Alterations in the jejunum of young goats caused by feeding soy protein-based diets.

This study was designed to investigate whether soy protein or soy protein supplemented with indispensable amino acids (AA) change the protein expression pattern and utilization of pre-cursors for RNA biosynthesis in jejunal mucosa in relation to casein and whether these changes affect mucosal cell growth. Kids were fed comparable diets based on cow;s milk, of which 50% of crude protein were replaced by either casein (CAS), soy protein (SP) or soy protein supplemented with indispensible AA (SPA) for 34 days (n = 4/group). Jejunal tissue was collected 5 h after adding a single dose of (15)N-RNA to the diet, in order to determine morphology, protein repertoire by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry, and RNA biosynthesis by isotope ratio-mass spectrometry. In mid-jejunum, morphological alterations induced by partial replacement of casein with soy protein were accompanied by changes in mucosal proteins related to generation of the cytoskeleton and in pathways for mucosal RNA biosynthesis, resulting in a smaller re-utilization of dietary RNA pre-cursors and in an increased activity of enzymes involved in nucleic acid breakdown. Soy protein supplemented with indispensible aminoacids tended to revise mucosal growth retardation with no impact on salvage of dietary RNA pre-cursors for mucosal RNA biosynthesis, but changes in cytoskeleton generation. Feeding soy protein with supplementation of indispensible AA does not ameliorate soy protein effects on mucosal morphology and RNA metabolism in the jejunum in a significant manner.

A milk diet partly containing soy protein does not change growth but regulates jejunal proteins in young goats.

Soy protein is known to alter intestinal function and structure. We determined in young goats whether a diet partly containing soy protein differently affects intestinal morphology and the jejunal and hepatic proteome as compared with a milk diet. Fourteen male 2-wk-old White German dairy goat kids were fed comparable diets based on whole cow's milk in which 35% of the crude protein was casein (milk protein group; MP) or soy protein supplemented by indispensable AA (SPAA) for 34 d (n = 7/group). Body weight gain and food efficiency were not different. Jejunal and hepatic tissue was collected to determine intestinal morphology by microscopy and protein repertoire by 2-dimensional gel electrophoresis and mass spectrometry. Jejunal crypt depth was reduced and villus height to crypt depth ratio was higher in SPAA than in milk protein. Out of 131 proteins identified, 32 proteins were found to be differently expressed in both groups. In SPAA, down-regulated jejunal proteins were involved in processes related to cytoskeleton generation, protein, lipid, and energy metabolism. Downregulated hepatic proteins were related to glycolysis and Krebs cycle. Thirteen proteins were upregulated in SPAA. Among these, 2 hepatic proteins were related to carbohydrate breakdown. The other 11 jejunal proteins were involved in cytoskeleton assembly, proteolysis, and carbohydrate breakdown. In addition, glutathione-S-transferase was found to be upregulated in the medial jejunum. In conclusion, a SPAA diet as compared with a milk diet was related to changes in jejunal morphology and jejunal proteins relevant for protein turnover, energy metabolism, and cytoskeleton assembly with no apparent impact on animal BW gain.

Effect of a soy protein-based diet on ribonucleic acid metabolism in the small intestinal mucosa of goat kids.

This study was designed to investigate the effect of soy protein inclusion in milk replacer diets for goat kids on protein, RNA, and DNA contents in small intestinal mucosa, on the importance of RNA biosynthesis from dietary RNA precursors for mucosal RNA synthesis, and on the activities of enzymes involved in nucleotide degradation in small intestinal mucosa. Diets were based on cow's milk. In the control group, 35% of the milk protein was replaced by casein (CN) protein, and in the soy group (SPAA), the same amount of milk protein was replaced by soy protein supplemented with essential AA known to be at lower concentrations in soy than in CN (Thr, Val, Ile, Leu, His, Lys, Met). Diets were isonitrogenous and isoenergetic. At 47 d of age, goats were harvested and samples of proximal, middle, and distal jejunal mucosa were collected 5 h after feeding 15N-labeled RNA from yeast (13 mg/kg of body weight). Growth and feed conversion did not differ between the control and SPAA kids. Mucosal protein concentrations were lower in the SPAA than the control kids. Concentrations of RNA and DNA did not differ between feeding groups, but in all kids mucosal RNA concentrations were higher in proximal than in middle and distal jejunum. Protein:RNA ratios were higher in the control than the SPAA kids and were lowest in proximal jejunum. Activities of alkaline phosphatase in enterocytes were higher in proximal than in middle and distal jejunum. Activities of mucosal xanthine oxidase were highest in distal jejunum and were higher in the SPAA than the control kids, especially in the middle and distal sites. The 15N-enrichment of mucosal RNA was higher in the control than the SPAA kids, especially in distal jejunum, and was lowest in distal jejunum. In contrast, 15N-enrichment of urea in plasma tended to be higher and Gly concentration in plasma was lower in the SPAA than the control kids. Data indicate that protein content and the protein:RNA ratio were lower in jejunal mucosa of goat kids fed milk replacer with partial replacement of CN protein by soy protein. These findings were accompanied by a lower level of reutilization of preformed dietary RNA precursors for RNA biosynthesis in jejunal mucosa and a higher activity of xanthine oxidase. Thus, feeding soy protein instead of CN protein reduced the incorporation of preformed dietary RNA precursors for RNA biosynthesis in the mucosa and activated key enzymes involved in nucleic acid breakdown.

Measurements of heat production for estimation of maintenance energy requirements of Hereford steers.

This study aimed to test the hypothesis that maintenance energy requirement (MEm) can be estimated from continuous heat production measurements during a change from a near maintenance feeding level to far below maintenance for two consecutive days. The MEm of eight Hereford steers weighing 286 +/- 5 kg (mean +/- SE) was determined in a balance trial. In addition, during the 10-d collection period, the animals were kept in open-circuit respiration chambers to measure 24-h gas exchange continuously at 10-min intervals. During the balance trial, the animals were fed dried chopped grass twice daily at an estimated level of 1.2 x MEm. After termination of the collection period on the 11th d of the balance trial, the steers were offered 2 kg/d of wheat straw while only gas exchange was measured. Estimates of MEm were derived from heat production (HP) data. The analyses included values of 24-h HP, HP of the nocturnal period (0000 to 0630), HP of the nocturnal period (excluding HP caused by standing) during the grass-feeding period and 24-h HP, nocturnal HP, and nocturnal HP (excluding HP caused by standing) during the straw feeding period. The MEm predicted from estimates of HP measurements were 536 +/- 9, 470 +/- 8, 441 +/- 8, 435 +/- 8, 393 +/- 9, and 373 +/- 9 kJ.kg of BW(-0.75).d(-1), respectively, whereas MEm calculated from data of the balance trial were 416 +/- 9 kJ.kg of BW(-0.75).d(-1). Values predicted for nocturnal HP (excluding HP caused by standing) of grass fed animals, 24-h HP, and nocturnal HP during straw feeding did not differ significantly from MEm. The differences in MEm among animals were reflected by all estimates of HP, whereas the correlation with the 24-h HP during straw feeding reached 0.9 (P = 0.002). We conclude that the method described is adequate to determine MEm with a sufficient degree of accuracy.

Carbon and nitrogen content based estimation of the fat content of animal carcasses in various species.

The objective of this study was to explore whether the C and N content can be used to estimate the fat content of animal carcasses. Considering the mean C and N contents of body fat and body protein, the fat content (EE) [%] can be predicted from C and N values [%] according to the generally valid equation EE = 1.3038 x C - 4.237 N. The application of this equation to estimate the total fat content of all animal carcasses results in significant differences in fat content between predicted and measured values. Therefore, we derived specific equations for rats, pigs, cattle, sheep, broilers and mice to predict the fat content by dual linear regression analysis (y = EE [% DM], x1 = C [% DM], x2 = N [% DM]) based on measured fat, C, and N contents of animal body samples. The specific equations for different animals showed residual standard deviations of 1.55, 1.63, 1.12, 1.35, 1.85 and 0.92% fat for rats, pigs, cattle, sheep, broilers and mice, respectively.

Effect of a soy protein diet on protein and energy metabolism and organ development in protein-restricted growing pigs.

Two feeding experiments were carried out with castrated male pigs weighing between 10 and 30 kg to study acute and persisting dietary effects on growth and on protein and energy metabolism in growing pigs. Pigs were fed semi-synthetic isoenergetic and isonitrogenous diets at 50% protein requirement with either soy protein isolate (SPI) or casein (CAS) as sole protein source. Intake of protein and ME amounted to 9% w/w and 1800 kJ x kg BW (-0.62) x d(-1) in Exp. 1, respectively, and 9% w/w and 1430 kJ x kg BW(-0.62) x d(-1) in Exp. 2. The CAS diet was supplemented by Lys, Met, Thr and Trp. In Exp. 1 (acute effects), 18 pigs received the CAS diet for 24 days (period 1); 9 pigs were then switched to a SPI diet whereas 9 pigs continued on the CAS diet for another 31 days (period 2). In Exp. 2, a third period of 31 days was added in which the SPI group was switched back to CAS diet. The control group was fed on the CAS diet throughout Exp. 2 (86 days). Altogether the majority of parameters were not affected neither comparing SPI with CAS in Exp. 1 nor inspecting possible persistence of effects in Exp. 2. In detail, in Exp. 1 SPI compared to CAS feeding resulted in a lower efficiency of protein utilisation and lower protein retention. Attendant upon the lower protein retention an increased energy retention as fat was only observed in tendency. SPI feeding caused a decreased body weight, thyroid weight and increased hepatic carbohydrate content that persisted after the diet was changed back to CAS (Exp. 2).

Effects of protozoa on methane production in rumen and hindgut of calves around time of weaning.

Effects of the presence or absence of ciliate protozoa on methanogenesis in the rumen and hindgut were investigated in young calves during a 7-week period. Ten Holstein calves, aged 7 days, were divided in two groups (n = 5) and fed an increasing amount of a commercial milk replacer and small amounts of a calves starter. One group was inoculated with ciliate fauna on two occasions, week 5 and 6, while the second remained ciliate-free. The absence of protozoa in the rumen decreased rumen empty weight (-23%, P < 0.01), and rumen pool size of N (-36%, P < 0.01) and crude fat (-37%, P < 0.05). Rumen bacteria of non-faunated calves contained a higher proportion of total amino acid-N per 16 g N (+3%, P < 0.01) and D-alanine-N per 16 g N (+13%, P < 0.05) compared to faunated calves. Further results contain a reference for a higher bacterial mass in the ciliate-free rumen with an increased number of bacteria adherent to rumen mucosa. The CH4 production in the rumen increased exponentially with the increase in protozoa population size (R2 = 0.68). In presence of 46 x 10(4) protozoa per ml rumen fluid, the in vitro CH4 production of rumen fluid per mol total VFA was about 34% higher in faunated than in non-faunated calves (P < 0.001). Hydrogen (2H) recovery of rumen fermentation was positively correlated (R2 = 0.55) to the CH4 production rate. Methanogens were attached on rumen mucosa. Methanogenesis, induced by rumen mucosa attached bacteria, was stimulated by ruminal protozoa. In the absence of protozoa in the rumen, the acetate-propionate ratio and butyrate proportion of VFA were reduced. In vivo, in the absence of protozoa not only the whole animal CH4 production (-30%, P < 0.05) but also the digestibility of carbohydrates (-4%, P < 0.05) was reduced. Thereby no difference was observed in the intake of ME per kg DM between the groups. In conclusion, the methanogenesis in the rumen, but not in hindgut, is associated with the development of the ruminal protozoa population. The level of methanogenesis (mol/mol VFA) in the hindgut amounts to 20% of the ruminal methanogenesis.

Influence of nutrition level on digestibility in high yielding cows and effects on energy evaluation systems.

The objective of the study was to determine the effect of nutrition level (NL, multiples of maintenance energy requirement) on the digestibility of nutrients for dairy cows regarding the energy supply of the animal. The digestion of nutrients and energy was investigated in two trials using lactating dairy cows. The NL varied from 2.7 to 5.0 using diets similar composition. In addition, sheep were given the same feed with a NL of 1.4. Digestibility of dry matter (DM) and all specific measures of dietary components declined significantly as NL increased. Digestibility of energy decreased by 4.1% for each increase in NL. The metabolizable energy, the ability to metabolize energy (metabolizable energy/gross energy), and the content of net energy for lactation (NE(L)) per kilogram of DM intake were calculated for NL from 1 to 6 on the basis of these relationships and as well as the gross energy, methane energy, and urine energy. Accordingly the NEL content declined by 0.11 MJ/kg of DM intake or 1.6% as the NL increased by one unit. It means that the NE(L) requirement above the maintenance requirement increased by 0.07 MJ of NE(L) per kilogram of fat-corrected milk, if the NL increased by one unit.

Activation of skeletal muscle protein breakdown following consumption of soyabean protein in pigs.

Diets with protein of inferior quality may increase protein breakdown in skeletal muscle but the experimental results are inconsistent. To elucidate the relationship, pigs were fed isoenergetic and isonitrogenous diets based on soyabean-protein isolate or casein for 15 weeks, with four to six animals per group. A higher plasma level of urea (2.5-fold the casein group value, P = 0.01), higher urinary N excretion (2.1-fold the casein group value, P = 0.01), a postabsorptive rise in the plasma levels of urea, 3-methylhistidine and isoleucine in soyabean protein-fed pigs suggested recruitment of circulatory amino acids by protein breakdown in peripheral tissues. Significant differences between dietary groups were detected in lysosomal and ATP-dependent proteolytic activities in the semimembranosus muscle of food-deprived pigs. A higher concentration of cathepsin B protein was found, corresponding to a rise in the cathepsin B activity, in response to dietary soyabean protein. Muscle ATP-stimulated proteolytical activity was 1.6-fold the casein group value (P = 0.03). A transient rise in the level of cortisol (2.9-times the casein group value, P = 0.02) occurred in the postprandial phase only in the soyabean group. These data suggest that the inferior quality of dietary soyabean protein induces hormonally-mediated upregulation of muscle protein breakdown for recruitment of circulatory amino acids in a postabsorptive state.

Energy metabolism of cocks and broiler chickens fed on diets with different carbohydrate sources.

Energy balances of cocks and chickens were measured using the nitrogen-carbon-balance method. In Experiment 1 twelve adult White Leghorn cocks were fed alternately on a basal ration or on a supplemental ration composed of 75% basal diet and 25% carbohydrate source as a supplement. In Experiment 2 six groups of 12 male broiler chickens were fed successively on two diets each with different carbohydrate sources (40% of DM) and on two energy levels. The investigated carbohydrate sources were glucose, fructose, sucrose, maize starch, raw and steamed potato starch, dried sugar beet pulp, tapioca, wheat, maize, rye and barley. In both experiments the energy digestibility of the diets with raw potato starch, beet pulp and barley was significantly lower compared to the other diets. Digestibility of those ranged from 88 to 81%. By simple linear regression no significant differences in efficiency of utilisation of ME of the diets between the carbohydrate sources sugars, starches and cereal grains could be proved. The corresponding MEm values agreed very close among the diets (411 to 429 kJ.kg BW-0.75.d-1).

Influence of chlorocholinechloride-treated wheat on selected in vitro fertility parameters in male mice.

The aim of this study was to evaluate the influence of feeding with food and water containing chlorocholinechloride (CCC) on the fertility of male mice in a two-generation study. For this purpose the number of testicular spermatozoa and the relative proportion of primary and secondary spermatocytes involved in spermatogenesis were measured. Furthermore, the fertility of epididymal spermatozoa from tested male mice was investigated in a special in-vitro fertilization system. The experimental food was composed of CCC-treated wheat in the first experiment and CCC-free wheat and water mixed with pure CCC in the second experiment. The CCC residue content in the treated food and water was 0.21 mg/kg and 0.2 mg/L, respectively. Under the influence of feeding with CCC-treated wheat (Experiment 1) the fertilization and cleavage rates of oocytes incubated with spermatozoa from CCC-fed mice were reduced: the fertilization rate 65.1% vs. 21.1% and the cleavage rate 51.9% vs. 20.3%, p < 0.01 (control feeding vs. CCC feeding, respectively). Feeding of sperm donors with pure CCC mixed with untreated wheat pellets or water (Experiment 2) led to a reduction in the fertilization and cleavage rate (control: 60.8%, 32.4%; CCC-food: 29.8%, 12.1%; CCC-water: 30.1%, 10.2%; CCC-food/water: 36.6%, 12.5%; p < 0.01, respectively). The normal course of spermatogenesis was unchanged after the exposure to CCC. Testicular weight, the number of spermatozoa, and the proportion of haploid, diploid, and tetraploid testicular cells were not influenced. However, the functional competence of epididymal spermatozoa from CCC-fed donors was reduced, resulting in a significantly diminished fertilization and cleavage rate in vitro. The results suggest that CCC could interfere with epididymal protein secretion and the process of sperm maturation during passage through the epididymis.

Morphological and functional development of the rumen in the calf: influence of the time of weaning. 1. Morphological development of rumen mucosa.

The objective of this study was to determine whether the nutritional regimen of rearing calves would influence the morphometric and histological development of rumen mucosa. Twelve male Holstein calves 7 d of age were assigned to three groups of 4 animals each: milk group (I), early weaned (6 weeks) group (II) and late weaned (9 weeks) group (III). All animals received additional solid feed. Animals of group I were slaughtered after 6 weeks of age, whereas those in groups II and III were slaughtered after 9 weeks of age. At slaughter, the ruminal digesta amounted to 2035 g (milk group), 3092 g (late weaned group) and 5374 g (early weaned group). The differences in the ruminal molar percentage of SCFA were not significant. There was a trend for lower pH and higher SCFA concentrations in the order late weaned, early weaned and milk fed animals (pH: 6.4, 6.6 and 6.7, respectively; SCFA: 96, 87 and 77 mmol/l, respectively). The mean length (1.07 mm in milk group, 1.45 mm in late weaned group and 1.87 mm in early weaned group), width (0.43, 0.58 and 0.71 mm, respectively) and surface of papillae (190, 232 and 241 mm2/cm2 mucosa, respectively) increased with both the age of the animals and the elevated intake of solid feed, whereas the number of papillae (210, 140 and 92 per cm2 mucosa, respectively) decreased. In both milk-fed groups type A and B corneal cells were present in the Stratum corneum, whereas in the earlier weaned calves type C-cells could be also seen. These findings indicate a more advanced stage of development of the rumen epithelium in the earlier weaned calves fed higher amounts of concentrate and hay.

Influence of dietary concentrate to forage ratio on the development of rumen mucosa in calves.

Effects of structural and non-structural carbohydrates on the development of rumen fermentation and ruminal mucosa in calves were examined during the weaning period. Barley/soybean meal (SBM) group was fed a concentrate starting from 2 weeks of age, whereas alfalfa group received a mixture of concentrate and alfalfa hay in which the proportion of the latter was gradually increased from 20% to 70% between weeks 2 and 9 of age. The total volatile fatty acid concentration in rumen fluid of calves increased with age, but at 9 weeks there were no significant differences between the two diets (barley/SBM group 153 mmol/l, alfalfa group 150 mmol/l). Rumen papillae at 9 weeks of age, as compared to 6 weeks of age, were longer and fewer in number per square centimetre mucosa, with larger cut surface. This resulted in a higher surface of papillae per square centimetre mucosa at 9 weeks (barley/SBM group 286 mm2/cm2, alfalfa group 245 mm2/cm2) than at 6 weeks of age (barley/SBM group 217 mm2/cm2, alfalfa group 198 mm2/cm2). At 9 weeks of age, the pH (barley/SBM 5.0, alfalfa 5.7), the acetate to propionate ratio (barley/SBM 2.2, alfalfa 3.2) as well as the length of the papillae in the ventral ruminal sac (barley/SBM 1.96 mm, alfalfa 2.37 mm) were increased in the alfalfa group when compared to the barley/SBM group (P < 0.1). In the former group, the proportion of butyrate revealed significantly increased values at 4 and 6 weeks of age. In animals of the barley/SBM group at 9 weeks of age, characteristic protrusions with proliferated thick epithelium occurred on the papillae and increased the surface for absorption. On the epithelium (Stratum corneum) desquamating cells with parakeratosis could be observed. In the alfalfa group the papillae of the ventral ruminal sac were longer, without protrusions. The morphotypes of the adhering rumen microflora differed between the groups. It can be concluded that feeding greater amounts of non-structural carbohydrates increases the surface for absorption of the rumen epithelium in calves. The absence of hyperkeratosis and rumenitis in the barley/SBM group indicated that there is no reason to limit high starch diets in the early weaning period of calves.

The use of an in vitro cellulase enzyme technique for predicting the digestibility of tropical feeds and fodders.

An attempt has been made to find out if the organic matter digestibility of tropical feeds and fodders can be predicted from the results of an in vitro cellulase enzyme technique. The in vivo digestibility coefficient of 27 tropical feeds and fodders was determined in wethers. The enzyme insoluble organic matter content of these feeds was determined using a cellulase enzyme technique. The indigestible OM (g/kg feed DM) in vivo (y) was plotted against enzyme insoluble organic matter (g/kg DM) (x). The equation is y = 60 + 0.817x, R2 = 0.95; RSD = 32 g/kg DM. Equations for predicting DOM (g/kg DM) and digestibility of OM (%) are calculated. The testing of the equation on an independent data base of 40 individual feeds and fodders showed that the validity of the equation is fairly general.

Long-term selection for protein amount over 70 generations in mice.

Based on the outbred mouse strain Fzt: Du, which has been obtained by systematic crossing of four inbred and four outbred lines, a long-term selection experiment was carried out for total protein amount (PA) in the carcass, starting in 1975. An unselected control line (CO) was kept under the same management but without continuous protein analysis. The protein amount of male carcasses at 42 days of age (P42) increased from 2.9 g in generation 0 to 5.2 g at generation 70, representing 97% of a theoretical selection limit. The total selection response amounts to 2.3 g, which is about 80% above the initial value and corresponds to 9 sigma p or 12 sigma A. The estimated realized heritability of protein amount decreased from 0.56 to 0.03 at generation 70, which was due to an increase in phenotypic variance from 0.065 to 0.24 g2 and a reduction in genetic variance from 0.04 to 0.01 g2. Half the selection response was obtained after about 18 to 23 generations, a half-life of 0.25 to 0.3 Ne. The maximum selection response was 0.094 g/generation and the response was 0.01 g/generation at generation 70. The measurements of body weights at 0, 10, 21, 42 and 63 days throughout the experiment showed a strong correlated effect for all weights. The PA mice are one of the heaviest lines of mice ever reported, and do not differ significantly in their body composition from control mice at 42 days. The direct selection response was due primarily to increased general growth. Body weight and protein amount are phenotypically and genetically highly correlated (rp = 0.82, rA approximately 1); however, selection for body weight led to fatter animals, whereas selection for protein opposed increased fatness (at least until selection age). This may be of general importance in animal breeding. The comparatively high selection response in this experiment seems due to the heterogeneity of the base population, the relatively high effective population size, and the duration of the experiment.

[Results from measurements of nitrogen and energy metabolism in adaptive different cattle]. / Ergebnisse aus Stickstoff- und Energieumsatzmessungen an adaptiv differenten Rindern.

In experiments with Galloway (G), Highland (H) and Black-White Dairy (B) cattle no significant differences between the breeds were measured in the energy and nutrient digestibility and energy metabolizability of rations with high variation in the nutrient composition. In H and B cattle no differences existed in digestibility in relation to the environmental temperature (30, 18 and 3 degrees C). Lowering the environmental temperature from 18 to 12 and 4-6 degrees C resulted in no changes of heat production in G and H but in B heat production increased about 6% and 20% respectively.

[Nutrient composition of some newly bred high protein and/or high lysine grains and their digestibility determined on growing pigs]. / Nährstoffzusammensetzung und an wachsenden Schweinen ermittelte Verdaulichkeit einiger protein- und/oder lysinreicher Getreideneuzüchtungen.

With the view to complementing the feedstuff data store and, consequently, to further improving the GDR Feed Evaluation System, some newly bred high-protein and/or high-lysine cereal varieties and strains (spring barley, winter wheat, maize) were studied for their nutrient composition and digestibility. Apart from from WEENDER's feed analysis technique, more recent methods were applied to determine total fat (after HCl treatment), carbohydrates (enzymatic method), lignin and amino acids. The digestibility of the nutrients was determined using growing pigs of different live weight, the test rations being made up of the cereals under and supplementations of limiting amino acids as well as vitamins and minerals. In comparison with the values from currently applied tables, the newly bred strains and varieties proved to have markedly higher contents of crude protein, digestible crude protein, lysine and energetic feed equivalents. Compared to crude fat, the total fat values proved markedly higher in the barley and wheat samples. The readily soluble and easily hydrolizable carbohydrates found with the new analysis procedure suggested, were 100% digestible in all cereal samples used. Lignin proved the constituent most difficult to digest and must be regarded as virtually undigestible in the case of pigs.