Fate of thymol and its monoglucosides in the gastrointestinal tract of piglets

The monoterpene thymol has been proposed as a valuable alternative to in-feed antibiotics in animal production. However, the effectiveness of the antimicrobial is comprised by its fast absorption in the upper gastrointestinal tract. In this work, two glucoconjugates, thymol a-D-glucopyranoside (T alpha G) and thymol beta-D-glucopyranoside (T beta G), were compared with free thymol for their potential to deliver higher concentrations of the active compound to the distal small intestine of supplemented piglets. Additionally, an analytical method was developed and validated for the simultaneous quantification of thymol and its glucoconjugates in different matrices. In stomach contents of pigs fed with 3333 mu mol kg(-1) thymol, T alpha G, or T beta G, total thymol concentrations amounted to 3048, 2357, and 1820 mu mol kg(-1 )dry matter, respectively. In glucoconjugate-fed pigs, over 30% of this concentration was present in the unconjugated form, suggesting partial hydrolysis in the stomach. No quantifiable levels of thymol or glucoconjugates were detected in the small intestine or cecum for any treatment, indicating that conjugation with one glucose unit did not sufficiently protect thymol from early absorption

Effects of N-acetyl-cysteine supplementation through drinking water on the glutathione redox status during the weaning transition of piglets

This study investigated the effect of N-acetyl-cysteine (NAC) supplementation through drinking water on animal performance and the glutathione (GSH) redox system in weaned piglets, particularly in relation to the immediate post-weaning feed intake. To this end, 168 piglets were weaned and either fed ad libitum or fasted the first two days, and either or not administered 200 mg/L NAC via the drinking water until d14 post-weaning. Next to animal performance until day 42 (d42), the GSH redox system was measured in erythrocytes, small intestinal mucosa, liver, lung, and kidney tissue at d0, d2, and d14 post-weaning. Animal performance and GSH levels were not affected by NAC, nor by fasting. Irrespective of treatment, a significant drop in GSH at d2 post-weaning was found as compared to d0, in particular in liver (69%), distal jejunal mucosa (72%), and lung tissue (80%). Post-weaning changes of the GSH redox status were strongly tissue-dependent. To conclude, this research indicates that GSH redox homeostasis was largely affected in multiple organs during the weaning transition. NAC supplementation did not increase GSH levels in any tissue, not even in fasted animals, questioning the fact if cysteine is the first or only limiting factor determining the rate of GSH synthesis in the early post-weaning phase

Weaning affects the glycosidase activity towards phenolic glycosides in the gut of piglets

Phenolic compounds in pig diets, originating either from feed ingredients or additives, may occur as glycosides, that is conjugated to sugar moieties. Upon ingestion, their bioavailability and functionality depend on hydrolysis of the glycosidic bond by endogenous or microbial glycosidases. Hence, it is essential to map the glycosidase activities towards phenolic glycosides present along gut. Therefore, the activity of three key glycosidases, that is alpha-glucosidase (alpha GLU), beta-glucosidase (beta GLU) and beta-galactosidase (beta GAL), was quantified in small intestinal mucosa and digesta of piglets at different gastrointestinal sites (stomach, three parts of small intestine, caecum and colon) and at different ages around weaning (10 days before and 0, 2, 5, 14 and 28 days after weaning). Activity assays were performed with p-nitrophenyl glycosides at neutral pH. The alpha GLU activities in mucosa and digesta were low (overall means 1.4 and 60 U respectively) as compared to beta GLU (15.2 and 199 U) and beta GAL (23.4 and 298 U; p < .001). Moreover, alpha GLU activity in mucosa was unaffected by age. Conversely, beta GLU and beta GAL activities dropped significantly after weaning. Minimal levels, ranging between 18% and 54% of the pre-weaning values, were reached at 5 days post-weaning. Similarly, in small intestinal digesta, reductions from 60% up to 90% were observed for the three enzyme activities on day five post-weaning as compared to pre-weaning levels. In caecal contents, activities were lowest at 14 days post-weaning, while in stomach and colon no clear weaning-induced effects were observed. Our data suggest that weaning affects the glycosidase activity in mucosa (mainly endogenous origin) and digesta (primarily bacterial origin) with the most pronounced effects occurring 5 days post-weaning. Moreover, differences in activities exist between different glycosidases and between gut locations. These insights can facilitate the prediction of the fate of existing and newly synthetized glycosides after oral ingestion in piglets

Effects of thymol and thymol α-D-Glucopyranoside on intestinal function and microbiota of weaned pigs

The present study evaluated gluco-conjugation as a measure to delay thymol absorption and enhance its antimicrobial activity in the gut of weaned piglets. The three dietary treatments consisted of a basal diet without additives (TCON), supplemented with thymol at 3.7 mmol/kg dry matter (TTHY), or with an equimolar amount of thymol α-D-glucopyranoside (TTαG). Each dietary treatment was replicated in 6 pens with 2 piglets per pen (n = 12 for analytical parameters) and was supplemented for 14 days. The total (free plus gluco-conjugated) thymol concentrations in the stomach contents were 14% lower in TTαG as compared to TTHY piglets. Neither of the additives could be detected further down the gut. E.coli counts in the proximal small intestine were significantly lower in TTHY than in TTαG pigs (3.35 vs. 4.29 log10 CFU/g); however, other bacterial counts and their metabolites were unaffected by treatment. A metagenomic bacterial analysis revealed a great relative abundance of Lactobacillus spp. in the distal small intestine (range 88.4%–99.9%), irrespective of treatment. The intestinal barrier function was improved by TTHY, but not TTαG, compared to TCON.In conclusion, gluco-conjugation did not result in higher thymol concentrations in the gut, but conversely, it seemed to diminish the biological effects of thymol in vivo

The effect of dietary quercetin on the glutathione redox system and small intestinal functionality of weaned piglets

Quercetin has been shown to alleviate mucosal damage and modulate the glutathione (GSH) redox system in the colon of rodents. In the current study, we assessed whether quercetin was able to mitigate small intestinal dysfunction in weaned pigs. Here, 224 weaned piglets were fed a diet containing quercetin at either 0, 100, 300, or 900 mg/kg diet until d14 post-weaning, followed by a common basal diet until d42. Eight animals per treatment were sampled at d5 and d14 post-weaning. In these animals, the small intestinal histomorphology, barrier function, and protein abundance of occludin, caspase-3, and proliferating cell nuclear antigen were assessed. None of these parameters were affected, and neither did quercetin improve performance up to d42 post-weaning. The GSH redox system was evaluated in blood, small intestinal mucosa, and liver. Quercetin did not affect the glutathione peroxidase, glutathione reductase, and glutamate–cysteine ligase activity in these tissues. In contrast, the hepatic glutathione transferase (GST) activity was significantly increased by quercetin supplementation at d5 post-weaning of 100, 300, and 900 mg/kg. Importantly, d5 was characterized by a more oxidized GSH redox status. To conclude, dietary quercetin had little effect on the small intestine, but did upregulate hepatic GST in the occurrence of redox disturbance

Expression of transient receptor potential ankyrin 1 and transient receptor potential vanilloid 1 in the gut of the peri-weaning pig is strongly dependent on age and intestinal site

Transient receptor potential (TRP) channels contribute to sensory transduction in the body, agonized by a variety of stimuli, such as phytochemicals, and they are predominantly distributed in afferent neurons. Evidence indicates their expression in non-neuronal cells, demonstrating their ability to modulate gastrointestinal function. Targeting TRP channels could potentially be used to regulate gastrointestinal secretion and motility, yet their expression in the pig is unknown. This study investigated TRPA1 and TRPV1 expression in different gut locations of piglets of varying age. Colocalization with enteroendocrine cells was established by immunohistochemistry. Both channels were expressed in the gut mucosa. TRPV1 mRNA abundance increased gradually in the stomach and small intestine with age, most notably in the distal small intestine. In contrast, TRPA1 exhibited sustained expression across ages and locations, with the exception of higher expression in the pylorus at weaning. Immunohistochemistry confirmed the endocrine nature of both channels, showing the highest frequency of colocalization in enteroendocrine cells for TRPA1. Specific co-localization on GLP-1 immunoreactive cells indicated their possible role in GLP-1 release and the concomitant intestinal feedback mechanism. Our results indicate that TRPA1 and TRPV1 could play a role in gut enteroendocrine activity. Moreover, age and location in the gut significantly affected gene expression

Effect of a carotenoid-producing Bacillus strain on intestinal barrier integrity and systemic delivery of carotenoids : a randomised trial in animals and humans

The aim of the present study was to investigate effects of the carotenoid-producing Bacillus indicus strain PD01 on intestinal barrier function and its ability to survive passage through the gastrointestinal tract and to assess systemic bioavailability of these carotenoids in vivo. As model for impaired barrier function, 16 early weaned piglets were randomly assigned to a control diet or control diet with PD01 for 23 days. In addition, 67 overweight/obese, otherwise healthy individuals were randomly assigned to groups receiving PD01 or placebo for 6 weeks. PD01 survived passage through the gastrointestinal tract in piglets and human subjects and resulted in significant accumulation of PD01 derived carotenoids (methyl-glycosyl-apo-8'-lycopenoate and glycosyl-apo-8'- lycopene) in human plasma after 3- and 6-weeks supplementation versus baseline (0.044 and 0.076 vs 0 mu M, respectively; p = 0.104). In summary, PD01 survived transit through the gastrointestinal tract, resulted in systemic carotenoid accumulation and improved compromised barrier function outcomes

Antibiotic affects the gut microbiota composition and expression of genes related to lipid metabolism and myofiber types in skeletal muscle of piglets

Background: Early-life antibiotic administration is known to affect gut microbiota and host adiposity, but the effects of antibiotic exposure on skeletal muscle properties remain unknown. The present study evaluated the changes in skeletal muscle properties including myofiber characteristics and composition, as well as intramuscular fat (IMF) content in skeletal muscle of piglets when exposed to a tylosin-containing diet. Results: A total of 18 piglets (28 days of age) were randomly allocated into two groups: control basal diet (Control) and Control + 100 mg tylosin phosphate/kg of feed (Antibiotic). The trial lasted for 39 days. High-throughput amplicon sequencing revealed that no significant difference in initial gut microbiota composition was existed between Control and Antibiotic groups. Antibiotic administration increased body weight and growth rate and decreased feed to gain ratio of pigs (P < 0.05). The carcass lean and fat volumes of pigs were increased by the tylosin administration (P < 0.05). Antibiotic treatment increased myofiber density and the expression of genes related to type I and type IIb myofibers in longissimus muscle (P < 0.05). The IMF content in longissimus muscle was increased by antibiotic exposure (P < 0.05). Antibiotic administration increased expression of genes related to fatty acid uptake and de novo synthesis, and decreased expression of genes related to triglyceride hydrolysis (P < 0.05). Tylosin administration affected taxonomic distribution and beta diversity of the caecal and colonic microbiota of piglets. Conclusion: These results confirm that the growth performance, myofiber composition and muscle lipid metabolism are affected by antibiotic administration, which may be associated with an altered gut microbiota, suggesting that the gut microbiota could be served as a potential target for modulating skeletal muscle properties of host