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.

Effects of hydrolyzed chicken liver on digestibility, fecal and urinary characteristics, and fecal metabolites of adult dogs.

High-protein low-carbohydrate diets have been widely used for adult maintenance dogs, as well as in specific weight loss diets and maintenance programs. However, increasing dietary protein may increase undigested protein in the hindgut, modifying intestinal fermentation and fecal metabolite concentrations. The aim of this study was to evaluate the effects of protein source and concentration on apparent total tract digestibility (ATTD) of nutrients, metabolizable energy (ME), fecal and urinary characteristics, and fecal metabolites of dogs. Twelve healthy adult dogs were distributed into six treatments (n = 6 per diet) in a balanced incomplete Latin square design consisting of three periods of 30 days each. Six diets, varying in protein source [poultry byproduct meal (PBPM) and hydrolyzed chicken liver powder (HCLP)] and concentration [24, 32, and 40% crude protein (CP) on dry matter (DM) basis] were tested: PBPM24 (PBPM based diet with 24% CP); PBPM32 (PBPM with 32% CP); PBPM40 (PBPM with 40% CP); HCLP24 (HCLP based diet with 24% CP); HCLP32 (HCLP with 32% CP); HCLP40 (HCLP with 40% CP). The ATTD of CP was greater in dogs-fed HCLP and higher protein concentrations diets (P < 0.05). However, dogs-fed HCLP diets had lower ATTD of fat and carbohydrates, and ME (P < 0.05). Similarly, high-protein diets reduced the ATTD of DM, OM, fat, carbohydrates, and energy (P < 0.05). High-protein diets increased the daily fecal output and moisture (P = 0.004 and P < 0.05, respectively), as well as the fecal score (P < 0.0001), verified as soft, moist stools, but still within the ideal range. Fecal valerate concentration was greater in dogs-fed PBPM at 32% CP (P = 0.007). Fecal isobutyrate tended to increase in dogs-fed PBPM and high-protein diets (P < 0.10). Also, dogs-fed PBPM and high-protein diets had greater fecal concentrations of isovalerate, branched-chain fatty acids (BCFA), and ammonia (P < 0.05). Finally, the fecal lactate concentration increased in dogs-fed HCLP and high-protein diets (P < 0.05). The HCLP increased the ATTD of CP, being a highly digestible protein. Although the inclusion of HCLP slightly increased fecal score and moisture, it decreased the amount of fecal metabolites of protein fermentation ammonia and BCFA, both of which are associated with proteolytic fermentation in the colon.

Feeding companion animals with high-protein diets has been a demand of the market and pet owners. However, the protein quality and quantity consumed can interfere with the amount of undigested protein that reaches the hindgut and be fermented. Intestinal fermentation can be desired when well controlled. This study tested two protein sources (hydrolyzed chicken liver and poultry byproduct meal) combined at three dietary protein concentrations (24, 32, and 40% crude protein on dry matter basis) and their effects on the apparent total tract digestibility (ATTD), fecal and urinary characteristics, and fecal metabolites of healthy adult dogs. In summary, diets containing higher inclusion of hydrolyzed chicken liver had improved protein ATTD. However, the same diets impaired the ATTD of fat and carbohydrates and decreased metabolizable energy. High-protein diets retained more water in the feces and increased the fecal output. Fecal consistency was affected, scored as soft and moist stools, but remained within an acceptable score. Dogs-fed poultry byproduct meal diets had greater concentrations of fecal protein fermentation metabolites, such as ammonia and branched-chain fatty acids, possibly related to a greater amount of undigested protein that reached the hindgut and was fermented.

Effects of Diets Based on Hydrolyzed Chicken Liver and Different Protein Concentrations on the Formation and Deamination of Biogenic Amines and Total Antioxidant Capacity of Dogs.

Biogenic amines are synthesized through the bacterial decarboxylation of amino acids, commonly found in high levels in animal by-product meals due to spoilage. Furthermore, biogenic amines and other metabolites can be produced by the fermentation of proteins in the hindgut according to the protein source and concentration of crude protein (CP) in the diet. Thus, this study aimed to evaluate two protein sources (poultry by-product meal (PBPM) and hydrolyzed chicken liver powder (HCLP)) and three CP concentrations (24, 32, and 40%) and their effects on the consumption and fecal excretion of biogenic amines, plasma monoamine oxidase (MAO) and diamine oxidase (DAO) activities, and total antioxidant capacity (TAC) of healthy adult dogs after 30 days of feeding the experimental diets. Twelve dogs were randomly distributed into six treatments (n = 6/treatment): PBPM24 (PBPM with 24% CP); PBPM32 (PBPM with 32% CP); PBPM40 (PBPM with 40% CP); HCLP24 (HCLP with 24% CP); HCLP32 (HCLP with 32% CP); HCLP40 (HCLP with 40% CP). The PBPM and PBPM-based diets had higher concentrations of putrescine, cadaverine, tyramine, histamine, agmatine, and total biogenic amines. In contrast, HCLP and HCLP-based diets contained higher concentrations of spermidine, phenylethylamine, and spermine. The PBPM and PBPM-diets had higher biogenic amine index (BAI) indicating lower quality due to the high content of putrescine, cadaverine and tyramine. Dogs fed diets with PBPM and higher protein concentrations consumed more putrescine, cadaverine, tyramine, agmatine, and total amines (p < 0.0001), while dogs fed with HCLP consumed more spermidine, phenylethylamine, and spermine (p < 0.0001). Fecal excretion of phenylethylamine was greater in dogs fed HCLP32 and HCLP40 diets (p = 0.045). Dogs fed with HCLP tended to excrete more spermidine and tryptamine via feces, while higher protein concentrations tended to increase fecal excretion of cadaverine (p < 0.10). Plasma MAO activity was higher in dogs fed HCLP24 and PBPM32 diets (p = 0.024). The plasma activities of DAO and TAC were not different between diets (p > 0.05). Although we did not evaluate the intestinal activities of MAO and DAO, our results suggest that healthy adult dogs have an efficient deamination process on the gut epithelium.

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.

Hydrolyzed chicken liver used as single source of animal protein in diet and its effect on cytokines, immunoglobulins, and fecal microbiota profile of adult dogs.

Dogs with food allergies and enteropathies may require hydrolyzed diets to prevent or reduce clinical signs, therefore the protein sources used in these diets must be previously characterized and evaluated in healthy dogs. This study aimed to investigate the effects of a hydrolyzed chicken liver powder-based diet (HCLP) versus a poultry by-product meal and bovine meat and bone meal-based diet (Control), on complete blood count (CBC), cytokine, immunoglobulins responses (assessed on days 0, 15, 30 and 45), and fecal microbiota (assessed on day 45) in healthy adult dogs. The CBC did not differ between diets (P>0.05), remaining within reference range. Total plasma IL-4 concentrations were decreased over time independent of the dietary treatment (P<0.001). Total plasma IgA decreased on day 30 compared to days 0 and 45 in dogs fed the control diet (P<0.001). Total plasma IgE concentrations were reduced on days 30 and 45 in dogs fed the control diet, and on days 15 vs 30 and 15 vs 45 in dogs fed HCLP diet (P = 0.001). The 16S rRNA gene sequencing showed similar species richness and abundances of phyla and genera between diets (P>0.05). ß-diversity principal coordinate analysis plots demonstrated that HCLP group had a higher similarity than control. Based on our results, healthy adult dogs fed a HCLP based diet maintained normal values for hematological and immunological characteristics, and fecal microbiota after 45 days of feeding.

Determination of endogenous fat loss and true total tract digestibility of fat in mink (Neovison vison).

The objective of this study was to determine the endogenous fat loss (EFL) and to calculate true total tract digestibility (TTTD) of fat in mink (Neovison vison) using soybean oil-based diets with different fat levels. In the digestibility assay, four diets with 6.30%, 13.9%, 22.0% and 34.0% fat in dry matter were used. Sixteen adult male mink were distributed in a complete randomised design. The apparent total tract digestibility (ATTD) of dietary fat was 90.8%, 95.9%, 96.9% and 97.8%, respectively. The apparent total digestible fat was linearly related to dietary fat intake (r2 = 0.99). The EFL was estimated from the slope of the regression equation and was determined to be 5.09 g/kg DM intake. The TTTD of soybean oil was determined to be 99.3%. Therefore, TTTD values will have negligible impact in feed formulation as they are close to ATTD values with the dietary fat levels normally used for mink.

Soybean oil and beef tallow in dry extruded diets for adult dogs.

The aim of the study was to determine the effects of two different fat sources (soybean oil (SO) and beef tallow (BT)) in dry extruded dog diets on the intake of food and metabolizable energy (ME), on faecal characteristics and apparent total tract digestibility (ATTD) of nutrients and energy. Ten adult dogs of different breeds were used in a Latin square design. Five experimental diets were designed from a basal diet. A Control diet was coated with 1% SO and four other diets were obtained by coating the basal diet with 6.5% and 13% of SO or BT. The Control, 6.5% and 13% coated diets contained approximately 8.5%, 15% and 20% fat, respectively. The dogs had similar dry matter (DM) intakes and, consequently, higher ME intakes with an increased fat content for both sources (p < 0.05). Fat digestibility was highest for Diet SO13% (p < 0.05). The ATTD of DM and organic matter was highest (p < 0.05) for Diets SO13% and BT13%. Fat coating improved the faecal score, especially for Diet BT13%. The dietary ratios of protein:fat:carbohydrate [% of ME] were close to AAFCO's estimates. For SO a higher ATTD (99.1%) and ME content (38.88 MJ/kg) was estimated than for BT (ATTD 92.9% and 36.37 MJ ME/kg). Both SO and BT can comprise up to 13% of the diet. However, SO was more susceptible to leaking from the kibbles with the coating method applied.

Produtividade e valor nutritivo de pastos consorciados com diferentes espécies de leguminosas / Productivity and nutritive value of mixed pastures with different legume species

O objetivo desta pesquisa foi avaliar a produtividade e o valor nutritivo de dois sistemas forrageiros (SF) constituídos por capim elefante (CE), azevém (AZ), trevo branco (TB) e espécies de crescimento espontâneo (ECE), como SF1; e CE + AZ + amendoim forrageiro (AF) + ECE, como SF2. O CE foi estabelecido em linhas espaçadas a cada quatro metros. Entre elas, foi estabelecido azevém, durante o período hibernal, permitindo-se o desenvolvimento de ECE no período estival. Para avaliação, foram utilizadas vacas da raça Holandesa e o método de pastejo foi o rotacionado. Avaliaram-se a massa de forragem inicial (MFI) e a composição botânica e estrutural dos pastos. Para as estimativas de valor nutritivo, foram feitas amostragens simulando o pastejo e analisados a fibra em detergente neutro (FDN), fibra em detergente ácido (FDA), proteína bruta (PB), digestibilidade in vitro da matéria seca (DIVMS) e digestibilidade in vitro da matéria orgânica (DIVMO). O delineamento experimental foi o inteiramente casualizado com dois tratamentos e duas repetições, em parcelas subdivididas no tempo. Os valores médios de massa de forragem inicial e da carga animal foram similares entre os SF. Observou-se resultado superior para o teor médio de PB da forragem do SF2. O uso do azevém, das leguminosas e das espécies de crescimento espontâneo em consórcio com o CE permite manter massa de forragem uniforme no decorrer dos pastejos.

The objective of this research was to evaluate the productivity and nutritive value of two pasture-based systems (PS) with elephant grass (EG) + ryegrass (RG) + white clover (WC) + spontaneous growing species (SGS), for PS1; and EG + RG + forage peanut (FP) + SGS, for PS2. EG was established in rows spaced by four meters. Ryegrass was seeding in the space between rows during cool-season and development of SGS was permitted in the warm-season. Holstein cows under rotational grazing were used in the evaluation. The initial forage mass (IFM), the botanical composition and the structural components were evaluated. Samples from hand-plucking method were collected to analyze neutral detergent fiber (NDF), acid detergent fiber (ADF), crude protein (CP), in vitro dry matter digestibility (IVDMD) and in vitro organic matter digestibility (IVOMD). The experimental design used was completely randomized with two treatments, two repetitions in complete split-plot time. The mean values of initial forage mass and stocking rate were similar among PS. Superior results were found for mean of CP in PS2. The use of ryegrass, legumes and spontaneous growing species mixed to EG allow the forage mass to be uniform during the grazing pastures.

Fracionamento dos carboidratos de silagens de milho safrinha colhidas em diferentes alturas de corte / Carbohydrates fractioning of late crop corn silages harvested at different cutting height

Este trabalho teve como objetivo verificar se há alteração na composição dos carboidratos de silagens de milho safrinha quando colhidas nas alturas de corte (AC) de 20, 45, 70 e 95cm acima do solo. O experimento foi conduzido em delineamento completamente casualizado, com quatro unidades experimentais (minisilos) por tratamento. A matéria seca aumentou linearmente (MS=27,25+0,06928*AC, onde: R²=0,82 e P<0,01) com a elevação da AC pela maior proporção de grãos e menor proporção de colmo na massa ensilada. Os valores de fibra em detergente neutro corrigida para cinzas e proteína (FDNcp) e lignina (LDA) diminuíram com a elevação na AC, FDNcp=43,75-0,08411*AC (R²=0,47 e P<0,01) e LDA=6,983-0,03156*AC (R²=0,49 e P<0,01), respectivamente. O teor de carboidratos não-fibrosos (CNF=41,57+0,07861*AC, onde: R²=0,54 e P<0,01) e a fração B1 dos carboidratos (B1=27,16+0,1239*AC, em que: R²=0,66 e P<0,01) aumentaram com a elevação da AC. O cultivo de milho durante a época denominada safrinha na região Sul do Brasil proporciona a produção de silagem de alta qualidade por apresentar alto teor de amido e baixo de fibra em detergente neutro. A elevação na altura de corte das plantas de milho altera a distribuição dos carboidratos aumentando a proporção de amido e reduzindo o teor de fibras e lignina na silagem.

The objective was to verify if there are changes in the carbohydrates composition of late crop corn silages harvested at 20, 45, 70 and 95cm of cutting height (CH). The research was conducted in a randomized entirely design with four replications (mini-silos) per treatment. The dry matter content increased linearly DM=27.25+0.06928*CH (R²=0.82 and P<0.01) with the CH elevation due to the larger grain proportion in the ensiled mass. The neutral detergent fiber corrected for ashes and protein (NDFap) and lignin (ADL) values lowered with the CH elevation, NDFap=43.75-0.08411*CH (R²=0.47 and P<0.01) and ADL=6.983-0.03156*CH (R²=0.49 and P<0.01), respectively. The carbohydrates B1=27.16+0.1239*CH (R²=0.66 and P<0.01) fraction and the non-fiber carbohydrates NFC=41.57+0.07861*CH (R²=0.54 and P<0.01) increased linearly with the CH elevation. The late corn cropping in the south region of Brazil lead to high quality silages, with high starch and low neutral detergent fiber values. The CH elevation in the corn plant changes the carbohydrates distribution, with higher starch and lower neutral detergent fiber and lignin values in the silage.