Resistant starch induces catabolic but suppresses immune and cell division pathways and changes the microbiome in the proximal colon of male pigs.

Consumption of resistant starch (RS) has been associated with various intestinal health benefits, but knowledge of its effects on global gene expression in the colon is limited. The main objective of the current study was to identify genes affected by RS in the proximal colon to infer which biologic pathways were modulated. Ten 17-wk-old male pigs, fitted with a cannula in the proximal colon for repeated collection of tissue biopsy samples and luminal content, were fed a digestible starch (DS) diet or a diet high in RS (34%) for 2 consecutive periods of 14 d in a crossover design. Analysis of the colonic transcriptome profiles revealed that, upon RS feeding, oxidative metabolic pathways, such as the tricarboxylic acid cycle and ß-oxidation, were induced, whereas many immune response pathways, including adaptive and innate immune system, as well as cell division were suppressed. The nuclear receptor peroxisome proliferator-activated receptor γ was identified as a potential key upstream regulator. RS significantly (P < 0.05) increased the relative abundance of several butyrate-producing microbial groups, including the butyrate producers Faecalibacterium prausnitzii and Megasphaera elsdenii, and reduced the abundance of potentially pathogenic members of the genus Leptospira and the phylum Proteobacteria. Concentrations in carotid plasma of the 3 main short-chain fatty acids acetate, propionate, and butyrate were significantly higher with RS consumption compared with DS consumption. Overall, this study provides novel insights on effects of RS in proximal colon and contributes to our understanding of a healthy diet.

A diet high in resistant starch modulates microbiota composition, SCFA concentrations, and gene expression in pig intestine.

Resistant starch (RS) is highly fermentable by microbiota in the colon, resulting in the production of SCFAs. RS is thought to mediate a large proportion of its health benefits, including increased satiety, through the actions of SCFAs. The aim of this study was to investigate the effects of a diet high in RS on luminal microbiota composition, luminal SCFA concentrations, and the expression of host genes involved in SCFA uptake, SCFA signaling, and satiety regulation in mucosal tissue obtained from small intestine, cecum, and colon. Twenty adult female pigs were either assigned to a digestible starch (DS) diet or a diet high in RS (34%) for a period of 2 wk. After the intervention, luminal content and mucosal scrapings were obtained for detailed molecular analysis. RS was completely degraded in the cecum. In both the cecum and colon, differences in microbiota composition were observed between DS- and RS-fed pigs. In the colon these included the stimulation of the healthy gut-associated butyrate-producing Faecalibacterium prausnitzii, whereas potentially pathogenic members of the Gammaproteobacteria, including Escherichia coli and Pseudomonas spp., were reduced in relative abundance. Cecal and colonic SCFA concentrations were significantly greater in RS-fed pigs, and cecal gene expression of monocarboxylate transporter 1 (SLC16A1) and glucagon (GCG) was induced by RS. In conclusion, our data show that RS modulates microbiota composition, SCFA concentrations, and host gene expression in pig intestine. Combined, our data provide an enhanced understanding of the interaction between diet, microbiota, and host.

Effects of hatching system on the welfare of broiler chickens in early and later life.

Broiler chicks usually hatch in the hatchery without access to feed and water until placement at the farm. This can affect their health and welfare negatively. Therefore, alternative strategies have been developed, for instance providing chicks with early nutrition in the hatchery or hatching eggs directly on-farm. However, information on the physical and mental welfare of chicks hatched in these systems compared to conventionally hatched chicks is limited. The aim of this study was to investigate the effects of alternative hatching systems on the welfare of broiler chickens in early and later life. A system comparison was performed with chickens that hatched conventionally in a hatchery (HH), in a system which provided light, feed, and water in a hatcher (hatchery-fed, HF), or on-farm (on-farm hatched, OH, where feed and water were available and transport of day-old chicks from the hatchery to the farm was not necessary). Chickens were reared in 3 batches, in 12 floor pens per batch (approximately 1,155 animals per pen), with a total of 12 replicates per treatment. Animal-based welfare indicators were assessed following standard protocols: plumage cleanliness, footpad dermatitis (FPD), hock burn, skin lesions (all at day 21 and 35 of age), and gait score (day 35). Furthermore, a set of behavioral tests was carried out: novel environment (day 1 and 21), tonic immobility, novel object, and avoidance distance test (day 4 and 35). Plumage cleanliness, hock burn, and skin lesions were affected by age but not by hatching system, with older broilers scoring worse than younger ones (P < 0.05). An effect of hatching system was only found for FPD, with the highest prevalence in HH chickens, followed by HF and OH chickens (P < 0.05). All responses measured in the behavioral tests were affected by age but not by hatching system. In later life, chickens were significantly less fearful than during the first days of life. The results indicate that conventionally hatched chickens scored significantly worse for FPD, whereas, in general, hatching system seemed to have minor effects on other broiler welfare indicators.

Day-old chicken quality and performance of broiler chickens from 3 different hatching systems.

In on-farm hatching systems, eggs are transported at d 18 of incubation to the broiler farm, where chickens have immediate access to feed and water after hatching. In hatchery-fed systems, newly hatched chickens have immediate access to feed and water in the hatchery and are transported to the farm thereafter. Conventionally hatched chickens can remain without access to feed and water up to 72 h after hatching until placement on the farm. The current study compared day-old chicken quality, performance, and slaughter yield of broiler chickens that were on-farm hatched (OH), hatchery-fed (HF), or conventionally hatchery-hatched (HH). The experiment was performed in 6 rooms in 1 house. Each room contained 2 duplicate pens with approximately 1,155 chickens per pen; 2 rooms with each 2 duplicate pens were assigned to 1 treatment. The experiment was repeated during 3 consecutive production cycles. Chickens originated from young parent stock flocks. Results showed that HF and OH chickens were heavier and longer than HH chickens at day (D) 1. Relative weight of stomach and intestines were highest for OH chickens. The OH chickens had worse day-old chicken quality in terms of navel condition and red hocks than HH and HF chickens. Treatments did not differ in first wk and total mortality. From D0 until slaughter age, body weight was highest for OH, followed by HF and HH. Furthermore, carcass weight at slaughter age (D40) was highest for OH chickens, followed by HF and HH chickens. Breast fillets showed a higher incidence of white striping and wooden breast in HF and OH chickens compared with HH chickens. In conclusion, the current study showed that both OH and HF chickens of young parent flocks had better growth performance, which could explain the higher prevalence of breast myopathies, compared with HH. The worse day-old chicken quality for OH compared with HH and HF does not seem to affect first wk mortality and later life performance.

In vivo degradation of alginate in the presence and in the absence of resistant starch.

This study evaluated the intestinal degradability of alginate during 74 days intake in pigs as models for humans. Diets contained pregelatinized starch, retrograded starch, alginate, or a mix of retrograded starch and alginate. Faeces were collected on day 1, 3, 7, 14, 39 and 74. Clear trends in intestinal alginate degradation were observed. Up to day 39, the total tract digestibility of alginate was limited (0.52 ± 0.10), and was lower with the inclusion of retrograded starch in the diet (0.34 ± 0.02). More than 90% of the faecal alginate was insoluble in water, which may explain the low digestibility of the alginate. The digestibility of mannuronic acid (M) was 2-3 times higher than that of guluronic acid (G). The changes of G:M ratio and the relative amounts of alginate oligosaccharides between day 39 and 74 indicated that the microbiota needed more than 39 days to adapt to alginate. This study demonstrated that in-depth analyses of dietary fibres are valuable in understanding the fate of the dietary fibres in the large intestine as it was shown that degradation of a dietary fibre depends not only on the properties of the fibre itself, but also on the other dietary fibres present in the diet and the adaptation time.

Influence of a diet rich in resistant starch on the degradation of non-starch polysaccharides in the large intestine of pigs.

To investigate the effect of resistant starch to the degradation of other non-starch polysaccharides (NSPs) in the large intestine of pigs, two groups of pigs were fed either a diet containing digestible starch (DS) or a diet containing resistant starch (RS). Both diets contained NSPs from wheat and barley. Digesta from different parts of the large intestine were collected and analysed for sugar composition and carbohydrate-degrading-enzyme activities. Resistant starch, as well as ß-glucans and soluble arabinoxylan, was utilised mainly in the caecum. The utilisation of ß-glucans and soluble arabinoxylan in the caecum was higher in DS-fed pigs than in RS-fed pigs. Analyses on carbohydrate-degrading-enzyme activities demonstrated that microbial enzyme production was stimulated according to the diet composition, and the enzyme profile throughout the large intestine of RS-fed pigs indicated that the presence of resistant starch shifted the utilisation of NSPs to more distal parts of the colon.

Effects of dietary fibers with different fermentation characteristics on feeding motivation in adult female pigs.

Dietary fibers can be fermented in the colon, resulting in production of short-chain fatty acids (SCFA) and secretion of satiety-related peptides. Fermentation characteristics (fermentation kinetics and SCFA-profile) differ between fibers and could impact their satiating potential. We investigated the effects of fibers with varying fermentation characteristics on feeding motivation in adult female pigs. Sixteen pair-housed pigs received four diets in four periods in a Latin square design. Starch from a control (C) diet was exchanged, based on gross energy, for inulin (INU), guar gum (GG), or retrograded tapioca starch (RS), each at a low (L) and a high (H) inclusion level. This resulted in a decreased metabolizable energy intake when feeding fiber diets as compared with the C diet. According to in vitro fermentation measurements, INU is rapidly fermentable and yields relatively high amounts of propionate, GG is moderately rapidly fermentable and yields relatively high amounts of acetate, and RS is slowly fermentable and yields relatively high amounts of butyrate. Feeding motivation was assessed using behavioral tests at 1h, 3h and 7h after the morning meal, and home pen behavioral observations throughout the day. The number of wheel turns paid for a food reward in an operant test was unaffected by diet. Pigs on H-diets ran 25% slower for a food reward in a runway test than pigs on L-diets, and showed less spontaneous physical activity and less stereotypic behavior in the hours before the afternoon meal, reflecting increased interprandial satiety. Reduced feeding motivation with increasing inclusion level was most pronounced for RS, as pigs decreased speed in the runway test and tended to have a lower voluntary food intake in an ad libitum food intake test when fed RS-H. In conclusion, increasing levels of fermentable fibers in the diet seemed to enhance satiety in adult pigs, despite a reduction in metabolizable energy supply. RS was the most satiating fiber, possibly due to its slow rate of fermentation and high production of butyrate.