Canine Mucosal Artificial Colon: development of a new colonic in vitro model adapted to dog sizes.

Differences in dog breed sizes are an important determinant of variations in digestive physiology, mainly related to the large intestine. In vitro gut models are increasingly used as alternatives to animal experiments for technical, cost, societal, and regulatory reasons. Up to now, only one in vitro model of the canine colon incorporates the dynamics of different canine gut regions, yet no adaptations exist to reproduce size-related digestive parameters. To address this limitation, we developed a new model of the canine colon, the CANIne Mucosal ARtificial COLon (CANIM-ARCOL), simulating main physiochemical (pH, transit time, anaerobiosis), nutritional (ileal effluent composition), and microbial (lumen and mucus-associated microbiota) parameters of this ecosystem and adapted to three dog sizes (i.e., small under 10 kg, medium 10-30 kg, and large over 30 kg). To validate the new model regarding microbiota composition and activities, in vitro fermentations were performed in bioreactors inoculated with stools from 13 dogs (4 small, 5 medium, and 4 large). After a stabilization period, microbiota profiles clearly clustered depending on dog size. Bacteroidota and Firmicutes abundances were positively correlated with dog size both in vitro and in vivo, while opposite trends were observed for Actinobacteria and Proteobacteria. As observed in vivo, microbial activity also increased with dog size in vitro, as evidenced from gas production, short-chain fatty acids, ammonia, and bile acid dehydroxylation. In line with the 3R regulation, CANIM-ARCOL could be a relevant platform to assess bilateral interactions between food and pharma compounds and gut microbiota, capturing inter-individual or breed variabilities. KEY POINTS: • CANIM-ARCOL integrates main canine physicochemical and microbial colonic parameters • Gut microbiota associated to different dog sizes is accurately maintained in vitro • The model can help to move toward personalized approach considering dog body weight.

Supplementation with nitrate only modestly affects lipid and glucose metabolism in genetic and dietary-induced murine models of obesity.

To gain a better understanding of how nitrate may affect carbohydrate and lipid metabolism, female wild-type mice were fed a high-fat, high-fructose diet supplemented with either 0, 400, or 800 mg nitrate/kg diet for 28 days. Additionally, obese female db/db mice were fed a 5% fat diet supplemented with the same levels and source of nitrate. Nitrate decreased the sodium-dependent uptake of glucose by ileal mucosa in wild-type mice. Moreover, nitrate significantly decreased triglyceride content and mRNA expression levels of Pparγ in liver and Glut4 in skeletal muscle. Oral glucose tolerance as well as plasma cholesterol, triglyceride, insulin, leptin, glucose and the activity of ALT did not significantly differ between experimental groups but was higher in db/db mice than in wild-type mice. Nitrate changed liver fatty acid composition and mRNA levels of Fads only slightly. Further hepatic genes encoding proteins involved in lipid and carbohydrate metabolism were not significantly different between the three groups. Biomarkers of inflammation and autophagy in the liver were not affected by the different dietary treatments. Overall, the present data suggest that short-term dietary supplementation with inorganic nitrate has only modest effects on carbohydrate and lipid metabolism in genetic and dietary-induced mouse models of obesity.

Differential action of TGR5 agonists on GLP-2 secretion and promotion of intestinal adaptation in a piglet short bowel model.

Enteroendocrine L cells and glucagon-like peptide 2 (GLP-2) secretion are activated in the intestinal adaptation process following bowel resection in patients with short bowel syndrome. We hypothesized that enteral activation of Takeda G protein-coupled receptor 5 (TGR5), expressed in enteroendocrine L cells, could augment endogenous GLP-2 secretion and the intestinal adaptation response. Our aim was to assess the efficacy of different TGR5 agonists to stimulate GLP-2 secretion and intestinal adaptation in a piglet short-bowel model. In study 1, parenterally fed neonatal pigs (n = 6/group) were gavaged with vehicle, olive extract (OE; 10 or 50 mg/kg), or ursolic acid (UA; 10 mg/kg), and plasma GLP-2 was measured for 6 h. In study 2, neonatal pigs (n = 6-8/group) were subjected to transection or 80% mid-small intestine resection and, after 2 days, assigned to treatments for 10 days as follows: 1) transection + vehicle (sham), 2) resection + vehicle (SBS), 3) resection + 30 mg UA (SBS + UA), and 4) resection + 180 mg/kg OE (SBS + OE). We measured plasma GLP-2, intestinal histology, cell proliferation, and gene expression, as well as whole body citrulline-arginine kinetics and bile acid profiles. In study 1, GLP-2 secretion was increased by UA and tended to be increased by OE. In study 2, SBS alone, but not additional treatment with either TGR5 agonist, resulted in increased mucosal thickness and crypt cell proliferation in remnant jejunum and ileum sections. SBS increased biliary and ileal concentration of bile acids and expression of inflammatory and farnesoid X receptor target genes, but these measures were suppressed by UA treatment. In conclusion, UA is an effective oral GLP-2 secretagogue in parenterally fed pigs but is not capable of augmenting GLP-2 secretion or the intestinal adaptation response after massive small bowel resection. NEW & NOTEWORTHY Therapeutic activation of endogenous glucagon-like peptide 2 (GLP-2) secretion is a promising strategy to improve intestinal adaptation in patients with short bowel syndrome. This study in neonatal pigs showed that oral supplementation with a selective Takeda G protein-coupled receptor 5 (TGR5) agonist is an effective approach to increase GLP-2 secretion. The results warrant further study to establish a more potent oral TGR5 agonist that can effectively improve intestinal adaptation in pediatric patients with SBS.

Developmental regulation of the intestinal FGF19 system in domestic pigs.

Fibroblast growth factor-19 (FGF19) is an emerging endocrine factor involved in the regulation of bile acid homeostasis and energy metabolism in rodents and humans. In pigs, however, the FGF19 system remains largely unexplored. This study was designed to investigate the developmental regulation of the FGF19 system in domestic pigs. Samples of intestinal sections, liver, and plasma were collected from 24 pigs ( n = 6) at four developmental stages (birth, preweaning, postweaning, and adulthood). In the intestine, expression of the farnesoid X receptor (FXR) and FGF19 showed a congruent time- and region-dependent regulation, beginning soon after birth to achieve maximal expression in ileum during adulthood. The same temporal pattern was followed by the circulating concentration of FGF19, and these changes were accompanied by a time-related increase in the ileal proportion of bile acids that potently activate FXR. Conversely, genes belonging to the FGF19 signaling machinery achieved maximal expression in the small intestine at birth to decrease sharply afterward. In the liver, gene expression of FGF19 receptors and enzymes involved in bile acid biosynthesis paralleled after-birth changes in plasma concentration of this enterokine and attained a maximum during postweaning when plasma FGF19 was the lowest. Although detectable at birth, the hepatic expression of genes belonging to the bile acid-FXR-FGF19 pathway was low before the onset of enteral feeding. In summary, the porcine FGF19 system is present from birth, operative before the onset of enteral feeding, and regulated in a temporal and section-specific manner. NEW & NOTEWORTHY Fibroblast growth factor-19 (FGF19) is an emerging endocrine factor. The domestic pig is a translational model of value in biomedical research. We show for the first time that in pigs the intestinal FGF19 system is present from birth, operative before the onset of enteral feeding, and regulated in a temporal and section-specific manner. This work identifies pigs as a suitable model for investigating the implications of FGF19 signaling within and beyond the gut-liver axis.

n-3 Fatty acids combined with flavan-3-ols prevent steatosis and liver injury in a murine model of NAFLD.

Non-alcoholic fatty liver disease (NAFLD) affects 25% of adults and at present no licensed medication has been approved. Despite its complex patho-physiology, dietary strategies aiming at delaying or preventing NAFLD have taken a reductionist approach, examining the impact of single components. Accumulating evidence suggests that n-3 LC-PUFAs are efficacious in regulating lipogenesis and fatty acid oxidation. In addition, plant derived flavonoids are also emerging as a dietary strategy for NAFLD prevention, with efficacy attributed to their insulin sensitising and indirect antioxidant effects. Based on knowledge of their complementary molecular targets, we aimed to demonstrate that the combination of n-3 LC-PUFA (n-3) and flavan-3-ols (FLAV) prevents NAFLD. In a high-fat high-fructose (HF/HFr) fed C57Bl/6J mouse model, the independent and interactive impact of n-3 and FLAV on histologically defined NAFLD, insulin sensitivity, weight gain, intestinal and hepatic gene expression, intestinal bile acids were examined. Only the combination of FLAV and n-3 (FLAVn-3) prevented steatosis as evidenced by a strong reduction in hepatocyte ballooning. While FLAV reduced body (-28-30%), adipose tissue (-45-50%) weights and serum insulin (-22-25%) as observed following an intra-peritoneal glucose tolerance test, n-3 downregulated the expression of Srebf1 and the lipogenic genes (Acaca, Fasn). Significant impacts of interventions on intestinal bile acid metabolism, farnesoid X receptor (Fxr) signalling in the intestine and liver, and hepatic expression of fatty acid transporters (Fabp4, Vldlr, Cd36) were also evident. FLAVn-3 may be a novel intervention for NAFLD. Future research should aim to demonstrate its efficacy in the prevention and treatment of human NAFLD.

Olive oil bioactive compounds increase body weight, and improve gut health and integrity in gilthead sea bream (Sparus aurata).

An olive oil bioactive extract (OBE) rich in bioactive compounds like polyphenols, triterpenic acids, long-chain fatty alcohols, unsaturated hydrocarbons, tocopherols and sterols was tested (0, 0·08, 0·17, 0·42 and 0·73 % OBE) in diets fed to sea bream (Sparus aurata) (initial weight: 5·4 (sd 1·2) g) during a 90-d trial (four replicates). Fish fed diets containing 0·17 and 0·42 % OBE were 5 % heavier (61·1 (sd 1·6) and 60·3 (sd 1·1) g, respectively) than those of the control group (57·0 (sd 0·7) g), although feed conversion ratio and specific feed intake did not vary. There were no differences in lipid peroxidation (LPO) levels, catalase, glutathione reductase and glutathione S-transferase activities in the intestine and liver, although there was a tendency of lower intestinal and hepatic LPO levels in fish fed OBE diets. No differences in villus size were found among treatments, whereas goblet cell density in the control group was on average14·3 % lower than in fish fed OBE diets. The transcriptomic profiling of intestinal markers, covering different biological functions like (i) cell differentiation and proliferation, (ii) intestinal permeability, (iii) enterocyte mass and epithelial damage, (iv) IL and cytokines, (v) pathogen recognition receptors and (vi) mitochondria function, indicated that among the eighty-eight evaluated genes, twenty-nine were differentially expressed (0·17 % OBE diet), suggesting that the additive has the potential of improving the condition and defensive role of the intestine by enhancing the maturation of enterocytes, reducing oxidative stress, improving the integrity of the intestinal epithelium and enhancing the intestinal innate immune function, as gene expression data indicated.

Cromolyn-mediated improvement of intestinal barrier function is associated with enhanced piglet performance after weaning.

BACKGROUND: Previous work showed that weaning stress causes gut barrier dysfunction partly by triggering the release of corticotropin releasing factor (CRF) and thereby inducing the degranulation of intestinal mast cell (MC). This study investigated the hypothesis that attenuating the weaning-induced activation of the CRF-MC axis via administration of a MC stabilizing agent (cromolyn) may improve gut permeability and piglet performance after weaning. RESULTS: To test the hypothesis twenty piglets were weaned (20 ± 1.0 d of age; 6.4 ± 0.4 kg of BW) and injected intraperitoneally with saline (control, n = 10) or 20 mg/kg BW of sodium cromolyn (cromolyn, n = 10) at - 0.5, 8 and 16 h relative to weaning. Piglets were housed individually and fed ad libitum a pre-starter diet from one to 15 d post-weaning followed by a starter diet until the end of the study on d 36. Cromolyn improved intestinal permeability as indicated by the reduced recovery of cobalt and mannitol in plasma samples. Cromolyn treated pigs consumed more feed (369 vs. 313 g/d; P < 0.009), gained more BW (283 vs. 238 g/d; P < 0.006), and grew more efficiently (0.60 vs. 0.40; P < 0.042) than their control counterparts. As a result, cromolyn treated pigs were 1.4 kg heavier than those in the control group by d 36 after weaning (16.5 vs. 17.9 kg; P < 0.002). CONCLUSIONS: In agreement with our hypothesis, present data indicate that the cromolyn-mediated improvement of intestinal permeability is associated with enhanced pig performance after weaning.

Bile acid mediated effects on gut integrity and performance of early-weaned piglets.

BACKGROUND: Early weaning (EW) results in a transient period of impaired integrity of the intestinal mucosa that may be associated with reduced plasma concentration of glucagon-like peptide-(GLP) 2. We have previously shown that intragastric infusion of chenodeoxycholic acid (CDC) increases circulating GLP-2 in early-weaned piglets. The aim of this study was to expand previous work to establish whether feeding piglets a cereal-based diet supplemented with CDC can improve gut integrity and animal performance immediately after EW. A cohort of 36 piglets weaned at 20 days of age, 6.2 ± 0.34 kg of body weight (BW) were randomly assigned (n = 18) to receive a standard prestarter diet or the same diet supplemented with 60 mg of CDC per kg of initial BW for ad libitum intake until day 14 postweaning. Thereafter, all pigs were fed the same untreated starter diet for 21 days until the end of the study on day 35. On days 1, 7 and 14 blood samples were collected from 6 pigs per treatment to measure plasma GLP-2. On day 15, 6 pigs per treatment were euthanized to obtain intestinal tissue samples for later histological and gene expression analyses. RESULTS: Supplementing the diet with CDC tended to increase plasma GLP-2 (P < 0.07; 39 %) and the weight of the large intestine (P < 0.10; 11 %), and increased ileal crypt depth (P < 0.04; 15 %) after 14 days of treatment exposure. Although feed intake and BW gain were not affected by treatments, feeding CDC induced the expression of the cytokines TNF-α (P < 0.02; 1.9 fold), IL-6 (P < 0.01; 2.4 fold), and IL-10 (P < 0.006; 2.2 fold) and the tight junctional protein ZON-1 (P < 0.02; 1.5 fold) in the distal small intestine. CONCLUSIONS: This study showed that the oral administration of CDC to early-weaned pigs has the potential to improve the protection of the intestinal mucosa independently of relevant changes in gut growth.

Positional Distribution of Fatty Acids in Triacylglycerols and Phospholipids from Fillets of Atlantic Salmon (Salmo Salar) Fed Vegetable and Fish Oil Blends.

The nutritional and functional characteristics of dietary fat are related to the fatty acid (FA) composition and its positional distribution in the triacylglycerol (TAG) fraction. Atlantic salmon is an important source of healthy long chain omega 3 FA (particularly, eicosapentaenoic (EPA) and docoxahexaenoic (DHA) acids). However, the impact of lipid sources in salmon feeds on the regiospecificity of FA in the fish TAG remains to be explored. The present study determines the effect of feeding salmon with blends of palm, rapeseed, and fish oil, providing two different EPA + DHA concentrations (high: H-ED 10.3% and low: L-ED 4.6%) on the fillet lipid class composition and the positional distribution of FA in TAG and phospholipids. The regiospecific analysis of fillet TAG showed that around 50% of the EPA and around 80% of DHA was located in the sn-2 position. The positional distribution of FA in phosphatidylcholine (PC), showed that around 80% of the EPA and around 90% of DHA were located in the sn-2. Fish fed the vegetable-rich diets showed higher EPA in the sn-2 position in PC (77% vs. 83% in the H-ED and L-ED diets, respectively) but similar DHA concentrations. It is concluded that feeding salmon with different EPA + DHA concentrations does not affect their positional distribution in the fillet TAG.

Atlantic salmon (Salmo salar L.) as a marine functional source of gamma-tocopherol.

Gamma tocopherol (gT) exhibits beneficial cardiovascular effects partly due to its anti-inflammatory activity. Important sources of gT are vegetable oils. However, little is known to what extent gT can be transferred into marine animal species such as Atlantic salmon by feeding. Therefore, in this study we have investigated the transfer of dietary gT into salmon. To this end, fish were fed a diet supplemented with 170 ppm gT for 16 weeks whereby alpha tocopherol levels were adjusted to 190 ppm in this and the control diet. Feeding gT-rich diets resulted in a three-fold increase in gT concentrations in the liver and fillet compared to non-gT-supplemented controls. Tissue alpha tocopherol levels were not decreased indicating no antagonistic interaction between gamma- and alpha tocopherol in salmon. The concentration of total omega 3 fatty acids slightly increased in response to dietary gT. Furthermore, dietary gT significantly decreased malondialdehyde in the fillet, determined as a biomarker of lipid peroxidation. In the liver of gT fed salmon we observed an overall down-regulation of genes involved in lipid homeostasis. Additionally, gT improved the antioxidant capacity by up-regulating Gpx4a gene expression in the pyloric caeca. We suggest that Atlantic salmon may provide a marine functional source capable of enriching gT for human consumption.

Non-protein nitrogen supplementation on in vitro fermentation profile, methane production, and microbial nitrogen synthesis in a corn silage-based substrate.

Non-protein nitrogen (NPN) supplements improve animal performance in backgrounding diets. However, there is scarce information regarding the effect of different NPN sources and combinations on ruminal fermentation profile. The current study aimed to evaluate the effect of different NPN sources and their combinations on in vitro fermentation, microbial N synthesis, and methane (CH4) production in a backgrounding diet. Incubations were conducted on three separate days for 24 h using corn silage and cotton gin byproduct (70% and 30% of DM, respectively) as substrate. Treatments were control (without NPN), urea, and five different proportions of urea-biuret and nitrate (100:0, 75:25, 50:50, 25:75, and 0:100). Each treatment, except control, was formulated to be isonitrogenous and equivalent to 1% urea inclusion. Ruminal fluid was collected from two ruminally cannulated Angus crossbred steers fed ad libitum corn silage and cotton gin byproduct plus 100 g of a urea-biuret-nitrate mixture. The concentration of volatile fatty acids (VFAs) and ammonia nitrogen (NH3-N) were determined at 12 and 24 h of incubation. Final pH, in vitro dry and organic matter digestibility, total gas production, and concentration of CH4 were determined at 24 h. The supplementation of NPN increased (P < 0.05) the concentration of NH3-N at 12 and 24 h. Although NPN supplementation increased (P < 0.05) the concentration of total VFA and acetate at 12 h, treatments did not differ (P > 0.05) at 24 h. Supplementation of NPN increased (P < 0.05) the proportion of acetate at 12 and 24 h but tended to reduce (P = 0.054) the proportion of propionate only at 12 h. Digestibility and pH were not different (P > 0.05) among treatments. Increasing nitrates in the NPN supplement increased (P < 0.05) the proportion of acetate and reduced (P < 0.05) the proportion of butyrate at 12 and 24 h. The supplementation of NPN increased (P < 0.05) microbial N synthesis. Furthermore, increasing nitrate proportion in the NPN supplement increased (P < 0.05) the microbial N synthesis and efficiency of N use. Supplementation of NPN did not modify (P > 0.05) total gas or CH4 production. However, increasing nitrate proportion in the NPN supplement linearly reduced (P < 0.05) CH4 production. Supplementation of NPN increased NH3-N concentration and microbial N while increasing the inclusion of nitrate decreased the production of CH4 and increased the microbial N synthesis in a corn silage-based substrate under in vitro conditions.

Impact of Supplementing a Backgrounding Diet with Nonprotein Nitrogen on In Vitro Methane Production, Nutrient Digestibility, and Steer Performance.

Two experiments were conducted to evaluate the effect of nonprotein nitrogen (NPN) supplementation on in vitro fermentation and animal performance using a backgrounding diet. In experiment 1, incubations were conducted on three separate days (replicates). Treatments were control (CTL, without NPN), urea (U), urea-biuret (UB), and urea-biuret-nitrate (UBN) mixtures. Except for control, treatments were isonitrogenous using 1% U inclusion as a reference. Ruminal fluid was collected from two Angus-crossbred steers fed a backgrounding diet plus 100 g of a UBN mixture for at least 35 d. The concentration of volatile fatty acids (VFA) and ammonia nitrogen (NH3-N), in vitro organic matter digestibility (IVOMD), and total gas and methane (CH4) production were determined at 24 h of incubation. In experiment 2, 72 Angus-crossbred yearling steers (303 ±â€…29 kg of body weight [BW]) were stratified by BW and randomly allocated in nine pens (eight animals/pen and three pens/treatment). Steers consumed a backgrounding diet formulated to match the diet used in the in vitro fermentation experiment. Treatments were U, UB, and UBN and were isonitrogenous using 1% U inclusion as a reference. Steers were adapted to the NPN supplementation for 17 d. Then, digestibility evaluation was performed after 13 d of full NPN supplementation for 4 d using 36 steers (12 steers/treatment). After that, steer performance was evaluated for 56 d (24 steers/treatment). In experiment 1, NPN supplementation increased the concentration of NH3-N and VFA (P < 0.01) without affecting the IVOMD (P = 0.48), total gas (P = 0.51), and CH4 production (P = 0.57). Additionally, in vitro fermentation parameters did not differ (P > 0.05) among NPN sources. In experiment 2, NPN supplementation did not change dry matter and nutrient intake (P > 0.05). However, UB and UBN showed lower (P < 0.05) nutrient digestibility than U, except for starch (P = 0.20). Dry matter intake (P = 0.28), average daily gain (P = 0.88), and gain:feed (P = 0.63) did not differ among steers receiving NPN mixtures. In conclusion, tested NPN mixtures have the potential to be included in the backgrounding diets without any apparent negative effects on animal performance and warrant further studies to evaluate other variables to fully assess the response of feeding these novel NPN mixtures.

Nonprotein nitrogen (NPN) supplements can be used as a nitrogen source for ruminants fed low-protein diets. The most common NPN source is urea, included typically at a range between 0.5% and 1% of the diet dry matter in growing beef cattle. Although other NPN sources and mixtures are available, there is scarce information regarding their use in ruminant production. Two experiments were conducted to evaluate the effect of NPN sources on in vitro fermentation and animal performance using a backgrounding diet. In experiment 1, three different incubations were performed for 24 h. Treatments were control (without NPN), urea (U), urea­biuret (UB), and urea­biuret­nitrate (UBN) mixtures. In experiment 2, 72 crossbred yearling steers were randomly assigned to one of the following treatments: U, UB, and UBN mixtures. Diets were formulated to contain the same nitrogen concentration in both experiments. In experiment 1, supplementation of NPN increased the in vitro fermentation, but there were no differences among NPN sources. In experiment 2, steers performed similarly among NPN sources. These findings suggest that NPN mixtures have the potential to be included in the backgrounding diets without detrimental effects. Further studies should evaluate other variables (e.g., fermentation dynamic and microbial protein supply) when using these novel mixtures.

Bile acids induce glucagon-like peptide 2 secretion with limited effects on intestinal adaptation in early weaned pigs.

Early weaning is a stressful event characterized by a transient period of intestinal atrophy that may be mediated by reduced secretion of glucagon-like peptide (GLP) 2. We tested whether enterally fed bile acids or plant sterols could increase nutrient-dependent GLP-2 secretion and improve intestinal adaptation in weanling pigs. During the first 6 d after weaning, piglets were intragastrically infused once daily with either deionized water (control), chenodeoxycholic acid (CDC; 60 mg/kg body weight), or ß-sitoesterol (BSE; 100 mg/kg body weight). Infusing CDC increased plasma GLP-2 (P < 0.05) but did not affect plasma GLP-1 and feed intake. The intestinal expression of glucagon-like peptide 2 receptor, sodium-dependent bile acid transporter, farnesoid X receptor, and guanosine protein-coupled bile acid receptor genes were not affected by CDC treatment. The intragastric administration of CDC did not alter the weight and length of the intestine, yet increased the activation of caspase-3 in ileal villi (P < 0.02) and the expression of interleukin 6 (P < 0.002) in the jejunum. In contrast, infusing BSE did not affect any of the variables that were measured. Our results show that the enteral administration of the bile acid CDC potentiates the nutrient-induced secretion of endogenous GLP-2 in early-weaned pigs. Bile acid-enhanced release of GLP-2, however, did not result in improved intestinal growth, morphology, or inflammation during the postweaning degenerative phase.

Effects of Aspergillus oryzae prebiotic on dietary energy and nutrient digestibility of growing pigs.

The objective of this study was to determine the effects of Aspergillus oryzae prebiotic (AOP) on nutrient digestibility in growing pigs fed high-fiber diets. Eighteen growing barrows (initial body weight = 50.6 ± 4.9 kg) were surgically equipped with a T-cannula at the distal ileum. Corn and soybean meal-based diets were formulated with fiber from cereal grain byproducts corn (distillers dried grains with solubles, DDGS), rice (rice bran, RB), or wheat (wheat middlings, WM) to meet or exceed all nutrient requirements for 50 to 75 kg growing pigs. Three additional diets were formulated to contain 0.05% AOP supplemented at the expense of corn in the DDGS diet (DDGS + AOP), RB diet (RB + AOP), and WM diet (WM + AOP). All diets contained 0.5% of titanium dioxide as an indigestible marker. Pigs were allotted randomly to a triplicated 6 × 2 Youden square design with six diets and two successive periods. Ileal digesta and fecal samples were collected for 2 d after a 21-d adaptation period, and dry matter (DM), gross energy (GE), crude protein (CP), ether extract (EE), neutral detergent fiber (NDF), and ash were analyzed to calculate apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD). Standardized ileal digestibility (SID) of amino acids (AA) was calculated by correcting AID with basal endogenous AA losses from the same set of pigs. Pigs fed the DDGS+AOP diet had greater (P < 0.05) AID of EE compared with those fed the DDGS diet. However, supplementation of AOP did not (P > 0.05) affect AID of GE, DM, CP, NDF, ash or SID of AA of any high-fiber diet. Supplementation of 0.05% AOP increased (P < 0.05) ATTD of DM, GE, CP, NDF, and ash in DDGS, RB, and WM diets. Diet digestible energy was 35 kcal/kg greater (P < 0.05) in pigs fed AOP supplemented diets compared with those fed diets without AOP. In conclusion, supplementation of AOP increased ATTD of nutrients and energy value in high-fiber diets containing DDGS, RB, or WM.

Prophylactic feeding of neomycin to Holstein calves alters gut microbiota, bile acid metabolism, and expression of genes involved in immunometabolic regulation.

The objective of this study was to evaluate the effects of prophylactic neomycin administration on Holstein bull calves' intestinal microbiota, bile acid (BA) metabolism, and transcript abundance of genes related to BA metabolism. A total of 36 calves were blocked by body weight and assigned to either non-medicated milk replacer (CTL), or neomycin for 14 days (ST) or 28 days (LT) in their milk replacer. At the end of the study, calves were euthanized to collect tissue and digesta samples from the gastrointestinal tract, liver, and adipose tissue for analysis of intestinal microbial diversity, bile acid concentration and profile in various body tissues, and gene expression related to bile acid, lipid, carbohydrate metabolism, and inflammation. Calves that received prophylactic administration of neomycin for 28 d (LT) had reduced species richness (chao1 index), and tended to have reduced phylogenetic diversity in the ileum tissue. The relative abundance of Lactobacillus, and Bifidobacterium in ileum and colon digesta were decreased in LT compared with CTL. Concentrations of primary, secondary, and total BA were increased by ST in ileal tissue. In plasma, ST and LT treatments had lower concentrations of secondary BA. Gene expression of the BA receptor FXR was increased in ileum and liver by LT compared to CTL. The expression of FXR and TGR5 in the liver was increased in the ST group compared with CTL, and in adipose tissue, 5 genes related to triglyceride, gluconeogenesis, and immune activation were differentially expressed between CTL and ST. In conclusion, we provide evidence that prophylactic administration of neomycin leads to aberrant changes in BA concentration and profile in different compartments of the enterohepatic system through a process that possibly entails antimicrobial disruption of key bacterial groups, which persists even after cessation of neomycin administration. Additionally, we uncovered an apparent link between dysregulated BA metabolism and changes in lipid metabolism and immune activation in adipose tissue and liver.

Effects of Aspergillus oryzae prebiotic on animal performance, nutrients digestibility, and feeding behavior of backgrounding beef heifers fed with either a sorghum silage- or a byproducts-based diet.

Eighty-four Angus crossbred heifers (13 ± 1 mo of age, 329.5 ± 61.92 kg of body weight [BW]) were used in a generalized randomized block design with a 2 × 2 factorial arrangement of treatments. The factors evaluated were: 1) diet type (whole plant sorghum silage [SS] vs. byproducts-based [BP]), and 2) feed additive: Aspergillus oryzae prebiotic (AOP; 2 g/d) vs. Negative control (CTL; 0 g/d), resulting in four treatments: sorghum silage-control (SC), sorghum silage-AOP (SA), byproducts-control (BC), and byproducts-AOP (BA). Heifers were stratified by body weight (BW), randomly assigned to treatments (21 heifers per treatment) and housed in 12 pens equipped with two GrowSafe feed bunks each to measure individual dry matter intake (DMI). After a 14-d adaptation, BW was measured every 14 d for 56 d. Chewing activity was monitored through collar-mounted HR-Tags (heat-related tags). Following the performance period, apparent total tract digestibility was measured in 40 heifers, using indigestible neutral detergent fiber as a marker. Heifers fed with the BP diets had greater DMI (2.92% vs. 2.59% of BW, P < 0.01) and average daily gain (ADG; 1.16 vs. 0.68 kg, P ≤ 0.01) than heifers fed with SS diets. Compared with BP-fed animals, heifers consuming the SS diets had 23 more visits/d to the feed bunks (P ≤ 0.01), consumed 53% less dry matter on each visit (P ≤ 0.01), and spent 39% more min chewing/d and 63% more min chewing/kg of DMI (P ≤ 0.01). However, chewing measured in min/kg of neutral detergent fiber intake was not affected by treatment (average 111.3 min/kg of NDF intake). Feeding AOP improved gain:feed (GF) by 15% in BP-fed heifers (0.120 vs. 0.104 kg/kg; P < 0.05). Inclusion of AOP increased organic matter digestibility (OMD) in SS diets (55.88% vs. 49.83%; P < 0.01), whereas it decreased OMD in BP diets (61.67% vs. 65.77%; P < 0.05). In conclusion, ADG and GF of BP-fed heifers was greater than SS-fed heifers, and GF was greater with AOP supplementation in BP-fed heifers. Improvement in GF in BP-fed heifers was likely not related to differences in nutrient digestibility as AOP inclusion did not enhance digestibility in the BP diet. Additionally, the effects of the AOP inclusion appear to be diet-dependent, where the 15% improvement in GF by AOP occurred in heifers fed with the more fermentable diet. Therefore, further research should explore the mechanisms responsible for the observed improvements in growth performance when feeding AOP to BP-fed heifers.

This experiment evaluated the effects of the dietary inclusion or not of Aspergillus oryzae prebiotic (AOP; 2 g/d) in two contrasting diets: sorghum silage-based (SS) vs. byproducts-based (BP), on growth performance, nutrient digestibility, and feeding behavior of growing heifers. A total of 84 Angus crossbred heifers were used in the study. Heifers fed with the BP diets had greater feed intake, average daily gain, and final body weight. In addition, heifers fed with the BP diets had reduced number of visits to the feed bunk but consumed more in each visit than heifers fed with the SS diets. Additionally, heifers fed with the BP diets had lesser chewing activity measured in total min/d and in min/kg of dry matter intake; however, chewing activity measured in min/kg of neutral detergent fiber was not influenced by treatments. The inclusion of AOP increased the gain:feed ratio by 15% in heifers fed with the BP diet but did not influence this variable in the SS diet. The inclusion of AOP increased nutrient digestibility in heifers fed with the SS diet and decreased nutrient digestibility in heifers fed with the BP diet. These results show that feeding AOP can enhance growth performance in beef heifers in a diet-dependent manner.

Daily fecal pH pattern and variation in lactating dairy cows.

Evidence supports a causal link between anomalous intestinal function and impaired performance in dairy cows. Consequently, digesta pH values obtained from colon, cecum, and rectum are increasingly used to monitor intestinal function in dairy cows. We conducted a study to describe the daily dynamics of fecal pH in lactating dairy cows. The study lasted 4 d and individual records of dry matter intake, milk yield, and fecal pH were taken. Samples of feces were taken every 4 h during the 4-d study, and sampling time was adjusted ahead by 1 h daily so that a sample was obtained for each 1-h interval of the day. Data were analyzed using a mixed-effect model including time as fixed effect and cow as a random factor. We performed a cosinor analysis using pH data at different time points to determine whether fecal pH followed a biorhythmic pattern. On average, cows consumed 19.1 ± 1.55 kg/d of dry matter and produced 26.3 ± 4.16 kg/d of milk. The most relevant results confirmed a biorhythmic pattern for feces pH around feeding time: mesor (midline estimating statistic of rhythm) 6.20, amplitude 0.28, and acrophase 5.66. Additionally, we found a positive relationship between dry matter intake and amplitude, possibly because of an increase in the amount of fermentable carbohydrate reaching the hindgut in response to increasing intake. When using fecal pH as an indicator of intestinal function, it is critical to obtain samples at several time points to capture its daily rhythmicity and to report sampling time relative to feeding.

Triterpenes from Olea europaea modulate in vitro ruminal fermentation.

Bioactive compounds present in Olea europaea have shown promising antimicrobial potential as an alternative to conventional coccidiostats. These effects are exerted by triterpenic acids (TT) present in the olive plant, namely, oleanolic acid (OA), ursolic acid (UA), and maslinic acid (MA). The objective of this study was to determine the effects of OA, UA, and MA on in vitro ruminal fermentation in comparison with monensin (MON). The study consisted of two experiments conducted as randomized complete block designs using bahiagrass hay or a high-concentrate mixed ration as basal substrates. In the first experiment (Exp. 1), a batch culture was performed with increasing doses of OA, UA, or MA. In Exp. 2, to increase the solubility of OA, two chemical forms were evaluated: a sodium salt (OA-NA) or a phyto-phospholipid complex (OA-PHYT) at 0, 4, 40, 100, and 200 mg/L of incubation inoculum. In both experiments, the dose 0 was used as control (CTL) and monensin (MON) as a positive control. Data were analyzed as a randomized complete block design with a factorial arrangement of treatments. For Exp. 2, orthogonal polynomial contrasts, adjusted for unequal spacing were used to determine the linear effects of increasing doses of OA-NA and OA-PHYT. In Exp. 1, OA reduced the concentration of CH4 in the high-concentrate substrate compared with CTL (P = 0.04). In Exp. 2 the total gas production was linearly decreased with increasing doses of OA-NA in both substrates (P ≤ 0.02). Furthermore, OA-NA and OA-PHYT decreased in vitro organic matter digestibility (P < 0.01) in the bahiagrass substrate to the same extent that MON did. However, the concentration of CH4/g of incubated DM was only reduced by the highest doses of OA-NA (P < 0.02). Lastly, no effects were observed for total VFA nor the VFA profile; however, OA-NA linearly decreased the A:P ratio in the bahiagrass substrate (P = 0.03). In conclusion, the acid form of OA as well as the sodium salt and phyto-phospholipid complex of OA were able to modify some fermentation parameters in this study; however, the magnitude of the responses was lower compared with monensin. Future studies should test OA in vivo to determine if the effects on ruminal fermentation observed here can translate into improve production efficiency while reducing carbon emissions.

Informal nutrition symposium: leveraging the microbiome (and the metabolome) for poultry production.

Knowledge of gut microbiology of poultry has advanced from a limited ability to culture relatively few microbial species, to attempting to understand the complex interactions between the bird and its microbiome. The Informal Nutrition Symposium 2021 was intended to help poultry scientists to make sense of the implications of the vast amounts of information being generated by researchers. This paper represents a compilation of the talks given at the symposium by leading international researchers in this field. The symposium began with an overview of the historical developments in the field of intestinal microbiology and microbiome research in poultry. Next, the systemic effects of the microbiome on health in the context of the interplay between the intestinal microbiota and the immune system were presented. Because the microbiome and the host communicate and influence each other, the novel field of kinomics (the study of protein phosphorylation) as used in the study of the poultry microbiome was discussed. Protein phosphorylation is a rapid response to the complex of signals among the microbiome, intestinal lumen metabolites, and the host. Then, a description of why an understanding of the role of microbial endocrinology in poultry production can lead to new understanding of the mechanisms by which the gut microbiota and the host can interact in defined mechanisms that ultimately determine health, pathogenesis of infectious disease, and behavior was given. Finally, a view forward was presented underscoring the importance of understanding mechanisms in microbiomes in other organ systems and other species. Additionally, the importance of the development of new -omics platforms and data management tools to more completely understand host microbiomes was stressed.

Avens Root (Geum Urbanum L.) Extract Discovered by Target-Based Screening Exhibits Antidiabetic Activity in the Hen's Egg Test Model and Drosophila melanogaster.

Medicinal plant extracts are becoming increasingly important as an alternative for traditional drugs against diabetes mellitus (DM). For this reason, we initialized a target-based screening of 111 root extracts from an open access plant extract library (PECKISH) by ascertaining their in-vitro inhibitory efficacy on α-glucosidase. The two most active extracts Geum urbanum L. (roseroot) and Rhodiola rosea L. (avens root) were further tested for their antidiabetic activities in terms of their impact on different regulatory key points of glucose homeostasis. To this end, various enzyme- and cell culture-based in-vitro assays were employed including the determination of sodium-dependent glucose transporter 1 (SGLT1) activity in Caco-2 monolayers by Ussing chambers and of glucose transporter 4 (GLUT4) translocation in a GFP-reporter cell line. Subsequently, the antidiabetic potential of the root extracts were further evaluated in in-vivo models, namely hen's eggs test and the fruit fly Drosophila melanogaster. Avens root extract was found to be a more potent inhibitor of the enzymes α-glucosidase and dipeptidyl peptidase-4 (DPP4) than roseroot extract. Most importantly, only avens root extract exhibited antidiabetic activity in the two in-vivo models eliciting a reduced blood glucose level in the in-ovo model and a decline of the triglyceride level in a dietary starch-induced D. melanogaster obesity model. Analyses of the polyphenolic composition of the avens root extract by HPLC revealed a high content of ellagic acid and its derivatives as well as ellagitannins such as pedunculagin, stenophyllanin, stachyurin, casuarinin and gemin A. In conclusion, avens root extract represents a promising medicinal plant that should be considered in further in-vivo studies on hyperglycemia in laboratory rodents and humans.