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1
Liu, S. Y., and P. H. Selle. "Starch and protein digestive dynamics in low-protein diets supplemented with crystalline amino acids." Animal Production Science 57, no. 11 (2017): 2250. http://dx.doi.org/10.1071/an17296.
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Digestive dynamics of starch and protein is defined as digestion of starch and protein, absorption of glucose and amino acids from the gut lumen and their transition across enterocytes to enter the portal circulation. Digestive dynamics consider the extent, rate and site of nutrient digestion along the small intestine and the bilateral, post-enteral bioavailability of glucose and amino acids. The underlying premise is that glucose and amino acids should be made available in appropriately balanced quantities at the sites of protein synthesis for efficient protein deposition and growth performance. Previous studies have suggested that feed conversion efficiency may be enhanced by rapidly digestible protein and that crystalline amino acids could be considered sources of ‘rapid protein’. At present, crystalline lysine, methionine and threonine are routinely included in broiler diets; moreover, an increasing array of both essential and non-essential crystalline amino acids is becoming commercially available. Despite unrestricted feed access in commercial chicken-meat production systems during the period of illumination, it appears that the intermittent feed consumption patterns of broiler chickens still provide scope for asynchronies in digestion and absorption of nutrients, which affects broiler performance. A better understanding of the post-enteral, bilateral bioavailability of glucose and amino acids in low-protein diets containing high levels of synthetic amino acids is clearly desirable and this applies equally to the relationship between crystalline and protein-bound amino acids.
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Truong, Ha H., Sonia Y. Liu, and Peter H. Selle. "Starch utilisation in chicken-meat production: the foremost influential factors." Animal Production Science 56, no. 5 (2016): 797. http://dx.doi.org/10.1071/an15056.
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Starch is the chief dietary energy source for chicken-meat production, the majority of which is derived from the grain basis of diets for broiler chickens. The utilisation of starch from maize is of a high order in terms of ileal starch digestibility coefficients but this is not necessarily the case with wheat or sorghum. This may stem from the fact that maize essentially lacks the soluble non-starch polysaccharides in wheat and ‘non-tannin’ phenolic compounds found in sorghum. Numerous factors may influence starch digestibility with emphasis placed on starch–protein interactions as starch granules are located in the prolamin protein matrixes of grain endosperm. This close proximity facilitates any physical and chemical interactions and in this connection particular attention has been paid to kafirin, the dominant protein fraction in sorghum. Nevertheless, despite their apparent importance, the precise nature of starch–protein interactions has not been well defined. Exogenous phytases are routinely included in broiler diets primarily to liberate phytate-bound phosphorus; however, phytate may impede starch digestion and may retard glucose absorption. Additional feed additives, including non-starch polysaccharide-degrading enzymes, other exogenous enzymes and reducing agents may have the capacity to influence starch utilisation. Nevertheless, ileal and total tract starch digestibility coefficients are static parameters and overlook the digestive dynamics of starch, which is inappropriate given the possibility that slowly digestible starch enhances energy utilisation and feed conversion efficiency. However, if the slowly digestible starch concept is valid, the underlying mechanisms have not been fully elucidated. Consideration is given to the suggestion that slowly digestible starch ameliorates the catabolism of amino acids to provide energy to the gut mucosa by increasing the provision of glucose to posterior small intestinal segments. There is the prospect that whole grain feeding provides slowly digestible starch in addition to generating heavier relative gizzard weights. The digestive dynamics of starch and protein are inter-related and the digestion of starch and absorption of glucose should not be considered in isolation from protein digestion and amino acid absorption in the quest to improve the performance of broiler chickens. The foremost factor influencing starch utilisation in chicken-meat production may be the interaction between starch and protein digestive dynamics.
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Scheppach, Wolfgang, Hardi Luehrs, and Thomas Menzel. "Beneficial health effects of low-digestible carbohydrate consumption." British Journal of Nutrition 85, S1 (March 2001): S23—S30. http://dx.doi.org/10.1079/bjn2000259.
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Low-digestible carbohydrates represent a class of enzyme-resistant saccharides that have specific effects on the human gastrointestinal tract. In the small bowel, they affect nutrient digestion and absorption, glucose and lipid metabolism and protect against known risk factors of cardiovascular disease. In the colon they are mainly degraded by anaerobic bacteria in a process called fermentation. As a consequence, faecal nitrogen excretion is enhanced, which is used clinically to prevent or treat hepatic encephalopathy. Low-digestible carbohydrates are trophic to the epithelia of the ileum and colon, which helps to avoid bacterial translocation. Short-chain fatty acids are important fermentation products and are evaluated as new therapeutics in acute colitis. They are considered in the primary prevention of colorectal cancer. The bifidogenic effect of fructo-oligosaccharides merits further attention. Unfermented carbohydrates increase faecal bulk and play a role in the treatment of chronic functional constipation, symptomatic diverticulosis and, possibly, the irritable bowel syndrome. In conclusion, low-digestible carbohydrates may play a role in the maintenance of human digestive health. However, the strength of evidence differs between disease entities.
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Rothman, Stephen, Charles Liebow, and Lois Isenman. "Conservation of Digestive Enzymes." Physiological Reviews 82, no. 1 (January 1, 2002): 1–18. http://dx.doi.org/10.1152/physrev.00022.2001.
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The traditional understanding is that an entirely new complement of digestive enzymes is secreted by the pancreas into the small intestines with each meal. This is thought to be necessary because, like food itself, these enzymes are degraded during digestion. In this review we discuss experiments that bring this point of view into question. They suggest that digestive enzymes can be absorbed into blood, reaccumulated by the pancreas, and reutilized, instead of being reduced to their constituent amino acids in the intestines. This is called an enteropancreatic circulation of digestive enzymes.
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Lebedev, Sviatoslav Valerievich, Elmira Zakievna Gubaidulina, Elena Vladimirovna Sheida, and Victoria Vladimirovna Grechkina. "Exchange (uptake and synthesis) of amino acids in the digestive tract of cattle when used in diet different ingredient composition of the feed." Agrarian Scientific Journal, no. 4 (April 29, 2019): 54–57. http://dx.doi.org/10.28983/asj.y2019i4pp54-57.
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Materials characterizing the role of digestive tract in amino acid metabolism are obtained based on studies assessing the effect of diets with various sources of protein (fish meal, sunflower meal). A number of metabolic regularities (synthesis and assimilation) of amino acids in the digestive tract of an animal follow from the data obtained by us. Сhyme inflowing from the stomach into intestine, contains 1.5-2 times more amino acids, compared with the number of them in the diet. Consequently, the same amount of basic amino acids can be synthesized in the process of digestion of ruminants. Most of amino acids from chyme is digested in the intestine. The actual amount of amino acids digested and included in the metabolism of animal was significantly higher than their content in the eaten dietand reached 108.1 - 148.9% of that received with the diet.
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Longstaff, Margaret, and J. M. McNAB. "The inhibitory effects of hull polysaccharides and tannins of field beans (Vicia faba L.) on the digestion of amino acids, starch and lipid and on digestive enzyme activities in young chicks." British Journal of Nutrition 65, no. 2 (March 1991): 199–216. http://dx.doi.org/10.1079/bjn19910081.
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The effects of polysaccharides and tannins present in the hulls of field beans (Vicia faba L.) on the digestion of amino acids, starch and lipid were studied in poultry. A control diet without hulls and the same diet substituted with 400 g hulls/kg diet from three different varieties of beans were fed to 3-week-old chicks for 4 d. Digestibility coefficients for amino acids, starch and lipid were calculated from measurements made of these nutrients in the diets and the freeze-dried excreta with the aid of titanium dioxide as a marker. Activities of trypsin (EC 3.4.21.4), α-amylase (EC 3.2.1.1), and lipase (EC 3.1.1.3) in digesta removed from the upper jejunum, sucrase (EC 3.2.1.48) in the gut mucosa from the upper jejunum, and α-amylase and lipase in the pancreas were measured. The hulls were analysed for their polysaccharide and tannin contents. Results showed that the hulls were mostly carbohydrate in composition, with cellulose the predominant polysaccharide. Tannins present in the hulls of two coloured-flowering varieties (Brunette and Minica) were of the condensed type. The diet with tannin-free hulls (white-flowering variety Medes) lowered slightly the digestion of amino acids, starch and lipid compared with the control diet. This effect was believed to be due to inhibition of digestive enzymes, possibly through their adsorption onto the hulls. Diets with tannin-rich hulls (varieties Brunette and Minica) caused a large reduction in the digestion of amino acids, starch and lipid compared with the control diet mainly due to inactivation of digestive enzymes by the formation of tannin–enzyme complexes in the digestive tract. Enzyme activities could be partially restored by the addition of polyvinylpyrrolidone to the digesta. Tannins inactivated trypsin the most, α-amylase to a lesser extent and lipase the least and as a consequence lowered the digestion of amino acids the most, starch to a lesser extent and lipid the least. Tannins did not induce an increased pancreatic production of digestive enzymes, nor did they affect activity of jejunum mucosal sucrase. Condensed tannins from Brunette and Minica hulls were partially extractable in methanol alone, but required acidic methanol for fuller extraction. The vanillin: anthocyanidin ratio suggested that tannins were polymerized to the same degree in the Brunette and Minica varieties, both in the methanol and acidic methanol extracts. Hulls from the variety Minica contained a greater amount of methanol-extractable tannins, the quantity of remaining tannins that required acidic methanol for extraction being the same for both varieties.
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Stevens, J., I. Chivers, D. Symons, and K. Dixon. "Acid-digestion improves native grass seed handling and germination." Seed Science and Technology 43, no. 2 (August 1, 2015): 313–17. http://dx.doi.org/10.15258/sst.2015.43.2.19.
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Taghizadeh, A. T., B. Najafnejad, and R. Noori. "Evaluation of nutritive value of tropical clover (Trifolium resupinatum) using gas production technique." Proceedings of the British Society of Animal Science 2007 (April 2007): 217. http://dx.doi.org/10.1017/s1752756200021207.
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Gas measurement provides a useful data on digestion kinetics of both soluble and insoluble fractions of feedstuffs (Getachew et al., 1998). Carbohydrate portion of each feed is divided into three digestible fraction: the A fraction, containing sugars, short oligosaccharides, and organic acids; the B1 fraction, containing starch and pectin; and B2, the digestible fibre fraction (Doane et al. 1998). The objective of this study was to measure of fermentation parameters of tropical clover (Trifolium resupinatum) (pre bud, full bud, pre bloom and full bloom) using gas production technique.
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Larder, Christina E., Michèle M. Iskandar, and Stan Kubow. "Dynamic Multi-Stage Gastrointestinal Digestion Model Assessment of Microbial Fermentation Products of Collagen Hydrolysates." Proceedings 61, no. 1 (October 30, 2020): 12. http://dx.doi.org/10.3390/iecn2020-06998.
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Proteins, peptides and amino acids (AAs) that bypass upper gastrointestinal (GI) digestion can be fermented in the colonic regions. This could lead to microbial production of health promoting short-chain fatty acids (SCFAs). Nitrogenous compounds can also be fermented to generate potentially harmful branched chain fatty acids (BCFAs). As collagen hydrolysate (CH) supplements contain a high peptide content, we evaluated whether peptides that undergo intestinal CH digestion and microbial fermentation can generate SCFAs and BCFAs. Two bovine-sourced CH formulations (CH-GL and CH-OPT) underwent digestive processes and microbial fermentation for 24 h in a dynamic GI digestion model containing human fecal matter. After 24 h, CH-OPT showed a significant (p < 0.05) increase in SCFAs (propionic, butyric and valeric acids) in the ascending colonic vessel with no changes observed with CH-GL. Only CH-OPT showed a significant (p < 0.05) increase in BCFAs, also noted in the ascending colon. No significant (p < 0.05) changes to SCFAs and BCFAs were observed in the transverse and descending colons for both CHs. These findings demonstrate that CHs can induce microbial production of SCFAs and BCFAs although this appears to depend on the CH tested. More studies are needed to determine the physiological significance of these microbial metabolites from intake of CH supplements.
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Vong, M. H., and M. L. Stewart. "In vitro bacterial fermentation of tropical fruit fibres." Beneficial Microbes 4, no. 3 (September 1, 2013): 291–95. http://dx.doi.org/10.3920/bm2013.0006.
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Tropical fruits such as mango, papaya, pineapple and banana are rich sources of dietary fibre. However, few studies have examined the potential physiological effects of fibre from these tropical fruits. The aim of this study was to characterise the fermentability of dietary fibre found in banana, papaya, pineapple and mango as an estimate of the physiological effects of consuming these fruits. Freeze-dried fruit was subjected to in vitro digestion to remove digestible carbohydrates. Digestion residues were freeze-dried prior to fermentation. In vitro fermentation was carried for 24 h under anaerobic conditions to simulate conditions in the large intestine. Gas volume, pH and short chain fatty acids (SCFAs) concentration were measured at 0, 4, 8, 12, and 24 h. SCFAs were analysed by gas chromatography. There was no gas production from 0 to 8 h time points for all samples. Mango fibre resulted in more gas at 12 and 24 h than pineapple, papaya and banana fibres. The slurry pH was significantly lower for mango fibre at 12 and 24 h compared to other samples. Mango fibre resulted in significantly more propionate at 8 h compared to papaya and pineapple fibres. Butyrate concentrations were only significantly different at 4 h. At 24 h total and individual SCFA production did not differ among samples. All fruit fibres were fermentable, with mango fibre being the most rapidly fermented. Additional work is necessary to confirm a benefit on digestive health.
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Heras, Joseph, Mahul Chakraborty, J. J. Emerson, and Donovan P. German. "Genomic and biochemical evidence of dietary adaptation in a marine herbivorous fish." Proceedings of the Royal Society B: Biological Sciences 287, no. 1921 (February 19, 2020): 20192327. http://dx.doi.org/10.1098/rspb.2019.2327.
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Adopting a new diet is a significant evolutionary change, and can profoundly affect an animal's physiology, biochemistry, ecology and genome. To study this evolutionary transition, we investigated the physiology and genomics of digestion of a derived herbivorous fish, Cebidichthys violaceus . We sequenced and assembled its genome (N50 = 6.7 Mb) and digestive transcriptome, and revealed the molecular changes related to digestive enzymes (carbohydrases, proteases and lipases), finding abundant evidence of molecular adaptation. Specifically, two gene families experienced expansion in copy number and adaptive amino acid substitutions: amylase and carboxyl ester lipase ( cel ), which are involved in the digestion of carbohydrates and lipids, respectively. Both show elevated levels of gene expression and increased enzyme activity. Because carbohydrates are abundant in the prickleback's diet and lipids are rare, these findings suggest that such dietary specialization involves both exploiting abundant resources and scavenging rare ones, especially essential nutrients, like essential fatty acids.
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Kara, K. "Estimated ruminal digestion values and digestion end-products of concentrated mix feed after in vitro treatment with propionic acid." Veterinární Medicína 63, No. 12 (December 3, 2018): 537–45. http://dx.doi.org/10.17221/100/2017-vetmed.
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This study was aimed at determining the effects of propionic acid supplementation at doses of 0 (control group, PA0), 12, 24, 48 and 96 mM (PA12, PA24, PA48, and PA96) to concentrated mix feed on in vitro cumulative total gas production, methane emission, gas kinetics (potential gas production, (a + b)<sub>gas</sub> and gas production rate, c<sub>gas</sub>), estimated digestibility, estimated energy value and the end-products and variables of in vitro digestion (total bacteria count, the number of ciliate protozoa, volatile fatty acids, pH value and ammonia-N). Digestion treatments were carried out in an anaerobic in vitro fermenter for up to 96 h. The in vitro cumulative total gas production, (a + b)<sub>gas</sub>, estimated metabolic energy, estimated net energy lactation and estimated organic matter digestibility and ammonia-N concentration were decreased by propionic acid up to 96 mM (P < 0.05). In the in vitro fermenter fluid, total bacteria count, the total numbers of ciliate protozoa and the individual numbers of some ciliate protozoa (Entodiniinae, Isotricha spp. and Diplodiniinae) (P < 0.01) decreased linearly with increasing concentrations of dietary propionic acid. The total molar concentrations of volatile fatty acids decreased in response to propionic acid supplementation (P < 0.001). Dietary propionic acid elicited linear increases in the molar concentrations of propionic acid (P < 0.001) and butyric acid (P < 0.01) as proportions of total volatile fatty acids of the in vitro fermenter fluid. In contrast, molar proportions of acetic acid, the c<sub>gas</sub>, pH values and the numbers of Dasytricha sp. were not affected by dietary propionic acid supplementation (P > 0.05). The addition of 12–96 mM propionic acid to concentrated mix feed decreased methane emission from the rumen and negatively affected microbiota count, feed digestibility, proteolysis, and molar volatile fatty acid values in the rumen environment.
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Boila, R. J., and J. R. Ingalls. "Prediction of rumen undegradable amino acids that are digested post-ruminally." Canadian Journal of Animal Science 75, no. 4 (December 1, 1995): 583–92. http://dx.doi.org/10.4141/cjas95-087.
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Data for in situ disappearance of DM, nitrogen (N) and amino acids in canola meal (CM) were used to determine degradation parameters required to estimate effective degradabilities of these nutrients. The non-digestible fraction of DM, N and amino acids in CM was determined using mobile bags after ruminal incubation for 12 h. Linear relationships were determined between dependent variables, ruminal disappearance (DIS) or effective degradability (EDEG) of amino acids and independent variables, DIS or EDEG of DM or N. Use of EDEG vs. EDEG relationships, (r2 greater than 0.87 for amino acids other than methionine and cystine) with the effective degradability of DM, as the independent variable, was the most effective approach over a wide range of rumen outflow rates (k) for prediction of amino acids that escaped ruminal degradation. Use of DIS vs. DIS (an estimate for EDEG of DM served as a value for the independent variable), or EDEG vs. DIS (disappearance of DM at incubation time equal to k−1 was the independent variable) relationships was not as effective. Post-ruminal digestion was predicted as the difference between ruminal escape (based on estimates of effective degradability) and that nutrient present in the residual DM in mobile bags recovered at the terminal ileum. Use of linear EDEG vs. EDEG equations, in association with a mobile bag technique, allowed for prediction of both ruminal escape and post-ruminal digestion of N and amino acids in CM over a range of outflow rates for feed particles from the rumen. Key words: Amino acids, canola meal, ruminal escape, post-ruminal digestion
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Livesey, Geoffrey. "The absorption of stearic acid from triacylglycerols: an inquiry and analysis." Nutrition Research Reviews 13, no. 2 (December 2000): 185–214. http://dx.doi.org/10.1079/095442200108729061.
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AbstractAlthough stearic acid is a saturated fatty acid, its influence on plasma cholesterol acid other health variables is neutral; possibly owing in part to poor absorption. Reduced absorption of stearic acid from particular triacylglycerols, cocoa butter and novel fats formulated with short- and long-chain acid triacylglycerol molecules (Salatrims) has been attributed to high intakes. However, the circumstances and causes of poor stearic acid digestion from triacylglycerols are unclear; published data were therefore collected and analysed, with emphasis on human studies. Of twenty-eight studies conducted in adults, most are in men (>90%). The assertion that reduced absorption is due to a high intake of stearoyl groups is not supported: dietary intakes of stearoyl of 0·05–0·65 g stearic acid equivalent/kg body weight (cf typical intake of 0·2 g stearic acid equivalent/kg body weight in the Western diet) indicate that the ‘true’ digestibility of stearoyl is 0·98 (SE 0·01) g/g, with apparent digestibility less than this value at low intakes owing to endogenous stearic acid excretion and to inter-publication variation of unidentified cause. The neutral health impact of stearic acid must be due to factors other than availability. Exceptions include cocoa butter, Salatrims and tristearin, for which digestibility is an additional factor. The efficiency with which human subjects digest stearoyl from cocoa butter still remains uncertain, while the digestion of total long-chain fat from this source is 0·89–0·95 g/g, high in comparison with 0·33 g/g for Salatrim 23CA and 0·15 g/g for tristearin in their prepared states. Salatrims contain the highest proportion of long-chain fatty acids that are stearic acid-rich other than tristearin, which is the main component of fully-hydrogenated soyabean and rapeseed oil. Analysis shows that apparent digestibility of stearic acid is associated with stearoyl density within the triacylglycerol molecule and that, in Salatrims, the occurrence of short-chain fatty acids in place of long-chain fatty acids increases this density. Soap formation appears not to be a major factor in the reduced digestion of stearic acid from tristearin under regular dietary circumstances, but both microcrystallinity and reduced digestibility of tri-, di- and monostearoylglycerols appears to be important. Solubilisation of high-melting-point tristearin in low-melting-point oils improves the digestibility of its stearic acid, particularly when emulsified or liquidized at above melting point. However, without such artificial aids, the digestive tracts of the rat, dog and man have a low capacity for emulsifying and digesting stearic acid from tristearin. Reduced digestibility of stearic acid from Salatrim 23CA also appears to be attributable to reduced digestibility of di- and monostearoylglycerols and is particularly due to remnants with the 1- or 3-stearoylglycerol intact after initial hydrolytic cleavage. Short-chain organic acid in Salatrim 23CA, which is readily hydrolysed, leaves such remnants. Unlike tristearin, Salatrim 23CA melts at body temperature and mixing it with low-melting-point oils is not expected to cause further disruption of microcrystalline structures to aid digestibility of its stearoyl groups. The low digestibility of stearoyl in Salatrim 23CA, together with the occurrence of short-chain organic acids in this product, account for its relatively low nutritional energy value (about 20 kJ (5 kcal)/g) compared with traditional fats (37 kJ (9 kcal)/g) and low fat value (<20:37 kJ/kJ; <5:9 kcal/kcal) relative to traditional fats. In part these differences are because of minor effects of Salatrim 23CA on the excretion of other fat and protein, due to the bulking properties of this poorly-digestible fat.
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Zhou, Yuyin, and Chi Chen. "Formation of Strecker Aldehydes in Casein Digestion." Current Developments in Nutrition 4, Supplement_2 (May 29, 2020): 500. http://dx.doi.org/10.1093/cdn/nzaa045_133.
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Abstract Objectives This study investigated the occurrence of Strecker degradation during in vitro digestion. Methods Casein was first incubated with artificial gastric fluid containing porcine pepsin and HCl (pH = 2) for 60 min. After adjusting pH to 7 with sodium hydroxide, casein gastric digesta was then incubated with artificial intestinal fluid containing porcine pancreatin (pH = 7) for 120 min. Digesta samples were collected at 0, 10, 20, 30, and 60 min of gastric digestion, and then 10, 20, 30, 60, and 120 min of intestinal digestion. Free amino acids and aldehydes in digesta samples were analyzed by the liquid chromatography-mass spectrometry analysis. Results Multiple aldehydes were detected in gastric digestion samples, and their concentrations were further increased by intestinal digestion. Among them, isovaleraldehyde, isobutyraldehyde, phenylacetaldehyde, and acetaldehyde, are the Strecker degradation products of leucine, valine, phenylalanine, and alanine, respectively. Without digestive enzymes, casein incubation did not produce Strecker aldehydes. Conclusions In vitro digestion of proteins can produce Strecker aldehydes. Funding Sources This research was partially supported by the Agricultural Experiment Station project MIN-18-125 (C. C.) from the United States Department of Agriculture (USDA).
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Shrestha, Ashok Kumar. "Scientific Background of Dairy Protein Digestibility: A Review." Journal of Food Science and Technology Nepal 7 (June 7, 2014): 1–8. http://dx.doi.org/10.3126/jfstn.v7i0.10560.
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Recent advances have shown that differences in compositional, structural and physical properties of caseins and whey proteins affect their digestion and absorption behavior, hormonal response, satiety effect and other physiological effects. For example, the ingestion of whey protein cause fast, high and transient increase of amino acids ‘fast protein’, whereas casein induce slower, lower and prolonged increase of ‘slow protein’ in the gut. Knowledge of, and control over, the rate and nature of digestive breakdown of dairy proteins provides a potential basis for product/process innovation through identifying ingredients and formulations that provide desired nutrient delivery profiles. With this background, the aim of our current review paper is to understand the digestion behavior of various protein-rich milk powders and their potential use in formulation of dairy foods for controlled release of amino acids and energy. Currently available in vitro protein digestibility methods to measure or predict the dairy protein digestibility were also investigated. The author has also presented the preliminary results of ongoing study on in vitro digestion of various commercial proteins powders.DOI: http://dx.doi.org/10.3126/jfstn.v7i0.10560 J. Food Sci. Technol. Nepal, Vol. 7 (1-8), 2012
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Kouns, WC, PJ Newman, KJ Puckett, AA Miller, CD Wall, CF Fox, JM Seyer, and LK Jennings. "Further characterization of the loop structure of platelet glycoprotein IIIa: partial mapping of functionally significant glycoprotein IIIa epitopes." Blood 78, no. 12 (December 15, 1991): 3215–23. http://dx.doi.org/10.1182/blood.v78.12.3215.3215.
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Abstract Glycoprotein (GP) IIb-IIIa serves as the platelet fibrinogen receptor. Studies of the tertiary structure of GPIIIa have shown that the protein has a large loop structure of at least 325 amino acids in length. To further characterize this loop structure, intact platelets were digested with alpha-chymotrypsin. Digestion products were examined using the anti-GPIIIa monoclonal antibodies (MoAbs) AP3, D3GP3, and C5GP3, as well as the human alloantibody, anti-PLA1. AP3 recognized GPIIIa digestion products of 109, 95, and 68 Kd. D3GP3 and C5GP3 recognized an additional band of 51 Kd. Time course digestions demonstrated that the 51-Kd fragment was generated by proteolysis of the 68-Kd peptide. Sequence analysis of the reduced 51-Kd peptide showed that this fragment began at amino acid 422. The nonreduced 51-Kd peptide was reactive with antibodies directed against the first 13 amino acids of GPIIIa, demonstrating the presence of a covalently attached N-terminal peptide. These data suggest that: (1) the minimum length of the loop structure is at least 384 amino acids; (2) the AP3 epitope is formed at least in part by a determinant contained within residues 348 to 421; and (3) the D3GP3 and C5GP3 epitopes are contained within amino acids 422 to 692 of GPIIIa, a region that may be flexible and involved in conformational changes that occur after ligand binding.
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Kouns, WC, PJ Newman, KJ Puckett, AA Miller, CD Wall, CF Fox, JM Seyer, and LK Jennings. "Further characterization of the loop structure of platelet glycoprotein IIIa: partial mapping of functionally significant glycoprotein IIIa epitopes." Blood 78, no. 12 (December 15, 1991): 3215–23. http://dx.doi.org/10.1182/blood.v78.12.3215.bloodjournal78123215.
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Glycoprotein (GP) IIb-IIIa serves as the platelet fibrinogen receptor. Studies of the tertiary structure of GPIIIa have shown that the protein has a large loop structure of at least 325 amino acids in length. To further characterize this loop structure, intact platelets were digested with alpha-chymotrypsin. Digestion products were examined using the anti-GPIIIa monoclonal antibodies (MoAbs) AP3, D3GP3, and C5GP3, as well as the human alloantibody, anti-PLA1. AP3 recognized GPIIIa digestion products of 109, 95, and 68 Kd. D3GP3 and C5GP3 recognized an additional band of 51 Kd. Time course digestions demonstrated that the 51-Kd fragment was generated by proteolysis of the 68-Kd peptide. Sequence analysis of the reduced 51-Kd peptide showed that this fragment began at amino acid 422. The nonreduced 51-Kd peptide was reactive with antibodies directed against the first 13 amino acids of GPIIIa, demonstrating the presence of a covalently attached N-terminal peptide. These data suggest that: (1) the minimum length of the loop structure is at least 384 amino acids; (2) the AP3 epitope is formed at least in part by a determinant contained within residues 348 to 421; and (3) the D3GP3 and C5GP3 epitopes are contained within amino acids 422 to 692 of GPIIIa, a region that may be flexible and involved in conformational changes that occur after ligand binding.
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Liu, Hui, Qian Li, and Chunbao Li. "Changes in the Structure and Digestibility of Myoglobin Treated With Sodium Chloride." Current Developments in Nutrition 5, Supplement_2 (June 2021): 661. http://dx.doi.org/10.1093/cdn/nzab045_043.
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Abstract Objectives Changes in the structure of myoglobin affect its digestibility and myoglobin can't be completely broken down of pepsin. The exact mechanism of this breakdown is not clear. It can be inferred that myoglobin is broken down into polypeptides whose structures do not fully conform to the spatial characteristics of digestive enzymes. Myoglobin dissolution in different salt concentrations and changes in secondary structure were examined by spectroscopic examination. The molecular dynamics simulation was used to study the stability of these structural changes and their combination with digestive enzymes. Finally, since the products of digestion are determined by the catalytic centers of digestive enzymes, the substrate channel is extended to study the relationship between digestive enzymes and substrates. Methods In this study, different concentrations of sodium chloride were added for a certain period of time in advance to detect changes in the secondary structure of myoglobin, leading to changes in digestibility. Myoglobin and digestive enzymes were docked for molecular dynamics simulations to analyze the energy and structural changes in the interactions between substrates and proteins. Results Salt-treated protein can affect the secondary structure changes of protein. High concentration of salt-treated protein will lead to protein aggregation and denaturation, affecting digestibility. Low concentration of salt-treated protein may lead to exposure of sleeping areas, reducing digestibility. In terms of structure, salt ions lead to changes in the bonding of amino acids in the protein and affect the substrate's entry into the enzyme activity center. Conclusions Treatment of myoglobin with different salt concentrations did not change its tertiary structure but low salt concentrations lowered its digestibility, probably due to salt ions altering a number of key amino acid bonds, making the structure more stable and less susceptible to digest. Some low molecular weight peptides remain after pepsin digestion. This may be due to the distance between the key amino acids in the active catalytic center of pepsin being insufficient to form stable conformations with small peptides. Funding Sources This work was supported by the grants from NSFC (32072211).
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Vertiprakhov, V. G., and A. A. Grozina. "EXOCRINE PANCREATIC FUNCTION IN CHICKENS AS A RESULT OF ADDING FEED ACIDIFIERS IN THEIR DIET." Siberian Herald of Agricultural Science 48, no. 6 (January 24, 2019): 63–69. http://dx.doi.org/10.26898/0370-8799-2018-6-9.
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Feed acidifiers are used in animal diets for the prevention of proliferation of intestinal pathogenic microorganisms and resulting gastrointestinal digestive disorders. These additives, containing organic acids, have also been found to improve productivity and feed efficiency in poultry. There is information about correlation between digestive enzymes’ activity and intestinal microbiota of meattype chickens. However, the exact mechanism of the beneficial impact of organic acids on the digestion system still remains understudied. The paper presents the results of experiments conducted on Hisex White chicken with chronic fistulae of the main pancreatic duct, fed on a diet supplemented with an acidifier containing 2-furoic acid. No significant effect of this acidifier was found on the digestive pancreatic function. The dynamics analysis showed that the chickens’ secretion rate of pancreatic juice after postprandial 30 minutes dropped by over two times in the testing period when using acidifiers in their diet. After 150 minutes this rate was lower by 27.3% compared to the control group, which corresponds the neurochemical phase of secretory regulation. Analysis of enzyme dynamics (amylase, lipase and protease) showed a slight increase in the activity of proteases in pancreatic juice (by 1.2- 12.4%), compared to the control group, in the phase of complex-reflex regulation of pancreatic secretory activity related to the recognition of the taste qualities of the feed. Feed conversion ratio in the test group of broiler chickens increased by 1.52% when using acidifiers in the diet. The result of the study showed that the use of acidifiers has a beneficial effect on chickens’ gastrointestinal digestion as well as an inhibitive action on intestinal pathogens.
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Macfarlane, George T., and Sandra Macfarlane. "Bacteria, Colonic Fermentation, and Gastrointestinal Health." Journal of AOAC INTERNATIONAL 95, no. 1 (January 1, 2012): 50–60. http://dx.doi.org/10.5740/jaoacint.sge_macfarlane.
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Abstract The colonic microbiota plays an important role in human digestive physiology and makes a significant contribution to homeostasis in the large bowel. The microbiome probably comprises thousands of different bacterial species. The principal metabolic activities of colonic microorganisms are associated with carbohydrate and protein digestion. Nutrients of dietary and host origin support the growth of intestinal organisms. Short-chain fatty acids (SCFAs), predominantly acetate, propionate, and butyrate, are the principal metabolites generated during the catabolism of carbohydrates and proteins. In contrast, protein digestion yields a greater diversity of end products, including SCFAs, amines, phenols, indoles, thiols, CO2, H2, and H2S, many of which have toxic properties. The majority of SCFAs are absorbed from the gut and metabolized in various body tissues, making a relatively small but significant contribution to the body's daily energy requirements. Carbohydrate fermentation is, for the most part, a beneficial process in the large gut, because the growth of saccharolytic bacteria stimulates their requirements for toxic products associated with putrefaction, for incorporation into cellular proteins, thereby protecting the host. However, as digestive materials move along the gut, carbohydrates become depleted, which may be linked to the increased prevalence of colonic disease in the distal bowel.
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Galloway Sr., D. L., A. L. Goetsch, A. Patil, L. A. Forster Jr., and Z. B. Johnson. "Feed intake and digestion by Holstein steer calves consuming low-quality grass supplemented with lasalocid or monensin." Canadian Journal of Animal Science 73, no. 4 (December 1, 1993): 869–79. http://dx.doi.org/10.4141/cjas93-089.
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Holstein steer calves were fed late vegetative bermudagrass or mature bromegrass free choice in three Latin-square experiments to determine influences of ionophore type and level, supplementation with cereal grain, and source of low-quality grass on feed intake and digestion. In exp. 1, neither lasalocid nor monensin given at 0.5 mg kg−1 of body weight (BW) affected intake or digestion with either forage source. In exp. 2, steers received lasalocid or monensin at 0, 0.5 or 1.0 mg kg−1 of BW. Neutral detergent fiber digestibility was not depressed by ionophores and was slightly greater (P < 0.05) for ionophores at 1.0 than at 0.5 mg kg−1 of BW. Digestible organic matter intake with bermudagrass was slightly greater with ionophores at 1.0 than at 0.5 mg kg−1 of BW, but an opposite difference between ionophore levels occurred with bromegrass (ionophore level × forage source interaction; P < 0.07). The concentration of total volatile fatty acids in rumen fluid at 4 h post-feeding was lower (P < 0.05) for monensin than for lasalocid; the acetate/propionate ratio was decreased by ionophores and was lower for monensin than for lasalocid (P < 0.05). In exp. 3, lasalocid at 0.5 mg kg−1 of BW did not affect intake or digestion or interact with supplemental corn (7.2 g kg−1 of BW) or forage source. In conclusion, level of ionophore maximizing digestible organic matter intake may vary with characteristics of source of low-quality forage. However, when present, effects of ionophores on digestibility or energy intake may be relatively small. Key words: Cattle, ionophore, forage, intake, digestion
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Wong, Y. H., Ch P. Tan, K. Long, and K. L. Nyam. "In vitro simulated digestion on the biostability of Hibiscus cannabinus L. seed extract." Czech Journal of Food Sciences 32, No. 2 (April 22, 2014): 177–81. http://dx.doi.org/10.17221/222/2013-cjfs.
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We investigate the biostability of phenolic acids from a kenaf (Hibiscus cannabinus L.) seed extract using an in vitro model simulating the physicochemical (pH, temperature and bile salts) and biological (gastric and pancreatic enzymes) gastrointestinal conditions. Some of the antioxidants in the kenaf seed extract were not relatively stable in the intestinal phase of the gastrointestinal tracts. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity and total phenolic content (TPC) assays displayed similar trends as the biostability of phenolic acids, which decreased during the digestion process. The overall percentage loss of selected phenolic acids was 8.4–49.4% in the intestinal phase. For the overall in vitro digestion system, significant correlations between phenolic acids, total phenolic content (TPC) and antioxidant activity (DPPH) were obtained in all digestion phases with the exception of the gastric phase.
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KELLY, J. M., R. J. CHRISTOPHERSON, and R. J. EARLY. "APPARENT DIGESTIBILTY OF AMINO ACIDS AND OTHER NITROGENOUS COMPOUNDS IN THE SMALL INTESTINE OF WETHERS EXPOSED TO A COLD ENVIRONMENT." Canadian Journal of Animal Science 69, no. 4 (December 1, 1989): 921–29. http://dx.doi.org/10.4141/cjas89-106.
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Digestion studies were conducted with eight shorn yearling Suffolk wethers to assess the effects of environmental temperature on intestinal digestion of nonammonia nitrogen (NAN) and amino acid N. The wethers were cannulated in the rumen, abomasum, and terminal ileum and subsequently chronically exposed to either cold (0–2 °C) or warm (21–25 °C) temperatures in a crossover experiment. Sheep were fed a diet of chopped bromegrass hay and digesta flows through the abomasum and terminal ileum were estimated by reference to 103ruthenium-phenanthroline and 51chromium-EDTA. Microbial N contributions to total N in abomasal digesta were estimated with 35sulphur. During cold exposure, mean rumen ammonia concentration was reduced (P < 0.01) by 20%, while net appearance of NAN across the forestomach increased from 2.7 to 7.3 g d−1 (P < 0.05) resulting in a 14% increase in flow of NAN from the abomasum. The latter was associated with a 63% increase in flow of undegraded feed N (P < 0.05) and a nonsignificant decrease in microbial NAN. There was a small increase in flow of most amino acids from the abomasum due to cold exposure, with significant effects for lysine, histidine and tyrosine (P < 0.05). Disappearances of lysine (P < 0.05) and tyrosine (P < 0.005) in the small intestine were increased by the cold environment. Cold exposure increased small intestinal digestion of amino acid N relative to digestible organic matter intake (P < 0.08) suggesting maintenance of amino acid supply relative to that of energy in a cold environment. Key words: Amino acids, digestion, small intestine, sheep, cold environment
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Hogan, JP, PM Kennedy, CS McSweeney, and AC Schlink. "Quantitative studies of the digestion of tropical and temperate forages by sheep." Australian Journal of Experimental Agriculture 29, no. 3 (1989): 333. http://dx.doi.org/10.1071/ea9890333.
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Studies of the digestion by sheep of tropical grasses Digitaria decumbens and Setaria sphacelata and the temperate grass Lolium perenne grown at 27.5�S. latitude plus a clover Trifolium resupinatum grown in southern Australia are reported. There were no major differences between forages in the site of organic matter digestion, in protein flow from the stomach, in concentrations or proportions of rumen volatile fatty acids and in chewing behaviour between these data and the values generally reported for temperate forages of similar chemical composition and digestibility. However rumen ammonia concentrations for D. decumbens and S. sphacelata (53 and 34 mg N/L) were appreciably lower than predicted values of 108 and 131 mg N/L. Apart from this variable it appears that relationships developed with temperate feeds provide adequate prediction of digestive characteristics of tropical feeds.
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David, Aditi, Tanvi Govil, Abhilash Tripathi, Julie McGeary, Kylie Farrar, and Rajesh Sani. "Thermophilic Anaerobic Digestion: Enhanced and Sustainable Methane Production from Co-Digestion of Food and Lignocellulosic Wastes." Energies 11, no. 8 (August 8, 2018): 2058. http://dx.doi.org/10.3390/en11082058.
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This article aims to study the codigestion of food waste (FW) and three different lignocellulosic wastes (LW) (Corn stover (CS), Prairie cordgrass (PCG), and Unbleached paper (UBP)) for thermophilic anaerobic digestion to overcome the limitations of digesting food waste alone (volatile fatty acids accumulation and low C:N ratio). Using an enriched thermophilic methanogenic consortium, all the food and lignocellulosic waste mixtures showed positive synergistic effects of codigestion. After 30 days of incubation at 60 °C (100 rpm), the highest methane yield of 305.45 L·kg−1 volatile solids (VS) was achieved with a combination of FW-PCG-CS followed by 279.31 L·kg−1 VS with a mixture of FW-PCG. The corresponding volatile solids reduction for these two co-digestion mixtures was 68% and 58%, respectively. This study demonstrated a reduced hydraulic retention time for methane production using FW and LW.
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Ghosh, S., D. Jerger, M. P. Henry, and A. Sajjad. "Rapid-rate thermophilic, single-stage and two-phase methane fermentation of synfuel-industry wastewaters." Water Science and Technology 43, no. 1 (January 1, 2001): 35–42. http://dx.doi.org/10.2166/wst.2001.0009.
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Thermophilic (55°C) anaerobic digestion of wastewaters from wet carbonization of Minnesota peat was conducted in a simple, upflow digester at an HRT of one day to provide a methane yield of 0.20 SCM/kg VS added and a BOD5 reduction of 85%. Carbonization of Maine peat conducted for a much longer duration of thermal treatment produced recalcitrant and potentially toxic end products that reduced the biodegradability of wastewaters. Methane fermentation of the Maine-peat wet-carbonization waste was inhibited at an HRT of one day in the single-stage digester. However, two-phase digestion with separate acid-phase fermentation to promote hydrolytic degradation of toxic end products followed by separate methane fermentation at an HRT of 2 days exhibited a methane yield of 0.21 SCM/kg VS added and a BOD5 reduction of 76%. Volatile fatty acids profiles along the depths digesting cultures indicated that single-stage and two-phase digestion could be accomplished at one-half the reactor residence times (one day for single-stage and three days for two-phase) used in this research.
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Boyd, Ben J., and Andrew J. Clulow. "The influence of lipid digestion on the fate of orally administered drug delivery vehicles." Biochemical Society Transactions 49, no. 4 (August 25, 2021): 1749–61. http://dx.doi.org/10.1042/bst20210168.
Full textAbstract:
This review will focus on orally administered lipid-based drug delivery vehicles and specifically the influence of lipid digestion on the structure of the carrier lipids and their entrained drug cargoes. Digestion of the formulation lipids, which are typically apolar triglycerides, generates amphiphilic monoglycerides and fatty acids that can self-assemble into a diverse array of liquid crystalline structures. Tracking the dynamic changes in self-assembly of the lipid digestion products during digestion has recently been made possible using synchrotron-based small angle X-ray scattering. The influence of lipid chain length and degree of unsaturation on the resulting lipid structuring will be described in the context of the critical packing parameter theory. The chemical and structural transformation of the formulation lipids can also have a dramatic impact on the physical state of drugs co-administered with the formulation. It is often assumed that the best strategy for drug development is to maximise drug solubility in the undigested formulation lipids and to incorporate additives to maintain drug solubility during digestion. However, it is possible to improve drug absorption using lipid digestion in cases where the solubility of the dosed drug or one of its polymorphic forms is greater in the digested lipids. Three different fates for drugs administered with digestible lipid-based formulations will be discussed: (1) where the drug is more soluble in the undigested formulation lipids; (2) where the drug undergoes a polymorphic transformation during lipid digestion; and (3) where the drug is more soluble in the digested formulation lipids.
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KENNEDY, P. M., R. J. EARLY, R. J. CHRISTOPHERSON, and L. P. MILLIGAN. "NITROGEN TRANSFORMATIONS AND DUODENAL AMINO ACID CONTENT IN SHEEP GIVEN FOUR FORAGE DIETS AND EXPOSED TO WARM AND COLD AMBIENT TEMPERATURES." Canadian Journal of Animal Science 66, no. 4 (December 1, 1986): 951–57. http://dx.doi.org/10.4141/cjas86-105.
Full textAbstract:
Sixteen closely-shorn sheep fistulated at the rumen and duodenum were exposed to temperatures of 22–25 °C or 1–4 °C for periods of 45 d. Comparisons were made of digestion of nitrogen from alfalfa, red clover, reed canarygrass and bromegrass diets given in the chopped, or ground and pelleted form at intervals of 2 h. There was no consistent effect of environmental temperature on flow of nonammonia N to the intestines, irreversible loss rate of rumen ammonia, or rate of digestion of dietary N from nylon bags. In addition, apparent N digestibility in the gastrointestinal tract was not affected by environmental temperature, but cold exposure resulted in increases (P < 0.05) in apparent digestion of nonammonia N in the intestines relative to total intake of digestible organic matter. Grinding and pelleting of diets decreased (P < 0.001) rumen ammonia concentrations and increased (P < 0.01) total amino acid content of duodenal nonammonia N. There were dietary differences in flows of duodenal N, rumen ammonia concentration and rates of digestion from nylon bags. It was concluded that there were interactions between diet, dietary form, and the ratio of nitrogenous to energy-yielding metabolites which determine responses of ruminants to cold exposure. Key words: Cold stress, nitrogen, amino acids, sheep
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Navarro-Guillén, Carmen, Gabriella do Vale Pereira, André Lopes, Rita Colen, and Sofia Engrola. "Egg nutritional modulation with amino acids improved performance in zebrafish larvae." PLOS ONE 16, no. 4 (April 9, 2021): e0248356. http://dx.doi.org/10.1371/journal.pone.0248356.
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New and more efficient methods to sustainably intensify Aquaculture production are essential to attain the seafood demand for direct human consumption in the near future. Nutrition has been identified as one strategy of early exposure that might affect animal early development and later phenotype. This strategy may have positive consequences in the modulation of fish digestive physiology, which will correlate with higher performance outputs. Thus, improving fish digestive efficiency will lead to higher productivity and lower biogenic emission from aquaculture facilities, minimising the impact on the environment while increasing the biological efficiency. An innovative in ovo nutritional modulation technique based on low-frequency ultrasounds was used to enhance the transport of amino acids across the embryo membranes. An early stimulus with either arginine or glutamine, both involved in gut maturation, was applied in zebrafish (Danio rerio) embryos at 3.5 hours post-fertilization (hpf). At 22 days post-fertilization (dpf), growth performance, digestive enzyme activities and gut microbiota composition were analysed to evaluate the larval nutrition-induced metabolic plasticity and the effects on fish digestive efficiency. Results showed that fish survival was not affected either by the sonophoresis technique or amino acid supplementation. Final dry weight at 22 dpf was statistically higher in larvae from glutamine treatment when compared to the control even with lower trypsin activity, suggesting a higher nutrient digestion capacity, due to a slightly modulation of gut microbiota. Higher arginine supplementation levels should be tested as strategy to enhance growth at later developmental stages. In conclusion, this study demonstrated the efficiency of sonophoresis technique for in ovo nutritional modulation and suggests that in ovo glutamine supplementation might promote growth at later developmental stage through a positive microbiota modulation.
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Gu, Chunhe, Hafiz A. R. Suleria, Frank R. Dunshea, and Kate Howell. "Dietary Lipids Influence Bioaccessibility of Polyphenols from Black Carrots and Affect Microbial Diversity under Simulated Gastrointestinal Digestion." Antioxidants 9, no. 8 (August 17, 2020): 762. http://dx.doi.org/10.3390/antiox9080762.
Full textAbstract:
The bioaccessibility and activity of polyphenols is dependent on their structure and entrapment in the food matrix. While dietary lipids are known to transit into the colon, the impact of different lipids on the microbiome, and their interactions with dietary polyphenols are largely unknown. Here, we investigated the effect of dietary lipids on the bioaccessibility of polyphenols from purple/black carrots and adaptation of the gut microbiome in a simulated in vitro digestion-fermentation. Coconut oil, sunflower oil, and beef tallow were selected to represent common dietary sources of medium-chain fatty acids (MCFAs), long-chain polyunsaturated fatty acids (PUFAs), and long-chain polysaturated fatty acids (SFAs), respectively. All lipids promoted the bioaccessibility of both anthocyanins and phenolic acids during intestinal digestion with coconut oil exhibiting the greatest protection of anthocyanins. Similar trends were shown in antioxidant assays (2,2-Diphenyl-1-pricrylhydrazyl (DPPH), ferric reducing ability (FRAP), and total phenolic content (TPC)) with higher phytochemical bioactivities observed with the addition of dietary lipids. Most bioactive polyphenols were decomposed during colonic fermentation. Black carrot modulated diversity and composition of a simulated gut microbiome. Dramatic shifts in gut microbiome were caused by coconut oil. Inclusion of sunflower oil improved the production of butyrate, potentially due to the presence of PUFAs. The results show that the impact of polyphenols in the digestive tract should be considered in the context of other components of the diet, particularly lipids.
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Wang, Jing, Yong Chen, Xiaosong Hu, Fengqin Feng, Luyun Cai, and Fang Chen. "Assessing the Effects of Ginger Extract on Polyphenol Profiles and the Subsequent Impact on the Fecal Microbiota by Simulating Digestion and Fermentation In Vitro." Nutrients 12, no. 10 (October 19, 2020): 3194. http://dx.doi.org/10.3390/nu12103194.
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The beneficial effects of ginger polyphenols have been extensively reported. However, their metabolic characteristics and health effects on gut microbiota are poor understood. The purpose of this study was to investigate the digestion stability of ginger polyphenols and their prebiotic effects on gut microbiota by simulating digestion and fermentation in vitro. Following simulated digestion in vitro, 85% of the polyphenols were still detectable, and the main polyphenol constituents identified in ginger extract are 6-, 8-, and 10-gingerols and 6-shogaol in the digestive fluids. After batch fermentation, the changes in microbial populations were measured by 16S rRNA gene Illumina MiSeq sequencing. In mixed-culture fermentation with fecal inoculate, digested ginger extract (GE) significantly modulated the fecal microbiota structure and promoted the growth of some beneficial bacterial populations, such as Bifidobacterium and Enterococcus. Furthermore, incubation with GE could elevate the levels of short-chain fatty acids (SCFAs) accompanied by a decrease in the pH value. Additionally, the quantitative PCR results showed that 6-gingerol (6G), as the main polyphenol in GE, increased the abundance of Bifidobacterium significantly. Therefore, 6G is expected to be a potential prebiotic that improves human health by promoting gut health.
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Thandavhathu, M. R., Y. Zhao, G. Qin, Z. W. Shen, B. Wang, and Q. Zhang. "Effects of feed with different protein digestion kinetic profiles on intestinal health of growing pigs." South African Journal of Animal Science 51, no. 4 (September 16, 2021): 513–22. http://dx.doi.org/10.4314/sajas.v51i4.12.
Full textAbstract:
This study evaluated the effects of feed ingredients with different protein digestion kinetic profiles on the intestinal health of growing pigs. Two protein sources were selected, namely casein (CAS) as a rapid release source of amino acids (AAs), and corn gluten meal (CGM) as a slow-release source. Twenty-four crossbred barrows (Duroc × Landrace × Yorkshire) with similar bodyweight (43.27 ± 3.51 kg) were selected and randomly assigned to four treatments with six barrows. These consisted of T1: 13.2% digestible crude protein (CP) with supplemental CAS; T2: 13.2% digestible CP with supplemental CGM; T3: 11.2% digestible CP with supplemental CAS (T3); and T4: 11.2% digestible CP with supplemental CGM. Diets with CGM had increased crypt depth in the duodenum, jejunum, and ileum and reduced villi height in the jejunum in comparison with CAS. They also had increased intestinal permeability, as seen by the high level of serum diamine oxidase (DAO) compared with CAS. The diets with CAS increased health-promoting Lactobacillus and decreased health-threatening Treponema compared with those fed CGM diets. The CAS diets had a positive effect on gut functions with increased villi height, decreased crypt depth and high villi height/crypt depth. Thus, use of CAS in diets for pigs is favoured over CGM.
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Park, Nathan D., Ronald W. Thring, Randy P. Garton, Michael P. Rutherford, and Steve S. Helle. "Increased biogas production in a wastewater treatment plant by anaerobic co-digestion of fruit and vegetable waste and sewer sludge – A full scale study." Water Science and Technology 64, no. 9 (November 1, 2011): 1851–56. http://dx.doi.org/10.2166/wst.2011.753.
Full textAbstract:
Anaerobic digestion is a well established technology for the reduction of organic matter and stabilization of wastewater. Biogas, a mixture of methane and carbon dioxide, is produced as a useful by-product of the process. Current solid waste management at the city of Prince George is focused on disposal of waste and not on energy recovery. Co-digestion of fresh fruit and vegetable waste with sewer sludge can improve biogas yield by increasing the load of biodegradable material. A six week full-scale project co-digesting almost 15,000 kg of supermarket waste was completed. Average daily biogas production was found to be significantly higher than in previous years. Digester operation remained stable over the course of the study as indicated by the consistently low volatile acids-to-alkalinity ratio. Undigested organic material was visible in centrifuged sludge suggesting that the waste should have been added to the primary digester to prevent short circuiting and to increase the hydraulic retention time of the freshly added waste.
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Zhou, Xin, Zhao Zhang, Fenghong Huang, Chen Yang, and Qingde Huang. "In Vitro Digestion and Fermentation by Human Fecal Microbiota of Polysaccharides from Flaxseed." Molecules 25, no. 19 (September 23, 2020): 4354. http://dx.doi.org/10.3390/molecules25194354.
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The digestion of flaxseed polysaccharides (FSP) in simulated saliva, gastric and small intestine conditions was assessed, as well as in vitro fermentation of FSP by human gut microbiota. FSP was not degraded in the simulated digestive systems (there was no change in molecular weight or content of reducing sugars), indicating that ingested FSP would reach the large intestine intact. Changes in carbohydrate content, reducing sugars and culture pH suggested that FSP could be broken down and used by gut microbiota. FSP modulated the composition and structure of the gut microbiota by altering the Firmicutes/Bacteroidetes ratio and increasing the relative abundances of Prevotella, Phascolarctobacterium, Clostridium and Megamonas, which can degrade polysaccharides. Meanwhile, FSP fermentation increased the concentration of short-chain fatty acids, especially propionic and butyric acids. Our results indicate that FSP might be developed as a functional food that benefits gut health.
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Otálora, María Carolina, Robinson Camelo, Andrea Wilches-Torres, Agobardo Cárdenas-Chaparro, and Jovanny A. Gómez Castaño. "Encapsulation Effect on the In Vitro Bioaccessibility of Sacha Inchi Oil (Plukenetia volubilis L.) by Soft Capsules Composed of Gelatin and Cactus Mucilage Biopolymers." Polymers 12, no. 9 (September 2, 2020): 1995. http://dx.doi.org/10.3390/polym12091995.
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Sacha inchi (Plukenetia volubilis L.) seed oil is a rich source of polyunsaturated fatty acids (PUFAs) that are beneficial for human health, whose nutritional efficacy is limited because of its low water solubility and labile bioaccessibility (compositional integrity). In this work, the encapsulation effect, using blended softgels of gelatin (G) and cactus mucilage (CM) biopolymers, on the PUFAs’ bioaccessibility of P. volubilis seed oil was evaluated during in vitro simulated digestive processes (mouth, gastric, and intestinal). Gas chromatography–mass spectrometry (GC–MS) and gas chromatography with a flame ionization detector (GC–FID) were used for determining the chemical composition of P. volubilis seed oil both before and after in vitro digestion. The most abundant compounds in the undigested samples were α-linolenic, linoleic, and oleic acids with 59.23, 33.46, and 0.57 (g/100 g), respectively. The bioaccessibility of α-linolenic, linoleic, and oleic acid was found to be 1.70%, 1.46%, and 35.8%, respectively, along with the presence of some oxidation products. G/CM soft capsules are capable of limiting the in vitro bioaccessibility of PUFAs because of the low mucilage ratio in their matrix, which influences the enzymatic hydrolysis of gelatin, thus increasing the release of the polyunsaturated content during the simulated digestion.
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Nalle, C. L., V. Ravindran, and G. Ravindran. "Nutritional value of peas (Pisum sativum L.) for broilers: apparent metabolisable energy, apparent ileal amino acid digestibility and production performance." Animal Production Science 51, no. 2 (2011): 150. http://dx.doi.org/10.1071/an10100.
Full textAbstract:
Two experiments were conducted to evaluate the nutritional value of four cultivars (Santana, Miami, Courier and Rex) of peas (Pisum sativum L.) for broilers. In Experiment 1, the apparent metabolisable energy (AME) and the apparent ileal amino acid digestibility of these four cultivars were determined. The cultivar effects were found to be not significant (P > 0.05) for the AME and apparent ileal digestibility of amino acids, with the exception of arginine, which was lower (P < 0.05) in Courier than other cultivars. In Experiment 2, using the energy and digestible amino acid values determined in Experiment 1, diets containing 200 g/kg of the four cultivars of peas were formulated and the effects of feeding these diets on the performance and digestive tract development of broiler starters was investigated. Weight gain, feed intake and feed per gain of broiler starters fed diets containing peas were similar (P > 0.05) to those fed the maize-soybean meal diet. In general, the digestive tract development was unaffected (P > 0.05) by the inclusion of peas. The excreta scores of birds fed diets based on Santana, Miami and Rex were similar (P > 0.05) and that of the Courier-based diet was lower (P < 0.05) than those fed the maize-soy control diet. These results suggest that peas are good sources of metabolisable energy and digestible amino acids, and that they can be included at 200 g/kg level as a partial replacement for soybean meal in broiler starter diets without adverse effects on performance.
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Bosch, M. W., and P. van Leeuwen. "Studies to determine energy and protein requirements of the gastro intestinal tract of pigs." Proceedings of the British Society of Animal Production (1972) 1993 (March 1993): 44. http://dx.doi.org/10.1017/s0308229600023722.
Full textAbstract:
In intensive animal production, surplusses of nitrogen (N) are polluting the environment. Therefore it is important to compose diets matching the requirements of the animals as close as possible. Untill now mostly apparently digestible N or amino acids (feacal) have been used as criteria. It is also well established that amino acids disappearing from the hindgut do not contribute much to amino acid supply of the animal. Moreover, big differences between ileal digestible amino acids and faecal digestible amino acids are found, depending on the feed composition. Because the amounts of absorbed amino acids is more reflected by the amounts of ileal digestible amino acids than by the amounts of faecal digestible amino acids, contents of the ileal digestible amino acids of the commonly used feed stuffs in the Netherlands have been determined. Diets can now be formulated on contents of ileal digestible amino acids.
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Igual, M., P. García-Segovia, and J. Martínez-Monzó. "Amino acids and protein in vitro bio-accessibility from edible insect and pea protein enriched bread." Journal of Insects as Food and Feed 7, no. 6 (September 11, 2021): 1001–9. http://dx.doi.org/10.3920/jiff2020.0122.
Full textAbstract:
Edible insects have seen increasing use as a nutritive ingredient in food because of their high protein content and digestibility of amino acids from insect-enriched food. Therefore, the aim of this work was to evaluate amino acids and proteins bio-accessibility in bread enriched with Alphitobius diaperinus and Tenebrio molitor, comparing with a control and pea protein enriched bread. Bread was enriched at 5 and 10% with experimental ingredients. Experimental bread and the control were subjected to standardised static in vitro gastrointestinal digestion. Total amino acids (TAA) of breads were determined before digestion and free amino acids were determined after digestion by high-performance liquid chromatography. Crude protein was determined before and after gastrointestinal digestion. Incorporating pea protein, A. diaperinus, and T. molitor powder in bread formulations showed higher essential amino acids and non-essential amino acids content (12.170-16.274 mgTAA/100 g) than the control bread (10.843 mgTAA/100 g), with increasing amino acids content when concentration in bread increased. Bio-accessibility of each amino acid varied as a function of the bread formulation, however, after in vitro gastrointestinal digestion, experimental bread showed higher accessible amino acids and protein content than the control bread.
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Macelline, Shemil P., Peter V. Chrystal, Sonia Y. Liu, and Peter H. Selle. "The Dynamic Conversion of Dietary Protein and Amino Acids into Chicken-Meat Protein." Animals 11, no. 8 (August 3, 2021): 2288. http://dx.doi.org/10.3390/ani11082288.
Full textAbstract:
This review considers the conversion of dietary protein and amino acids into chicken-meat protein and seeks to identify strategies whereby this transition may be enhanced. Viable alternatives to soybean meal would be advantageous but the increasing availability of non-bound amino acids is providing the opportunity to develop reduced-crude protein (CP) diets, to promote the sustainability of the chicken-meat industry and is the focus of this review. Digestion of protein and intestinal uptakes of amino acids is critical to broiler growth performance. However, the transition of amino acids across enterocytes of the gut mucosa is complicated by their entry into either anabolic or catabolic pathways, which reduces their post-enteral availability. Both amino acids and glucose are catabolised in enterocytes to meet the energy needs of the gut. Therefore, starch and protein digestive dynamics and the possible manipulation of this ‘catabolic ratio’ assume importance. Finally, net deposition of protein in skeletal muscle is governed by the synchronised availability of amino acids and glucose at sites of protein deposition. There is a real need for more fundamental and applied research targeting areas where our knowledge is lacking relative to other animal species to enhance the conversion of dietary protein and amino acids into chicken-meat protein.
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Yoshioka, M., R. H. Erickson, J. F. Woodley, R. Gulli, D. Guan, and Y. S. Kim. "Role of rat intestinal brush-border membrane angiotensin-converting enzyme in dietary protein digestion." American Journal of Physiology-Gastrointestinal and Liver Physiology 253, no. 6 (December 1, 1987): G781—G786. http://dx.doi.org/10.1152/ajpgi.1987.253.6.g781.
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The role of rat intestinal angiotensin-converting enzyme (ACE; E.C 3.4.15.1) in the digestion and absorption of dietary protein was investigated. Enzyme activity was associated with the brush-border membrane fraction, with the highest activity in the proximal to midregion of the small intestine. Preliminary enzyme characterization studies were carried out using purified brush-border membrane preparations. When a variety of N-blocked synthetic peptides were used as potential substrates for ACE, activity was highest with those containing proline at the carboxy terminal position. The hydrolytic rates observed with these prolyl peptides were comparable to those observed when major digestive peptidases of the brush-border membrane such as aminopeptidase N and dipeptidyl aminopeptidase IV were assayed. When isolated rat jejunum was perfused in vivo with solutions of Bz-Gly-Ala-Pro, the dipeptide Ala-Pro was the main hydrolytic product detected in the perfusates. Absorption rates of the constituent amino acids, alanine and proline, depended on the concentration of peptide perfused. Captopril, an active site specific ACE inhibitor, significantly inhibited hydrolysis and absorption of constituent amino acids from Bz-Gly-Ala-Pro. These results show that intestinal brush-border membrane ACE functions as a digestive peptidase in addition to its role as a regulator of biologically active peptides in other tissues.
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Falavigna, C., M. Cirlini, G. Galaverna, and C. Dall'Asta. "Masked fumonisins in processed food: co-occurrence of hidden and bound forms and their stability under digestive conditions." World Mycotoxin Journal 5, no. 3 (August 1, 2012): 325–34. http://dx.doi.org/10.3920/wmj2012.1403.
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Fumonisins can occur in foodstuffs as free forms (prent, partially hydrolysed, or totally hydrolysed), as covalently bound fumonisins (bound fumonisins) and as non-covalently bound fumonisins (hidden fumonisins). The formation of several covalently bound fumonisin B1 conjugates upon reaction with sugars and/or amino acids has been reported in the literature so far, but occurrence data are still very poor. Since very little is known about the relevance of both hidden and bound fumonisins in processed products, the present study aimed to evaluate the occurrence of both masked forms in maize-based food products and to study their stability under digestive conditions. The behaviour of covalently bound fumonisin B1 conjugates upon in vitro digestion was evaluated, demonstrating their stability under these conditions. In addition, the co-occurrence of hidden and bound fumonisins in maize-based food products was investigated by application of both alkaline hydrolysis and simulated in vitro digestion assay. The experiments clearly showed that bound fumonisins were rather stable under the applied conditions, whereas hidden fumonisins released their parent forms.
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Kim, Jae Cheol, Bruce P. Mullan, Jung Min Heo, Christian F. Hansen, and John R. Pluske. "Decreasing dietary particle size of lupins increases apparent ileal amino acid digestibility and alters fermentation characteristics in the gastrointestinal tract of pigs." British Journal of Nutrition 102, no. 3 (January 23, 2009): 350–60. http://dx.doi.org/10.1017/s0007114508191231.
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Two experiments were conducted to examine the effects of lupin particle size on amino acid (AA) and energy digestibility and fermentation characteristics in pigs. Expt 1 examined the effects of lupin variety (cv. Tanjil and Mandelup) and lupin particle size (746, 888, 1099 and 1136 μm) on the total-tract apparent digestibility of dietary components in 63·5 (sd 7·28) kg pigs. While variety had no effect on total-tract apparent digestibility, decreasing particle size of lupins linearly increased total-tract apparent digestibility of crude protein of diets containing 350 g lupins per kg (P < 0·01). Expt 2 examined the effect of lupin particle size (567, 995, 1198, 1250 and 1304 μm) on digestion and fermentation characteristics in 29·8 (sd 2·9) kg pigs. Pigs were fed the respective diets ad libitum for the first 2 weeks and fed at three times maintenance energy level in the third week. Pigs were euthanised under sedation at 46·7 (sd 4·21) kg to collect digesta samples along the intestinal tract. Decreasing particle size increased apparent ileal and total-tract digestible N (P < 0·01) and the apparent and standardised ileal digestible AA content (P < 0·05– < 0·001) of lupins. Decreasing particle size of lupins linearly decreased the molar proportion of straight-chain volatile fatty acids (sum of acetic, propionic and butyric acids; VFAAPB), while branched-chain fatty acids (sum of valeric, caproic, isobutyric and isovaleric acids) were linearly increased (P < 0·001). The results demonstrated that particle size of lupins is a critical factor influencing nutrient, especially AA, utilisation efficiency and fermentation characteristics in the gastrointestinal tract of pigs.
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Kopec, Rachel E., Catherine Caris-Veyrat, Marion Nowicki, Beatrice Gleize, Michel Carail, and Patrick Borel. "Production of asymmetric oxidative metabolites of [13C]-β-carotene during digestion in the gastrointestinal lumen of healthy men." American Journal of Clinical Nutrition 108, no. 4 (August 22, 2018): 803–13. http://dx.doi.org/10.1093/ajcn/nqy183.
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Abstract Background Asymmetric β-apo-carotenoids (nonvitamin A–active metabolites) of provitamin A carotenoids have been observed in humans, but no study has investigated their formation during digestion. Objective The aim of this study was to follow the formation and absorption of asymmetric β-apo-carotenoids during digestion. Design Healthy men were intragastrically and intraduodenally intubated, and randomly assigned to consume a lipid-rich control meal (n = 3) or a lipid-rich test meal containing 20 mg [13C-10]-β-carotene (n = 7). Digesta samples were collected over 5 h, and blood collected over 7 h. The triglyceride-rich lipoprotein (TRL) fractions of plasma were also isolated. Lipophilic extracts of digesta, plasma, and TRL were analyzed via a high-performance liquid chromatography-tandem mass spectrometry method developed to identify [13C]-labeled β-apo-carotenals/carotenone, [13C]-β-apo-carotenols, and [13C]-β-apo-carotenoic acids. Results Relative to [13C]-β-carotene, [13C]-β-apo-carotenal levels remained ∼3 orders of magnitude lower throughout digestion (no [13C]-β-apo-carotenols, or [13C]-β-apo-carotenoic acids were observed). A mixed model determined relative influence of digesta type and time on digesta metabolite level. Increasing time significantly increased the model levels of digesta [13C]-β-apo-10′,12′,14′,15-carotenal and [13C]-β-apo-13-carotenone (P < 0.05) and trended toward decreased [13C]-β-apo-8′-carotenal (P = 0.0876). Gastric digesta were associated with a significantly higher level of [13C]-β-apo-8′-carotenal (P = 0.0289), and lower levels of [13C]-β-apo-12′,14′,15-carotenal (P < 0.05), relative to duodenal digesta. Anticipated retinoids, but no asymmetric [13C]-β-apo-carotenals, [13C]-β-apo-carotenols, or [13C]-β-apo-carotenoic acids, were observed in the blood or TRL samples. Conclusions β-Carotene appears to be robust to digestion, with minor amounts of β-apo-carotenals/carotenone formed. Absence of asymmetric [13C]-β-apo-carotenals in plasma and TRL suggests lack of absorption, levels below the limit of detection, lack of stability, or further conversion during the digestive process to as-yet unidentified products. Lack of asymmetric [13C]-β-apo-carotenals in plasma also suggests a lack of postprandial intestinal BCO2 activity in healthy humans. This trial was registered at clinicaltrials.gov as NCT03492593.
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Hiraoka, M., N. Takeda, S. Sakai, and A. Yasuda. "Highly Efficient Anaerobic Digestion with Thermal Pretreatment." Water Science and Technology 17, no. 4-5 (April 1, 1985): 529–39. http://dx.doi.org/10.2166/wst.1985.0157.
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The effectiveness of an anaerobic digestion process with thermal pretreatment at a comparatively low temperature was examined. The waste activated sludge was thermally pretreated in the process. A laboratory scale experiment and a pilot plant study were conducted. Changes in volatile acids concentration, in major constituents of sludge and in molecular size distribution were examined before and after thermal pretreatment and anaerobic digestion. Thermal pretreatment resulted in an increase of more than 30 percent in digestion gas production. The major substrates decomposed were fats in the pilot plant study. Glyceride fatty acids are decomposed with thermal pretreatment and produce volatile acids such as acetic and propionic acids. The volatile acids are directly utilized by anaerobes to produce methane. Organic matter such as fats and carbohydrates are depolymerized with thermal pretreatment and produce soluble intermediate compounds such as higher fatty acids. The intermediates are utilized in the anaerobic digestion process. These two major effects facilitate the methane production process.
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Souza, N. K. P., E. Detmann, D. S. Pina, S. C. Valadares Filho, C. B. Sampaio, A. C. Queiroz, and C. M. Veloso. "Evaluation of chromium concentration in cattle feces using different acid digestion and spectrophotometric quantification techniques." Arquivo Brasileiro de Medicina Veterinária e Zootecnia 65, no. 5 (October 2013): 1472–82. http://dx.doi.org/10.1590/s0102-09352013000500028.
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The objective of this work was to evaluate combinations between acid digestion techniques and spectrophotometric quantification to measure chromium concentration in cattle feces. Digestion techniques were evaluated based on the use of nitric and perchloric acids, sulfuric and perchloric acids, and phosphoric acid. The chromium quantification in the solutions was performed by colorimetry and by atomic absorption spectrophotometry (AAS). When AAS was used, the addition of calcium chloride to the solutions as a releasing agent was also evaluated. Several standard samples containing known chromium contents were produced (0, 2, 4, 6, 8 and 10g of chromium per kg of feces) using cattle feces obtained from three different animals to evaluate the accuracy of the different combinations of techniques. The accuracy was evaluated by adjusting a simple linear regression model of the estimated values on the actual values of chromium content in the standard samples. Regardless of the digestion technique, the chromium content estimates in the standard samples obtained by colorimetry were not accurate (P<0.05). Considering the AAS quantification, the digestion techniques based on nitric and perchloric acids and based on sulfuric and perchloric acids provided complete chromium recovery (P>0.05). The use of the digestion technique in phosphoric acid provided incomplete recovery of the fecal chromium (P<0.05). Subsequently, the digestion techniques in nitric and perchloric acids and digestion in sulfuric and perchloric acids, both evaluated by AAS, were compared using 84 cattle feces samples. The results indicate that these techniques provide similar (P>0.05) fecal chromium contents.
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Schutte, J. B., M. W. Bosch, J. de Jong, E. J. van Weerden, and F. Koch. "Factors affecting the requirement of dietary sulphur-containing amino acids of young pigs." Netherlands Journal of Agricultural Science 39, no. 2 (June 1, 1991): 91–101. http://dx.doi.org/10.18174/njas.v39i2.16544.
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In the first of 3 experiments, requirement for total and digestible sulphur amino acids (SAA) was estimated in 240 pigs, 14 to 40 kg. A diet with low digestible amino acids content (digestible sulphur amino acids (SAA) 0.37%) was supplemented with all essential amino acids except methionine and 0, 0.05, 0.1, 0.1 or 0.2% synthetic DL-methionine added, providing diets with 0.42, 0.47, 0.52 and 0.57% digestible SAA. A control diet contained 0.52% digestible SAA. At similar levels of digestible SAA, there was no difference in performance between the experimental and the control diet, showing that optimum performance can be obtained with a less digestible diet supplemented with essential amino acids. Optimum performance was at 0.52% digestible SAA content in the diet, corresponding to about 0.65% total SAA. In experiment 2, 360 pigs 15 to 40 kg, were used to study SAA requirement at sub-optimal (0.60%) and optimal (0.70%) dietary threonine levels. Increasing threonine from 0.60 to 0.70%, increased (P
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Baptissart, Marine, Aurelie Vega, Salwan Maqdasy, Françoise Caira, Silvère Baron, Jean-Marc A. Lobaccaro, and David H. Volle. "Bile acids: From digestion to cancers." Biochimie 95, no. 3 (March 2013): 504–17. http://dx.doi.org/10.1016/j.biochi.2012.06.022.
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Rha, Chan-Su, Hyunbin Seong, Young Sung Jung, Davin Jang, Jun-Gu Kwak, Dae-Ok Kim, and Nam Soo Han. "Stability and Fermentability of Green Tea Flavonols in In-Vitro-Simulated Gastrointestinal Digestion and Human Fecal Fermentation." International Journal of Molecular Sciences 20, no. 23 (November 24, 2019): 5890. http://dx.doi.org/10.3390/ijms20235890.
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Flavonols, the second most abundant flavonoids in green tea, exist mainly in the form of glycosides. Flavonols are known to have a variety of beneficial health effects; however, limited information is available on their fate in the digestive system. We investigated the digestive stability of flavonol aglycones and glycosides from green tea under simulated digestion and anaerobic human fecal fermentation. Green tea fractions rich in flavonol glycosides and aglycones, termed flavonol-glycoside-rich fraction (FLG) and flavonol-aglycone-rich fraction (FLA) hereafter, were obtained after treatment with cellulase and tannase, respectively. Kaempferol and its glycosides were found to be more stable in simulated gastric and intestinal fluids than the derivatives of quercetin and myricetin. Anaerobic human fecal fermentation with FLG and FLA increased the populations of Lactobacilli spp. and Bifidobacteria spp. and generated various organic acids, such as acetate, butyrate, propionate, and lactate, among which butyrate was produced in the highest amount. Our findings indicate that some stable polyphenols have higher bioaccessibilities in the gastrointestinal tract and that their health-modulating effects result from their interactions with microbes in the gut.
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Khorasani, G. R., W. C. Sauer, L. Ozimek, and J. J. Kennelly. "Digestion of soybean meal and canola meal protein and amino acids in the digestive tract of young ruminants." Journal of Animal Science 68, no. 10 (1990): 3421. http://dx.doi.org/10.2527/1990.68103421x.
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