Prajwal R. Regmi

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.

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.

Prediction of in vivo apparent total tract energy digestibility of barley in grower pigs using an in vitro digestibility technique

The DE content within cereal grains can vary 25% mainly due to changes in apparent total tract digestibility (ATTD) of energy. In vitro digestibility techniques have been developed to predict the DE value among feedstuffs. However, these techniques have not been tested properly for their suitability to predict the variation in energy digestibility and DE content within a cereal grain. The objective of the present study was to establish and evaluate an in vitro digestibility technique to predict in vivo ATTD of energy of barley in grower pigs. Barley grain samples (hulled, n = 21) with a large range in quality were collected; the ADF and CP content ranged from 4.5 to 11.4% and 10.0 to 16.4% (DM basis), respectively. The ATTD of energy was determined using barrows (n = 63, 33 +/- 2.1 kg of initial BW) in 2 periods with 6 observations per sample and ranged from 51.9 to 78.5%, with relative errors between 0.4 and 5.0%. A preliminary study, comparing a 2- and a 3-step in vitro digestibility technique using 3 barley samples, indicated that R(2) between in vivo and in vitro energy digestibility was greater using the 3- than the 2-step technique (0.92 vs. 0.76). Therefore, the 3-step in vitro digestibility technique was used solely in subsequent analyses. Briefly, ground barley was subsequently incubated with pepsin for 6 h, pancreatin for 18 h, and cellulase for 24 h. The DM and GE content of samples and residues were measured to calculate digestibility. The in vitro energy digestibility of the 21 barley samples with duplicate measurements ranged from 63.7 to 82.2%, with relative errors between 0.1 and 2.6%. In vitro energy digestibility was strongly related (y = 1.25 x - 25.22; R(2) = 0.81) to in vivo energy digestibility. Finally, a subset of 7 barley samples was analyzed in quadruplicate using the 3-step in vitro technique. The relationship between in vitro and in vivo energy digestibility was very strong (y = 1.23 x - 25.33; R(2) = 0.97) with relative errors between 0.5 and 2.7%. In vitro DE and energy digestibility were perfectly related (R(2) = 1.00). In summary, the 3-step in vitro energy digestibility technique can accurately predict the ATTD of energy in barley in grower pigs. The 3-step in vitro digestibility technique, thus, might be useful as the reference laboratory procedure to calibrate analytical equipment to rapidly predict the ATTD of energy in barley.

In vitro digestibility techniques to predict apparent total tract energy digestibility of wheat in grower pigs

In vitro digestibility techniques have been developed to predict the apparent total tract digestibility (ATTD) of energy and DE content of mixed diets and feedstuffs including barley grain in swine. However, the techniques have not been tested properly for their accuracy in predicting the variation in ATTD of energy and DE content within wheat grain. The objectives were 1) to compare two 3-step in vitro digestibility techniques with either cellulase (IVD-CEL) or Viscozyme (a multienzyme complex to digest fiber; Novozymes, Bagsvaerd, Denmark; IVD-VIS) as the third step, and differing in the amount of enzymes used and the duration of digestion, for their accuracy in predicting ATTD of energy and DM of wheat in grower pigs; and 2) to develop equations to predict ATTD of energy of different batches of wheat. Wheat grain samples (n = 20) with a wide range in quality were collected; the ADF and CP content ranged from 3.3 to 6.2% and from 11.2 to 20.8% (DM basis), respectively. The ATTD of energy was determined using barrows (n = 60, 30.7 +/- 4.7 kg of initial BW) in 2 periods with 6 observations per sample, and ranged from 73.3 to 84.5%. In IVD-CEL, 1 g of ground wheat was digested sequentially in digestion solutions containing pepsin (10 mg/36.5 mL) for 6 h, pancreatin (150 mg/54.5 mL) for 18 h, and cellulase (75 mg/55.5 mL) for 24 h. In IVD-VIS, 0.5 g of ground wheat sample was digested sequentially in solutions containing pepsin (25 mg/36.5 mL) for 2 h, 3 mL of pancreatin (100 mg/54.5 mL) for 6 h, and Viscozyme (0.5 mL/65.3 mL) for 18 h. The in vitro energy and DM digestibility ranged from 79.8 to 91.0% and from 82.0 to 91.5% for IVD-CEL, and ranged from 76.2 to 87.0% and from 79.1 to 89.4% for IVD-VIS, respectively. The R(2) between ATTD of energy and in vitro DM and energy digestibility for IVD-VIS (0.82 and 0.73, respectively) was greater than for IVD-CEL (0.55 and 0.54, respectively). The equation y = 1.05x - 8.85 using the in vitro DM digestibility value from IVD-VIS can predict the ATTD of the energy of wheat in swine with an SE of prediction of 1.2. The relationship between in vitro DM digestibility and grain characteristics such as ADF was stronger for the IVD-VIS than for the IVD-CEL technique (R(2) = 0.89 vs. 0.70). In conclusion, the IVD-VIS, but not the IVD-CEL, technique can accurately (R(2) = 0.82) predict the ATTD of energy in wheat in grower pigs. Therefore, the IVD-VIS technique might be useful as the reference analysis to calibrate analytical equipment to predict the ATTD of energy rapidly in wheat.

High Amylose Starch with Low In Vitro Digestibility Stimulates Hindgut Fermentation and Has a Bifidogenic Effect in Weaned Pigs

BACKGROUND Dietary amylose resists enzymatic digestion, thereby providing a substrate for microbial fermentation that stimulates proliferation of beneficial microbiota and production of short-chain fatty acids (SCFAs) in the large intestine of pigs and humans. However, the effect of increasing dietary amylose in pigs immediately postweaning on growth, nutrient digestibility and flow, and intestinal microbial and SCFA profiles has not been studied and can be used as a model for newly weaned human infants. OBJECTIVE We studied the effects of increasing dietary amylose on growth, nutrient digestibility, and intestinal microbial and metabolite profiles in weaned pigs. METHODS Weaned pigs (n = 32) were randomly allocated to 1 of 4 diets containing 67% starch with 0%, 20%, 28%, or 63% amylose for 21 d. Subsequently, pigs were killed to collect feces and digesta for measuring starch digestion and microbial and metabolite profiles. RESULTS Feeding weaned pigs 63% compared with 0%, 20%, and 28% amylose decreased (P < 0.05) feed intake by 5% and growth by ≥ 12%. Ileal digestibility of dry matter decreased (P < 0.05) by 10% and starch by 9%, thereby increasing (P < 0.05) hindgut fermentation, cecal and colonic total SCFAs, and colonic Bacteroides, and lowering (P < 0.01) ileal, cecal, and colonic pH in pigs consuming 63% compared with 0%, 20%, and 28% amylose. Cecal and colonic Bifidobacteria spp. increased by 14-30% (P < 0.05) and Clostridium clusters IV and XIVa were decreased (P < 0.01) in pigs consuming 63% compared with 0%, 20%, and 28% amylose. CONCLUSION Increasing dietary amylose in pigs immediately postweaning stimulated hindgut fermentation and Bifidobacteria spp., thereby manipulating the gut environment, but also reduced intake and growth. An optimum dietary amylose concentration should be determined, which would maintain desired growth rate and gut environment in weaned pigs.

Slowly digestible starch influences mRNA abundance of glucose and short-chain fatty acid transporters in the porcine distal intestinal tract.

The relationship between starch chemistry and intestinal nutrient transporters is not well characterized. We hypothesized that inclusion of slowly instead of rapidly digestible starch in pig diets will decrease glucose and increase short-chain fatty acid (SCFA) transporter expression in the distal gut. Weaned barrows (n = 32) were fed 4 diets containing 70% starch [ranging from 0 to 63% amylose and from 1.06 (rapidly) to 0.22%/min (slowly) rate of in vitro digestion] at 3 × maintenance energy requirement in a complete randomized block design. Ileal and colon mucosa was collected on day 21 to quantify mRNA abundance of Na(+)-dependent glucose transporter 1 (SGLT1), monocarboxylic acid transporter 1 (MCT1), and Na(+)-coupled monocarboxylate transporter (SMCT). Messenger RNA was extracted and cDNA manufactured prior to relative quantitative reverse transcription PCR. Data were analyzed using the 2(-Δ ΔC)(T) method, with β-actin and glyceraldehyde-3-phosphate dehydrogenase as reference genes, and regression analysis was performed. As in vitro rate of digestion decreased, SGLT1 linearly increased (P < 0.05) in the ileum. Contrary to SGLT1, MCT1 tended to linearly decrease (P = 0.08) in the ileum and increased quadratically (P < 0.001) in the colon with decreasing rate of digestion. Starch digestion rate did not affect SMCT in the ileum; however, colonic SMCT quadratically decreased (P < 0.01) with decreasing rate of digestion. In conclusion, in contrast to our hypothesis, slowly digestible starch increased ileal glucose and decreased ileal SCFA transporter mRNA abundance, possibly due to an increased glucose in the luminal ileum. Effects of starch on colonic SCFA transporter mRNA abundance were inconsistent.

Effects of ammonia load on glucose metabolism by isolated ovine duodenal mucosa

To determine the effects of ammonia load on glucose metabolism in ruminant small intestinal tissues, duodenal mucosal cells (DMC) were isolated from growing female sheep (n = 10; 46. 0 +/- 0. 8 kg of BW) fed diets differing in CP content: high (19. 4%) vs. low (13. 1%). Ammonia concentration in the duodenal digesta fluid was greater for sheep fed a high CP diet compared with those fed a low CP diet (16. 4 +/- 1. 0 vs. 9. 1 +/- 1. 8 mM). The isolated primary mucosal cells were incubated for 90 min with [2-(13)C] glucose (3 mM) and ammonium chloride (0, 0. 1, 1, 5, 10, 20, or 50 mM) in Krebs-Ringer HEPES buffer. It was hypothesized that DMC would increase glucose carbon utilization for the synthesis of nonessential AA when the ammonia concentration in the incubation media increased. However, utilization of glucose carbon for alanine synthesis decreased linearly (P = 0. 03) as the ammonia concentration in the incubation media increased. Furthermore, glucose disappearance and utilization of glucose carbon for aspartate synthesis were not affected (P > 0. 47) by the ammonia concentration. Contrarily, in vitro glucose disappearance was greater (P = 0. 03) for DMC isolated from sheep fed a low CP diet vs. a high CP diet [14. 6 +/- 1. 6 vs. 8. 6 +/- 1. 3 nmol.(10(6) cells)(-1).(90 min) (-1)], and hexokinase activity was greater (P = 0. 01) in the mucosa of sheep fed a low CP diet compared with a high CP diet (1. 22 +/- 0. 05 vs. 1. 04 +/- 0. 02 mUnit/mg of protein). These observations indicate that ammonia load does not affect the extent of glucose utilization by DMC, and that glucose carbon may not play a significant role for the synthesis of alanine, aspartate, or glutamate when DMC are exposed to increased concentrations of ammonia.