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Analysis of avocado allergen (Prs a 1) IgE-binding peptides generated by simulated gastric fluid digestion.

BACKGROUND: Resistance to pepsin digestion has been claimed to be a characteristic of food allergens that can induce severe adverse reactions. Moreover, pepsin treatment is included in protocols to evaluate the potential allergenicity of transgenic foods. Allergenic plant class I chitinases, such as avocado Prs a 1, are the panallergens involved in the latex-fruit syndrome. Previous reports indicated their susceptibility to simulated gastric fluid (SGF) digestion. OBJECTIVE: We sought to evaluate the IgE-binding capacity and the in vivo reactivity of the SGF products of the avocado allergen Prs a 1. METHODS: Patients with a clinical history of latex-fruit allergy syndrome, a positive skin prick test (SPT) response to Prs a 1, and specific IgE to avocado were selected. Untreated and SGF-digested Prs a 1 samples were analyzed by means of IgE and IgG immunoblotting, IgE immunoblotting and ELISA-inhibition assays, and SPTs. Peptides from SGF-digested samples were fractionated by means of HPLC, characterized by N-terminal amino acid sequencing and matrix-assisted laser desorption/ionization analysis, and tested for in vivo reactivity with SPTs. RESULTS: Neither protein staining nor IgE immunoblotting with a pool of sera from allergic patients resulted in the detection of any band after SDS-PAGE separation of an SGF-digested sample of Prs a 1. However, this sample showed a similar inhibitory potency to that of untreated Prs a 1 in both immunoblot- and ELISA-inhibition assays (up to 70% inhibition of the IgE binding to crude avocado extract) and induced positive SPT responses in 5 of 8 allergic patients. Peptides from SGF-digested Prs a 1 were separated by means of HPLC, and 4 of them reached more than 50% inhibition values when using avocado extract as the solid phase in ELISA-inhibition assays. Reactive peptides were located both in the N-terminal hevein-like domain and in the catalytic domain of Prs a 1. Those corresponding to the hevein-like domain (approximately 5100 d) produced positive SPT responses in 5 of 8 allergic patients, whereas 2 peptides located in the catalytic domain (approximately 1400 and 2500 d) were reactive in 2 or 3 of the 8 patients. CONCLUSION: Prs a 1 was extensively degradated when subjected to SGF digestion. However, the resulting peptides, particularly those corresponding to the hevein-like domain, were clearly reactive both in vitro and in vivo.

Allergens↗

Incomplete digestion of codfish represents a risk factor for anaphylaxis in patients with allergy.

BACKGROUND: Fish represents one of the most important allergenic foods causing severe allergic reactions. Nevertheless, it has been shown that gastric digestion significantly reduces its allergenic capacity. OBJECTIVE: In this study, we assessed the absorption kinetics of fish proteins and investigated the clinical reactivity of patients with fish allergy to codfish digested at physiological or elevated gastric pH. METHODS: Healthy individuals were openly challenged with codfish and blood samples were evaluated by histamine release for absorbed fish allergens. Patients with allergy were recruited on the basis of previously diagnosed codfish allergy. Fish extracts were digested with gastric enzymes at pH 2.0 and 3.0 and used for histamine release, skin prick tests, and titrated double-blind placebo-controlled food challenges. RESULTS: Ingestion experiments in subjects without allergy revealed absorption of biologically active fish allergens only 10 minutes after ingestion with maximal serum levels after 1 to 2 hours. Incubation of fish proteins with digestive enzymes at pH 2.0 resulted in a fragmentation of the proteins leading to a reduced biological activity evidenced by a significantly smaller wheal reaction and reduced histamine release. Fish digested at pH 3.0 revealed comparable reactivity patterns as undigested extracts. Moreover, these test materials triggered reactions at 10-fold to 30-fold lower cumulated challenge doses in patients with allergy. CONCLUSION: Our data indicate the paramount importance of gastric digestion for fish allergens because the quantitatively significant absorption and elicitation of symptoms seemed to take place in the intestine. CLINICAL IMPLICATIONS: Hindered digestion puts patients with fish allergy at risk to develop severe allergic reactions at minute amounts of allergens.

Adult↗

A multi-laboratory evaluation of a common in vitro pepsin digestion assay protocol used in assessing the safety of novel proteins.

Rationale. Evaluation of the potential allergenicity of proteins derived from genetically modified foods has involved a weight of evidence approach that incorporates an evaluation of protein digestibility in pepsin. Currently, there is no standardized protocol to assess the digestibility of proteins using simulated gastric fluid. Potential variations in assay parameters include: pH, pepsin purity, pepsin to target protein ratio, target protein purity, and method of detection. The objective was to assess the digestibility of a common set of proteins in nine independent laboratories to determine the reproducibility of the assay when performed using a common protocol. Methods. A single lot of each test protein and pepsin was obtained and distributed to each laboratory. The test proteins consisted of Ara h 2 (a peanut conglutin-like protein), beta-lactoglobulin, bovine serum albumin, concanavalin A, horseradish peroxidase, ovalbumin, ovomucoid, phosphinothricin acetyltransferase, ribulose diphosphate carboxylase, and soybean trypsin inhibitor. A ratio of 10U of pepsin activity/microg test protein was selected for all tests (3:1 pepsin to protein, w:w). Digestions were performed at pH 1.2 and 2.0, with sampling at 0.5, 2, 5, 10, 20, 30, and 60min. Protein digestibility was assessed from stained gels following SDS-PAGE of digestion samples and controls. Results. Results were relatively consistent across laboratories for the full-length proteins. The identification of proteolytic fragments was less consistent, being affected by different fixation and staining methods. Overall, assay pH did not influence the time to disappearance of the full-length protein or protein fragments, however, results across laboratories were more consistent at pH 1.2 (91% agreement) than pH 2.0 (77%). Conclusions. These data demonstrate that this common protocol for evaluating the in vitro digestibility of proteins is reproducible and yields consistent results when performed using the same proteins at different laboratories.

Clinical Laboratory Techniques↗

Host hemolymph proteins and protein digestion in larval Habrobracon hebetor (Hymenoptera: braconidae).

Host plasma proteins and protein digestion in larval parasitoids were studied during trophic interactions of the ectoparasitoid Habrobracon hebetor Say (Hymenoptera: Braconidae), with a host, larvae of the Indianmeal moth, Plodia interpunctella Hübner (Lepidoptera: Pyralidae). We could detect no apparent differences in host hemolymph protein patterns up to 72 h after paralysation and/or parasitization by H. hebetor. A 190 kDa putative apolipophorin I present in host hemolymph could not be detected in the midguts of feeding H. hebetor larvae indicating that it is rapidly digested. The major 60 kDa storage proteins (putative hexamerins) in host hemolymph were detected in the parasitoid midgut and were completely digested 24 h after cessation of feeding and the beginning of cocoon formation. Host hemolymph had a pH of about 6.4. The pH optima of the midgut proteinases in the larval parasitoid were in the alkaline region, but midgut fluid in feeding parasitoid larvae was about pH 6. 8. Based on enzyme activity against selected artificial proteinase substrates including azocasein, N-alpha-benzoyl-L-Arg p-nitroanilide (BApNA), succinyl-Ala-Ala-Pro-Phe p-nitroanilide (SAAPFpNA), succinyl-Ala-Ala-Pro-Leu p-nitroanilide (SAAPLpNA), and inhibition by selected proteinase inhibitors, serine proteinases appear to be the predominant class of enzymes involved in protein digestion in the midguts of H. hebetor. There is also an active aminopeptidase (LpNA) associated with the microsomal fraction of midgut preparations. There was no evidence for preoral digestion or ingestion of proteinases from host hemolymph by the parasitoid larva. There was a very active BApNAase in the soluble fraction of midgut extracts. This activity increased on a per midgut basis up to 24 h after the beginning of cocoon formation but decreased rapidly by 48 h. Two major (P1 and P3) and several minor proteinases were detected in midgut extracts of H. hebetor analysed with gelatin zymograms. The apparent molecular mass of P1 varied from 95 to 49 kDa depending on protein loading. P3 had an apparent molecular mass of 39 kDa that was independent of protein loading. In summary, electrophoretic evidence indicates that host hemolymph protein patterns do not change significantly for at least 72 h after paralysation by H. hebetor. The role, if any, of envenomization in preventing breakdown of hemolymph proteins during this time remains to be determined. Because the predominant host hemolymph proteins, a putative apolipophorin I and the putative hexamerins, are readily digested by the serine proteinases present in the midguts of this parasitoid larva, these or similar proteins would provide an easily digested source of dietary amino acids that could be used for development of artificial diets for this beneficial insect.

Animals↗

Wax digestion by the lesser honeyguide Indicator minor.

Records of the lesser honeyguide feeding on beeswax in the wild are common. However, little is known about the ability of these birds to digest wax. We studied the wax preference, digestive efficiency, retention time and gut structure of the lesser honeyguide in the laboratory. Wax was found to be eaten daily by the lesser honeyguide. Birds ate significantly more new beeswax and foundation wax than old, black beeswax, but required an additional source of protein to maintain body mass. In the absence of other feed, the digestive efficiency for foundation wax was above 90%, with a long transit time of 256 min. Examination of the digestive tract showed no specialisation and no crop. The presence of digestive enzymes, including lipase, in the pancreas and small intestine and very few microbes in the digestive tract of the lesser honeyguide suggests that wax digestion occurs through a biochemical pathway with endogenous avian enzymes.

Animals↗

Comparative nutrient digestibility of arctic foxes (Alopex lagopus) on Svalbard and farm-raised blue foxes (Alopex lagopus).

Arctic foxes from Svalbard (n=4) and farmed blue foxes (n=4) was used in a digestibility experiment with a high-carbohydrate feed to add more information to the nutritional physiology of the arctic fox, and to compare its digestive capacity with that of the farmed blue fox. The arctic fox has a diet containing mainly protein and fat from mammals and birds, while farmed blue foxes have been exposed to an omnivorous dietary regime for more than 80 generations. The experiment showed in general no difference in digestive capacity for protein and fat between the foxes (P>0.05), but for carbohydrates, including starch and glucose, the blue fox revealed higher digestibility values. The superior digestive capacity for carbohydrates in blue fox might be a result of a long-term selection of animals digesting dietary carbohydrates more efficiently, or that an early age exposition to dietary carbohydrates has given permanent improvement of the carbohydrate digestion in the gut.

Amino Acids↗

The amino acid composition of human milk corrected for amino acid digestibility.

Human milk was collected from women in their 10th-14th weeks of lactation, and was analysed for amino acids. Corrections were made for losses of amino acids which were presumed to occur during acid hydrolysis, using a non-linear mathematical model that describes the simultaneous processes of amino acid yield and decay. The mean amino acid composition of the human milk was found to be similar to previously reported estimates, although the cysteine content of the human milk in the present study was 20% higher than the average literature estimate. True (corrected for endogenous amino acid excretions) ileal amino acid digestibility of human milk was determined using the 3-week-old piglet as a model animal for the human infant. The piglets were given either human milk (n 6) or a protein-free diet (n 6) for a 6 d experimental period. Cr2O3 was added as an indigestible marker, to both the human milk and protein-free diet. At the end of the experimental period the piglets were anaesthetized and samples of digesta removed from the terminal ileum of each piglet. After sampling the piglets were killed. Endogenous ileal excretions of amino acids were determined in piglets fed on the protein-free diet. The true digestibilities of total N and amino acid N were 88% and 95% respectively. The true ileal digestibility of the non-amino acid N fraction in human milk, when calculated by difference was only 50%. The true digestibility of the amino acids in human milk ranged from 81-101% with threonine (86%) being the least digestible essential amino acid. When the true ileal digestibility values were used to correct the amino acid composition of human milk, the pattern of digestible amino acids in human milk was different compared with the currently recommended pattern of amino acid requirements for the infant.

Amino Acids↗

Variation in cell wall composition among forage maize (Zea mays L.) inbred lines and its impact on digestibility: analysis of neutral detergent fiber composition by pyrolysis-gas chromatography-mass spectrometry.

Cell wall digestibility is an important determinant of forage quality, but the relationship between cell wall composition and digestibility is poorly understood. We analyzed the neutral detergent fiber (NDF) fraction of nine maize inbred lines and one brown midrib3 mutant with pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS). Among 29 pyrolysis fragments that were quantified, two carbohydrate-derived and six lignin-derived fragments showed statistically significant genetic variation. The pyrolysis products 4-vinyl phenol and 2,6-dimethoxy-4-vinyl phenol were negatively correlated with digestibility, whereas furfural and 3-(4-hydroxyphenyl)-3-oxopropanal showed a positive correlation with digestibility. Linear discriminant analysis of the pyrolysis data resulted in the resolution of groups of inbred lines with different digestibility properties based on their chemical composition. These analyses reveal that digestibility is governed by complex interactions between different cell wall compounds, but that several pyrolysis fragments can be used as markers to distinguish between maize lines with different digestibility.

Breeding↗

Sequential in vitro pepsin digestion of uncooked and cooked sorghum and maize samples.

An in vitro protein digestion study, using pepsin, was carried out in uncooked and cooked sorghum and maize flour samples. The digestibility values from the uncooked samples showed that sorghum presents digestibility values similar to those of maize. In the case of the cooked samples, it was found that a wet cooking procedure promotes a decrease in sorghum protein digestibility when compared to maize. Electrophoresis was used to follow the in vitro pepsin sequential digestion procedure, and infrared spectroscopy was applied to establish its efficiency. SDS-PAGE results showed that both uncooked samples (sorghum and maize) behave in a similar way. The wet cooking procedure increases the amount of high molecular weight aggregates and promotes the appearance of two nonreducible and nondigestible 45 and 47 kDa proteins. These two protein fractions are directly related to the loss of digestibility. It was also shown that in cooked sorghum the monomers (gamma-, alpha-, and beta-) are more resistant to digestion than the corresponding uncooked samples.

Digestion↗

Determinants of lactose digestion in the miniature pig.

Although lactose is an important nutrient in the diet of the infant and child, the factors contributing to its digestion have not been clarified adequately. We sought to determine the degree to which lactase activity and small intestinal transit explain lactose digestion, the average error (SEE) in estimating lactose digestion using these parameters, and the effect of age. We compared lactose digestion from both a 7% lactose-containing formula and a solution by determining lactose in ileostomy output in pig littermates at 10 days, 4 weeks, and 10 weeks of age. The entire small intestinal mucosa was assayed for lactase specific activity (micromol x min-1 x g protein-1), total activity (micromol x min-1), and whole-villus lactase activity. Transit time (min), and transit rate (cm/min) were measured. Meal type did not affect lactose digestion. Lactose digestion was explained best by lactase specific activity (formula, R2 = 0.73, SEE = 1.1; solution, R2 = 0.69, SEE = 1.0; P < 0.001). The next best parameter was total transit rate (formula, R2 = 0.69, SEE = 2.0; solution, R2 = 0.46, SEE = 1.3). The relationship with lactase specific activity was age related and there appeared to be a critical value of lactase specific activity above which essentially all the lactose was digested.

Aging↗

Adaptation to the digestion of nutrients of a starch diet or a non-starch polysaccharide diet in group-housed pregnant sows.

A trial was conducted with twenty group-housed pregnant sows to study the adaptation in nutrient digestibility to a starch-rich diet or a diet with a high level of fermentable non-starch polysaccharides (NSP) during a time period of 6 weeks. The starch-rich diet was primarily composed of wheat, peas and tapioca, whereas soya bean hulls and sugar beet pulp, which both are highly fermentable NSP sources, were used to formulate the NSP-rich diet. The starch-rich diet and the NSP-rich diet were formulated to contain different levels of starch (298 vs. 78 g/kg) and fermentable NSP (100 vs. 300 g/kg), but a similar level of net energy (NE) (8.36 MJ/kg). The trial consisted of a 1-week adaptation period followed by a 5-week collection period. Weekly apparent faecal digestibilities of dry matter (DM), crude protein (CP), crude fat, ash and NSP were measured by using the acid-insoluble ash marker method. Apparent faecal digestibilities of DM and organic matter (OM) of both diets were similar. Faecal digestibility of CP and crude fat was lower (p < 0.001) whereas that of NSP was higher (p < 0.001) for sows that received the NSP-rich diet. Calculated NE values of both diets were similar. Sows fed the NSP-rich diet produced faeces that contained a lower (p < 0.001) DM content compared with sows that were fed with the starch-rich diet. The quantity of dry faeces was the same on both diets, therefore total faeces production (as-is basis) was higher (p < 0.01) for the sows fed the NSP-rich diet. During the 5-week collection period, no changes were observed in the digestibility of DM, OM and NSP in the NE value of the diets. Digestibilities of CP and fat, however, were lower in week 1 (p < 0.05) compared with weeks 2-5 for both diets. The DM content of the faeces and the quantity of dry faeces did not change from weeks 1-5. Diet by time interaction was not observed for any of the response variables indicating that sows adapt as quickly to a diet with a high level of fermentable NSP as to a starch-rich diet. The present trial shows that, with regard to digestibility of nutrients, pregnant sows completely adapt to a NSP-rich diet (i.e., NSP from sugar beet pulp) in 2 weeks and that the time period necessary to adapt to a starch-rich diet or a diet with a high level of fermentable NSP is similar.

Adaptation, Physiological↗

Influence of age and body size on the digestibility of a dry expanded diet in dogs.

The objectives of the study were (i) to evaluate the effects of age and body size on the apparent digestibility of a dry expanded diet and (ii) to determine whether poor faecal quality could be related to lower nutrient utilization. Four breeds of dogs were used: six Miniature Poodles (MP), six Medium Schnauzers (MS), six Giant Schnauzers (GS) and six Great Danes (GD). Digestive trials were performed at 11, 21, 35 and 60 weeks of age. Faecal moisture and scoring were recorded at the same periods. Digestibility coefficients of all macronutrients increased significantly (p < 0.05) with age in all four breeds. Organic matter digestibility increased from 80.2 +/- 1.5 to 85.5 +/- 1.5% in MP; from 80.2 +/- 0.9 to 86.3 +/- 0.5% in MS; from 79.6 +/- 4.2 to 88.9 +/- 0.5% in GS and from 85.7 +/- 1.5 to 88.4 +/- 0.8% in GD. Crude protein digestibility increased from 74.5 +/- 4.0 to 81.5 +/- 1.9% in MP; from 74.3 +/- 3.9 to 82.6 +/- 0.9% in MS; from 71.8 +/- 6.0 to 85.9 +/- 0.9% in GS and from 80.8 +/- 4.6 to 84.8 +/- 1.3% in GD between 11 and 60 weeks of age, respectively. Mean faecal moisture also increased over the growth, except in GS. Nutrient digestibility was significantly higher in large dogs at all age periods. However, larger dogs showed lower faecal scores and increased faecal moisture. In conclusion, these results show (i) an effect of age and body size on nutrients digestibility and (ii) that the lower faecal quality observed in healthy large breed dogs does not appear related to lower nutrient utilization.

Aging↗

A highly digestible sorghum mutant cultivar exhibits a unique folded structure of endosperm protein bodies.

The endosperm of a sorghum mutant cultivar, with high in vitro uncooked and cooked protein digestibilities, was examined by transmission electron microscopy and alpha-, beta-, and gamma-kafirins (storage proteins) were localized within its protein bodies. Transmission electron microscopy micrographs revealed that these protein bodies had a unique microstructure related to high protein digestibility. They were irregular in shape and had numerous invaginations, often reaching to the central area of the protein body. Protein bodies from normal cultivars, such as P721N studied here, with much lower uncooked and cooked digestibilities are spherical and contain no invaginations. Immunocytochemistry results showed that the relative location of alpha- and beta-kafirins within the protein bodies of the highly digestible genotype were similar to the normal cultivar, P721N. Gamma-kafirin, however, was concentrated in dark-staining regions at the base of the folds instead of at the protein body periphery, as is typical of normal cultivars. The resulting easy accessibility of digestive enzymes to alpha-kafirin, the major storage protein, in addition to the increased surface area of the protein bodies of the highly digestible cultivar appear to account for its high in vitro protein digestibility.

Cooking↗

Fertilizer calcium as a factor affecting the voluntary intake, digestibility and retention time of pangola grass (Digitaria decumbens) by sheep.

1. Pangola grass (Digitaria decumbens) grown with and without calcium fertilizer was cut at three stages of regrowth to measure voluntary intake of dry matter (DM) and digestibility of various components of the dried-grass diet by sheep kept in metabolism crates. To determine the extent of a simple Ca deficiency half the sheep on each diet was supplemented with 1-4 g Ca/d. Retention times of the various dietary components in the reticulo-rumen were also determined. 2. Feeding a Ca supplement had no effect on voluntary intake or digestibility. 3. Ca fertilizer increased the Ca content of the grass from 2-2 to 3-8 g/kg DM and DM digestibility from 0-455 to 0-476 (P less than 0-01) due to an increase in the digestibility of the hemicellulose. 4. Voluntary intake was increased from 38-8 to 43-2 g/kg body-weight0.75 per d by Ca fertilizer due to an 18% reduction in the period of time the DM was retained in the reticulorumen. 5. Regressions relating voluntary intake to DM digestibility for the Ca-fertilized and control grass were significantly different (P less than 0-01). When compared at the same DM digestibility the voluntary intake of the Ca-fertilized grass was 2-6 g/kg body-weight0.75 per d higher than that of the control. 6. It was concluded that Ca fertilizer increased both DM digestibility and voluntary intake as a result of changes in the structural composition of the grass and not by a simple increase in the Ca content of the diet.

Animal Feed↗

Assessment of the nutritional quality of proteins: the use of "ileal" digestibilities of amino acids as measures of their availabilities.

1. A comparative study was made of two biological techniques for assessing protein quality in wheat and barley, and in a soya-bean-protein isolate both as received, and after being heat damaged by autoclaving. Amino acid digestibility was determined by the "ileal" analysis technique while amino acid availability was measured in growth assays. Some chemical and microbiological tests were also done. 2. Heat treatment of the soya-bean-protein isolate caused little change in amino acid composition but the digestibility of all amino acids and the availability of lysine and methionine were severely reduced, lysine being most affected. The reduced availability of lsyine was not entirely attributable to impaired digestibility. 3. The amino acids in wheat and barley were highly digestible. The availability of methionine and lysine in barley and of methionine in wheat was high, whereas the availability of lysine in wheat was apparently much lower. 4. The results showed that for methionine, digestibility values are a fair measure of the availability. In heat-damaged soya-bean-protein isolate however, digestibility of lysine over-estimated the availability, indicating that amino acid digestibility may sometimes provide a misleading indication of nutritional value.

Amino Acids↗

The role of condensed tannins in the nutritional value of Lotus pedunculatus for sheep. 2. Quantitative digestion of carbohydrates and proteins.

Primary growth vegetative Lotus pedunculatus containing 46 and 106 g/kg dry matter (DM) of total condensed tannin and 3 and 14 g/kg DM of free condensed tannin, was cut and fed fresh at hourly intervals (750 g DM/d) to sheep fitted with permanent cannulas into the rumen and duodenum. Low- and high-tannin lotus contained respectively 41.3 and 31.6 g total nitrogen/kg DM and 132 and 152 g lignin/kg DM. The two forms of lotus were similar in carbohydrate composition. Nutrient intake was recorded and faecal output measured by direct collection. Digesta flow to the duodenum was estimated by measuring dilution at the duodenum of inert ruthenium phenanthroline (Ru-P) and chromium-EDTA markers continuously infused into the rumen. Effects attributable to condensed tannins were assessed by comparing the digestion of the two diets, and by comparing the digestion of each with predicted values for non-tannin-containing fresh forages fed at similar intakes. Apparent digestibility of all nutrients measured was less for high- than for low-tannin lotus (P less than 0.01). The levels of cellulose digested ruminally and post- ruminally in both forms of lotus were similar to predicted values. However, less hemicellulose and readily fermentable carbohydrate (RFC; soluble carbohydrate + pectin) was digested in the rumen in sheep given both forms of lotus than would be predicted for non-tannin-containing fresh forage diets, but this was compensated for by greater post-ruminal absorption of both nutrients. Total N gains across the rumen (duodenal N flow--total N intake) were 1.8 and 10.5 g/d for low- and high-tannin lotus v. predicted losses of 3.7 and 2.1 g/d for non-tannin-containing fresh forages given at the same total N intakes. Post-ruminal digestion of non- amonia -N (NAN; proportion NAN flowing at duodenum) was 0.71 and 0.67 for low- and high-tannin lotus respectively v. 0.76 for comparable non-tannin-containing fresh forages. Energy absorbed as amino acids from the small intestine was calculated to be 0.29 of metabolizable energy for both forms of lotus, compared with 0.17 and 0.21 for perennial ryegrass and white clover. It was concluded that the presence of condensed tannins in lotus markedly increased post-ruminal NAN absorption compared with non-tannin-containing fresh forage diets, but depressed ruminal digestion of RFC and hemicellulose.

Animal Feed↗

Digestion of polysaccharides, protein and lipids by adult cockerels fed on diets containing a pectic cell-wall material from white lupin (Lupinus albus L.) cotyledon.

1. The cell-wall material of white lupin (Lupinus albus L.) cotyledon is characterized by low contents of cellulose (47 g/kg) and lignin (17 g/kg) and a high content of pectic substances (710 g/kg). The digestion of lupin cell-wall material by adult cockerels was estimated using gas-liquid chromatographic analyses of alditol acetates derived from polysaccharide sugars. The analyses were performed in the destarched water-insoluble fractions of feed and excreta. Digestibility measurements were carried out using a 3 d balance period including a 2 d feeding period and a 24 h final starvation period. 2. In the first experiment, six animals were given a diet containing 510 g white lupin cotyledon flour/kg which was the only source of protein and cell walls in the diet. The apparent digestibility of cell-wall components was near zero. 3. In the second experiment, three diets were prepared by diluting a fibre-free basal diet (diet A) by a semi-purified cell-wall preparation introduced at two different levels: 100 g/kg (diet B) and 200 g/kg (diet C). The semi-purified cell walls were prepared from the white lupin cotyledon flour used in the first experiment. The true digestibilities of polysaccharides measured in birds given diets B and C were near zero. It is suggested that the measurement of the neutral-detergent fibre (NDF) content according to Van Soest & Wine (1967) is not a suitable procedure for estimating the undigestible fibre content in poultry nutrition as the cell-wall pectic substances are not included in the NDF measurement. 4. Addition of the semi-purified cell-wall preparation (Expt 2) resulted in a slight decrease in the apparent protein digestibility. This decrease might be explained by the addition of undigestible cell-wall protein. 5. Addition of the semi-purified cell-wall preparation (Expt 2) had no effect on the apparent lipid digestibility. 6. The metabolizable energy values of the basal diet fraction of diets B and C were calculated assuming that the added plant cell-wall fraction was of no energy value. These calculated values were similar to the measured metabolizable energy value of diet A (basal). Thus, the tested pectic plant cell walls seemed to act as a diluent. It is suggested that, on the point of the digestion yields, all types of plant cell walls would act as diluents, in poultry.

Animal Feed↗

The effect of condensed tannins on the site of digestion of amino acids and other nutrients in sheep fed on Lotus corniculatus L.

Sheep were used to evaluate the nutritional consequences of a low condensed-tannin concentration (22 g/kg dry matter (DM)) in lotus (Lotus corniculatus L.) (control group) compared with lotus given to sheep receiving intraruminal polyethylene glycol (PEG) infusion (PEG group). PEG selectively binds to tannins and prevents tannins from binding proteins. DM intakes (1430 (SE 28) g/d) and digestibility of energy (663 (SE 4.5) kJ/MJ intake) were similar for both groups but the apparent digestion of nitrogen was lower in the control sheep (0.70) than in the PEG sheep (0.78; P less than 0.001). The proportion of N apparently digested before the abomasum (i.e. in the rumen) was lower (P less than 0.05) in control sheep (0.12) than in PEG sheep (0.21; P less than 0.05). Rumen ammonia concentrations were lower (P less than 0.001) in control sheep than in PEG sheep. The proportion of neutral-detergent fibre (NDF) digested in the rumen was similar for both groups (0.48 (SE 0.012)) but less energy was digested in the rumen of the control (0.42) than of the PEG sheep (0.47; P less than 0.05). The flux of essential amino acids (EAA) through the abomasum of control sheep was 50% greater than that in PEG sheep; flux of non-essential amino acids (NEAA) was 14% higher in control than in PEG sheep. Apparent digestibility of EAA in the small intestine was similar for both treatments (0.67), but NEAA were less well digested in the control (0.55) than in the PEG sheep (0.69). The presence of tannins in the control group increased net apparent absorption of threonine (57%), valine (89%), isoleucine (94%), leucine (30%), tyrosine (41%), phenylalanine (93%), histidine (90%) and lysine (59%), and reduced NEAA absorption by 10%, compared with PEG sheep.

Abomasum↗