Standardized amino acid digestibility and nitrogen-corrected true metabolizable energy of frozen and freeze-dried raw dog foods using precision-fed cecectomized and conventional rooster assays.

Commercial raw or minimally-processed diets, often referred to holistically as raw meat-based diets (RMBD) represent a small portion of the pet food market, but the growth of this sector has been significant in recent years. While traditionally, high-moisture, frozen options were the standard format of commercially available raw diets, freeze-dried raw diets have become more prevalent as of late. Despite the increasing popularity of these commercial raw diet formats, there is a dearth of literature describing their nutritional properties, particularly regarding freeze-dried diets. Therefore, the objective of this experiment was to determine and compare the standardized amino acid (AA) digestibilities and nitrogen-corrected true metabolizable energy (TMEn) of raw frozen and freeze-dried dog foods using precision-fed cecectomized and conventional rooster assays. Three formats of frozen or freeze-dried raw diets provided by Primal Pet Foods (Fairfield, CA, USA) were tested: traditional freeze-dried nuggets (T-FDN), hybrid freeze-dried nuggets (H-FDN), and frozen nuggets (FZN). Diets were fed to cecectomized roosters (4 roosters/diet) to determine AA digestibilities, while conventional roosters (4 roosters/diet) were used to determine TMEn. In both cases, after 26 h of feed withdrawal, roosters were tube-fed 12 to 13 g of test diets and 12 to 13 g of corn. Following crop intubation, excreta were collected for 48 h. Endogenous corrections for AA were made using five additional cecectomized roosters. All data were analyzed using the Mixed Models procedure of SAS version 9.4. There were no significant differences in standardized AA digestibilities among diets, with digestibilities being high for all diets tested. For most of the indispensable AA, digestibilities were greater than or equal to 90% for all diets. Histidine and lysine were the exceptions, with digestibilities ranging from 82% to 87% and 87% to 92%, respectively. Moreover, the reactive lysine:total lysine ratio, a measure of heat damage, ranged from 0.91 to 0.95. TMEn values were higher (P = 0.0127) in T-FDN (6.1 kcal/g) and FZN (5.9 kcal/g) than H-FDN (5.3 kcal/g) and were most similar to those estimated by Atwater factors. In general, all diets tested had high AA digestibilities and had TMEn values that were most similar to Atwater factors.

Commercial raw or minimally-processed diets represent a small portion of the pet food market, but the growth of this sector has been significant in recent years. Despite the increasing popularity of commercial frozen and freeze-dried raw diet formats, there is a dearth of literature describing their nutritional properties. The objective of this experiment was to determine the standardized amino acid (AA) digestibilities and nitrogen-corrected true metabolizable energy (TMEn) of raw frozen and freeze-dried dog foods using precision-fed cecectomized and conventional rooster assays. Diets tested included traditional freeze-dried nuggets (T-FDN), frozen nuggets (FZN), and hybrid freeze-dried nuggets (H-FDN). Diets were fed to cecectomized roosters to determine AA digestibilities, while conventional roosters were used to determine TMEn. In both cases, fasted roosters were tube-fed test diets, and excreta was collected. Standardized AA digestibilities were high for all AA (>90% for most indispensable AA) and were not different among diets. The reactive lysine: total lysine ratio, a measure of heat damage, ranged from 0.91 to 0.95. TMEn values were higher in T-FDN (6.1 kcal/g) and FZN (5.9 kcal/g) than H-FDN (5.3 kcal/g). In general, all diets tested had high AA digestibilities and had acceptable reactive lysine:total lysine ratios.

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.

Comparing the standardized amino acid digestibility of an alternative protein source with commercially available protein-based ingredients using the precision-fed cecectomized rooster assay.

Using single-cell-based proteins in pet foods is of interest, but little testing has been done. Therefore, our objective was to determine the amino acid (AA) digestibilities, assess protein quality of a novel microbial protein (MP) (FeedKind), and compare it with other protein-based ingredients using the precision-fed cecectomized rooster assay. Test ingredients included: MP, chicken meal (CM), corn gluten meal (CGM), pea protein (PP), and black soldier fly larvae. Thirty cecectomized roosters (n = 6/ingredient) were randomly assigned to test ingredients. After 24 h of feed withdrawal, roosters were tube-fed 15 g test ingredient and 15 g corn, and then excreta were collected for 48 h. Endogenous AA corrections were made using additional roosters. Digestible indispensable AA score (DIAAS)-like values were calculated to determine protein quality according to Association of American Feed Control Officials (AAFCO), The European Pet Food Industry Federation, and National Research Council reference values for growing and adult dogs and cats. Data were analyzed using the Mixed Models procedure of SAS 9.4, with P ≤ 0.05 being significant. All reactive lysine:total lysine ratios, an indicator of heat damage, were higher than 0.9, except for CM (0.86). Digestibility of indispensable and dispensable AA were >85% and >80% for MP, respectively, with indispensable AA digestibilities being >80% for all other ingredients. In general, CGM had the highest, while CM had the lowest AA digestibilities. Two exceptions were lysine and tryptophan. Lysine digestibility for MP was higher than that of all other ingredients, while tryptophan digestibility for MP was higher than that of CM, CGM, and PP. Threonine digestibility was highest for CGM and MP. Valine digestibility was highest for CGM, PP, and MP. DIAAS-like calculations identified limiting AA of each ingredient and depended on the reference used and life stage and species of animal. Using AAFCO guidelines, all DIAAS-like values for MP were >100 suggesting that it could be used as the sole source of protein in adult dog and cat diets; only methionine had DIAAS-like values <100 for growing kittens. For dogs, limiting AA was most commonly methionine, threonine, and tryptophan in the other protein sources. For cats, limiting AA was most commonly lysine and methionine. Lysine was severely limited in CGM across all life stages considered. Further research in dogs and cats is necessary, but our data suggest that the MP tested has high AA digestibilities and is a high-quality protein source that may be useful in pet foods.

Single-cell-based proteins are of interest for use in pet foods, but little testing has been done. The objective of this experiment was to compare the amino acid (AA) digestibilities and protein quality of a novel microbial protein (MP) (FeedKind) with chicken meal (CM), corn gluten meal (CGM), pea protein (PP), and black soldier fly larvae ingredients using the precision-fed cecectomized rooster assay. Cecectomized roosters were tube-fed the test ingredients and excreta were collected. All reactive lysine:total lysine ratios, an indicator of heat damage, were higher than 0.9, except for CM. Digestibility of indispensable and dispensable AA were >85% and >80% for MP, respectively, with indispensable AA digestibilities being >80% for all other ingredients. In general, CGM had the highest, while CM had the lowest AA digestibilities. Lysine and tryptophan were exceptions, being highest for MP. Threonine and valine digestibilities were also high for MP. Digestible indispensable AA score-like values identified limiting AA of each ingredient. Limiting AA was most commonly methionine, threonine, and tryptophan for dogs and lysine and methionine for cats. Our data suggest that the MP tested has high AA digestibilities and is a high-quality protein source that may be useful in pet foods.

Amino acid digestibility and nitrogen-corrected true metabolizable energy of mildly cooked human-grade vegan dog foods using the precision-fed cecectomized and conventional rooster assays.

The pet food market is constantly changing and adapting to meet the needs and desires of pets and their owners. One trend that has been growing in popularity lately is the feeding of fresh, human-grade foods. Human-grade pet foods contain ingredients that have all been stored, handled, processed, and transported in a manner that complies with regulations set for human food production. While most human-grade pet foods are based on animal-derived ingredients, vegan options also exist. To our knowledge, no in vivo studies have been conducted to analyze the performance of human-grade vegan diets. Therefore, the objective of this study was to investigate the amino acid (AA) digestibility and nitrogen-corrected true metabolizable energy (TME n ) of mildly cooked human-grade vegan dog foods using precision-fed cecectomized rooster and conventional rooster assays. Three commercial dog foods were tested. Two were mildly cooked human-grade vegan dog diets (Bramble Cowbell diet (BC); Bramble roost diet (BR)), while the third was a chicken-based extruded dog diet (chicken and brown rice recipe diet (CT)). Prior to the rooster assays, both mildly cooked diets were lyophilized, and then all three diets were ground. Diets were fed to cecectomized roosters to determine AA digestibility, while conventional roosters were used to determine TME n . All data were analyzed using the mixed models procedure of SAS (version 9.4). The majority of indispensable and dispensable AA across all diets had digestibilities higher than 80%, with a few exceptions (BC: histidine, lysine, threonine, and valine; BR: histidine). The only difference in indispensable AA digestibility among diets was observed with tryptophan, with its digestibility being higher (P = 0.0163) in CT than in BC. TME n values were higher (P = 0.006) in BC and BR (4.55 and 4.66 kcal/g dry matter, respectively) than that in CT (3.99 kcal/g dry matter). The TME n /GE was also higher (P = 0.0193) in BR than in CT. Metabolizable energy (ME) estimates using Atwater factors accurately estimated the energy content of CT, but modified Atwater factors and the predictive equations for ME recommended by the National Research Council underestimated energy content. All calculations underestimated the measured TME n values of BC and BR, with Atwater factors being the closest. Although testing in dogs is required, these data suggest that mildly cooked human-grade vegan dog diets are well-digested. Moreover, TME n data suggest that existing methods and equations underestimate the ME of the mildly cooked human-grade vegan foods tested.

Amino acid digestibility and protein quality of mealworm-based ingredients using the precision-fed cecectomized rooster assay.

Mealworms may serve as an alternative protein source for pet foods because of their high protein content and low environmental footprint. The amino acid (AA) content and protein quality of mealworm-based ingredients may vary depending on their composition and processing, however, so testing is required. Our objective was to measure the AA composition, AA digestibility, and protein quality of mealworm-based ingredients using the precision-fed cecectomized rooster assay. The University of Illinois Institutional Animal Care and Use Committee approved all animal procedures prior to experimentation. Sixteen cecectomized roosters (4 roosters per substrate) were randomly allotted to one of four test substrates: 1) whole lesser mealworm (A. diaperinus) meal (ADw); 2) defatted lesser mealworm (A. diaperinus) meal (ADd); 3) defatted yellow mealworm (T. molitor) meal (TMd); and 4) hydrolyzed T. molitor protein meal (TMh). Ingredients were provided by Ynsect, France. After 26 h of feed withdrawal, roosters were tube-fed test substrates. Following crop intubation, excreta samples were collected for 48 h. Endogenous loss corrections for AA were made by using five additional cecectomized roosters. All data were analyzed using SAS version 9.4. All substrates had high AA digestibilities, with all indispensable AA digestibilities being >90% with the exception of histidine (87.9% to 91.1%) and valine (77.9% to 79.7%). Amino acid digestibilities were not different among substrates (P > 0.05). Digestible indispensable AA score (DIAAS)-like values were calculated to determine protein quality according to Association of American Feed Control Officials (AAFCO) nutrient profiles, The European Pet Food Industry Nutritional Guidelines (FEDIAF) nutritional guidelines, National Research Council (NRC) recommended allowances for adult dogs, adult cats, growing puppies, and growing kittens, and NRC minimal requirements for growing puppies and growing kittens. In general, TMh had the highest and TMd had the lowest DIAAS-like values for most indispensable AA. Methionine (TMh; TMd; ADw) and phenylalanine (ADd) were the first-limiting AA. Our results demonstrate that mealworm-based ingredients are high-quality protein sources. Further research in dogs and cats is necessary to confirm sufficient palatability and digestibility, but these data suggest that they are valuable sources of protein for pet foods.

Mealworms may serve as an alternative protein source for pet foods because of their high protein content and low environmental footprint. The amino acid (AA) content and protein quality of mealworm-based ingredients may vary depending on their composition and processing, however, so testing is required. Our objective was to measure the AA composition, AA digestibility, and protein quality of the following ingredients using the precision-fed cecectomized rooster assay: 1) defatted yellow mealworm (Tenebrio molitor) flour (TMd); 2) hydrolyzed T. molitor proteins (TMh); 3) whole lesser mealworm (Alphitobius diaperinus) flour (ADw); and 4) defatted lesser mealworm (A. diaperinus) flour (ADd). All ingredients had high AA digestibilities, with all indispensable AA digestibilities being >90% with the exception of histidine and valine. Digestible indispensable AA score (DIAAS)-like values were calculated to determine protein quality for adult dogs, adult cats, growing puppies, and growing kittens. In general, TMh had the highest and TMd had the lowest DIAAS-like values for most indispensable AA. Methionine (TMh; TMd; ADw) and phenylalanine (ADd) were the first-limiting AA. Our results demonstrate that mealworm-based ingredients are high-quality protein sources. Dog and cat research is necessary, but these data suggest that they are valuable sources of protein for pet foods.

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.

Amino acid digestibility and digestible indispensable amino acid score-like values of black soldier fly larvae fed different forms and concentrations of calcium using the precision-fed cecectomized rooster assay.

Black soldier fly larvae (BSFL) are an alternative protein source for animals, including dogs and cats. Dietary calcium source is an essential nutrient for BSFL development in the pupal stage. Calcium carbonate (CaCO3) and calcium chloride (CaCl2) are common calcium sources but differ in solubility, acid-binding capacity, and calcium concentration. A high calcium concentration in BSFL may affect how well nitrogen and amino acids (AA) are digested by animals consuming them, thereby affecting feed conversion efficiency. Our objective was to determine the effects of dietary calcium form and concentration on nutrient composition, AA digestibility, and digestible indispensable amino acid score (DIAAS)-like values of BSFL intended for use in animal feeds using the precision-fed cecectomized rooster assay. All BSFL tested in this study were harvested at 18 d after hatch. Industry standard rearing conditions were maintained and a commercial layer ration was fed to all BSFL until 11 d post-hatch. From day 11 to 18, BSFL were fed a combination of distiller's dried grains with solubles from a distillery, bakery byproduct meal, and varied calcium sources. All BSFL diets contained 0.2% calcium in the basal diet plus additional calcium in the following amounts and forms: BSFLA: 1.2% CaCl2, BSFLB: 1.2% CaCO3, BSFLC: 0.75% CaCO3, and BSFLD: 0.6% CaCO3 + 0.6% CaCl2. On day 18, BSFL were washed and frozen. Prior to the rooster assay, BSFL were lyophilized and ground. In total, 16 cecectomized roosters (4 roosters per substrate) were randomly assigned to test substrates. After 24 h of feed withdrawal, roosters were tube-fed 20 g of test substrates. Following crop intubation, excreta were collected for 48 h. Endogenous corrections for AA were made using five additional cecectomized roosters. All data were analyzed using a completely randomized design and the GLM procedure of SAS 9.4. Nutrient and AA digestibilities were not different among substrates. DIAAS-like values were calculated to determine protein quality according to the Association of American Feed Control Officials nutrient profiles and National Research Council recommended allowances for dogs and cats. Although AA digestibilities did not differ, those containing CaCO3 generally had higher DIAAS-like reference values than the diet containing CaCl2 alone (BSFLA). Aromatic AA (Phe + Tyr) and sulfur AA (Met + Cys) were often first-limiting AA. Our results suggest that calcium sources fed to BSFL did not affect AA digestibility and protein quality.

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.

True nutrient and amino acid digestibility of dog foods made with human-grade ingredients using the precision-fed cecectomized rooster assay.

For a pet diet to be labeled as human-grade, every ingredient and the finished food must be stored, handled, processed, and transported according to the current good manufacturing practices for human edible foods. Human-grade dog foods are now available and increasing in popularity, but little research has been conducted to test the digestibility of these foods. For this reason, the objective of this experiment was to determine the true nutrient and amino acid (AA) digestibilities of dog foods formulated with human-grade ingredients using the precision-fed cecectomized rooster assay. Six commercial dog foods were tested, including the Beef & Russet Potato (BRP), Chicken & White Rice (CWR), Fish & Sweet Potato (FSP), Lamb & Brown Rice (LBR), Turkey & Whole Wheat Macaroni (TWM), and Venison & Squash (VSR) formulas provided by Just Food For Dogs LLC (Irvine, CA). Before analysis, all foods were lyophilized and ground. A precision-fed rooster assay using cecectomized roosters was conducted to determine the true nutrient digestibility and standardized AA digestibilities of the foods tested. Conventional roosters were used to determine the nitrogen-corrected true metabolizable energy (TMEn) of the foods. All animal procedures were approved by the University of Illinois Institutional Animal Care and Use Committee prior to experimentation. The substrates and rooster excreta were analyzed for macronutrient and AA composition. All data were analyzed using the Mixed Models procedure of SAS (version 9.4; SAS Institute, Cary, NC). In general, all foods tested were highly digestible. Dry matter digestibility was similar among CWR, LBR, and TWR foods, and greater (P < 0.0001) than that of FSP and VSR foods. Organic matter digestibility was highest (P = 0.0002) for CWR and lowest (P = 0.0002) for VSR. For the majority of indispensable AA, digestibilities were greater than 85%, with some being greater than 90%. TMEn was higher (P < 0.0001) for BRP than the other foods, which were similar to one another. Also, TMEn values were much higher than what would be estimated by using modified Atwater factors and often above the predictive equations for metabolizable energy (ME) recommended by the National Research Council or by using Atwater factors. Although statistical differences were observed among foods, they all performed well and the foods tested had very high AA digestibilities. Additionally, the TMEn data suggest that existing methods and equations for ME prediction underestimate the energy content of the foods tested.

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.

Nutrient and AA digestibility of black soldier fly larvae differing in age using the precision-fed cecectomized rooster assay1.

Edible insects such as black soldier fly larvae (BSFL) are alternative protein sources for animal feeds due to their high-protein content and potential low environmental footprint. However, protein quality and AA content may vary across insect species and age. Our objective was to determine the effects of age on nutrient and AA digestibility of BSFL intended for use in pet foods using the precision-fed cecectomized rooster assay. All animal procedures were approved by the University of Illinois Institutional Animal Care and Use Committee prior to experimentation. Twenty-four cecectomized roosters (four roosters per substrate) were randomly assigned to test substrates [BSFL0 = day 0 (day of hatch); BSFL11 = day 11; BSFL14 = day 14; BSFL18 = day 18; BSFL23 = day 23; BSFL29 = day 29]. After 24 h of feed withdrawal, roosters were tube-fed 20 g of test substrates. Following crop intubation, excreta were collected for 48 h. Endogenous corrections for AA were made using five additional cecectomized roosters. All data were analyzed using a completely randomized design and the GLM procedure of SAS 9.4. DM and OM digestibilities were not different among substrates, but acid-hydrolyzed fat digestibility tended to be greater (P < 0.10) for BSFL23 and BSFL29 than BSFL14 and BSFL18. Although all substrates had a high digestibility, BSFL0 and BSFL11 had the lowest (P < 0.05) digestibilities for most indispensable and dispensable AA. Digestible indispensable AA score (DIAAS)-like values were calculated to determine protein quality according to AAFCO nutrient profiles and NRC recommended allowances for dogs and cats. In general, BSFL18 had the highest, and BSFL11 had the lowest DIAAS-like values for most indispensable AA. Threonine, methionine, and tryptophan were often the first-limiting AA. Our results suggest that BSFL are a high-quality protein and AA source, but that age can affect the AA digestibility and protein quality of this alternative protein source.

Chemical composition, true nutrient digestibility, and true metabolizable energy of chicken-based ingredients differing by processing method using the precision-fed cecectomized rooster assay1.

Chicken-based ingredients are commonly used in pet food products, but vary greatly in nutrient composition and processing conditions that may affect their protein quality and digestibility. Testing the quality of protein sources undergoing different processing conditions provides important information to pet food producers. The objective of this study was to determine the chemical composition, nutrient digestibility, protein, and AA digestibility scores, and nitrogen-corrected true metabolizable energy (TMEn) of chicken-based ingredients that had undergone different processing conditions (i.e., chicken meal, raw chicken, retorted chicken, and steamed chicken) using the precision-fed cecectomized rooster assay. True nutrient digestibility was variable among the protein sources (60% to 76% of DM, 66% to 81% of OM, 83% to 90% of AHF, 50% to 95% of AA and 73% to 85% of TMEn/GE). In general, the chicken meal had a lower (P < 0.05) nutrient digestibility than other ingredients tested, including DM, OM, and most indispensable and dispensable AA, with most having a true digestibility between 75% and 85%. The steamed chicken had the highest indispensable AA digestibilities, with all having a true digestibility greater than 88% and most being over 90%. TMEn value and digestible indispensable AA scores (DIAAS)-like values were higher (P < 0.0001) in the less processed chicken-based ingredients in comparison to chicken meal. Although animal proteins are often considered to be complete proteins, DIAAS-like values <100% suggest that ingredients like chicken meal may not provide all indispensable AA when included at levels to the meet minimal crude protein recommendation. Although raw protein sources are often touted as being the most digestible and of the highest quality, the steamed chicken had the highest (P < 0.0001) DIAAS-like values in this study. This study demonstrates the considerable variability that exists, not only in the chemical composition but also in the true nutrient digestibility among chicken-based ingredients undergoing different processing conditions. These data justify more in vivo testing and the use of DIAAS-like values that consider AA profile, in vivo digestibility, and species-specific recommendations, to evaluate protein-based ingredients intended for use in dog and cat foods.

Development of Intrinsically Labeled Eggs and Poultry Meat for Use in Human Metabolic Research.

BACKGROUND: Stable isotope amino acids are regularly used as tracers to examine whole-body and muscle protein metabolism in humans. To accurately assess in vivo dietary protein digestion and absorption kinetics, the amino acid tracer is required to be incorporated within the dietary protein food source (i.e., intrinsically labeled protein). OBJECTIVE: We assessed the practicality of producing eggs and poultry meat intrinsically labeled with l-[5,5,5-(2)H3]leucine through noninvasive oral tracer administration. METHODS: A specifically formulated diet containing 0.52% leucine was supplemented with 0.3% l-[5,5,5-(2)H3]leucine and subsequently fed to 3 laying hens (Lohmann LSL Whites) for 55 d. On day 55, the hens were slaughtered and their meat, bones, and organs were harvested to determine tissue labeling. In Expt. 1, 2 healthy young men [mean ± SEM age: 22 ± 1.5 y; mean ± SEM body mass index (BMI; in kg/m(2)): 23.7 ± 0.5] ingested 18 g l-[5,5,5-(2)H3]leucine-labeled egg protein. In Expt. 2, 2 healthy young men (mean ± SEM age: 20.0 ± 0.0 y; mean ± SEM BMI: 26.4 ± 3.1) ingested 28 g l-[5,5,5-(2)H3]leucine-labeled poultry meat protein. Plasma samples (Expts. 1 and 2) and muscle biopsies (Expt. 1) were collected before and after labeled-food ingestion. RESULTS: High tracer labeling [>20 mole percent excess (MPE)] in the eggs was obtained after 7 d and maintained throughout the feeding protocol (P < 0.05). Over a 55-d period, ∼850 g egg protein (145 eggs) was produced, with a mean ± SEM tracer enrichment of 22.0 ± 0.8 MPE. Mean ± SEM l-[5,5,5-(2)H3]leucine enrichment in the meat was 9.6 ± 0.1 MPE. In Expts. 1 and 2, the consumption of labeled eggs and poultry meat protein increased plasma l-[5,5,5-(2)H3]leucine enrichment, with mean ± SEM peak values of 6.7 ± 0.1 MPE and 4.0 ± 0.9 MPE, respectively. The mean ± SEM 5-h postprandial increase in myofibrillar l-[5,5,5-(2)H3]leucine enrichment after egg ingestion in healthy young men was 0.051 ± 0.008 MPE (Expt. 1). CONCLUSION: We demonstrated the feasibility of producing intrinsically labeled eggs and poultry meat for use in human metabolic research.