Effect of feeding frequency on reproductive performances and stress responses in gestating sows.

The objective of this study was to investigate the influence of feeding frequency on a sow's reproductive performance and stress response during gestation. A total of twenty multiparous sows (Yorkshire × Landrace) were used in a completely randomized design based on their parity, body weight (BW), and backfat thickness (BFT), and the sows were allotted to two different feeding systems: 1) once daily feeding (OF) and 2) twice daily feeding (TF) in corn-soybean meal based diets. The gestation diet was formulated to contain 3,265 kcal of metabolizable energy (ME) / kg, 12.90% of crude protein (CP), and 0.75 % of total lysine. The lactation diet was formulated to contain 3,265 kcal of ME / kg, 16.80% of CP, and 1.08% of total lysine and provided ad libitum during lactation. In gestation, sow BFT and BF changes were not affected by feeding frequency, but higher BW and BW gain from day 35 to 90 and day 35 to 110 were observed in OF sow (p < 0.10). In lactation, feeding frequency did not influence on BW, BW gain, BFT, BF changes, average daily feed intake, and wean-to-estrus interval. Also, there were no differences in litter size, litter weight and piglet weight in lactating sows. OF sows had higher (p < 0.05; p < 0.10) protein, solid-not-fat, and total solid concentrations in colostrum compared to TF sows, while OF sows had a lower (p < 0.05) lactose concentration in colostrum compared to TF sows. Sows in OF showed significantly lower average daily water consumption (ADWC) from day 35 to 110 of gestation (p < 0.05). While there were no significant differences in stereotypic behaviors and salivary cortisol levels during gestation between treatments, the OF sows showed less time spending on the activity at day 105 (p < 0.05). In conclusion, reduced feeding frequency increased BW gain during gestation, decreased activation time, and changed the colostrum composition. This information may contribute to the understanding of the physiological and behavioral change of gestating sows by manipulating feeding frequency.

The Effect of Reduced Crude Protein on Growth Performance, Nutrient Digestibility, and Meat Quality in Weaning to Finishing Pigs.

This study aimed to evaluate optimal crude protein (CP) levels based on the National Research Council guidelines from 1998 and 2012 and their impacts on growth performance, fecal score, blood profiles, nutrient digestibility, and carcass characteristics of weaning to finishing pigs. Four diets were established in this experiment: high-protein (HP), medium-high-protein (MHP), medium-low-protein (MLP), and low-protein (LP). The HP diet followed the NRC (1998) guidelines, while the MHP diet reduced the CP content by 1%. The MLP diet had 1.5% lower CP content than the HP diet. The LP diet followed the NRC guideline of 2012, which suggests less protein than the NRC guideline of 1998. There were no significant differences in body weight, average daily feed intake, or nutrient digestibility. However, the average daily gain (ADG) of pigs fed the LP diet at 7-10 weeks was lower than in the other treatments, whereas the ADG of the pigs fed the LP diet was higher compared with that of pigs fed the other diets, showing compensatory growth in finishing periods (p < 0.05). The blood urea nitrogen of pigs fed the LP diet showed the lowest value, whereas the highest value was found in pigs fed the HP diet, and the other two diets were similar to the HP diet or positioned between the HP and LP diets (p < 0.05). Water holding capacity, cooking loss, shear force, and pH in the longissimus muscle were not influenced by varying dietary CP levels (p > 0.05), but the hunter values L and b were increased in pigs fed the LP diet (p < 0.05). Conclusively, a low-protein diet did not negatively affect growth performance, nutrient digestibility, or meat quality.

RNA-mediated demixing transition of low-density condensates.

Biomolecular condensates play a key role in organizing cellular reactions by concentrating a specific set of biomolecules. However, whether condensate formation is accompanied by an increase in the total mass concentration within condensates or by the demixing of already highly crowded intracellular components remains elusive. Here, using refractive index imaging, we quantify the mass density of several condensates, including nucleoli, heterochromatin, nuclear speckles, and stress granules. Surprisingly, the latter two condensates exhibit low densities with a total mass concentration similar to the surrounding cyto- or nucleoplasm. Low-density condensates display higher permeability to cellular protein probes. We find that RNA tunes the biomolecular density of condensates. Moreover, intracellular structures such as mitochondria heavily influence the way phase separation proceeds, impacting the localization, morphology, and growth of condensates. These findings favor a model where segregative phase separation driven by non-associative or repulsive molecular interactions together with RNA-mediated selective association of specific components can give rise to low-density condensates in the crowded cellular environment.

The Role of Dietary Fiber in Improving Pig Welfare.

This review aims to discuss the effects of dietary fiber sources with various levels on stereotypic behaviors in sows. There are a variety of dietary fiber sources that are supplemented to feeds for sows. However, dietary fiber sources have different physio-chemical properties, leading to controversial results in feed motivation, nutrient digestibility, and behaviors in sows fed fiber-rich diets. Findings from previous studies indicated that soluble fiber delays nutrient absorption and decreases physical activity after feeding. In addition to this, it increases volatile fatty acid production, provides energy, and prolongs the feeling of satiety. It also prevents certain stereotypies and thus is paramount to sow welfare.

Dietary enrichment of resistant starches or fibers differentially alter the feline fecal microbiome and metabolite profile.

BACKGROUND: Cats are strict carnivores but possess a complex gastrointestinal (GI) microbial community that actively ferments dietary substrates that are not digested and reach the colon. The GI microbiota responses to dietary inclusion of resistant starches versus fibers have not been tested in cats. Thus, our objective was to evaluate the effects of diets enriched in resistant starch or fibers on the fecal characteristics, microbiome, and metabolite profiles of cats. Twelve healthy adult domestic shorthair cats (age = 9.6 ± 4.0 year; body weight = 3.9 ± 1.0 kg) were used in a replicated 3 × 3 Latin square design to test diets that were enriched with: (1) resistant starch (ERS), (2) a fiber-prebiotic-probiotic blend (FPPB), or (3) a fiber-prebiotic-probiotic blend + immune-modulating ingredients (iFPPB). In each 28-day period, 22 days of diet adaptation was followed by fecal and blood sample collection. Fecal samples were used for shotgun metagenomic sequencing. In addition, fecal and blood metabolite measurements and white blood cell stimulation was performed to assess immune function. RESULTS: A total of 1690 bacterial species were identified, with 259 species differing between fiber-rich and ERS treatments. In comparison with fiber-rich treatments that increased diversity and promoted Firmicutes and Bacteroidetes populations, resistant starch reduced microbial diversity and fecal pH, led to a bloom in Actinobacteria, and modified Kyoto Encyclopedia of Genes and Genomes orthology (KO) terms pertaining to starch and sucrose metabolism, fatty acid biosynthesis and metabolism, epithelial cell signaling, among others. Resistant starch also differentially modified fecal metabolite concentrations with relevance to GI and overall host health (increased butyrate; decreased propionate and protein catabolites - branched-chain fatty acids; phenols and indoles; ammonia) and reduced blood cholesterol, which correlated strongly with microbial taxa and KO terms, and allowed for a high predictive efficiency of diet groups by random forest analysis. CONCLUSION: Even though domestic cats and other carnivores evolved by eating low-carbohydrate diets rich in protein and fat, our results demonstrate that the feline microbiome and metabolite profiles are highly responsive to dietary change and in directions that are predictable.

Engineering DNA-based synthetic condensates with programmable material properties, compositions, and functionalities.

Biomolecular condensates participate in diverse cellular processes, ranging from gene regulation to stress survival. Bottom-up engineering of synthetic condensates advances our understanding of the organizing principle of condensates. It also enables the synthesis of artificial systems with novel functions. However, building synthetic condensates with a predictable organization and function remains challenging. Here, we use DNA as a building block to create synthetic condensates that are assembled through phase separation. The programmability of intermolecular interactions between DNA molecules enables the control over various condensate properties including assembly, composition, and function. Similar to the way intracellular condensates are organized, DNA clients are selectively partitioned into cognate condensates. We demonstrate that the synthetic condensates can accelerate DNA strand displacement reactions and logic gate operation by concentrating specific reaction components. We envision that the DNA-based condensates could help the realization of the high-order functions required to build more life-like artificial systems.

Formation of non-base-pairing DNA microgels using directed phase transition of amphiphilic monomers.

Programmability of DNA sequences enables the formation of synthetic DNA nanostructures and their macromolecular assemblies such as DNA hydrogels. The base pair-level interaction of DNA is a foundational and powerful mechanism to build DNA structures at the nanoscale; however, its temperature sensitivity and weak interaction force remain a barrier for the facile and scalable assembly of DNA structures toward higher-order structures. We conducted this study to provide an alternative, non-base-pairing approach to connect nanoscale DNA units to yield micrometer-sized gels based on the sequential phase transition of amphiphilic unit structures. Strong electrostatic interactions between DNA nanostructures and polyelectrolyte spermines led to the formation of giant phase-separated aggregates of monomer units. Gelation could be initiated by the addition of NaCl, which weakened the electrostatic DNA-spermine interaction while attractive interactions between cholesterols created stable networks by crosslinking DNA monomers. In contrast to the conventional DNA gelation techniques, our system used solid aggregates as a precursor for DNA microgels. Therefore, in situ gelation could be achieved by depositing aggregates on the desired substrate and subsequently initiating a phase transition. Our approach can expand the utility and functionality of DNA hydrogels by using more complex nucleic acid assemblies as unit structures and combining the technique with top-down microfabrication methods.

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

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

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

Dietary supplementation with fiber, "biotics," and spray-dried plasma affects apparent total tract macronutrient digestibility and the fecal characteristics, fecal microbiota, and immune function of adult dogs.

A variety of functional ingredients, including fibers, prebiotics, probiotics, and postbiotics may be added to pet foods to support gastrointestinal and immune health. While many of these ingredients have been tested individually, commercial foods often include blends that also require testing. This study was conducted to evaluate the effects of diets containing blends of fibers, "biotics," and/or spray-dried plasma on apparent total tract digestibility (ATTD), stool quality, fecal microbiota and metabolites, and immune health outcomes of adult dogs. A total of 12 healthy adult intact English pointer dogs (6 M, 6 F; age = 6.4 ± 2.0 yr; BW = 25.8 ± 2.6 kg) were used in a replicated 3 × 3 Latin square design to test diets formulated to: 1) contain a low concentration of fermentative substances (control diet, CT); 2) be enriched with a fiber-prebiotic-probiotic blend (FPPB); and 3) be enriched with a fiber-prebiotic-probiotic blend + immune-modulating ingredients (iFFPB). In each 28-d period, 22 d of diet adaptation was followed by a 5-d fecal collection phase and 1 d for blood sample collection. All data were analyzed using SAS 9.4, with significance being P < 0.05 and trends being P < 0.10. FPPB and iFPPB diets led to shifts in numerous outcome measures. Dry matter (DM), organic matter, fat, fiber, and energy ATTD were lower (P < 0.01), fecal scores were lower (P < 0.01; firmer stools), and fecal DM% was higher (P < 0.0001) in dogs fed FPPB or iFPPB than those fed CT. Serum triglycerides and cholesterol were lower (P < 0.01) in dogs fed FPPB or iFPPB than those fed CT. Fecal protein catabolites (isobutyrate, isovalerate, indole, and ammonia) and butyrate were lower (P < 0.05), while fecal immunoglobulin A (IgA) was higher (P < 0.01) in dogs fed FPPB and iFPPB than those fed CT. Fecal microbiota populations were affected by diet, with alpha-diversity being lower (P < 0.05) in dogs fed iFPPB and the relative abundance of 20 bacterial genera being altered in dogs fed FPPB or iFPPB compared with CT. The circulating helper T cell:cytotoxic T cell ratio was higher (P < 0.05) in dogs fed iFPPB than those fed CT. Circulating B cells were lower (P < 0.05) in dogs fed FPPB than those fed iFPPB, and lower (P < 0.05) in dogs fed iFPPB than those fed CT. Our results demonstrate that feeding a fiber-prebiotic-probiotic blend may provide many benefits to canine health, including improved stool quality, beneficial shifts to fecal microbiota and metabolite profiles, reduced blood lipids, and increased fecal IgA.

A variety of functional ingredients­those that provide benefits beyond their nutritional value­may be added to pet foods to support gastrointestinal and immune health. While many of these ingredients have been tested individually, commercial foods often include blends that also require testing. This study was conducted to evaluate the effects of diets containing blends of dietary fibers and other functional ingredients on nutrient digestibility and the stool characteristics and immune health outcomes of adult dogs consuming them. Treatments included a control diet containing low amounts of dietary fiber, a diet containing a fiber­prebiotic­probiotic blend, and a diet containing the fiber­prebiotic­probiotic blend as well as immune-modulating ingredients. The test diets were shown to shift many outcome measures. First, they were shown to reduce nutrient digestibility and decrease fecal scores (more firm stool). Second, test diets reduced blood lipids and beneficially altered fecal metabolite concentrations. Third, test diets increased fecal immunoglobulin A concentrations, suggesting enhanced gut immunity. Lastly, the test diets shifted fecal bacterial populations. Our results demonstrate that feeding a fiber­prebiotic­probiotic blend may provide many benefits to canine health, including improved stool quality, beneficial shifts to fecal bacteria and metabolite profiles, reduced blood lipids, and enhanced gut immunity.

The flexibility-based modulation of DNA nanostar phase separation.

Phase separation of biomolecules plays key roles in physiological compartmentalization as well as pathological aggregation. A deeper understanding of biomolecular phase separation requires dissection of a relation between intermolecular interactions and resulting phase behaviors. DNA nanostars, multivalent DNA assemblies of which sticky ends define attractive interactions, represent an ideal system to probe this fundamental relation governing phase separation processes. Here, we use DNA nanostars to systematically study how structural flexibility exhibited by interacting species impacts their phase behaviors. We design multiple nanostars with a varying degree of flexibility using single-stranded gaps of different lengths in the arm of each nanostar unit. We find that structural flexibility drastically alters the phase diagram of DNA nanostars in such a way that the phase separation of more flexible structures is strongly inhibited. This result is not due to self-inhibition from the loss of valency but rather ascribed to a generic flexibility-driven change in the thermodynamics of the system. Our work provides not only potential regulatory mechanisms cells may exploit to dynamically control intracellular phase separation but also a route to build synthetic systems of which assembly can be controlled in a signal dependent manner.

Effects of Lysine Cell Mass Supplementation as a Substitute for L-Lysine·HCl on Growth Performance, Diarrhea Incidence, and Blood Profiles in Weaning Pigs.

This study was conducted to evaluate the effects of lysine cell mass (LCM) as an alternative lysine source in diets for weaning pigs on growth performance, diarrhea incidence, and blood profiles. In experiment 1, a total of 200 weaning pigs, with an average body weight (BW) of 6.89 ± 1.04 kg, were allotted into one of five treatments with four replicates of 10 pigs per pen in a randomized complete block design (RCBD). The dietary treatments were composed of LCM supplementation (0, 0.25, 0.5, 0.75, or 1.0%) with partial replacement of L-lysine·HCl (0 to 0.8% for phase 1 diets and 0 to 0.07% for phase 2 diets). The BW and feed intake were recorded at the end of each phase (d 0 to 14 for phase 1, d 14 to 35 for phase 2), and diarrhea incidence was checked daily throughout the experimental period. Blood samples were taken from the jugular vein of pigs at 2 weeks and 5 weeks to determine the blood profiles of weaning pigs. In experiment 2, a total of 144 weaning pigs with an average BW of 6.44 ± 1.19 kg were allotted into one of six treatments with six replicates of four pigs per pen in RCBD. The dietary treatments were composed of LCM supplementation (0 to 3.5% for phase 1 diets and 0 to 2.2% for phase 2 diets) with replacement of L-lysine·HCl from 0 to 100%. In experiment 1, partial replacement of L-lysine·HCl with 0 to 1% LCM did not affect growth performance and diarrhea incidence of pigs. An increase in the LCM supplementation from 0 to 1% with partial replacement of L-lysine·HCl had no influence on the blood urea nitrogen concentrations, whereas it resulted in a linear decrease (p < 0.05) in the serum IgG concentrations for 5 weeks. In experiment 2, increasing the dietary level of LCM with replacement of L-lysine·HCl quadratically decreased (p < 0.05) ADG and G-F ratio for phase 2 and G-F ratio for the overall period such that 100% replacement of L-lysine·HCl with LCM decreased ADG and G-F ratio of weaning pigs. An increase in the LCM supplementation with replacement of L-lysine·HCl tended to decrease linearly (p < 0.10) the diarrhea incidence of weaning pigs for the overall period and linearly decrease (p < 0.05) the serum IgG concentrations for 2 weeks. In conclusion, partial replacement of L-lysine·HCl with LCM from 0 to 1% had no negative impacts on the growth performance, but 100% replacement of L-lysine·HCl with LCM decreased the growth performance of weaning pigs. Therefore, LCM could be included in the diets for weaning pigs up to 2.8% and 1.76% for phase 1 and phase 2, respectively, as a substitute for L-lysine·HCl without detrimental effects on the performance of weaning pigs.

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

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

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.

Nutrient digestibility and fecal characteristics, microbiota, and metabolites in dogs fed human-grade foods.

Human-grade (HG) pet foods are commercially available, but they have not been well studied. Our objective was to determine the apparent total tract digestibility (ATTD) of HG pet foods and evaluate their effects on fecal characteristics, microbiota, and metabolites, serum metabolites, and hematology of dogs. Twelve dogs (mean age = 5.5 ± 1.0; BW = 11.6 ± 1.6 kg) were used in a replicated 4 × 4 Latin square design (n = 12/treatment). The diets included 1) Chicken and Brown Rice Recipe (extruded; Blue Buffalo); 2) Roasted Meals Tender Chicken Recipe (fresh; Freshpet); 3) Beef and Russet Potato Recipe (HG beef; JustFoodForDogs); and 4) Chicken and White Rice Recipe (HG chicken; JustFoodForDogs). Each period consisted of 28 d, with a 6-d diet transition phase, 16 d of consuming 100% of the diet, a 5-d phase for fecal collection, and 1 d for blood collection. All data were analyzed using the Mixed Models procedure of SAS 9.4. Dogs fed the extruded diet required a higher (P < 0.05) daily food intake (dry matter basis, DMB) to maintain BW. The ATTD of dry matter (DM), organic matter (OM), energy, and acid-hydrolyzed fat (AHF) were greater (P < 0.05) in dogs fed the HG diets than those fed the fresh diet, and greater (P < 0.05) in dogs fed the fresh diet than those fed the extruded diet. Crude protein ATTD was lower (P < 0.05) for dogs fed the extruded diet than those fed all other diets. Dogs fed the extruded diet had greater (P < 0.05) fecal output (as-is; DMB) than dogs fed fresh (1.5-1.7 times greater) or HG foods (2.0-2.9 times greater). There were no differences in fecal pH, scores, and metabolites, but microbiota were affected by diet. Dogs fed HG beef had higher (P < 0.05) relative abundance of Bacteroidetes and lower (P < 0.05) relative abundance of Firmicutes than dogs fed the fresh or HG chicken diets. The Actinobacteria, Fusobacteria, Proteobacteria, and Spirochaetes phyla were unchanged (P > 0.05), but diet modified the relative abundance of nearly 20 bacterial genera. Similar to previous reports, these data demonstrate that the fecal microbiota of dogs fed HG or fresh diets is markedly different than those consuming extruded diets, likely due to ingredient, nutrient, and processing differences. Serum metabolites and hematology were not greatly affected by diet. In conclusion, the HG pet foods tested resulted in significantly reduced fecal output, were highly digestible, maintained fecal characteristics, serum chemistry, and hematology, and modified the fecal microbiota of dogs.

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.

End point determination of blending process for trimebutine tablets using principle component analysis (PCA) and partial least squares (PLS) regression.

This study showed near Infrared (NIR) and Raman spectroscopy with a multivariate calibration approach were very effective to determine blend uniformity end-point. A set of 36 trimebutine samples containing magnesium stearate, stearic acid, colloidal silicon oxide, talc as excipients (0.9%∼1.8%) was acquired from six positions during blending processing with U-type blender from 0 to 30 min. Principle component analysis (PCA) with NIR and Raman spectral data was used to confirm the end-point of blending. After 30 min, the scores of principle component (PC) 1 and principle component (PC) 2 for samples moved into one point, which clearly indicated the mixture of sample became homogenous. In addition, NIR and Raman spectroscopy has been applied to the quantitative analysis of 20 trimebutine samples containing 2∼40% in mixture granules, which divided into a calibration set of 15 samples and a prediction set of 5 samples for NIR spectral data. The standard error of calibration (SEC) and standard error of prediction (SEP) are 0.15% and 0.13%, respectively using NIR while SEC and SEP of 0.95% and 0.91% are obtained using Raman spectroscopy. The results showed the NIR and Raman spectroscopy with a multivariate calibration such as PCA and PLS provide the possibility of real time monitoring of homogeneity and content uniformity during blending process.