Salmonella carriage in an Irish pig herd: correlation between serological and bacteriological detection methods

Salmonella carriage in pigs represents a serious health problem that undoubtedly contributes to the spread of human disease. Thus, the efficient and reliable testing of farm animals for bacteria such as Salmonella is an important aspect of any efficient control strategy. Serological analysis of 15 meat juice samples detected antibodies against Salmonella in some. but not all, of the animals identified bacteriologically as harboring the pathogen, indicating a lack of correlation between the bacteriological and serological methods used for Salmonella detection. The results suggest that testing by enzyme-linked immunosorbent assay is appropriate at the herd level, with culture methods preferable for individual animal analysis. A novel culture protocol detected Salmonella in the cecal contents of 15 pigs, whereas a method based on the European Standard identified only 9 pigs as being Salmonella-positive. During the study, an unusual finding was the relatively high incidence of Salmonella London carriage in the pigs tested.     

Tandem mass spectrometry of sulfated heparin-like glycosaminoglycan oligosaccharides

The structural characterization of heparin-like glycosaminoglycans (HLGAGs) is a major challenge in glycobiology. These linear, sulfated oligosaccharides are expressed on animal cell surfaces, in extracellular matrixes, basement membranes, and mast cell granules and bind with varying degrees of specificity to families of proteases, growth factors, chemokines, and blood coagulation proteins. Cell surface HLGAGs bind growth factors and growth factor receptors and serve as coreceptors in these interactions. Understanding of the mechanism and regulation of growth factor-receptor binding requires efficient determination of cell surface HLGAG structures and the variations in their expression in response to the cellular environment. The solution to this problem entails rapid, sensitive structural analysis of these molecules. To date, HLGAG sequencing requires multistep processes that combine chemical and enzymatic degradation with gel-based or mass spectrometry-based detection systems. Although tandem mass spectrometry has revolutionized proteomics, the fragility of sulfate groups has limited its usefulness in the analysis of HLGAGs. This work demonstrates that tandem mass spectrometry can be effectively used to determine HLGAG structures while minimizing losses of SO3. First, collision-induced dissociation (CID) is shown to produce abundant backbone cleavage ions for HLGAG oligosaccharides, provided that most sulfate groups are deprotonated. Fragmentation of different precursor ion charge states produces complementary data on the structure of the HLGAG. Second, calcium ion complexation of HLGAGs stabilizes the sulfate groups, increases the relative abundances of backbone cleavage ions, and decreases the abundances of ions produced from SO3 losses.     

Further characterization of rat dihydroceramide desaturase: Tissue distribution, subcellular localization, and substrate specificity

The introduction of the double bond in the sphingoid backbone of sphingolipids occurs at the level of dihydroceramide via an NADPH-dependent desaturase, as discovered in permeabilized rat hepatocytes. In the rat, the enzyme activity, which has now been further characterized, appeared to be mostly enriched in liver and Harderian gland. By means of subcellular fractionation of rat liver homogenates and density gradient separation of microsomal fractions, the desaturase was localized to the endoplasmic reticulum. Various detergents were inhibitory to the enzyme, and maximal activities were obtained in the presence of NADPH and when the substrate was complexed to albumin. In the presence of albumin, the chain length of the fatty acid of the truncated dihydroceramides hardly affected the activity. Finally, in view of a likely evolutionary relationship between desaturases and hydroxylases, the formation of hydroxylated intermediates was analyzed. No evidence for their presence was found under our assay conditions. In part presented at the Conférence Jacques Monod “Cell lipids: Topology, transport and signalling functions”, Aussois, May 1997 (France); Van Veldhoven, P.P. “Ceramide biosynthesis: Characterisation of the conversion of dihydroceramide to ceramide”. Contributed equally to this study.     

Comprehensive survey of pasteurized fluid milk produced in the United States reveals a low prevalence of Listeria monocytogenes

A comprehensive survey was undertaken to generate contemporary data on the prevalence of Listeria monocytogenes in pasteurized fluid milk produced in the United States. Samples (5,519) near the sell-by expiration date were purchased at retail outlets over a 5-week period and analyzed for presence of L. monocytogenes. Products consisted of whole milk, nonfat milk, and chocolate milk packaged in gallon, half gallon, quart, pint, and half-pint containers. Samples were collected from both large and small retail stores in urban and suburban locations in four FoodNet cities (Baltimore, Md., Atlanta, Ga., St. Paul/ Minneapolis, Minn., and San Francisco, Calif.). Samples were prescreened for L. monocytogenes by the AOAC-approved rapid Vitek immunodiagnostic assay system, enzyme-linked fluorescent assay method. Positive prescreening samples were cultured according to the Bacteriological Analytical Manual, enumerated for L. monocytogenes with a nine-tube most-probable-number (MPN) procedure, and confirmed by biochemical characterization. The frequency of isolation of L. monocytogenes in these products was 0% (0 of 1,897) in whole milk, 0.05% (1 of 1,846) in nonfat milk, 0% (0 of 1,669) in chocolate milk, and 0% (0 of 107) in other (reduced fat and low fat) milk samples. Overall, L. monocytogenes was confirmed in only 0.018% of pasteurized milk samples (1 of 5,519). Enumeration of the single confirmed positive nonfat milk sample revealed low-level contamination (<0.3 MPN/g), even when sampled 5 days past the expiration of the sell-by date. The results confirm the low frequency of contamination of pasteurized fluid milk products by L. monocytogenes for products sold in the United States and reaffirm the reduction of contamination frequency of fluid milk by L. monocytogenes when compared with earlier estimates from the U.S. Food and Drug Administration Dairy Safety Initiatives Program.     

Influence of air drying temperature on kinetics,physicochemical properties,total phenolic content and ascorbic acid of pears

This study was conducted to evaluate quality and structural changes in parallelepipedic pieces of pears during convective drying at different air temperatures (30–70 °C).Submitted to atmospheric O₂ conditions, ascorbic acid deterioration demonstrated first-order kinetic behaviour and was found to depend on air temperature and pear moisture content. Loss of ascorbic acid content increased with increasing air temperature. Possible explanation could be the irreversible oxidative reaction occurring during drying. Phenol content degradation fitted a pseudo first-order reaction and was significantly influenced by air temperature.Variations in bulk density, shrinkage and porosity essentially depended on changes in moisture content. Porosity exhibited a nonlinear variation with respect to moisture content. Volume change showed, as expected, a linear variation with moisture content. Drying temperature significantly induced the increase of a* and b* colorimetric parameters due to non-enzymatic browning reaction, which turned the samples more reddish and yellow when the temperature rose.     

Increased Prostaglandin Response to Oxytocin in Ewes Fed a Diet High in Omega-6 Polyunsaturated Fatty Acids

Diets high in omega-6 polyunsaturated fatty acids (n-6) are associated with increased prostaglandin F_2α (PGF_2α) synthesis in cattle, however, the specific effects on the potential prostaglandin response to an oxytocin challenge in sheep have not been reported. The aim of the current study was to determine whether oxytocin-stimulated PGF_2α was significantly increased when ewes were fed a diet high in n-6 compared with a control diet low in n-6. Merino x Border Leicester ewes (n = 30) received one of two dietary treatments, either high in n-6 (70 % oat grain) or low in n-6 (control diet, 100 % cereal/legume silage). Ewes consumed the diets for 44 days prior to two consecutive oxytocin challenges. Plasma n-6 and PGF_2α metabolite (PGFM) concentrations following oxytocin challenge were greater (P < 0.05) when ewes were fed a diet high in n-6 compared with the control diet. A higher availability of n-6 may have lead to an increased in vivo synthesis of PGF_2α, however, further research is required to determine the exact mechanisms involved.