R. O. Ball

Novel Neonatal Piglet Models of Surgical Short Bowel Syndrome With Intestinal Failure

Objectives: Short bowel syndrome occurring after surgery for acquired or congenital intestinal abnormalities causes considerable neonatal morbidity and mortality. Animal models are a valuable research tool for this problem; however, few successful neonatal models have been developed and most do not include distal intestinal resection as seen commonly in human babies. We report novel piglet models addressing these gaps. Subjects and Methods: Neonatal piglets (1–6 days) underwent venous and gastric catheter insertion and 75% intestinal resection. Group 1 (n = 6) had midintestinal resection with jejunoileal anastomosis; group 2 (n = 5) had distal intestinal resection with jejunocolic anastomosis; group 3 (n = 5) were sham controls; and group 4 (n = 5) were sow reared. Postoperatively, groups 1 to 3 piglets commenced parenteral nutrition (PN), and enteral nutrition was introduced and advanced using a standard regimen. Data collection included days on PN, weight gain, fat absorption, small intestine lengthening, and bowel/liver histology. Results: Group 2 piglets had more days on PN (P = 0.008), less weight gain (P = 0.027), and greater malabsorption (P = 0.012). They did not show small intestine lengthening and had more cholestatic liver disease. Group 1 piglets had histological evident intestinal adaptation and 1.5-fold intestinal lengthening (P = 0.001). Conclusions: These novel piglet models of short bowel syndrome are the first to represent the full clinical spectrum of intestinal failure as observed in human neonates. By considering the impact of different short bowel anatomy on potential for adaptation and growth, these animal models are a significant advance. They permit evaluation of new therapies to promote intestinal adaptation and reduce complications, such as cholestasis.

The threonine requirement of sows increases in late gestation

Current AA recommendations for sows are to provide a fixed amount of AA intake throughout gestation based on the assumption that there is a constant demand for AA; however, the demand for nutrients changes from maternal lean tissue in early gestation to fetal and mammary growth in late gestation. The objective of this study was to determine the Thr requirement in early (d 35 to 53 and 25 to 55 for Exp. 1 and 2, respectively) and late (d 92 to 110 and 81 to 111 for Exp. 1 and 2, respectively) gestation using the indicator AA oxidation (IAAO) method with l-[1-(13)C]Phe as the tracer AA. A total of 14 multiparous sows were used: 6 in Exp. 1 and 8 in Exp. 2. Each sow received each of 6 diets in random order in both early and late gestation. A basal diet was formulated to contain Thr at 60% of the 1998 NRC recommendation in Exp. 1 and 20 and 60% of the 1998 NRC in Exp. 2 for early and late gestation, respectively. Crystalline l-Thr was added to create additional diets with approximately 10% incremental increases in Thr. Sows were placed in respiration chambers, and expired air and blood were collected every 30 min for 5.5 h. Tracer Phe [mg/(kg of BW·h)] was given orally over the last 4 h divided into eight 0.5-h meals. Expired air and plasma were measured for (13)CO(2) enrichment and free Thr concentration, respectively. Background (13)CO(2) was subtracted from plateau (13)CO(2) enrichment. Data were analyzed using a 2-phase nonlinear Mixed model. The overall litter size and litter weight were 13.5 ± 3.1 and 20.5 ± 3.9 kg, respectively. Based on IAAO, the Thr requirement in early gestation was 6.1 g/d (R(2) = 0.59, Exp. 1) and 5.0 g/d (R(2) = 0.71, Exp. 2). In late gestation, the Thr requirement based on IAAO was 13.6 g/d (R(2) = 0.60, Exp. 1) and 12.3 g/d (R(2) = 0.58, Exp. 2). Based on plasma Thr, the Thr requirement in early gestation was 7.0 g/d (R(2) = 0.90, Exp. 1) and 3.9 g/d (R(2) = 0.90, Exp. 2). In late gestation, the Thr requirement based on plasma Thr was 10.5 g/d (R(2) = 0.67, Exp. 2). There was a linear response to increasing Thr intake in late gestation in Exp. 1. Feeding a single amount of AA throughout gestation results in overfeeding AA in early gestation and underfeeding AA in late gestation. The 2-fold increase in Thr requirement in the last third of gestation suggests that phase feeding sows in gestation will more closely meet the demands for nutrients and that the requirement for essential AA in gestating sows should be re-evaluated in early and late gestation separately.

Exogenous glucagon-like peptide-2 improves outcomes of intestinal adaptation in a distal-intestinal resection neonatal piglet model of short bowel syndrome

Background:Endogenous glucagon-like peptide-2 (GLP-2) levels and intestinal adaptation are reduced in distal-intestinal resection animal models of short bowel syndrome (SBS) that lack remnant ileum. We hypothesized that exogenous GLP-2 would improve intestinal adaptation in a distal-intestinal resection neonatal piglet model of SBS.Methods:In all, 35 piglets were randomized to 2 treatment and 3 surgical groups: control (sham), 75% mid-intestinal resection (JI), and 75% distal-intestinal resection (JC). Parenteral nutrition (PN) commenced on day 1 and was weaned as enteral nutrition (EN) advanced. IV GLP-2 (11 nmol/kg/d) or saline was initiated on day 2. Piglets were maintained for 14 d. Clinical, functional, morphological, and histological outcomes were obtained.Results:JC-GLP-2 piglets had fewer days on PN (10.0 ± 0.6 vs. 13.8 ± 0.2), more days on EN (4.0 ± 0.6 vs. 0.2 ± 0.2), a higher percentage of EN at termination (92 ± 5 vs. 52 ± 10%), fewer days of diarrhea (8.0 ± 0.7 vs. 12.3 ± 0.4), increased intestinal length (19 ± 4 vs. −5 ± 3%), and deeper jejunal crypts (248 ± 21 vs. 172 ± 12 μm), compared with saline piglets.Conclusion:GLP-2 therapy improves clinical, morphological, and histological outcomes of intestinal adaptation in a distal-intestinal resection model of SBS. Since this anatomical subtype represents the majority of clinical cases of neonatal SBS, these results support a potential role for GLP-2 therapy in pediatric SBS.

Barley-derived β-glucans increases gut permeability, ex vivo epithelial cell binding to E. coli, and naïve T-cell proportions in weanling pigs1,2

Weaning in young animals is associated with an increased incidence of gastrointestinal infections. β-glucans exert numerous physiological effects, including altering immune function. The objective of this study was to determine the effects of feeding barley (Hordeum vulgare L.)-derived β-glucans on immune and intestinal function in weanling pigs (Sus scrofa). Thirty-one individually-housed Dutch Landrace pigs (21 d; initial BW, 6,298 ± 755 g) were weaned and fed a wheat-based diet (control) or a low (Lo-BG), medium (Med-BG), or high β-glucan-containing barley-based diet (Hi-BG) for 2 wk with 7 or 8 pigs/treatment. Intestinal segments were analyzed for permeability using Ussing chambers and K88 Escherichia coli adhesion to enterocytes was assessed ex vivo. Immune cells from mesenteric lymph nodes, peripheral blood, and Peyers patches were analyzed for lymphocyte subsets by indirect immunofluorescence and the ability to respond ex vivo to mitogens by (3)H-thymidine incorporation. Hematology and neutrophil function were determined by flow cytometry. Neutrophil burst, size, and granularity, lymphocyte proliferation, and B-cell distribution in peripheral blood lymphocytes, Peyers patches, and mesenteric lymph nodes were not affected by β-glucans content of the diet. The β-glucans content of the diet altered blood concentrations of erythrocytes and leukocytes, CD4, CD45RA, and CD8 blood cells (P < 0.05). In addition, feeding β-glucan resulted in increased (P < 0.05) percentage CD45RA positive cells in peripheral blood lymphocytes, Peyers patches, and mesenteric lymph nodes. Mannitol permeability and tissue conductance were increased (P < 0.05) in Hi-BG fed pigs compared with control pigs. Percentage maximum K88-E.coli binding was increased in proportion to the β-glucan content of the diet (P < 0.05). Although β-glucan feeding during the weaning period increased blood lymphocytes and the proportion of naïve T-cells, it also increased E. coli-enterocyte binding and intestinal permeability. β-glucan may alter immune and intestinal function of weaning pigs.

Effects of Polymeric Formula vs Elemental Formula in Neonatal Piglets With Short Bowel Syndrome

BACKGROUND Intestinal adaptation is important for recovery in short bowel syndrome (SBS). This process is dependent on the presence of enteral nutrition (EN) and trophic factors, such as glucagon-like peptide-2 (GLP-2). In clinical practice, elemental formula is often used to feed neonates with SBS, whereas animal studies suggest polymeric formula promotes better intestinal adaptation. In neonatal piglet models of SBS, with or without ileum, we compared the elemental with the polymeric formula, including the effect on endogenous GLP-2. MATERIALS AND METHODS Forty-eight piglets underwent 75% mid-intestinal resection with jejunoileal anastomosis, 75% distal-intestinal resection with jejunocolic anastomosis (JC), or sham without resection. Parenteral nutrition (PN) started postoperatively, tapering as EN was increased, according to clinical criteria, based on diarrhea and weight. Within groups, piglets were randomized to an isocaloric/isonitrogenous elemental (amino acid) or polymeric (intact protein) diet. Plasma GLP-2 and histology for adaptation were measured at 14 days. RESULTS Within both SBS and control groups, no difference in adaptation was observed according to diet. A difference was observed only within the JC piglet group with regard to clinical outcomes. In these piglets, compared with elemental formula, the polymeric formula was associated with more diarrhea ( P = .023) and longer duration of PN support (P = .047). CONCLUSION An overall benefit of the polymeric formula over the elemental formula on gut adaptation was not observed. Furthermore, SBS piglets without ileum had less ability to tolerate polymeric formula, contributing to more days of PN support.

Role of glucagon-like peptide-2 deficiency in neonatal short-bowel syndrome using neonatal piglets

Background:Short-bowel syndrome (SBS) is the most common cause of neonatal intestinal failure. Recovery requires intestinal adaptation, dependent on enteral nutrition (EN) and growth factors such as glucagon-like peptide–2 (GLP-2), which is secreted from L cells in the ileum. Neonatal SBS often results in loss of ileum; therefore, we hypothesized that without ileum, endogenous GLP-2 production would be inadequate to promote adaptation. We compared endogenous GLP-2 production and adaptation in neonatal animals with SBS, with and without ileum.Methods:Neonatal piglets (4–6 d) were randomized to 75% mid-intestinal resection, 75% distal-intestinal resection, or sham control without resection. Postoperatively, all piglets commenced parenteral nutrition (PN), tapering as EN was increased to maintain specific growth.Results:The resected SBS piglets developed intestinal failure, requiring a longer duration of PN support and experiencing fat malabsorption. The piglets without ileum were not able to wean from PN during the study and did not show adaptation, specifically growth in intestinal length or crypt hyperplasia on histology of the jejunum. Adaptation was observed in the resected SBS piglets with ileum, and these piglets also had an increased plasma GLP-2 level that was not observed in piglets without ileum.Conclusion:SBS piglets with ileum undergo adaptation associated with increased endogenous GLP-2 production. SBS piglets without ileum undergo limited adaptation and severe intestinal failure, requiring prolonged PN support. This appears to be related to a deficiency in endogenous GLP-2 production.

Dietary lysine requirement of sows increases in late gestation.

Current AA recommendations for sows are to provide a fixed amount of AA intake throughout gestation; however, the demand for nutrients changes from maternal lean tissue in early gestation (EG) to fetal and mammary growth in late gestation (LG). The objective of this study was to determine the Lys requirement in EG (d 24 to 45) and LG (d 86 to 110) using the indicator AA oxidation method with simultaneous determination of heat production. Each of 7 Large White × Landrace sows received 6 diets in random order in both EG and LG. Three semisynthetic diets (14.0 MJ ME/kg) based on corn were formulated and mixed to produce a basal diet (60% of 1998 NRC Lys requirement) and high diets for EG and LG (150% and 185% of 1998 NRC Lys requirements, respectively). The 6 test diets provided Lys intakes of 7.5 to 19.3 g/d in EG and 8.1 to 23.7 g/d in LG. Sows were placed in respiration chambers, and expired air and blood were collected every 30 min for 5.5 h. The tracer AA, l-[1-(13)C]Phe, was given orally at a rate of 2 mg/(kg BW ⋅ h) over the last 4 h, divided into 8, 0.5-h meals. Expired air was measured for (13)CO(2) enrichment, and plasma was measured for l-[1-(13)C]Phe enrichment and free Lys concentration. Background (13)CO(2) was subtracted from plateau (13)CO(2) enrichment. Requirements were determined using a 2-phase nonlinear model. Mean maternal BW gain in gestation (43.7 kg; pooled SE, 1.2 kg), litter size (14.6 total born piglets; pooled SE, 0.8), and litter weight (19.4 kg; pooled SE, 0.9 kg) did not differ between parities. Sow weight gain and BW was greater (P = 0.001) in LG than EG. Lysine requirement was 9.4 and 17.4 g/d in EG and LG, respectively. Phenylalanine retention in LG was maximized at a Lys intake of 17.7 g/d. Heat production was more (P = 0.069) and energy retention less (P = 0.019) in LG than EG. Energy retention in LG was not different from 0. Quantitative Phe kinetics in EG were not affected by Lys intake. In LG, Phe retention increased with Lys intake (P = 0.004), whereas Phe oxidation decreased (P = 0.005). The Lys requirement was determined to be less than current recommendations in EG and more than current recommendations in LG. To meet the change in requirements, diets with increased lysine content are needed in LG. Increasing the feed allowance in LG is necessary to maintain a positive energy balance throughout gestation.

Isoleucine requirement of pregnant sows

The objective of this study was to determine the Ile requirement in early (d 39 to 61) and late (d 89 to 109) pregnancy using the indicator AA oxidation method. The same 7 Large White × Landrace sows in their fourth parity were used in early and late pregnancy. Each sow received 6 diets based on corn, corn starch, and sugar in both early and late pregnancy at constant feed allowances (2.5 kg/d). Diets provided Ile at 20, 40, 60, 80, 100, and 120% of the Ile requirement (6.2 g/d based on the 1998 NRC) in early and 60, 80, 100, 140, 160, and 180% in late pregnancy. After determination of (13)C background in expired CO2 and plasma free Phe for 1.5 h when confined in respiration chambers, sows were fed the tracer, L[1-(13)C]Phe, a rate of 2.0 mg/(kg BW·h) over 4 h divided into eight 30-min meals. Expired CO2 and plasma free Phe were analyzed for (13)C enrichment above background. Requirements were determined as the breakpoint in 2-phase nonlinear models. Sow BW was 246.5 kg in early and 271.6 kg in late pregnancy. Daily gain of the 6 sows was similar in early (344 g/d) and late pregnancy (543 g/d). During pregnancy, sow maternal gain was 19.1 ± 4.4 kg and litters of 17.7 ± 0.8 piglets weighed 22.6 ± 0.9 kg at birth. The Ile requirement was 3.6 ± 1.2 g/d (P = 0.001) in early pregnancy with a Phe retention (-0.59 g/d) and energy retention (-0.31 MJ/d) that were not different from 0. This indicates that the fourth parity sows had requirements close to maintenance in early pregnancy. The Ile requirement in late pregnancy was 9.7 ± 1.9 g/d (P = 0.001) when sows retained 3.30 g/d of Phe and -1.45 MJ/d of energy. The greater Ile requirement in late pregnancy was probably caused by the increased conceptus growth after d 70 of pregnancy. Phenylalanine flux, oxidation, and nonoxidative disposal increased (P < 0.1) from early to late pregnancy, but body protein breakdown did not. Phenylalanine oxidation, nonoxidative disposal, and retention increased (P < 0.01) with increasing Ile intake in early pregnancy but were not affected by Ile intake in late pregnancy. Body protein breakdown did not respond to Ile intake in early or late pregnancy. Although energy retention was similar in early and late pregnancy, the respiratory quotient decreased (P = 0.047) from early (1.05) to late pregnancy (0.98), indicating lipid mobilization in late pregnancy when Ile was at or above the requirement. The results of this study show that the Ile requirement of sows increases from early to late pregnancy.

Protein intake but not feed intake affects dietary energy for finishing pigs

The effects of dietary protein and feeding levels on dietary metabolizable (ME) and net energy (NE) content were determined in 24 pigs, each offered two diets at 2.0 times the energetic maintenance requirement or for ad libitum intake between 55 and 95 kg body weight. Within feeding levels, pigs received, in random order, low-protein (LP; 11.2% CP, 0.61% lysine) or high-protein (HP; 20.2% CP, 0.61% lysine) diets of similar digestible energy content. Dietary NE was calculated from heat production based on 24-h indirect calorimetry following a 7-day N-balance period. Feed intake was greater for LP than HP when fed for ad libitum intake (p = 0.001). Protein level did not affect daily gain (p > 0.1) but HP improved gain: feed (p = 0.003). Dietary ME and NE were not significantly affected by feeding level but were decreased by high protein intake (p < 0.07). Reducing dietary protein reduced urinary energy losses and increased energy retention but did not affect heat production. The effect of dietary protein restriction was already evident on the ME level and carried over to a similar degree to the NE level because the utilization of ME was not affected by protein level. Dietary ME and NE decreased by 0.012 MJ/kg (p = 0.014) and 0.018 MJ/kg (p = 0.062), respectively, for each gram per day N intake. The results suggest that although there was an effect of protein level on NE, the greatest effect occurred at the level of ME. However, the prediction of both ME and NE may be improved by adopting energy values for dietary protein that changes with dietary protein content.

Hepatic ultrastructure in a neonatal piglet model of intestinal failure-associated liver disease (IFALD)

This study was designed to evaluate liver disease in neonatal piglets with surgical short bowel syndrome causing intestinal failure with partial parenteral nutrition dependence. The short bowel piglets had 75% surgical resection of distal small intestine, including all ileum and cecum, and were compared with sham controls, without resection, and to healthy sow-reared controls. After 18 days of combined parenteral and enteral nutrition in short bowel and sham piglets, liver tissue was collected for quantitative and semi-quantitative histological and ultrastructural evaluation. The short bowel piglets developed biochemical and histological cholestasis, not observed in sham and control piglets. Ultrastructural examination revealed bile canaliculus dilation with bile plugging, microvillus flattening and disappearance, but without abnormalities of the pericanalicular zone. Interestingly, these data are similar to bile canaliculus changes seen in human neonates with IFALD supporting an initial consideration of this model to elucidate the pathogenesis of IFALD.