Milk lipids and neonatal fat digestion: relationship between fatty acid composition, endogenous and exogenous digestive enzymes and digestion of milk fat.
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AIMS: To measure the nutritive value of pasture in terms of digestible energy intake (DEI) and dry matter (DM) digestibility, and the effects of increased calcium (Ca) intakes on apparent mineral absorption and bone characteristics in grazing weanling Thoroughbreds. METHODS: DM intake (DMI) and DEI were determined in 16 weanling Thoroughbreds grazing pasture from their daily faecal DM output, measured over 8 days, divided by the DM indigestible fraction (1-digestible DM) determined in a 6-day digestibility trial. The DM, gross energy content, crude protein, soluble carbohydrate, acid detergent fibre, neutral detergent fibre, lipid, Ca, phosphorus (P), sodium (Na), potassium(K), sulphur (S) and magnesium (Mg) composition of perennial ryegrass/white clover pasture and faeces were determined and their digestibility and/or apparent absorption calculated. Calcium intake and bone growth studies used 17 weanlings, randomly divided into three groups and fed perennial ryegrass/white clover pasture and 0.5 kg grain, with or without a CaCO3 supplement, for 84 days. The animals in Group 1 (n=6) were on a low Ca diet (3.5 g/kg DM) and were fed pasture only; those in Group 2 (n=5) were on a medium Ca diet (6.3 g/kg DM);and those in Group 3 (n=6) were on a high Ca diet (12.0 g/kg DM). After 44 days the apparent absorption of Ca was determined from the differences between the Ca intakes and faecal Ca outputs. At periods just before and after Ca supplementation the horses were anaesthetised and the left radius, third metacarpus(Mc3) and first phalanx of the left foreleg were scanned using a peripheral quantitative computed tomography scanner to determine cortical mineral content, density, area, periosteal circumference and bone strength. To investigate gastrointestinal tract transit time and DM digestibility, five randomly selected horses were administered Swiss screen bags on several occasions via a nasogastric tube. Each 60 x 10mm bag contained 3.21 (SE 0.37) g of frozen minced grass. Bags were recovered from the freshly passed faeces and frozen until analysis. RESULTS: The DM digestibility of the pasture was 0.62, while the DMI and DEI of weanlings (300 kg, gaining 0.7 kg/day) were 5.5 kg/day and 63 MJ/day, respectively. Increasing Ca intake had no significant effect on DEI, DM digestibility or on the apparent absorption of Ca, P, Na and K, but decreased the apparent absorption of Mg from 0.50 to 0.38. Regardless of Ca intake, the apparent absorption of Ca was 0.56. During the Ca administration trial there were significant increases overtime in the bone strength (strain stress index) of the proximal phalanx, Mc3 and radial diaphysis. However, the increase in bone strength was not associated with increase in dietary Ca, as neither the medium- nor high-Ca intake groups differed significantly from the low-Ca controls. The mean transit time for the Swiss screen bags was 25.46 (SE 0.09) h, and transit times were similar whether the horses were grazing or confined in loose boxes, being 26.64 (SE 0.23) h and 24.33 (SE 0.13) h, respectively. The DM digestibility determined using the bags was 0.54, which was significantly lower than the 0.62 determined by direct faeces collection. CONCLUSIONS: Good growth rates were achieved in Thoroughbred weanlings grazing perennial ryegrass/white clover pasture containing 3.5 g Ca/kg DM and a DE of 11.4 MJ/kg DM. Increasing dietary Ca intake 3.5-fold for 3 months had negligible impact on bone growth and development.
The extensive pentachlorophenol (PCP) contamination and its increasing treatment costs motivate the search for a more competitive treatment alternative. In a municipal wastewater treatment plant, anaerobic sludge-handling processes comprises three bio-processes, namely the anaerobic sludge digestion, post-sludge digestion and sludge land application, which reduce sludge organic content and make sludge a good fertilizer for land application. Availability and effectiveness make the anaerobic sludge handling processes potential technologies to treat PCP-contaminated soil. The technical feasibility of using anaerobic sludge bioprocesses was studied by treating PCP soil in two pilot digesters to simulate the primary sludge digestion, in serum bottles to mimic the post-sludge digestion, and in glass pans to represent the on-site sludge application. For primary digestion, the results showed that up to 0.98 and 0.6 mM of chemical and soil PCP, respectively, were treated at nearly 100% and 97.5% efficiencies. The PCP was transformed 95% to 3-MCP, 4.5% to 3,4-DCP, and 0.5% to 3,5-DCP. For post-digestion, 100% pure chemical PCP and greater than 95% soil PCP were removed in less than 6 months with no chlorophenol residues of any kind. Complete removal of PCP by-products makes this process a good soil cleanup method. For on-site treatment, PCP was efficiently treated by multiple sludge application; however, the PCP residue was observed due to the high initial PCP content in soil. Overall, more mass PCP per unit sludge per day was processed using the primary sludge digestion than the on-site soil treatment or post-sludge digestion. And, sludge acclimation resulted in better PCP treatment efficiencies with all three processes.
The sizes of Hae III partial digestion products at D1S7, D2S44, D4S139, D5S110, D10S28, and D17S26 were evaluated in experimentally generated partial digestions of liquid blood DNA. The partial digestion products were highly predictable, suggesting a very high level of sequence conservation in regions flanking variable number tandem repeat (VNTR) blocks. Partial digestion bands associated with three-or-more-banded patterns were also characterized. Partial digestion of three-banded patterns can be used to determine whether the extra bands arise due to internal Hae III sites in the VNTR block and to identify hidden three-banded patterns. Partial digestion products from forensic casework also conformed to size expectations. Presumed partial digestion bands from 27 forensic samples were compared to the experimentally generated data. The causes of partial digestion are examined and recommendations for interpreting forensic DNA evidence exhibiting partial digestion products are given.
The aim of the experiment was to study the effects of 2 wheat cultivars (Baltimor and Scipion) with different hardness values (75 and 5, respectively) on 2 divergent lines (D+ and D-) of broiler chickens selected on the basis of their digestion ability assessed by AME(n). Wheat was incorporated at 54.6% in diets. The other main ingredients were soybean meal (35.3%) and rapeseed oil (5.5%). Diets were given as pellets from 7 to 26 d. The experimental design was a 2 x 2 factorial design testing 2 wheat cultivars (soft or hard) on 2 selected lines of broiler chickens (high AME(n) or low AME(n)). From 7 to 16 d, D+ line showed lower (P < 0.0001) feed intake and feed:gain ratio than the D- line. At 3 wk of age, the D+ chickens resulted in increased digestibility values (P < 0.01) and 9% increased AME(n) value (P < 0.0001) compared with D-. Wheat cultivar effects on feed efficiency and AME(n) differed between lines. In the D+ line, their values were about 6% higher (P < 0.05) with soft than with hard wheat, whereas they did not differ in the D- line. However, wheat cultivar effect on starch digestibility did not differ between lines; soft instead of hard wheat resulted in about 6% improvement (P < 0.0001) in both lines. In the D- line, soft instead of hard wheat tended to reduce lipid and protein digestibilities, which explained why the starch digestibility improvement due to soft wheat was not converted into a significant AME(n) improvement in D birds. Study of digestive organ size revealed that increased proventriculus and gizzard weight (P < 0.05) could be one of the causes for the better digestion capacity of the D+ line. The pancreas was bigger (P < 0.01) in D- than in D+ birds, which probably came from an adaptation to a digestive disorder in D- birds.
Mature ponies fitted with permanent ileal cannulas were used in a 3 X 3 Latin square experiment to quantify prececal, postileal and total tract digestion of hay protein. Coastal Bermuda grass (CB), containing 11.7% crude protein, and two qualities of alfalfa, containing 15.0% (low-protein alfalfa; LA) and 18.1% (high-protein alfalfa; HA) crude protein, were fed in coarsely chopped form at 2% of body weight daily. Total tract apparent digestibility of the N in HA (73.8%) was higher than that in CB (57.0%; P less than .05) and was slightly higher than that in LA (66.1%; P less than .10). Nitrogen in LA was apparently more digestible than that in CB (P less than .05). Apparent prececal digestibilities of N in LA and CB were 1.3% and 9.6%, respectively, and were lower (P less than .05), or tended to be lower (P less than .10), than the 21.0% observed for HA. In relative terms, an average of 9.4% of the total N digestion occurred in the upper tract when CB and LA were fed, whereas 28.5% of total N digestion occurred in the foregut when HA was fed. There was a slightly higher concentration of total plasma free amino acids (P less than .10) at 1 h postfeeding when horses were fed alfalfa. Also, N retention was higher when ponies were fed HA (P less than .05) than when LA or CB were fed. Apparent postileal N digestibility was 52.5% for CB, 65.7% for LA and 66.9% for HA. Differences were not significant, and the large intestine appeared to compensate for the inefficiency of N digestion in the upper tract.(ABSTRACT TRUNCATED AT 250 WORDS)
Selected compounds were used to study physiological processes associated with digestion in the western tarnished plant bug, Lygus hesperus Knight. Durations of passage and rates of absorption, digestion, and excretion were determined for a digestible protein (casein), a non-digestible protein (green fluorescent protein, GFP), and a non-digestible carbohydrate (dextran). Dextran was used as a control to monitor the non-absorptive flow rate of ingesta through the digestive system. Fluorescent tracking of FITC-conjugates of casein and dextran, as well as immunoblotting and immunofluorescent staining of casein and GFP, were used to monitor the degradation (in vitro) and ingestion, digestion, and distribution (in vivo) of the respective compounds. Under our experimental conditions, L. hesperus took discrete meals, feeding and excreting at 2-3 h intervals. Rate of food passage was variable. FITC-dextran was found in the fecal material of most insects by 6-8 h after treatment initiation; by 12 h, 95% of ingested FITC-dextran was recovered from all insects. FITC-casein was digested extensively in in vitro homogenates of gut, hemolymph, and salivary gland. In vivo, FITC-casein was ingested and partially absorbed as a holoprotein into the hemolymph. Ingested FITC-casein was partially degraded in the gut and hemolymph within 2 h of ingestion, and no holoprotein was found after 12 h. In contrast, there was no detectable degradation of GFP in hemolymph, gut, and salivary gland homogenates after 24 h of incubation. Ingested GFP was not degraded in gut or hemolymph up to 8 h after treatment initiation, but did transfer to the hemolymph as a holoprotein. Analysis of immunohistological images confirmed that GFP bound to gut epithelial cell brush-border membranes. However, the mechanism by which GFP and casein pass as holoproteins into the hemolymph remains unknown.
1. Three subsamples of a batch of meat and bone meal (MBM0) were heated at 150 degrees C for 1.5 h (MBM1.5), 3 h (MBM3) or 5 h (MBM5). 2. The MBM's were included in 4 wheat-gluten based diets, each limiting in lysine. 3. The lysine digestibility for each of the diets was determined using precision-fed intact adult cockerels and caecectomised adult cockerels, ileal cannulated adult cockerels, the ileal digesta and excreta of young growing chickens. 4. The results of the digestibility assays were compared with the relative amount of lysine available as determined by growth assay. 5. Comparison of the regression lines of digestible lysine (determined with the digestibility assays) on available lysine (determined by growth assay) indicated that while the magnitude of the digestibility values varied significantly (P less than 0.05) depending on the type of assay used, the differences in lysine digestibility as a result of heat treatment across the 4 diets was the same when comparing assays (the regression lines were parallel). 6. The results of each digestibility assay also correlated with relative digestibility values determined by growth.
The apparent precaecal crude protein digestibility and amino acid absorption as well as the total digestibility of the nutrients organic matter, crude protein, crude fat and crude carbohydrates were determined of rations consisting of barley plus oilmeals of two rapeseed varieties of double quality (DQ) from Canada (Canola) and the GDR as well as of one variety of single quality (SQ) with 10 female pigs weighing between 120 and 140 kg with ileorectal anastomoses (IRA) and 4 intact (INT) female pigs weighing between 150 and 160 kg. The three rapeseed meals (RSM) were furthermore fed to IRA pigs as sole protein source together with protein-free feed. Based on the digestibilities of the crude nutrients determined with INT pigs and calculated with the difference method the energy content of RSM DQ "Canola", DQ GDR and SQ GDR could be calculated as 495, 633 and 500 EFUpig/kg DM (average: 543 EFUpig). The precaecal digestibility of the crude protein of the above mentioned RSM sorts determined with IRA pigs and calculated with the difference method amounts to 73, 76 and 75% resp. and is thus lower than the average absorption of all amino acids. The precaecal digestibilities of crude protein and essential amino acids for the mixtures calculated from the digestibility values of the individual feedstuffs barley and the above mentioned RSM largely agree with the experimentally determined digestibility values, i.e. there is additivity. On an average of the values known from literature for DQ RSM (including our own results), the precaecal digestibility of crude protein amounts to 70, of lysine to 74, of methionine to 82, of cystine to 82, of threonine to 69, of tryptophan to 72 and of isoleucine to 74%.
BACKGROUND: The extent of fat emulsification affects the activity of digestive lipases in vitro and may govern digestion and absorption of dietary fat. OBJECTIVE: We investigated the effect of the fat globule size of 2 enteral emulsions on fat digestion and assimilation in humans. DESIGN: Healthy subjects received intragastrically a coarse (10 microm) and a fine (0.7 microm) lipid emulsion of identical composition in random order. Gastric and duodenal aspirates were collected throughout digestion to measure changes in fat droplet size, gastric and pancreatic lipase activities, and fat digestion. Blood lipids were measured postprandially for fat assimilation. RESULTS: Despite an increase in droplet size in the stomach (2.75-6.20 microm), the fine emulsion retained droplets of smaller size and its lipolysis was greater than that of the coarse emulsion (36.5% compared with 15.8%; P < 0.05). In the duodenum, lipolysis of the fine emulsion was on the whole higher (73.3% compared with 46.3%). The overall 0-7-h plasma and chylomicron responses given by the areas under the curve were not significantly different between the emulsions, but the triacylglycerol peak was delayed with the fine emulsion (3 h 56 min compared with 2 h 50 min). CONCLUSIONS: Fat emulsions behave differently in the digestive tract depending on their initial physicochemical properties. A lower initial fat droplet size facilitates fat digestion by gastric lipase in the stomach and duodenal lipolysis. Overall fat assimilation in healthy subjects is not affected by differences in initial droplet size because of efficient fat digestion by pancreatic lipase in the small intestine. Nevertheless, these new observations could be of interest in the enteral nutrition of subjects suffering from pancreatic insufficiency.
Although it is recognized that the potentially digestible fraction of substrates in the rumen is heterogeneous in composition, most deterministic models that describe ruminal in situ digestion assume existence of only one or two homogeneous fractions of potentially digestible material. Alternatively, a stochastic model based on heterogeneity of substrate has been developed, although its validity has not been determined. To evaluate such a model, alfalfa hay and wheat straw were physically fractionated by dry sieving. Dacron bags containing these fractions were incubated in the rumen of a steer and the residue that remained was analyzed sequentially for DM, NDF, ADF, ADL, and acid detergent insoluble ash. Digestion kinetics of chemical fractions were determined by simultaneous analysis of the digestion profiles of DM, NDF, ADF, ADL, and acid detergent insoluble ash. The weighted mean and variance of the fractional digestion rate constant were .083 h-1 and 4.7 x 10(-4) h-2 for alfalfa and .021 h-1 and 2.4 x 10(-5) h-2 for wheat straw, respectively. In contrast, fitting DM data to a stochastic model assuming heterogeneous rates, the mean and variance of the fractional digestion rate were .078 h-1 and 8.3 x 10(-11) h-2 for alfalfa and .018 h-1 and 5.3 x 10(-12) h-2 for wheat straw, respectively. Because the estimates of the variance approached zero for the stochastic model, it is implied that this model reduced to a model involving a single fractional digestion rate. It is concluded that parameter estimates obtained from a stochastic model do not necessarily have a biological meaning.
The objectives of this experiment were to determine dietary factors influencing the apparent digestibility of Mg by lactating dairy cows and to compare empirical apparent digestibility values with Mg absorption coefficients used in the dairy cattle nutrient requirement model of the National Research Council (NRC). Data were compiled from 8 experiments with 39 dietary treatments and 162 cows (all lactating Holsteins) in which apparent digestibility of Mg was measured using total collection of feces and urine. The concentration of dietary Mg ranged from 0.20 to 0.36% of DM (mean = 0.27%). On average, 19% of the dietary Mg came from a Mg supplement (MgO or MgSO4). The concentration of dietary K ranged from 1.07 to 2.65% (mean = 1.60%). The mean apparent digestibility of Mg was 0.18 and ranged from -0.04 to 0.33. The average digestibility was 30% lower than the mean value calculated by the NRC model. The primary reason for the low average Mg digestibility was high concentrations of dietary K. At a dietary K concentration of 1%, empirical data agreed with NRC estimates, but apparent Mg digestibility decreased 0.075 (+/- 0.035)/percentage unit of K in the diet. Lactating dairy cows had to consume an additional 18 g of Mg/d for every 1 percentage unit increase in dietary K above 1% to maintain the same intake of digestible Mg as that consumed when fed a diet with 1% K.
Dairy cows grazing high-digestibility pastures exhibit pronounced diurnal variation in ruminal pH, with pH being below values considered optimal for digestion. Using a dual-flow continuous culture system, the hypothesis that minimizing diurnal variation in pH would improve digestion of pasture when pH was low, but not at a higher pH, was tested. Four treatments were imposed, with pH either allowed to exhibit normal diurnal variation around an average pH of 6.1 or 5.6, or maintained at constant pH. Digesta samples were collected during the last 3 d of each of four, 9-d experimental periods. A constant pH at 5.6 compared with a constant pH of 6.1 reduced the digestibility of organic matter (OM), neutral detergent (NDF), and acid detergent fiber (ADF) by 7, 14, and 21%, respectively. When pH was allowed to vary (averaging 5.6), digestion of OM, NDF, and ADF were reduced by 15,30, and 36%, respectively, compared with pH varying at 6.1. There was little difference in digestion parameters when pH was either constant or varied with an average pH of 6.1. However, when average pH was 5.6, maintaining a constant pH significantly increased digestion of OM, NDF, and ADF by 5, 25, and 24% compared with a pH that exhibited normal diurnal variation. These in vitro results show that gains in digestibility and potential milk production can be made by minimizing diurnal variation in ruminal pH, but only when ruminal pH is low (5.6). However, larger gains in productivity can be achieved by increasing average daily ruminal pH from 5.6 to 6.1.
While surfactants are commonly used in preparing protein samples, their presence in a protein sample can potentially affect the enzymatic digestion process and the subsequent analysis of the resulting peptides by mass spectrometry. The extent of the tolerance of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) to surfactant interference in peptide analysis is very much dependent on the matrix/sample preparation method. In this work the effects of four commonly used surfactants, namely n-octyl glucoside (OG), Triton X-100 (TX-100), 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) and sodium dodecyl sulfate (SDS), for biological sample preparation on trypsin digestion and MALDI-MS of the resulting digest are examined in detail within the context of using a two-layer method for MALDI matrix/sample preparation. Non-ionic and mild surfactants, such as OG, TX-100 or CHAPS, are found to have no significant effect on trypsin digestion with surfactant concentrations up to 1%. However, TX-100 and CHAPS interfere with the subsequent peptide analysis by MALDI-MS and should be removed prior to peptide analysis. OG is an MS-friendly surfactant and no effect is observed for MALDI peptide analysis. The effect of SDS on trypsin digestion in terms of the number of peptides generated and the overall protein sequence coverage by these peptides is found to be protein dependent. The use of SDS to solubilize hydrophobic membrane proteins, followed by trypsin digestion in the presence of 0.1% SDS, results in a peptide mixture that can be analyzed directly by MALDI-MS. These peptides are shown to provide better sequence coverage compared with those obtained without the use of SDS in the case of bacteriorhodopsin, a very hydrophobic transmembrane protein. This work illustrates that MALDI-MS with the two-layer sample preparation method can be used for direct analysis of protein digests with no or minimum sample cleanup after proteins are digested in a solution containing surfactants.
A previously developed microwave heated vapor-phase digestion method for biological samples was modified to enable digestion of difficult to digest organic samples. Organic samples containing ca. 100 mg of organic carbon were digested using volume calibrated quartz inserts inside second generation type medium pressure microwave vessels. As digestion reagents, 98% sulfuric acid, 70% nitric acid and 30% hydrogen peroxide were used. The accuracy of the method was tested with six certified reference materials. Elements Ca, Fe, K, Na, Mg, P and Zn were determined from NIST-SRM 8433 corn bran. Elements Al, Fe, Cd, Cu, and Zn were determined from NRCC DOLT-2 dogfish liver. The element Cd was determined from IRMM-VDA Cd in polyethylene No. 001-004 reference materials. These elements were determined from digested samples by ICP-OES. The results were close or within certified limits. The modified method could digest nearly all the materials tested, including the above mentioned reference materials, 2-nitrobenzoic acid (2-NBA), 4-NBA and copper(II) phthalosyanine-3, 4',4',4'-tetrasulfonic acid tetrasodium salt (CPS). The method could not digest 3-NBA.
In the present study we examined carbohydrase activities during a complete 24-h cycle and during the first days of starvation in both adult and juvenile snails. The results indicated the predominant role of the digestive gland in the secretions of the enzymes responsible for degradation of most of the carbohydrates tested. Salivary glands secreted some digestive enzymes but in amounts lower than secreted by digestive gland. Enzymatic activities fluctuated during the first hours of digestion and also after the digestive tract was empty. The relatively high enzymatic activities recorded 24 h after the intake of food and during starvation could be due to the circadian rhythm of this species and/or to the participation of an existing microflora in the digestive tract of Helix lucorum. The double origin (exogenous and endogenous) of some digestive enzymes such as cellulases is discussed.
The digestibility of proteins and individual amino acids of nineteen selected foods was determined by an in vitro assay. Samples were hydrolysed with pepsin for 30 minutes in an acidic medium; the pH was then raised to 7.5 and the mixture poured into the dialysis bag (molecular weight cut-off 1000) of a digestion cell with pancreatin. Digestion products, mixtures of free amino acids and low molecular weight peptides which pass through the dialysis membrane, were collected for 6 hours by sodium phosphate buffer circulation. All proteins from animal sources displayed a digestibility similar to casein, except for breakfast sausage. Vegetable proteins showed intermediate digestibility, except for cereals (lower) or peanut butter (higher). Target amino acids of enzymes were generally more readily hydrolysed. However, compared to other animal proteins, glycine in milk products, valine, isoleucine, methionine and lysine in breakfast sausage and hot dog, and histidine in tuna were more easily released. Overheating of non-fat dried milk not only reduced the lysine digestibility, but also that of methionine, phenylalanine, histidine and cystine. Among vegetable proteins, wheat products were characterized by a relatively greater release of threonine, isoleucine and histidine, and peas by a lower digestibility of methionine and lysine. Proline of soy isolate and isoleucine of pinto bean were resistant to hydrolysis while arginine of pinto beans and of rice-wheat-gluten was easily released.