Real-time TaqMan polymerase chain reaction detection and quantification of cow DNA in pure water buffalo mozzarella cheese: method validation and its application on commercial samples

Mozzarella cheese obtained from buffalo (Bubalus bubalis) milk is a typical Italian product certificated by means of the European Protected Designation of Origin (PDO). Mozzarella cheese can also be obtained from bovine milk or bovine/buffalo milk mixtures, but in this case, it cannot be sold as PDO product, and its label must report the actual ingredients. However, bovine milk in PDO products was frequently detected in the past, suggesting fraudulent addition or accidental contamination. Several methods based on end-point polymerase chain reaction (PCR) have been profitably applied in a large number of tests to detect the presence of undeclared ingredients, also in dairy products. In the present study we report a real-time PCR method able to quantify bovine milk addition to pure buffalo cheese products. We validated a normalized procedure based on two targets: bovine mitochondrial cytochrome b (cyt b) to detect and quantify the bovine DNA and nuclear growth hormone (GH) gene used as a universal reference marker. With the use of this real-time PCR assay, 64 commercial mozzarella di bufala cheese samples purchased at local supermarkets, dairy shops, or directly from cheese manufacturers were analyzed. The results obtained demonstrate that most of the commercial samples were contaminated with bovine milk. Therefore, this assay could be conveniently employed to carry out routine and accurate controls aimed not only to discourage any fraudulent behavior but also to reduce risks for consumer health.     

Fungizidleistung von Wirkstoffgruppen mit unterschiedlichen Mode of action gegen Braunrost (Puccinia recondita f.sp.secalis Rob.ex Desm.) in Winterroggen

The economically most damaging disease to winter rye is brown rust (Puccinia recondita f.sp.secalis Rob.ex Desm.). There are fungicides of different drug groups in order to fight brown rust in agriculture. Usually mixtures of different active ingredients with different modes of action can be used for the treatment of fungal pathogens. In experiments single agents like pyrazole-carboxamides, azoles and strobilurines were compared to drug combination products like azole strobilurin, azole-carboxamide, or azole carboxamide-strobilurin mixture and their fungicide performance was determined. The active ingredients Epoxiconazole, Pyraclostrobin, and Fluxapyroxad were tested. In eight trials from 2012 to 2013 Fluxapyroxad reached the highest levels of efficiency for P. recondita. Pyraclostrobin or the combination of Pyraclostrobin and Epoxiconazole showed higher efficiencies than Epoxiconazole or the combination of Epoxiconazole and Metconazole in all trials. The thousand seed weight and income from the harvest reflect the efficiency against P. recondita. Both azole strobilurin mixtures as well as the azole carboxamide mixtures are suitable for the control of leaf rust in agriculture. In order to minimize the risk of resistance to strobilurin and azole carboxamide mixtures should be used alternately.     

Census Report of the <Emphasis Type="BoldItalic">Potato Virus Y</Emphasis> (PVY) Population in Swiss Seed Potato Production in 2003 and 2008

The status of the Potato virus Y (PVY) in Swiss seed potato production was investigated in the years 2003 and 2008 by analysing 385 leaf samples of field-grown, suspicious potato plants collected in four representative seed control fields. Serological investigations by ELISA showed that in c. 84% of the PVY-positive samples in both years, viruses belonging to the PVY^N group were found. All 124 serologically positive PVY samples collected in 2003 and a selection of 81 isolates of 2008 were further typified by molecular tests and by biological assays on tobacco and potato plants. These tests largely confirmed the predominance of the PVY^N group and, within this group, the prevalence of recombinant PVY^NTN, with 81.4% and 70.4% in 2003 and 2008, respectively. The percentage of PVY^N-Wilga (PVY^N-Wi) increased from c. 6% to 17% between the two years. PVY^O was detected only in 10.5% and 4.9% of all molecularly analysed samples in 2003 and 2008, respectively. The persistent predominance of recombinant PVY^NTN in Swiss seed potatoes indicates that this strain group is now widespread, representing a considerable threat to Swiss seed potato production.     

Genome-wide analysis of plastome sequence variation and development of plastidial CAPS markers in common potato and related <Emphasis Type="Italic">Solanum</Emphasis> species

The plastome sequence of the European cultivated potato, Solanum tuberosum subsp. tuberosum (tbr, GenBank accession no. DQ386163), was compared with that of S. bulbocastanum, a wild potato relative (blb, GenBank accession no. DQ347958), in order to characterize the degree and type of variability in different genomic regions, and develop molecular markers relevant to genetics, breeding and biotechnology of potato. One hundred forty-two and 251 PICs (Potentially Informative Characters) were found in coding and non-coding sequences (NCSs), respectively. Further, while variation in coding regions was almost exclusively due to nucleotide substitutions, 25% of PICs in NCSs of tbr and blb were due to indels, most of them mononucleotide or longer tandem repeats (micro and minisatellites). Four intergenic regions were selected for further analyses in other 16 tuber-bearing Solanum species. The rps16-trnQ ^UUG gene spacer was found to be the most variable, forty-six PICs in this region distinguishing 18 haplotypes. Analysis of haplotype relationships, based on variability in the four intergenic regions, confirmed that the most primitive species from Central America were the most distant to S. tuberosum. Finally, polymorphic sites in the same regions were used to develop a set of CAPS (Cleaved Amplified Polymorphic Sequences) markers for species/cytoplasm identification in Solanum spp.     

The low-molecular-weight glutenin subunits of wheat gluten

Low-molecular-weight glutenin subunits (LMW-GS) are polymeric protein components of wheat endosperm and like all seed storage proteins, are digested to provide nutrients for the embryo during seed germination and seedling growth. Due to their structural characteristics, they exhibit features important for the technological properties of wheat flour. Their ability to form inter-molecular disulphide bonds with each other and/or with high-molecular-weight glutenin subunits (HMW-GS), is important for the formation of the glutenin polymers, which are among the biggest macromolecules present in nature, and determine the processing properties of wheat dough. Explanation of the structural basis for these correlations continues to intrigue researchers and, while earlier emphasis had been on HMW-GS, considerable attention is now being focused on the LMW-GS.LMW-GS are a highly polymorphic protein complex, including proteins with gliadin-type sequences. Difficulty in separating single components, arising from the complexity of the group, has limited the characterisation of the individual proteins and the establishment of clear-cut relationships with quality parameters.Here we review results concerning different aspects of LMW-GS, including their structural characteristics, genetic control, and relationships with quality parameters. In addition, we emphasise the distinction between the components with sequences unique to the LMW-GS fraction and those behaving like glutenin subunits (incorporated into polymers), but with sequences corresponding to gliadins.     

Biochemical Characterization of Cassiopea andromeda (Forsskål, 1775), Another Red Sea Jellyfish in the Western Mediterranean Sea

Increasing frequency of native jellyfish proliferations and massive appearance of non-indigenous jellyfish species recently concur to impact Mediterranean coastal ecosystems and human activities at sea. Nonetheless, jellyfish biomass may represent an exploitable novel resource to coastal communities, with reference to its potential use in the pharmaceutical, nutritional, and nutraceutical Blue Growth sectors. The zooxanthellate jellyfish Cassiopea andromeda, Forsskål, 1775 (Cnidaria, Rhizostomeae) entered the Levant Sea through the Suez Canal and spread towards the Western Mediterranean to reach Malta, Tunisia, and recently also the Italian coasts. Here we report on the biochemical characterization and antioxidant activity of C. andromeda specimens with a discussion on their relative biological activities. The biochemical characterization of the aqueous (PBS) and hydroalcoholic (80% ethanol) soluble components of C. andromeda were performed for whole jellyfish, as well as separately for umbrella and oral arms. The insoluble components were hydrolyzed by sequential enzymatic digestion with pepsin and collagenase. The composition and antioxidant activity of the insoluble and enzymatically digestible fractions were not affected by the pre-extraction types, resulting into collagen- and non-collagen-derived peptides with antioxidant activity. Both soluble compounds and hydrolyzed fractions were characterized for the content of proteins, phenolic compounds, and lipids. The presence of compounds coming from the endosymbiont zooxanthellae was also detected. The notable yield and the considerable antioxidant activity detected make this species worthy of further study for its potential biotechnological sustainable exploitation.     

Characterization of Phaseolus vulgaris L. Landraces Cultivated in Central Italy

Eight Phaseolus vulgaris L. landraces cultivated on farm in marginal areas of Central Italy (Lazio region) were investigated in order to evaluate chemical composition of storage proteins and secondary metabolites fractions. The total protein content showed some differences among landraces; the maximum value was next to 30 g for 100 g of dry weight. The seed storage proteins were screened by polyacrylamide gel electrophoresis (SDS/PAGE): seven landraces exhibited phaseolin patterns type S, one landrace showed a phaseolin pattern type T. A morphological analysis of cotyledon parenchyma performed by scanning electron microscopy (SEM) revealed differences in size of starch granules. Moreover the polyphenolic composition was investigated using HPLC-APCI; from the methanol extracts a flavonoid, kaempferol, and a coumarin, 5,7-dimethoxycoumarin, were identified. To our knowledge, this is the first time that 5,7-dimethoxycoumarin has been reported in P. vulgaris seeds.     

New molecular markers linked to qualitative and quantitative powdery mildew and scald resistance genes in barley for dry areas

A partial genetic linkage map was constructed on 71 doubled-haploid lines derived from a cross between the barley lines Tadmor and WI2291 with 181 molecular markers. The segregating population was used to detect markers linked to the gene Mlg conferring resistance to powdery mildew (Erysiphe graminis f. sp. hordei) and to genes for quantitative resistance to scald (Rhynchosporium secalis). The gene Mlg on chromosome 4H was flanked by two AFLP markers at a distance of 2.0 and 2.4 cM, respectively. QTLs for resistance to scald were detected on chromosomes 2H and 3H. This association of molecular markers with qualitative and quantitative disease resistance loci represents a valuable starting-point for marker-assisted selection. This revised version was published online in July 2006 with corrections to the Cover Date.