The effects of hydrolyzed protein on macronutrient digestibility, fecal metabolites and microbiota, oxidative stress and inflammatory biomarkers, and skin and coat quality in adult dogs.

Research on protein hydrolysates has observed various properties and functionalities on ingredients depending on the type of hydrolysate. The objective of this study was to evaluate the effects of hydrolyzed chicken protein that was incorporated into diets on digestibility, gut health, skin and coat health, oxidative stress, and intestinal inflammation markers in healthy adult dogs. Five complete and balanced diets were manufactured: (1) CONd: 25% chicken meal diet; (2) 5% CLHd: 5% chicken liver and heart hydrolysate plus 20% chicken meal diet; (3) CLHd: 25% chicken liver and heart hydrolysate diet; (4) 5% CHd: 5% chicken hydrolysate plus 20% chicken meal diet; (5) CHd: 25% chicken hydrolysate diet. A replicated 5 × 5 Latin square design was used which included 10 neutered adult Beagles. Each of the 5 periods consisted of a 7-d washout time and a 28-d treatment period. All diets were well accepted by the dogs. Fecal butyrate concentration was higher while fecal isovalerate and total phenol/indole were lower in dogs fed CLHd than CONd (P < 0.05). Dogs fed CHd had higher fecal immunoglobulin A concentration when compared with CLHd (P < 0.05); however, both groups were comparable to the CONd. There was no difference among groups in serum cytokine concentrations, serum oxidative stress biomarkers, or skin and coat health analyses (P > 0.05). Fecal microbiota was shifted by CLHd with higher abundance in Ruminococcus gauvreauii group as well as lower Clostridium sensu stricto 1, Sutterella, Fusobacterium, and Bacteroides when compared with CONd (P < 0.05). There was also a difference in beta diversity of fecal microbiota between CLHd and CHd (P < 0.05). In conclusion, chicken protein hydrolysate could be incorporated into canine extruded diets as a comparable source of protein to traditional chicken meal. The test chicken protein hydrolysates showed the potential to support gut health by modulating immune response and microbiota; however, functional properties of protein hydrolysates are dependent on inclusion level and source.

Research has been exploring various functional properties of hydrolyzed protein to benefit the health of animals. However, the functionality of a hydrolyzed protein depends on the type of hydrolysate. Therefore, this study was conducted to evaluate the effect of hydrolysates from chicken meat and hydrolysates from chicken heart and liver on digestibility, gut health, skin and coat health, oxidative stress, and inflammation in healthy adult dogs. Five canine diets were manufactured with similar ingredients except for the test protein sources. The control diet was formulated with chicken meal, a traditional protein source in pet food, and the other diets had a partial or complete substitution from the chicken meal with the two types of protein hydrolysate. The diets were all well accepted by the dogs and all dogs maintained healthy throughout the study. Dogs fed the hydrolysate from chicken meat showed lower inflammatory biomarkers in the feces. On the other hand, dogs fed the hydrolysate from chicken liver and heart demonstrated a shift in gut microbiota with more abundant beneficial bacteria. In conclusion, poultry-originated protein hydrolysates showed the potential in making positive changes in inflammatory state, immune response, and microbiota in healthy adult dogs.

Functional properties of Ganoderma lucidum supplementation in canine nutrition.

Ganoderma lucidum (GL) is a mushroom that has been widely used in Asia for its immunostimulatory and anti-inflammatory capacity, which has been hypothesized to be attributed mainly to the recognition of its cell-surface patterns by cells of the immune system present in the gastrointestinal tract, resulting in a cascade of modulatory events. However, the nutraceutical properties of GL have not been tested in dogs. Forty adult beagles were used in a completely randomized design. The objective of the present study was to evaluate the effects of dietary inclusion of GL on peripheral blood mononuclear cells (PBMC; T cells, B cells, monocytes, and natural killers), vaccine response, nutrient digestibility, fecal fermentative end-products, and skin and coat quality of adult dogs. Dogs were fed a commercial dry extruded complete and balanced diet plus GL top-dressed daily upon feeding time. Four experimental treatments were used: 0% GL supplementation (control), 5 mg/kg BW of GL, 10 mg/kg BW of GL, or 15 mg/kg BW of GL. Following a 7 d adaptation to the control diet, dogs were fed their respective treatment diets for 28 d. They were challenged with vaccination of a modified live virus Canine Distemper, Adenovirus Type 1 (Hepatitis), Adenovirus Type 2, Parainfluenza, and Parvovirus and killed Rabies Virus on day 7 with blood collections on days 0, 14, and 28. The inclusion of GL in all dosages was well-accepted by all dogs, with no detrimental effect on macronutrient apparent total tract digestibility. There was a trend that the percentage of major histocompatibility II (MHC-II) from B cells was greater in dogs fed 15 mg/kg of GL (41.91%) compared to the control group (34.63%). The phagocytosis response tended to have treatment-by-time interaction among treatments; dogs fed 15 mg/kg of GL tended to have greater phagocytosis activity on day 28 than dogs from the control group and dogs fed 5 mg/kg of GL. The vaccine-specific serum immunoglobulin G (IgG) concentrations were higher in the group supplemented with 15 mg/kg of GL compared to treatment control 7 d after the vaccination for rabies. These data suggest that the inclusion of GL had no detrimental effects on any analyzed PBMC. Due to changes in immune parameters among treatments, GL may also exert beneficial immunostimulatory effects in healthy adult dogs when provided at a daily dose of 15 mg/ kg BW.

Ganoderma lucidum (GL) is a fungus from which products have become popular in the human food and health industry over the past decade. Due to this, a growing interest in using GL extracts in animal products has also developed. The current study investigated the nutritional properties of GL supplemented to adult beagles in three different inclusion levels in terms of body weight (BW; 5, 10, and 15 mg/kg BW). The results indicated no impact on the overall health, apparent total tract macronutrient digestibility (ATTD), fecal microbial DNA, and skin and coat health. The highlighted results included increased phagocytic activity and vaccine-specific response in the group of dogs supplemented with 15 mg/kg BW.

Green banana flour as a novel functional ingredient in retorted feline diets.

Green banana flour (GBF) is a novel ingredient that is high in resistant starch and could be a dietary fiber source in companion animal nutrition. In addition, with its light brown color and pectin content, GBF could potentially serve as a natural color additive and thickening agent in pet food manufacturing. The purpose of this research is to evaluate different sources of GBF, the effect of GBF on texture and color in canned foods, and its effect on apparent total tract digestibility (ATTD), fecal characteristics, and fecal fermentative end-products in healthy adult cats. Prior to the feline study, different sources of GBF were analyzed for chemical composition, manufacturing properties, true metabolizable energy, and fermentability. For the feline feeding trial, all treatment diets were formulated to meet or exceed the Association of American Feed Control Officials (Association of American Feed Control Officials (AAFCO) 2020. Official Publication. Champaign, IL.) guidelines for adult cat maintenance. There were five dietary treatments: rice control (4% rice flour), potato control (4% dehydrated potato flakes), 1% GBF (1% GBF and 3% rice flour), 2% GBF (2% GBF and 2% rice flour), and 4% GBF. All treatment diets were analyzed for texture and color. The animal study was conducted using a completely randomized design with 39 adult domestic cats. There was a 7-d diet adaptation period followed by a baseline fresh fecal collection to determine fecal score, pH, short-chain fatty acid, branched-chain fatty acid, phenol, indole, ammonia, and microbiota. The treatment period lasted for 21 d and a total fecal collection was performed during the last 4 d of this period to determine the ATTD. A fresh fecal sample was also collected during the total fecal collection to evaluate fecal score, pH, metabolites, and microbiota. The MIXED model procedures of SAS version 9.4 were used for statistical analysis. Treatment diets containing GBF had a lower hardness from the texture profile analysis (P < 0.05). For color analysis, the 4% GBF diet was darker in color compared with the rice diet (P < 0.05). There was no difference in food intake, fecal output, or ATTD of macronutrients among the treatment groups (P > 0.05). There was no interaction of treatment and time or main effects shown in fecal score, pH, metabolites, or microbiota diversity (P > 0.05). In conclusion, adding GBF to canned diets may affect the texture and color of the product, but GBF was comparable to traditional carbohydrate sources, rice, and potato, from a nutritional aspect.

Green banana flour (GBF) is a novel ingredient in the pet food industry but has been gaining popularity in human nutrition. Not only can GBF be a source of dietary fiber in pet foods, but the natural brown color and hygroscopic properties also show the potential in contributing to physical characteristics. With its soluble fiber content, green banana flour has fewer calories than a digestible starch and is partially fermentable. The current study aimed to examine the effect of green banana flour on canned cat foods in comparison to traditional starch sources. Canned diets were made with predominately chicken and the test carbohydrate sources of rice flour, dehydrated potato flakes, and/or green banana flour. Canned food with a high inclusion rate of green banana flour showed differences in texture and color when compared with traditional diets; the finding indicated that green banana flour could be utilized to obtain desirable wet food characteristics, including color and texture. As a carbohydrate source in the canned diet, green banana flour had comparable effects on digestibility and gut microbiota to traditional starches when fed to cats. In conclusion, green banana flour can be used as an alternative carbohydrate source in canned diets and contribute to product texture and color.

Standardized amino acid digestibility and protein quality in extruded canine diets containing hydrolyzed protein using a precision-fed rooster assay.

Protein hydrolysate has become a choice of alternative protein source in canine diets as it showed greater digestibility, lower allergenic responses, and various functional properties when compared with intact proteins. The objective of the study was to determine the effect of hydrolyzed protein inclusion on amino acid digestibility and protein quality in extruded canine diets when compared with a traditional protein source for adult dogs. Five treatment diets were formulated to have similar compositions except for the main protein source. The control diet was formulated with chicken meal (CM) as the primary protein source. Test hydrolyzed proteins, chicken liver and heart hydrolysate (CLH) and chicken hydrolysate (CH) were used to partially or completely substitute CM. The diets were: CONd: CM (30%) diet; 5%CLHd: 5% CLH with 25% CM diet; CLHd: CLH (30%) diet; 5%CHd: 5% CH with 25% CM diet; CHd: CH (30%) diet. A precision-fed rooster assay was used to determine standardized amino acid digestibility for the ingredients and diets. In addition, Digestible Indispensable Amino Acid Score (DIAAS)-like values were calculated for the protein ingredients. All protein ingredients had higher than 80% of digestibility for all indispensable amino acids with no difference among sources (P > 0.05). From the DIAAS-like values referencing AAFCO nutrient profile for adult dogs, CLH and CH did not have any limiting amino acid; on the other hand, CM has a lower DIAAS-like value (93.3%) than CLH and CH (P < 0.05) with tryptophan being the first-limiting amino acid. The DIAAS-like values were often lower when the amino acid combinations methionine + cysteine and phenylalanine + tyrosine were included in the calculation. When referencing NRC recommended allowances and minimal requirements, methionine was the first-limiting amino acid for all protein sources. Amino acid digestibility was mostly above 80% and comparable among the treatment diets. Regarding the digestible indispensable amino acid concentrations in the diets, all of them met the AAFCO nutrient profile for adult dogs at maintenance. In conclusion, both protein hydrolysates were highly digestible, high-quality protein sources, and a full substitution from CM to protein hydrolysate could result in greater protein quality, according to the DIAAS-like values of the ingredients, when compared with CM in extruded canine diets.

Hydrolyzed protein has been more commonly used in the pet food industry recently to increase digestibility, decrease allergenic responses, and for other health benefits (such as anti-inflammation and anti-oxidation) in companion animals. The study was designed to determine the protein quality of two protein hydrolysates for incorporating into dry dog foods. Chicken meal (CM) was chosen to be the control protein source as it is a high-quality protein source and has been widely used in pet foods. The test hydrolyzed proteins were made from chicken liver and heart or mechanically separated chicken. The current study aimed to compare the test hydrolyzed proteins, which are usually highly digestible, with a traditional high-quality protein to examine if the hydrolyzed proteins were comparable with CM or of higher quality. Five treatment diets were formulated with CM and/or hydrolyzed proteins. Cecectomized roosters were used in the assay to determine the amino acid digestibility and protein quality of the ingredients and treatment diets. It was shown that all ingredients and diets were highly digestible. The protein hydrolysates were of higher protein quality that better met the animals' needs. Conclusively, the test protein hydrolysates are suitable protein sources in canine diets to create high-value and specialty foods.

The impact of protein source and grain inclusion on digestibility, fecal metabolites, and fecal microbiome in adult canines.

This study was conducted to determine the effect of animal protein inclusion rate and grain-free or grain-inclusive diets on macronutrient digestibility, fecal characteristics, metabolites, and microbiota in mixed-breed hounds and Beagles. Four experimental extruded kibble diets were made with varying amounts of animal protein and carbohydrates: 1) high animal protein, grain-inclusive (HA-GI), 2) low animal protein, grain-free (LA-GF), 3) low animal protein, grain-inclusive (LA-GI), and 4) high animal protein, grain-free (HA-GF). Thirty-two Beagles and 33 mixed-breed hounds were assigned to 1 of the 4 treatment groups in a completely randomized design that lasted 180 d. All diets were similar in chemical composition and well-digested by the animals. In general, for fecal metabolites, mixed-breed hounds had a greater concentration of total short-chain fatty acid (SCFA) and ammonia and lower indole concentration than Beagles (P < 0.05). In mixed-breed hounds, LA-GF had a greater (P < 0.05) total SCFA concentration than HA-GI and LA-GI; however, this was not observed in Beagles. There were greater concentrations of ammonia, phenol, and indole in HA-GI than in LA-GF (P < 0.05). Breed-affected fecal primary bile acid (BA) concentration, as mixed-breed hounds had a greater concentration of cholic acid (CA) than Beagles (P < 0.05). Mixed-breed hounds fed LA-GF resulted in greater CA concentrations than HA-GI and LA-GI (P < 0.05). Dogs who consumed LA-GF had lower fecal secondary BA content than the other groups (P < 0.05). The distribution of the fecal microbiota community differed in LA-GF compared with the other groups, with lower α-diversity. However, dogs fed LA-GF had the largest difference in composition with greater Selenomonadaceae, Veillonellaceae, Lactobacillaceae, Streptococcus, Ligilactobacillus, Megamonas, Collinsella aerofaciens, and Bifidobacterium sp. than the other groups. A significant breed effect was noted on nutrient digestibility, fecal metabolites, and microbiota. A treatment effect was observed in LA-GF as it resulted in greater fecal SCFA, lower protein fermentative end products, greater fecal primary BAs, lower fecal secondary BA concentrations, and shifts in fecal microbiota.

A recent topic of debate in pet food is whether grain or pulse ingredients and varying amounts of animal-based protein compared to plant proteins are better for pets' health. Thus, the need to examine ingredients used in pet food is important. This study aimed to evaluate macronutrient digestibility, fecal characteristics, fecal metabolites, and fecal microbiota in both mixed-breed hounds and Beagles when fed extruded diets containing different inclusion rates of animal protein and plant-based ingredients. Four experimental diets were studied 1) high animal protein, grain-inclusive (HA-GI), 2) low animal protein, grain-free (LA-GF), 3) low animal protein, grain-inclusive (LA-GI), and 4) high animal protein, grain-free (HA-GF). We found that all four diets were well-digested by the animals and all dogs remained healthy throughout the study. In addition, LA-GF produced a decrease in alpha diversity, yet was greater in the abundance of Megamonas, which are known to produce short-chain fatty acids. The other diets did not differ significantly. Apparent total tract nutrient digestibility, fecal characteristics, metabolites, and microbiota were affected by breed and dietary treatments. While some have postulated that pulse-rich diets could perhaps be a cause of nutrition-associated dilated cardiomyopathy in canines due to a potentially negative effect on digestibility, our results showed all diets were highly digestible.

Nutrient profile, amino acid digestibility, true metabolizable energy, and indispensable amino acid scoring of whole hemp seeds for use in canine and feline diets.

The use of various hemp-derived products has been rapidly growing in the human nutrition industry and has sparked great interest in using these ingredients for companion animals as well. Thorough research is needed to determine the ingredient and safety standards required for AAFCO approval of hemp ingredients. In order to be effectively incorporated into pet food products, we must determine the nutrient content, quality, and utility of these ingredients in pet species. The objective of this study was to evaluate the nutrient composition of seeds from four different varieties of hemp, NWG 452, NWG 331, NWG 2730, X-59, and determine protein quality and true metabolizable energy using a cecectomized rooster model. The seeds were similar in macronutrient composition, with small variations in acid hydrolyzed fat, crude protein, total dietary fiber and gross energy content, as well as amino acid and long-chain fatty acid profiles. All essential amino acids were present in concentrations that exceeded the NRC (2006) recommended allowances for adult dogs and cats at maintenance with the exception of tryptophan. The long-chain fatty acid profile presented a favorable ratio of omega-6 to omega-3 fatty acids of close to 3.5:1. The results of the cecectomized rooster assay indicated no significant difference in the standardized amino acid digestibility of the indispensable amino acids among the seed varieties (P > 0.05). A significant difference in the true metabolizable energy corrected for nitrogen was observed among the seeds (P < 0.05), following the pattern of higher acid hydrolyzed fat and lower total dietary fiber content resulting in higher metabolizable energy. An adapted calculation of digestible indispensable amino acid score was made to determine protein quality of the hemp seeds using AAFCO nutrient profiles and NRC recommended allowances for adult dogs and cats at maintenance as reference points. The resulting scores determined tryptophan to be the first limiting amino acid and indicate that hemp seeds alone do not meet all the amino acid requirements for adult dogs and cats at maintenance, and would need a complimentary protein source for practical use in companion animal diets. The data from this study suggest that hemp seeds may provide a beneficial source of fat, protein, and dietary fiber, with consideration to differences in nutrient profile among seed varieties. However, further investigation in vivo is needed to determine the safety and efficacy of utilizing hemp in the diets of both canines and felines.

Hemp products have become popular in the human food and health industry over the past few years. Due to this, a growing interest in using hemp ingredients in animal products has developed as well. There is a need to investigate the nutritional properties and potential utility of hemp seeds in food products for companion animals in order for them to be consumed safely and effectively. Four different varieties of hemp seed were evaluated and found to have similar fat, fiber, and protein content as well as protein quality. The results indicate that hemp seeds may be an advantageous ingredient in the development of pet food products, but a more in depth evaluation using pet species is necessary to confirm this.

Effects of a high-protein, high-fiber diet rich in antioxidants and l-carnitine on body weight, body composition, metabolic status, and physical activity levels of cats after spay surgery.

Spay and neuter surgeries are useful in controlling pet populations, but increase obesity risk due to increased appetite, decreased metabolic rate, and decreased energy expenditure. Dietary management may help limit post-spay weight gain, but few research studies have been conducted in cats. Therefore, the objective of this study was to evaluate the effects of a high-protein, high-fiber diet (HPHF) compared to a moderate-protein, moderate-fiber diet (MPMF) in female cats following spay surgery. Twenty healthy female cats (9.5 ±â€…0.1 mo) were used. After a 4-wk baseline phase with cats fed MPMF to maintain body weight (BW), 16 cats were spayed and allotted to MPMF (n = 8) or HPHF (n = 8), with the remaining cats being sham-operated and fed MPMF (n = 4). Cats were fed to maintain BW for 12 wk and then allowed to eat up to twice that amount for another 12 wk. Daily food intake, twice weekly BW, and twice weekly body condition scores (BCS) were assessed. Back fat thickness (BF) using ultrasound, body composition using dual-energy X-ray absorptiometry (DEXA), feline body mass index (fBMI), body fat percentage estimates using zoometry measurements, serum metabolites, and voluntary physical activity levels were measured prior to spay (week 0) and every 6 wk post-spay. A treatment*time effect was observed for food intake (g/d), but not caloric intake (kcal ME/d). Caloric intake was affected by time and treatment, being reduced over the first 12 wk and reduced at higher amounts in HPHF and MPMF cats vs. sham cats. BW, BCS, and body fat percentage were affected over time. Treatment*time effects were observed for blood urea nitrogen, alkaline phosphatase, and fructosamine, whereas blood triglycerides, total cholesterol, creatinine, total protein, phosphorus, and bicarbonate were affected by time. Physical activity was reduced over time. Our results demonstrate that spay surgery affects food intake, BW, metabolism, and physical activity of cats. Dietary intervention in this study, however, led to minor changes.

Spay surgery helps control pet populations, but increases obesity due to increased appetite, decreased metabolic rate, and decreased energy expenditure. Our objective was to evaluate the effects of high-protein, high-fiber diet (HPHF), and moderate-protein, moderate-fiber diets (MPMF) in female cats following spay surgery. Of the 20 cats used, 16 were spayed and fed MPMF (n = 8) or HPHF (n = 8) and four were sham-operated and fed MPMF. Cats were fed to maintain body weight (BW) for 12 wk and then allowed to overeat for 12 wk. Food intake, BW, body condition scores (BCS), back fat thickness, body composition, feline body mass index, body fat percentage estimates, serum metabolites, and physical activity levels were measured. Over the first 12 wk, caloric intake was reduced at higher amounts in spayed versus sham cats. BW, BCS, body fat percentage, and physical activity levels were altered over time. Our results demonstrate that the diets tested had minor effects, but spaying affected cat food intake, BW, metabolism, and physical activity.

Palatability and apparent total tract macronutrient digestibility of retorted black soldier fly larvae-containing diets and their effects on the fecal characteristics of cats consuming them.

There is a growing interest in using black soldier fly larvae (BSFL) due to its supposed sustainability and nutritional qualities. Because little research has been conducted to evaluate the use of BSFL in cats, our objective was to determine the palatability and apparent total tract macronutrient digestibility (ATTD) of BSFL-containing canned diets and the fecal characteristics of healthy adult cats consuming them. First, three palatability tests were conducted to compare the following diets: 1) diet with poultry byproduct meal (PBPM) and chicken serving as the primary protein sources (control) vs. diet with BSFL meal replacing PBPM (BSFL meal); 2) control vs. diet with whole BSFL replacing some PBPM and poultry fat (BSFL whole); and 3) control vs. diet with BSFL oil replacing poultry fat (BSFL oil). All diets were formulated to meet Association of American Feed Control Officials nutrient profiles for adult cats and were produced using a still retort. A paired t-test was conducted to analyze data from each palatability test, with a higher (P < 0.05) consumption ratio being observed for BSFL meal (1.93:1), BSFL whole (2.03:1), and BSFL oil (1.57:1). Second, 32 adult cats (20 females and 12 males; BW: 4.19 ± 0.55 kg; age: 3.3 ± 0.38 yr) were used in a completely randomized design study composed of a 21-d baseline period and a 70-d experimental period. Cats consumed the control diet during the baseline and were then allotted to one of four experimental diets (n = 8 per group): 1) control, 2) BSFL meal, 3) whole BSFL, and 4) BSFL oil. Fecal samples were collected after baseline and experimental periods for ATTD and fecal characteristic analysis. Fecal output was higher (P < 0.05) and fecal dry matter percentage was lower (P < 0.05) in cats fed BSFL meal than those fed BSFL oil. Organic matter, crude protein (CP), and energy ATTD were lower (P < 0.05) in cats fed BSFL meal than those fed BSFL oil or control. CP and energy ATTD were lower (P < 0.05) in cats fed BSFL whole than those fed BSFL oil. A few serum metabolites were affected by diet (P < 0.05) but remained within reference ranges. Hematology was not affected by diet (P > 0.05). Overall, our results suggest that BSFL-containing diets are palatable and do not negatively affect fecal characteristics or serum chemistry but may have slightly lower nutrient digestibilities in adult cats.

The pet food industry is interested in black soldier fly larvae (BSFL) because of its supposed sustainability and nutritional qualities. BSFL has not been well tested in cats, so our objective was to determine the palatability and apparent total tract macronutrient digestibility (ATTD) of BSFL-containing canned diets and how they affected the fecal characteristics of adult cats. Palatability tests were conducted to compare diets containing BSFL meal, whole BSFL, or BSFL oil against a poultry-based control. Higher consumption ratios were observed for all BSFL-containing diets. A second study determined the digestibility of BSFL-containing diets and how they affected fecal characteristics. Cats were allotted to a control diet or diets containing BSFL meal, whole BSFL, or BSFL oil. Fecal output was higher and fecal dry matter percentage was lower in cats fed BSFL meal than those fed BSFL oil. Organic matter, crude protein (CP), and energy ATTD were lower in cats fed BSFL meal than those fed BSFL oil or control. CP and energy ATTD were lower in cats fed whole BSFL than those fed BSFL oil. Overall, BSFL-containing diets were shown to be palatable and did not negatively affect fecal characteristics, but reduced nutrient digestibilities in adult cats.

Chemical composition of selected insect meals and their effect on apparent total tract digestibility, fecal metabolites, and microbiota of adult cats fed insect-based retorted diets.

Insect meals are novel and potentially sustainable protein sources. The objectives of this study were to determine the chemical composition and standardized amino acid digestibility using the cecectomized rooster model of three selected insect meals (i.e., speckled cockroach [SC], Madagascar hissing cockroach [MC], and superworm [SW]) and to determine the effects of these insect meals on food intake, apparent total tract digestibility (ATTD) of macronutrients, fecal scores, and metabolites of adult cats fed insect- or chicken-based retorted diets. This study consisted of a complete randomized design, with 28 adult cats randomly assigned to one of the four experimental retorted diets: Control (chicken-based diet), SC diet, MC diet, or SW diet. All animal procedures were approved by the University of Illinois Institutional Animal Care and Use Committee. All diets were formulated to be complete and balanced and meet or exceed the nutritional requirements of adult cats. The experimental period was 28 d, with the first 7 d allotted for diet adaptation. The total fecal collection was completed during the last 4 d of the experimental period. On day 21, a fresh fecal sample from each cat was collected for the determination of fecal metabolites and microbiota. Food was offered twice daily to maintain body weight and body condition score. Among the three selected insect meals evaluated, oleic acid, palmitic acid, linoleic acid, and stearic acid were the most prevalent fatty acids. Branched-chain amino acids and arginine were the most preponderant indispensable amino acids in these insect meals. ATTD of dry matter, organic matter, acid-hydrolyzed fat, and crude protein did not differ among treatments (P > 0.05), and all diets were well digested by the cats. Similarly, fecal scores did not differ among the treatments and were within ideal range. No differences (P > 0.05) in fecal metabolite concentrations or microbiota diversity were observed among cats fed different experimental diets; only a few genera from Firmicutes and Bacteroidota phyla differ (P < 0.05) in cats fed SW diet in contrast to other dietary treatments. In conclusion, the selected insect meals evaluated herein are rich in linoleic acid, an essential fatty acid for cats. Insect-based retorted diets led to comparable results to those achieved with a chicken-based retorted diet, suggesting that these novel protein sources might be adequate alternative ingredients in feline diets.

Insect meals are novel and potentially sustainable protein sources. The objectives of this study were to determine the nutrient composition of speckled cockroach, Madagascar hissing cockroach, and superworm (SW) and to determine the effects of these insect meals on food intake, digestibility of macronutrients, fecal scores, metabolites, and microbiota of adult cats fed insect- or chicken-based wet pet foods. Among the three selected insect meals evaluated, oleic acid, palmitic acid, linoleic acid, and stearic acid were the most prevalent fatty acids. Branched-chain amino acids and arginine were the most preponderant indispensable amino acids in these insect meals. All diets were well digested by the cats with no differences observed on macronutrient digestibility. Similarly, fecal scores did not differ among the treatments and were within the ideal range. No differences in fecal metabolite concentrations were observed. Only a few genera from Firmicutes and Bacteroidota phyla differ in cats fed SW diet in contrast to other dietary treatments. Overall, the selected insect meals evaluated herein are rich in linoleic acid, an essential fatty acid for cats. Insect-based retorted diets led to comparable results to those achieved with a chicken-based retorted diet, suggesting that these novel protein sources might be adequate alternative ingredients in feline diets.

Longitudinal assessment of taurine and amino acid concentrations in dogs fed a green lentil diet.

A recent association between the inclusion of pulses in canine diets and taurine deficiency has become a prevalent issue in the pet food industry. Although dogs do not currently have a nutritional requirement for taurine, taurine deficiencies that do occur can result in serious health issues, such as dilated cardiomyopathy. The objective of this study was to determine the circulating concentrations of plasma and whole blood taurine, indispensable and dispensable amino acid concentrations in the plasma, and taurine and creatinine concentrations in the urine of adult dogs fed a green lentil diet. Twelve adult, intact, female beagles were randomly assigned to a diet containing 45% green lentils (GLD) or a poultry byproduct meal diet (CON) for 90 d. Fresh urine samples were collected every 30 d and analyzed for taurine and creatinine concentrations. A blood sample was also collected every 30 d and analyzed for amino acids including taurine. Animal procedures were approved by the University of Illinois Institutional Animal Care and Use Committee. All diets were formulated to meet or exceed the nutrient requirements for adult dogs at maintenance. The concentrations of taurine in the plasma and whole blood showed no differences (P > 0.05) between dietary treatments or across time points. Similarly, no differences (P > 0.05) in plasma methionine concentrations were observed between treatments or across time points. A treatment effect (P < 0.05) showed dogs fed GLD had higher total primary fecal bile acid excretion compared with dogs fed CON. The differential abundance of fecal microbial communities showed Firmicutes as the predominant phyla in dogs fed both GLD and CON, with Bacteroidaceae, Erysipelotrichaceae, and Lactobacillaceae as predominant families in dogs fed GLD. The α-diversity of dogs fed GLD (P < 0.05) was lower than in dogs fed CON. These data suggest that the inclusion of 45% green lentil in extruded diets does not lower whole blood and plasma taurine concentrations during a 90-d period and is appropriate for use in a complete and balanced formulation for dogs.

Effects of graded inclusion levels of raw garbanzo beans on apparent total tract digestibility, fecal quality, and fecal fermentative end-products and microbiota in extruded feline diets.

Garbanzo beans (GB; Cicer arietinum) are a readily available pulse crop that have gained popularity as a plant-based protein source in the pet food industry. However, raw GB contain anti-nutritional factors that can reduce digestibility and cause digestive upsets in pets that are undesirable to owners. The objective of this study was to determine the effects of the inclusion of raw or cooked GB in extruded feline diets on macronutrient digestibility, gastrointestinal tolerance, and fermentative end-products in cats. Five diets were formulated to contain raw GB at 0%, 7.5%, 15%, or 30% or cooked GB at 30%. Ten adult, male cats (mean age: 1.0 ± 0.0 yr, mean BW: 4.7 ± 0.4 kg) were used in a replicated 5 × 5 Latin square design. Each period consisted of 14 d, with 10 d of diet adaptation followed by 4 d of total fecal and urine collection. At the end of each period, 4 mL of blood were collected and analyzed for a serum chemistry and complete blood count to ensure all animals remained healthy throughout the study. Cats were fed twice daily and food intake was calculated to maintain body weight. Food intake was highest (P < 0.05) for cats fed 0% raw GB (72.2 g/d, dry matter basis [DMB]) compared with GB inclusions of 7.5% or greater (average 70.3 g/d, DMB). Dry matter and organic matter apparent total tract digestibility (ATTD) were lowest (P < 0.05) for cats consuming the 30% cooked GB diet (77.3% and 81.7%, respectively). Cats fed 7.5% raw GB had greater (P < 0.05) crude protein ATTD (86.2%) than cats fed 15% raw GB (82.3%) or 30% cooked GB (81.6%). Total short-chain fatty acid concentrations were highest (P < 0.05) for 30% cooked GB at 682 µmol/g but not different (P > 0.05) than 15% GB (528 µmol/g) or 30% raw GB (591 µmol/g) diets. In terms of fecal microbial abundance, the predominant phyla were Firmicutes, Bacteroidota, and Actinobacteria. Cats fed the 0% GB diet had a greater relative abundance of Firmicutes (62.1%) and Fusobacteria (4.0%) than the remaining diets (average 54% and 1.6%, respectively). In conclusion, all inclusion levels of raw GB resulted in high digestibility (average > 80%) and ideal fecal scores (average 2.9), demonstrating their adequacy as a protein source in feline diets up to a 30% inclusion level.

Effect of crude fiber and total dietary fiber on the calculated nitrogen-free extract and metabolizable energy content of various dog foods fed to client-owned dogs with osteoarthritis.

OBJECTIVE: To compare measurements of crude fiber (CF) and total dietary fiber (TDF) for various dog foods and their effect on the calculated nitrogen-free extract and metabolizable energy (ME) content, and to compare label-guaranteed and laboratory-analyzed macronutrient values. SAMPLES: 51 dog foods fed to client-owned dogs with osteoarthritis. PROCEDURES: Foods were analyzed for dry matter, ash, crude protein, acid-hydrolyzed fat, CF, and TDF. Metabolizable energy was calculated by use of a formula with modified Atwater factors and formulas recommended by the National Research Council that included both CF and TDF values. Linear regression analysis was performed to determine the correlation between CF and TDF values. RESULTS: Only a few foods failed to conform to the guaranteed analysis for all macronutrients except for CF, in which approximately 40% of the foods exceeded the guaranteed maximum values. The CF and TDF values were moderately correlated (r = 0.843). Correlations among CF- and TDF-based ME estimations were moderate with use of the modified Atwater formula and strong with use of the National Research Council formulas (r = 0.86 and r = 0.91, respectively). CONCLUSIONS AND CLINICAL RELEVANCE: Values for CF were the most variable of the macronutrients of the evaluated dog foods and results suggested that CF is an incomplete and inaccurate measurement of dietary fiber content and, thus, its inaccuracy may lead to inaccurate and variable ME values.

Subacute Exposure to an Environmentally Relevant Dose of Di-(2-ethylhexyl) Phthalate during Gestation Alters the Cecal Microbiome, but Not Pregnancy Outcomes in Mice.

Di-2-ethylhexyl phthalate (DEHP) is a plasticizer commonly found in polyvinyl chloride, medical equipment, and food packaging. DEHP has been shown to target the reproductive system and alter the gut microbiome in humans and experimental animals. However, very little is known about the impact of DEHP-induced microbiome changes and its effects during pregnancy. Thus, the objective of this study was to investigate the effects of DEHP exposure during pregnancy on the cecal microbiome and pregnancy outcomes. Specifically, this study tested the hypothesis that subacute exposure to DEHP during pregnancy alters the cecal microbiome in pregnant mice, leading to changes in birth outcomes. To test this hypothesis, pregnant dams were orally exposed to corn oil vehicle or 20 µg/kg/day DEHP for 10 days and euthanized 21 days after their last dose. Cecal contents were collected for 16S Illumina and shotgun metagenomic sequencing. Fertility studies were also conducted to examine whether DEHP exposure impacted birth outcomes. Subacute exposure to environmentally relevant doses of DEHP in pregnant dams significantly increased alpha diversity and significantly altered beta diversity. Furthermore, DEHP exposure during pregnancy significantly increased the relative abundance of Bacteroidetes and decreased the relative abundance of Firmicutes and Deferribacteres compared with controls. The affected taxonomic families included Deferribacteraceae, Lachnospiraceae, and Mucisprillum. In addition to changes in the gut microbiota, DEHP exposure significantly altered 14 functional pathways compared with the control. Finally, DEHP exposure did not significantly impact the fertility and birth outcomes compared with the control. Collectively, these data indicate that DEHP exposure during pregnancy shifts the cecal microbiome, but the shifts do not impact fertility and birth outcomes.

Effects of dietary macronutrient profile on apparent total tract macronutrient digestibility and fecal microbiota, fermentative metabolites, and bile acids of female dogs after spay surgery.

Obesity and estrogen reduction are known to affect the gut microbiota and gut microbial-derived metabolites in some species, but limited information is available in dogs. The aim of this study was to determine the effects of dietary macronutrient profile on apparent total tract macronutrient digestibility, fecal microbiota, and fecal metabolites of adult female dogs after spay surgery. Twenty-eight adult intact female beagles (age: 3.02 ± 0.71 yr, BW: 10.28 ± 0.77 kg; BCS: 4.98 ± 0.57) were used. After a 5-wk baseline phase (week 0), 24 dogs were spayed and randomly allotted to one of three experimental diets (n = 8 per group): 1) control (CO) containing moderate protein and fiber (COSP), 2) high-protein, high-fiber (HPHF), or 3) high-protein, high-fiber plus omega-3 and medium-chain fatty acids (HPHFO). Four dogs were sham-operated and fed CO (COSH). All dogs were fed to maintain BW for 12 wk after spay and then allowed to consume twice that amount for 12 wk. Fecal samples were collected at weeks 0, 12, and 24 for digestibility, microbiota, and metabolite analysis. All data were analyzed using repeated measures and linear mixed models procedure of SAS 9.4, with results reported as a change from baseline. Apparent organic matter and energy digestibilities had greater decreases in HPHF and HPHFO than COSH and COSP. Increases in fecal acetate, total short-chain fatty acids, and secondary bile acids were greater and decreases in primary bile acids were greater in HPHF and HPHFO. Principal coordinates analysis of weighted UniFrac distances revealed that HPHF and HPHFO clustered together and separated from COSH and COSP at weeks 12 and 24, with relative abundances of Faecalibacterium, Romboutsia, and Fusobacterium increasing to a greater extent and Catenibacterium, Bifidobacterium, Prevotella 9, Eubacterium, and Megamonas decreasing to a greater extent in HPHF or HPHFO. Our results suggest that high-protein, high-fiber diets alter nutrient and energy digestibilities, fecal metabolite concentrations, and fecal gut microbiota, but spay surgery had minor effects. Future research is needed to investigate how food intake, nutrient profile, and changes in hormone production influence gut microbiota and metabolites of dogs individually and how this knowledge may be used to manage spayed pets.

Use of the precision-fed cecectomized rooster assay to determine standardized amino acid digestibility, true metabolizable energy content, and digestible indispensable amino acid scores of plant-based protein by-products used in canine and feline diets.

Traditionally, protein by-products from oil seeds and cereal grains have been used in pet foods as sustainable, inexpensive, and protein-rich ingredients. However, the on-going demonization of soy- and corn-based ingredients continue to hinder their use in pet food and treat formulations. Ideally, the further demonstration of their protein quality and nutrient composition may encourage their favorable return as acceptable ingredients in pet foods and treats. Therefore, the objectives of this study were to determine the macronutrient composition, indispensable amino acid profile, standardized amino acid digestibility, true metabolizable energy content corrected for nitrogen (TMEn), and digestible indispensable amino acid scores (DIAAS-like) of soy flakes (SF), peanut flour (PF), soybean meal (SBM), and corn gluten meal (CGM). Standardized amino acid digestibility was assessed using the precision-fed cecectomized rooster assay. All test ingredients demonstrated a profile of highly digestible indispensable amino acids except for lysine in PF, which was lowest (P < 0.05) at 45.5%. The SBM and CGM had the highest (P > 0.05) digestibilities of indispensable amino acids. A DIAAS-like value was calculated for each ingredient using either AAFCO (2020) recommended values or NRC (2006) recommended allowances as the reference protein pattern. For adult dogs compared to AAFCO recommended values, the first-limiting amino acid was lysine for PF and CGM but it was methionine for SF and SBM. For adult cats compared to AAFCO recommended values, the first-limiting amino acid was lysine for PF and CGM but it was threonine for SF. There was no first-limiting amino acid in SBM for cats as DIAAS-like values were over 100% for all indispensable amino acids. The TMEn values were highest (P < 0.05) for PF and CGM (4.58 and 4.31 kcal/g [dry matter basis], respectively). The protein quality of these plant-based protein by-products reflects their value as nutritional ingredients for canine and feline diets. However, the prior processing of these by-products must be considered before exposing them to additional processing methods, such as extrusion. Additionally, the inclusion of complementary proteins or supplemental amino acids will be needed to meet all indispensable amino acid requirements for a portion of nutritionally complete and balanced pet food.

Physical activity patterns of free living dogs diagnosed with osteoarthritis.

Osteoarthritis (OA) affects about 90% of dogs > 5 yr of age in the United States, resulting in reduced range of motion, difficulty climbing and jumping, reduced physical activity, and lower quality of life. Our objective was to use activity monitors to measure physical activity and identify how activity counts correlate with age, body weight (BW), body condition score (BCS), serum inflammatory markers, veterinarian pain assessment, and owner perception of pain in free-living dogs with OA. The University of Illinois Institutional Animal Care and Use Committee approved the study and owner consent was received prior to experimentation. Fifty-six client-owned dogs (mean age = 7.8 yr; mean BCS = 6.1) with clinical signs and veterinary diagnosis of OA wore HeyRex activity collars continuously over a 49-d period. Blood samples were collected on day 0 and 49, and dog owners completed canine brief pain inventory (CBPI) and Liverpool osteoarthritis in dogs (LOAD) surveys on day 0, 21, 35, and 49. All data were analyzed using SAS 9.3 using repeated measures and R Studio 1.0.136 was used to generate Pearson correlation coefficients between data outcomes. Average activity throughout the study demonstrated greater activity levels on weekends. It also showed that 24-h activity spiked twice daily, once in the morning and another in the afternoon. Serum C-reactive protein concentration was lower (P < 0.01) at day 49 compared to day 0. Survey data indicated lower (P < 0.05) overall pain intensity and severity score on day 21, 35 and 49 compared to day 0. BW was correlated with average activity counts (P = 0.02; r = -0.12) and run activity (P = 0.10; r = -0.24). Weekend average activity counts were correlated with owner pain intensity scores (P = 0.0813; r = -0.2311), but weekday average activity count was not. Age was not correlated with total activity count, sleep activity, or run activity, but it was correlated with scratch (P = 0.03; r = -0.10), alert (P = 0.03; r = -0.13), and walk (P = 0.09; r = -0.23) activities. Total activity counts and activity type (sleep, scratch, alert, walk, and run) were not correlated with pain scored by veterinarians, pain intensity or severity scored by owners, or baseline BCS. Even though the lack of controls and/or information on the individual living conditions of dogs resulted in a high level of variability in this study, our data suggest that the use of activity monitors have the potential to aid in the management of OA and other conditions affecting activity (e.g., allergy; anxiety).

Effects of Weight Loss and Moderate-Protein, High-Fiber Diet Consumption on the Fasted Serum Metabolome of Cats.

Feline obesity elicits a plethora of metabolic responses leading to comorbidities, with potential reversal during weight loss. The specific metabolic alterations and biomarkers of organ dysfunction are not entirely understood. Untargeted, high-throughput metabolomic technologies may allow the identification of biological components that change with weight status in cats, increasing our understanding of feline metabolism. The objective of this study was to utilize untargeted metabolomic techniques to identify biomarkers and gain mechanistic insight into the serum metabolite changes associated with reduced food intake and weight loss in overweight cats. During a four-wk baseline period, cats were fed to maintain body weight. For 18 wk following baseline, cats were fed to lose weight at a rate of ~1.5% body weight/wk. Blood serum metabolites were measured at wk 0, 1, 2, 4, 8, 12, and 16. A total of 535 named metabolites were identified, with up to 269 of them being altered (p- and q-values < 0.05) at any time point. A principal component analysis showed a continual shift in metabolite profile as weight loss progressed, with early changes being distinct from those over the long term. The majority of lipid metabolites decreased with weight loss; however, ketone bodies and small lipid particles increased with weight loss. The majority of carbohydrate metabolites decreased with weight loss. Protein metabolites had a variable result, with some increasing, but others decreasing with weight loss. Metabolic mediators of inflammation, oxidative stress, xenobiotics, and insulin resistance decreased with weight loss. In conclusion, global metabolomics identified biomarkers of reduced food intake and weight loss in cats, including decreased markers of inflammation and/or altered macronutrient metabolism.

Use of Legumes and Yeast as Novel Dietary Protein Sources in Extruded Canine Diets.

The popularity of plant-based protein sources has increased as consumer demand for grain-free and novel protein sources increase. Minimal research has been conducted as regards to use of legumes and yeast and their effects on acceptability and digestibility in canine diets. The objective of this study was to evaluate macronutrient apparent total tract digestibility (ATTD), gastrointestinal tolerance, and fermentative end-products in extruded, canine diets. Five diets were formulated to be isocaloric and isonitrogenous with either garbanzo beans (GBD), green lentils (GLD), peanut flour (PFD), dried yeast (DYD), or poultry by-product meal (CON) as the primary protein sources. Ten adult, intact, female beagles (mean age: 4.2 ± 1.1 yr, mean weight: 11.9 ± 1.3 kg) were used in a replicated, 5 × 5 Latin square design with 14 d periods. Each experimental period consisted of 10 d of diet adaptation, followed by 4 d of total fecal and urine collection. A fasted, 5 ml blood sample was collected at the end of each period and analyzed for serum metabolites and complete blood count. Serum metabolites were within normal ranges and all dogs remained healthy throughout the study. Fecal quality, evaluated on a 5-point scale, was considered ideal. Macronutrient ATTD was similar among dietary treatments, with diets highly digestible (>80%). Total fecal branched-chain fatty acid concentrations were highest (P < 0.05) for DYD (23.4 µmol/g) than GLD (16.1 µmol/g) and PFD (16.0 µmol/g) but not different (P > 0.05) than other treatments. The plant-based protein treatments had greater (P < 0.05) total fecal short chain fatty acid (SCFA) concentrations (average 627.6 µmol/g) compared with CON (381.1 µmol/g). Fecal butyrate concentration was highest (P < 0.05) for DYD than all other dietary treatments (103.9 µmol/g vs. average 46.2 µmol/g). Fecal microbial communities showed Firmicutes, Bacteroidetes, Fusobacteria, and Proteobacteria as abundant phyla. There was greater ß-diversity for dogs fed DYD which differed from all other diets in both weighted and unweighted UNIFRAC analyses. Inclusion of these novel, plant-based, protein sources showed no detrimental effects on nutrient digestibility or fecal characteristics and represent viable protein sources in canine diets that can produce beneficial shifts in fecal metabolites.

Miscanthus Grass as a Novel Functional Fiber Source in Extruded Feline Diets.

Although dietary fiber is not considered an essential nutrient in a complete and balanced diet for felines, it provides a substrate for fermentation by gut microbiota, thus promoting gastrointestinal health through the production of fermentative metabolites, as well as improving laxation. The aim of this research was to evaluate the novel fiber source, Miscanthus grass (Miscanthus giganteus), in comparison with traditional fiber sources and their effects on fecal quality, apparent total tract digestibility (ATTD), fecal fermentative end products, and microbiota of healthy adult cats. Four dietary treatments were evaluated, differing in dietary fiber source. The diets were formulated to meet or exceed the AAFCO (2018) nutritional profile for adult cats and contained either cellulose (CO), Miscanthus grass fiber (MF), a blend of Miscanthus fiber and tomato pomace (MF + TP), or beet pulp (BP). The study was conducted using a completely randomized design with 28 neutered adult, domesticated shorthair cats (19 females and 9 males, mean age 2.2 ± 0.03 years; mean body weight 4.6 ± 0.7 kg, mean body condition score 5.6 ± 0.6). The experimental period comprised 21 days, and a fresh fecal and a total fecal collection were performed during the last 4 days of the trial period. Daily food intake (DM basis) was similar across all groups (P > 0.05). Additionally, treatment did not affect fecal output (as-is or DM basis), fecal score, or fecal pH (P > 0.05). Cats fed BP had significantly higher total dietary fiber ATTD than all the other treatments (P < 0.05) and the highest concentrations of total short-chain fatty acid, acetate, and propionate (P < 0.05), while butyrate concentrations were similar for all treatments (P > 0.05). Inclusion of dietary fibers was effective in modulating gut microbiota. Cats fed diets containing Miscanthus grass had greater α-diversity than cats fed BP. As no adverse effects on health, fecal quality, or ATTD of macronutrients were observed with the inclusion of 9% Miscanthus grass fiber or fiber blend, the data suggest that Miscanthus grass fiber and fiber blends are viable alternatives to the traditional dietary fiber sources used in commercial extruded feline diets, being most comparable to cellulose.

White and Red Sorghum as Primary Carbohydrate Sources in Extruded Diets of Felines.

The research objectives were to evaluate the effect of dietary supplementation of white (WSH) and red (RSH) sorghum grains on gastrointestinal health of felines through the determination of apparent total tract macronutrient digestibility (ATTD), fecal characteristics, fermentative end-products, and microbiota, compared with a traditional corn-based diet. We hypothesize that inclusion of RSH and WSH, respectively, would be well-accepted by cats, and the RSH and WSH diets would be comparable to corn when added as the main carbohydrate source in extruded diets. Three diets containing 30% corn, 30% WSH, or 30% RSH were formulated to meet or exceed the AAFCO (2018) nutrient profiles for cats during growth. Nine male cats (0.8 ± 0.00 yr) were randomly assigned to one of the three dietary treatments using a triplicated 3 × 3 Latin square design. Experimental periods consisted of 14 d (10 d of diet adaption and 4 d of total and fresh fecal collections). The ATTD of dry matter (DM) did not differ amongst treatments, organic matter was greatest (P < 0.05) for both sorghum diets (86.4%) and lowest for the corn diet (84.2%), crude protein was comparable among diets ranging from 84.5 to 86.6%, acid hydrolyzed fat was high among diets varying between 91.4 and 92.8%, and total dietary fiber was greatest (P < 0.05) for the WSH diet (56.0%) with the corn diet being lowest (44.7%). Digestible energy was greatest (P < 0.05) for the WSH diet (4.66 kcal/g) and lowest for the corn diet (4.54 kcal/g), with the RSH diet being intermediate (4.64; P > 0.05). Fecal pH (6.3-6.5) and most fecal metabolites did not differ among diets except for phenol/indole concentrations that were significantly lower (P < 0.05) in cats fed the RSH diet (1.5 µmole/g DM) than for cats fed the corn diet (2.1 µmole/g DM). Bacteroidetes, Firmicutes, Fusobacteria, and Proteobacteria were the major phyla observed in the microbiota of feces of cats fed the three experimental diets, with no differences seen amongst all treatments. Data indicate that dietary supplementation of these varieties of WSH and RSH as carbohydrate sources were well-tolerated by the cat.