ng of ovine and bovine genomic DNA, respectively. The detection limit for swine and chicken genomic DNA was 0.5 ng. Sensitivity levels observed in DNA extracted from meat samples processed according to EU legislation were different compared to those in genomic DNAs previously described. They resulted in swine 5 fg of MBM DNA, in chicken 25 ng, in ovine and bovine 50 ng. We confirmed the efficiency and specificity of primers in RT-PCR to detect 0.5% of bovine, ovine, swine and chicken MBM in contaminated feedstuffs. (C) 2007 Elsevier Ltd. All rights reserved

Ion Pair Binding in the Solid-State with Ditopic Crown Ether Uranyl Salophen Receptors

Two ditopic uranyl salophen receptors with benzo-15-crown-5 and benzo-18-crown-6 units (<b>R</b>^<b>1</b> and <b>R</b>^<b>2</b>, respectively) have been synthesized from commercially available starting materials. Comprehensive studies on the solid-state ion pair complexation with various alkali and ammonium halides have been conducted. From the 19 obtained solid-state structures (6 structures with <b>R</b>^<b>1</b>, 13 structures with <b>R</b>^<b>2</b>), three general interaction motifs <b>I</b>–<b>III</b> have been observed. Interaction motif <b>I</b> has a separated ion pair with the cation coordinated to the crown ether unit, and the anion or oxygen containing solvent molecule coordinated to the uranyl center. The interaction motif <b>II</b> manifests a polymeric structure with a contact ion pair between the uranyl-coordinated anion and cation coordinated in the crown ether in the adjacent receptor. Interaction motif <b>III</b> consists of a more general stacked packing structure of the receptors with or without ion pairs. From the obtained solid-state structures, the complex <b>R</b>^<b>2</b><b>·NaI</b> shows an interesting formation of infinite coordination polymeric structure where the crown ether complexed sodium cations and the OUO units of the adjacent receptors form a nearly linear 1-D chains. In the course of this work also the first solid-state structure of uranyl salophen acetate complex was obtained (<b>R</b>^<b>2</b><b>·KAcO</b>)

Molecular detection of cell line cross-contaminations using amplified fragment length polymorphism DNA fingerprinting technology

We have tested amplified fragment length polymorphism (AFLP) technology, in comparison with isoenzyme analysis, for the simultaneous detection of inter- and intraspecific cell line cross-contaminations (CCCs) in the cell line collection held at the Istituto Zooprofilattico della Lombardia e dell'Emilia Romagna. Isoenzyme analysis identified four cases of interspecific CCCs. In a single experiment, AFLP was able to identify the species of origin of all cell lines for which a reference genomic deoxyribonucleic acid was available and to detect five interspecific contaminations. Four CCCs confirmed data on isoenzymes, whereas the fifth CCC was detected in a species for which isoenzyme analysis was noninformative. In addition, AFLP was able to identify the putative source of the contaminations detected. The utility of the technology in the detection of intraspecific cell line contaminations depends on the number of cell lines that have to be distinguished in a specific species and on the availability of highly informative fingerprinting systems. In mice, a single AFLP primer pair produced 16 polymorphisms and distinguished all the 15 strains of mouse cell lines analyzed. In humans, 18 AFLPs identified 83 different profiles in the 159 cell lines analyzed. Amplified fragment length polymorphism can conveniently be applied for cell line fingerprinting in species for which hypervariable markers are not available. In species for which a highly informative multiplex of microsatellite markers is available, AFLP can still provide a useful and cheap tool for simultaneously testing inter- and intraspecific contaminations