Buckwheat (Fagopyrum esculentum) Hulls Are a Rich Source of Fermentable Dietary Fibre and Bioactive Phytochemicals.

Pseudo-cereals such as buckwheat (Fagopyrum esculentum) are valid candidates to promote diet biodiversity and nutrition security in an era of global climate change. Buckwheat hulls (BHs) are currently an unexplored source of dietary fibre and bioactive phytochemicals. This study assessed the effects of several bioprocessing treatments (using enzymes, yeast, and combinations of both) on BHs' nutrient and phytochemical content, their digestion and metabolism in vitro (using a gastrointestinal digestion model and mixed microbiota from human faeces). The metabolites were measured using targeted LC-MS/MS and GC analysis and 16S rRNA gene sequencing was used to detect the impact on microbiota composition. BHs are rich in insoluble fibre (31.09 ± 0.22% as non-starch polysaccharides), protocatechuic acid (390.71 ± 31.72 mg/kg), and syringaresinol (125.60 ± 6.76 mg/kg). The bioprocessing treatments significantly increased the extractability of gallic acid, vanillic acid, p-hydroxybenzoic acid, syringic acid, vanillin, syringaldehyde, p-coumaric acid, ferulic acid, caffeic acid, and syringaresinol in the alkaline-labile bound form, suggesting the bioaccessibility of these phytochemicals to the colon. Furthermore, one of the treatments, EC_2 treatment, increased significantly the in vitro upper gastrointestinal release of bioactive phytochemicals, especially for protocatechuic acid (p < 0.01). The BH fibre was fermentable, promoting the formation mainly of propionate and, to a lesser extent, butyrate formation. The EM_1 and EC_2 treatments effectively increased the content of insoluble fibre but had no effect on dietary fibre fermentation (p > 0.05). These findings promote the use of buckwheat hulls as a source of dietary fibre and phytochemicals to help meet dietary recommendations and needs.

Homemade Nucleic Acid Preservation Buffer Proves Effective in Preserving the Equine Faecal Microbiota over Time at Ambient Temperatures.

The equine faecal microbiota is often assessed as a proxy of the microbial community in the distal colon, where the microbiome has been linked to states of health and disease in the horse. However, the microbial community structure may change over time if samples are not adequately preserved. This study stored equine faecal samples from n = 10 horses in four preservation treatments at room temperature for up to 150 h and assessed the resulting impact on microbial diversity and the differential abundance of taxa. Treatments included "COLD" (samples packaged with a cool pack), "CLX" (2% chlorhexidine digluconate solution), "NAP" (nucleic acid preservation buffer), and "FTA" (Whatman FTA™ cards). The samples were assessed using 16S rRNA gene sequencing after storage for 0, 24, 72, and 150 h at room temperature under the different treatments. The results showed effective preservation of diversity and community structure with NAP buffer but lower diversity (p = 0.001) and the under-representation of Fibrobacterota in the FTA card samples. The NAP treatment inhibited the overgrowth of bloom taxa that occurred by 72 h at room temperature. The COLD, CLX, and NAP treatments were effective in preserving the faecal microbiota for up to 24 h at room temperature, and the CLX and NAP treatments improved the yield of Patescibacteria and Fibrobacterota in some cases. The cold and CLX treatments were ineffective in preventing community shifts that occurred by 72 h at room temperature. These findings demonstrate the suitability of the COLD, NAP, and CLX treatments for the room temperature storage of equine faeces for up to 24 h and of NAP buffer for up to 150 h prior to processing.

Confidence does not mediate a relationship between owner experience and likelihood of using weight management approaches for native ponies.

Native ponies are at increased risk of obesity and metabolic perturbations, such as insulin dysregulation (ID), a key risk factor for endocrinopathic laminitis. Management and feeding practices can be adapted to maintain healthy body condition and support metabolic health, but owners may inadvertently provide their ponies with inappropriate management leading to obesity and exacerbating risk of metabolic disease. Adoption of preventative weight management approaches (WMAs), including regular monitoring of body condition, providing appropriate preserved forage, promoting seasonal weight loss, and using exercise accordingly, are key in supporting native ponies' metabolic health. The factors influencing the adoption of WMAs, such as owners' experience and confidence, require exploration. The aim of the current study was to understand factors influencing owners' likelihood to undertake certain WMAs, to develop our understanding of suitable intervention targets. A total of 571 responses to an online cross-sectional questionnaire were analysed. Mediation analysis revealed that whilst long term (≥20 years) experience caring for native ponies was associated with owners increased, self-reported confidence in identifying disease and managing their native ponies, this did not translate to an increased likelihood of implementing WMAs. Conversely, respondents who managed ponies with dietary requirements related to obesity, laminitis, or equine metabolic syndrome were more likely to use WMAs related to feeding, seasonal weight management and exercise. Owner confidence was assessed and rejected as a mediator of the relationship between experience and WMA use. These results highlight the need for further work that elucidates the pathways leading owners to undertake action against obesity without the need for ponies to develop overt disease, as well as suggesting a need for long term managers of native ponies to update management practices with preventative care as the focus.

Green Technologies for Persimmon By-Products Revalorisation as Sustainable Sources of Dietary Fibre and Antioxidants for Functional Beverages Development.

The use of green technologies such as ultrasound and natural deep eutectic solvents (NADES) for revalorisation of food and agricultural by-products represents a sustainable way to tackle waste and promote a healthier environment while delivering much-needed functional food ingredients for an increasingly unhealthy population. The processing of persimmon (Diospyros kaki Thunb.) generates large amounts of by-products rich in fibre-bound bioactive phytochemicals. This paper assessed the extractability of bioactive compounds through NADES and the functional properties of the persimmon polysaccharide-rich by-products to evaluate their suitability to be used as functional ingredients in commercial beverages. Although higher amounts of carotenoids and polyphenols were extracted after eutectic treatment vs. conventional extraction (p < 0.05), the fibre-bound bioactives remained abundant (p < 0.001) in the resulting persimmon pulp by-product (PPBP) and persimmon pulp dietary fibre (PPDF), showing also a strong antioxidant activity (DPPH•, ABTS•+ assays) and an improved digestibility and fibre fermentability. The main structural components of PPBP and PPDF are cellulose, hemicellulose and pectin. PPDF-added dairy-based drink showed more than 50% of preference over the control among panellists and similar acceptability scores to the commercial ones. Persimmon pulp by-products represent sustainable source of dietary fibre and bioactives and are suitable candidates to develop functional ingredients for food industry applications.

Habitual consumption of high-fibre bread fortified with bean hulls increased plasma indole-3-propionic concentration and decreased putrescine and deoxycholic acid faecal concentrations in healthy volunteers.

Only 6 to 8 % of the UK adults meet the daily recommendation for dietary fibre. Fava bean processing lead to vast amounts of high-fibre by-products such as hulls. Bean hull fortified bread was formulated to increase and diversify dietary fibre while reducing waste. This study assessed the bean hull: suitability as a source of dietary fibre; the systemic and microbial metabolism of its components and postprandial events following bean hull bread rolls. Nine healthy participants (53·9 ± 16·7 years) were recruited for a randomised controlled crossover study attending two 3 days intervention sessions, involving the consumption of two bread rolls per day (control or bean hull rolls). Blood and faecal samples were collected before and after each session and analysed for systemic and microbial metabolites of bread roll components using targeted LC-MS/MS and GC analysis. Satiety, gut hormones, glucose, insulin and gastric emptying biomarkers were also measured. Two bean hull rolls provided over 85 % of the daily recommendation for dietary fibre; but despite being a rich source of plant metabolites (P = 0·04 v. control bread), these had poor systemic bioavailability. Consumption of bean hull rolls for 3 days significantly increased plasma concentration of indole-3-propionic acid (P = 0·009) and decreased faecal concentration of putrescine (P = 0·035) and deoxycholic acid (P = 0·046). However, it had no effect on postprandial plasma gut hormones, bacterial composition and faecal short chain fatty acids amount. Therefore, bean hulls require further processing to improve their bioactives systemic availability and fibre fermentation.

Hemp and buckwheat are valuable sources of dietary amino acids, beneficially modulating gastrointestinal hormones and promoting satiety in healthy volunteers.

PURPOSE: This study evaluated the postprandial effects following consumption of buckwheat, fava bean, pea, hemp and lupin compared to meat (beef); focussing on biomarkers of satiety, gut hormones, aminoacids and plant metabolites bioavailability and metabolism. METHODS: Ten subjects (n = 3 men; n = 7 women; 42 ± 11.8 years of age; BMI 26 ± 5.8 kg/m2) participated in six 1-day independent acute interventions, each meal containing 30 g of protein from buckwheat, fava bean, pea, hemp, lupin and meat (beef). Blood samples were collected during 24-h and VAS questionnaires over 5-h. RESULTS: Volunteers consumed significantly higher amounts of most amino acids from the meat meal, and with few exceptions, postprandial composition of plasma amino acids was not significantly different after consuming the plant-based meals. Buckwheat meal was the most satious (300 min hunger scores, p < 0.05).Significant increase in GLP-1 plasma (AUC, iAUC p = 0.01) found after hemp compared with the other plant-based meals. Decreased plasma ghrelin concentrations (iAUC p < 0.05) found on plant (hemp) vs. meat meal. Several plasma metabolites after hemp meal consumption were associated with hormone trends (partial least squares-discriminant analysis (PLS-DA): 4-hydroxyphenylpyruvic acid, indole 3-pyruvic acid, 5-hydoxytryptophan, genistein and biochanin A with GLP-1, PYY and insulin; 3-hydroxymandelic acid and luteolidin with GLP-1 and ghrelin and 4-hydroxymandelic acid, benzoic acid and secoisolariciresinol with insulin and ghrelin. Plasma branched-chain amino acids (BCAAs), (iAUC, p < 0.001); and phenylalanine and tyrosine (iAUC, p < 0.05) were lower after buckwheat comparison with meat meal. CONCLUSION: Plants are valuable sources of amino acids which are promoting satiety. The impact of hemp and buckwheat on GLP-1 and, respectively, BCAAs should be explored further as could be relevant for aid and prevention of chronic diseases such as type 2 diabetes. Study registered with clinicaltrial.gov on 12th July 2013, study ID number: NCT01898351.

The Human Aspect of Horse Care: How the COVID-19 Pandemic Impacted the Wellbeing of Equestrian Industry Stakeholders.

During the lockdown phase of the COVID-19 pandemic, equestrian stakeholders faced a dilemma whereby they were required to balance caring for the welfare of horses with adapting to the restrictions imposed to protect public health. The present study investigated the impact of the pandemic on the wellbeing of a sample of industry stakeholders, including horse owners, equine veterinarians, farriers and welfare centre managers (n = 26) using a qualitative methodology. Findings from the interviews indicated that the mental health and wellbeing of veterinarians and horse owners was negatively affected by pandemic-related obstacles to communication and limitations to horse-owner interactions. However, this study also identified several positive outcomes for wellbeing during lockdown resulting from pro-social activities that were engaged with by horse owners to overcome social isolation, the separation of the community and loneliness. These findings provide accounts of ways in which those caring for horses might be challenged during national emergency scenarios, pointing to areas that would benefit from future mental health and wellbeing interventions.

COVID-19 impacts equine welfare: Policy implications for laminitis and obesity.

The COVID-19 pandemic continues to impact human health and welfare on a global level. In March 2020, stringent national restrictions were enforced in the UK to protect public health and slow the spread of the SARS-CoV-2 virus. Restrictions were likely to have resulted in collateral consequences for the health and welfare of horses and ponies, especially those at risk of obesity and laminitis and this issue warranted more detailed exploration. The current study utilised qualitative methodology to investigate the implications of COVID-19 related policies upon equine management and welfare with a focus on horses and ponies at risk of laminitis and obesity. Twenty-four interviews with five sub-groups of key equestrian welfare stakeholders in the UK were conducted between May and July of 2020 to understand the challenges facing equine welfare in the context of laminitis and obesity susceptible animals. Thematic analysis revealed lockdown-associated factors with the potential to compromise welfare of horses and ponies at risk of obesity and laminitis. These included: disparate information and guidance, difficulties enacting public health measures in yard environments, and horses having reduced exercise during the pandemic. Positive examples of clear and consistent information sharing by farriers were reported to have improved horse owner understanding of routine hoof care during lockdown. Analysis suggested that the recommendations for supporting the management-based needs of horses under reduced supervision were not clearly defined, or were not sufficiently disseminated, across the equine industry. These findings support the development of guidelines specific to the care of horses and ponies at risk of obesity and laminitis through collaborative input from veterinary and welfare experts, to reduce the negative impacts of future lockdown events in the UK.

Sapogenol is a Major Microbial Metabolite in Human Plasma Associated with High Protein Soy-Based Diets: The Relevance for Functional Food Formulations.

Legumes are a source of health-promoting macro- and micronutrients, but also contain numerous phytochemicals with useful biological activities, an example of which are saponins. Epidemiological studies suggest that saponins may play a role in protection from cancer and benefit human health by lowering cholesterol. Therefore, they could represent good candidates for specialised functional foods. Following the consumption of a soya-rich high-protein weight-loss diet (SOYA HP WL), the concentrations of Soyasaponin I (SSI) and soyasapogenol B (SSB) were determined in faecal samples from human volunteers (n = 10) and found to be between 1.4 and 17.5 mg per 100 g fresh faecal sample. SSB was the major metabolite identified in volunteers' plasma (n = 10) after consumption of the soya test meal (SOYA MEAL); the postprandial (3 h after meal) plasma concentration for SSB varied between 48.5 ng/mL to 103.2 ng/mL. The metabolism of SSI by the gut microbiota (in vitro) was also confirmed. This study shows that the main systemic metabolites of soyasaponin are absorbed from the gut and that they are bioavailable in plasma predominantly as conjugates of sapogenol. The metabolism and bioavailability of biologically active molecules represent key information necessary for the efficient development of functional foods.

Rapid method for quantification of anthocyanidins and anthocyanins in human biological samples.

Using simple solvent extraction and enzymatic hydrolysis, a rapid LC-MS/MS method for quantification of free and conjugated forms of anthocyanidins and anthocyanins in plasma and urine samples was developed and validated. A mixed enzymatic treatment containing ß-glucuronidase (100 U mL-1) and sulfatase (2.5 U mL-1) for 5 min (37 °C; pH 6) was optimal condition for deconjugation of anthocyanidins and anthocyanins in urine and plasma samples. The LC-MS/MS allowed quantifying thirteen different anthocyanidins and anthocyanins simultaneously. The developed LC-MS/MS method was precise and accurate over multiple days and nominal concentrations. The stability assessment study confirmed that the long-term storage and/or periodic use of plasma and urine samples might have a considerable impact on the stability of some anthocyanidins. The method was successfully applied to measure anthocyanidins and anthocyanins in plasma and urine samples following consumption of acute blueberry test meals.

Availability and dose response of phytophenols from a wheat bran rich cereal product in healthy human volunteers.

SCOPE: Phytophenols present in cereals are metabolised to compounds that could be partly responsible for the reduced risk of chronic diseases and all-cause mortality associated with fibre-rich diets. The bioavailability, form and in vivo concentrations of these metabolites require to be established. MATERIALS AND METHODS: Eight healthy volunteers consumed a test meal containing a recommended dose (40 g) and high dose (120 g) of ready-to-eat wheat bran cereal and the systemic and colonic metabolites determined quantitatively by LC-MS. CONCLUSION: Analysis of the systemic metabolomes demonstrated that a wide range of phytophenols were absorbed/excreted (43 metabolites) within 5 h of consumption. These included 16 of the 21 major parent compounds identified in the intervention product and several of these were also found to be significantly increased in the colon. Not all of the metabolites were increased with the higher dose, suggesting some limitation in absorption due to intrinsic factors and/or the food matrix. Many compounds identified (e.g. ferulic acid and major metabolites) exhibit anti-inflammatory activity and impact on redox pathways. The combination of postprandial absorption and delivery to the colon, as well as hepatic recycling of the metabolites at these concentrations, is likely to be beneficial to both systemic and gut health.

Nutritional and Phytochemical Content of High-Protein Crops.

Sustainable sources of high-protein plants could help meet future protein requirements. Buckwheat, green pea, fava bean, hemp, and lupin were analyzed by proximate analysis and inductively coupled plasma mass spectrometry to determine their macro- and micronutrient contents, and liquid chromatography-mass spectrometry was used to elucidate the phytochemical profiles. The protein contents ranged from 20 to 43% (w/w), and all samples were found to be rich in insoluble fiber: 9-25% (w/w). The selected crops had a favorable micronutrient profile, with phosphorus levels ranging from 2.22 ± 0.05 to 9.72 ± 0.41 g kg-1, while iron levels ranged from 20.23 ± 0.86 to 69.57 ± 7.43 mg kg-1. The crops contained substantial amounts of phytophenolic compounds. In particular, buckwheat was a rich source of pelargonidin (748.17 ± 75.55 mg kg-1), epicatechin (184.1 ± 33.2 mg kg-1), quercetin (35.66 ± 2.22 mg kg-1), caffeic acid (41.74 ± 22.54 mg kg-1), and 3-hydroxyphenylacetic acid (63.64 ± 36.16 mg kg-1); hemp contained p-coumaric acid (84.02 ± 8.10 mg kg-1), cyanidin (58.43 ± 21.01 mg kg-1), protocatechualdehyde (34.77 ± 5.15 mg kg-1), and gentisic acid (31.20 ± 1.67 mg kg-1); and fava bean was the richest source of ferulic acid (229.51 ± 36.58 mg kg-1) and its 5-5' (39.99 ± 1.10 mg kg-1) and 8-5 dimers (58.17 ± 6.68 mg kg-1). Demonstrating that these crops are rich sources of protein, fiber, and phytochemicals could encourage higher consumption and utilization of them as healthy and sustainable ingredients in the food and drink industry.

Anti- and pro-oxidative effect of fresh and freeze-dried vegetables during storage of mayonnaise.

Mayonnaise was supplemented with vegetables (5 % w/w) and the effect of storage time at 4 °C on the oxidative stability of the dispersed phase was investigated. Results indicated that mayonnaise is prone to lipid oxidation during storage under refrigerator conditions. The type of vegetable used for mayonnaise reformulation was critical in inhibiting oxidation and followed the order beetroot > carrot ≈ onion with respect to antioxidant capacity. Broccoli induced a pro-oxidant effect and the rate of oxidation by the end of the storage period was 42 times higher compared with the control. The addition of beetroot, either fresh or freeze-dried, improved the oxidative stability of mayonnaise significantly. The process of freeze-drying affected adversely the ability of vegetables to decrease oil oxidation of the emulsions. This may reflect loss of important natural antioxidants during the drying procedure.

Comparative study of the functional properties of lupin, green pea, fava bean, hemp, and buckwheat flours as affected by pH.

The demand for products of high nutritional value from sustainable sources is growing rapidly in the global food market. In this study, the effect of pH on the functional properties of lupin, green pea, fava bean, hemp, and buckwheat flours was investigated and compared with wheat flour. Functional properties included solubility, emulsifying and foaming properties, gelling ability, and water holding capacity (WHC). All flours had minimal solubility at pH 4 and their corresponding values increased with increasing pH. Emulsifying properties were improved at pH 10 for all samples and emulsion stability showed a similar trend. Increasing pH in the range 4-10 enhanced the foaming properties of the flours, particularly buckwheat and hemp. Wheat, green pea, buckwheat, and fava bean were more capable of forming firm gels compared with lupin and hemp, as indicated by least gelling concentrations (LGCs). The ranking of the water binding properties of the different types of flours were lupin>hemp>fava bean>buckwheat>green pea>wheat. Results indicate that underutilized flours from sustainable plant sources could be exploited by the food industry as functional food ingredients or as replacements of wheat flour for various food applications. Depending on the application, flour functionality may be effectively tailored by pH adjustment.

Appetite control and biomarkers of satiety with vegetarian (soy) and meat-based high-protein diets for weight loss in obese men: a randomized crossover trial.

BACKGROUND: There is limited evidence with regard to the effect of different sources of protein on appetite during weight loss. Vegetarian and meat-based high-protein diets may have contrasting effects on appetite and biomarkers of protein-induced satiety. OBJECTIVE: The aim was to assess appetite response to meat or vegetarian high-protein weight-loss (HPWL) diets in obese men to monitor plasma amino acid profile and gut peptide response as potential satiety biomarkers. DESIGN: Twenty obese [body mass index (in kg/m²): 34.8] men participated in a dietary intervention study. After 3 d of a maintenance diet, they were provided in a crossover design with either a vegetarian HPWL (Soy-HPWL) or a meat-based HPWL (Meat-HPWL) diet for 2 wk. Both diets comprised 30% protein, 30% fat, and 40% carbohydrate, provided to measured resting metabolic rate. Body weight and the motivation to eat were measured daily. Plasma satiety biomarkers were collected during a test-meal challenge (5 h) at the end of each diet period. RESULTS: Over the 2 wk, subjects lost, on average, 2.41 and 2.27 kg with consumption of the Soy- and Meat-HPWL diets, respectively [P = 0.352; SE of the difference (SED): 0.1]. ANOVA confirmed that subjectively rated hunger (P = 0.569; SED: 3.8), fullness (P = 0.404; SED: 4.1), desire to eat (P = 0.356; SED: 3.7), preservation of lean body mass (P = 0.334; SED: 0.2), and loss of percentage fat mass (P = 0.179; SED: 0.2) did not differ between the 2 HPWL diets. There were differences in absolute concentrations of ghrelin and peptide YY between the 2 HPWL diets, although the response as net area under the curve was not different. CONCLUSIONS: Appetite control and weight loss were similar for both HPWL diets. Gut hormone profile was similar between the diets, which suggests that vegetarian diets can be as effective as meat-based diets for appetite control during weight loss.

Total and individual carotenoids and phenolic acids content in fresh, refrigerated and processed spinach (Spinacia oleracea L.).

The carotenoid and phenolic acid contents in fresh, stored and processed (blanched, frozen and boiled) spinach were comparatively determined by spectrophotometric and HPLC analyses. The major carotenoids identified after HPLC analysis in saponified samples were lutein (37-53µg/kg), ß-carotene (18-31µg/kg), violaxanthin (9-23µg/kg) and neoxanthin (10-22µg/kg). These carotenoids were all affected by storage and/or heating. The content of carotenoids was best preserved after storage for one day at 4°C. The total phenolic content in the fresh spinach was 2088mg GAE/kg FW. After LC-MS analysis three phenolic acids were identified and quantified. These being ortho-coumaric acid (28-60mg/kg FW), ferulic acid (10-35mg/kg) and para-coumaric acid (1-30mg/kg) depending on the sample type. After storage of spinach at different temperatures (4°C or -18°C) the amount of total phenolic compounds decreased by around 20%, while the amount of individual phenolic acids increased by four times on average.

A novel antioxidant phenyl disaccharide from Populus tremula knotwood.

The complete characterization of two compounds obtained from the acetone extract of Populus tremula knotwood has been was achieved using LC-DAD-MS, MS/MS, IR and NMR. The new compounds were unequivocally identified as a mixture of the ester isomers of the (E) and (Z) p-coumarate of 1-O-rutinose. The isomers showed the capacity to inhibit lipid peroxidation induced by tert-butylhydroperoxide and to trap peroxyl radicals, as determined by a chemiluminescence method. These new phenyl disaccharides also showed a significant ORAC (oxygen radical absorbance capacity) value, i.e. 11.7 microM TE (Trolox Equivalents).