Some Insect Species Are Good-Quality Protein Sources for Children and Adults: Digestible Indispensable Amino Acid Score (DIAAS) Determined in Growing Pigs.

BACKGROUND: Insect species are suitable for farming as "mini livestock" for human and animal consumption. It is important to assess the protein quality of relevant species to understand the potential of these novel protein sources in future sustainable food systems. OBJECTIVES: We aimed to determine the protein quality of 5 insect species-lesser mealworm (LMW), yellow mealworm, house cricket (HC), banded cricket (BC), and black soldier fly-using the digestible indispensable amino acid score (DIAAS) in a pig model. METHODS: Five diets were formulated to contain 10% insect crude protein (CP). A nitrogen (N)-free diet was included to estimate endogenous losses. In a 6 × 6 Latin square design, 6 ileal cannulated crossbred [Duroc × (Danish Landrace × Yorkshire)] male pigs with an initial body weight of 35 ± 2 kg were fed the 6 diets. Each diet was fed for 1 wk over 6 wk. Ileal digesta were collected for 8 h on days 5 and 7 each week. Analyzed CP, amino acid (AA) contents, and calculated values of standardized ileal digestibility for CP and AAs were used to assess the DIAAS of each insect. RESULTS: The DIAAS for young children aged 6 mo-3 y and for older children, adolescents, and adults identified sulfur AAs (cysteine + methionine) as the first limiting AA in all 4 species of cricket and mealworm. For young children, both cricket species had DIAASs > 75 and for older children, adolescents, and adults both cricket species and LMW had DIAASs > 75. CONCLUSIONS: Both cricket species (HC and BC) are classified as good-quality protein sources for young children aged 6 mo-3 y and for older children, adolescents, and adults. One mealworm species, LMW, is a good-quality protein source for older children, adolescents, and adults.

Some Insect Species Are Good-Quality Protein Sources for Children and Adults: Digestible Indispensable Amino Acid Score (DIAAS) Determined in Growing Pigs.

BACKGROUND: Insect species are suitable for farming as "mini livestock" for human and animal consumption. It is important to assess the protein quality of relevant species to understand the potential of these novel protein sources in future sustainable food systems. OBJECTIVES: We aimed to determine the protein quality of 5 insect species-lesser mealworm (LMW), yellow mealworm, house cricket (HC), banded cricket (BC), and black soldier fly-using the digestible indispensable amino acid score (DIAAS) in a pig model. METHODS: Five diets were formulated to contain 10% insect crude protein (CP). A nitrogen (N)-free diet was included to estimate endogenous losses. In a 6 × 6 Latin square design, 6 ileal cannulated crossbred [Duroc × (Danish Landrace × Yorkshire)] male pigs with an initial body weight of 35 ± 2 kg were fed the 6 diets. Each diet was fed for 1 wk over 6 wk. Ileal digesta were collected for 8 h on days 5 and 7 each week. Analyzed CP, amino acid (AA) contents, and calculated values of standardized ileal digestibility for CP and AAs were used to assess the DIAAS of each insect. RESULTS: The DIAAS for young children aged 6 mo-3 y and for older children, adolescents, and adults identified sulfur AAs (cysteine + methionine) as the first limiting AA in all 4 species of cricket and mealworm. For young children, both cricket species had DIAASs > 75 and for older children, adolescents, and adults both cricket species and LMW had DIAASs > 75. CONCLUSIONS: Both cricket species (HC and BC) are classified as good-quality protein sources for young children aged 6 mo-3 y and for older children, adolescents, and adults. One mealworm species, LMW, is a good-quality protein source for older children, adolescents, and adults.

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.

Rye Bran Modified with Cell Wall-Degrading Enzymes Influences the Kinetics of Plant Lignans but Not of Enterolignans in Multicatheterized Pigs.

Background: Whole-grain intake is associated with a lower risk of chronic Western-style diseases, possibly brought about by the high concentration of phytochemicals, among them plant lignans (PLs), in the grains.Objective: We studied whether treatment of rye bran with cell wall-degrading enzymes changed the solubility and kinetics of PLs in multicatheterized pigs.Methods: Ten female Duroc × Danish Landrace × Yorkshire pigs (60.3 ± 2.3 kg at surgery) fitted with permanent catheters were included in an incomplete crossover study. The pigs were fed 2 experimental diets for 1-7 d. The diets were rich in PLs and based on nontreated lignan-rich [LR; lignan concentration: 20.2 mg dry matter (DM)/kg] or enzymatically treated lignan-rich (ENZLR; lignan concentration: 27.8 mg DM/kg) rye bran. Plasma concentrations of PLs and enterolignans were quantified with the use of targeted LC-tandem mass spectrometry. Data were log transformed and analyzed with mixed-effects, 1-compartment, and asymptotic regression models.Results: The availability of PLs was 38% greater in ENZLR than in LR, and the soluble fraction of PLs was 49% in ENZLR compared with 35% in LR diets. PLs appeared in the circulation 30 min after intake of both the ENZLR and LR diets. Postprandially, consumption of ENZLR resulted in a 4-times-greater (P < 0.0001) plasma PL concentration compared with LR. The area under the curve (AUC) measured 0-360 min after ENZLR intake was ∼2 times higher than after LR intake. A 1-compartment model could describe the postprandial increase in plasma concentration after ENZLR intake, whereas an asymptotic regression model described the plasma concentrations after LR intake. Despite increased available and soluble PLs, ENZLR did not increase plasma enterolignans.Conclusion: The modification of rye bran with cell wall-degrading enzymes resulted in significantly greater plasma concentrations of PLs and the 4-h AUC, particularly syringaresinol, in multicatheterized pigs.

Effect of Antibiotics and Diet on Enterolactone Concentration and Metabolome Studied by Targeted and Nontargeted LC-MS Metabolomics.

High plant lignan intake is associated with a number of health benefits, possibly induced by the lignan metabolite enterolactone (ENL). The gut microbiota plays a crucial role in converting dietary lignans into ENL, and epidemiological studies have shown that use of antibiotics is associated with lower levels of ENL. Here we investigate the link between antibiotic use and lignan metabolism in pigs using LC-MS/MS. The effect of lignan intake and antibiotic use on the gut microbial community and the pig metabolome is studied by 16S rRNA sequencing and nontargeted LC-MS. Treatment with antibiotics resulted in substantially lower concentrations of ENL compared with concentrations detected in untreated animals, whereas the plasma concentrations of plant lignans were unchanged. Both diet and antibiotic treatment affected the clustering of urinary metabolites and significantly altered the proportions of taxa in the gut microbiota. Diet, but not antibiotic treatment, affected the plasma lipid profile, and a lower concentration of LDL cholesterol was observed in the pigs fed a high lignan diet. This study provides solid support for the associations between ENL concentrations and use of antibiotics found in humans and indicates that the lower ENL concentration may be a consequence of the ecological changes in the microbiota.

Ferulic acid dehydrodimer and dehydrotrimer profiles of distiller's dried grains with solubles from different cereal species.

Ferulic acid dehydrodimers (DFA) and dehydrotrimers (TriFA) ester-linked to plant cell wall polymers may cross-link not only cell wall polysaccharides but also other cell wall components including proteins and lignin, thus enhancing the rigidity and potentially affecting the enzymatic degradation of the plant cell wall. Corn, wheat, and mixed-cereal distiller's dried grains with solubles (DDGS) were investigated for composition of DFAs and TriFAs by reversed phase high-performance liquid chromatography with ultraviolet detection. Corn DDGS contained 5.3 and 5.9 times higher contents of total DFAs than wheat and mixed-cereal DDGS, respectively. Furthermore, the contents of total TriFAs were 5.7 and 6.3 times higher in corn DDGS than in wheat and mixed-cereal DDGS, respectively. In addition, both corn grains and corresponding DDGS had similar profiles of individual DFAs and TriFAs, indicating that ferulic acid cross-links in the corn cell wall are presumably not modified during fermentation and DDGS processing.

Simultaneous pharmacokinetic modeling of alkylresorcinols and their main metabolites indicates dual absorption mechanisms and enterohepatic elimination in humans.

BACKGROUND: Alkylresorcinols have proven to be useful biomarkers of whole-grain wheat and rye intake in many nutritional studies. To improve their utility, more knowledge regarding the fate of alkylresorcinols and their metabolites after consumption is needed. OBJECTIVE: The objective of this study was to develop a combined pharmacokinetic model for plasma concentrations of alkylresorcinols and their 2 major metabolites, 3,5-dihydroxybenzoic acid (DHBA) and 3-(3,5-dihydroxyphenyl)-propanoic acid (DHPPA). METHODS: The model was established by using plasma samples collected from 3 women and 2 men after a single dose (120 g) of rye bran and validated against fasting plasma concentrations from 8 women and 7 men with controlled rye bran intake (23, 45, or 90 g/d). Alkylresorcinols in the lymph and plasma of a pig fed a single alkylresorcinol dose (1.3 mmol) were quantified to assess absorption. Human ileostomal effluent and pig bile after high and low alkylresorcinol doses were analyzed to evaluate biliary alkylresorcinol metabolite excretion. RESULTS: The model contained 2 absorption compartments: 1 that transferred alkylresorcinols directly to the systematic circulation and 1 in which a proportion of absorbed alkylresorcinols was metabolized before reaching the systemic circulation. Plasma concentrations of alkylresorcinols and their metabolites depended on absorption and formation, respectively, and the mean ± SEM terminal elimination half-life of alkylresorcinols (1.9 ± 0.59 h), DHPPA (1.5 ± 0.26 h), and DHBA (1.3 ± 0.22 h) did not differ. The model accurately predicted alkylresorcinol and DHBA concentrations after repeated alkylresorcinol intake but DHPPA concentration was overpredicted, possibly because of poorly modeled enterohepatic circulation. During the 8 h following administration, <2% of the alkylresorcinol dose was recovered in the lymph. DHPPA was identified in both human ileostomal effluent and pig bile, indicating availability of DHPPA for absorption and enterohepatic circulation. CONCLUSION: Intact alkylresorcinols have advantages over DHBA and DHPPA as plasma biomarkers for whole-grain wheat and rye intake because of lower susceptibility to factors other than alkylresorcinol intake.

Diets high in resistant starch and arabinoxylan modulate digestion processes and SCFA pool size in the large intestine and faecal microbial composition in pigs.

The effects of a high level of dietary fibre (DF) either as arabinoxylan (AX) or resistant starch (RS) on digestion processes, SCFA concentration and pool size in various intestinal segments and on the microbial composition in the faeces were studied in a model experiment with pigs. A total of thirty female pigs (body weight 63.1 (sem 4.4) kg) were fed a low-DF, high-fat Western-style control diet (WSD), an AX-rich diet (AXD) or a RS-rich diet (RSD) for 3 weeks. Diet significantly affected the digestibility of DM, protein, fat, NSP and NSP components, and the arabinose:xylose ratio, as well as the disappearance of NSP and AX in the large intestine. RS was mainly digested in the caecum. AX was digested at a slower rate than RS. The digesta from AXD-fed pigs passed from the ileum to the distal colon more than twice as fast as those from WSD-fed pigs, with those from RSD-fed pigs being intermediate (P< 0.001). AXD feeding resulted in a higher number of Faecalibacterium prausnitzii, Roseburia intestinalis, Blautia coccoides-Eubacterium rectale, Bifidobacterium spp. and Lactobacillus spp. in the faeces sampled at week 3 of the experimental period (P< 0.05). In the caecum, proximal and mid colon, AXD feeding resulted in a 3- to 5-fold higher pool size of butyrate compared with WSD feeding, with the RSD being intermediate (P <0.001). In conclusion, the RSD and AXD differently affected digestion processes compared with the WSD, and the AXD most efficiently shifted the microbial composition towards butyrogenic species in the faeces and increased the large-intestinal butyrate pool size.

Similar metabolic responses in pigs and humans to breads with different contents and compositions of dietary fibers: a metabolomics study.

BACKGROUND: In nutritional studies, pigs are often used as models for humans because of nutritional and physiologic similarities. However, evidence supporting similar metabolic responses to nutritional interventions is lacking. OBJECTIVE: The objective was to establish whether pigs and humans respond similarly to a nutritional intervention. Using metabolomics, we compared the acute metabolic response to 4 test breads between conventional pigs (growing) and adult human subjects (with the metabolic syndrome). DESIGN: Six catheterized pigs and 15 human subjects were tested in a randomized crossover design with 4 breads: white-wheat bread low in dietary fiber, rye bread with whole-rye kernels, and 2 white-wheat breads supplemented with either wheat arabinoxylan or oat ß-glucan. Blood samples drawn -15, 30, and 120 min postprandially were analyzed by untargeted liquid chromatography-mass spectrometry metabolomics. RESULTS: We found that the postprandial responses, as reflected in blood metabolomes, are similar in pigs and humans. Twenty-one of 26 identified metabolites that were found to be different between the species were qualitatively similar in response to the test breads, despite different basal metabolome concentrations in the plasma of pigs and humans. Humans had higher contents of phosphatidylcholines, oleic acid, and carnitine in plasma, possibly reflecting a higher intake of meats and fats. In pigs, betaine, choline, creatinine, tryptophan, and phenylalanine were higher, probably because of the higher doses of bread provided to the pigs (per kg body weight) and/or because of their growing status. Acute metabolic differences in these metabolites induced by the breads were, however, comparable between the 2 species. CONCLUSION: Our results indicate that pigs are a suitable model for human metabolic studies in food research. The human trial was registered at clinicaltrials.gov as NCT01316354. The animal experiment was conducted according to a license obtained by the Danish Animal Experiments Inspectorate, Ministry of Food, Agriculture and Fisheries, Danish Veterinary and Food Administration.

Concentrated arabinoxylan but not concentrated ß-glucan in wheat bread has similar effects on postprandial insulin as whole-grain rye in porto-arterial catheterized pigs.

The acute glycemic effects of concentrated dietary fibers (DF) versus whole-grain rye were studied in porto-arterial catheterized pigs. Two white wheat breads with wheat arabinoxylan (AX) or oat ß-glucan (BG), two rye breads with intact rye kernels (RK) or milled rye (GR), and a low DF white wheat bread were fed to six pigs in a randomized crossover design. Blood profiles were collected for 4 h after feeding. Glucose absorption was reduced in pigs fed the AX bread at 60 min postprandial (3.1 mmol/min for AX compared to 9.4 mmol/min for WF, P = 0.02) and insulin secretion was lowered at 30 min postprandial for AX and GR (74.4 and 129 pmol/min for AX and GR, respectively, compared to 738 pmol/min for WF, P < 0.04). In conclusion, the GR and AX breads were most effective in improving insulin economy, suggesting that arabinoxylan from wheat and rye induces similar outcomes in the metabolic response.

Duodenal application of Li+ in a submaximal therapeutic dose inhibits exocrine pancreatic secretion and modulates gastro-duodenal myoelectrical activity in a conscious pig model.

This study tested whether duodenal application of lithium inhibits gastroduodenal motility, and whether it suppresses secretion from the exocrine pancreas. Five suckling pigs, 16-18 days old, were surgically fitted with 3 serosal electrodes on the wall of the gastric antrum and the duodenum for electromyography of smooth muscles, and with a pancreatic duct catheter and a duodenal T-cannula for collection and re-entrant flow of pancreatic juice. After the recovery period, on alternative days, each animal was tested once with an intraduodenal infusion of Li+ (100 mmol·L(-1) C3H5LiO3, 10 mL·kg(-1)·h(-1)) for 1 h, and once with an intraduodenal infusion of NaCl (154 mM, 10 ml·kg(-1)·h(-1)), also for 1 h, with the first treatment, i.e., Li+ or NaCl, randomly assigned. Individual pigs served as their own controls, with data recorded prior to a treatment being used as the baseline. Li+ increased the duration of quiescence (P < 0.05) and activity phase (P < 0.05) in the antrum, thus increasing (P < 0.05) the duration of antral myoelectrical cycles. Li+ shortened (P < 0.05) phase I, but it did not affect phase II or phase III or the MMC in the duodenum. Li+ inhibited pancreatic juice outflow as well as pancreatic enzyme and bicarbonate output (P < 0.05 for all pancreatic parameters).

Hydration properties and phosphorous speciation in native, gelatinized and enzymatically modified potato starch analyzed by solid-state MAS NMR.

Hydration of granular, gelatinized and molecularly modified states of potato starch in terms of molecular mobility were analyzed by (13)C and (31)P solid-state MAS NMR. Gelatinization (GEL) tremendously reduced the immobile fraction compared to native (NA) starch granules. This effect was enhanced by enzyme-assisted catalytic branching with branching enzyme (BE) or combined BE and ß-amylase (BB) catalyzed exo-hydrolysis. Carbons of the glycosidic α-1,6 linkages required high hydration rates before adopting uniform chemical shifts indicating solid-state disorder and poor water accessibility. Comparative analysis of wheat and waxy maize starches demonstrated that starches were similar upon gelatinization independent of botanical origin and that the torsion angles of the glycosidic linkages were averages of the crystalline A and B type structures. In starch suspension phosphorous in immobile regions was only observed in NA starch. Moreover phosphorous was observed in a minor pH-insensitive form and as major phosphate in hydrated GEL and BE starches.

Plasma and urine concentrations of bioactive dietary benzoxazinoids and their glucuronidated conjugates in rats fed a rye bread-based diet.

Thorough knowledge of the absorption and metabolism of dietary benzoxazinoids is needed to understand their health-promoting effects. In this study, the fates of these bioactive compounds were examined by LC-MS/MS in plasma, urine, and feces after ingesting a daily dose of 4780 ± 68 nmol benzoxazinoids from rye bread using Wistar rats as a model. HBOA-glc (2-ß-D-glucopyranosyloxy-1,4-benzoxazin-3-one) was the predominant benzoxazinoid in the plasma (74 ± 27 nmol/L), followed by DIBOA-glc (2-ß-D-glucopyranosyloxy-4-hydroxy-1,4-benzoxazin-3-one) and HBOA. The total level of benzoxazinoids in the urine was 1176 ± 66 nmol/d, which corresponds to approximately 25% of the total dietary intake. The urinary benzoxazinoid profile differed from that of plasma with HBOA-glc and DIBOA-glc (647 ± 31 and 466 ± 33 nmol/d, respectively) as the major urinary components. The glucuronide conjugates of HBOA and DIBOA were detected in both the plasma and urine. N-dehydroxylation was found to be a critical step in the absorption of hydroxamic acids. This unprecedented study will trigger future interest in the biological effects of benzoxazinoids in whole grain rye and wheat diets in humans and other animals.

Bioactive benzoxazinoids in rye bread are absorbed and metabolized in pigs.

Recently, bioactive benzoxazinoids were discovered in cereal grains and bakery products. In this study, we studied the uptake, distribution, and metabolism of these secondary metabolites using a pig model. Twelve benzoxazinoid compounds and their 4 transformation products were quantified in the pigs' diets and biofluids using high-performance liquid chromatography coupled to electrospray ionization triple quadrupole mass spectrometry. The 2-ß-D-glucopyranosyloxy-4-hydroxy-1,4-benzoxazin-3-one (DIBOA-glc) was the most dominant benzoxazinoid (232 nmol/g DM) seconded by the double-hexose derivative of DIBOA (provisionally characterized here as DIBOA-glc-hex) in the rye-based diet. DIBOA-glc (derived from the diet and intestinal deglycosylation of DIBOA-glc-hex) was apparently reduced to 2-ß-D-glucopyranosyloxy-1,4-benzoxazin-3-one (HBOA-glc), the most dominant benzoxazinoid in the blood (829 nmol/L). The benzoxazinoid compounds were excreted in the urine, with HBOA-glc (18 µmol/L) as a major metabolite. In this study, we determined for the first time the bioavailability of dietary benzoxazinoids that have high digestibility, distribution, and metabolism in mammals. These findings could be a milestone for the exploitation of healthful and pharmacological properties of benzoxazinoids.

Quantitative aspects of the metabolism of lignans in pigs fed fibre-enriched rye and wheat bread.

A diet rich in lignans has been suggested to be protective against a range of chronic diseases. The distribution and metabolic fate of lignans is, however, very poorly understood. We fed high-fibre wheat breads low in lignans (n 8) or high-fibre rye breads (n 9) rich in plant lignans to pigs for 58-67 d, and analysed the content of plant lignans and their metabolites in the diet, blood, bile, faeces, urine and selected tissues. Apparent faecal digestibility of dietary precursors was higher than of total (plant- and entero-) lignans due to conversion to enterolactone and enterodiol. The digestibility of lariciresinol and matairesinol was lower than that of the sum of plant lignans. This suggests that interconversion of plant lignans during digestion and enterohepatic circulation occur without complete conversion to enterolignans. The majority of lignans present in plasma and urine was in the form of enterolignans, but up to 23 % in the plasma, and 11 % in the urine of the rye-fed pigs were in the form of plant lignans. There was a very high concentration of lignans in bile from the rye-fed pigs with as much as 77 % in the form of plant lignans. Lignans were detected in the tissue of colon, liver, breast and brain at a much higher level with rye than with wheat, but only in the form of enterolactone. The importance and implications of systemic exposure to plant lignans remain to be elucidated.

Metabolic profiling of liver from hypercholesterolemic pigs fed rye or wheat fiber and from normal pigs. High-resolution magic angle spinning 1H NMR spectroscopic study.

This study presents the first application of a high-resolution magic angle spinning 1H NMR approach to elucidate the metabolic effects of a hypercholesterolemic condition and two high-fiber diets based on rye and wheat bread, respectively, in intact pig liver biopsy samples. Standard 1D and spin-echo 1H spectra were acquired on a total of 20 biopsy samples, and 2D total correlation spectroscopy experiments were carried out on selected samples for assignment of the observed resonances. Principal component analyses and partial least-squares regression discriminant analysis revealed differences in the hepatic lipid content and choline-containing compounds between normal and hypercholesterolemic pigs. In addition, the results demonstrated that the liver metabolite profile of hypercholesterolemic pigs fed a high-fiber rye bread differed from that of pigs fed high-fiber wheat bread with respect to both the lipoprotein fractions and the choline-containing compounds. These findings suggest that earlier reports on high-fiber rye diet-induced effects on plasma HDL/LDL content partially can be ascribed to effects on liver cholesterol metabolism and that the hepatic phospholipase pathways of phosphatidylcholine breakdown are affected by the high-fiber rye diet.

Protein-polyphenol interactions and in vivo digestibility of buckwheat groat proteins.

A rat model system, based on a diet with (Nb+) or without (Nb-) the inclusion of 0.7% of a 2:1 (w/w) mixture of Bacitracin and Neomycin sulfate (Nebacitin) was used to elucidate interactions of polyphenols with proteins in buckwheat. The treatment with the antibiotic Nebacitin suppresses the microflora in the large intestine and is used to obtain an estimate of the digestion occurring in the small intestine. Experimental buckwheat samples were obtained in order to contain different levels of polyphenol and were subjected to different hydrothermal treatments. It was found that a considerable interaction between polyphenols and proteins appeared during the hydrothermal treatment, and this interaction reduces the digestion of proteins through the small and large intestine. Microbial processes in the colon enhance the digestibility of protein, blocked by polyphenols in hydrothermally processed buckwheat.