Effects of dose and formulation of carvacrol and thymol on bacteria and some functional traits of the gut in piglets after weaning

Two trials were conducted to study the effects of dose and formulation of carvacrol and thymol on bacterial counts, metabolites and functional traits of the gut in weaned piglets. In the first experiment (Exp. I), 25 piglets (28 d, 6.59 ± 0.48 kg BW) were allocated to five dietary treatments: a control diet, or the same diet supplemented with either carvacrol or thymol at doses of 500 and 2000 mg kg^?1. In the second experiment (Exp. II), 35 piglets (28 d, 7.99 ± 0.73 kg BW) were assigned to seven dietary treatments: the same control diet as in Exp. I, or this diet supplemented with thymol in one of three formulations (on celite, on alphacel or microencapsulated) at doses of 500 and 2000 mg kg^?1. At 11/12 days post-weaning piglets were euthanised, and digesta from stomach, proximal and distal small intestine were sampled for bacteriological and biochemical analysis. Small intestinal tissue was sampled for histo-morphological determinations. In none of the experiments or sections of the gut was the number of bacteria lowered by the carvacrol or thymol supplementation. In Exp. I, the villus/crypt ratio at the distal small intestine for the experimental diets (1.30–1.32) was higher than for the control diet (1.24) (p < 0.05). Thymol fed animals in Exp. II had a lower number of intra-epithelial lymphocytes at the proximal (p < 0.05) and at the distal (p < 0.1) small intestine as compared to control animals. Mean concentration of the active ingredient in the stomach and proximal small intestine for the 2000 mg kg^?1 carvacrol diet was 521 and 5 mg kg^?1 fresh digesta, respectively, and for the 2000 mg kg^?1 thymol diets it ranged between 475 and 647 and between 13 and 24 mg kg^?1 fresh digesta, respectively. Cumulative absorption in the proximal small intestine was higher than 90% for all treatments and was not affected by formulation type. These data suggest that carvacrol and thymol can improve gut health, but evidence for clear antimicrobial effects towards the major culturable bacteria of the pig foregut is limited.     

Intestinal concentrations of free and encapsulated dietary medium-chain fatty acids and effects on gastric microbial ecology and bacterial metabolic products in the digestive tract of piglets

The influence of low dietary levels of free and encapsulated medium-chain fatty acids on their concentrations in the digesta, the gastric microbial ecology and bacterial metabolic products in the gastrointestinal tract (GIT) in weaned piglets was studied. Starting after weaning, 36 piglets were fed a diet without (Control) or with medium-chain fatty acids uncoated (MCFA) or coated with vegetable fat and lecithin (MCFAc). After 4 weeks, the animals were killed, and digesta from the stomach and different sections of the GIT were collected. The concentrations of caprylic (p < 0.001) and capric (p = 0.001) acids were higher in the stomachs of piglets fed diets MCFA and MCFAc compared to the Control group. The concentrations dropped rapidly along the GIT, regardless of encapsulation, and tended to be higher in groups MCFA and MCFAc compared to the Control. Compared to the Control group, ingestion of diet MCFAc led to an increase in the number of eubacteria (p = 0.001), enterobacteriaceae (p < 0.001), clostridial clusters I (p = 0.001) and IV (p = 0.019), Lactobacillus johnsonii (p < 0.001) and Lactobacillus amylovorus (p = 0.001) in gastric contents. A similar trend was seen with diet MCFA. Relative concentrations of short-chain fatty acids were characterised by lower propionic acid levels (p = 0.045), numerically (p < 0.1) higher acetic, lower n-butyric and i-valeric acid concentrations in the small intestine. Lactic acid concentrations were not significantly changed in the GIT, but ammonia concentrations increased (p < 0.001) in the distal small intestine in the MCFA and MCFAc groups. In conclusion, medium-chain fatty acids affected microbial ecology parameters in the gastric contents and bacterial metabolites in the small intestine. At low dietary levels, medium-chain fatty acids may be regarded as modulators of the gastric microbiota in weaned piglets.     

Intestinal concentrations of free and encapsulated dietary medium-chain fatty acids and effects on gastric microbial ecology and bacterial metabolic products in the digestive tract of piglets

The influence of low dietary levels of free and encapsulated medium-chain fatty acids on their concentrations in the digesta, the gastric microbial ecology and bacterial metabolic products in the gastrointestinal tract (GIT) in weaned piglets was studied. Starting after weaning, 36 piglets were fed a diet without (Control) or with medium-chain fatty acids uncoated (MCFA) or coated with vegetable fat and lecithin (MCFAc). After 4 weeks, the animals were killed, and digesta from the stomach and different sections of the GIT were collected. The concentrations of caprylic (p < 0.001) and capric (p = 0.001) acids were higher in the stomachs of piglets fed diets MCFA and MCFAc compared to the Control group. The concentrations dropped rapidly along the GIT, regardless of encapsulation, and tended to be higher in groups MCFA and MCFAc compared to the Control. Compared to the Control group, ingestion of diet MCFAc led to an increase in the number of eubacteria (p = 0.001), enterobacteriaceae (p < 0.001), clostridial clusters I (p = 0.001) and IV (p = 0.019), Lactobacillus johnsonii (p < 0.001) and Lactobacillus amylovorus (p = 0.001) in gastric contents. A similar trend was seen with diet MCFA. Relative concentrations of short-chain fatty acids were characterised by lower propionic acid levels (p = 0.045), numerically (p < 0.1) higher acetic, lower n-butyric and i-valeric acid concentrations in the small intestine. Lactic acid concentrations were not significantly changed in the GIT, but ammonia concentrations increased (p < 0.001) in the distal small intestine in the MCFA and MCFAc groups. In conclusion, medium-chain fatty acids affected microbial ecology parameters in the gastric contents and bacterial metabolites in the small intestine. At low dietary levels, medium-chain fatty acids may be regarded as modulators of the gastric microbiota in weaned piglets.     

Impact of in-feed sodium butyrate or sodium heptanoate protected with medium-chain fatty acids on gut health in weaned piglets challenged with Escherichia coli F4+

ABSTRACT

Short and medium-chain fatty acids (SCFA and MCFA, respectively) are commonly used as feed additives in piglets to promote health and prevent post-weaning diarrhoea. Considering that the mechanism and site of action of these fatty acids can differ, a combined supplementation could result in a synergistic action. Considering this, it was aimed to assess the potential of two new in-feed additives based on butyrate or heptanoate, protected with sodium salts of MCFA from coconut distillates, against enterotoxigenic Escherichia coli (ETEC) F4⁺ using an experimental disease model. Two independent trials were performed in 48 early-weaned piglets fed a control diet (CTR) or a diet supplemented with MCFA-protected sodium butyrate (BUT+; Trial 1) or sodium heptanoate (HPT+; Trial 2). After 1 week of adaptation, piglets were challenged with a single oral inoculum of ETEC F4⁺ (minimum 1.4 · 10⁹ cfu). One animal per pen was euthanised on days 4 and 8 post-inoculation (PI) and the following variables assessed: growth performance, clinical signs, gut fermentation, intestinal morphology, inflammatory mediators, pathogen excretion and colon microbiota. None of the additives recovered growth performance or reduced diarrhoea when compared to the respective negative controls. However, both elicited different responses against ETEC F4⁺. The BUT+ additive did not lead to reduce E. coli F4 colonisation but enterobacterial counts and goblet cell numbers in the ileum were increased on day 8 PI and this followed higher serum TNF-α concentrations on day 4 PI. The Firmicutes:Bacteroidetes ratio was nevertheless increased. Findings in the HPT+ treatment trial included fewer animals featuring E. coli F4 in the colon and reduced Enterobacteriaceae (determined by 16S RNA sequencing) on day 4 PI. In addition, while goblet cell numbers were lower on day 8 PI, total SCFA levels were reduced in the colon. Results indicate the efficacy of MCFA-protected heptanoate against ETEC F4⁺ and emphasise the potential trophic effect of MCFA-protected butyrate on the intestinal epithelium likely reinforcing the gut barrier.     

Intestinal development and growth performance of early-weaned piglets fed a low-threonine diet

High dietary threonine extraction by the digestive tract suggests that threonine contributes to maintain gut integrity. The aims of this study were to investigate the intestine development and the growth performance of early-weaned piglets pair-fed either a control well-balanced (C: 9.3 g threonine/kg diet) or a low-threonine diet (LT: 6.5 g threonine/kg diet) for 2 weeks. As expected, LT piglets presented lower plasma free threonine compared with C piglets (118 v. 356 ± 12 μmol/l, P < 0.001). Dietary threonine supply altered neither growth performance nor growth of the intestine and of the other portal-drained viscera (stomach, spleen and pancreas). Nevertheless, villus height was reduced in the ileum of the LT piglets compared with C piglets (446 v. 714 ± 74 μm, P < 0.05). This was also associated with a decrease in crypt width (P < 0.05) and villus height-to-crypt depth ratio (P < 0.05). Whereas maltase and lactase activities did not change between the two groups, aminopeptidase nitrogen activity was decreased in the ileum of LT piglets (269 v. 374 ± 27 IU/mg protein, P < 0.05). The number of mucin-containing goblet cells was not modified in the ileum and in the proximal part of the large intestine of the LT piglets compared with the C piglets. In conclusion, despite no alteration of intestinal growth, villus hypotrophy associated with a reduction of aminopeptidase nitrogen activity suggest an alteration of the structure of the ileum in early-weaned piglets fed a diet supplying inadequate dietary threonine.     

The gut morphology of the African ice rat,<Emphasis Type="Italic"> Otomys sloggetti robertsi</Emphasis>, shows adaptations to cold environments and sex-specific seasonal variation

We studied the gut morphology of the ice rat Otomys sloggetti robertsi, a non-hibernating murid rodent endemic to the sub-alpine and alpine regions of the southern African Drakensberg and Maluti mountains. The gut structure of O. s. robertsi is well adapted for a high fibre, herbivorous diet, as is the case with other members of its subfamily Otomyinae. Despite the broad similarity in gross gut morphology with mesic- and arid-occurring otomyines, O. s. robertsi has a larger small intestine, caecum, stomach volume and parts of the colon, which we suggest are adaptations for increased energy uptake and/or poor diet quality in alpine environments. However, O. s. robertsi has a smaller larger intestine than other otomyines, perhaps because it occupies a mesic habitat. Seasonal sexual differences occurred, with females increasing dimensions of the stomach, small intestine length, caecum, and large intestine in summer. Sexual asymmetry in gut morphology may be related to increased energy requirements of females during pregnancy and lactation, indicating phenotypic plasticity in response to poor quality vegetation and a shorter growing season in alpine habitats.Communicated by G. Heldmaier     

Increased diet viscosity by oat β-glucans decreases the passage rate of liquids in the stomach and affects digesta physicochemical properties in growing pigs

Rheological properties of digesta play a role in digesta passage kinetics through the gastrointestinal tract, in turn affecting nutrient absorption kinetics. Therefore, we studied the effects of diet viscosity on digesta passage and physicochemical properties in pigs. Twenty male growing pigs (35 kg body weight at the start) were assigned to one of five diets with increasing dietary concentrations of β-glucans (BG; from 0 % to 10 %), in exchange for maize starch. After a 17-day adaptation period, pigs were euthanised and the mean retention time (MRT) of digesta solids (TiO₂) and liquids (Cr-EDTA) in the stomach, and proximal and distal half of the small intestine was quantified. In the stomach, the MRT of liquids, but not of solids, increased when dietary BG level increased (6 min per % dietary BG, P = 0.008 and R² = 0.35). Concomitantly, stomach DM content (5 g/kg per % dietary BG, P < 0.001 and R² = 0.53) and apparent digesta viscosity (56 Pa × s at 1/s shear rate per % dietary BG, P = 0.003 and R² = 0.41) decreased. In the proximal half of the small intestine, no effects of dietary BG level were observed. In the distal half of the small intestine, water-binding capacity (WBC) of digesta increased (0.11 g/g digesta DM per % dietary BG, P = 0.028 and R² = 0.24) and starch digestibility decreased (0.3% per % dietary BG, P = 0.034 and R² = 0.23) when dietary BG level increased. In the colon, apparent digesta viscosity at 45/s shear rate increased (0.1 Pa × s per % dietary BG, P = 0.03 and R² = 0.24) in the proximal half of the colon, and digesta WBC increased (0.06 g/g digesta DM per % dietary BG, P = 0.024 and R² = 0.26) in the distal half of the colon when dietary BG level increased. To conclude, increasing dietary BG level caused the MRT of liquids, but not that of solids, to increase in the stomach, resulting in reduced separation of the solid and liquid digesta fractions. This caused dilution of the stomach content and reduction in digesta viscosity when dietary BG levels increased. Effects of dietary BG level on physicochemical properties in the proximal small intestine were absent and may have been due to a low DM content. The WBC of digesta in the distal small intestine and colon increased when dietary BG level increased, as did apparent digesta viscosity in the proximal colon. This likely reflects the concentration of BG in digesta when moving through the gastrointestinal tract.     

The effects of copper, iron and zinc on digestive enzyme activity in the hybrid tilapia Oreochromis niloticus (L.) ×Oreochromis aureus (Steindachner)

The present experiment was conducted to study effects of Cu, Fe and Zn on activities of digestive enzymes of the hybrid tilapia Oreochromis niloticus×Oreochromis aureus. The acidic protease activities increased 65·5 and 55·1% by addition of homogenates of digesta‐containing stomach with copper (75 mg l⁻¹) and zinc (50 mg l⁻¹) respectively. Addition of Cu and Zn increased the activities of protease in the hepatopancreas homogenates by 132·7 and 38·1% respectively, and reduced the activity of protease in the digesta‐containing intestine homogenates by 11·0 and 13·8% respectively. Addition of Fe (50 mg l⁻¹) increased the acidic protease activity by 96·7% but did not alter the activities of protease in the intestine and hepatopancreas. Addition of Cu markedly inhibited activities of amylase in intestine and hepatopancreas homogenates, while Zn addition showed no effects. Addition of Fe reduced activities of amylase in the intestine homogenates by 47·9% but had no effect on amylase activities in the hepatopancreas. When Cu (75 mg kg⁻¹), Fe (50 mg kg⁻¹) and Zn (50 mg kg⁻¹) were supplemented to basal diet for 3 weeks, the activities of amylase in hepatopancreas homogenates increased 125·3, 215·6 and 70·0%, respectively, the activities of amylase in intestine increased 79·8, 74·6 and 48·5%, respectively, and the activities of lipase in intestine increased 90·5, 149·8 and 84·0%, respectively. Supplementation of Cu, Fe or Zn into diet had no effects on activity of protease in all digestive organs. Therefore, the results suggest that effects of Cu, Fe and Zn on activity of digestive enzymes in vitro were different from those seen in vivo, and that the positive effects of Cu, Fe and Zn supplemented to fish diet would be valuable information for formulating fish feed.     

The yellow-flowered species of Cuphea (Lythraceae), including three new taxa

Among the predominantly purple- and red-flowered species of the genusCuphea, only seven yellow floral tube and/or yellow-petaled members in three sections were known. We add here two new species and one new variety.Cuphea congesta from Venezuela andC. splendida var.viridiflava from Bolivia are new to sect.Melvilla.Cuphea xanthopetala from Brazil is described in sect.Euandra and expands the presence of this feature to a fourth section of the genus. A key is provided to allCuphea with yellow floral tubes and/or yellow petals to expedite determinations when these exceptional character states are present.     

In vitro fermentability and physicochemical properties of fibre substrates and their effect on bacteriological and morphological characteristics of the gastrointestinal tract of newly weaned piglets

Abstract

Fermentability of fibre has a great impact on the bacterial flora along the gastrointestinal tract of newly weaned piglets. Therefore, this parameter was determined by incubating in vitro different fibre substrates (chicory roots, sugar beet pulp, wheat bran and corn cobs) with contents of jejunum or caecum sampled from slaughtered pigs. Incubating with small intestinal contents, lactic acid was the only fermentation product. Fermentability was highest for chicory roots, followed by wheat bran and sugar beet pulp, while corn cobs were not fermented. Based on SCFA formed in the incubations with caecal contents, ranking of the fermentability of the fibre substrates was in the same order. The effect of adding different fibre substrates to diets of newly weaned piglets on bacteriological and morphological aspects of the gastrointestinal tract was also investigated. In Experiment 1 three groups of five piglets, weaned at four weeks of age, received a control feed (C), C supplemented with corn cobs (50 g/kg) or with chicory roots (20 g/kg). In Experiment 2, diet C was supplemented with sugar beet pulp (120 g/kg) or with wheat bran (75 g/kg). After three weeks animals were euthanized and digesta were sampled from stomach, proximal and distal jejunum, caecum and colon. Furthermore, mucosal scrapings were prepared and tissue samples were taken from jejunum, caecum and colon. Viscosity was determined for jejunal, caecal and colon contents. Corn cobs in the feed increased the number of total bacteria, lactobacilli and bifidobacteria in the stomach and proximal duodenum, while a decreased count of streptococci in distal jejunum contents was noted. Chicory roots increased the counts of Escherichia coli in the distal jejunum and on the mucosa, while sugar beet pulp decreased the number of lactobacilli on the mucosa only. Wheat bran seemed to increase the count of E. coli in jejunal digesta and on the mucosa, and also the number of lactobacilli in the stomach and jejunum. Bifidobacterial numbers were increased but only in the proximal part of the jejunum. Fibre substrates affected the concentration of lactate and SCFA in different parts of the intestinal tract. Feeding corn cobs increased villus length in the proximal jejunum by 13%. The number of intra-epithelial lymphocytes in the villous epithelium of proximal and distal jejunum was decreased by corn cobs and chicory roots supplementation while beet pulp and wheat bran had the opposite effect. In Experiment 1, apoptotic index of the mucosa of the distal jejunum was very low and decreased when corn cobs were fed. Mitotic index in the crypts was only affected by the wheat bran diet and a small decrease was noted. It was concluded that the fermentability of fibre was not an ideal criterion for predicting its effects on the flora. The effect of fibres on viscosity of digesta was negligible probably explaining the lack of clear and consistent influences on the intestinal mucosa.     

Influence of Tribulus terrestris extract on the survival and histopathology of Oreochromis mossambicus (Peters, 1852) fry before and after Streptococcus iniae infection

The aim of this study was to determine the effects of Tribulus terrestris extract (TT) on growth performance, disease resistance and histopathological changes in intestine and liver tissues of Oreochromis mossambicus (Peters, 1852) first‐feeding fry before and after exposure to Streptococcus iniae. Five isonitrogenous and isocaloric diets were formulated to contain 0 (control), 200, 400, 600, and 800 mg kg^?1 TT. After feeding for 45 days, fish were infected with S. iniae and mortalities recorded. Final weight, weight gain and SGR of tilapia fry fed the 400 mg kg^?1 TT diet were significantly greater than that of control diet. In the challenge experiment, the best survival rate was obtained with 400 mg kg^?1 TT supplementation. Infection by S. iniae appeared to have a negative effect on histopathological findings and outcome than did TT‐800 used alone. However, administration of TT (200 or 400 mg extract kg^?1) resulted in overall improvement in the intestine and liver histopathology, emphasizing the protective potential of TT. The present study suggests the protective potential of TT in alleviating intestinal and hepatic damage that can occur after a S. iniae infection. It was concluded that 400 mg kg^?1 TT can enhance growth and disease resistance during first–feeding of O. mossambicus fry. This suggests that TT may be an alternative to antibiotics in controlling streptococcal disease in tilapia culture.     

Digestive tract morphology and digestion in the wombats (Marsupialia: Vombatidae)

Summary Wombats consume grasses and sedges which are often highly fibrous. The morphology of the digestive tract and the sequence of digestion were studied in two species of wombats from contrasting habitats: Vombatus ursinus from mesic habitats and Lasiorhinus latifrons from xeric regions. Studies were performed on wild wombats consuming their natural winter diets, and on captive wombats fed a high-fibre pelleted straw diet. Vombatus had a shorter digestive tract (9.2 vs 12.5 times body length) of greater capacity (wet contents 17.9 vs 13.7% body weight) than Lasiorhinus. The most capacious region of the digestive tract was the proximal colon (62–79% of contents). The proportional length and surface area of the proximal colon were greater in Vombatus, but those of the distal colon were greater in Lasiorhinus. These digestive morphologies may reflect adaptations for greater capacity and longer retention of digesta in Vombatus, but greater absorption and lower faecal water loss in Lasiorhinus. Apparent digestion along the digestive tract was estimated by reference to lignin. The proximal colon was the principal site of fibre and dry matter digestion, whereas nitrogen was mainly digested in the small intestine. Depot fats in captive wombats were highly unsaturated and reflected those in the diet. Therefore, lipids, proteins and soluble carbohydrates in the plant cell contents were digested and absorbed in the stomach and small intestine. Conversely, dietary fibre was probably retained and digested by microbial fermentation along the proximal colon.Abbreviations ADF acid detergent fibre - DM dry matter - NDF neutral detergent fibre - SD standard deviation     

Neurotrophin receptors and enteric neuronal development during metamorphosis in the amphibian<Emphasis Type="Italic"> Xenopus laevis</Emphasis>

During metamorphosis, the frog intestine goes through a dramatic shortening with extensive apoptosis and regeneration in the epithelial layer and connective tissue. Our aim was to study changes in the enteric nervous system represented by one inhibitory (vasoactive intestinal polypeptide; VIP) and one excitatory (substance P, neurokinin A; SP/NKA) nerve population and concomitant changes in neurotrophin receptor occurrence during this development in the gut of Xenopus laevis adults and tadpoles at different stages of metamorphosis (NF stages 57–66). Sections were incubated with antibodies against the neurotrophin Trk receptors and p75^NTR, and the neurotransmitters VIP and SP/NKA. Trk-immunoreactive nerves increased dramatically but transiently in number during early metamorphic climax. Nerves immunoreactive for p75^NTR were present throughout the gut, decreased in number in the middle intestine during climax, and increased in the large intestine during late metamorphosis. The percentage of VIP-immunoreactive nerves did not change during metamorphosis. SP/NKA-immunoreactive nerves were first apparent at NF stages 61–62 in the middle intestine and increased in the stomach and large intestine during metamorphosis. Endocrine cells expressing SP/NKA increased in number in stomach, proximal, and middle intestine during metamorphic climax. Thus, neurotrophin receptors are expressed transiently in neurons of the enteric nervous system during metamorphosis in Xenopus laevis and SP/NKA innervation is more abundant in the intestine of the postmetamorphic frog than in the tadpole.This study was supported by grants from the Swedish Research Council to S. Holmgren     

Dietary total aromatic amino acid requirement and tyrosine replacement value for phenylalanine in Indian major carp: Cirrhinus mrigala (Hamilton) fingerlings

Two 8‐week growth trials were conducted to determine total aromatic amino acid requirement and tyrosine replacement value for phenylalanine in Cirrhinus mrigala fingerlings. To determine the phenylalanine requirement, 20 fish were randomly stocked in triplicate groups in 55‐L indoor polyvinyl flow‐through circular tanks and fed six experimental diets containing graded levels of phenylalanine (5.0, 7.5, 10.0, 12.5, 15.0 and 17.5 g kg^?1, dry diet) with 10 g kg^?1 tyrosine. Maximum weight gain (287%), best FCR (1.44) and PER (1.74) occurred at 12.5 g kg^?1 dietary phenylalanine. Quadratic regression analysis of weight gain, FCR and PER data indicated phenylalanine requirement at 13.5, 12.9 and 12.7 g kg^?1 of dry diet, respectively. Protein deposition was significantly (P < 0.05) higher at 12.5 g kg^?1 dietary phenylalanine. Based on the above results, phenylalanine requirement of C. mrigala is recommended at 13.0 g kg^?1 of dry diet, corresponding to 32.5 g kg^?1 of protein. On the basis of the above requirement, a second experiment with a similar design was conducted using six diets containing graded levels of tyrosine (2.1, 4.0, 6.0, 8.0, 10.0 and 12.0 g kg^?1) with 13.0 g kg^?1 phenylalanine fixed in all diets to determine the phenylalanine replacement value with that of tyrosine. Maximum weight gain (315%), best FCR (1.47) and PER (1.69) was at 8.0 g kg^?1 dietary tyrosine. Quadratic regression analysis of weight gain, FCR and PER data indicated tyrosine requirement at 9.0, 8.4 and 8.2 g kg^?1 of dry diet, respectively. Protein deposition was significantly (P < 0.05) higher at 8.0 g kg^?1 dietary tyrosine. On the basis of the above results, 8.5 g kg^?1 tyrosine, corresponding to 21.3 g kg^?1 of protein, is taken as the optimum requirement and the replacement value is 39.53% on a weight and 36% on a molar basis. Thus, the total aromatic amino acid requirement is 21.5 g kg^?1 of diet, corresponding to 53.8 g kg^?1 of protein for optimum C. mrigala growth.     

Epidermal growth factor improves intestinal morphology by stimulating proliferation and differentiation of enterocytes and mTOR signaling pathway in weaning piglets

Epidermal growth factor(EGF) has been shown to improve piglet intestinal morphology and epithelial recovery. In an attempt to further understand the mechanisms behind these improvements, this study tested the hypothesis that dietary EGF may affect intestinal morphology by stimulating the proliferation and differentiation of enterocytes in weaning piglets. In piglets receiving200 μg kg–1 EGF, crypt depth and villus height increased(P0.05). Adding 400 μg kg–1 EGF increased villus height-to-crypt depth ratio(P0.05), but reduced crypt depth(P0.05). Dietary supplementation with 200 μg kg–1 EGF significantly increased the number of Ki67-positive cells(P0.01) and tended to increase the mRNA level of proliferating cell nuclear antigen(P0.10).However, this supplementation decreased the expression level of intestinal fatty acid-binding protein(P0.05). Piglets fed with400 μg kg–1 EGF had an increased mRNA level of intestinal alkaline phosphatase(P0.05). The phosphorylation of m TOR(mammalian target of rapamycin) was observed in the 200 μg kg–1 EGF group. These results suggest that dietary supplementation with a low level of EGF improved piglet intestinal morphology through stimulating the proliferation and differentiation of enterocytes, and the mTOR signaling pathway may partly be involved in this process.     

A moderate threonine deficiency differently affects protein metabolism in tissues of early-weaned piglets

A moderate threonine deficiency may affect differently tissue protein metabolism. In this study, we compared protein metabolism in the small and large intestines, the liver, and the carcass of piglets (Sus scrofa) pair-fed either a control well-balanced diet (C: 9.3 g threonine/kg diet) or a low threonine diet (LT: 6.5 g threonine/kg diet) for 2 weeks. In the small intestine, the LT diet did not modify protein deposition, fractional protein synthesis rate (K_S) and AA protein composition. Ubiquitin mRNA level, a component of the ubiquitin-dependent proteolytic pathway, was significantly decreased in the jejunum of the LT piglets. Protein deposition measured in the carcass and the colon, and K_S measured in the semitendinosus muscle and the colon, did not differ between LT and C piglets. Nevertheless, in these compartments, threonine content was reduced indicating deposition of proteins less rich in threonine. In the liver, protein retention was reduced, K_S was increased and AA protein composition was modified in the LT compared to the C piglets. In conclusion compared to the other compartments, small intestinal protein metabolism seems to be less sensitive to a moderate dietary threonine deficiency. This indicates that dietary threonine extraction by the small intestine may reduce threonine availability for the other tissues when young piglets were fed a diet marginally deficient in threonine.     

Effect of dietary L‐malic acid supplementation on growth,feed utilization and digestive function of juvenile GIFT tilapia Oreochromis niloticus (Linnaeus, 1758)

Two feeding trials (FTs) were conducted in 2013 and 2014, respectively, to determine the optimal L‐malic acid (LMA) level for juvenile GIFT (Genetically Improved Farmed Tilapia) Oreochromis niloticus. Except for the LMA level, the FT1 and FT2 had a similar diet formulation. In FT1, LMA was included at 0 (basal diet), 1, 4, 8, 16 and 32 g kg^?1, respectively. After 20 weeks, daily weight gain and feed conversion ratio were improved but not differentiated with 1–8 g kg^?1 LMA. Further increasing the LMA supply initially decreased the feed intake (16 g kg^?1), and then decreased both feed intake and feed utilization (32 g kg^?1), thus impairing the fish growth. FT2 was subsequently conducted with a smaller LMA range (0, 0.5, 1, 2, 4 and 8 g kg^?1, respectively) but was unfortunately terminated at the end of 8 weeks because 20% of the fish were badly injured during weighing. Unexpectedly, growth and feed utilization were still improved but not differentiated with 0.5–8 g kg^?1 LMA. In FT2, beneficial effects of LMA inclusion on the digestive function (pepsin, foregut amylase and foregut lipase), the activities of serum lysozyme and hepatic superoxide dismutase, and liver lipid peroxidation (malondialdehyde concentration) were found. Taking the results of FT1 and FT2 together, it could be concluded that dietary LMA supplementation at low concentrations (0.5–8 g kg^?1) could improve growth and feed utilization, but excess LMA (≥16 g kg^?1) might compromise feed intake and/or feed utilization, thus impairing fish growth. To reduce feed costs in commercial practice, 0.5 g kg^?1 LMA is recommended in the feed of juvenile GIFT tilapia based on the results of this study.     

the influence of lentinus edodes (shiitake mushroom) preparations on bacteriological and morphological aspects of the small intestine in piglets

Among substances intended to replace growth promoting antibiotics in pig nutrition, non-digestible oligosaccharides or polysaccharides could be potential alternative compounds. Therefore, the influence of β-1,3-1,6 glucans on bacteriological, biochemical and morphological aspects of the small intestine in weaned piglets was investigated. As sources of β-glucans, Lentinan (extract of Lentinus edodes mycelium) or dried L. edodes mycelium were added to the diet. Four homogenous groups of 5 newly weaned piglets (4 weeks of age) received one of four diets: control diet (C), C supplemented with Avilamycin (50?mg/kg, positive control), C supplemented with 0.1% of Lentinan and C supplemented with 5% of dried L. edodes mycelium powder. A first group of 10 piglets was euthanized after 11 days and the remaining 10 on day 12 of the experiment. The gastrointestinal tract was divided in segments and samples taken from digesta (stomach, proximal and distal jejunum, caecum), mucosal scrapings (jejunum) and ring shaped tissue samples (1?cm) of proximal and distal jejunum. Bacterial counts were made with digesta and mucosal samples, and short-chain fatty acids (SCFA), lactic acid and ammonia concentrations were determined. Tissue samples of both jejunal sites were embedded in paraffin wax for morphometrical (villus length, crypt depth) and histological observations (numbers of intraepithelial lymphocytes (IEL), goblet cells, apoptotic enterocytes on villi, mitotic cells in crypts). Only the diet containing 5% of dried L. edodes consistently resulted in lower viable counts (ca. 1?–?2 log₁₀ CFU) of total bacteria, E. coli, streptococci and lactic acid bacteria, and luminal and mucosal effects agreed very well. With this diet, acetate and butyrate concentrations in the distal jejunum were doubled, which is favourable in view of the trophic effect on enterocytes and colonocytes. Villus length (V) was increased with both diets containing β-glucans while crypt depth (C) was not altered, but V/C was higher. IEL counts were decreased by both diets although bacterial numbers, which is only one parameter of bacterial load, were only diminished with the L. edodes feed. The three supplemented feeds lowered the number of apoptotic enterocytes on the villi, but these numbers were very low (control diet?:?44 cells per 100 villi), making clear interpretation difficult. The mitotic index was slightly lower with the L. edodes feed, although not statistically significant. Decreased viable counts observed with the latter diet is a favourable effect as it is accepted that a lower bacterial load causes lower turnover rates of the intestinal epithelial cells, while there is also less competition for specific substrates. A higher V/C ratio, a smaller number of IEL in the epithelium and a lower apoptotic index also indicate slower turnover rate of the mucosa when Lentinan and L. edodes diets were fed. The inconsistent effects observed with Lentinan were probably due to the low amount added to the diet. It should be taken into account that the influence of L. edodes mycelium powder was more likely due to the presence of antibacterial compounds (eg. lenthionine, lentinamycin, terpenoids, polyphenols), rather than to an immunostimulating action of β-glucans with increased release of IgA onto the mucosa surface.     

Cholecystokinin and peptide YY are released by fat in either proximal or distal small intestine in dogs

We tested the hypothesis that the release of cholecystokinin (CCK) and peptide YY (PYY) may be independent of the region of the small intestine exposed to fat. In five dogs equipped with duodenal and midgut fistulas, the small intestine was compartmentalized so that fat was confined to either the proximal or distal one-half of the gut. Plasma CCK and PYY levels were measured by radioimmunoassay and compared by the square root of the area under the curve (sqrt AUC), representing the plasma peptide concentration over time. CCK was released similarly whether fat was delivered into the proximal (69.9+/-4.7 pM) or distal (71.0+/-5.5 pM) gut, but significantly more CCK (88.9+/-5.6 pM; p<0.05) was released when both the proximal and distal gut were perfused simultaneously with fat. PYY was released similarly whether fat was delivered into the proximal (34.9+/-2.6 pM) or distal (40.0+/-1.2 pM) gut or both (38.6+/-2.2 pM). We conclude that CCK and PYY are released by fat in either the proximal or distal one-half of the small intestine.     

Do herbivorous minnows have “plug-flow reactor” guts? Evidence from digestive enzyme activities,gastrointestinal fermentation,and luminal nutrient concentrations

Few investigations have empirically analyzed fish gut function in the context of chemical reactor models. In this study, digestive enzyme activities, levels of gastrointestinal fermentation products [short chain fatty acids (SCFA)], luminal nutrient concentrations, and the mass of gut contents were measured along the digestive tract in herbivorous and carnivorous minnows to ascertain whether their guts function as “plug-flow reactors” (PFRs). Four of the species, Campostoma anomalum, C. ornatum, C. oligolepis, and C. pauciradii, are members of a monophyletic herbivorous clade, whereas the fifth species, Nocomis micropogon, is a carnivore from an adjacent carnivorous clade. In the context of a PFR model, the activities of amylase, trypsin and lipase, and the concentrations of glucose, protein, and lipid were predicted to decrease moving from the proximal to the distal intestine. I found support for this as these enzyme activities and nutrient concentrations generally decreased moving distally along the intestine of the four Campostoma species. Furthermore, gut content mass and the low SCFA concentrations did not change (increase or decrease) along the gut of any species. Combined with a previous investigation suggesting that species of Campostoma have rapid gut throughput rates, the data presented here generally support Campostoma as having guts that function as PFRs. The carnivorous N. micropogon showed some differences in the measured parameters, which were interpreted in the contexts of intake and retention time to suggest that PFR function breaks down in this carnivorous species.