The optimal ranges of protein for healthy adult dogs are not known. This study evaluated the impact of long-term consumption of foods containing low, medium, and high levels of protein on serum, urine, and fecal metabolites, and gut microbiome in beagles. Following maintenance on a prefeed food for 14 days, dogs (15 neutered males, 15 spayed females, aged 2–9 years, mean initial weight 11.3 kg) consumed the low (18.99%, dry matter basis), medium (25.34%), or high (45.77%) protein foods, each for 90 days, in a William’s Latin Square Design sequence. In serum and/or urine, metabolites associated with inflammation (9,10-dihydroxyoctadecanoic acid (DiHOME)), 12,13-DiHOME) and kidney dysfunction (urea, 5-hydroxyindole sulfate, 7-hydroxyindole sulfate, p-cresol sulfate) increased with higher protein levels in food, while one-carbon pathway metabolites (betaine, dimethylglycine, sarcosine) decreased. Fecal pH increased with protein consumed, and levels of beneficial indoles and short-chain fatty acids decreased while branched-chain fatty acids increased. Beta diversity of the fecal microbiome was significantly different, with increased abundances of proteolytic bacteria with higher protein food. Feeding dogs a high amount of protein leads to a shift to proteolytic gut bacteria, higher fecal pH, and is associated with increased levels of metabolites linked with inflammation and kidney dysfunction.

This study assessed changes in canine fecal metabolites and microbiota with the consumption of foods with increasing concentrations of a fiber bundle including pecan shells, flax seed, and powders of cranberry, citrus, and beet that was previously shown (at 14% w/w) to improve stool quality, shift fecal bacterial metabolism from proteolysis to saccharolysis, increase abundance of saccharolytic bacteria, and decrease abundance of proteolytic bacteria. In this study, 48 healthy adult dogs were split evenly to consume different inclusion levels (0%, 1%, 2%, and 4%) of the fiber bundle for a 31-day period following a 28-day pre-feed period. Increases from baseline in the fecal short-chain fatty acids butyric acid, valeric acid, and hexanoic acid were observed only in the dogs that consumed the food with the 4% fiber bundle. With addition of any level of the fiber bundle, increases were seen in the polyphenols hesperidin, hesperetin, ponciretin, secoisolariciresinol diglucoside, secoisolariciresinol, and enterodiol. However, fecal microbiota and their metabolism, and stool scores were largely unaffected by the fiber bundle. Overall, addition of the fiber bundle appeared to increase bioactive metabolites of increased antioxidant and anti-inflammatory potency for beneficial to health and, at levels ≥4%, shifted gut bacterial metabolism toward saccharolysis.

Consumption of fiber in its different forms can result in positive health effects. Prior studies in dogs found that addition of a fiber bundle (composed of pecan shells, flax seed, and powders of cranberry, citrus, and beet) to food resulted in a shift in fecal bacterial metabolism from proteolysis to saccharolysis. The present study evaluated the changes in fecal metabolites and microbiota in healthy cats following the consumption of this fiber bundle. Following a 28-day pre-feed period, 56 healthy adult cats received food with none or one of three concentrations (0%, 1%, 2%, and 4%) of the fiber bundle for a 31-day period. In cats that consumed the 4% fiber bundle, levels of ammonium and fecal branched-chain fatty acids (BCFAs) decreased from baseline and compared with the other groups. Addition of any level of the fiber bundle resulted in increases in beneficial metabolites: polyphenols hesperidin, hesperetin, ponciretin, secoisolariciresinol diglucoside, secoisolariciresinol, and enterodiol. Little change in fecal microbiota was observed. Since higher levels of ammonia and BCFAs indicate putrefactive metabolism, the decreases in these with the 4% fiber bundle indicate a shift toward saccharolytic metabolism despite little change in the microbiota composition.

Protein production through precision fermentation is an attractive option to reduce animal use in food production. This study focused on the safety and digestibility of food containing a test ingredient, brewed lamb protein (Saccharomyces cerevisiae expressing a lamb protein), at inclusion levels of 0% (control), 15%, 30%, or 40% were evaluated over 182 days in 40 dogs. No significant differences in body weight, body condition score, body composition, physical examination, food intake, or urinary parameters were observed among groups. Hematological and serum biochemical parameters were within reference ranges and were largely similar among groups. Levels of chloride, cholesterol, and homocysteine were significantly lower at some time points in dogs fed the test ingredient foods compared with the control food, but all remained within reference ranges. Whole blood taurine levels indicated no deficiency. Alanine aminotransferase levels were above the normal range at Day 177 in dogs in the control and 15% test ingredient groups. Fecal score was slightly decreased in dogs fed the 30% or 40% test ingredient foods, and fecal pH was decreased in dogs fed any of the test ingredient foods. Digestibility of most parameters was similar among foods, but apparent fat digestibility was significantly lower in foods with the test ingredient. Overall, these results indicate that the test ingredient is safe for use in dog food.