Bioefficacy of L-lysine sulfate compared with feed-grade L-lysine-HCl in young pigs.

A pig growth assay was conducted to determine the relative biological value (RBV) of lysine from L-lysine sulfate compared with feed-grade L-lysine HCl. One hundred nursery pigs with an average initial BW of 9.5 +/- 1.5 kg were blocked by BW and gender and allotted randomly to five dietary treatments in five replicates of four pigs per pen. A corn-peanut meal diet containing 0.6% total lysine (as-fed basis) was supplemented with two levels (0.1 and 0.2%) of lysine from L-lysine-HCl or L-lysine sulfate. The RBV of L-lysine sulfate was determined using multiple regression slope-ratio methodology, with ADG and G:F as the response criteria. At the tested levels, linear responses for gain and G:F were obtained from increments of lysine from the two lysine sources. When ADG was regressed on supplemental lysine intake, the RBV of lysine in L-lysine sulfate was 99% of the RBV of lysine in L-lysine HCl. When G:F was regressed on supplemental lysine intake, the RBV of lysine in L-lysine sulfate was 97% of the RBV of lysine in L-lysine-HCl. The t-test analysis revealed that the RBV of lysine in L-lysine sulfate was not significantly different from the RBV of lysine in L-lysine HCl, which was assumed to be 100% bioavailable. In conclusion, L-lysine sulfate can replace L-lysine HCl in diets for growing swine.

Molecular ecological analysis of porcine ileal microbiota responses to antimicrobial growth promoters.

Cultivation-independent microbial molecular ecology approaches were used to examine the effects of antibiotic growth promoters on the pig ileal microbiota. Five-week-old barrows were fitted with a simple T-cannula at the distal ileum. Three diets meeting or exceeding the minimum nutrient requirements were fed for 5 wk and supplemented as follows: 1) negative control (no antibiotic; n = 5), 2) continuous tylosin administration (n = 5), and 3) an antibiotic rotation sequence (wk 1, chlorotetracycline sulfathiazole penicillin; wk 2, bacitracin and roxarsone; wk 3, lincomycin; wk 4, carbadox; wk 5, virginiamycin; n = 5). Ileal luminal contents were collected for DNA isolation at the end of each of the 5 wk of the testing period. The V3 region of 16S rDNA was amplified by PCR and analyzed via denaturing gradient gel electrophoresis (DGGE) and quantitative polymerase chain reaction (qPCR). Resulting PCR-DGGE band numbers (bacterial species) were counted, and the banding patterns analyzed by calculating Sorenson's pairwise similarity coefficients (C(S)), an index measuring bacterial species in common among samples. Band numbers and total bacterial DNA concentrations decreased (P < 0.05) temporally in antibiotic-treated pigs compared with controls. Comparisons between treatments yielded low intertreatment C(S) indices, indicating treatment-dependent alterations in banding patterns, whereas intratreatment comparisons revealed increased homogeneity in antibiotic-treated vs. control pigs. Sequence analysis of treatment-specific bands identified three Lactobacillus, one Streptococcus, and one Bacillus species that were diminished with antibiotic rotation treatment, whereas tylosin selected for the presence of L. gasseri. Lactobacillus-specific qPCR was performed and analyzed as a percentage of total bacteria to further evaluate the effects of antibiotic administration on this genus. Total bacteria were decreased (P < 0.05) by tylosin and rotation treatments, whereas the percentage of lactobacilli increased (P < 0.05) by d 14 and through d 28 in tylosin-treated pigs. The decrease in total bacteria by antibiotics may reduce host-related intestinal or immune responses, which would divert energy that could otherwise be used for growth. Conversely, the ability of tylosin to improve animal growth may relate to its apparent selection for lactobacilli, commensals known to competitively exclude potentially pathogenic species from colonizing the intestine.

Energy metabolism and protein balance in growing rats fed different levels of dietary fibre and protein.

A study was performed to investigate the effect of different levels of dietary fibre (DF) and dietary protein on visceral organ size, digestibility, nitrogen balance and energy metabolism in rats. Thirty-six male Wistar rats, initial body weight about 76 g were used in a factorial design consisting of three levels of DF (low, 100 g/kg DM: medium, 250 g/kg DM and high, 290 g/kg DM) and two levels of dietary protein (low, 120 g/kg DM and high, 223 g/kg DM). The added fibre source was soybean hulls and Danish fish meal was used as sole source of dietary protein. Measurements of gas-exchange were done on six rats (one group) while urine and faeces were collected individually. The ratio of food/empty body gain increased (P<0.05) with increasing DF and decreasing levels of dietary protein. The weight of the digestive tract was larger (P<0.05) in rats fed the high fibre diet than in those fed the low fibre diet. The digestibility of nutrients and energy decreased linearly with increasing level of soybean fibre (P<0.05). An increased intake of DF was associated with a concomitant loss of protein and energy to faeces. The microbial degradation of NSP and other unabsorbed carbohydrates caused considerably changes in N metabolism of the colon. In rats fed the low protein diets increased levels of DF decreased N excretion in urine and increased N excretion in faeces, while the ratio of retained/digested protein remained constant. When rats were fed the high protein diet protein retention dropped in response to DF both absolute and relative to digested amount, indicating that energy intake could be a limiting factor. Heat production as a percentage of metabolizable energy (HP/ME) was higher (P<0.05) in rats fed the low protein diet than in rats fed the high protein diet, but no significant difference was found among DF levels.

The influence of soy oligosaccharides on apparent and true ileal amino acid digestibilities and fecal consistency in growing pigs.

Fourteen ileally cannulated pigs (BW = 35 +/- 2 kg) were randomly allotted to a replicated 7 x 7 Latin square design experiment to evaluate the influence of the soybean oligosaccharides (OS), raffinose and stachyose, on ileal nutrient digestibility and fecal consistency. Semipurified diets containing soy protein concentrate (SPC) or soybean meal (SBM) as the sole protein sources were fed. Soy solubles (SS), a by-product of SBM processing containing 3.5% raffinose and 11.5% stachyose, were used to increase dietary raffinose and stachyose concentrations. The seven dietary treatments were SPC, SPC + 9% SS, SBM, SBM + 9% SS, SBM + 18% SS, SBM + 24,000 U alpha-galactosidase enzyme preparation/kg diet, and a low-protein casein (LPC) diet used to calculate true digestibility. Diets, with the exception of the LPC diet, were formulated to contain 17% CP. All diets contained 0.5% chromic oxide as a marker for ileal digestibility determination. The experimental periods were divided into a 5-d diet adaptation followed by 2-d of ileal digesta collection. Diets and digesta were analyzed for DM, N, Cr, amino acids (AA), raffinose, and stachyose. Fecal consistency was determined on d 6 and 7 of each experimental period. The apparent and true ileal AA digestibilities were not different (P < 0.05) for the SPC and SBM control diets. When SS was added to the SPC diet, apparent and true N and AA digestibilities were depressed (P < 0.05) with the exception of Trp and Pro. The apparent and true ileal N and AA digestibilities were not different (P > 0.05) between the SBM control and SBM + 9% SS diets with the exception of Glu. There was a linear decrease (P < 0.05) in apparent and true DM, Val, Gly, and Tyr digestibilities when increasing levels of SS were added to the SBM diet. The addition of alpha-galactosidase did not improve apparent or true ileal N or AA digestibilities except for apparent and true Val and Tyr. Ileal raffinose digestibility was improved (P < 0.05) by addition of a-galactosidase, but was not affected by any other dietary treatment. Ileal stachyose digestibility was not affected (P > 0.58) by treatment. Fecal consistency likewise was not affected (P > 0.36) by dietary treatment. In conclusion, soy OS reduced nutrient digestibilities, but the reductions were small, ranging from approximately 1.1 to 7.4 percentage units. This suggests that other factors may be negatively impacting SBM digestibility.

Regional and processor variation in the ileal digestible amino acid content of soybean meals measured in growing swine.

To assess differences in soybean meal quality related to region of production, researchers in Illinois, Kansas, North Carolina, The Netherlands, and Ohio collected four soybean meal samples processed locally at least 15 d apart. These samples were assayed for ileal amino acid digestibility by pigs using a common soybean meal and a soy protein concentrate as references, and a low-protein casein diet for determination of endogenous amino acid losses. Digestibility was determined at each university using seven barrows surgically fitted with ileal cannulas in a 7 x 7 Latin square design. The experimental diets contained 17% CP from the test material except for the low-protein casein diet. Animals were fed twice daily, 12 h apart, at a level of 45 g x kg(-0.75) BW for each meal. Following a 5-d adaptation period, ileal digesta were collected for two 12-h periods for 2 d to be used for determination of ileal digestibility. Variation in amino acid digestibility was very small among and within sites and was much smaller than variation in the concentration of amino acids. Among sites, samples from The Netherlands had less total and thus digestible lysine and methionine than the U.S. samples (P < 0.05). The soybean meals tested in this experiment were approximately 4% higher in amino acids than that reported in the NRC (1998). True (standardized) digestibilities, however, were very similar to NRC values except for cysteine and threonine, which were 5 and 3 percentage points lower in this experiment, respectively. In conclusion, soybeans grown in the United States and locally processed into soybean meal were very similar in nutritional composition. Soybean meals produced in The Netherlands were lower in lysine and methionine (P < 0.05) but had a digestibility similar to that produced in the United States.

Evaluation of the homoarginine technique for measuring true ileal amino acid digestibilities in pigs fed a barley-canola meal-based diet.

The homoarginine technique has been suggested as a means to determine true ileal amino acid digestibilities in nonruminant animals fed protein-containing diets. Conditions for guanidinating lysine to homoarginine in barley and canola meal and the effect of this process on nutrient composition and ileal digestibilities in the resulting material were investigated. Conditions tested were methylisourea concentration (0.4, 0.5, or 0.6 M) and reaction time (4 or 6 d) at pH 10.5. Using 0.4 methylisourea M solution for 4 or 6 d gave guanidination rates of 72.5 and 78.5% for barley and 72.3 and 75.2% for canola meal, respectively. Using 0.5 M gave 88.0 and 84.6% guanidination rates in barley and canola meal, respectively, after a 6-d reaction time. Under these conditions, guanidination did not change the nutrient composition of barley (P > 0.10), whereas it increased CP (38.4 vs 49.0%), crude fiber (10.2 vs 16.0%), acid detergent fiber (30.0 vs 43.4%) and neutral detergent fiber (29.8 vs 49.4%) levels in canola meal (P < 0.05). Four 33.6-kg barrows fitted with a simple T-cannula at the terminal ileum were fed a 16% CP unguanidinated barley and canola meal-based diet for four consecutive 14-d periods. Ileal digesta were collected continuously for 24 h on d 12 and 14 to determine apparent nutrient digestibilities. On the morning of d 14, pigs were fed a diet in which half of the barley and canola meal was replaced with guanidinated material for determining true ileal amino acid digestibilities. Digesta samples were pooled by pig and by 24-h period to give 16 observations per diet. Apparent ileal digestibilities of DM, CP, and AA in the unguanidinated and guanidinated barley-canola meal diet were similar (P > 0.10) despite the changes observed in canola meal. Apparent ileal lysine digestibility was 73.9 and 74.5% in the unguanidinated and guanidinated diet, respectively. The true ileal lysine digestibility was 88.1%. The present results show that guanidination does not interfere with digestion and further support the use of the homoarginine method for determining true ileal amino acid digestibilities in pigs fed practical diets. A methylisourea solution of 0.5 M and a 6-d reaction time are recommended for converting lysine to homoarginine in barley and canola meal.

Improved procedure and cannula design for simple-T cannulation at the distal ileum in growing pigs.

Inserting a device (cannula) into the ileum is a common technique for studying nutrient digestibility, determining the disappearance or synthesis of a component, live tissue sampling, and inserting compounds or therapeutic agents into a specific region of the gastrointestinal tract. The purpose of this study was to implement improved cannulation procedures and care and management practices for cannulated pigs. Using these improved procedures, we successfully cannulated and maintained 15 pigs, each with an initial body weight of 30 kg. The newly designed cannula afforded improved stability and ease of adjustment while providing increased comfort to the pigs. The cannula was exteriorized through the last two ribs, providing excellent stability and security. To increase pig comfort, the cannulation site was washed and dried thoroughly and ointment was applied to the skin twice daily throughout the 63-day experiment. These updated procedures and maintenance practices should be considered and implemented in future experiments involving intestinal cannulation in pigs.

Valine requirement of nursery pigs.

Six experiments were conducted to determine the true digestible valine requirement of 5- to 20-kg pigs. In Exp. 1, a valine-deficient diet for 5- to 10-kg pigs was developed and validated in terms of growth performance in response to supplemental L-valine. A different basal diet was validated for 10- to 20-kg pigs in Exp. 2. Both diets were demonstrated to be deficient in valine and to support performance equivalent to typical nursery diets when fortified with L-valine. In Exp. 3, true ileal digestibility of valine in the two basal diets was determined in eight pigs fitted with a simple T-cannula at the terminal ileum. Another four pigs received an enzymatically hydrolyzed casein-based diet to determine endogenous contributions to collected ileal digesta. The two diets were found to have true valine digestibilities of 82% (5- to 10-kg pigs) and 86% (10- to 20-kg pigs). In Exp. 4, 80 weaned pigs (5.8 kg) were offered the basal diet fortified with five incremental doses (0.08%) of L-valine. Weight gain increased quadratically (P < 0.05) with increasing levels of valine. Broken-line analysis revealed a true digestible valine requirement of 0.86 +/- 0.03%. In Exp. 5, the true digestible valine requirement of 10- to 20-kg pigs was estimated with 120 pigs (10.9 kg) using the second basal diet fortified with six incremental doses (0.05%) of L-valine. The data suggested a digestible valine requirement level of about 0.775%, which was reevaluated in Exp. 6, wherein pigs did not respond to levels of digestible valine higher than 0.775%. In conclusion, requirement estimates were 2.50 and 2.22 g of true digestible valine per megacalorie of ME for 5- to 10- and 10- to 20-kg pigs, respectively. These empirical estimates are in close agreement with recent estimates of the National Research Council Subcommittee on Swine Nutrition of 2.48 and 2.11 g of true digestible valine per megacalorie of ME, respectively.

The effect of feed intake on ileal rate of passage and apparent amino acid digestibility determined with or without correction factors in pigs.

The apparent ileal digestibilities of amino acids and rate of passage were evaluated in pigs (BW = 78.3 +/- 7.4 kg) fed a semipurified diet. The pigs were fed 1.82, 2.73, or 3.65 kg DMI/d. The highest level of feed intake was considered to be ad libitum feeding. The pigs were fed according to a 3 x 3 Latin square design and were allowed to adapt to each experimental diet for 5 d. This was followed by 1 d of continuous collection of ileal digesta and a 2nd d of continuous collection separated into six 2-h postprandial time blocks. Ytterbium chloride hexahydrate was used to determine rate of passage. The ileal digestibilities of amino acids and rate of passage were unaffected (P > 0.05) by level of feed intake. The use of correction factors to more accurately express amino acid concentrations in the diet and digesta affected (P < 0.05) the apparent ileal digestibility coefficients of some amino acids.

True ileal digestibility of amino acids in sow's milk for 17-day-old pigs.

The digestibility of amino acids in sow's milk consumed by young pigs is currently unknown because of difficulties associated with collecting an adequate quantity of milk, and also problems in cannulating suckling pigs. A total of 14 kg of sow's milk was collected, two soluble indigestible markers (Co-EDTA and YbC13) were added, and the milk was fed to four pigs at 17 d of age that were fitted with a simple T-cannula at the terminal ileum. Another four cannulated pigs were offered a similar amount of a 20% DM liquid diet based on enzymatically hydrolyzed casein and lactose to assess endogenous amino acid losses. All pigs were fed about 875 g of each diet per day in 10 hourly meals from 0700 to 1700. Following 2 d of adaptation, ileal digesta were collected from 0800 to 1800 for 2 d. Diets and digesta were analyzed for amino acids using appropriate hydrolysis and preoxidation procedures. Average nitrogen true digestibility was 88%, whereas amino acid true digestibilities ranged from 84% (cystine and threonine) to 100% (methionine, histidine, and glutamic acid); the average for all amino acids was 92 +/- 4%. Based on average values, true digestibility of essential amino acids was not different from that of nonessential amino acids (P > 0.10). In whole milk, amino acids found in abundance in whey proteins (i.e., cystine, glycine, and threonine) were less (P < 0.05) digestible than amino acids predominating in casein proteins (i.e., glutamic acid, proline, and methionine). When true ileal digestible amino acid concentrations in sow's milk were expressed as ratios to digestible lysine, it appeared that threonine, tryptophan, and arginine were lower than what might be considered optimal. In conclusion, amino acids in sow's milk were highly digestible, but most of the amino acids had true ileal digestibility values significantly less than 100%.

Effect of hydrolysis time on the determination of amino acids in samples of soybean products with ion-exchange chromatography or precolumn derivatization with phenyl isothiocyanate.

Accurate determination of amino acid levels in soy products facilitates optimum diet formulation and amino acid supplementation. A study was carried out to investigate the effect of hydrolysis time and method of amino acid measurement on amino acid levels. Correction factors to standardize amino acid levels to 24 h of hydrolysis were also determined. Six different soybean products were evaluated. Hydrolysis was carried out for 10 different periods of time. Amino acids were analyzed by both ion-exchange chromatography and precolumn derivatization with phenyl isothiocyanate. Both hydrolysis time and measurement method affected (P < 0.05) amino acid levels. Standard hydrolysis conditions (hydrolysis in 6 M HCl at 110 degrees C for 24 h) rarely provide the maximal amino acid values. Therefore, sequential hydrolyses curves were very useful and should be used.

Digestion of fat does not differ in growing pigs fed diets containing fish oil, rapeseed oil or coconut oil.

We studied the digestion of fat and fatty acids in diets containing oils with different fatty acid composition. Four barrows (initial weight 35 kg) were fitted with a simple T-cannula at the terminal ileum. Three wheat starch and fish meal-based diets were formulated to contain either 150 g fish oil, rapeseed oil or coconut oil/kg. A basal diet, which did not contain oil, was also prepared. The diets were fed according to a 4 x 4 Latin square design. Each experimental period comprised 5 d adaptation to the diets, 3 d fecal collection and 2 d digesta collection. The apparent ileal and fecal digestibilities of fat were relatively high (88 - 93%). The ileal digestibilities of total, saturated and monounsaturated fatty acids did not differ among the diets. However, the digestibilities of polyunsaturated fatty acids (PUFA) in the fish and rapeseed oil diets were higher (P < 0.05) than in the coconut oil diet. The ileal digestibilities of 18:1, 18:2 and 18:3 in the rapeseed oil diet ranged from 94 to 97%. The ileal digestion of the unsaturated long-chain fatty acids 20:5(n-3) and 22:6(n-3) in the fish oil diet was nearly complete (97 - 98%). Apparent fecal digestibilities of saturated fatty acids (76 - 89%) were lower than apparent ileal digestibilities (89 - 94%). The digestibilities of fat and fatty acids were relatively high when pigs were fed diets containing fish oil, rapeseed oil or coconut oil. There were few differences in the digestibilities of saturated, monounsaturated and PUFA in the fish oil, rapeseed oil or coconut oil diets.

Endogenous gut nitrogen losses in growing pigs are not caused by increased protein synthesis rates in the small intestine.

The purpose of this study was to establish, using a flooding dose of L-[ring 2, 6-(3)H] phenylalanine, whether feeding pigs diets that induce high endogenous gut nitrogen losses (ENL) also increases protein synthesis rates in (PSR) the visceral organs. Twelve 18-kg Yorkshire barrows with catheters in the right and left jugular veins were fed for 3 wk either casein-cornstarch- (CC) or barley-canola meal- (BCM) based diets formulated to a similar digestible energy /crude protein ratio and designed to induce either low or high ENL, respectively. Pigs were infused with 10 mL/kg body weight of a 150 mmol. L(-1) phenylalanine solution containing 230 MBq. L(-1) labeled phenylalanine for 12 min and killed 20 min later. Plasma phenylalanine specific radioactivity (SRA) rose to a plateau value within 3 min of starting the infusion and did not change (P > 0.10) thereafter. Fractional rates of protein synthesis (K(s), %/d) based on SRA in plasma- or intracellular-free phenylalanine did not differ (P > 0.10) in all tissues except pancreas (P < 0.05). Diet affected K(s )in liver (P < 0.01) and colon (P < 0.05) but not in pancreas, duodenum, jejunum and cecum. Based on plasma-free phenylalanine SRA, liver K(s)were 85.4 +/- 11.0 vs. 60.5 +/- 5.2 (mean +/- SEM) in CC- and BCM-fed pigs, respectively; these values were 82.3 +/- 4.7 vs. 98.2 +/- 5.8 in the colon. The absolute amount of protein synthesis (g.d(-1)) was higher in the liver (P < 0.05) and pancreas (P < 0. 05) of the CC pigs compared to BCM pigs. No dietary effects were observed in all other organs (P > 0.10). The present results suggest that feeding growing pigs a BCM diet that induces high ENL does not affect PSR in the small intestine of growing pigs from which >50% of ENL originates.

Pancreatic secretion of zinc and carboxypeptidase A and B in growing pigs.

The secretion of zinc and the level of carboxypeptidase A and B activity in pancreatic juice were studied in three pigs fitted with a pancreatic pouch re-entrant cannula (PM pigs) and three different pigs with a catheter surgically implanted in their pancreatic duct (CM pigs). The zinc in the pancreatic juice appeared to be primarily associated with carboxypeptidase A and B. Both the concentration of zinc in pancreatic juice and the daily secretion of zinc in PM pigs were greater than in CM pigs. However, compared to the daily intake of zinc, its secretion in pancreatic juice was low. The specific activity levels of carboxypeptidase A and B in pancreatic juice collected from PM pigs were higher than in CM pigs. The total activity of carboxypeptidase A in pancreatic juice did not differ between collection methods. The total activity of carboxypeptidase B was higher in pancreatic juice collected from PM than from CM pigs. The differences in zinc and carboxypeptidase secretion between PM and CM pigs were probably due to physiological changes induced following the different surgical preparations of the animals.

A comparison of two methods to measure amylase, lipase, trypsin, and chymotrypsin activity and the effect of freezing and thawing on enzyme activities in pancreatic juice.

An experiment was conducted to compare two methods, referred to as A and B, for measuring amylase, lipase, trypsin, and chymotrypsin activities in samples of pancreatic juice and to investigate the effect of freezing and thawing on enzyme activities. Three growing pigs were used in this study. A catheter was surgically inserted into the pancreatic duct and a simple T-cannula was placed in the duodenum to allow the return of collected pancreatic juice. Pancreatic juice was collected for a total of 24 h and 27 pooled 8-h samples were used in the analyses. Following collection and pooling, the samples were frozen at -80 degrees C, due to the large number of samples and analyses, and stored for 2 weeks. The samples were thawed and enzyme activities were determined with Method A. The samples were frozen again at -80 degrees C, stored for 2 weeks, and rethawed and enzyme activities were determined with Methods A and B. There was a direct relationship between Method A and Method B for measuring amylase, trypsin, and chymotrypsin activity (R2 = 0.95, 0.81, and 0.90, respectively). However, a poor relationship was observed between a titrimetric and a photometric method for determining lipase activity (R2 = 0.42). A second freezing, storage, and thawing did not affect (p > 0.3) the concentration of protein or amylase activity in pancreatic juice. However, the activities of chymotrypsin, trypsin, and, especially, lipase were decreased by 29.0% (p < 0.11), 40.4% (p < 0.01), and 82.9% (p < 0.001), respectively. In conclusion, either Method A or Method B can be used to measure amylase, trypsin, and chymotrypsin activities in pancreatic juice but a titrimetric method should be used to determine lipase activity. Furthermore, if the activities of lipase, trypsin, and chymotrypsin are to be determined, the sample should only be frozen and thawed once. If amylase activity will be determined, the sample can be repeatedly frozen and thawed.

Electrophoretic separation of proteolytic enzymes in pancreatic juice collected with the pouch or catheter method.

CONCLUSION: The results of this study demonstrated that proteolytic enzymes in pancreatic juice from pigs prepared with the pouch method (PM) were nearly fully active or were fully active. When activation with enterokinase was carried out further inactivation and/or breakdown occurred for chymotrypsin C and cathodal trypsin. In addition, some inactivation and/or breakdown of proteolytic enzymes in pancreatic juice occurred during collection of pancreatic juice from PM pigs. METHODS: Samples of pancreatic juice were collected from growing pigs using either the PM or the catheter method (CM). An isolated pouch was prepared where the pancreatic duct enters the duodenum, and three pigs were fitted with a pancreatic pouch re-entrant cannula. Three different pigs had a catheter surgically inserted into the pancreatic duct. Pooled 8-h samples of pancreatic juice were analyzed before and after activation with enterokinase. Chymotrypsin, trypsin, and elastase activities were identified in pancreatic juice after separation by electrophoresis in 1% agarose gels at pH 8.6 using N-acetyl-DL-phenylalanine-beta-naphthyl ester (Ac-Phe-beta ne) as a substrate. RESULTS: This qualitative enzyme assay indicated that a considerable amount of chymotrypsin C, anodal trypsin, chymotrypsins A and B, elastase II, and cathodal trypsin were present in samples of nonactivated pancreatic juice from PM pigs. In contrast, the only active enzymes identified in pancreatic juice from CM pigs were very small amounts of chymotrypsin A and elastase II. The amounts of chymotrypsin C and cathodal trypsin were lower in activated than in nonactivated pancreatic juice from PM pigs. However, there were increases in the amounts of the other enzymes when pancreatic juice from PM pigs was activated. As expected, the activation of pancreatic juice from CM pigs resulted in the measurement of very high amounts of all the proteolytic enzymes. The amounts of anodal trypsin, chymotrypsins A and B, and elastase II were higher in activated pancreatic juice from CM pigs than from PM pigs.

Collection of pancreatic juice from growing pigs. A comparative study of the pouch method and the catheter method.

CONCLUSION: The results of this study demonstrate that there are large differences in the amount of pancreatic juice secreted and in the chemical and enzymatic composition of pancreatic juice when the pouch and the catheter methods were used, and these differences must be taken into consideration in future studies with either method. METHODS: A study was performed to compare the two most commonly used methods to collect pancreatic juice from growing pigs; namely, the pouch method (PM) and the catheter method (CM). In the first part of the study, three barrows (initial weight 37 kg) were fitted with a pancreatic pouch re-entrant cannula. An isolated pouch was prepared in which the pancreatic duct enters the duodenum. In the second part of the study, also with three barrows (initial weight 32 kg), a catheter was inserted into the pancreatic duct. RESULTS: At several points during the 24-h collection, the hourly rate of pancreatic juice secretion in CM pigs was larger (p < 0.05) than for PM pigs. CM pigs also had a higher (p < 0.05) daily volume of secretion, 4.09 vs 2.63 L/24 h for PM pigs. The pH of pancreatic juice collected from CM pigs was consistently higher (p < 0.01) throughout the 24-h collection. In contrast, the concentration and daily output of bicarbonate did not differ between CM and PM pigs. The concentration of protein in pancreatic juice from PM pigs (7.21 g/L) was higher (p < 0.001) than for CM pigs (4.08 g/L). Specific amylase and lipase and total amylase activities were greater (p < 0.01) in pancreatic juice collected from PM pigs. Specific and total carboxyl ester hydrolase and colipase activities were substantially (p < 0.01) larger in pancreatic juice collected from CM pigs. A major difference between the methods was that trypsin and chymotrypsin were fully active in pancreatic juice from PM pigs, whereas virtually no trypsin or chymotrypsin activity was detected in pancreatic juice from CM pigs. Specific and total chymotrypsin activities did not differ between PM and CM pigs. Both specific and total trypsin activities were substantially higher in pancreatic juice from CM pigs: 3682 U/L and 12,752 U/24 h, respectively, vs 1031 U/L and 2639 U/24 h, respectively, in pancreatic juice from PM pigs.

Exocrine pancreatic secretions in growing pigs fed diets containing fish oil, rapeseed oil or coconut oil.

Two experiments were performed to study the effect of feeding diets containing oils with different fatty acid composition on exocrine pancreatic secretions in growing pigs using two different methods to collect pancreatic juice. In the first experiment, three barrows (initial weight 37 kg) were fitted with a pancreatic pouch re-entrant cannula. An isolated pouch was prepared where the pancreatic duct enters the duodenum. In the second experiment, also using three barrows (initial weight 32 kg), a catheter was inserted into the pancreatic duct. Three wheat starch and fish meal-based diets were formulated to contain either 15 g fish oil, rapeseed oil or coconut oil/100 g. In both experiments, the diets were fed according to a 3 times 3 Latin square design. The volume of pancreatic juice secreted, pH and secretion of bicarbonate, protein, amylase, trypsin, lipase and colipase were not significantly affected by the diets in the first experiment. In the second experiment, chymotrypsin secretion was significantly greater in pigs fed the coconut oil diet, and secretion of carboxyl ester hydrolase was significantly higher in pigs fed the fish oil diet. When compared qualitatively, pigs in Experiment 2 secreted more pancreatic juice; the pancreatic juice had a higher pH, and trypsin, carboxyl ester hydrolase and colipase secretions were substantially higher whereas amylase secretion was lower than for pigs in Experiment 1. The fatty acid composition of the different oils had minor effects on exocrine pancreatic secretion in growing pigs. However, there were considerable differences between the two surgical methods used to collect pancreatic juice, and these differences may be explained by physiological changes induced by the two methods.

Energy metabolism and protein balance in growing rats housed in 18 degree C or 28 degree C environments and fed different levels of dietary protein.

A study was performed to investigate the effect of environmental temperature and increasing levels of protein in the diet on visceral organ size, digestibility, protein balance and energy metabolism in rats. Thirty-six male Wistar rats, initial body weight 77-80 g, were used in a factorial design consisting of three levels of dietary protein and two environmental temperatures of either 18 or 28 degrees C. Three fish meal-based diets were prepared to contain 91, 171 and 262 g protein (N X 6.25/kg diet). Gas-exchange measurements were made and urine and feces were quantitatively collected. The weights of the visceral organs from rats housed at 18 degrees C were greater (P < 0.05) than those of rats housed at 28 degrees C. The digestibilities of dry matter and protein were not affected by environmental temperature, whereas fat and energy digestibilities were higher (P < 0.05) at 18 degrees C than at 28 degrees C. As the level of protein was increased, the digestibilities of protein, energy and fat increased (P < 0.05). Protein intake and protein retention were higher at 18 degrees C (P < 0.05) than at 28 degrees C and increased (P < 0.05) as dietary protein concentration increased. Apparent biological value was lower (P < 0.05) at 18 degrees C than at 28 degrees C and decreased (P < 0.05) as dietary protein level increased. Heat production as a percentage of metabolizable energy was higher (P < 0.05) for the low protein diet than for the medium and high protein diets. The efficiency of energy utilization was depressed (P < 0.05) for the high protein diet when rats were kept at 28 degrees C. The results suggest that thermogenesis was induced when low protein was fed. The increase in thermogenesis may have been important in regulating energy balance and maintaining a constant body temperature in a cold environment.

Effect of beta-glucanase supplementation to hulless barley- or wheat-soybean meal diets on the digestibilities of energy, protein, beta-glucans, and amino acids in young pigs.

Studies were carried out to determine the effect of beta-glucanase supplementation to hulless barley-soybean meal (HB+SBM) or wheat-soybean meal (W+SBM) diets on the digestibilities of GE, CP, beta-glucans, and amino acids. Twelve barrows, average BW 7.3 kg, were fitted with a simple T-cannula at the distal ileum, approximately 5 cm from the ileo-cecal sphincter. After a 7-d recuperation period, six pigs were allotted to each dietary treatment according to a two-period crossover design. Both diets were formulated to contain 20% CP without and with supplementation of .2% beta-glucanase. beta-glucanase refers to a mixture of enzymes with endo- and exo-beta-glucanase and beta-glucosidase activities. Chromic oxide was included as a digestibility marker. The pigs were fed three times daily at 0800, 1600, and 2400 and the daily allowance was offered at a rate of 5% of BW. Each experimental period lasted 9 d. Feces were collected for 48 h on d 6 and 7 and ileal digesta for a total of 24 h on d 8 and 9. beta-glucanase supplementation to the HB+SBM diet increased (P < .05 or P < .01) the ileal digestibilities of GE, CP, beta-glucans, and the majority of the amino acids and the fecal digestibilities of GE, CP, and all amino acids measured; the fecal digestibility of beta-glucans in the HB+SBM diet was not affected by beta-glucanase supplementation. There was no effect (P > .05) of beta-glucanase supplementation to the W+SBM diet on the ileal digestibilities of any criteria measured except for beta-glucans (P < .05). The supplementation of beta-glucanase to the W+SBM diet increased (P < .05) the fecal digestibility of energy but not (P > .05) the other criteria measured.