High-fat, high-fructose, high-cholesterol feeding causes severe NASH and cecal microbiota dysbiosis in juvenile Ossabaw swine.

Pediatric obesity and nonalcoholic steatohepatitis (NASH) are on the rise in industrialized countries, yet our ability to mechanistically examine this relationship is limited by the lack of a suitable higher animal models. Here, we examined the effects of high-fat, high-fructose corn syrup, high-cholesterol Western-style diet (WD)-induced obesity on NASH and cecal microbiota dysbiosis in juvenile Ossabaw swine. Juvenile female Ossabaw swine (5 wk old) were fed WD (43.0% fat; 17.8% high-fructose corn syrup; 2% cholesterol) or low-fat diet (CON/lean; 10.5% fat) for 16 wk ( n = 6 each) or 36 wk ( n = 4 each). WD-fed pigs developed obesity, dyslipidemia, and systemic insulin resistance compared with CON pigs. In addition, obese WD-fed pigs developed severe NASH, with hepatic steatosis, hepatocyte ballooning, inflammatory cell infiltration, and fibrosis after 16 wk, with further exacerbation of histological inflammation and fibrosis after 36 wk of WD feeding. WD feeding also resulted in robust cecal microbiota changes including increased relative abundances of families and genera in Proteobacteria ( P < 0.05) (i.e., Enterobacteriaceae, Succinivibrionaceae, and Succinivibrio) and LPS-containing Desulfovibrionaceae and Desulfovibrio and a greater ( P < 0.05) predicted microbial metabolic function for LPS biosynthesis, LPS biosynthesis proteins, and peptidoglycan synthesis compared with CON-fed pigs. Overall, juvenile Ossabaw swine fed a high-fat, high-fructose, high-cholesterol diet develop obesity and severe microbiota dysbiosis with a proinflammatory signature and a NASH phenotype directly relevant to the pediatric/adolescent and young adult population.

Evaluation of soluble corn fiber on chemical composition and nitrogen-corrected true metabolizable energy and its effects on in vitro fermentation and in vivo responses in dogs.

Dietary fermentable fiber is known to benefit intestinal health of companion animals. Soluble corn fiber (SCF) was evaluated for its chemical composition, nitrogen-corrected true ME (TMEn) content, in vitro digestion and fermentation characteristics, and in vivo effects on nutrient digestibility, fecal fermentation end products, and modulation of the fecal microbiome of dogs. Soluble corn fiber contained 78% total dietary fiber, all present as soluble dietary fiber; 56% was low molecular weight soluble fiber (did not precipitate in 95% ethanol). The SCF also contained 26% starch and 8% resistant starch and had a TMEn value of 2.6 kcal/g. Soluble corn fiber was first subjected to in vitro hydrolytic-enzymatic digestion to determine extent of digestibility and then fermented using dog fecal inoculum, with fermentative outcomes measured at 0, 3, 6, 9, and 12 h. Hydrolytic-enzymatic digestion of SCF was only 7%. In vitro fermentation showed increased (P < 0.05) concentrations of short-chain fatty acids through 12 h, with acetate, propionate, and butyrate reaching peak concentrations of 1,803, 926, and 112 µmol/g DM, respectively. Fermentability of SCF was higher (P < 0.05) than for cellulose but lower (P < 0.05) than for pectin. In the in vivo experiment, 10 female dogs (6.4 ± 0.2 yr and 22 ± 2.1 kg) received 5 diets with graded concentrations of SCF (0, 0.5, 0.75, 1.0, or 1.25% [as-is basis]) replacing cellulose in a replicated 5 × 5 Latin square design. Dogs were first acclimated to the experimental diets for 10 d followed by 4 d of total fecal collection. Fresh fecal samples were collected to measure fecal pH and fermentation end products and permit a microbiome analysis. For microbiome analysis, extraction of DNA was followed by amplification of the V4 to V6 variable region of the 16S rRNA gene using barcoded primers. Sequences were classified into taxonomic levels using a nucleotide basic local alignment search tool (BLASTn) against a curated GreenGenes database. Few changes in nutrient digestibility or fecal fermentation end products or stool consistency were observed, and no appreciable modulation of the fecal microbiome occurred. In conclusion, SCF was fermentable in vitro, but higher dietary concentrations may be necessary to elicit potential in vivo responses.

Potato fiber as a dietary fiber source in dog foods.

Potato fiber (PF), a coproduct of potato starch manufacture, was evaluated as a potential novel fiber source in dog food. Potato fiber contained 55% total dietary fiber, 29% starch, 4% crude protein, and 2% acid-hydrolyzed fat. The PF substrate was evaluated for chemical composition, in vitro digestion and fermentation characteristics, and in vivo responses. For the in vitro hydrolytic-enzymatic digestion and fermentation experiment, raw and cooked PF substrates were first subjected to hydrolytic-enzymatic digestion to determine OM disappearance and then fermented using dog fecal inoculum. Fermentation characteristics were then measured at 0, 3, 6, 9, and 12 h. For the in vivo experiment, 10 female mixed-breed dogs (6.13±0.17 yr; 22±2.1 kg) were provided 5 diets with graded concentrations (0%, 1.5%, 3%, 4.5%, or 6%) of PF in a replicated 5×5 Latin square design. Dogs were acclimated to the test diet for 10 d, followed by 4 d of total fecal collection. Fresh fecal samples were collected to measure fecal pH and fermentation end products. In vitro digestion revealed that raw and cooked PF were 32.3% and 27.9% digested enzymatically, whereas in vitro fermentation showed that PF was fermentable through 9 h. Raw PF had greater (P<0.05) acetate, propionate, and total short-chain fatty acid (SCFA) concentrations at the 12-h time point compared with cooked PF. The in vivo experiment showed no differences in apparent total tract DM, OM, CP, acid-hydrolyzed fat, or energy digestibility of diets containing graded concentrations of PF. However, total dietary fiber digestibility exhibited a linear increase (P<0.01) with increasing PF concentrations in the diet. Overall, linear increases (P<0.01) were observed for all individual and total SCFA, with a concomitant linear decrease (P<0.01) in fecal pH with increasing dietary PF. Fecal protein catabolite concentrations were low or undetectable, with the exception of spermidine, which exhibited a linear increase with increasing concentrations of PF. These findings indicated that inclusion of PF elicited favorable fermentation characteristics without negatively affecting nutrient digestibility or stool characteristics, indicating that PF could be a functional dietary fiber source in dog foods.