Effect of dipeptide on intestinal peptide transporter 1 gene expression: An evaluation using primary cultured chicken intestinal epithelial cells

Peptide transporter 1 (PepT1) is a transporter responsible for absorbing dipeptide and tripeptide in enterocytes and is upregulated by dipeptide in mammals. It has not been certain whether intestinal PepT1 expression is responsive to dipeptides in chickens because of the lack of in vitro study using the cultured enterocytes. This study established a primary culture model of chicken intestinal epithelial cells (IECs) in two-dimensional monolayer culture using collagen gel by which the response of chicken PepT1 gene expression to dipeptide stimuli was evaluated. The cultured chicken IECs showed the epithelial-like morphology attached in a patch-manner and exhibited positive expression of cytokeratin and epithelial cadherin, specific marker proteins of epithelial cells. Moreover, the chicken IECs exhibited the gene expression of intestinal cell type-specific marker, villin1, mucin 2, and chromogranin A, suggesting that the cultured IECs were composed of enterocytes as well as goblet and enteroendocrine cells. PepT1 gene expression was significantly upregulated by synthetic dipeptide, glycyl-l-glutamine, in the cultured IECs. From the results, we herein suggested that dipeptide is a factor upregulating PepT1 gene expression in chicken IECs.     

Nutritional value and physiological effects of soya-free diets fed to rats during growth and reproduction

The objective of the study was to assess the effects of substitution milk and egg for soya products in breeding diets for rats, with concomitant decrease of the dietary protein level and supplementation with amino acids. Soya-containing (S) and two soya-free (NS and NSA) diets were evaluated as protein and energy sources, and their effects on reproductive performance during two cycles, and on the quality of the offspring were assessed. Organ weights were registered in females and blood parameters were determined in males. In the offspring males from S and NS groups, plasma LH, testosterone and prolactin levels were measured on the 22nd and the 60th day of life. The S diet contained more protein of smaller concentration of methionine and cystine and lower biological value than both NS and NSA diets and promoted similar post-weaning growth rate, similar body weight changes of dams during gestation and lactation and slightly lower mating efficiency. Within each reproductive cycle, the number and individual and total body weight of newborn and weanling pups did not differ but in two cycles mean number of neonates per litter and mean litter weight were significantly lower on S than on NSA diet. Plasma concentration of hormones did not differ in 22-day-old offspring males while in the older ones LH and prolactin levels were higher in animals fed on S than on NS diet. It is concluded that replacing soya protein by milk and egg protein with concomitant lowering dietary protein level and amino acid supplementation does not impair the growth rate and tends to improve reproductive performance. Feeding soya-free vs. soya-containing diets differentiates hormonal status of young males.     

The effects of feeding diets containing potato protein concentrate on reproductive performance of rats and quality of the offspring

Two diets containing either soybean meal (SBM) or SBM and potato protein concentrate (PPC) in proportion of protein 1:1 (diet BS and BPP, respectively), were fed each to 22 females and 22 males from the age of 4-5 weeks throughout two reproductive cycles. A commercial closed-formula SNF diet containing unknown amount of potato protein was fed till the end of the first reproductive cycle. All three diets were also fed during 21 days following weaning, each to 10 offspring males born in the first cycle. The solanidine glycoalkaloids (GA) content in PPC was 2316 mg/kg DM. Feeding diets containing potato protein did not affect pregnancy rate, number and conformation of neonates and number of pups weaned (in all litters number of neonates left with dams being reduced to eight) but it depressed their body weight (weight of weaners on BPP diet only in the first cycle). Growth rate of the offspring males did not differ but feed intake was lower and feed conversion was higher on BPP than on BS and SNF diets. Feeding of BPP diet to offspring did not affect mass of body organs but increased blood urea and albumin concentrations while feeding SNF diet increased relative weight of small intestine and liver and raised blood concentrations of alanine amino transferase, albumin, protein, urea and cholesterol. In dams sacrificed after two cycles only relative weight of uterus and ovaries were lower in females fed on BPP than on BS diet. It is concluded that feeding moderate amounts (6.4%) of high-GA PPC does not impair reproductive performance, does not induce malformation of pups but depresses foetal and postnatal growth rate till weaning but not after weaning.     

The effect of pectin on amino acid digestibility and digesta viscosity, motility and morphology of the small intestine, and on N-balance and performance of young pigs

Two experiments were carried out using a balanced feed mixture composed of wheat, corn, soyabean meals, casein, and crystalline amino acids (AA) and supplemented with 0, 4, or 8% of apple pectin. The daily dietary allowance provided pigs with the same level of nutrients, apart from the pectin. One experiment was conducted on pigs (within 25 to 40 kg BW) with the post valve T-caecum (PVTC) cannula to determine AA digestibility and ileal digesta viscosity. In the second experiment, the N-balance was measured twice on male pigs at about 20 and 28 kg BW using 6 animals per diet and feed intake and body weight were measured twice during the 28 d. After 40 d of feeding the diets, the pigs were sacrificed and the small intestines were weighed and sampled. The added pectin decreased standardized ileal AA digestibility (on average up to 5% dig. units) and increased digesta viscosity from about 1 to 88 mPa s. The higher pectin level did not affect motility of the duodenum and mid-jejunum, measured as responses to electrical field stimulation and to acetylcholine. However, the response of the duodenum to electrical field stimulation was after feeding the diet with 4% pectin increased compared with other diets. Pectin supplementation did not alter the weight and the length of the small intestine, but induced changes in the intestinal morphology. Muscle layer width increased significantly in the duodenum, the mid-jejunum, and the ileum while villi length increased in the duodenum and ileum. The addition of pectin decreased N retention and increased the F/G ratio but did not affect significantly the ADG of the pigs.     

Dietary protein level affects in vitro peristaltic reflex in the small intestine of young pigs

The aim of the study was to investigate the effect of dietary protein level on contractility of the small intestine in young pigs. Animals (about 15 kg BW initially) were fed during 40 days with isoenergetic diets containing 16, 18, or 20% of crude protein. Higher protein levels in the diets as well as similar concentration of essential amino acids were attained by substitution of corn starch with wheat gluten and crystalline amino acids. At about 40 kg BW, the animals were sacrificed and the longitudinal strips of duodenum and mid-jejunum were taken for analysis of smooth muscle mechanical activity in vitro. Spontaneous activity, responses to electrical field stimulation (EFS), and responses to acetylcholine (ACh) were recorded. Frequency and amplitude of spontaneous contractions were similar in all groups, both in the duodenum and the mid-jejunum. In the duodenum, responses to EFS and ACh were the same irrespective of the dietary protein level. Responses to EFS in the mid-jejunum were significantly enhanced in the animals receiving the diet with 20% protein; however, the response to ACh was not changed. The mechanism of neuronal response may involve primary afferent pathways “sensing” contents in the lumen of the intestine through enterochromaffin cells. The enteric neuronal pathways may also be activated by a higher level of free amino acid(s) or by dietary peptides and proteins.     

Prevalence and consequence of subacute ruminal acidosis in Polish dairy herds

The aim of this study was to estimate the prevalence of subacute ruminal acidosis (SARA) in Polish high‐yielding dairy herds. Also, the relationships between the chemical composition of the diet and the feed particle size, ruminal pH and the occurrence of this metabolic disease and the fermentation profile were determined. Rumen fluid samples were obtained from a total of 213 cows from nine dairy herds (≥20 cows per herd) via rumenocentesis. Almost 14% (30/213) of cows as acidotic (pH<5.6) were found, which is indicative of SARA. Moreover, 44% of the herds were classified as SARA‐positive and 56% as SARA‐negative. Results of the current study suggested that the physically effective NDF (NDF_>1.18 mm)‐to‐starch ratio could be better indices than peNDF_>1.18 mm to preventing the occurrence of SARA, and their level should not be lower than 1.00. Also, linear negative relationships between rumen fluid pH and concentration of propionate, valerate and total VFA were determined. According to the herd's SARA status and rumen fluid biochemical indices, there were significant differences between the pH of rumen fluid (p ≤ 0.01), concentrations of acetate (p ≤ 0.05), propionate (p ≤ 0.05), n‐butyrate (p ≤ 0.01), n‐valerate (p ≤ 0.01), the sum of VFA (p ≤ 0.01) and ammonia (p ≤ 0.05) in SARA‐positive herds compared to SARA‐negative herds. The better understanding the strategy of ruminant nutrition to coordinate energy conversion and the role of the ruminal pH in regulating N‐NH₃ production, absorption through rumen mucosa, urea secretion, the more successfully we can utilize these processes with due recognition of animal needs and welfare, as well as prevention of SARA occurrence.     

Goblet cells and mucus layer in the gut of young pigs: Response to dietary contents of threonine and non‐essential amino acids

Mucins secreted by goblet cells (GC) are the major components of mucus layer coating and protecting gut epithelium. The study aimed at determining the effect of non‐essential amino acids (NEAA) and threonine (Thr) levels on GC number and mucus layer thickness measured in different parts of the gut. A two‐factorial experiment was conducted on 72 pigs (initial BW 12.5 kg) using as factors: Thr level (5.1, 5.7, 6.3 and 6.9 g standardized ileal digestible [SID] Thr/kg) and wheat gluten (WG) level used as a source of NEAA (20.4, 40.4 and 60.4 g WG protein in WG20, WG40 and WG60 diets respectively). All diets covered the requirement for essential AA, except for Thr, and they were fed to six pigs for 20 days. Thr level affected only the count of GC containing acidic mucins in the proximal colon, which was higher in pigs fed a diet with 5.1 g SID Thr/kg in comparison with diet containing 6.3 g SID Thr/kg. In the villi, WG40 and/or WG60 increased the GC number containing acidic mucins and lowered that with neutral mucins in the middle jejunum. In the crypts, higher WG levels decreased the GC number with acidic mucins in the duodenum, ileum and proximal colon and increased that with neutral and mixed mucins in the duodenum and ileum. The mucus layer was thicker in the proximal and middle jejunum in WG20 pigs, whereas in the proximal colon it was thicker in WG60 pigs. The lowest GC count was found in the middle jejunum and the highest in the duodenum and proximal colon. The mucus layer was the thinnest in the proximal and middle jejunum and the thickest in the ileum. The results did not show the negative effect of a moderate Thr deficiency and the positive effect of non‐essential AA supplementation on GC functions.     

Proteome changes in renal cortex and medulla induced by dietary supplementation with inulin‐type fructans in growing pigs

The aim of the study was to evaluate the influence of dietary supplementation with inulin extract from chicory root and dried chicory root on the protein profile of the renal cortex and medulla of growing pigs. The experiment was carried out on renal cortex and medulla tissue collected from 24 50‐day‐old PIC x Penarlan P76 crossbred piglets (males). Animals were divided into three dietary groups (n = 8) and fed with a control diet, diet supplemented with 2% inulin extract from chicory root and a diet supplemented with 4% dried chicory root. Kidney samples were collected after 40 days of feeding, and renal cortex and medulla proteins were separated by two‐dimensional electrophoresis. Protein identification was performed using MALDI‐TOF mass spectrometry. The diet supplemented with 2% chicory inulin induced significant expression changes of 20 and 26 protein spots in the renal cortex and medulla respectively. Supplementation with 4% dried chicory root triggered changes in the expression of 44 and 24 proteins in the renal cortex and medulla respectively. Both forms of chicory inulin‐type fructans effectively affected the expression of proteins involved in energy metabolism, heat shock proteins and other chaperones, cytoskeletal and cytoskeleton‐related proteins, as well as other proteins. Additionally, changes in transferrin abundance in both experimental groups suggested the significance of chicory fructan supplementation for iron absorption and bioavailability. In conclusion, 2% inulin extract from chicory root and 4% dried chicory root exerted a similar effect on changes in renal protein expression; however, more pronounced alterations were induced by dried chicory root. Nevertheless, further studies are needed for better understanding the mechanism underlying the effect of chicory inulin‐type fructans and their fermentation end products on the kidneys of growing pigs.     

Modulation of ruminal and intestinal fermentation by medicinal plants and zinc from different sources

Two experiments were conducted on sheep to determine the effect of dietary supplementation with zinc and a medicinal plant mixture on haematological parameters and microbial activity in the rumen and large intestine. In Experiment 1, 24 male lambs were randomly divided into four groups: One group was fed an unsupplemented basal diet (control), and three groups were fed a diet supplemented with 70 mg Zn/kg diet in the form of Zn sulphate (ZnSO₄), a Zn‐chelate of glycine hydrate (Zn‐Gly) or a Zn‐proteinate (Zn‐Pro), for five months. The ruminal content was collected separately from each lamb, and batch cultures of ruminal fluid were incubated in vitro with mixture of medicinal plants (Mix) with different roughage:concentrate ratios (800:200 and 400:600, w/w). Bioactive compounds in Mix were quantified by UPLC/MS/MS. In Experiment 2, four sheep were fed a diet consisting of meadow hay and barley grain (400:600, w/w), with Zn‐Gly (70 mg Zn/kg diet), Mix (10% replacement of meadow hay) or Zn‐Gly and Mix (Zn‐Gly‐Mix) as supplements in a Latin square design. Mix decreased total gas (p < 0.001) and methane (p < 0.01) production in vitro. In Experiment 1, caecal isobutyrate and isovalerate concentrations varied among the dietary treatments (p < 0.01). The isovalerate concentration of the zinc‐supplemented groups in the distal colon was higher (p < 0.001) compared with the control. In Experiment 2, the molar proportion of isobutyrate was the highest in the faeces of the sheep fed the diet with Zn‐Gly‐Mix (p < 0.01). The plasma zinc concentration was higher in the groups fed a diet supplemented with zinc (p < 0.001). The haematological profile and antioxidant status did not differ between the dietary groups (p > 0.05). The diets containing medicinal plants and organic zinc thus helped to modulate the characteristics of fermentation in ruminants.     

Growth performance and physiological parameters of conventional and specified pathogen‐free rats fed autoclaved diets with different protein sources

The effects of feeding autoclaved commercial SSNIFF (SN) diet and diets containing soya bean (S) and casein (C) to growing conventional (CON) and specified pathogen‐free (SPF) rats were determined. Diets S, C and SN, autoclaved at 121 °C during 20 min (T1), at 134 °C during 10 min (T2) and non‐autoclaved (T0), were fed during four weeks, each to 8 CON males and 8 females of mean initial body weight 56 g, kept individually. Diets S, C and SN, autoclaved at T1, were fed during two months, each to 20 SPF males and 20 females of mean initial body weight 58 g, kept in group of 5 animals per cage. In CON rats, autoclaving did not affect feed intake and weight gain, decreased thyroid and stomach weight, increased caecal tissue and digesta weight, and concentrations of isobutyric, isovaleric and valeric acid in caecal digesta. Among biochemical blood parameters, autoclaving decreased only total protein concentration and aspartate aminotransferase activity. Feeding C diet resulted in lower feed intake and weight gain in CON and SPF males. Diet affected organ weights and the greatest differences were found in rats on SN diet for weights of stomach, caecum and female reproductive organs. Diet affected concentration of all short‐chain fatty acids, pH and weight of caecal digesta, the most important being the greatest butyric acid concentration on SN diet and isoacids on C diet. It is concluded that autoclaving of both soya‐containing and soya‐free diets does not affect negatively animal performance and physiology.