Antioxidant capacity of oat (Avena sativa L.) extracts. 1. Inhibition of low-density lipoprotein oxidation and oxygen radical absorbance capacity

Milled oat groat pearlings, trichomes, flour, and bran were extracted with methanol and the fractions tested in vitro for antioxidant capacity against low-density lipoprotein (LDL) oxidation and R-phycoerythrin protein oxidation in the oxygen radical absorbance capacity (ORAC) assay. The oxidative reactions were generated by 2,2'-azobis(2-amidinopropane) HCl (AAPH) or Cu(2+) in the LDL assay and by AAPH or Cu(2+) + H(2)O(2) in the ORAC assay and calibrated against a Trolox standard to calculate Trolox equivalents (1 Trolox equivalent = 1 TE = activity of 1 micromol of Trolox). The antioxidant capacity of the oat fractions was generally consistent with a potency rank of pearlings (2.89-8.58 TE/g) > flour (1.00-3.54 TE/g) > trichome (1.74 TE/g) = bran (1.02-1.62 TE/g) in both LDL and ORAC assays regardless of the free radical generator employed. A portion of the oat antioxidant constituents may be heat labile as the greatest activity was found among non-steam-treated pearlings. The contribution of oat tocols from the fractions accounted for <5% of the measured antioxidant capacity. AAPH-initiated oxidation of LDL was inhibited by the oat fractions in a dose-dependent manner, although complete suppression was not achieved with the highest doses tested. In contrast, Cu(2+)-initiated oxidation of LDL stimulated peroxide formation with low oat concentrations but completely inhibited oxidation with higher doses. Thus, oats possess antioxidant capacity most of which is likely derived from polar phenolic compounds in the aleurone.     

Simplified Dilute Acetic Acid Based Extraction Procedure for Fractionation and Analysis of Wheat Flour Protein by Size Exclusion HPLC and Flow Field‐Flow Fractionation

A simple, highly efficient and reproducible two‐step extraction procedure using dilute acetic acid without (AN) and then with sonication (AS) has been developed for the fractionation of wheat flour protein. Approximately 97% of total protein was extracted from a Canadian hard red spring wheat flour; an additional 1.2% protein could be recovered by further extraction with 1% DDT and 50% 1‐propanol (AR). Size‐exclusion HPLC (SE‐HPLC) and flow field‐flow fractionation (flow FFF) showed that the AN extract, which accounted for most of the total extractable protein (AN + AS + AR), consisted primarily of monomeric protein. The AS extract was composed primarily of polymeric proteins. Flow FFF showed that AN polymeric protein, including that eluting at the SE‐HPLC void volume, showed smaller Stokes diameters than AS polymeric protein. Flow FFF profiles of AS SE‐HPLC subfractions showed that the void volume subfraction contained monomeric and small polymeric protein in addition to large polymeric protein, indicating formation of larger complexes through interaction between some or all of the components. AN and AS extracts, as well as SE‐HPLC and flow FFF fractions thereof, showed a fairly wide range of values among 12 Canadian hard red and white spring wheat cultivars. The proportion of total protein in the AS extract and in the larger sized polymeric protein fractions from SE‐HPLC and flow FFF were highly positively correlated to farinograph mixing time.     

Fusarium Head Blight: Effect of Fungal Proteases on Wheat Storage Proteins

The effect of proteolytic enzymes, associated with Fusarium head blight, on wheat storage proteins and dough functionality was studied. Fusarium damaged kernels (FDK) and sound kernels were hand-picked from F. graminearum Schwabe and F. avenaceum (Fr.) Sacc. infected samples of bread and durum wheat. Scanning electron microscopy revealed significant degradation of endosperm protein in FDK. Storage proteins from FDK and sound kernels were analyzed by SDS-PAGE, RP-HPLC, and SE-HPLC. Total storage protein was lower in FDK but no significant qualitative differences in protein were detected by either RP-HPLC or SDS-PAGE. SE-HPLC was used to follow the hydrolysis of wheat storage protein by proteolytic enzymes found in FDK and a pure culture of F. graminearum. Selective inhibition of proteolytic activity by p-chloromercuribenzoate, and not soybean trypsin inhibitor or iodoacetic acid, suggests that the F. graminearum protease is an alkaline protease. Farinograph and extensigraph curves showed that the presence of FDK decreased dough consistency and resistance to extension. The presence of FDK in flour resulted in a substantial reduction in loaf volume. The loss of dough functionality and loaf volume potential was attributed to the presence of fungal proteases.     

Listeria monocytogenes fecal shedding in dairy cattle shows high levels of day-to-day variation and includes outbreaks and sporadic cases of shedding of specific L. monocytogenes subtypes

Fecal shedding of Listeria monocytogenes poses a risk for contamination of animal feed and agricultural environments and raw food at the pre-harvest stages of food production. To be able to reduce these risks it is critical to improve understanding of the epidemiology of L. monocytogenes shedding in feces. The objective of this study was to assess the daily variability of fecal shedding and its association with individual animal (lactation number and the day of current lactation) and environmental (feed) risk factors. That was achieved by application of longitudinal daily sample collection in a herd of dairy cattle and molecular characterization of isolated L. monocytogenes. Fecal samples (25) and silage samples (2) were collected daily during two 2-week periods and one 5-day period. L. monocytogenes was isolated from 255 out of 825 (31%) fecal samples on 24 out of 33 (73%) days, and from 25 out of 66 (38%) silage samples on 16 out of 33 (48%) days. Ninety-four percent of cows excreted L. monocytogenes in feces at least once during the study period. Our data analyses indicated that (i) the prevalence and incidence risk of L. monocytogenes fecal shedding in cattle vary considerably over time, from 0 to 100%, and both are associated with contamination of silage, (ii) L. monocytogenes fecal shedding in cattle could occur as part of an outbreak or as an isolated sporadic case, (iii) L. monocytogenes subtypes associated with human infections are commonly isolated from cattle feces and silage, and (iv) a single cow can harbor more than one L. monocytogenes subtype on any given day. Although limited to a single dairy cattle herd, these findings provide a significant advancement in the understanding of the epidemiology of L. monocytogenes fecal shedding in dairy cattle.     

Separating N2O production and consumption in intact agricultural soil cores at different moisture contents and depths

Agricultural soils are a major source of the potent greenhouse gas and ozone depleting substance, N₂O. To implement management practices that minimize microbial N₂O production and maximize its consumption (i.e., complete denitrification), we must understand the interplay between simultaneously occurring biological and physical processes, especially how this changes with soil depth. Meaningfully disentangling of these processes is challenging and typical N₂O flux measurement techniques provide little insight into subsurface mechanisms. In addition, denitrification studies are often conducted on sieved soil in altered O₂ environments which relate poorly to in situ field conditions. Here, we developed a novel incubation system with headspaces both above and below the soil cores and field-relevant O₂ concentrations to better represent in situ conditions. We incubated intact sandy clay loam textured agricultural topsoil (0–10 cm) and subsoil (50–60 cm) cores for 3–4 days at 50% and 70% water-filled pore space, respectively. ¹⁵N-N₂O pool dilution and an SF₆ tracer were injected below the cores to determine the relative diffusivity and the net N₂O emission and gross N₂O emission and consumption fluxes. The relationship between calculated fluxes from the below and above soil core headspaces confirmed that the system performed well. Relative diffusivity did not vary with depth, likely due to the preservation of preferential flow pathways in the intact cores. Gross N₂O emission and uptake also did not differ with depth but were higher in the drier cores, contrary to expectation. We speculate this was due to aerobic denitrification being the primary N₂O consuming process and simultaneously occurring denitrification and nitrification both producing N₂O in the drier cores. We provide further evidence of substantial N₂O consumption in drier soil but without net negative N₂O emissions. The results from this study are important for the future application of the ¹⁵N-N₂O pool dilution method and N budgeting and modelling, as required for improving management to minimize N₂O losses.     

A Bayesian Approach to Fitting Gibbs Processes with Temporal Random Effects

We consider spatial point pattern data that have been observed repeatedly over a period of time in an inhomogeneous environment. Each spatial point pattern can be regarded as a ??snapshot?? of the underlying point process at a series of times. Thus, the number of points and corresponding locations of points differ for each snapshot. Each snapshot can be analyzed independently, but in many cases there may be little information in the data relating to model parameters, particularly parameters relating to the interaction between points. Thus, we develop an integrated approach, simultaneously analyzing all snapshots within a single robust and consistent analysis. We assume that sufficient time has passed between observation dates so that the spatial point patterns can be regarded as independent replicates, given spatial covariates. We develop a joint mixed effects Gibbs point process model for the replicates of spatial point patterns by considering environmental covariates in the analysis as fixed effects, to model the heterogeneous environment, with a random effects (or hierarchical) component to account for the different observation days for the intensity function. We demonstrate how the model can be fitted within a Bayesian framework using an auxiliary variable approach to deal with the issue of the random effects component. We apply the methods to a data set of musk oxen herds and demonstrate the increased precision of the parameter estimates when considering all available data within a single integrated analysis.     

EVALUATION OP A RECIRCULATING FRESHWATER SALMON REARING FACILITY USING CLINOPTILOLITE FOR AMMONIA REMOVAL

The Seattle Aquarium has reared coho and Chinook fingerlings since 1978 in a 36,000 liter recirculating freshwater system utilizing clinoptilolite (clino) for ammonia removal. The system has effectively produced 140,000 smolts, due in large part to the efficient removal of fish-excreted ammonia by the clino filters. Quantitative studies were performed to assess clino filter loading capacity and to establish effective recharging techniques. The operational characteristics of the clino filters and suggestions for using them in freshwater recirculating culture systems are discussed.