R. T. Zijlstra

Comparison of Ileum Microflora of Pigs Fed Corn-, Wheat-, or Barley-Based Diets by Chaperonin-60 Sequencing and Quantitative PCR

ABSTRACT We have combined the culture-independent methods of high-throughput sequencing of chaperonin-60 PCR product libraries and quantitative PCR to profile and quantify the small-intestinal microflora of pigs fed diets based on corn, wheat, or barley. A total of 2,751 chaperonin-60 PCR product clones produced from samples of ileum digesta were examined. The majority (81%) of these clones contained sequences independently recovered from all three libraries; 372 different nucleotide sequences were identified, but only 14% of the 372 different sequences were recovered from all three libraries. Taxonomic assignments of the library sequences were made by comparison to a reference database of chaperonin-60 sequences combined with phylogenetic analysis. The taxa identified are consistent with previous reports of pig ileum microflora. Frequencies of each sequence in each library were calculated to identify taxa that varied in frequency between the corn, barley, and wheat libraries. The chaperonin-60 sequence inventory was used as a basis for designing PCR primer sets for taxon-specific quantitative PCR. Results of quantitative PCR analysis of ileum digesta confirmed the relative abundances of targeted taxa identified with the library sequencing approach. The results of this study indicate that chaperonin-60 clone libraries can be valid profiles of complex microbial communities and can be used as the basis for producing quantitative PCR assays to measure the abundance of taxa of interest during experimentally induced or natural changes in a community.

Nonstarch Polysaccharides Modulate Bacterial Microbiota, Pathways for Butyrate Production, and Abundance of Pathogenic Escherichia coli in the Pig Gastrointestinal Tract†

ABSTRACT The impact of nonstarch polysaccharides (NSP) differing in their functional properties on intestinal bacterial community composition, prevalence of butyrate production pathway genes, and occurrence of Escherichia coli virulence factors was studied for eight ileum-cannulated growing pigs by use of terminal restriction fragment length polymorphism (TRFLP) and quantitative PCR. A cornstarch- and casein-based diet was supplemented with low-viscosity, low-fermentability cellulose (CEL), with high-viscosity, low-fermentability carboxymethylcellulose (CMC), with low-viscosity, high-fermentability oat β-glucan (LG), and with high-viscosity, high-fermentability oat β-glucan (HG). Only minor effects of NSP fractions on the ileal bacterial community were observed, but NSP clearly changed the digestion in the small intestine. Compared to what was observed for CMC, more fermentable substrate was transferred into the large intestine with CEL, LG, and HG, resulting in higher levels of postileal dry-matter disappearance. Linear discriminant analysis of NSP and TRFLP profiles and 16S rRNA gene copy numbers for major bacterial groups revealed that CMC resulted in a distinctive bacterial community in comparison to the other NSP, which was characterized by higher gene copy numbers for total bacteria, Bacteroides-Prevotella-Porphyromonas, Clostridium cluster XIVa, and Enterobacteriaceae and increased prevalences of E. coli virulence factors in feces. The numbers of butyryl-coenzyme A (CoA) CoA transferase gene copies were higher than those of butyrate kinase gene copies in feces, and these quantities were affected by NSP. The present results suggest that the NSP fractions clearly and distinctly affected the taxonomic composition and metabolic features of the fecal microbiota. However, the effects were more linked to the individual NSP and to their effect on nutrient flow into the large intestine than to their shared functional properties.

Voluntary feed intake in growing-finishing pigs: A review of the main determining factors and potential approaches for accurate predictions

The ability of pigs to consume sufficient nutrients for optimal performance is an important consideration in commercial pork production. Nutrient intake levels are directly related to voluntary feed intake. Voluntary feed intake in pigs is influenced by several factors including environmental conditions (e.g. thermal and social conditions), animal status (e.g., age and physiological status), and feed and feeding conditions (e.g. bulkiness of the feed and feed form). Although the individual effects of many of these factors on voluntary feed intake have been investigated and quantified, little has been done to characterize their interactive effects. Under commercial conditions, voluntary feed intake is clearly influenced by multiple factors at any one time. Thus, there is a need for a means to accurately quantify voluntary feed intake in pigs as affected by the different interacting factors. Until quantitative effects of these interactions are established it is suggested that feed intake be monitored. This ...

Nonruminant Nutrition Symposium: Controlling feed cost by including alternative ingredients into pig diets: a review.

Sustained price increases for traditional cereal grain and protein meal feed commodities have forced the pork industry to consider the dietary inclusion of alternative feedstuffs. Crop seed may serve as feedstuffs but their demand as feedstock for human food, biofuel, and bioindustrial products has increased. Together with these products, coproducts such as distillers dried grains with solubles, wheat millrun, and canola meal are produced. As omnivores, pigs are ideally suited to convert these non-human-edible coproducts into high-quality food animal protein. Therefore, coproducts and other low-cost alternative feedstuffs such as pulses and oilseeds can be included in pig diets to reduce feed cost per metric ton of feed. However, inclusion of alternative feedstuffs in pig diets does not necessarily reduce feed cost per kilogram of gain. Therefore, the use of novel and existing feedstuffs in pig diets must be optimized following their characterization for energy and AA profile. Alternative feedstuffs generally have a high content of at least 1 of the following antinutritional factors (ANF): fiber, tannins, glucosinolates, and heat-labile trypsin inhibitors. Several methods can optimize nutrient use of pigs fed alternative feedstuffs by reducing effects of their ANF. These methods include 1) particle size reduction to increase nutrient digestibility, 2) dehulling or scarification to reduce tannin and fiber content of pulses and oilseeds, 3) air classification to create fractions that have a greater content of nutrients and lower content of ANF than the feedstock, 4) heat treatments such as extrusion, toasting, roasting, and micronization to reduce heat-labile ANF, 5) dietary supplementation with fiber-degrading enzymes or predigestion of fibrous feedstuffs or diets with fiber-degrading enzymes to increase dietary nutrient availability, and 6) formulation of diets based on bioavailable AA coefficients. In conclusion, the feeding of alternative ingredients may reduce feed cost per unit of pork produced provided that their price per unit NE or digestible lysine is less than that of the traditional feedstuffs and that negative effects of their ANF are controlled.

Effects of xylanase supplementation on the apparent digestibility and digestible content of energy, amino acids, phosphorus, and calcium in wheat and wheat by-products from dry milling fed to grower pigs.

Wheat by-products are feedstuffs that vary in nutritional value, partly because of arabinoxylans that limit nutrient digestibility. Millrun is a byproduct from dry milling wheat into flour and contains varying amounts of the bran, middlings, screening, and shorts fractions. The digestible nutrient content of mill-run is not well known. Effects of xylanase supplementation (0 or 4,000 units/kg of diet) on energy, AA, P, and Ca digestibilities were studied in a wheat control diet and 5 diets containing 30% of a by-product (mill-run, middlings, shorts, screening, or bran) in a 2 x 6 factorial arrangement of treatments. The wheat control diet was formulated to contain 3.34 Mcal of DE/kg and 3.0 g of standardized ileal digestible Lys/Mcal of DE. Diets contained 0.4% chromic oxide. Each of 12 ileal-cannulated pigs (32.5 +/- 2.5 kg) was fed 6 or 7 of 12 diets at 3 times the DE requirement for maintenance in successive 10-d periods for 6 or 7 observations per diet. Feces and ileal digesta were each collected for 2 d. Xylanase tended to increase (P < 0.10) ileal energy digestibility by 2.2 percentage units and the DE content by 0.10 Mcal/kg of DM and increased (P < 0.05) ileal DM digestibility by 2.8 percentage units; a diet x xylanase interaction was not observed. Xylanase increased (P < 0.05) total tract energy and DM digestibilities and the DE content. A diet x xylanase interaction was observed; xylanase increased (P < 0.05) total tract energy digestibility of the millrun diet from 72.1 to 78.9%, DE content from 3.19 to 3.51 Mcal/kg of DM, and DM digestibility from 71.5 to 78.6%. Diet affected (P < 0.05) and xylanase improved (P < 0.05) digestibility and digestible contents of some AA in diets and by-products, including Lys, Thr, and Val. Xylanase increased (P < 0.05) Lys digestibility by 13.8, 5.0, 5.2, 6.0, and 14.1 percentage units in millrun, middlings, shorts, screening, and bran, respectively. Diet affected (P < 0.01) total tract P and Ca digestibilities. Xylanase increased (P < 0.05) digestible P and Ca contents. In summary, nutrient digestibility varies among wheat by-products. Millrun contained 2.65 Mcal of DE/kg of DM, which xylanase increased to 3.56 Mcal of DE/kg of DM. Xylanase improved nutrient digestibility and DE content in wheat by-products; and the extent of improvement depended on the by-product. Xylanase supplementation may maximize opportunities to include wheat byproducts in swine diets and ameliorate reductions in nutrient digestibility that may be associated with arabinoxylans.

Starch with High Amylose Content and Low In Vitro Digestibility Increases Intestinal Nutrient Flow and Microbial Fermentation and Selectively Promotes Bifidobacteria in Pigs

Diets containing different starch types can affect enzymatic digestion of starch and thereby starch availability for microbial fermentation in the gut. However, the role of starch chemistry in nutrient digestion and flow and microbial profile has been poorly explained. Eight ileal-cannulated pigs (29.4 ± 0.9 kg body weight) were fed 4 diets containing 70% purified starch (amylose content, <5, 20, 28, and 63%; reflected by in vitro maximal digestion rate; 1.06, 0.73, 0.38, and 0.22%/min, respectively) in a replicated 4 × 4 Latin square. Ileal and fecal starch output, postileal crude protein yield, fecal total SCFA and total butyrate content, and gene copies of Bifidobacterium spp. in feces were higher (P < 0.05) when pigs consumed the slowly digestible starch diet than the remaining 3 starch diets. The in vitro starch digestion rate had a negative, nonlinear relationship with ileal starch flow (R(2) = 0.98; P < 0.001). Ileal starch flow was positively related to Bifidobacterium spp. (R(2) = 0.27; P < 0.01), Lactobacillus group (R(2) = 0.22; P < 0.01), and total butyrate content (R(2) = 0.46; P < 0.01) but was not related to Enterobacteriaceae (R(2) < 0.00; P = 0.92). In conclusion, starch with high amylose content and low in vitro digestibility increased postileal nutrient flow and microbial fermentation and selectively promoted Bifidobacterium spp. in the distal gut.

The nutritional value of expeller-pressed canola meal for grower-finisher pigs.

Expeller-pressed (EP) canola meal contains more residual oil than solvent-extracted canola meal and might be an attractive feedstuff for swine, but it has been poorly characterized. In Exp. 1, six ileal-cannulated barrows (36 kg of BW) were fed at 3x maintenance either a 44% EP canola meal diet or a N-free diet in a crossover design to measure energy and AA digestibility and calculate standardized ileal digestible (SID) AA and NE content, with 6 observations per diet. Each period consisted of a 5-d diet adaptation and a 2-d feces and 3-d digesta collection. The EP canola meal contained (% of DM) 38.5% CP, 13.3% ether extract, 2.42% Lys, 1.54% Thr, 0.62% Met, and 23.2 micromol/g of glucosinolates. Apparent total tract energy digestibility was 75.0% and the DE and predicted NE content were 3.77 and 2.55 Mcal/kg (in DM), respectively. The SID AA content (% of DM) was 1.77% Lys, 1.04% Thr, and 0.52% Met. In Exp. 2, a total of 1,100 pigs (25 kg of BW) housed in 50 pens were fed 5 dietary regimens with 0, 7.5, 15, and 22.5% or decreasing amounts (22.5, 15, 7.5, and 0%, respectively) of EP canola meal over 4 phases to validate performance and carcass characteristics. Diets were formulated to contain equal NE:SID Lys for each growth phase (g/Mcal; 4.04, d 0 to 25; 3.63, d 26 to 50; 3.23, d 51 to 77; 2.83, d 78 to 90). At slaughter, carcass characteristics were measured for all pigs, and jowl fat was sampled for 2 pigs per pen. For d 51 to 90, the 22.5% EP canola meal regimen was reduced to 18% (22.5/18%) because of decreased ADFI in phases 1 and 2. Overall (d 0 to 90), increasing dietary EP canola meal linearly decreased (P < 0.001) ADG and ADFI and linearly increased (P < 0.01) G:F. For 0 and 22.5/18% EP canola meal, respectively, ADG was 978 and 931 g/d, ADFI was 2.77 and 2.58 kg/d, and G:F was 0.366 and 0.378. Increasing dietary EP canola meal did not alter the carcass backfat thickness, loin depth, or jowl fat fatty acid profile. Pigs fed 22.5/18% EP canola meal reached slaughter weight 3 d after (P < 0.05) pigs fed 0% EP canola meal. In summary, EP canola meal provided adequate energy and AA; however, ADG was reduced by 3 g/d per 1% of EP canola meal inclusion, likely because of increased dietary glucosinolates. Thus, the amount of EP canola meal included in swine diets should be targeted to an expected growth performance and carcass quality. Finally, diets formulated to contain an equal NE and SID AA content did not entirely eliminate the risks for reduced growth performance associated with inclusion of an alternative feedstuff.

Orally administered iodinated recombinant human insulin-like growth factor-I (125I-rhIGF-I) is poorly absorbed by the newborn piglet

BACKGROUND The purpose of the current study was to determine the degree to which milk-borne insulin-like growth factor-I (IGF-I) is absorbed. METHODS Cesarean-derived piglets were fitted with umbilical arterial and venous catheters within 2 h of birth and were administered formula containing 21.7 +/- 1.8 microCi of iodinated recombinant human IGF-1 (125I-rhIGF-I) by orgogastric gavage. Blood samples were taken before administration of the 125I-rhIGF-I (t0) and for 4 h postgavage. Plasma was obtained by centrifugation and total and trichloroacetic acid precipitable radioactivity were determined. Immunoreactive 125I-rhIGF-I was assessed using a polyclonal antibody to human IGF-I. Four hours after feeding, intestines were removed, divided into 13 segments, and flushed with saline. Radioactivity within the small intestinal lumen and wall were measured. RESULTS Radioactivity in portal blood was higher than t0 at all times points (p < 0.05), whereas arterial radioactivity did not differ from t0 until 30 min postgavage. On average 18-20% of total radioactivity in both portal and arterial blood was acid-precipitable, with the proportion decreasing over time (p < 0.001). Immunoprecipitable radioactivity averaged 3-5% of the total radioactivity and was higher in portal than arterial blood (p < 0.05). Based on a plasma volume of 0.062 +/- 0.005 L and a baseline plasma IGF-I concentration of 1.81 +/- 0.56 nmol/L, absorbed 125I-rhIGF-I represented 0.205% of the total plasma IFG-I pool, whereas 14% of the dose was associated with the lining of the intestine. CONCLUSIONS Absorption of orally administered IGF-I does not contribute significantly to circulating IGF-I.

Starch with High Amylose and Low in Vitro Digestibility Increases Short-Chain Fatty Acid Absorption, Reduces Peak Insulin Secretion, and Modulates Incretin Secretion in Pigs

Diets containing different starch types affect peripheral glucose and insulin responses. However, the role of starch chemistry in kinetics of nutrient absorption and insulin and incretin secretion is poorly understood. Four portal vein-catheterized pigs (35.0 ± 0.2 kg body weight) consumed 4 diets containing 70% purified starch [0-63.2% amylose content and 0.22 (slowly) to 1.06%/min (rapidly) maximum rate of in vitro digestion] for 7-d periods in a 4 × 4 Latin square. On d 7, blood was collected for 12 h postprandial with simultaneous blood flow measurement for determining the net portal appearance (NPA) of nutrients and hormones. The NPA of glucose, insulin, C-peptide, and glucose-dependent insulinotropic polypeptide (GIP) during 0-4 h postprandial were lower (P < 0.05) and those of butyrate and total SCFA were higher (P < 0.05) when pigs consumed the diet containing slowly digestible compared with rapidly digestible starch. The peak NPA of insulin occurred prior to that of glucose when pigs consumed diets containing rapidly digestible starch. The kinetics of insulin secretion had a linear positive relation with kinetics of NPA of glucose (R(2) = 0.50; P < 0.01). In conclusion, starch with high amylose and low in vitro digestibility decreases the kinetics of glucose absorption and insulin and GIP secretion and increases SCFA absorption and glucagon-like peptide-1 secretion. In conclusion, starch with high amylose content and a lower rate and extent of in vitro digestion decreased glucose absorption and insulin secretion and increased SCFA absorption.

Effect of phytase and xylanase supplementation or particle size on nutrient digestibility of diets containing distillers dried grains with solubles cofermented from wheat and corn in ileal-cannulated grower pigs.

Nutrient digestibility in distillers dried grains with solubles (DDGS) is limited by physical constraints such as particle size and by biochemical limitations such as phytate and fiber or nonstarch polysaccharides (NSP). To determine the separate effects of these limitations on nutrient digestibility, ground DDGS (383 µm) supplemented with phytase (0 or 250 units/kg of feed) and xylanase (0 or 4,000 units/kg of feed) was evaluated in a 2 × 2 factorial arrangement of treatments together with unground DDGS (517 µm) and an N-free diet in a 6 × 6 Latin square. Cofermented wheat and corn DDGS contained 8.6% moisture, 31.0% CP, 1.04% Lys, 8.0% ether extract, 2.0% starch, 40% NDF, and 0.85% P (as-is basis). Diets contained 43.7% DDGS as the sole source of AA; the digesta from pigs fed the N-free diet served to subtract basal endogenous AA losses and as control for energy digestibility. Six ileal-cannulated barrows (37.1 ± 0.8 kg of BW) were fed 6 diets at 2.8 × maintenance for DE in six 9-d periods. Feces and ileal digesta were collected for 2 d each. The apparent ileal digestibility (AID) of GE and apparent total tract digestibility (ATTD) of GE and NDF were 2.3, 0.5, and 5.1%-units greater (P < 0.05) for the ground than unground DDGS diet, respectively. Consequently, the ATTD of GE was 1.3%-units greater (P < 0.05) and the DE content was 0.06 Mcal/kg greater (P < 0.05) for ground than unground DDGS, respectively. Grinding of DDGS did not affect (P > 0.05) the ATTD of crude fiber, ADF, P, and Ca in diets. Grinding of DDGS increased (P < 0.05) the AID of most AA in diets including Lys, Met, and Thr by 6.9, 1.1, and 1.7%-units, respectively. Grinding of DDGS increased (P < 0.05) the SID of Lys by 6.2%-units and SID content of Lys and Thr by 0.06 and 0.02%-units, respectively. Phytase and xylanase did not interact (P > 0.05) to affect nutrient digestibility. Phytase increased (P < 0.001) the ATTD of P by 10.5%-units, but did not affect (P > 0.05) AA digestibility. Xylanase did not affect nutrient digestibility. In conclusion, particle size is an important physical characteristic affecting digestibility of energy and AA, but not P in DDGS. Phytate in DDGS limits digestibility of P, but not energy and AA. The substrate for xylanase in DDGS did not hinder energy and AA digestibility.