Olexandr Fedkiv

An elemental diet fed, enteral or parenteral, does not support growth in young pigs with exocrine pancreatic insufficiency

BACKGROUND & AIMS Young individuals with exocrine pancreatic insufficiency (EPI) show growth reduction that can be reversed by dietary pancreatic enzyme supplementation. Here we investigated whether feeding an elemental diet could replace the growth-promoting effect of enzyme supplementation in EPI pigs. METHODS Weaned pigs with intact pancreas (control) or pancreatic duct-ligated (EPI pigs) were given a commercial pig feed, a fat-enriched diet, or an elemental diet, intragastrically and intravenously, with or without porcine pancreatin (Creon) supplementation for 1 week. RESULTS Control pigs, irrespective of receiving pig feed or an elemental diet, increased their body weight by 13.4-20.1%, while EPI pigs showed negligible weight gain. Giving a fat-enriched diet did not improve growth of the EPI pigs. However, if the EPI pigs were supplemented with pancreatin in combination with fat-enriched feed or the elemental diet, i.v., their body weight increased by 16.6 %and 8.5%, respectively. CONCLUSION Control pigs maintained normal growth, independently of the diet being given in polymeric or elemental form, while EPI pigs showed impaired growth when receiving the same diets without enzyme supplementation. Pancreatic juice and enzyme preparations, in addition to their digestive properties, also appear to affect nutrient assimilation and anabolism in young individuals.

Pancreatic and pancreatic-like microbial proteases accelerate gut maturation in neonatal rats.

Objectives Postnatal gut maturation in neonatal mammals, either at natural weaning or after precocious inducement, is coinciding with enhanced enzymes production by exocrine pancreas. Since the involvement of enzymes in gut functional maturation was overlooked, the present study aimed to investigate the role of enzymes in gut functional maturation using neonatal rats. Methods Suckling rats (Rattus norvegicus) were instagastrically gavaged with porcine pancreatic enzymes (Creon), microbial-derived amylase, protease, lipase and mixture thereof, while controls received α-lactalbumin or water once per day during 14–16 d of age. At 17 d of age the animals were euthanized and visceral organs were dissected, weighed and analyzed for structural and functional properties. For some of the rats, gavage with the macromolecular markers such as bovine serum albumin and bovine IgG was performed 3 hours prior to blood collection to assess the intestinal permeability. Results Gavage with the pancreatic or pancreatic-like enzymes resulted in stimulated gut growth, increased gastric acid secretion and switched intestinal disaccharidases, with decreased lactase and increased maltase and sucrase activities. The fetal-type vacuolated enterocytes were replaced by the adult-type in the distal intestine, and macromolecular transfer to the blood was declined. Enzyme exposure also promoted pancreas growth with increased amylase and trypsin production. These effects were confined to the proteases in a dose-dependent manner. Conclusion Feeding exogenous enzymes, containing proteases, induced precocious gut maturation in suckling rats. This suggests that luminal exposure to proteases by oral loading or, possibly, via enhanced pancreatic secretion involves in the gut maturation of young mammals.

Precocious gut maturation and immune cell expansion by single dose feeding the lectin phytohaemagglutinin to suckling rats.

The dietary lectin phytohaemagglutinin (PHA) induces gut growth and precocious maturation in suckling rats after mucosal binding. The present study investigated the dose range in which PHA provokes gut maturation and if it coincided with immune activation. Suckling rats, aged 14 d, were orogastrically fed a single increasing dose of PHA: 0 (control), 2, 10, 50 or 250 microg/g body weight (BW) in saline. The effect on gut, lymphoid organs and appearance of CD3+ (T-lymphocyte) and CD19+ (B-lymphocyte) cells in the small-intestinal mucosa was studied at 12 h (acute) and 3 d (late phase) after treatment. The low PHA doses (2 and 10 microg/g BW) induced intestinal hyperplasia without mucosal disarrangement but did not provoke gut maturation. Only the high PHA doses (50 and 250 microg/g BW) temporarily disturbed the intestinal mucosa with villi shortening and decrease in disaccharidase activities, and later after 3 d provoked precocious maturation, resulting in an increase in maltase and sucrase activities and decrease in lactase activity and disappearance of the fetal vacuolated enterocytes in the distal small intestine. Exposure to the high, but not to the low, PHA doses increased the number of mucosal CD19+ and CD3+ cells in the small intestine after 12 h, a finding also observed in untreated weaned rats aged 21-28 d. In conclusion, there was a dose-related effect of PHA on gastrointestinal growth and precocious maturation that coincided with a rapid expansion of mucosal B- and T-lymphocytes, indicating a possible involvement of the immune system in this process.

The effectiveness of enzymatic replacement therapy measured by turbidimetry and the lipaemic index in exocrine pancreatic insufficient young, growing pigs, fed a high-fat diet.

PURPOSE Conventionally, the management of exocrine pancreatic insufficiency (EPI) involves the consumption of a specific diet as well as the replacement of pancreatic enzymes, the effectiveness of which is usually measured by a classical method of blood analyses of non-esterified fatty acids (NEFA) and triglycerides (TG). Dietary supplementation with a pancreatic enzyme preparation (PEP), in conjunction with a high-fat diet, on growth performance, digestibility and absorption (analysed using turbidimetry) of dietary fat in pigs with EPI was investigated. MATERIALS/METHODS EPI was developed by surgical ligation of the pancreatic duct of six male pigs, 6 weeks of age. The pigs were fed a high fat diet (twice daily). A PEP containing 1800 mg entero-coated pancreatin was included in the high fat meals. Blood, urine and faecal samples were collected. The urine and faeces were analysed for dry matter, crude protein and fat content. The lipaemic index and plasma lipid profiles were assessed. RESULTS EPI completely stopped growth of the pigs. Treatment with PEP significantly increased (P<0.05) growth and body mass as well as the digestibility of dry matter and crude protein. PEP significantly improved the co-efficient of fat absorption, the lipaemic index (measured by turbidimetry methods) and caused significant changes in plasma nonesterified fatty acids and triglyceride concentrations. CONCLUSIONS The short term enzymatic replacement therapy together with a high fat meal has immediate beneficial effects on diet digestibility and on the growth retardation observed in EPI pigs. The turbidimetry method used to measure lipaemic index is a reliable, quick and efficient technique in measuring plasma lipid profiles and thus a good tool for assessing fat absorption.

A piglet with surgically induced exocrine pancreatic insufficiency as an animal model of newborns to study fat digestion.

The maldigestion and malabsorption of fat in infants fed milk formula results due to the minimal production of pancreatic lipase. Thus, to investigate lipid digestion and absorption and mimic the situation in newborns, a young porcine exocrine pancreatic insufficient (EPI) model was adapted and validated in the present study. A total of thirteen EPI pigs, aged 8 weeks old, were randomised into three groups and fed either a milk-based formula or a milk-based formula supplemented with either bacterial or fungal lipase. Digestion and absorption of fat was directly correlated with the addition of lipases as demonstrated by a 30% increase in the coefficient of fat absorption. In comparison to the control group, a 40 and 25% reduction in total fat content and 26 and 45% reduction in n-3 and n-6 fatty acid (FA) content in the stool was observed for lipases 1 and 2, respectively. Improved fat absorption was reflected in the blood levels of lipid parameters. During the experiment, only a very slight gain in body weight was observed in EPI piglets, which can be explained by the absence of pancreatic protease and amylase in the gastrointestinal tract. This is similar to newborn babies that have reduced physiological function of exocrine pancreas. In conclusion, we postulate that the EPI pig model fed with infant formula mimics the growth and lipid digestion and absorption in human neonates and can be used to elucidate further importance of fat and FA in the development and growth of newborns, as well as for testing novel formula compositions.

Effects on gut properties in exocrine pancreatic insufficient (EPI) pigs, being growth retarded due to pancreatic duct ligation at 7 weeks but not at 16 weeks of age.

PURPOSE Exocrine pancreatic insufficiency (EPI) induced in young pigs by pancreatic duct ligation (PDL) early after weaning result in total growth deprivation while it has little effect in somewhat older pigs. The main objective was to study effects of EPI on gut structure and function in littermate pigs underwent to PDL at different age. MATERIAL/METHODS Pigs, duct-ligated at either 7 (2 weeks post-weaning, PDL-7) or 16 weeks of age (PDL-16), and euthanized at an age of 21-23 weeks together with un-operated littermates were studied. The intestinal in vitro permeability was studied in separate PDL-pigs and compared to un-operated. RESULTS Morphometric analysis showed gut mucosal atrophy in the PDL-7 as compared to PDL-16 pigs, while no differences in mucosal disaccharidase activities. The intestinal permeability for different-sized markers was significantly increased in the PDL-pigs compared to the un-operated controls. Analyses of the intestinal digesta showed a total lack of pancreatic enzymes in all PDL-pigs, while instead new, as yet unidentified, enzyme-activities appeared. CONCLUSIONS All EPI-pigs, independent of age at PDL-operation, displayed adaptive gut changes, however the EPI-pigs operated early after weaning appeared more sensitive, probably related to their gut maturity and possibly explaining the growth arrest seen in these pigs.

Exogenous pancreatic-like enzymes are recovered in the gut and improve growth of exocrine pancreatic insufficient pigs

The exocrine pancreatic insufficient (EPI) pigs grow less due to different disturbances in feed digestion, absorption, and retention. Use of pancreatic-like enzymes of microbial origin in pigs may improve feed use and performance in slow-growing pigs. The aim was to study gut recovery and effectiveness of pancreatic-like enzymes of microbial origin supplementation on pig performance. Six male pigs 10 to 12 kg BW underwent pancreatic duct ligation surgery to induce total exocrine pancreatic insufficiency (EPI). Three cannulas to access the gastrointestinal tract content were installed in stomach, duodenum, and ileum in EPI pigs and in 3 control (healthy) pigs. One month after surgery, enzymes were given before feeding and digesta samples were collected for analyses. The BW of EPI pigs did not increase during 1 mo following surgery (11.7 vs. 11.6 kg BW); however, BW increased after 1 wk of enzyme supplementation (12.1 kg BW). Coefficient of fat and N absorption increased (P < 0.05) in EPI pigs after enzyme supplementation. Activity of amylase, lipase, and protease in chyme samples of EPI pigs was very low compared to controls. In EPI pigs after enzyme supplementation, amylase activity increased from 5.32 to 72.9 units/mL but remained lower than that of healthy pigs (162.7 units/mL). Lipase activity increased from 79.1 to 421.6 units/mL, which was similar to that of controls (507.3 units/mL). Proteolytic activity increased from 7.8 to 69.7 units/mL but still did not reach control pigs (164.3 units/mL). In conclusion, exogenous microbial enzymes mimic endogenous pancreatic enzymes being recovered along the lumen of the gastrointestinal tract. These enzymes might be a useful tool to stimulate growth of slower-growing pigs after the weaning period.

The effect of long-term lactobacilli (lactic acid bacteria) enteral treatment on the central nervous system of growing rats.

The aim of this study was to explore the relationship between consumption of large doses of lactic acid bacteria (LAB) and the behaviour and brain morphobiochemistry of normal growing rats. Four groups of rats were treated with LAB cultures twice daily for 6 months. The control group received 1 ml of saline per treatment, while two experimental groups received 1 ml of living bacteria (Lactobacillus plantarum and Lactobacillus fermentum, respectively) and the remaining group received a heat-treated (inactivated) L. fermentum culture. After 2 and 6 months of treatment, respectively, eight animals from each group were sacrificed, and specimens were taken for further analyses. The behaviour of the rats was evaluated five times in an open-field test at monthly intervals throughout the study. Lactobacilli treatment for 2 months induced changes in the motoric behaviour of the rats. The concentration of the astrocytesoluble and filament glial fibrillary acidic protein (GFAP) decreased in the posterior part of the hemispheres, including the thalamus, hippocampus and cortex of the rats treated with L. fermentum. A greater decrease in filament GFAP (up to 50%) was shown in the group receiving the live form of L. fermentum. In contrast, the GFAP in the live L. plantarum-treated group increased, showing elevated levels of the soluble and filament forms of GFAP in the posterior part of the hemispheres. A 60-66% decrease in the amount of the astrocyte-specific Ca-binding protein S-100b was shown in the posterior parts of the hemispheres and in the hindbrain of rats given LAB for 2 months. Prolonged feeding with LAB for 4 months up to full adulthood led to a further decrease in astrocyte reaction, reflected as an additional decrease in the amount of soluble GFAP and locomotor activity in all experimental groups. The changes in filament GFAP and S-100b appeared to disappear after prolonged feeding (total of 6 months) with LAB. In summary, LAB dietary treatment affected the ontogenetic development of the astrocytes, with the highest intensity observed in the early stages of rat development. It can be postulated that LAB treatment may play a preventive role in neurological diseases by decreasing astrocyte reaction and, consequently, lowering locomotor activity.

Stimulating effect of pancreatic-like enzymes on the development of the gastrointestinal tract in piglets.

Use of nutritional components from the milk and eventually from the solid feed relates to the growth and development of gastrointestinal tract (GIT). We studied the effect of pancreatic-like enzymes [porcine pancreatic enzymes (Creon) or microbial-derived amylase, protease, and lipase] on GIT morphology and lipid absorption in suckling piglets. Both enzyme preparations, in low or high dose, were fed via a stomach tube twice a day for 7 d starting at 8 d of age and controls received vehicle, n = 6. The day after treatments ended, lipid absorption was tested after which pigs were euthanized and GIT was examined. Enzyme cocktails, irrespective of their origin, increased (P < 0.001) triglyceride level in blood. Enzyme preparation affected (P < 0.001) small intestinal mucosal thickness, villi length, and crypt depth and (P < 0.01) mitotic division of enterocytes. In addition, the external administration of pancreatic enzymes stimulated pancreatic growth as observed by increased (P < 0.05) mitotic division of pancreatic cells. The study revealed that pancreatic or pancreatic-like enzymes of microbial origin administrated in the early postperinatal period enhance GIT development and may be used to better prepare the GIT of piglets for milk use and weaning.

Is hyperoxaluria in a porcine model of Roux-en-Y gastric bypass (RYGB) associated with exocrine pancreatic insufficiency?

1Department of Epizootiology and Clinic of Infectious Diseases, University of Life Sciences, Lublin, Poland Head of the Department: prof. Stanislaw Winiarczyk, PhD 2Department of Preclinical Veterinary Sciences, University of Life Sciences, Lublin, Poland Head of the Department: prof. Jose L. Valverde Piedra, PhD 3Department of Animal Physiology, University of Life Sciences, Lublin, Poland Head of the Department: prof. Iwona Puzio, PhD 4Department of Biology, Lund University, Lund, Sweden Head of the Department: prof. Christer Lofstedt, PhD 5R&D, SGPlus, Malmo, Sweden Chief Executive Officer: prof. Stefan G Pierzynowski, PhD 6General Surgery Department of the District Specialist Hospital, Lublin, Poland Head of the Department: Jerzy Mackiewicz, MD, PhD 7Department of Vitreoretinal Surgery, Medical University of Lublin, Poland Head of the Department: Andrzej Chrościcki, MD, PhD 8Department and Clinic of Animal Internal Diseases, University of Life Sciences, Lublin, Poland Head of the Department: Jacek Madany, MD, PhD 9Department of Human Anatomy, Laboratory of Biostructure, Medical University of Lublin, Poland Head of the Department: prof. Ryszard Maciejewski, MD, PhD 10Institute of Rural Health, Lublin, Poland Head of the Institute: prof. Iwona Bojar, MD, PhD 11Department of Cytology, Bogomoletz Institute of Physiology, Kiev, Ukraine Head of Department: prof. Galyna Skibo, MD, PhD