Occurrence of Major Whey Proteins in the pH 4.6 Insoluble Protein Fraction from UHT-Treated Milk

A clear picture of the protein rearrangement in milk following UHT-treatment was drawn by a comparative analysis of the pH 4.6 soluble protein fraction (SPF) and the pH 4.6 insoluble protein fraction (IPF) recovered from raw and UHT-treated milk samples. The two protein fractions were analyzed by mono- or bidimensional gel electrophoresis under reducing and nonreducing conditions, and protein bands were identified by specific immunostaining. Results showed that bovine serum albumin, β-lactoglobulin, and, to a lesser extent, α-lactalbumin coprecipitated with caseins in UHT-treated milk samples at pH 4.6. These proteins were almost exclusively involved in high molecular weight aggregates held together by disulfide bonds. Partition of α-lactalbumin and bovine serum albumin in the protein fractions obtained upon acidification of milk at pH 4.6 was evaluated by competitive immunoassays. The ELISA-based results suggested the possibility of using pH 4.6 insoluble α-lactalbumin and bovine serum albumin, in addition to pH 4.6 insoluble β-lactoglobulin, as indicators of the intensity of the heat treatment applied to milk.     

Quantitative trait loci for yellow pigment concentration and individual carotenoid compounds in durum wheat

A defining factor for the commercial value of durum wheat pasta is its amber colour, which depends on the semolina yellow pigment concentration and on the oxidative enzymatic activity. Among carotenoids controlling yellow colour, the presence of β-carotene is also important as precursors of vitamin A. The aim of the present study was to detect quantitative trait loci (QTL) for yellow pigment concentration, yellow index and individual carotenoid compounds (lutein, zeaxanthin, β-cryptoxanthin, α-carotene and β-carotene) in a durum segregant population. Total carotenoid concentration amounted to 37% of the yellow pigments, indicating unknown colour-producing compounds in the durum extracts. Lutein was the most abundant carotenoid, followed by zeaxanthin, α-carotene and β-carotene, while β-cryptoxanthin was a minor component. Phytoene synthase marker Psy-A1, 150 SSR and EST-SSR markers, and 345 DArT^® markers, were used to construct the linkage map for subsequent QTL analysis. Clusters of QTL for total and/or one or more carotenoid compounds were detected on the same chromosome regions (2A, 3B, 5A and 7A) where QTL for yellow pigment concentration and yellow index were identified. The molecular markers associated to major QTL would be useful for marker-assisted selection programs to facilitate high carotenoid concentration with high nutritional carotenoid compounds in wheat grain.     

Genetic diversity of a <Emphasis Type="Italic">Coffea</Emphasis> Germplasm Collection assessed by RAPD markers

Genetic diversity and relationships within and among nine species of Coffea, one species of Psilanthus and the Piatã hybrid from the Coffee Germplasm Collection of Instituto Agronômico de Campinas (IAC), Brazil were assessed using RAPD markers. Genetic diversity and relationships were evaluated by proportion of polymorphic loci (P), Shannon’s genetic index (H′ and G′_ST) and clustering analysis. The overall RAPD variation among all accessions was mostly partitioned between rather than within species. However, C. canephora and C. liberica showed a high genetic diversity within the species (\({\underline{\hbox{H}'}} \) _sp = 0.414 and \({\underline{\hbox{H}'}} \) _sp = 0.380, respectively) and this was highly structured (high \({\underline{\hbox{G}'}} \) _ST). Genetic diversity from C. congensis and C. arabica was also structured, but with lower levels of genetic diversity (\({\underline{\hbox{H}'}} \) _sp = 0.218 and \({\underline{\hbox{H}'}} \) _sp = 0.126, respectively). The results were consistent with agronomic and molecular studies and demonstrated that the IAC Coffea Collection is representative of the phylogenetic structure observed in the genera. This study devises sampling strategies for coffee germplasm collections and provides genetic diversity parameters for future comparisons among them.     

Immunochemical detection of formylated gamma(2)-casein in cheese

An immunochemical approach has been developed to detect the use of formaldehyde as a bacteriostatic agent in dairy products. A synthetic peptide, reproducing the first five amino acid residues of the gamma(2)-casein sequence, was formylated to generate the novel haptenic structure, already well-recognized in formaldehyde-treated milk and arising out of molecular rearrangement after the addition of formaldehyde to the alpha-amino group of the histidine residue at the N terminus of gamma(2)-casein. A polyclonal antibodies preparation produced against the formylated peptide adduct proved to be a highly specific analytical tool for detecting the formylated adduct of gamma(2)-casein in formaldehyde-treated milk. Polyclonal antibodies obtained against the unmodified peptide were able to detect selectively residual native gamma(2)-casein in ripened cheese.     

Giant Galápagos tortoises; molecular genetic analyses identify a trans-island hybrid in a repatriation program of an endangered taxon

Background

Giant Galápagos tortoises on the island of Española have been the focus of an intensive captive breeding-repatriation programme for over 35 years that saved the taxon from extinction. However, analysis of 118 samples from released individuals indicated that the bias sex ratio and large variance in reproductive success among the 15 breeders has severely reduced the effective population size (N _e ).

Results

We report here that an analysis of an additional 473 captive-bred tortoises released back to the island reveals an individual (E1465) that exhibits nuclear microsatellite alleles not found in any of the 15 breeders. Statistical analyses incorporating genotypes of 304 field-sampled individuals from all populations on the major islands indicate that E1465 is most probably a hybrid between an Española female tortoise and a male from the island of Pinzón, likely present on Española due to human transport.

Conclusion

Removal of E1465 as well as its father and possible (half-)siblings is warranted to prevent further contamination within this taxon of particular conservation significance. Despite this detected single contamination, it is highly noteworthy to emphasize the success of this repatriation program conducted over nearly 40 years and involving release of over 2000 captive-bred tortoises that now reproduce in situ. The incorporation of molecular genetic analysis of the program is providing guidance that will aid in monitoring the genetic integrity of this ambitious effort to restore a unique linage of a spectacular animal.