Transport of valine across the small intestinal epithelium in pigs fed different valine levels and Bacillus subtilis.

Mutants of Bacillus subtilis overproducing valine (B. subtilis VAL) could be an approach to supply pigs dietary valine (Val). In the study, 18 gilts were fed: (i) negative diet with a standardized ileal digestible (SID) Val:Lys of 0.63:1 (Neg); (ii) Neg added B. subtilis VAL (1.28 × 1011  cfu/kg as-fed) or; (iii) Neg added L-Val to a Val:Lys of 0.69:1. Using the Ussing chamber method, the study aimed to investigate whether (i) the diets affect intestinal transport of additions of 0, 5, 10 or 20 mmol Val/L from the mucosal to the serosal side and (ii) the B. subtilis VAL contributes to a net transport of Val produced in situ. The results showed that the Isc (ΔIscVal ) and release of Val to the serosal side solution (Srel ; µmol cm-2  min-1 ) increased with Val addition (linear and quadratic, p < .0001) but was similar for 5, 10 or 20 mmol Val/L and not affected by diet. No net transport of in situ produced Val by B. subtilis VAL was detected. In conclusion, feeding a Val-deficient diet with or without B. subtilis VAL or a Val sufficient diet did not affect the Val transport across intestinal epithelia. No in situ Val production by B. subtilis VAL was observed in the Ussing chambers.

Evaluation of in situ valine production by Bacillus subtilis in young pigs.

Mutants of Bacillus subtilis can be developed to overproduce Val in vitro. It was hypothesized that addition of Bacillus subtilis mutants to pig diets can be a strategy to supply the animal with Val. The objective was to investigate the effect of Bacillus subtilis mutants on growth performance and blood amino acid (AA) concentrations when fed to piglets. Experiment 1 included 18 pigs (15.0±1.1 kg) fed one of three diets containing either 0.63 or 0.69 standardized ileal digestible (SID) Val : Lys, or 0.63 SID Val : Lys supplemented with a Bacillus subtilis mutant (mutant 1). Blood samples were obtained 0.5 h before feeding and at 1, 2, 3, 4, 5 and 6 h after feeding and analyzed for AAs. In Experiment 2, 80 piglets (9.1±1.1 kg) were fed one of four diets containing 0.63 or 0.67 SID Val : Lys, or 0.63 SID Val : Lys supplemented with another Bacillus subtilis mutant (mutant 2) or its parent wild type. Average daily feed intake, daily weight gain and feed conversion ratio were measured on days 7, 14 and 21. On day 17, blood samples were taken and analyzed for AAs. On days 24 to 26, six pigs from each dietary treatment were fitted with a permanent jugular vein catheter, and blood samples were taken for AA analysis 0.5 h before feeding and at 1, 2, 3, 4, 5 and 6 h after feeding. In experiment 1, Bacillus subtilis mutant 1 tended (P<0.10) to increase the plasma levels of Val at 2 and 3 h post-feeding, but this was not confirmed in Experiment 2. In Experiment 2, Bacillus subtilis mutant 2 and the wild type did not result in a growth performance different from the negative and positive controls. In conclusion, results obtained with the mutant strains of Bacillus subtilis were not better than results obtained with the wild-type strain, and for both strains, the results were not different than the negative control.

Requirement of standardized ileal digestible valine to lysine ratio for 8- to 14-kg pigs.

The objective was to define the Val requirement for weaned piglets in the context of reducing the dietary protein content. A dose-response experiment was conducted to estimate the standardized ileal digestible (SID) Val to Lys ratio required to support the optimum growth of post-weaned piglets. In this study, 96 pigs weighing 8 kg were allotted to one of six dietary treatments (16 pigs for each dietary treatment) and were housed individually. Diets were formulated to provide 0.58, 0.62, 0.66, 0.70, 0.74 and 0.78 SID Val : Lys by adding graded levels of crystalline l-Val to the 0.58 SID Val : Lys diet. Lysine was sub-limiting and supplied 90% of the recommendation (10.95 g SID Lys/kg equal to 11.8 g/kg total Lys). Average daily feed intake (ADFI), average daily gain (ADG) and gain to feed ratio (G : F) were determined during a 14-day period of ad libitum feeding. Blood and urine samples were taken at the end of each week (day 7 and 14 of the experiment) 3 h after feeding the experimental diets. The maximum ADFI and ADG were obtained in pigs fed the 0.78 SID Val : Lys diet; it was not different from the results of pigs fed 0.70 SID Val : Lys diet. The highest G : F was obtained in pigs fed 0.70 SID Val : Lys. The plasma concentration of Val increased linearly (P<0.001) as the dietary SID Val : Lys increased. The increasing dietary Val : Lys also resulted in a linear increase in Cys (P<0.001) and a quadratic increase in Arg (P=0.003), Lys (P=0.05) and Phe (P=0.009). The plasma Gly showed a quadratic decrease (P=0.05) as the dietary Val : Lys increased. Neither plasma nor urinary urea to creatinine ratio was affected by treatment. The minimum SID Val : Lys required to maximize ADFI, ADG and G : F was estimated at 0.67 SID Val : Lys by a broken-line model, and at 0.71 SID Val : Lys by a curvilinear plateau model. The Val deficiency caused a reduction in ADFI, and Val supplementation above the requirement did not impair animal performance. In conclusion, 0.70 SID Val : Lys is suggested as the Val requirement for 8 to 14 kg individually housed pigs.

Microbial phytase addition resulted in a greater increase in phosphorus digestibility in dry-fed compared with liquid-fed non-heat-treated wheat-barley-maize diets for pigs.

The objective was to evaluate the effect of microbial phytase (1250 FTU/kg diet with 88% dry matter (DM)) on apparent total tract digestibility (ATTD) of phosphorus (P) in pigs fed a dry or soaked diet. Twenty-four pigs (65±3 kg) from six litters were used. Pigs were housed in metabolism crates and fed one of four diets for 12 days; 5 days for adaptation and 7 days for total, but separate collection of feces and urine. The basal diet was composed of wheat, barley, maize, soybean meal and no mineral phosphate. Dietary treatments were: basal dry-fed diet (BDD), BDD with microbial phytase (BDD+phy), BDD soaked for 24 h at 20°C before feeding (BDS) and BDS with microbial phytase (BDS+phy). Supplementation of microbial phytase increased ATTD of DM and crude protein (N×6.25) by 2 and 3 percentage units (P<0.0001; P<0.001), respectively. The ATTD of P was affected by the interaction between microbial phytase and soaking (P=0.02). This was due to a greater increase in ATTD of P by soaking of the diet containing solely plant phytase compared with the diet supplemented with microbial phytase: 35%, 65%, 44% and 68% for BDD, BDD+phy, BSD and BSD+phy, respectively. As such, supplementation of microbial phytase increased ATTD of P in the dry-fed diet, but not in the soaked diet. The higher ATTD of P for BDS compared with BDD resulted from the degradation of 54% of the phytate in BDS by wheat and barley phytases during soaking. On the other hand, soaking of BDS+phy did not increase ATTD of P significantly compared with BDD+phy despite that 76% of the phytate in BDS+phy was degraded before feeding. In conclusion, soaking of BDS containing solely plant phytase provided a great potential for increasing ATTD of P. However, this potential was not present when microbial phytase (1250 FTU/kg diet) was supplemented, most likely because soaking of BDS+phy for 24 h at 20°C did not result in a complete degradation of phytate before feeding.

Effect of particle size and microbial phytase on phytate degradation in incubated maize and soybean meal.

The objective of the study was to evaluate the effect of screen size (1, 2 and 3 mm) and microbial phytase (0 and 1000 FTU/kg as-fed) on phytate degradation in maize (100% maize), soybean meal (100% SBM) and maize-SBM (75% maize and 25% SBM) incubated in water for 0, 2, 4, 8 and 24 h at 38°C. Samples were analysed for pH, dry matter and phytate phosphorus (P). Particle size distribution (PSD) and average particle size (APS) of samples were measured by the Laser Diffraction and Bygholm method. PSD differed between the two methods, whereas APS was similar. Decreasing screen size from 3 to 1 mm reduced APS by 48% in maize, 30% in SBM and 26% in maize-SBM. No interaction between screen size and microbial phytase on phytate degradation was observed, but the interaction between microbial phytase and incubation time was significant ( P<0.001). This was because microbial phytase reduced phytate P by 88% in maize, 84% in maize-SBM and 75% in SBM after 2 h of incubation ( P<0.05), whereas the reduction of phytate P was limited (<50%) in the feeds, even after 24 h when no microbial phytase was added. The exponential decay model was fitted to the feeds with microbial phytase to analyse the effect of screen size and feed on microbial phytase efficacy on phytate degradation. The interaction between screen size and feed affected the relative phytate degradation rate ( Rd) of microbial phytase as well as the time to decrease 50% of the phytate P ( t1 =2) ( P<0.001). Thus, changing from 3 to 1 mm screen size increased Rd by 22 and 10%/h and shortened t1 =2 by 0.4 and 0.2 h in maize and maize-SBM, respectively ( P<0.05), but not in SBM. Moreover, the screen size effect was more pronounced in maize and maize-SBM compared with SBM as a higher phytate degradation rate constant (Kd) and Rd, and a shorter t1 =2 was observed in maize compared with SBM in all screen sizes ( P<0.05). However, a higher amount of degraded phytate was achieved in SBM than in maize because of the higher initial phytate P content in SBM. In conclusion, reducing screen size from 3 to 1 mm increased Kd and Rd and decreased t1 =2 in maize and maize-SBM with microbial phytase. The positive effect of grinding on improving microbial phytase efficacy, which was expressed as Kd, Rd and t1/2, was greater in maize than in SBM.

Distinct composition of bovine milk from Jersey and Holstein-Friesian cows with good, poor, or noncoagulation properties as reflected in protein genetic variants and isoforms.

The objective of this study was to examine variation in overall milk, protein, and mineral composition of bovine milk in relation to rennet-induced coagulation, with the aim of elucidating the underlying causes of milk with impaired coagulation abilities. On the basis of an initial screening of 892 milk samples from 42 herds with Danish Jersey and Holstein-Friesian cows, a subset of 102 samples was selected to represent milk with good, poor, or noncoagulating properties (i.e., samples that within each breed represented the most extremes in regard to coagulation properties). Milk with good coagulation characteristics was defined as milk forming a strong coagulum based on oscillatory rheology, as indicated by high values for maximum coagulum strength (G'(max)) and curd firming rate (CFR) and a short rennet coagulation time. Poorly coagulating milk formed a weak coagulum, with a low G'(max) and CFR and a long rennet coagulation time. Noncoagulating milk was defined as milk that failed to form a coagulum, having G'(max) and CFR values of zero at measurements taken within 1h after addition of rennet. For both breeds, a lower content of total protein, total casein (CN) and κ-CN, and lower levels of minerals (Ca, P, Mg) were identified in poorly coagulating and noncoagulating milk in comparison with milk with good coagulation properties. Liquid chromatography/electrospray ionization-mass spectrometry revealed the presence of a great variety of genetic variants of the major milk proteins, namely, α(S1)-CN (variants B and C), α(S2)-CN (A), ß-CN (A(1), A(2), B, I, and F), κ-CN (A, B, and E), α-lactalbumin (B), and ß-lactoglobulin (A, B, and C). In poorly coagulating and noncoagulating milk samples of both breeds, the predominant composite genotype of α(S1)-, ß-, and κ-CN was BB-A(2)A(2)-AA, which confirmed a genetic contribution to impaired milk coagulation. Interestingly, subtle variations in posttranslational modification of CN were observed between the coagulation classes in both breeds. Poorly coagulating and noncoagulating milk contained a lower fraction of the least phosphorylated α(S1)-CN form, α(S1)-CN 8P, relative to total α(S1)-CN, along with a lower fraction of glycosylated κ-CN relative to total κ-CN. Thus, apparent variation was observed in the milk and protein composition, in the genetic makeup of the major milk proteins, and in the posttranslational modification level of CN between milk samples with either good or impaired coagulation ability, whereas the composition of poorly coagulating and noncoagulating milk was similar.

Effect of microbial phytase on phosphorus digestibility in non-heat-treated and heat-treated wheat-barley pig diets.

The objective was to evaluate effects of microbial phytase on apparent total tract digestibility (ATTD) of P in a non-heat-treated and a heat-treated wheat (Triticum aestivum)-barley (Hordeum vulgare) diet fed without inorganic P in a 2 × 3 factorial arrangement. The basal diet was ground and half of the batch was steam pelleted at 81°C and crumbled. Phytase was added at 0, 250, and 500 phytase units (FTU)/kg as-fed (Aspergillus niger). The study comprised 36 pigs from 6 litters. Pigs were housed in metabolism crates and fed 1 of 6 diets for 12 d: 5 d for adaptation and 7 d for total collection of feces. The ATTD of P was highest (P < 0.01) for the non-heat-treated diets and highest (P < 0.01) for the phytase-supplemented diets. Heat treatment reduced plant phytase activity by 25% whereby the ATTD of P decreased (P < 0.01) from 57 to 49%. Microbial phytase increased the ATTD of P to a maximum of 64 and 61% in the non-heat-treated and heat-treated diets corresponding to an increase of 7 and 12%-units. Responses for ATTD of P did not differ between 250 vs. 500 FTU/kg as-fed. In conclusion, processing of feed (meal or pellets) containing plant phytase should be considered to avoid over- or underestimation of effects of microbial phytase.

Absorption and metabolism of benzoic acid in growing pigs.

Dietary benzoic acid (BA) supplementation causes a pronounced reduction in urinary pH but only small changes in blood pH. The present study aimed to investigate the portal absorption profile, hepatic metabolism of BA, and renal excretion of hippuric acid (HA) underlying the relatively small impact of BA on systemic acid-base status. Eight growing pigs (BW = 63 +/- 1 kg at sampling) fitted with permanent indwelling catheters in the abdominal aorta, hepatic portal vein, hepatic vein, and mesenteric vein were allocated to 4 sampling blocks and randomly assigned to control (CON; nonsupplemented diet) or BA supplementation (B; control diet + 1% BA top-dressed). Feed intake was restricted to 3.6% of BW and the ration divided into 3 equally sized meals offered at 8-h intervals. Blood pH (7.465 and 7.486 +/- 0.004) and urinary pH (4.99 and 7.01 +/- 0.09) were less (P = 0.03 and P < 0.01) in B compared with CON. The arterial concentration, net portal flux, and net hepatic uptake of BA increased (P < 0.01) in B compared with CON. The net portal flux of BA increased (P < 0.01) after feeding with B, but remained positive (P < 0.01) at all sampling times (n = 8). Recovery of dietary BA as increased net portal flux and hepatic uptake of BA was 87 +/- 5% and 89 +/- 15%, respectively. The recovery of dietary BA as urinary excretion of BA and HA was 0.08 +/- 0.02% and 85 +/- 7%, respectively. It is concluded that the small impact of BA supplementation on systemic acid-base status was caused by a protracted BA absorption and efficient hepatic extraction and glycine conjugation in combination with efficient renal clearance of HA. Together, these physiological mechanisms prevented major BA and HA accumulation in body fluids.

Interactions between phosphorus feeding strategies for pigs and dairy cows and separation efficiency of slurry.

Phosphorus (P) in manure is a nutrient source for plants, but surplus P amended to fields represents a risk to the environment. This study examines the interactions between low-P diets for pigs and dairy cows and the separation of animal slurry into a solid P fraction and a liquid fraction. Replacing inorganic phosphates with phytase in pig feed reduced the concentration of P in slurry by 35%, but supplementing concentrates to dairy cows did not affect the P concentration in cattle slurry. Particle-size fractions of the slurry were not affected by these dietary changes. The amount of dry matter (DM) in the < 0.025 mm fraction was greater in pig slurry than in cattle slurry, but the relative amounts of P and nitrogen (N) were larger in the > 0.025 mm fraction. Replacing feed phosphate, in the form of mono-calcium phosphate, with phytase in the pig diet reduced the separation index (efficiency) of P from 80% to 60%.

Assessment of zinc and copper status in weaned piglets in relation to dietary zinc and copper supply.

The aim of the study was to determine the effect of weaning and the effect of increasing dietary zinc concentrations on the zinc and copper status of weaned piglets (study 1) and to study the effect of high concentrations of dietary zinc and/or copper on zinc and copper status of weaned piglets (study 2). Study 1 included 54 piglets (six litters of nine piglets). One piglet from every litter was killed 1 day before weaning. The remaining 48 piglets were allocated at weaning (28 days) to four dietary zinc treatments (100, 250, 1000 or 2500 ppm) and subsequently killed 1-2, 5-6 or 14-15 days after weaning. Study 2 included 48 piglets (six litters of eight piglets) allocated to four dietary treatments, consisting of low or high dietary zinc (100 or 2500 ppm) in combination with low or high dietary copper (20 or 175 ppm). All piglets in study 2 were killed 5-7 days after weaning. In both studies, the trace mineral status was assessed by zinc and copper concentrations and alkaline phosphatase (AP) activity in plasma and mucosal tissue. In study 2, lymphocyte metallothionein (MT) mRNA and intestinal mucosa MT mRNA concentrations were included as zinc status markers. The results showed that the zinc status, measured as zinc in plasma and mucosa, was not affected by weaning of the piglets. Plasma copper concentrations decreased during the first 2 weeks after weaning. High dietary copper concentrations did not affect the concentration of copper in plasma, but increased the concentration of copper in mucosa and the concentration of zinc in plasma. The dietary zinc treatments increased the zinc concentration in plasma as well as the zinc and MT mRNA concentration in mucosa. Lymphocyte MT mRNA concentrations did not reflect the differences in dietary zinc supplementation.

Influence of dietary zinc and copper on digestive enzyme activity and intestinal morphology in weaned pigs.

The current study was conducted to investigate the effects of high dietary concentrations of Zn as zinc oxide and Cu as copper sulfate on the activity of digestive enzymes in the pancreas and the intestinal mucosa, intestinal morphology, and mucin histochemistry in pigs after weaning. Thirty-two pigs were weaned at 4 wk of age. The pigs were fed standard weaning diets supplemented with Zn (100 or 2,500 ppm) and Cu (0 or 175 ppm) in a 2 x 2 factorial arrangement of treatments for a 14-d period. In pancreatic tissue, the activity of amylase, carboxypeptidase A, chymotrypsin, trypsin, and lipase increased (P < 0.01) in pigs fed 2,500 ppm of Zn, whereas the activity of carboxypeptidase B and carboxylester hydrolase was unaffected. Copper had no effect on the activity of pancreatic enzymes. In small intestinal contents, the total activity of amylase and carboxypeptidase A was greater in pigs fed 100 ppm of Zn (P < 0.05), whereas feeding 2,500 ppm of Zn increased the chymotrypsin activity (P < 0.001). The remaining enzymes were unaffected by dietary Zn concentration. The villi were longer in the cranial small intestine (P < 0.001) in pigs fed 100 ppm of Zn than in pigs fed 2,500 ppm of Zn, but otherwise there were no clear effects of Zn and Cu supplementation on intestinal morphology. In the cranial small intestine, the activity of maltase (P < 0.001), sucrase (P < 0.001), and lactase was greater in pigs fed 100 ppm of Zn, even though there was a Zn x Cu interaction (P < 0.05) in lactase activity. In the middle and caudal small intestine, no clear differences between dietary treatments were observed. The activity of gamma-glutamyl transpeptidase in the intestinal mucosa was not affected by dietary Zn or Cu. In pigs fed 100 ppm of Zn, the activity of aminopeptidase N was greater in the caudal small intestine, but dietary Zn or Cu had no effect on aminopeptidase N in the cranial and middle small intestine. No effect of dietary Zn or Cu supplementation was found on carbohydrate histochemistry in the caudal small intestine, whereas high dietary Zn increased the area of neutral, acidic, and sulfomucins in the cecum (P < 0.01) and in the colon (P < 0.001). In summary, high dietary Zn increased the activity of several enzymes in the pancreatic tissue and increased the mucin staining area in the large intestine, whereas Cu had no clear effect on these variables. However, no definite answers were found as to how the growth promoting and diarrhea reducing effects of excess dietary Zn are exerted.

Eating and drinking activity of newly weaned piglets: effects of individual characteristics, social mixing, and addition of extra zinc to the feed.

In production systems, piglets usually fast for a period after weaning, thereby increasing the risk of diarrhea and a reduction in growth. The low level of eating may relate to insufficient drinking activity, as solid feed intake must be accompanied by water intake. Mixing of newly weaned piglets is a well-known stressor and a common procedure in pig production. The effect of mixing on the temporal development of eating and drinking activity in newly weaned piglets has not been elucidated. High concentrations of zinc (Zn) in the feed improve the health and performance of piglets after weaning, but the underlying mechanisms are still obscure. One possibility is that Zn affects eating and drinking behavior. The effects of mixing 4 littermates from each of 2 litters and adding zinc oxide (ZnO; 2,500 ppm of Zn) to the feed were studied in a 2 x 2 factorial experiment using 123 piglets weaned at 27 d of age. Individual eating and drinking times during the initial 48 h after weaning were analyzed on 2 levels of aggregation, day and hour. The piglets spent less time eating on the first day after weaning compared with the second day (20 +/- 5 vs. 98 +/- 10 min, respectively; P < 0.001), whereas they spent more time drinking on the first day compared with the second day (13 +/- 1 vs. 9 +/- 0.5 min, respectively; P < 0.001). Eating and drinking times were positively associated (P < 0.001). Females ate for longer than males (61 +/- 8 vs. 44 +/- 7 min/24 h, respectively, P = 0.002), whereas sex did not affect drinking time. Drinking time increased (P = 0.003) and eating time decreased (P = 0.001) with increasing preweaning growth rate and weaning weight. Neither mixing nor addition of ZnO affected the daily eating time. However, nonmixed piglets given 2,500 ppm of Zn as ZnO in the feed spent more time drinking per day (12 +/- 1 min) than did nonmixed piglets offered 100 ppm of Zn as ZnO (10 +/- 1 min; P = 0.002). Mixing also affected the hourly distribution of the drinking activity (P < 0.05). In conclusion, the drinking behavior of newly weaned piglets was more affected by the external factors, mixing and addition of ZnO to the feed, than the eating behavior. As eating and drinking are strongly associated, more focus should be paid to the water intake and the interplay between eating and drinking behavior in future studies aiming to reduce weaning problems.

In vitro binding capacity of zeolite A to calcium, phosphorus and magnesium in rumen fluid as influenced by changes in pH.

An in vitro experiment was designed to mimic the transport of ingested zeolite A in the forestomachs and proximal part of the small intestine so as to evaluate the binding capacity of zeolite A to Ca, P and Mg as influenced by changes in pH. This was done by incubation of rumen fluid solutions with and without zeolite, as well as varying the content of Ca and/or P. The pH was lowered by addition of HCl so as to mimic abomasal conditions, followed by subsequent HCO3- addition to mimic small intestinal pH. Rumen fluid samples were taken at strategic time points in the experiment. All samples were centrifuged and the supernatant analysed for Ca, P and Mg as indicators of the amount of unbound mineral. The addition of zeolite to rumen fluid solutions reduced the amount of supernatant Ca and Mg at rumen pH, whereas the level of P was not reduced. After adding HCl, a large proportion of the zeolite-bound Ca and Mg was released, increasing supernatant Ca and Mg levels; whereas, HCl addition led to a profound drop in supernatant P in zeolite samples, indicating binding of P. A low level of supernatant P was maintained after HCO3- addition. Neutralization by HCO3- led to a zeolite-induced drop in supernatant Ca and Mg. The reduction in supernatant Ca observed in the present study concurs well with the theoretical rationale of prepartum zeolite supplementation in milk fever prevention. Furthermore, the apparent binding of P by the zeolite may also contribute because of the connection between the calcium and phosphorus homeostasis. The zeolite-induced reduction in supernatant Mg indicates that zeolite supplementation should probably be avoided in Mg-deficient herds unless Mg supplementation is initiated.

Excessive dietary selenium to primiparous sows and their offspring. I. Influence on reproduction and growth.

The effect of high dietary selenium (Se) on the reproductive performance, growth and health in pigs was examined. Addition of 0 to 16 mg Se per kg feed to sows and their piglets up to nine weeks of age did not cause any manifest toxic effect. None of the sows died due to the Se treatment. The piglets were all fullborn and showed no macroscopic abnormalities. The treatment did not influence neither the number of liveborn and stillborn piglets in the litter, nor the survival of the piglets until 9 weeks of age. The weight of the whole litter at birth was unaffected by the Se supplementation, while there was a significant difference in body weight of the piglets at 9 weeks of age. The weaned pigs receiving 8 or 16 mg Se per kg feed had a reduced feed intake. As the feed utilization was unaffected by treatment, these pigs had a significantly lower weight at 9 weeks of age.