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.

Inulin alleviates perinatal 2-ethylhexyl diphenyl phosphate (EHDPHP) exposure-induced intestinal toxicity by reshaping the gut microbiota and suppressing the enteric-origin LPS/TLR4/NF-κb pathway in dams and pups.

Organophosphorus flame retardants (OPFRs), such as 2-ethylhexyl diphenyl phosphate (EHDPHP), are ubiquitously used, leading to pervasive environmental contamination and human health risks. While associations between EHDPHP and health issues such as disruption of hormones, neurotoxic effects, and toxicity to reproduction have been recognized, exposure to EHDPHP during perinatal life and its implications for the intestinal health of dams and their pups have largely been unexplored. This study investigated the intestinal toxicity of EHDPHP and the potential for which inulin was effective. Dams were administered either an EHDPHP solution or a corn oil control from gestation day 7 (GD7) to postnatal day 21 (PND21), with inulin provided in their drinking water. Our results indicate that inulin supplementation mitigates damage to the intestinal epithelium caused by EHDPHP, restores mucus-secreting cells, suppresses intestinal hyperpermeability, and abates intestinal inflammation by curtailing lipopolysaccharide leakage through reshaping of the gut microbiota. A reduction in LPS levels concurrently inhibited the inflammation-associated TLR4/NF-κB pathway. In conclusion, inulin administration may ameliorate intestinal toxicity caused by EHDPHP in dams and pups by reshaping the gut microbiota and suppressing the LPS/TLR4/NF-κB pathway. These findings underscore the efficacy of inulin as a therapeutic agent for managing health risks linked to EHDPHP exposure.

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.

Gut microbiota-derived short-chain fatty acids ameliorate methamphetamine-induced depression- and anxiety-like behaviors in a Sigmar-1 receptor-dependent manner.

Methamphetamine (Meth) abuse can cause serious mental disorders, including anxiety and depression. The gut microbiota is a crucial contributor to maintaining host mental health. Here, we aim to investigate if microbiota participate in Meth-induced mental disorders, and the potential mechanisms involved. Here, 15 mg/kg Meth resulted in anxiety- and depression-like behaviors of mice successfully and suppressed the Sigma-1 receptor (SIGMAR1)/BDNF/TRKB pathway in the hippocampus. Meanwhile, Meth impaired gut homeostasis by arousing the Toll-like receptor 4 (TLR4)-related colonic inflammation, disturbing the gut microbiome and reducing the microbiota-derived short-chain fatty acids (SCFAs). Moreover, fecal microbiota from Meth-administrated mice mediated the colonic inflammation and reproduced anxiety- and depression-like behaviors in recipients. Further, SCFAs supplementation optimized Meth-induced microbial dysbiosis, ameliorated colonic inflammation, and repressed anxiety- and depression-like behaviors. Finally, Sigmar1 knockout (Sigmar1-/-) repressed the BDNF/TRKB pathway and produced similar behavioral phenotypes with Meth exposure, and eliminated the anti-anxiety and -depression effects of SCFAs. The activation of SIGMAR1 with fluvoxamine attenuated Meth-induced anxiety- and depression-like behaviors. Our findings indicated that gut microbiota-derived SCFAs could optimize gut homeostasis, and ameliorate Meth-induced mental disorders in a SIGMAR1-dependent manner. This study confirms the crucial role of microbiota in Meth-related mental disorders and provides a potential preemptive therapy.

Multi-Omics Analysis Reveals the Role of Sigma-1 Receptor in a Takotsubo-like Cardiomyopathy Model.

Takotsubo syndrome (TTS) is a stress-induced cardiomyopathy that presents with sudden onset of chest pain and dyspneic and cardiac dysfunction as a result of extreme physical or emotional stress. The sigma-1 receptor (Sigmar1) is a ligand-dependent molecular chaperone that is postulated to be involved in various processes related to cardiovascular disease. However, the role of Sigmar1 in TTS remains unresolved. In this study, we established a mouse model of TTS using wild-type and Sigmar1 knockout mice to investigate the involvement of Sigmar1 in TTS development. Our results revealed that Sigmar1 knockout exacerbated cardiac dysfunction, with a noticeable decrease in ejection fraction (EF) and fractional shortening (FS) compared to the wild-type model. In terms of the gut microbiome, we observed regulation of Firmicutes and Bacteroidetes ratios; suppression of probiotic Lactobacillus growth; and a rise in pathogenic bacterial species, such as Colidextribacter. Metabolomic and transcriptomic analyses further suggested that Sigmar1 plays a role in regulating tryptophan metabolism and several signaling pathways, including MAPK, HIF-1, calcium signaling, and apoptosis pathways, which may be crucial in TTS pathogenesis. These findings offer valuable insight into the function of Sigmar1 in TTS, and this receptor may represent a promising therapeutic target for TTS.

Inulin alleviates neuroinflammation and oxidative stress induced by perinatal 2-ethylhexyl diphenyl phosphate (EHDPHP) exposure in female mice and offspring.

Organophosphorus flame retardants (OPFRs), including 2-ethylhexyl diphenyl phosphate (EHDPHP), are prevalent in everyday life due to their broad usage in fields such as healthcare, electronics, industry, and sports. These compounds, added to polymers through physical mixing, can leach into the environment, posing a risk to humans through direct contact or the food chain. Despite known associations with health issues like endocrine disruption, neurotoxicity, and reproductive toxicity, the implications of perinatal EHDPHP exposure on both mothers and offspring are still unclear. This study aimed to investigate the neuroinflammatory effects of EHDPHP and the potential mitigating role of inulin. Pregnant C57 mice were administered either a corn oil control or an EHDPHP solution (300 µg/kg bw/d) from gestation day 7 (GD7) to postnatal day 21 (PND21). Concurrently, mice were provided either regular drinking water or water supplemented with 1% inulin. We found that EHDPHP significantly increased the serum levels of IL-1ß, IL-6, and MDA, but decreased SOD levels in both mothers and pups. These effects were reversed by inulin supplementation. RNA-sequencing revealed that EHDPHP induced inflammation and oxidative stress through the TLR4/NF-κB pathway, which was mitigated by inulin. In conclusion, inulin ameliorated EHDPHP-induced neuroinflammation and oxidative stress in both mothers and offspring, highlighting its potential therapeutic role.

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.

Epigallocatechin-3-gallate ameliorates polystyrene microplastics-induced anxiety-like behavior in mice by modulating gut microbe homeostasis.

Polystyrene microplastics (PS-MPs) have emerged as a concerning pollutant in modern society due to their widespread production and usage. Despite ongoing research efforts, the impact of PS-MPs on mammalian behavior and the mechanisms driving these effects remain incompletely elucidated. Consequently, effective strategies for prevention have yet to be developed. To fill these gaps, C57BL/6 mice were orally administered with 5 µm PS-MPs for 28 consecutive days in this study. The open-field test and the elevated plus-maze test were performed to evaluate the anxiety-like behavior, 16S rRNA sequencing and untargeted metabolomics analysis were used to detect the changes of gut microbiota and serum metabolites. Our results indicated that PS-MPs exposure activated hippocampal inflammation and induced anxiety-like behavior in mice. Meanwhile, PS-MPs disturbed the gut microbiota, impaired the intestinal barrier, and aroused peripheral inflammation. Specifically, PS-MPs increased the abundance of pathogenic microbiota Tuzzerella, while lowered the abundance of probiotics Faecalibaculum and Akkermansia. Interestingly, eliminating the gut microbiota protected against the deleterious effects of PS-MPs on intestinal barrier integrity, reduced the levels of peripheral inflammatory cytokines, and ameliorated anxiety-like behavior. Additionally, green tea's primary bioactive constituent, epigallocatechin-3-gallate (EGCG), optimized gut microbial composition, improved intestinal barrier function, reduced peripheral inflammation, and exerted anti-anxiety effects by inhibiting the hippocampal TLR4/MyD88/NF-κB signaling cascade. EGCG also remodeled serum metabolism, especially modulated purine metabolism. These findings suggested that gut microbiota participates in PS-MPs-induced anxiety-like behavior by modulating the gut-brain axis, and that EGCG could serve as a potential preventive strategy.

Microbial community succession in the intestine of mice with deep partial-thickness burns.

Introduction: Burn injury has been shown to lead to changes in the composition of the gut microbiome and cause other damage in patients. However, little is known about how the gut microbial community evolves in individuals who have recovered from burn injury. Methods: In this study, we established a model of deep partial-thickness burn in mice and collected fecal samples at eight time points (pre-burn, 1, 3, 5, 7, 14, 21, and 28 days post-burn) for 16S rRNA amplification and high-throughput sequencing. Results: The results of the sequencing were analyzed using measures of alpha diversity, and beta diversity and taxonomy. We observed that the richness of the gut microbiome declined from day 7 post-burn and that the principal component and microbial community structure varied over time. On day 28 after the burn, the microbiome composition largely returned to the pre-burn level, although day 5 was a turning point for change. Some probiotics, such as the Lachnospiraceae_NK4A136_group, decreased in composition after the burn but were restored in the later recovery period. In contrast, Proteobacteria showed an opposite trend, which is known to include potential pathogenic bacteria. Conclusion: These findings demonstrate gut microbial dysbiosis after burn injury and provide new insights into the burn-related dysbiosis of the gut microbiome and strategies for improving the treatment of burn injury from the perspective of the microbiota.