Effect of reducing the ratio of omega-6-to-omega-3 fatty acids in diets of low protein quality on nursery pig growth performance and immune response.

A total of 240 newly weaned pigs (5.25 ± 0.15 kg BW) were used to determine the dietary omega-6-to-omega-3 (ω-6:ω-3) fatty acid ratio that optimized growth performance and immune responses when fed corn and soybean meal (SBM)-based diets with low protein quality. Pigs were randomly assigned to 1 of 5 dietary treatments (n = 6 pens per treatment; day 0 of study): [1] positive control (High; included animal proteins and 5% corn oil), [2] negative control (Low0; corn- and SBM-based and 5% corn oil), or 1 of 3 Low diets with increasing supplementation of fish oil to replace corn oil: [3] 1.25% (Low1.25), [4] 2.5% (Low2.5), [5] 5% (Low5) to achieve 5:1, 3:1, and 1:1 ω-6:ω-3 ratios, respectively. Pigs were fed dietary treatments in 2 phases for 7 and 14 d, respectively, followed by a common phase III diet for 21 d. On day 6 and 20, 12 pigs per treatment were immune sensitized with 0.5 mg ovalbumin (OVA) and 0.5 mg Quil A adjuvant in 1 mL saline. The dermal hypersensitivity response (DHR) was evaluated on day 40 in these same pigs, using intradermal injection of OVA; changes in skin-fold thickness were measured. On day 21, 4 pigs per pen were immune challenged with LPS (30 µg Escherichia coli LPS per kg BW) or saline (n = 12); rectal temperature was monitored over 3 h. During phase I only, ADG, ADFI, and G:F were greater for pigs fed the High diet vs. those fed the Low diet (P < 0.05), and increased with increasing fish oil supplementation up to 2.5% (Low2.5), but decreased for pigs fed the Low5 diet (quadratic; P < 0.05, P = 0.086, and P < 0.05 for ADG, ADFI, and G:F, respectively). On day 21, LPS increased rectal temperature (vs. saline at 1-, 2-, and 3-h post-challenge; P < 0.001); fish oil supplementation reduced rectal temperature 2-h post-challenge in the Low-fed pigs (linear; P < 0.05). On day 22, serum haptoglobin was greatest for pigs fed Low0 and decreased with increasing fish oil supplementation (linear; P < 0.05). Immunization with OVA induced a serum anti-OVA IgG response, which was reduced on day 34 among pigs fed Low diets with increasing fish oil supplementation (linear; P = 0.050). On day 40, and 6 h after intradermal injection of OVA, the DHR was least for pigs fed the Low2.5 diet (P < 0.05). Inclusion of 2.5% fish oil (3:1, ω-6:ω-3) optimized growth performance during the early nursery phase when pigs were most sensitive to diets with low protein quality; the ideal ω-6-to-ω-3 fatty acid ratio may differ when using immune responses as the major outcome.

Fiber supplementation influences phylogenetic structure and functional capacity of the human intestinal microbiome: follow-up of a randomized controlled trial.

BACKGROUND: In our published randomized, double-blind, placebo-controlled, 3-period crossover trial, healthy adult men (n = 21) consumed bars containing no supplemental fiber (placebo; NFC), polydextrose (21 g/d), and soluble corn fiber (SCF; 21 g/d) for 21 d each. Fecal specimens were collected between days 16 and 21 for fermentative end-product analysis and 16S ribosomal RNA bacterial gene amplification for bacterial taxa identification. Fiber supplementation decreased fecal putrefaction compounds and shifted abundances of several bacterial taxa. OBJECTIVE: The objective was to perform whole-genome shotgun 454 pyrosequencing on the same fecal specimens collected in that clinical trial to obtain comprehensive fecal bacterial genome sequencing coverage and explore the full range of bacterial genetic information in the fecal microbiome, thereby using a systematic approach to study the impact of dietary fiber supplementation on fecal metabolites, bacterial taxa, and bacterial metagenomes. DESIGN: Fecal samples were subjected to whole-genome shotgun 454 pyrosequencing to identify both fecal bacterial populations present and their functional genetic capacity. RESULTS: Whole-genome shotgun sequencing results revealed that fiber consumption shifted the Bacteroidetes:Firmicutes ratio, increasing the relative abundance of Bacteroidetes 12 ± 2% and 13 ± 2% with polydextrose and SCF, respectively, compared with NFC. Bivariate correlations showed a positive correlation between the Bacteroidetes:Firmicutes ratio and total dietary fiber intake but not body mass index. Principal coordinates analysis of Bray-Curtis distances indicated that bacterial gene composition was more similar in participants consuming fibers (polydextrose and SCF combined) in comparison with NFC. Shifts in bacterial gene abundances after polydextrose and SCF supplementation included genes associated with carbohydrate, amino acid, and lipid metabolism, as well as metabolism of cofactors and vitamins. CONCLUSION: This study conveys novel information about the impact of dietary fiber supplementation on the phylogenetic structure and functional capacity of the fecal microbiome of healthy adults.

Alterations in ileal mucosa bacteria related to diet complexity and growth performance in young pigs.

BACKGROUND: Evaluation of the prolonged impact of weaning diet on ileal mucosa bacteria and during periods of reduced and improved growth was conducted using 454 pyrosequencing. METHODOLOGY/PRINCIPAL FINDINGS: Weaned pigs were fed HIGH or LOW complexity diets, with or without antibiotics, for 6 weeks, followed by a common grower diet. Pigs were killed at 2 (n = 4 or 5) and 8 (n = 6) weeks post-weaning (periods of reduced and improved growth, respectively). Mucosal bacteria were removed; DNA was extracted and amplified using the V1-V3 region of the 16S rRNA gene. Mucosal bacteria clustered more closely by week post-weaning than diet but 44% of bacterial species did not change from week 2 to 8. There was no effect of diet complexity or antibiotic inclusion on indices of bacterial diversity. Firmicutes made up 91 and 96% of total reads at week 2 and 8, respectively. The proportion of Clostridium paraputrificum increased (P = 0.003) from week 2 to 8 in pigs fed LOW but didn't change in pigs fed HIGH; whereas Clostridium leptum decreased (P = 0.02) from week 2 to 8 in pigs fed LOW but didn't change in pigs fed HIGH. The proportion of Sarcina genus was 3-fold higher in pigs fed A+ compared to A- at week 2 and 5-fold higher at week 8 despite the lack of in-feed antibiotics at that time. CONCLUSIONS/SIGNIFICANCE: Shifts in mucosal bacteria populations may be related to dietary induced changes in growth performance during reduced and improved growth but further studies are required to confirm causative relationship. Weaning diet results in species specific prolonged alterations in mucosal bacteria, particularly where high levels of in-feed antibiotics are used. A considerable portion of ileal mucosal bacteria colonize early and remain stable over time despite changes in diet.

The gut microbiome of kittens is affected by dietary protein:carbohydrate ratio and associated with blood metabolite and hormone concentrations.

High-protein, low-carbohydrate (HPLC) diets are common in cats, but their effect on the gut microbiome has been ignored. The present study was conducted to test the effects of dietary protein:carbohydrate ratio on the gut microbiota of growing kittens. Male domestic shorthair kittens were raised by mothers fed moderate-protein, moderate-carbohydrate (MPMC; n 7) or HPLC (n 7) diets, and then weaned at 8 weeks onto the same diet. Fresh faeces were collected at 8, 12 and 16 weeks; DNA was extracted, followed by amplification of the V4­V6 region of the 16S rRNA gene using 454 pyrosequencing. A total of 384 588 sequences (average of 9374 per sample) were generated. Dual hierarchical clustering indicated distinct clustering based on the protein:carbohydrate ratio regardless of age. The protein:carbohydrate ratio affected faecal bacteria. Faecal Actinobacteria were greater (P< 0·05) and Fusobacteria were lower (P< 0·05) in MPMC-fed kittens. Faecal Clostridium, Faecalibacterium, Ruminococcus, Blautia and Eubacterium were greater (P< 0·05) in HPLC-fed kittens, while Dialister, Acidaminococcus, Bifidobacterium, Megasphaera and Mitsuokella were greater (P< 0·05) in MPMC-fed kittens. Principal component analysis of faecal bacteria and blood metabolites and hormones resulted in distinct clusters. Of particular interest was the clustering of blood TAG with faecal Clostridiaceae, Eubacteriaceae, Ruminococcaceae, Fusobacteriaceae and Lachnospiraceae; blood ghrelin with faecal Coriobacteriaceae, Bifidobacteriaceae and Veillonellaceae; and blood glucose, cholesterol and leptin with faecal Lactobacillaceae. The present results demonstrate that the protein:carbohydrate ratio affects the faecal microbiome, and highlight the associations between faecal microbes and circulating hormones and metabolites that may be important in terms of satiety and host metabolism.

Feline gastrointestinal microbiota.

The close relationship between gastrointestinal (GI) microbiota and its host has an impact on the health status of an animal that reaches beyond the GI tract. A balanced microbiome stimulates the immune system, aids in the competitive exclusion of transient pathogens and provides nutritional benefits to the host. With recent rapid advances in high-throughput sequencing technology, molecular approaches have become the routinely used tools for ecological studies of the feline microbiome, and have revealed a highly diverse and complex intestinal ecosystem in the feline GI tract. The major bacterial groups are similar to those found in other mammals, with Firmicutes, Bacteroidetes, Actinobacteria and Proteobacteria constituting more than 99% of intestinal microbiota. Several nutritional studies have demonstrated that the feline microbiota can be modulated by the amount of soluble fibers (i.e., prebiotics) and macronutrients (i.e., protein content) in the diet. Initial clinical studies have suggested the presence of a dysbiosis in feline inflammatory bowel disease (IBD). Recently, metagenomic approaches have attempted to characterize the microbial gene pool. However, more studies are needed to describe the phylogenetic and functional changes in the intestinal microbiome in disease states and in response to environmental and dietary modulations. This paper reviews recent studies cataloging the microbial phylotypes in the GI tract of cats.

Current state of knowledge: the canine gastrointestinal microbiome.

Gastrointestinal (GI) microbes have important roles in the nutritional, immunological, and physiologic processes of the host. Traditional cultivation techniques have revealed bacterial density ranges from 10(4) to 10(5) colony forming units (CFU)/g in the stomach, from 10(5) to 10(7) CFU/g in the small intestine, and from 10(9) to 10(11) CFU/g in the colon of healthy dogs. As a small number of bacterial species can be grown and studied in culture, however, progress was limited until the recent emergence of DNA-based techniques. In recent years, DNA sequencing technology and bioinformatics have allowed for better phylogenetic and functional/metabolic characterization of the canine gut microbiome. Predominant phyla include Firmicutes, Bacteroidetes, Fusobacteria, Proteobacteria, and Actinobacteria. Studies using 16S ribosomal RNA (rRNA) gene pyrosequencing have demonstrated spatial differences along the GI tract and among microbes adhered to the GI mucosa compared to those in intestinal contents or feces. Similar to humans, GI microbiome dysbiosis is common in canine GI diseases such as chronic diarrhea and inflammatory bowel diseases. DNA-based assays have also identified key pathogens contributing to such conditions, including various Clostridium, Campylobacter, Salmonella, and Escherichia spp. Moreover, nutritionists have applied DNA-based techniques to study the effects of dietary interventions such as dietary fiber, prebiotics, and probiotics on the canine GI microbiome and associated health indices. Despite recent advances in the field, the canine GI microbiome is far from being fully characterized and a deeper characterization of the phylogenetic and functional/metabolic capacity of the GI microbiome in health and disease is needed. This paper provides an overview of recent studies performed to characterize the canine GI microbiome.

454 pyrosequencing reveals a shift in fecal microbiota of healthy adult men consuming polydextrose or soluble corn fiber.

The relative contribution of novel fibers such as polydextrose and soluble corn fiber (SCF) to the human gut microbiome and its association with host physiology has not been well studied. This study was conducted to test the impact of polydextrose and SCF on the composition of the human gut microbiota using 454 pyrosequencing and to identify associations among fecal microbiota and fermentative end-products. Healthy adult men (n = 20) with a mean dietary fiber (DF) intake of 14 g/d were enrolled in a randomized, double-blind, placebo-controlled crossover study. Participants consumed 3 treatment snack bars/d during each 21-d period that contained no supplemental fiber (NFC), polydextrose (PDX; 21 g/d), or SCF (21 g/d) for 21 d. There were no washout periods. Fecal samples were collected on d 16-21 of each period; DNA was extracted, followed by amplification of the V4-V6 region of the 16S rRNA gene using barcoded primers. PDX and SCF significantly affected the relative abundance of bacteria at the class, genus, and species level. The consumption of PDX and SCF led to greater fecal Clostridiaceae and Veillonellaceae and lower Eubacteriaceae compared with a NFC. The abundance of Faecalibacterium, Phascolarctobacterium, and Dialister was greater (P < 0.05) in response to PDX and SCF intake, whereas Lactobacillus was greater (P < 0.05) only after SCF intake. Faecalibacterium prausnitzii, well known for its antiinflammatory properties, was greater (P < 0.05) after fiber consumption. Principal component analysis clearly indicated a distinct clustering of individuals consuming supplemental fibers. Our data demonstrate a beneficial shift in the gut microbiome of adults consuming PDX and SCF, with potential application as prebiotics.

Effects of viscosity and fermentability of dietary fibre on nutrient digestibility and digesta characteristics in ileal-cannulated grower pigs.

Relative contributions of two functional properties, viscosity and fermentability of dietary fibre, on apparent ileal digestibility (AID), apparent total tract digestibility (ATTD), digesta passage rate, N retention and SCFA concentration have not been established. Thus, eight ileal-cannulated pigs randomised in a double 4 × 4 Latin square were fed four diets based on maize starch and casein supplemented with 5 % of actual fibre in a 2 × 2 factorial arrangement: low-fermentable, low-viscous cellulose (CEL); low-fermentable, high-viscous carboxymethylcellulose (CMC); high-fermentable, low-viscous oat ß-glucan (LBG); high-fermentable, high-viscous oat ß-glucan (HBG). Viscosity and fermentability interacted to affect (P < 0·001) digesta viscosity and AID and ATTD of nutrients. These properties tended to interact to affect (P < 0·10) digesta passage rate and butyrate. Pigs fed the CMC diet had the lowest (P < 0·05) digesta passage rate and the highest (P < 0·001) AID of energy, crude protein and DM, and ATTD of energy and DM. Post-ileal DM digestibility was highest (P < 0·001) for pigs fed the CEL and HBG diets. Post-ileal DM digestibility had a negative, curvilinear relationship with the AID of energy and crude protein (R2 0·85 and 0·72, respectively; P < 0·001). Digesta viscosity had a less strong relationship with the AID of energy and crude protein (R2 0·45 and 0·36, respectively; P < 0·001). In conclusion, high-viscous, low-fermentable dietary fibre increases the proportion of a diet that is digested in the small intestine by reducing digesta passage rate.

Dietary oat beta-glucan reduces peak net glucose flux and insulin production and modulates plasma incretin in portal-vein catheterized grower pigs.

Net glucose and SCFA flux and insulin secretion into the portal vein might be associated with the incretins glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1). Our objectives were to clarify this association and study the impact of 2 doses of dietary oat beta-glucan on the variables. Three 35-kg portal vein-catheterized pigs were fed 3 diets containing 0, 3, or 6% oat beta-glucan concentrate (BG0, BG3, and BG6) for 7 d in a repeated 3 x 3 Latin square. On d 7, blood was sampled for 12 h postprandially. Net glucose flux and apparent hormone production were calculated from plasma portal-arterial differences x flow. Postprandially, pigs fed BG6 had lower (P < 0.05) portal glucose at 15, 30, and 45 min and a lower (P < 0.05) net glucose flux during the first hour. Pigs fed BG6 tended to have lower (P < 0.10) portal C-peptide without lowering insulin, indicating that pigs fed BG6 had lower actual insulin release combined with a higher prehepatic retention of insulin. Pigs fed BG6 had lower (P < 0.05) portal GIP and GLP-1, which in turn were correlated (R(2) = 0.81 and 0.88, respectively; P < 0.01) with portal glucose. Pigs fed BG3 and BG6 had a higher (P < 0.05) net SCFA flux than pigs fed BG0, indicating increased fermentation. In conclusion, dietary supplementation of 6% oat beta-glucan concentrate decreased net glucose flux, increased net SCFA flux, and decreased peak apparent insulin production, changes that were associated with GIP and GLP-1 mediation.

Nonstarch polysaccharides modulate bacterial microbiota, pathways for butyrate production, and abundance of pathogenic Escherichia coli in the pig gastrointestinal tract.

The impact of nonstarch polysaccharides (NSP) differing in their functional properties on intestinal bacterial community composition, prevalence of butyrate production pathway genes, and occurrence of Escherichia coli virulence factors was studied for eight ileum-cannulated growing pigs by use of terminal restriction fragment length polymorphism (TRFLP) and quantitative PCR. A cornstarch- and casein-based diet was supplemented with low-viscosity, low-fermentability cellulose (CEL), with high-viscosity, low-fermentability carboxymethylcellulose (CMC), with low-viscosity, high-fermentability oat beta-glucan (LG), and with high-viscosity, high-fermentability oat beta-glucan (HG). Only minor effects of NSP fractions on the ileal bacterial community were observed, but NSP clearly changed the digestion in the small intestine. Compared to what was observed for CMC, more fermentable substrate was transferred into the large intestine with CEL, LG, and HG, resulting in higher levels of postileal dry-matter disappearance. Linear discriminant analysis of NSP and TRFLP profiles and 16S rRNA gene copy numbers for major bacterial groups revealed that CMC resulted in a distinctive bacterial community in comparison to the other NSP, which was characterized by higher gene copy numbers for total bacteria, Bacteroides-Prevotella-Porphyromonas, Clostridium cluster XIVa, and Enterobacteriaceae and increased prevalences of E. coli virulence factors in feces. The numbers of butyryl-coenzyme A (CoA) CoA transferase gene copies were higher than those of butyrate kinase gene copies in feces, and these quantities were affected by NSP. The present results suggest that the NSP fractions clearly and distinctly affected the taxonomic composition and metabolic features of the fecal microbiota. However, the effects were more linked to the individual NSP and to their effect on nutrient flow into the large intestine than to their shared functional properties.