Metabolomic profiling of wild rooibos (Aspalathus linearis) ecotypes and their antioxidant-derived phytopharmaceutical potential.

INTRODUCTION: Aspalathus linearis (commonly known as rooibos) is endemic to the Cape Floristic Region of South Africa and is a popular herbal drink and skin phytotherapeutic ingredient, with health benefits derived primarily from its unique phenolic content. Several, seemingly habitat-specific ecotypes from the Cederberg (Western Cape) and Northern Cape have morphological, ecological, genetic and biochemical differences. OBJECTIVES AND METHODS: Despite the commercial popularity of the cultivated variety, the uncultivated ecotypes are largely understudied. To address gaps in knowledge about the biochemical constituency, ultra-performance liquid chromatography-mass spectrometry analysis of fifteen populations was performed, enabling high-throughput metabolomic fingerprinting of 50% (v/v) methanolic extracts. Antioxidant screening of selected populations was performed via three assays and antimicrobial activity on two microbial species was assessed. The metabolomic results were corroborated with total phenolic and flavonoid screening of the extracts. RESULTS AND DISCUSSION: Site-specific chemical lineages of rooibos ecotypes were confirmed via multivariate data analyses. Important features identified via PLS-DA disclosed higher relative abundances of certain tentative metabolites (e.g., rutin, aspalathin and apiin) present in the Dobbelaarskop, Blomfontein, Welbedacht and Eselbank sites, in comparison to other locations. Several unknown novel metabolites (e.g., m/z 155.0369, 231.0513, 443.1197, 695.2883) are responsible for metabolomic separation of the populations, four of which showed higher amounts of key metabolites and were thus selected for bioactivity analysis. The Welbedacht and Eselbank site 2 populations consistently displayed higher antioxidant activities, with 2,2-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging activities of 679.894 ± 3.427 µmol Trolox/g dry matter and 635.066 ± 5.140 µmol Trolox/g dry matter, respectively, in correlation with a high number of phenolic and flavonoid compounds. The contribution of the individual metabolites to the pharmacological effectiveness of rooibos remains unknown and as such, further structural elucidation and phytopharmacological testing is thus urgently needed.

Resprouters Versus Reseeders: Are Wild Rooibos Ecotypes Genetically Distinct?

Aspalathus linearis (Burm. F.) R. Dahlgren (Fabaceae) or rooibos, is a strict endemic species, limited to areas of the Cederberg (Western Cape) and the southern Bokkeveld plateau (Northern Cape) in the greater Cape Floristic Region (CFR) of South Africa. Wild rooibos, unlike the cultivated type, is variable in morphology, biochemistry, ecology and genetics, and these ecotypes are broadly distinguished into two main groups, namely, reseeders and resprouters, based on their fire-survival strategy. No previous assessment of genetic diversity or population structure using microsatellite markers has been conducted in A. linearis. This study aimed to test the hypothesis that wild rooibos ecotypes are distinct in genetic variability and that the ecotypes found in the Northern Cape are differentiated from those in the Cederberg that may be linked to a fire-survival strategy as well as distinct morphological and phytochemical differences. A phylogeographical and population genetic analyses of both chloroplast (trnLF intergenic region) and newly developed species-specific nuclear markers (microsatellites) was performed on six geographically representative wild rooibos populations. From the diversity indices, it was evident that the wild rooibos populations have low-to-moderate genetic diversity (He: 0.618-0.723; Ho: 0.528-0.704). The Jamaka population (Cederberg, Western Cape) had the lowest haplotype diversity (H = 0.286), and the lowest nucleotide diversity (π = 0.006) even though the data revealed large variations in haplotype diversity (h = 0.286-0.900) and nucleotide diversity (π = 0.006-0.025) between populations and amongst regions where wild rooibos populations are found. Our data suggests that populations of rooibos become less diverse from the Melkkraal population (Suid Bokkeveld, Northern Cape) down towards the Cederberg (Western Cape) populations, possibly indicative of clinal variation. The largest genetic differentiation was between Heuningvlei (Cederberg, Western Cape) and Jamaka (FST = 0.101) localities within the Cederberg mountainous region, and, Blomfontein (Northern Cape) and Jamaka (Cederberg) (FST = 0.101). There was also a significant isolation by distance (R2 = 0.296, p = 0.044). The presence of three main clusters is also clearly reflected in the discriminant analysis of principal components (DAPC) based on the microsatellite marker analyses. The correct and appropriate management of wild genetic resources of the species is urgently needed, considering that the wild Cederberg populations are genetically distinct from the wild Northern Cape plants and are delineated in accordance with ecological functional traits of reseeding or resprouting, respectively. The haplotype divergence of the ecotypes has also provided insights into the genetic history of these populations and highlighted the need for the establishment of appropriate conservation strategies for the protection of wild ecotypes.

Effect of vitamin D, zilpaterol hydrochloride supplementation, and postmortem aging on shear force measurements of three muscles in finishing beef steers.

Vitamin D (D3) supplementation may be used to increase tenderness in beef from cattle fed zilpaterol hydrochloride (ZH). The study was arranged as a 2 × 2 factorial with fixed effects of ZH (no ZH or ZH fed at 8.3 mg/kg DM for 20 d with a 3-d withdrawal) and D3 (no D3 or 500,000 IU D3·steer·d for 10 d prior to harvest). Cattle ( = 466) were harvested in 2 blocks on the basis of BW with subsequent collection of carcass data. Full loins and inside rounds ( = 144 of each subprimal) were collected for fabrication of 5 steaks from the longissimus lumborum (LL), gluteus medius (GM), and semimembranosus (SM), which were aged for 7, 14, 21, 28, or 35 d. Warner-Bratzler shear force (WBSF) was used to evaluate mechanical tenderness of LL, GM, and SM steaks at all aging periods. Slice shear force (SSF) analysis was conducted on only 14- and 21-d LL steaks. No interactions ( > 0.05) between ZH and D3 occurred throughout the entire study. Supplementing ZH resulted in increased HCW ( < 0.01), larger LM area ( < 0.01), and improved calculated yield grades ( < 0.01) with decreases in fat thickness ( = 0.02) and marbling scores ( = 0.05). Supplementation with D3 increased calculated yield grade ( < 0.01) and decreased ( = 0.01) rib eye area. Feeding ZH increased ( ≤ 0.05) WBSF of LL steaks at each postmortem age interval, whereas D3 had no effect ( > 0.05) on WBSF or SSF of LL steaks. Like for WBSF, ZH supplementation increased SSF values at 14 and 21 d postmortem ( < 0.01) compared with those for non-ZH steaks. There was an interaction between ZH and postmortem age ( < 0.01) for WBSF of LL steaks. At 7 d LL steaks from ZH steers sheared over 0.6 kg greater than non-ZH steaks; however, by 21 d this difference was reduced to an average of 0.2 kg. Differences in distribution between LL steaks below 3.0 kg from non-ZH and ZH-fed cattle were also notable ( ≤ 0.05) through 21 d of aging. At 35 d postmortem a high proportion of LL steaks (68.5%) from ZH-fed steers required less than 3.0 kg to shear. Supplementation with ZH and D3 had no impact ( > 0.05) on WBSF values of GM steaks. Feeding ZH did not alter WBSF of SM steaks, but at 28 d D3 increased ( = 0.04) WBSF values. Shear force in ZH steaks was not effectively reduced by feeding D3 for 10 d to steers prior to harvest. Aging, however, was an effective method of reducing initially greater shear force values in LL steaks and, to a lesser degree, GM steaks from ZH-fed cattle.

Changes in Sodium, Calcium, and Magnesium Ion Concentrations That Inhibit Geobacillus Biofilms Have No Effect on Anoxybacillus flavithermus Biofilms.

This study investigated the effects of varied sodium, calcium, and magnesium concentrations in specialty milk formulations on biofilm formation by Geobacillus spp. and Anoxybacillus flavithermus. The numbers of attached viable cells (log CFU per square centimeter) after 6 to 18 h of biofilm formation by three dairy-derived strains of Geobacillus and three dairy-derived strains of A. flavithermus were compared in two commercial milk formulations. Milk formulation B had relatively high sodium and low calcium and magnesium concentrations compared with those of milk formulation A, but the two formulations had comparable fat, protein, and lactose concentrations. Biofilm formation by the three Geobacillus isolates was up to 4 log CFU cm(-2) lower in milk formulation B than in milk formulation A after 6 to 18 h, and the difference was often significant (P ≤ 0.05). However, no significant differences (P ≤ 0.05) were found when biofilm formations by the three A. flavithermus isolates were compared in milk formulations A and B. Supplementation of milk formulation A with 100 mM NaCl significantly decreased (P ≤ 0.05) Geobacillus biofilm formation after 6 to 10 h. Furthermore, supplementation of milk formulation B with 2 mM CaCl2 or 2 mM MgCl2 significantly increased (P ≤ 0.05) Geobacillus biofilm formation after 10 to 18 h. It was concluded that relatively high free Na(+) and low free Ca(2+) and Mg(2+) concentrations in milk formulations are collectively required to inhibit biofilm formation by Geobacillus spp., whereas biofilm formation by A. flavithermus is not impacted by typical cation concentration differences of milk formulations.

Comparative effects of beta-adrenergic agonist supplementation on the yield and quality attributes of selected subprimals from calf-fed Holstein steers.

Mechanical portioning tests were performed on beef rib, strip loin, tenderloin, and top sirloin subprimals obtained from calf-fed Holstein steers to characterize the influence of zilpaterol hydrochloride (ZH), ractopamine hydrochloride (RH), or no ß-adrenergic agonist (ßAA; CON) on subprimal and steak yield. In addition, ßAA effects on tenderness, composition, and raw and cooked color of steaks from the aforementioned strip loin subprimals were characterized. At 14 to 15 d (ribs, tenderloins, and top sirloin) or 16 d (strip loin) postmortem, subprimals were portioned into steaks using a mechanical portioning machine. The appropriate variables were measured before and after portioning to determine ßAA influence on trimmed and untrimmed subprimal weight, subprimal length (rib only), steak weight and yield, and steak thickness (rib only). Steaks obtained from the strip loin subprimals were subjected to analysis of raw instrument color (L*, a*, b*), proximate composition, and pH. In addition, strip steaks were aged (16 or 23 d) before analysis of cooked internal color, Warner-Bratzler shear force (WBSF), and slice shear force (SSF). Briefly, ZH supplementation increased (P < 0.01) the weight of all subprimals when compared to CON. Furthermore, subprimals from CON animals consistently had fewer and lighter steaks (P ≤ 0.04) than subprimals from ZH-fed steers. Additionally, raw steaks from ZH cattle were a less vivid red (lower a* and saturation index values; P < 0.01) when compared to CON and RH steaks, which did not differ (P > 0.05). There was no interaction between ßAA treatment and postmortem aging length for WBSF or SSF (P > 0.10). However, CON steaks (3.25 kg) had lower WBSF values (P < 0.05) than ZH or RH steaks (3.68 and 3.67 kg, respectively). Regardless, aging for 23 d vs. 16 d resulted in decreased WBSF and SSF (P < 0.01) for all ßAA treatments. Although differences were numerically small, evaluations indicated the internal cooked surfaces of ZH and RH steaks were less red (P < 0.05) than CON steaks. Overall, these data reemphasize increased subprimal weights due to ßAA supplementation, particularly ZH. However, the data are not indicative of increased steak yield due to ßAA supplementation. Furthermore, the data demonstrate ßAA supplementation increases the shear force of calf-fed Holstein strip steaks regardless of postmortem aging period. However, no differences in shear force between the ßAA treatments (ZH or RH) were noted.

Comparative effects of zilpaterol hydrochloride and ractopamine hydrochloride on live performance and carcass characteristics of calf-fed Holstein steers.

Holstein steers (n = 2,275) were assigned to 1 of 3 treatments: 1) a control diet containing no ß-agonists, 2) a diet that contained zilpaterol hydrochloride (ZH; 8.3 mg/kg [100% DM basis]) for 20 d with a 3-d withdrawal period before harvest, and 3) a diet that contained ractopamine hydrochloride (RH; 30.1 mg/kg [100% DM basis]) for 28 d before harvest. No differences (P ≥ 0.18) were detected between treatments for initial BW, BW at d 28, or DMI. Final BW, BW gain for the last 28 d, total BW gain, ADG for the last 28 d, and overall ADG were greater (P < 0.05) for steers fed ZH or RH than for steers fed the control diet. Additionally, G:F for the last 28 d and G:F for the entire trial was increased (P < 0.02) for steers fed ZH (0.147, 0.147) or RH (0.153, 0.151) compared to steers fed the control diet (0.134, 0.143), respectively. Steers fed ZH or RH had HCW that were 15.5 and 8.2 kg heavier (P ≤ 0.01) and LM areas that were 7.1 and 2.3 cm(2) larger (P < 0.01) than control cattle. Steers fed ZH also had dressed carcass yields that were 1.3% to 1.5% greater and USDA calculated yield grades that were decreased 0.16 to 0.23 units compared to RH and control steers. No differences (P ≥ 0.39) were found between treatments for marbling score, fat thickness, and percentage KPH. Steers fed ZH had an increased (P ≤ 0.04) percentage of yield grade 1 and 2 carcasses (15.1, 55.0) and a reduced (P ≤ 0.02) percentage of yield grade 3 carcasses (27.1) compared with those fed RH (10.5, 49.1, 36.1) or the control diet (9.0, 47.4, 36.4), respectively. Additionally, ZH-fed steers had a decreased (P ≤ 0.04) percentage of yield grade 4 and 5 carcasses (2.8) compared with steers fed the control diet (6.9). Steers fed ZH had an increased (P ≤ 0.01) percentage of USDA Select grading carcass (31.0%) and a decreased (P ≤ 0.01) percentage of USDA Choice grading carcasses (65.0%) compared with steers fed RH (25.8%, 70.2%) and no ß-agonist (24.8%, 72.0%), respectively. Feeding either ß-agonist to calf-fed Holstein steers increased live performance through increased BW, BW gain, and ADG. Furthermore, supplementing calf-fed Holstein steers with ZH provides greater improvements in HCW, LM area, and yield grade components, with a slight decrease in quality grade when compared to calf-fed Holstein steers supplemented with RH.

Comparative effects of supplementing beef steers with zilpaterol hydrochloride, ractopamine hydrochloride, or no beta agonist on strip loin composition, raw and cooked color properties, shear force, and consumer assessment of steaks aged for fourteen or twenty-one days postmortem.

Beef steers (n = 1,914) were assigned to 1 of 3 ß-adrenergic agonist (ßAA) supplementation treatments-zilpaterol hydrochloride (ZH; 8.3 mg/kg of DM for 20 d with 3-d withdrawal), ractopamine hydrochloride (RH; 308 mg·head(-1)·d(-1) for 28 d), or no ßAA (CON)-to determine the effects on consumer eating quality. Strip loins (n = 1,101; CON = 400, RH = 355, and ZH = 346) were obtained and fabricated into 2.5-cm-thick steaks for proximate, Warner-Bratzler shear force (WBSF), slice shear force (SSF), and consumer analyses; steaks were aged until 14 or 21 d postmortem. Fat and moisture contents were not affected by ßAA supplementation (P > 0.05), but strip steaks from steers fed ZH had more protein (P < 0.01) than those from steers fed CON or RH, which were similar. An interaction between ßAA and aging was observed (P < 0.01) for WBSF but not SSF. Within steaks aged 14 d, ZH steaks required the most force to shear, RH steaks were intermediate, and CON steaks had the lowest WBSF values; however, RH steaks had a stronger response to aging than CON or ZH steaks, resulting in the lowest WBSF values at 21 d. Slice shear force values were greater (P < 0.01) in steaks from steers fed ZH than in steaks from steers fed CON or RH, which did not differ. Following shear force analyses, steaks within 2 SD of each treatment mean for WBSF were selected randomly for consumer assessment of eating quality. Consumer testing (n = 400; 200/postmortem aging period) was arranged in a 3 × 3 factorial representing 3 quality grades (Select, Low Choice, and Premium Choice) and 3 treatments (ZH, RH, and CON). In steaks aged 14 d, ßAA supplementation affected (P < 0.01) tenderness, flavor, and overall liking and tenderness acceptability, resulting in lower consumer scores for ZH than CON and RH; however, juiciness, flavor, and overall acceptability were similar (P > 0.05). In steaks aged 21 d, feeding ßAA influenced (P < 0.01) only tenderness and juiciness scores. Despite these differences, ßAA did not affect (P > 0.05) acceptability. Quality grade impacted (P < 0.01) all traits and acceptability in steaks aged 14 and 21 d. In 14-d steaks, Premium Choice typically was scored higher than Low Choice or Select; however, consumers rated 21-d Low Choice and Premium Choice similarly-both receiving greater scores than Select. Consumers detected several differences in eating quality at 14 d because of ßAA supplementation. Increasing aging from 14 to 21 d mitigated differences in shear force and tenderness scores because of feeding ZH, so that tenderness and overall acceptability were similar between ZH, RH, and CON.

Runoff quality from no-till cotton fertilized with broiler litter in subsurface bands.

Surface broadcast of broiler litter to no-till row crops exposes the litter and its nutrients to risks of loss in runoff water and volatilization and may limit the potential benefit of litter to the crops. Subsurface banding of litter could alleviate these risks. A field study was conducted in 2008 and 2009 on an upland Falkner silt loam soil to determine the effect of broiler litter placement on runoff nutrient losses from no-till cotton ( L.). Treatments included surface broadcast broiler litter applied manually, subsurface-banded litter applied by tractor-drawn equipment, and no broiler litter, all in combination with or without winter wheat ( L.) cover crop residue. Broiler litter rate was 5.6 Mg ha. The experimental design was a randomized complete block with a split-plot arrangement of treatments replicated three times. In 2008, simulated rainfall was used to generate runoff 27 d after litter application. Subsurface-banded litter reduced runoff total C, N, P, NH, NO, Cu, Zn and water-soluble P (WP) concentrations by 72, 64, 51, 49, 70, 36, 65, and 77%, respectively, compared with surface broadcast. The reductions were greater in 2009 where runoff occurred 1 d after litter application. Bacterial runoff was decreased by one log with subsurface-banded litter compared to surface broadcast. Except for C, NH, N, and WP, the presence of winter cover crop residue did not affect the load or runoff nutrient concentrations in either year. The results indicate that subsurface banding litter to no-till cotton substantially reduces nutrient and bacterial losses in runoff compared with surface broadcasting.

The effect of poultry manure application rate and AlCl(3) treatment on bacterial fecal indicators in runoff.

Increasing costs associated with inorganic fertilizer have led to widespread use of broiler litter. Proper land application, typically limiting nutrient loss, is essential to protect surface water. This study was designed to evaluate litter-borne microbial runoff (heterotrophic plate count bacteria, staphylococci, Escherichia coli, enterococci, and Clostridium perfringens) while applying typical nutrient-control methods. Field studies were conducted in which plots with high and low litter rates, inorganic fertilizer, AlCl(3)-treated litter, and controls were rained on five times using a rain generator. Overall, microbial runoff from poultry litter applied plots was consistently greater (2-5 log(10) plot(-1)) than controls. No appreciable effect on microbial runoff was noted from variable litter application rate or AlCl(3) treatments, though rain event, not time, significantly affected runoff load. C. perfringens and staphylococci runoff were consistently associated with poultry litter application, during early rain events, while other indicators were unreliable. Large microbial runoff pulses were observed, ranging from 10(2) to 10(10) CFU plot(-1); however, only a small fraction of litter-borne microbes were recoverable in runoff. This study indicated that microbial runoff from litter-applied plots can be substantial, and that methods intended to reduce nutrient losses do not necessarily reduce microbial runoff.

Effects of postmortem calcium chloride injection on meat palatability traits of strip loin steaks from cattle supplemented with or without zilpaterol hydrochloride.

An experiment was conducted to determine the effects of zilpaterol hydrochloride mM supplementation (ZH; 8.3 mg/kg on a DM basis for 20 d) and calcium chloride injection [CaCl(2), 200 at 5% (wt/wt) at 72 h postmortem] on palatability traits of beef (Bos taurus) strip loin steaks. Select (USDA) strip loins were obtained from control (no ZH = 19) and ZH-supplemented carcasses (n = 20). Right and left sides were selected alternatively to serve as a control (no INJ) or CaCl(2)-injected (INJ) and stored at 4°C. Before injecting the subprimals (72 h postmortem), 2 steaks were cut for proximate, sarcomere length, and myofibrillar fragmentation index (MFI) analyses. At 7 d postmortem each strip loin was portioned into steaks, vacuum packaged, and aged for the appropriate period for Warner-Bratzler shear force (WBSF; 7, 14, 21, and 28 d postmortem), trained sensory analysis (14 and 21 d postmortem), purge loss (7 d), and MFI (3, 7, 14, 21, and 28 d postmortem). Results indicated steaks from both ZH supplementation and INJ had reduced WBSF values as days of postmortem aging increased. The WBSF values of ZH steaks were greater (P < 0.05) than no ZH steaks at each postmortem aging period. The INJ steaks had lower WBSF values (P < 0.05) than non-injected steaks. A greater percentage (91 vs. 71%) of steaks had WBSF values < 4.6 kg from steers with no ZH supplementation at 7 d postmortem, but the percentage did not differ (P > 0.05) due to ZH at 14, 21, or 28 d or due to INJ at any aging period. Trained panelists rated tenderness less in ZH steaks than steaks with no ZH at 14 d and 21 d. However, INJ improved (P < 0.05) the tenderness ratings and flavor intensity of the trained panelists, compared with their non-injected cohorts at 21 d. Zilpaterol hydrochloride supplementation reduced (P < 0.05) MFI values, but INJ resulted in greater (P < 0.05) MFI values compared with no INJ. Subprimals from ZH and INJ showed greater purge loss (P < 0.05). Although no interactions were found with ZH and CaCl(2), injecting USDA Select strip loins from ZH-fed cattle can help reduce the normal WBSF variation as it does in steaks from non-ZH-fed cattle.

Shelf life and stability traits of traditionally and modified atmosphere packaged ground beef patties treated with lactic acid bacteria, rosemary oleoresin, or both prior to retail display.

Previous research indicates that lactic acid bacteria (LAB) can inhibit pathogenic bacteria. This research evaluated effects of LAB inclusion on the shelf life of traditionally packaged ground beef patties; as well as the effects and possible interaction of LAB and rosemary oleoresin (RO) on the stability of high oxygen MAP ground beef during display. In both package types, trained and consumer evaluations indicated no effect (P>0.05) of LAB on lean color and off-odor. Display affected trained and consumer sensory evaluations and indicated declined stability over time. Thiobarbituric acid values were lower for traditionally packaged ground beef with LAB (P<0.05) and MAP ground beef with RO or RO and LAB (P<0.05). Overall, LAB had no effect on the shelf life and stability of traditionally or high-oxygen MAP packaged ground beef patties. Therefore, utilization of LAB in ground beef to reduce pathogenic bacteria is viable without alteration of spoilage indicators.

Effect of surface incorporation of broiler litter applied to no-till cotton on runoff quality.

Surface application of broiler litter to no-till cotton could lead to degradation of water quality. Incorporation of broiler litter into the top surface soil (0.05 m) could alleviate this risk. A 2-yr field study was conducted on a silt loam upland soil to determine the effect of incorporation of broiler litter into the soil surface on nutrient and bacterial transport in runoff. The experimental design was a randomized complete block with four treatments and three replications. Treatments were (i) unfertilized control; (ii) surface-appliedbroiler litter at 7.8 Mg ha(-1) without incorporation; (iii) surface-applied broiler litter at 7.8 Mg ha(-1) with immediate incorporation; and (iv) inorganic fertilizer N (urea ammonium nitrate, 32% N) and inorganic fertilizer P (triple superphosphate) at the recommended rate. Phosphorus was surface appliedat 25 kg ha(-1) and N was injected at 101 kg ha(-1) into the soil using a commercial liquid fertilizer applicator. Runoff was collected from small runoff plots (2.4 m by 1.6 m) established at the bottom side of main plots (13.7 m by 6.0 m). Incorporation of broiler litter reduced total N (TN), NO3-N, water soluble P (WSP), and total P (TP) concentrations in runoffby 35, 25, 61, and 64%, respectively, and litter-associated bacteria by two to three orders of magnitude compared with unincorporated treatment. No significant difference in total suspended solids (TSS) in runoffwas obtained between incorporated and unincorporated treatments. Incorporation of broiler litter into the surface soil in the no-till system immediately after application minimized the potential risk for surface nutrient losses and bacteria transport in runoff.

Spoilage and safety characteristics of ground beef packaged in traditional and modified atmosphere packages.

Two separate studies, one with pathogen-inoculated product and one with noninoculated product, were conducted to determine the safety and spoilage characteristics of modified atmosphere packaging (MAP) and traditional packaging of ground beef patties. Ground beef patties were allotted to five packaging treatments (i) control (foam tray with film overwrap; traditional), (ii) high-oxygen MAP (80% 02, 20% CO2), (iii) high-oxygen MAP with added rosemary extract, (iv) low-oxygen carbon monoxide MAP (0.4% CO, 30% CO2, 69.6% N2), and (v) low-oxygen carbon monoxide MAP with added rosemary extract. Beef patties were evaluated for changes over time (0, 1, 3, 5, 7, 14, and 21 days) during lighted display. Results indicated low-oxygen carbon monoxide gas flush had a stabilizing effect on meat color after the formation of carboxymyoglobin and was effective for preventing the development of surface discoloration. Consumers indicated that beef patties packaged in atmospheres containing carbon monoxide were more likely to smell fresh at 7, 14, and 21 days of display, but the majority would probably not consume these products after 14 days of display because of their odor. MAP suppressed the growth of psychrophilic aerobic bacteria when compared with control packages. Generally, control packages had significantly higher total aerobic bacteria and Lactobacillus counts than did modified atmosphere packages. In the inoculated ground beef (approximately 10(5) CFU/g) in MAP, Escherichia coli O157 populations ranged from 4.51 to 4.73 log CFU/g with no differences among the various packages, but the total E. coli O157:H7 in the ground beef in the control packages was significantly higher at 5.61 log CFU/g after 21 days of storage. On days 14 and 21, the total Salmonella in the ground beef in control packages was at 5.29 and 5.27 log CFU/g, respectively, which was significantly higher than counts in the modified atmosphere packages (3.99 to 4.31 log CFU/g on day 14 and 3.76 to 4.02 log CFU/g on day 21). Data from these studies indicate that MAP suppresses pathogen growth compared with controls and that spoilage characteristics developed in MAP packages.

Attentional modulation of visceral and somatic pain.

A better understanding of the cortical processes underlying attentional modulation of visceral and somatic pain in health are essential for interpretation of future imaging studies of hypervigilance towards bodily sensations which is considered to be an aetiologically important factor in the heightened pain reported by patients with irritable bowel syndrome and fibromyalgia. Twelve healthy subjects were recruited for this study. Simultaneous trains of electrical pulses (delivered to either the rectum or lower abdomen) and auditory tones lasting 6 s were delivered to the subjects during a whole-brain functional scan acquisition. Subjects were instructed to attend to the auditory tones (distracter task) or electrical pulses (pain task). Pain intensity ratings were significantly lower during the distraction task compared with the pain task (P < 0.01) in both sensory modalities. The left primary somatosensory cortex increased in activity with increasing pain report, during attention to visceral pain. Bilateral anterior insula (aIns) cortex activity increased with increasing somatic pain report independent of the direction of attention. Conversely, the primary and secondary auditory cortices significantly increased in activation with decreased pain report. These results suggest that pain intensity perception during attentional modulation is reflected in the primary somatosensory cortex (visceral pain) and aIns cortex activity (somatic pain).

Treatments for non-epileptic attack disorder.

BACKGROUND: Psychogenic non-epileptic seizures (NES) have the outward appearance of epilepsy in the absence of physiological or electroencephalographic correlates. Non-epileptic seizures can occur in isolation or in combination with epileptic seizures. The development and maintenance of non-epileptic seizures has been well documented and there is a growing literature on the treatment of NES which includes non-psychological (including anti-anxiety and antidepressant pharmacological treatment) and psychological therapies (including cognitive behavioural therapy (CBT), hypnotherapy and paradoxical therapy). Various treatment methodologies have been tried with variable success. The purpose of this Cochrane review was to establish the evidence base for the treatment of NES. OBJECTIVES: To assess whether treatments for NES result in a reduction in frequency of seizures and/or improvement in quality of life, and whether any treatment is significantly more effective than others. SEARCH STRATEGY: We searched the Cochrane Epilepsy Group's Specialised Register (September 2005), the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library Issue 3, 2005), MEDLINE (1966 to July 2005), and PsycINFO (1806 to July 2005). No language restrictions were imposed. We checked the reference lists of retrieved studies for additional reports of relevant studies SELECTION CRITERIA: Randomised or quasi-randomised studies were included that assessed one or more types of psychological or non-psychological interventions for the treatment of NES. Studies of childhood NES were excluded from our review. DATA COLLECTION AND ANALYSIS: Three review authors independently assessed the trials for inclusion and extracted data. Outcomes included reduction in seizure frequency and improvements in quality of life. MAIN RESULTS: Three small studies met our inclusion criteria and were of poor methodological quality. Two assessed hypnosis and the other paradoxical therapy. There were no detailed reports of improved seizure frequency or quality of life outcomes, and these trials provide no reliable evidence of a beneficial effect of these interventions. AUTHORS' CONCLUSIONS: In view of the methodological limitations and the small number of studies, we have no reliable evidence to support the use of any treatment including hypnosis or paradoxical injunction therapy in the treatment of NES. Randomised studies of these and other interventions are needed.

Effects of feeding whole cottonseed and cottonseed products on performance and carcass characteristics of finishing beef cattle.

Three experiments were conducted to determine the effects of whole cottonseed or cottonseed products on performance and carcass characteristics of beef cattle. In Exp. 1, 120 beef steers (initial BW = 381 +/- 31.7 kg) were fed steam-flaked corn-based finishing diets with 10% (DM basis) basal roughage, and whole cottonseed or individual cottonseed components (cottonseed hulls, meal, and oil). Over the entire feeding period, ADG did not differ (P = 0.95), but DMI increased (P = 0.07) and G:F decreased (P = 0.06) for steers fed the cottonseed diets compared with the control diet. Dressing percent (P = 0.02) and marbling scores (P = 0.02) of carcasses from steers fed the cottonseed diets were less than for steers fed the control diet. In Exp. 2, 150 beef steers (initial BW = 364 +/- 9.9 kg) were used to determine the effects of whole cottonseed or pelleted cottonseed (PCS) on performance and carcass characteristics. Cattle were fed steam-flaked corn-based finishing diets in which whole cottonseed or PCS replaced all of the dietary roughage, supplemental fat, and supplemental natural protein of the control diet. Over the entire feeding period, steers fed the cottonseed diets had lower (P = 0.04) DMI and greater (P < 0.01) G:F than steers fed the control diet. Carcass characteristics did not differ (P = 0.16 to 0.96) among dietary treatments. In Exp. 3, 150 beef heifers (initial BW = 331 +/- 17.1 kg) were used to determine the effects of PCS or delinted, whole cottonseed (DLCS) on performance and carcass characteristics. Heifers were fed rolled corn-based finishing diets in which cottonseed replaced the dietary roughage, supplemental fat, and all or part of the supplemental natural protein of the control diet. Over the entire feeding period, ADG, DMI, and G:F of heifers fed the control diet did not differ (P = 0.19 to 0.80) from those of the cottonseed diets; however, heifers fed the diets containing PCS had greater ADG (P = 0.03) and G:F (P = 0.09) than heifers fed diets containing DLCS. Carcass characteristics of heifers fed the control diet did not differ (P > or = 0.28) from those fed the cottonseed diets. Heifers fed the diets containing PCS had greater (P < or = 0.03) HCW, dressing percent, and LM area than those fed DLCS. Based on our results, whole cottonseed, or products derived from processing whole cottonseed, can replace feedstuffs commonly used in beef cattle finishing diets with no adverse effects on animal performance or carcass characteristics.

A pilot study with long term follow up of hyperbaric oxygen pretreatment in patients with locally advanced breast cancer undergoing neo-adjuvant chemotherapy.

Work in an animal cancer model suggests that pretreatment with hyperbaric oxygen can improve tumor vascularity rendering chemotherapy more effective. Accordingly 32 subjects with locally advanced breast carcinoma (>5cm diameter) entered into a randomized clinical trial where a course was administered of six intravenous pulses of cyclophosphamide 1000mg/m2 i.v., doxorubicin 50mg/m2 i.v. and vincristine 1.5mg/m2 i.v. In the case group this was preceded by ten, once daily, sessions of hyperbaric oxygen therapy (HBO2) administered either at 2.4 or 2.0 atmospheres absolute. Eleven out of 15 subjects tolerated a full course of HBO2 and chemotherapy. All 17 control subjects tolerated a full course of chemotherapy. Tumor extravascular extracellular or edema fluid was reduced after HBO2 but there was no reduction in tumor cell volume and no indication of increased vascularity on MRI. Clinical and pathological responses to chemotherapy were the same in both groups and there was no evidence of neovascularisation. Five year survival in those who tolerated the trial regime was 73% and did not differ between the groups. This mortality was cancer related.

Asphalt volcanism and chemosynthetic life in the Campeche Knolls, Gulf of Mexico.

In the Campeche Knolls, in the southern Gulf of Mexico, lava-like flows of solidified asphalt cover more than 1 square kilometer of the rim of a dissected salt dome at a depth of 3000 meters below sea level. Chemosynthetic tubeworms and bivalves colonize the sea floor near the asphalt, which chilled and contracted after discharge. The site also includes oil seeps, gas hydrate deposits, locally anoxic sediments, and slabs of authigenic carbonate. Asphalt volcanism creates a habitat for chemosynthetic life that may be widespread at great depth in the Gulf of Mexico.

Undernutrition of ewe lambs in utero and in early post-natal life does not affect hypothalamic-pituitary function in adulthood.

The effect of undernutrition in utero, during late gestation (from day 100), and early neonatal life on hypothalamic-pituitary function was investigated in female lambs born to ewes fed rations calculated to provide either 100% (high; H) or 70% (low; L) of the energy requirements to sustain a twin pregnancy. Following parturition in early spring, ewes and lambs were maintained on pasture with sward heights of 6 cm (H) or 4 cm (L) until week 8 of lactation and then sward heights of 5 cm (H) or 3 cm (L) until weaning at week 14. Mean lamb birth weights were 18% lower in L than H animals (P<0.05) and mean liveweights were 23% lower in the L animals (P<0.001) at weaning at 14 weeks of age. Liveweight differences were not significant at, or after, 26 weeks of age. There were no significant differences between pre-pubertal H and L animals, either before (26 weeks) or after ovariectomy (31 weeks), with respect to hypothalamic or pituitary activity, as measured by LH pulse frequency, pulse amplitude or mean plasma LH and FSH concentrations and the responses to GnRH injection as measured by LH peak amplitude, respectively. Similarly there were no differences in any of these variables in pubertal animals at 18 months of age. At 31 weeks of age, H animals had significantly lower pituitary GnRH receptor binding (P<0.01) and lower ERalpha mRNA content (P<0.05) than L lambs. There were no differences with treatment in the abundance of mRNA for LHbeta, FSHbeta or GnRH-receptor at 31 weeks of age or in pubertal animals aged 18 months, when there were no significant differences with treatment in GnRH receptor binding or ERalpha mRNA expression. It is concluded that effects on lifetime reproductive function of female sheep of undernutrition during late gestation and early neonatal life are unlikely to be expressed through permanent changes in hypothalamic-pituitary function and are therefore attributable to effects exerted directly on the ovary.

Plasma selenium level before diagnosis and the risk of prostate cancer development.

PURPOSE: Epidemiological studies and a randomized intervention trial suggest that the risk of prostate cancer may be reduced by selenium intake. We investigated whether plasma selenium level before diagnosis correlated with the risk of later developing prostate cancer. MATERIALS AND METHODS: A case control study was performed on men from the Baltimore Longitudinal Study of Aging registry, including 52 with known prostate cancer and 96 age matched controls with no detectable prostatic disease. Plasma selenium was measured at an average time plus or minus standard deviation of 3.83 +/- 1.85 years before the diagnosis of prostate cancer by graphite furnace atomic absorption spectrophotometry. Adjusted odds ratio and 95% confidence interval were computed with logistic regression. RESULTS: After correcting for years before diagnosis, body mass index, and smoking and alcohol use history, higher selenium was associated with a lower risk of prostate cancer. Compared with the lowest quartile of selenium (range 8.2 to 10.7 microg./dl.), the odds ratios of the second (10.8 to 11.8), third (11.9 to 13.2) and fourth (13.3 to 18.2) quartiles were 0.15 (95% confidence interval 0.05 to 0.50), 0.21 (0.07 to 0.68) and 0.24 (0.08 to 0.77, respectively, p =0.01). Furthermore, plasma selenium decreased significantly with patient age (p <0.001). CONCLUSIONS: Low plasma selenium is associated with a 4 to 5-fold increased risk of prostate cancer. These results support the hypothesis that supplemental selenium may reduce the risk of prostate cancer. Because plasma selenium decreases with patient age, supplementation may be particularly beneficial to older men.