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.

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.

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.

Ancient grains as novel dietary carbohydrate sources in canine diets.

Ancient grains are becoming an increasingly abundant carbohydrate source in the pet food market as a result of their popularity and novelty in the human market. Thus, it is imperative to evaluate the characteristics of these ingredients in vivo. Ten adult intact female beagles were used in a replicated 5 × 5 Latin square design. Five dietary treatments were evaluated containing either: rice (CON), amaranth (AM), white proso millet (WPM), quinoa (QU), or oat groats (OG). All diets were formulated to include 40% of the test grain and to be isonitrogenous, isocaloric, and nutritionally complete and balanced for adult dogs at maintenance. The objectives were 1) to evaluate the effects of the novel carbohydrate sources on total apparent total tract digestibility (ATTD), fecal microbiota, and fermentative end-product concentrations and 2) to evaluate the effects of novel carbohydrate sources on the postprandial glycemic and insulinemic responses in healthy adult dogs. All diets were well accepted by the dogs and fecal scores remained within the ideal range for all treatments. In terms of ATTD, all diets were well digested by the dogs; WPM had the highest digestibility of dry and organic matter in contrast with dogs fed the other treatments (P < 0.05). Additionally, ATTD of total dietary fiber was highest for WPM (72.6%) in contrast with QU (63.5%) and CON (50.8%) but did not differ from AM (65.7%) and OG (66.6%). Dogs fed AM or OG had greater (P < 0.05) fecal concentrations of total short-chain fatty acids, as well as propionate and butyrate concentrations, than CON. Ancient grain inclusion appears to beneficially shift fecal microbial populations, with increases in relative abundances of butyrogenic bacteria (i.e., members of the Lachnospiraceae family) observed for OG and reductions in Fusobacteriaceae for both AM and OG when compared with CON. Postprandial glycemic and insulinemic responses did not differ among treatments. Together, these data suggest that ancient grains can be included up to 40% of the diet while eliciting beneficial effects on the overall host health without detrimentally affecting nutrient digestibility.

Chemical composition and in vitro fermentation characteristics of legumes using canine fecal inoculum.

Legumes are a popular grain-free alternative carbohydrate source in canine diets, however, information on their fermentative characteristics have not been established. Thus, the objectives of the present study were to 1) quantify the chemical compositions and 2) fermentative profile of select legumes using canine fecal inoculum. Five legume varieties, whole yellow peas (WYP), green lentils (GL), black bean grits (BBG), navy bean powder (NBP), and garbanzo beans, were analyzed and compared to a positive control, beet pulp (BP). Substrates were analyzed for gross energy (GE), dry and organic matter, crude protein (CP), acid hydrolyzed fat, and total dietary fiber (TDF) fractions, beta-glucans, starch-free, and hydrolyzed sugars, as well as fermentative characteristics: pH, short-chain fatty acids (SCFA), branched-chain fatty acids (BCFA), total gas, hydrogen, and methane. Substrates then underwent a two-stage in vitro digestion and subsequent fermentation using canine fecal inoculum for 0, 3, 6, 9, and 12 h. All test substrates contained approximately 8% to 9% moisture and 4.5 kcal/g GE. The highest CP content was observed in GL (27%). Analyzed TDF content of test substrates was greatest for WYP (32%) and GL (36%). Total starch content was greatest for GL (58%) and WYP (56%). Sucrose and stachyose were the most predominant free sugars and glucose was the most predominant hydrolyzed sugar among test substrates. After 3 and 6 h of fermentation, a net negative change in pH was observed among most substrates with a net negative change in all substrates after 9 and 12 h. Values for SCFA did not differ among substrates after 3 or 6 h of fermentation with BP and WYP among the greatest acetate (1,656 and 1,765 umol/g, respectively) and propionate production values (157.7 and 126.1, respectively) after 9 h. All substrates produced greater total gas volumes than WYP after 3 h, with no differences observed after any other time points. However, BP hydrogen production values were greater after 9 and 12 h (P < 0.0001; 726,042 and 394,675 ng/g, respectively) with greater methane production values after 12 h (P < 0.0001; 54,291 ng/g) than all test substrates. These data suggest that legumes offer a diverse macronutrient profile and appear to be a source of slowly fermentable fiber, which may have beneficial implications on the ratios of saccharolytic to proteolytic fermentation toward the distal colon.

Chemical composition and in vitro fermentation characteristics of ancient grains using canine fecal inoculum.

Human interest in ancient grains replacing traditional carbohydrate sources has reached the pet food market; however, chemical composition of these grains and their digestive properties in the canine model, specifically the fermentative characteristics, have not been established. Five ancient grain varieties were analyzed: amaranth (AM), white proso millet (WPM), oat groats (OG), quinoa (QU), and red millet (RM). Cellulose (CEL) was used as a negative control, and beet pulp (BP) was used as a positive control. Substrates were analyzed for macronutrient composition as well as free and hydrolyzed sugar profiles in addition to their in vitro fermentative characteristics. Substrates were allocated into 2 sets to allow for quantification of pH, short-chain fatty acids, and branched-chain fatty acids, as well as gas volume and composition. Samples were digested for 6 and 18 h with pepsin and pancreatin, respectively, prior to inoculation with fecal bacteria for 0, 3, 6, 9, or 12 h. Detectable levels of cereal ß-glucans were observed solely in OG (3.5%), with all other substrate containing <0.35% cereal ß-glucans. All test substrates had fairly similar macronutrient and starch profiles with the exception of RM that contained the highest resistant starch content (2.4%), with all other test substrates containing <0.5% resistant starch. However, the analyzed pseudocereals, AM and QU, had the highest concentrations of free glucose while the minor cereal grains, WPM, OG, and RM, contained the highest concentrations of hydrolyzed glucose. All test substrates had propionate production values similar or greater than BP after 3, 6, 9, and 12 h of fermentation, and similar or greater butyrate production values than BP after 6, 9, and 12 h. All substrates had greater (P < 0.05) changes in pH than CEL after 6, 9, and 12 h, with AM, WPM, OG, and RM having greater (P < 0.05) changes in pH than BP after 9 and 12 h. These data suggest select ancient grains have similar fermentation characteristics as BP, a moderately fermentable fiber considered the gold standard in terms of fiber sources in the pet food market today, and that OG and AM may be more fermentable during longer fermentation periods.

Use of precision-fed cecectomized rooster assay and digestible indispensable amino acid scores to characterize plant- and yeast-concentrated proteins for inclusion in canine and feline diets.

Increased consumer interest in high-quality and novel protein sources has driven the demand for the inclusion of protein-rich ingredients in companion animal diets. Novel protein concentrates, with protein contents of at least 50%, have been used to satisfy these consumer demands. However, minimal information is available regarding the macronutrient composition and protein quality of these ingredients that is needed for proper formulation of pet foods. Therefore, the objectives of this study were to determine the macronutrient and amino acid compositions, standardized amino acid digestibility according to the precision-fed rooster assay, and protein quality using digestible indispensable amino acid score (DIAAS like) of pea protein (PP), potato protein (POP), faba bean protein (FBP), soy protein concentrate (SPC), and dried yeast (DY). Precision-fed rooster assays were conducted using cecectomized roosters to calculate standardized amino acid digestibility and true metabolizable energy corrected for nitrogen (TMEn). For all five protein concentrates, all essential amino acids were highly digestible (88.0% to 96.3%, dry matter basis) with differences (P < 0.05) in only lysine, methionine, and tryptophan digestibilities. The TMEn values were highest for POP (4.22 kcal/g) and DY (3.61 kcal/g). The DIAAS-like values for adult dogs indicated that methionine was the first-limiting amino acid in all protein concentrates except POP, where the first-limiting amino acid was tryptophan. Using Association of American Feed Control Officials (AAFCO)-recommended values for adult cats, DIAAS-like values for methionine were lowest (P < 0.05) for FBP at 81.5%, with all other amino acids for all protein concentrates over 100%. The National Research Council (NRC)-recommended allowances for adult cats indicated that DIAAS-like methionine values for PP (92.7%) and FBP (73.8%) were significantly lower (P < 0.05) with these being the first-limiting amino acids, with the remaining amino acids above 100% for the other protein concentrates. The protein quality and high essential amino acid digestibility of these protein concentrates indicate that they would be viable protein sources in canine and feline diets. However, additional complementary protein sources should be included to meet the requirements of all essential amino acids.

Macronutrient composition, true metabolizable energy and amino acid digestibility, and indispensable amino acid scoring of pulse ingredients for use in canine and feline diets.

The rising consumer demand for alternative and sustainable protein sources drives the popularity of the use of plant-based proteins in the pet food industry. Pulse crops, which include beans, peas, lentils, and chickpeas, have become an important addition to both human and animal diets due to their protein content and functional properties. However, knowledge of their nutrient composition and protein quality is necessary for the proper formulation of these ingredients in pet foods. The objective of this study was to determine the macronutrient composition and standardized amino acid digestibility and to describe the protein quality through the use of digestible indispensable amino acid scores (DIAAS-like) of five pulse ingredients. Black bean (BB) grits, garbanzo beans (GB), green lentils (GL), navy bean (NB) powder, and yellow peas (YP) were analyzed for dry matter (DM), ash and organic matter (OM), crude protein (CP), gross energy (GE), acid hydrolyzed fat (AHF), and total dietary fiber (TDF) to determine the macronutrient composition. Precision-fed rooster assays were conducted using cecectomized roosters to calculate standardized amino acid digestibility and true metabolizable energy corrected for nitrogen (TMEn). The essential amino acids, with the exception of methionine, were highly digestible with digestibility values of 80% to 90% (dry matter basis) for all selected pulse ingredients. BB grits had the lowest (P < 0.05) digestibility of arginine (86.5%) and histidine (80.6%) in contrast to GB (94.9% and 89.9%, respectively). The TMEn of GB was highest (P < 0.05) at 3.56 kcal/g compared with the other pulses. The DIAAS-like values for adult dogs were consistently the lowest for methionine for all pulses, making it the first-limiting amino acid in these ingredients. The DIAAS-like values for adult cats showed GL had lowest (P < 0.05) score in tryptophan compared with other pulses when using both AAFCO values and NRC recommended allowances as reference proteins. Methionine was the first-limiting amino acid for YP and tryptophan for GL. Based on macronutrient composition, protein quality, and amino acid digestibility, it can be concluded that pulse ingredients have the required nutritional characteristics to be viable protein sources in canine and feline foods. However, the use of complementary protein sources is recommended to counterbalance any potential limiting amino acids in pulse ingredients.