The distribution of anthocyanins in grape skin extracts of port wine cultivars as determined by high performance liquid chromatography

The distribution of anthocyanins in methanolic skin extracts of 16 grape cultivars used for port wine production and grown at five sites in the Douro Valley in Northern Portugal has been assessed by high performance liquid chromatography. Results are confined to the seven most readily separated and identified anthocyanins and are expressed as percentages of their total (88–99% of the total integrated area). Anthocyanins based on malvidin (Mv) predominated. Of these, Mv 3-glucoside was the major pigment (33–60%), being exceeded by Mv 3-p-coumarylglucoside (2–51%) in only two cultivars; Mv 3-acetylglucoside (1–15%) was consistently the lowest. Peonidin 3-glucoside (1–27%) was prominent in four cultivars, but delphinidin 3-glucoside (1–11%), petunidin 3-glucoside (2–11%) and cyanidin 3-glucoside (trace-6%) were of low proportions throughout. The ratio Mv 3-acetylglucoside/total Mv glucosides appeared characteristic of cultivar, independent of site, and a useful aid to identification of grape cultivars. The coloured pulp of some cultivars contained peonidin 3-glucoside as a major component, present in greater proportions than in the skin; the percentage of Mv 3-p-coumarylglucoside was lower in pulp than in skin. The identities of six of the seven anthocyanins were confirmed by fast atom bombardment mass spectrometry and the structures of the Mv derivatives were determined by nuclear magnetic resonance. The formation of formyl and acetyl anthocyanin artefacts is described.     

Prediction of viscous flow around a fully submerged appended body

This paper describes the first stages in the development and application of a finite-difference calculation procedure employing a two-equation turbulence model (k, ε), to the prediction of the flow around an appended, submerged body.Initial studies were conducted to predict the steady flow field over and behind an appendage protruding through a plane boundary layer. The procedures developed to calculate both the two- and three-dimensional wake fields are presented. For the two-dimensional case the method employed is partially-parabolic and thus takes account of lateral pressure variation but cannot predict flow separation and recirculation. The three-dimensional flow field is also non-parabolic; however, by using the two-dimensional partially-parabolic solution it is possible to decouple the pressure in the momentum equations and thus render the solution parabolic. This considerably simplifies the computation procedure compared with that of a three-dimensional partially-parabolic solution. Comparison between calculated velocities and measured data demonstrate that the technique has the potential to warrant extension to consideration of the flow around an appended body.The application of the partially-parabolic calculation procedure to an unappended axisymmetric body is described and comparison again made between predicted and measured parameters. The calculated axial variation of skin friction and boundary layer thickness are shown to be in close agreement with experimental values but the static pressure gradient and total velocity are both over predicted in the tail region.The implications of the above comparison on the application of the prediction method to submerged bodies is discussed and proposals made for further work.     

Effect of drought and irrigation on the fate of nitrogen applied to cut permanent grass swards in lysimeters: Nitrogen balance sheet and the effect of sward destruction and ploughing on nitrogen mineralisation

Fuor years after ¹⁵N labelled fertiliser nitrogen (as Ca(NO₃)₂ and equivalent to 400 kg N ha^?1) was applied to permanent grass swards growing in lysimeter monoliths, approximately one-quarter remained immobilised in soil organic matter. In the intervening years similar but non-labelled applications were made. Although differing rainfall regimes applied during the experiment had significantly affected nitrogen uptake by plants and nitrate loss in drainage, they caused no significant effect on the tracer nitrogen remaining in the soil, the ranges were 85–97 kg N ha^?1 and 79–94 kg N ha^?1 respectively for the 135 cm deep clay and silt loam soil monoliths. Labelled nitrogen unaccounted for in crop, drainage or soil was presumed to have been denitrified. These losses averaged 62 and 49 kg N ha^?1 on the clay and silt loam soils respectively; again the differing rainfall regimes caused no significant differences. The ratio between estimates of labelled nitrogen denitrified and of annual nitroux oxide loss was approximately 9:1 for both soils. The conversion from permanent grass to winter wheat in autumn 1981, involving killing and then burying the sward, resulted in no pronounced increase in net mineralisation of labelled nitrogen. However, the balance between crop uptake and the quantity leached did change. Labelled nitrogen assimilated was less for the wheat (growing without addition of fertiliser nitrogen) than for the grass in its last year, and the quantity leached was considerably greater than under grass swards that were supplied with an average rainfall distribution. Following the first wheat harvest total nitrogen leached averaged 51 and 44 kg N ha^?1 on the clay and silt loam soils respectively. Rates of nitrous oxide emissions during the autumn following sward destruction were greater than in earlier years, but this enhanced loss was of short duration. The crop clearly benefited from the succession of nitrogen applications made to grass, as grain yield and total nitrogen uptake exceeded 7 t ha^?1 and 120 kg N ha^?1 respectively on both soils. These quantities exceed the national averages for winter wheat and are also considerably greater than for crops from lysimeters which received no nitrogen fertiliser throughout the experiment.     

Body composition and feed intake of young pigs postweaning

Thirty-five female Large White pigs were weaned at 14 days of age, penned individually, given a single diet and slaughtered serially at 25, 40, 55 and 70 days of age. A control group was fed to appetite throughout while three groups were subjected to feed restriction at various ages postweaning. For the periods 25 to 40, 40 to 55, 55 to 70 days of age pigs fed to appetite gained 321 (±19.9), 532 (±35.4) and 601 (±35.5) g liveweight day^?1. Pigs restricted between 25 and 55 days, followed by refeeding from 55 to 70 days, gained 192 (±14.0), 169 (±44.8) and 508 (±81.0) g liveweight day^?1; pigs restricted between 40 and 55 days, followed by refeeding from 55 to 70 days, gained 425 (±42.8), 409 (±49.6) and 480 (±70.1) g liveweight day^?1. A group given 200 g feed day^?1 gained 42 (±4.2) g liveweight daily between 25 and 70 days. Refed groups did not differ significantly from controls in their feed intakes for the period 55 to 70 days, either in absolute intakes or in feed intake kg^?1 mean liveweight. Refed pigs grew no faster, and made no greater protein gains than controls of equivalent weight; however, lipid gains were elevated.     

Individual cell lag time distributions of Cronobacter (Enterobacter sakazakii) and impact of pooling samples on its detection in powdered infant formula

Cells of six strains of Cronobacter were subjected to dry stress and stored for 2.5 months at ambient temperature. The individual cell lag time distributions of recovered cells were characterized at 25 °C and 37 °C in non-selective broth. The individual cell lag times were deduced from the times taken by cultures from individual cells to reach an optical density threshold. In parallel, growth curves for each strain at high contamination levels were determined in the same growth conditions. In general, the extreme value type II distribution with a shape parameter fixed to 5 (EVIIb) was the most effective at describing the 12 observed distributions of individual cell lag times. Recently, a model for characterizing individual cell lag time distribution from population growth parameters was developed for other food-borne pathogenic bacteria such as Listeria monocytogenes. We confirmed this model’s applicability to Cronobacter by comparing the mean and the standard deviation of individual cell lag times to populational lag times observed with high initial concentration experiments. We also validated the model in realistic conditions by studying growth in powdered infant formula decimally diluted in Buffered Peptone Water, which represents the first enrichment step of the standard detection method for Cronobacter. Individual lag times and the pooling of samples significantly affect detection performances.     

The Contribution of 4‐Hydroxyfuranone Derivatives to the Aroma of Commercial Beers and Malts

Analysis of hot water extracts from a range of commercial malts showed that 2,5‐dimethyl‐4‐hydroxy‐3(2H)furanone (DMHF) varied from undetectable (lager malt) to 4.2 mg/litre (crystal malt), a concentration twenty‐six times the flavour threshold in water. 5‐Methyl‐4‐hydroxy‐3 (2H)furanone (MHF) was detected in all samples except one but was always well under its flavour threshold value. 2(or 5)‐Ethyl‐5(or 2)‐methyl‐4‐hydroxy‐3(2H)furanone (EMHF) was not detected in any of the samples. Fermentation of lager, ale and crystal malt extracts with an ale strain of yeast led to the appearance of EMHF in all cases as well as additional DMHF. The greatest increases in both compounds was with the ale malt. Both increases and decreases occurred in MHF concentration as a result of fermentation but final levels were always well below the flavour threshold value. Analysis of ten commercial beers found DMHF in all the samples and in five cases levels exceeded twice the flavour threshold value in beer with flavour units from 2.4 to 9.1. A flavour panel noted that in four of these cases the beer had a distinctly sweet/caramel aroma which is typical of DMHF. EMHF was undetectable in six samples, detectable, but unquantifiable, in three cases but at about 80% of the flavour threshold value in the remaining sample. MHF was found in all samples but at insignificant levels. The results show that DMHF is an important flavour compound in British ales and EMHF may make a contribution in a limited number of situations. The contribution of malt type, brewery processing and yeast strain in determining the concentration of the two 4‐hydroxyfuranones in beer remains uncertain.     

Evolution of Listeria populations in food samples undergoing enrichment culturing

The isolation of Listeria monocytogenes from food is carried out using a double enrichment. It is believed that the double enrichment can allow the overgrowth of Listeria innocua in samples where both species are present. In this study, we have evaluated the impact of overgrowth between Listeria species and strains during each step of the enrichment process. The effect of factors minimizing interactions between strains or phage inhibitory effects has also been estimated. In an artificially contaminated food undergoing enrichment, overgrowth could result from competitive interactions between Listeria spp. resulting from the production of bacteriocins and bacteriophage at high initial contamination levels (>10(4) cfu/g), but not at lower levels (50-100 cfu/g) as generally found in contaminated foods. At high levels of inoculation, the competitive effect could be reduced by solidification of the selective broths, to limit the diffusion of the inhibitors. Overgrowth resulting from differences in growth rate occurred independent of the initial contamination level. However, in naturally contaminated foods undergoing enrichment, there were no absolute correlations between growth rates or inhibitory profiles in terms of strain evolution during enrichment. In fact, Listeria strains which were predominant in the original sample in most cases remained the dominant strains at the end of the enrichment, although the relative proportion of any given strain could change significantly over the enrichment process. Additional factors which have yet to be identified impact on the evolution of Listeria in the two-step enrichment process. Analysis of strain evolution in eight naturally contaminated foods has indicated that the second enrichment step in Fraser broth can be reduced from 48 to 24 h without impacting on the recovery of L. monocytogenes. Our limited survey of naturally contaminated foods also demonstrated that maximum recovery of L. monocytogenes and other Listeria strains was found following 24 h incubation in 1/2 Fraser Broth. This finding suggests that it may be possible to shorten the current two-step isolation method further without reducing method sensitivity.     

Effect of waxy flour blends on dough rheology and bread quality

Although much research has been conducted on wheat flour dough rheology, the principal focus has been the role of the protein fraction. Starch is the main component of flour and plays a key role in dough dynamic properties, particularly during heating. This study assesses the effect of two different waxy flours, a durum and a bread wheat, and their blends with commercial bakers' flour on dough rheology during heating with a concurrent investigation into baking performance. Both waxy flour blends produced similar effects on dough rheological behaviour despite differences in protein content, acting to delay gelatinisation and reduce storage modulus. The main effects in bread were to increase loaf expansion during baking and reduce loaf firmness. It is postulated these effects are largely water mediated, with the higher swelling ability of the waxy starch granules reducing overall water availability and driving complete gelatinisation to higher temperatures.     

Novel conjugative plasmids from the natural isolate Lactococcus lactis subspecies cremoris DPC3758: a repository of genes for the potential improvement of dairy starters

A collection of 17 natural lactococcal isolates from raw milk cheeses were studied in terms of their plasmid distribution, content, and diversity. All strains in the collection harbored an abundance of plasmids, including Lactococcus lactis ssp. cremoris DPC3758, whose 8-plasmid complement was selected for sequencing. The complete sequences of pAF22 (22,388 kb), pAF14 (14,419 kb), pAF12 (12,067 kb), pAF07 (7,435 kb), and pAF04 (3,801 kb) were obtained, whereas gene functions of technological interest were mapped to pAF65 (65 kb) and pAF45 (45 kb) by PCR. The plasmids of L. lactis DPC3758 were found to encode many genes with the potential to improve the technological properties of dairy starters. These included 3 anti-phage restriction/modification (R/M) systems (1 of type I and 2 of type II) and genes for immunity/resistance to nisin, lacticin 481, cadmium, and copper. Regions encoding conjugative/mobilization functions were present in 6 of the 8 plasmids, including those containing the R/M systems, thus enabling the food-grade transfer of these mechanisms to industrial strains. Using cadmium selection, the sequential stacking of the R/M plasmids into a plasmid-free host provided the recipient with increased protection against 936- and c2-type phages. The association of food-grade selectable markers and mobilization functions on L. lactis DPC3758 plasmids will facilitate their exploitation to obtain industrial strains with enhanced phage protection and robustness. These natural plasmids also provide another example of the major role of plasmids in contributing to host fitness and preservation within its ecological niche.