Redox balance and immunity of piglets pre- and post-E. coli challenge after treatment with hemp or fish oil, and vitamin E.

This study investigated the influence of polyunsaturated fatty acid composition and vitamin E supplementation on oxidative status and immune responses in weanling piglets pre- and post-E. coli challenge. Suckling piglets (n = 24) were randomly selected from two litters for an oral supplementation (1 mL/day) with fish oil or hemp oil and vitamin E supplementation (60 mg natural vitamin E/mL oil) from day 10 to 28 of age. At day 29 and 30 of age, each piglet was orally inoculated with 6.7 × 108 and 3.96 × 108 CFU of F4 and F18 E. coli, respectively. Blood was sampled from all piglets on day 28 before E. coli challenge and on day 35 of age to investigate immunological and oxidative stress markers in plasma. One week after weaning and exposure to E. coli, a general reduction in the α-tocopherol concentration and activity of GPX1 was obtained. Vitamin E supplementation lowered the extent of lipid peroxidation and improved the antioxidative status and immune responses after E. coli challenge. Hemp oil had the greatest effect on antioxidant enzyme activity. Provision of hemp oil and vitamin E to suckling piglets may reduce the incidence of post-weaning diarrhea.

A high fat to vitamin E ratio in the feed protects and improves uptake of the natural form of vitamin E in postweaning calves.

In postweaning calves, it is a challenge to maintain the plasma vitamin E level at or above the recommended level (3 µg/mL), which is linked to a good immune response. It has been unclear until now why the provision of solid feed with concentrations below 200 mg/kg feed of vitamin E is ineffective in maintaining the plasma vitamin E level of calves above the recommended plasma level postweaning. The present study was conducted to investigate if a high fat to vitamin E ratio in the concentrate could protect and improve the delivery of the natural form of vitamin E (RRR-α-tocopherol) to calves postweaning. Thirty calves were included in the experiment from 2 weeks preweaning until 2 weeks postweaning (Weeks -2, -1, 0 [weaning], 1, and 2 relative to weaning) and fed one of three concentrates in which lecithin mixture provided the fat supplement: control (77 mg/kg of vitamin E and 4.9% DM of crude fat; CONT), medium level of vitamin E supplemented (147 mg/kg of vitamin E and 7.7% DM of crude fat; MedVE) or high level of vitamin E supplemented (238 mg/kg of vitamin E and 12.4% DM of fat; HiVE). Thus, there was a comparable ratio of fat to vitamin E (520-630) in the three concentrates. During the 2 weeks postweaning, final body weight (92 ± 2 kg), average daily gain (917 ± 51 g/day) and concentrate intake (2.2 ± 0.09 kg/day; mean of treatment ± standard error) were unaffected by treatment and the interaction between treatment and week. There was an interaction between treatment and week for vitamin E intake pre- (p < 0.001) and postweaning (p < 0.001). There was an interaction between treatment and week (p < 0.001) for plasma vitamin E level postweaning, and it was 2.5, 3.1, and 3.8 µg/mL in CONT, MedVE, and HiVE, respectively, at Week 1 postweaning. In addition, plasma vitamin E levels at Week 2 postweaning were 2.6, 3.6 and 4.8 µg/mL in CONT, MidVE and HiVE respectively. The results show that 147 mg/kg of lecithin-protected vitamin E in the concentrate is needed to secure a plasma vitamin E level well above the recommended level. In addition, lecithin-protected vitamin E elevated the plasma level of triglycerides and nonesterified fatty acids.

Formation of RRR-α-tocopherol in rumen and intestinal digestibility of tocopherols in dairy cows.

Tocopherol sources in diets are often a combination of all-rac-α-tocopheryl acetate (synthetic α-tocopherol) from vitamin supplements and natural tocopherols and 2R-(4'R, 8'R)-5,7,8-trimethyltocotrienol (α-tocotrienols) from the feed sources. Synthetic α-tocopherol consists of 8 different stereoisomers including 2R-(4'R, 8'R)-5,7,8-trimethyltocol (RRR-α-tocopherol), 2R-(4'S, 8'R)-5,7,8-trimethyltocol (RSR-α-tocopherol), 2R-(4'R, 8'S)-5,7,8-trimethyltocol (RRS-α-tocopherol), 2R-(4'S, 8'S)-5,7,8-trimethyltocol (RSS-α-tocopherol), 2S-(4'S, 8'S)-5,7,8-trimethyltocol (SSS-α-tocopherol), 2S-(4'R, 8'S)-5,7,8-trimethyltocol (SRS-α-tocopherol), 2S-(4'S, 8'R)-5,7,8-trimethyltocol (SSR-α-tocopherol), and 2S-(4'R, 8'R)-5,7,8-trimethyltocol (SRR-α-tocopherol). The pre-absorption metabolism of tocopherols and tocotrienols in ruminants differs from monogastric animals due to the extensive microbial fermentation in the anaerobic rumen. The current study investigated the impact of toasting and decortication of oats on metabolism in the digestive tract (synthesis, digestion), and intestinal digestibility of tocopherols in dairy cows by using 4 ruminal and intestinal cannulated Danish Holstein cows in a 4 × 4 Latin square design for 4 periods. Cows were fed a total mixed ration ad libitum containing different forms of oats: whole oat, decorticated oat, toasted oat, and decorticated toasted oat, all rolled before mixed ration. Overall means across 4 treatments were statistically analyzed, testing whether overall means were different from zero. Decortication or toasting did not affect the balance or digestibility of α-tocopherols in rumen. Average across treatments showed the ruminal degradation of synthetic α-tocopherol (279 mg/d, P = 0.02; P-value shows that average across treatments is different from zero), synthetic 2R-α-tocopherol (133 mg/d, P < 0.01; summation of RRS-, RSR- and RSS-α-tocopherol), and 2S-α-tocopherol (190 mg/d; P < 0.01, summation of SSS-, SRS-, SSR, and SRR-α-tocopherol), while RRR-α-tocopherol was formed in the rumen (221 mg/d, P = 0.10). The average across treatments showed that small intestinal digestibility of tocopherols ranked in the following order: α-tocotrienol > natural α-tocopherol > synthetic α-tocopherols > 2R-(4'R, 8'R)-,7,8-dimethyltocol (γ-tocopherol). The average across treatments for small intestinal and feed-ileum digestibility ranked in the following order: RRR-α-tocopherol > synthetic 2R-α-tocopherol > 2S-α-tocopherol. Results showed the first evidence for RRR-α-tocopherol formation under anaerobic conditions in the rumen. In addition, synthetic α-tocopherol stereoisomers, γ-tocopherol and α-tocotrienol were degraded in the rumen. There was a discrimination against absorption of synthetic 2R- and 2S-α-tocopherol in the small intestine.

The effect of dietary omega-3 fatty acid supplementation on fetal growth, piglet birth weight and plasma fatty acid concentrations, using docosahexaenoic acid in early gestation in sows.

The objective of the study was to test the effect of the omega-3 fatty acid docosahexaenoic acid (DHA) on fetal and placental development as well as the birth weight of piglets. A total of 238 multiparous sows were allocated to either a control diet group or a DHA diet group with an omega-6 to omega-3 ratio of 9.8 and 2.4, respectively, from mating to day 43 of gestation. A blood sample was collected and back fat thickness was measured prior to mating, on days 14, 42 and 112 of gestation. On day 43 of gestation, 14 sows were slaughtered and measurements of fetuses and placentas were taken. Piglets in some litters were weighed individually at farrowing. Dietary treatment did not affect fetal characteristics and back fat thickness (P > 0.05). Dietary treatment increased the plasma concentrations of total omega-3 fatty acids in sows (P < 0.05). Sows fed the DHA diet had a shorter gestation length compared to the control sows (P < 0.05), but the number of born piglets was not affected (P > 0.05). The average piglet birth weight and the within-litter variation in birthweight were unaffected by dietary DHA (P > 0.05), however, sows fed DHA diet had fewer piglets under 800 g at birth compared to control sows (P < 0.05). In conclusion, addition of DHA decreased the dietary ratio of omega-6 to omega-3 fatty acids, increased plasma n-3 fatty acid concentrations in sows and decreased the number of piglets weighing under 800 g at birth.

Supplementation of palmitoleic acid improved piglet growth and reduced body temperature drop upon cold exposure.

Piglet survival is a major challenge in the first few days postpartum and interventions during this period may improve survival and growth. This study investigated the effects of palmitoleic acid (C16:1n-7; PA) supplementation on growth performance, body temperature, fatty acid (FA), and energy metabolism in milk-replacer-fed piglets. Forty-eight piglets were stratified by body weight and randomly assigned to one of four dietary treatments (0%, 1%, 2%, and 3% PA supplementation as a percent of milk replacer) and given the diet through an orogastric tube. They were fed dietary treatments every 2 h for 4 d in the first week postpartum and all were sacrificed at the end of the experiment. The piglets were weighed daily, and half in each dietary treatment group, the same piglets each day, were exposed daily to a lower temperature for 2 h. Plasma samples were collected immediately before sacrifice for analyses of FA and other plasma metabolites. The weight of organs and empty body weight were determined after sacrifice. Liver and semimembranosus muscle tissue samples were collected and analyzed for FA content. Contents of C16:1n-7 and C18:1n-7 in both plasma and liver (P < 0.001), and C16:1n-7 in semimembranosus muscle (P < 0.001) increased linearly as PA supplementation increased. Most plasma FA levels (except C16:1n-7, C16:1n-9, and C22:5n-3) were lower in piglets exposed to lower temperatures than those that were not. Plasma glucose, triglycerides, and lactate dehydrogenase levels increased linearly with PA supplementation (P < 0.001). Piglets' average daily gain, liver glycogen pool, liver weight, and gallbladder weight increased linearly (P < 0.05, P < 0.01, P < 0.05, and P < 0.001, respectively), but lung weight, liver nitrogen content, and body temperature drop decreased linearly (P < 0.01, P < 0.001, and P < 0.05, respectively) with PA supplementation. Piglets exposed to low temperature had greater liver nitrogen (P < 0.05) and lactate dehydrogenase (P < 0.001) contents but had lower liver weight (P < 0.01) and plasma lactate concentration (P < 0.05) than those that were not. In conclusion, this study demonstrated the importance of PA on the growth performance of the piglets by increasing their average daily gain and decreasing a drop in body temperature upon cold exposure, most likely due to a modified energy metabolism.

Reducing piglet mortality in the early days after birth is a significant challenge in the modern pig industry. The focus on achieving larger litter sizes has had a negative impact on piglets' birth weight and their intake of colostrum. Additionally, piglets are born without easily oxidizable brown adipose tissue and have limited body reserves, making them more vulnerable to death due to their lower capacity for thermogenesis. Therefore, it is important to explore dietary strategies that can enhance piglets' thermogenesis capacity. In this study, the role of palmitoleic acid supplementation was investigated in a dose-response design to determine its impact on growth performance, fatty acid composition, and energy metabolism of milk-replacer-fed piglets during their first week of life. The results revealed a linear increase in the average daily gain of the piglets, liver weight, and liver glycogen content with increasing palmitoleic acid supplementation. Moreover, increased palmitoleic acid supplementation was associated with a drop in body temperature when piglets were exposed to a lower temperature during the experimental period. Altogether, the study indicated that palmitoleic acid has a sparing effect on glycogen reserves and that a greater proportion of energy utilized by the piglets to maintain their body temperature was derived from the oxidation of fatty acids. The results indicated a promising approach to improve piglet survival and growth through dietary modifications of fatty acids in the diet.

Monitoring of environmental DNA from nonindigenous species of algae, dinoflagellates and animals in the North East Atlantic.

Monitoring the distribution of marine nonindigenous species is a challenging task. To support this monitoring, we developed and validated the specificity of 12 primer-probe assays for detection of environmental DNA (eDNA) from marine species, all nonindigenous to Europe. The species include sturgeons, a Pacific red algae, oyster thief, a freshwater hydroid from the Black Sea, Chinese mitten crab, Pacific oyster, warty comb jelly, sand gaper, round goby, pink salmon, rainbow trout and North American mud crab. We tested all assays in the laboratory, on DNA extracted from both the target and non-target species to ensure that they only amplified DNA from the intended species. Subsequently, all assays were used to analyse water samples collected at 16 different harbours across two different seasons during 2017. We also included six previously published assays targeting eDNA from goldfish, European carp, two species of dinoflagellates of the genera Karenia and Prorocentrum, two species of the heterokont flagellate genus Pseudochattonella. Conventional monitoring was carried out alongside eDNA sampling but with only one sampling event over the one year. Because eDNA was relatively fast and easy to collect compared to conventional sampling, we sampled eDNA twice during 2017, which showed seasonal changes in the distribution of nonindigenous species. Comparing eDNA levels with salinity gradients did not show any correlation. A significant correlation was observed between number of species detected with conventional monitoring methods and number of species found using eDNA at each location. This supports the use of eDNA for surveillance of the distribution of marine nonindigenous species, where the speed and relative easy sampling in the field combined with fast molecular analysis may provide advantages compared to conventional monitoring methods. Prior validation of assays increases taxonomic precision, and laboratorial setup facilitates analysis of multiple samples simultaneously. The specific eDNA assays presented here can be implemented directly in monitoring programmes across Europe and potentially worldwide to infer a more precise picture of the dynamics in the distribution of marine nonindigenous species.

Biorefinery of Green Biomass─How to Extract and Evaluate High Quality Leaf Protein for Food?

There is a growing need for protein for both feed and food in order to meet future demands. It is imperative to explore and utilize novel protein sources such as protein from leafy plant material, which contains high amounts of the enzyme ribulose-1,5-biphosphate carboxylase/oxygenase (RuBisCo). Leafy crops such as grasses and legumes can in humid climate produce high protein yields in a sustainable way when compared with many traditional seed protein crops. Despite this, very little RuBisCo is utilized for foods because proteins in the leaf material has a low accessibility to monogastrics. In order to utilize the leaf protein for food purposes, the protein needs to be extracted from the fiber rich leaf matrix. This conversion of green biomass to valuable products has been labeled green biorefinery. The green biorefinery may be tailored to produce different products, but in this Review, the focus is on production of food-grade protein. The existing knowledge on the extraction, purification, and concentration of protein from green biomass is reviewed. Additionally, the quality and potential application of the leaf protein in food products and side streams from the green biorefinery will be discussed along with possible uses of side streams from the protein production.

Absorption of α-tocopheryl acetate is limited in mink kits (Mustela vison) during weaning.

Bioavailability of α-tocopherol varies with source, dose and duration of supplementation. The effect of source and dose of α-tocopherol on response of α-tocopherol stereoisomers in plasma and tissues of mink kits during the weaning period was studied. Twelve mink kits were euthanised in CO2 at the beginning of the experiment, and 156 mink kits (12 replicates per treatment group) were randomly assigned to thirteen treatment groups: no added α-tocopherol in the feed (0 dose) or four different doses (50, 75, 100 and 150 mg/kg of diet) of RRR-α-tocopherol (ALC), RRR-α-tocopheryl acetate (ACT) or all-rac-α-tocopheryl acetate (SYN). Six mink kits per treatment group were euthanised 3 weeks after initiation of the experiment, and the remaining six were euthanised 6 weeks after initiation of the experiment. The RRR-α-tocopherol content in plasma, liver, heart and lungs was affected by interaction between source and dose (P < 0.01 for all). The highest RRR-α-tocopherol content in plasma (13.6 µg/ml; LS-means for source across dose and week), liver (13.6 µg/mg), heart (7.6 µg/mg) and lungs (9.8 µg/mg) was observed in mink kits fed ALC. The RRR-α-tocopherol content in plasma and tissues depended on source and dose interaction and increased linearly with supplementation. In conclusion, the interaction between source and dose reveals a limitation in hydrolysis of ester bond in α-tocopheryl acetate in mink kits around weaning as the likely causative explanation for the higher response of ALC at the highest doses. Thus, considerable attention has to be paid to the source of α-tocopherol during weaning of mink kits fed a high dose of α-tocopherol.

Sentinel Coronavirus environmental monitoring can contribute to detecting asymptomatic SARS-CoV-2 virus spreaders and can verify effectiveness of workplace COVID-19 controls.

Detecting all asymptomatic or presymptomatic COVID-19 virus spreaders at a workplace requires daily testing of employees by RT-PCR, which is not practical. Over a two week period, 9 Europe and USA workplace locations were chosen to test employees for SARS-CoV-2 infection (841 tests) and high-frequency-touch point environmental surfaces (5,500 tests) for Coronavirus by RT-PCR. Of the 9 locations, 3 had one or more employees infected with SARS-CoV-2 during the two week study period. None of the employees who tested positive had symptoms at the time of testing and none developed symptoms during subsequent 14 day quarantine. Locations with significant prevalence of Coronavirus contaminated environmental surfaces were 10 times more likely to have a positive employees than locations with no or very few environmental surfaces positive for Coronavirus. Break room chairs, workbenches, and break room door handles were the most frequently contaminated environmental surfaces. Surface Coronavirus RNA was detected at very low concentrations (RT-PCR 34 to 38 Cq). These results suggest that Coronavirus environmental monitoring may have potential to predict presence of asymptotic spreaders and to validate and verify COVID-19 control strategies on a regular basis.

Changes in long-chain fatty acid composition of milk fat globule membrane and expression of mammary lipogenic genes in dairy cows fed sunflower seeds and rumen-protected choline.

The aim of this experiment was to investigate the effect of dietary supplementation of crushed high oleic sunflower seeds (HOS) and rumen-protected choline (RPC) on the fatty acid (FA) profile of phospholipids and sphingomyelin and mammary transcription of genes that are important for milk fat synthesis and de novo synthesis of sphingolipids. Twenty-four cows were divided into four groups that either received an unsupplemented diet (Control), the Control diet supplemented with 50 g RPC per day, a diet supplemented with HOS at 10% of dry matter, or RPC and HOS in combination (RPC + HOS). RPC supplementation had no effect on the FA composition of milk or sphingomyelin. Cows receiving RPC and RPC + HOS had increased incorporation of C22:5 (n-3) into phospholipids. Milk FA proportion of C18:0 and C18:1 isomers was increased in cows receiving HOS (HOS and RPC + HOS). Sphingomyelin proportion of C22:0 was increased in cows receiving HOS and RPC + HOS, at the expense of C23:0. HOS supplementation further increased the proportion of unsaturated fatty acids (UFA) in milk phospholipids. HOS supplementation increased mammary transcription of UDP-glucose ceramide glycosyltransferase (UGCG), sterol response element-binding protein cleavage-activating protein (SCAP) and peroxisome proliferation-activated receptor Gamma subunit C 1b (PPARGC1b), and reduced transcription of insulin induced gene 1 (INSIG1) and fatty acid-binding protein 3 (FABP3). Dietary supplementation of RPC increased mammary transcription of fatty acid desaturase 1 (FADS1) and longevity assurance gene 2 (LASS2), and reduced transcription of sphingomyelin synthase (SGMS). The results show that the FA profile of milk phospholipids is sensitive to dietary lipid supplementation and, to a minor degree, RPC supplementation. Furthermore, transcription of genes that are important for milk fat synthesis and sphingolipids synthesis is affected by dietary supplementation of RPC and HOS.

Non-enzymatic transformations of dietary 2-hydroxyalkenyl and aromatic glucosinolates in the stomach of monogastrics.

Monogastric animals exhibit different biological responses to structurally diverse glucosinolates and their transformation products, depending on the dietary levels. The transformations of 2-hydroxyalkenyl and aromatic glucosinolates were examined in vitro under gastric conditions, ex vivo in ligated porcine stomachs and in vivo in a rat model. Intact glucosinolates were almost completely transformed in vitro within 1 h at pH 3 (73-88%) and at pH 5 (97-100%) upon addition of Fe2+ ranging from two-fold molar excess. Glucosinolate transformations reached 78-99% when incubated ex vivo in ligated porcine stomachs. Rat in vivo feeding trials showed major reductions (81-84%) in the intact glucosinolate contents upon passage through the gastrointestinal (GI) tract. Non-enzymatic transformations of glucosinolates occur in the stomach, where pH and the level of Fe2+ are primary determinants. This is the first study to show a complex formation between iron-progoitrin and iron-sinalbin, facilitating the transformation into nitriles and thionamides.

Distribution of α-tocopherol stereoisomers in mink (Mustela vison) organs varies with the amount of all-rac-α-tocopheryl acetate in the diet.

Synthetic α-tocopherol has eight isomeric configurations including four 2R (RSS, RRS, RSR, RRR) and four 2S (SRR, SSR, SRS, SSS). Only the RRR stereoisomer is naturally synthesised by plants. A ratio of 1·36:1 in biopotency of RRR-α-tocopheryl acetate to all-rac-α-tocopheryl acetate is generally accepted; however, studies indicate that neither biopotency of α-tocopherol stereoisomers nor bioavailability between them is constant, but depend on dose, time, animal species and organs. A total of forty growing young male mink were, after weaning, assigned one of the following treatments for 90 d: no α-tocopherol in diet (ALFA_0), 40 mg/kg RRR-α-tocopheryl acetate (NAT_40), 40 mg/kg all-rac-α-tocopheryl acetate (SYN_40) and 80 mg/kg feed all-rac-α-tocopheryl acetate (SYN_80). Mink were euthanised in CO2 and blood was collected by heart puncture. Mink were pelted and liver, heart, lungs, brain and abdominal fat were collected for α-tocopherol stereoisomer analysis. The proportion of RRR-α-tocopherol decreased in all organs and plasma with increasing amount of synthetic α-tocopherol stereoisomers in the diet (P≤0·05), whereas the proportion of all synthetic α-tocopherol stereoisomers increased with increasing amount of synthetic α-tocopherol stereoisomers in the diet (P≤0·05). The proportion of α-tocopherol stereoisomers in plasma, brain, heart, lungs and abdominal fat showed the following order: RRR>RRS, RSR, RSS>Σ2S, regardless of α-tocopherol supplement. The liver had the highest proportion of Σ2S stereoisomers, and lowest proportion of RRR-α-tocopherol. In conclusion, distribution of α-tocopherol stereoisomers differs with dose and form of α-tocopherol supplementation. The results did also reveal the liver's role as the major organ for accumulation of Σ2S α-tocopherol stereoisomers.

RRR-α-Tocopherol Is the Predominant Stereoisomer of α-Tocopherol in Human Milk.

BACKGROUND: The naturally occurring α-tocopherol (α-T) stereoisomer, RRR-α-tocopherol (RRR-α-T), is known to be more bioactive than all-rac-α-tocopherol (all-rac-α-T), a synthetic racemic mixture of 8 stereoisomers. There is widespread use of all-rac-α-T in maternal supplements. OBJECTIVE: The aim of the study was to thoroughly describe the α-T stereoisomer profile of human milk. METHODS: We measured the α-T stereoisomer profile in milk from 2 cohorts of women: a cohort of 121 women who provided milk on days 30 and 60 of lactation (study 1) and a separate cohort of 51 women who provided milk on days 10, 21, 71, and 120 of lactation (study 2). RESULTS: RRR-α-T was the predominant stereoisomer (P < 0.0001) in all samples in both studies despite a large intrasubject range in total α-T (0.7-22 µg/mL). On average, RRR-α-T comprised 73-76% of total α-T, but average values for the synthetic stereoisomers were RRS, 8-14%; RSR, 6-8%; RSS, 5-6%; and the sum of 2S stereoisomers (Σ2S), 3-5%. Despite the predominance of RRR-α-T, the sum of the synthetic stereoisomers comprised as much as 48% of total α-T. We calculated the ratio of RRR to the sum of the synthetic 2R (RRS + RSR + RSS) stereoisomers (s2R) to assess the degree to which RRR is favored in milk. Consistent with discrimination among 2R stereoisomers in mammary tissue, RRR/s2R values ranged from 2.8 to 3.6, as opposed to the expected ratio of 0.33 if there was no discrimination. However, the RRR to s2R ratio did not correlate with milk α-T concentration, but both components of the ratio did. CONCLUSIONS: RRR-α-T is the predominant stereoisomer in human milk, concentrations of synthetic 2R stereoisomers were notable, and the relation between milk total α-T and stereoisomer profile is complex. Due to the wide range found in milk α-T stereoisomer profile, investigation into its impact on α-T status and functional outcomes in breastfed infants is warranted.

White clover fractions as protein source for monogastrics: dry matter digestibility and protein digestibility-corrected amino acid scores.

BACKGROUND: The present study aimed to evaluate the use of white clover as an alternative protein source for monogastrics. White clover plant and leaves were processed using a screw-press resulting in a solid pulp and a juice from which protein was acid-precipitated. The chemical composition of all fractions was determined and digestibility of dry matter (DM) and protein was assessed in an experiment with growing rats. RESULTS: Protein concentrates were produced with crude protein (CP) contents of 451 g kg-1 and 530 g kg-1 DM for white clover plant and leaves, respectively, and a pulp with CP contents of 313 and 374 g kg-1 DM from plant and leaves, respectively. The amino acid composition ranged from 4.72 to 6.49 g per 16 g of nitrogen (N) for lysine, 1.82-2.6 g per 16 g N for methionine and cysteine, and 3.66-5.24 g per 16 g N for threonine. True faecal digestibility of protein varied from 0.81 to 0.88, whereas DM digestibility was in the range 0.72-0.80. Methionine and cysteine were found to be limiting in all fractions, regardless of the reference group used. CONCLUSION: A high digestibility of white clover protein was found irrespective of the physical fractionation. Together with a well-balanced amino acid composition, this makes white clover a promising protein source for monogastrics. © 2017 Society of Chemical Industry.

Effects of vitamin D and its metabolites on cell viability and Staphylococcus aureus invasion into bovine mammary epithelial cells.

Vitamin D has been found have various biological effects that may be potent in preventing bovine mastitis. Two forms of vitamin D, vitamin D2 (D2) and vitamin D3 (D3), can be hydroxylated to functional metabolites in cattle. The objectives of the present study were to investigate the potential of vitamin D compounds for controlling bovine mastitis using in vitro cell models, and to compare the differences between D2 and D3 compounds. Results showed that D2 compounds have comparable effects to their D3 analogues on inhibiting MAC-T cell viability in vitro. S. aureus growth was inhibited by high concentrations of D2, D3, 25(OH)D2 and 25(OH)D3. 25(OH)D2 and 25(OH)D3 induced CYP24A1 expression but reduced VDR mRNA expression, whereas the expression of CYP27B1, occludin, and E-cadherin did not change. Additionally, the induction of CYP24A1 expression by 25(OH)D3 was higher than that of 25(OH)D2, which may contribute to their differences in inhibiting cell viability. S. aureus invaded into MAC-T cells and universally inhibited gene expressions. Pre-treat MAC-T cells with 25(OH)D2 reduced S. aureus adhesion while pre-treatment with 25(OH)D3 inhibited S. aureus invasion, but neither of the compounds attenuated the S. aureus-induced gene expression reduction. In conclusion, the present study shows that D2 compounds have comparable effects on inhibiting cell viability and S. aureus invasion to their D3 analogues in vitro, suggesting that D2 and its metabolites have potential in controlling bovine mastitis.

Supplementation with RRR- or all-rac-α-Tocopherol Differentially Affects the α-Tocopherol Stereoisomer Profile in the Milk and Plasma of Lactating Women.

Background: The naturally occurring α-tocopherol stereoisomer RRR-α-tocopherol is known to be more bioactive than synthetic α-tocopherol (all-rac-α-tocopherol). However, the influence of this difference on the α-tocopherol stereoisomer profile of human milk is not understood.Objective: We investigated whether supplemental RRR-α-tocopherol or all-rac-α-tocopherol differentially affected the distribution of α-tocopherol stereoisomers in milk and plasma from lactating women.Methods: Eighty-nine lactating women aged 19-40 y and with a body mass index (in kg/m2) ≤30 were randomly assigned at 4-6 wk postpartum to receive a daily supplement containing 45.5 mg all-rac-α-tocopherol acetate (ARAC), 22.8 mg all-rac-α-tocopherol acetate + 20.1 mg RRR-α-tocopherol (MIX), or 40.2 mg RRR-α-tocopherol (RRR). Milk and plasma were analyzed for α-tocopherol structural isomers and α-tocopherol stereoisomers at baseline and after 6 wk supplementation with the use of chiral HPLC.Results: There were no significant treatment group or time-dependent changes in milk or plasma α, γ, or δ-tocopherol. RRR-α-tocopherol was the most abundant stereoisomer in both milk and plasma in each group. Supplementation changed both milk and plasma percentage RRR-α-tocopherol (RRR > MIX > ARAC) (P < 0.05) and percentage non-RRR-α-tocopherol (ARAC > MIX > RRR) (P < 0.05). In the RRR group, percentage RRR-α-tocopherol increased in milk (mean ± SEM: 78% ± 2.3% compared with 82% ± 1.7%) (P < 0.05) and plasma (mean ± SEM: 77% ± 1.8% compared with 87% ± 1%) (P < 0.05). In contrast, the percentage RRR-α-tocopherol decreased in the MIX and ARAC groups (MIX, P < 0.05; ARAC, P < 0.0001), and percentage non-RRR-α-tocopherol stereoisomers increased (MIX, P < 0.05; ARAC, P < 0.0001) commensurate with an accumulation of 2S-α-tocopherol stereoisomers (P < 0.05) in both milk and plasma. Milk and plasma RRR-α-tocopherol was positively correlated at baseline (r = 0.67; P < 0.0001) and 6 wk (r = 0.80; P < 0.0001).Conclusion: The α-tocopherol supplementation strategy differentially affected the α-tocopherol milk and plasma stereoisomer profile in lactating women. RRR-α-tocopherol increased milk and plasma percentage RRR-α-tocopherol, whereas all-rac-α-tocopherol acetate reduced these percentages. Because RRR-α-tocopherol is the most bioactive stereoisomer, investigating the impact of supplement-driven changes in the milk α-tocopherol stereoisomer profile on the α-tocopherol status of breastfed infants is warranted.

The naturally occurring α-tocopherol stereoisomer RRR-α-tocopherol is predominant in the human infant brain.

α-Tocopherol is the principal source of vitamin E, an essential nutrient that plays a crucial role in maintaining healthy brain function. Infant formula is routinely supplemented with synthetic α-tocopherol, a racaemic mixture of eight stereoisomers with less bioactivity than the natural stereoisomer RRR-α-tocopherol. α-Tocopherol stereoisomer profiles have not been previously reported in the human brain. In the present study, we analysed total α-tocopherol and α-tocopherol stereoisomers in the frontal cortex (FC), hippocampus (HPC) and visual cortex (VC) of infants (n 36) who died of sudden infant death syndrome or other conditions. RRR-α-tocopherol was the predominant stereoisomer in all brain regions (P<0·0001) and samples, despite a large intra-decedent range in total α-tocopherol (5-17 µg/g). Mean RRR-α-tocopherol concentrations in FC, HPC and VC were 10·5, 6·8 and 5·5 µg/g, respectively. In contrast, mean levels of the synthetic stereoisomers were RRS, 1-1·5; RSR, 0·8-1·0; RSS, 0·7-0·9; and Σ2S 0·2-0·3 µg/g. Samples from all but two decedents contained measurable levels of the synthetic stereoisomers, but the intra-decedent variation was large. The ratio of RRR:the sum of the synthetic 2R stereoisomers (RRS+RSR+RSS) averaged 2·5, 2·3 and 2·4 in FC, HPC and VC, respectively, and ranged from 1 to at least 4·7, indicating that infant brain discriminates against synthetic 2R stereoisomers in favour of RRR. These findings reveal that RRR-α-tocopherol is the predominant stereoisomer in infant brain. These data also indicate that the infant brain discriminates against the synthetic 2R stereoisomers, but is unable to do so completely. On the basis of these findings, investigation into the impact of α-tocopherol stereoisomers on neurodevelopment is warranted.

Interactions between retinol, α-tocopherol and cholecalciferol need consideration in diets for farmed mink (Mustela vison).

A sufficient but balanced vitamin supplementation is a prerequisite for a satisfactory growth pattern and an effective immune system in mink and all other species. The fat-soluble vitamins are very sensitive to over- or under-supply because they interact with each other with respect to dose-response and chemical form. The purpose of the present study was to investigate the effect of increasing the amount of retinol in combination with RRR-α-tocopherol or all-rac-α-tocopherol in the feed given to growing mink on their retinol, cholecalciferol and α-tocopherol concentrations in plasma and selected organs. The results showed that the mink met their retinol requirements from the basal diet, but there were no negative effects of supplying various amounts of retinol on their plasma α-tocopherol concentrations. On the other hand, the study showed that the cholecalciferol status in plasma, assessed as the 25-hydroxycholecalciferol concentration, was low when retinol was supplemented in the feed at high levels. In addition, supplementation with RRR-α-tocopherol in the feed negatively affected the plasma concentration of 25-hydroxycholecalciferol compared with supplementation with all-rac-α-tocopherol. In general, female mink had higher concentrations of fat-soluble vitamins in plasma than male mink.

25-hydroxyvitamin D circulates in different fractions of calf plasma if the parent compound is vitamin D2 or vitamin D3, respectively.

Vitamin D has become one of the most discussed nutrients in human nutrition, which has led to an increased interest in milk as a vitamin D source. Problems related to fortifying milk with synthetic vitamin D can be avoided by securing a high content of natural vitamin D in the milk by supplying dairy cows with sufficient vitamin D. However, choosing the most efficient route and form of supplementation requires insight into how different vitamin D metabolites are transported in the body of cattle. There are two forms of vitamin D: vitamin D2 (D2) and vitamin D3 (D3). Vitamin D2 originates from fungi on roughage. Vitamin D3 originates either from endogenous synthesis in the skin or from feed supplements. Vitamin D2 is chemically different from, and less physiologically active than, D3. Endogenous and dietary D3 is chemically similar but dietary D3 is toxic, whereas endogenous D3 appears well regulated in the body.

Cheddar Cheese Ripening Affects Plasma Nonesterified Fatty Acid and Serum Insulin Concentrations in Growing Pigs.

BACKGROUND: Meta-analyses of observational studies found cheese consumption to be inversely associated with risk of type 2 diabetes and metabolic syndrome. This may be attributed to the bioactive compounds produced during cheese ripening. OBJECTIVE: The objective of this study was to investigate by means of a porcine model how cheeses with different ripening times affect blood glucose, insulin, and lipid concentrations and fecal-fat excretion. METHODS: A parallel-arm randomized intervention study with 36 Landrace × Yorkshire × Duroc crossbred 3-mo-old female pigs was conducted. The pigs were fed a 21-d butter-rich run-in diet (143 g of butter/kg diet), followed by a 14-d intervention with 1 of 3 isocaloric diets: 4-mo ripened cheddar (4-MRC) diet, 14-mo ripened cheddar (14-MRC) diet, or 24-mo ripened cheddar (24-MRC) diet (350 g of cheese/kg diet). Serum cholesterol, triglycerides, HDL cholesterol, LDL cholesterol, and insulin; plasma nonesterified fatty acids (NEFAs) and glucose; fecal-fat excretion; and body weight were measured. RESULTS: Plasma NEFAs were lower in the 24-MRC (201 ± 26 µEq/L) and in the 14-MRC (171 ± 19 µEq/L) diet groups than in the 4-MRC diet group (260 ± 27 µEq/L; P = 0.044 and P = 0.001). Serum insulin was lower in the 24-MRC diet group (1.04 ± 0.09 mmol/L) than in the 4-MRC diet group (1.44 ± 0.14 mmol/L; P = 0.002), but intermediate and not different from either in the 14-MRC diet group (1.25 ± 0.11 mmol/L). Likewise, homeostasis model assessment of insulin resistance was lower in the 24-MRC diet group (0.030 ± 0.003) than in the 4-MRC diet group (0.041 ± 0.005; P < 0.01), but intermediate and not different from either in the 14-MRC group (0.036 ± 0.004). CONCLUSIONS: Intake of long-term ripened cheddar improved indicators of insulin sensitivity in growing pigs compared with short-term ripened cheddar. This may also be important for human health.