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Background Dietary interventions are thought to modify gut microbial communities in healthy individuals. In dogs with chronic enteropathies, resolution of dysbiosis, along with remission of clinical signs, is expected with treatment. Hypothesis/Objective To evaluate changes in the fecal microbiota in dogs with food‐responsive chronic enteropathy (FRE) and in healthy control (HC) dogs before and after an elimination dietary trial with an animal protein‐free diet (APFD). Animals Dogs with FRE (n = 10) and HC (n = 14). Methods Dogs were fed the APFD for 60 days. Fecal microbiota was analyzed by Illumina 16S rRNA sequencing and quantitative polymerase chain reaction (PCR). Results A significantly lower bacterial alpha‐diversity was observed in dogs with FRE compared with HC dogs at baseline, and compared with FRE dogs after the trial. Distinct microbial communities were observed in dogs with FRE at baseline compared with HC dogs at baseline and compared with dogs with FRE after the trial. Microbial communities still were different in FRE dogs after the trial compared with HC dogs at baseline. In HC dogs, the fecal microbiota did not show a significant modification after administration of the APFD. Conclusion and Clinical Importance Our results suggest that, in FRE dogs, treatment with the APFD led to a partial recovery of the fecal microbiota by significantly increasing microbiota richness, which was significantly closer to a healthy microbiota after the treatment. In contrast, no changes were detected in the fecal microbiota of HC dogs fed the same APFD.

Background Dietary protein and phosphorus (P) restriction is the mainstay for nutritional management of chronic kidney disease (CKD). However, adequate restriction levels for cats with early CKD remain unclear. Objectives To investigate responses in cats with early CKD to varying dietary protein, P, and calcium (Ca) : P ratio. Animals Nineteen research colony cats with International Renal Interest Society stages 1‐2 CKD. Methods In an opportunistic longitudinal case study, cats were fed a low protein (59 g/Mcal), low P (0.84 g/Mcal) dry diet (LP‐LP; Ca : P = 1.9) for 18 months and later transitioned onto a moderate protein (76‐98 g/Mcal), moderate P (1.4‐1.6 g/Mcal) dry‐wet diet regimen (MP‐MP; Ca : P = 1.4‐1.6) for 22 months. Fold‐changes in serum creatinine, total Ca (tCa) and P (primary outcomes) and fibroblast growth factor 23 (FGF23) were assessed by linear‐mixed models. Results While feeding LP‐LP, mean serum creatinine decreased (0.87‐fold, 95% confidence interval [CI] 0.81, 0.93, P < .001) to within reference range after 6 months, while increases in total Ca (tCa; 1.16‐fold, 95% CI 1.11, 1.22, P < .001) and FGF23 (2.72‐fold, 95% CI 1.72, 4.31, P < .001), but not in P (1.03‐fold, 95% CI 0.945, 1.124, P = .94), were observed after 17 months. On MP‐MP, mean creatinine, tCa and P remained within reference ranges and did not significantly change (P = .11, P = .98, and P = 1, respectively), while FGF23 significantly decreased (0.58‐fold, 95% CI 0.36, 0.95, P = .02) after 22 months. Conclusions and Clinical Importance Cats with early CKD developed hypercalcemia after long‐term feeding of a highly P‐restricted diet. Increasing dietary P and reducing Ca : P ratio maintained renal markers, while improving Ca‐P balance. Cats with early CKD could benefit from moderately protein‐ and P‐restricted diets.

Reducing the levels of dietary protein is an effective nutritional approach in lowering feed cost and nitrogen emissions in ruminants. The purpose of this study was to evaluate the effects of dietary Lys/Met ratio in a low protein diet (10%, dry matter basis) on the growth performance and hepatic function (antioxidant capacity, immune status, and glycolytic activity) in Tibetan lambs. Ninety two-month-old rams with an average weight of 15.37 ± 0.92 kg were randomly assigned to LP-L (dietary Lys/Met = 1:1), LP-M (dietary Lys/Met = 2:1) and LP-H (dietary Lys/Met = 3:1) treatments. The trial was conducted over 100 d, including 10 d of adaption to the diets. Hepatic phenotypes, antioxidant capacity, immune status, glycolytic activity and gene expression profiling was detected after the conclusion of the feeding trials. The results showed that the body weight was higher in the LP-L group when compared to those on the LP-M group ( P  < 0.05). In addition, the activities of the catalase (CAT) and glutathione peroxidase (GSH-Px) in the LP-L group were significantly increased compared with the LP-M group ( P  < 0.05), while the malondialdehyde (MDA) levels in LP-H group were significantly decreased ( P  < 0.05). Compared with LP-H group, both hepatic glycogen ( P  < 0.01) and lactate dehydrogenase (LDH) ( P  < 0.05) were significantly elevated in LP-L group. For the LP-L group, the hepatocytes were arranged radially with the central vein in the center, and hepatic plates exhibited tight arrangement. Transcriptome analysis identified 29, 179, and 129 differentially expressed genes (DEGs) between the LP-M vs. LP-L, LP-H vs. LP-M, and LP-H vs. LP-L groups, respectively (Q-values < 0.05 and |log2Fold Change| > 1). Gene Ontology (GO) and correlation analyses showed that in the LP-L group, core genes ( C1QA and JUNB ) enriched in oxidoreductase activity were positively correlated with antioxidant indicators, while the MYO9A core gene enriched in the immune response was positively associated with immune indicators, and core genes enriched in molecular function ( PDK3 and PDP2 ) were positively correlated with glycolysis indicators. In summary, low-protein diet with a low Lys/Met ratio (1:1) could reduce the hepatic oxidative stress and improve the glycolytic activity by regulating the expression of related genes of Tibetan sheep.

For this study, threonine (Thr) deficiency was hypothesised to exacerbate the intestinal damage induced by feed withdrawal with coccidial infection because of its high obligatory requirement by the gut; two dietary Thr treatments (0·49 and 0·90 %) were applied to chicks from 0 to 21 d of age. At 13 d of age, feed was withdrawn for 24 h from one-half of birds of each dietary treatment with subsequent gavage of a 25× dose of coccidial vaccine. Overall, there were four treatments with eight replicate cages per treatment. Under combined challenge, birds fed the Thr-deficient diet had 38 % lower 13–21-d body weight gain (P≤0·05) compared with birds fed the Thr-control diet. At 21 d, the challenged group fed low Thr had higher number of oocysts (+40 %, P=0·03) and lower crypt depth (−31 %, P<0·01). In addition, birds fed the low-Thr diet had higher gut permeability as measured after 2 h of administration of fluorescein isothiocyanate-dextran (3–5 kDa, P<0·01), which may be attributed to decreased IgA production (P=0·03) in the ileum. In caecal tonsils, the challenged group fed low Thr had lower CD3:Bu-1 ratio (P≤0·05), along with a tendency for lower CCR9 mRNA expression in birds fed the low-Thr diet (P=0·10). In addition, Thr deficiency tended to increase IL-10 mRNA expression regardless of infection (P=0·06), but did not change interferon-γ mRNA expression upon coccidial infection (P>0·05). Overall, Thr deficiency worsened the detrimental effects of combined feed withdrawal and coccidial infection on growth performance and oocyst shedding by impairing intestinal morphology, barrier function, lymphocyte profiles and their cytokine expressions.

Skeletal muscle is a major site for the oxidation of fatty acids (FA) in mammals, including humans. Using a swine model, we tested the hypothesis that dietary protein intake regulates the expression of key genes for lipid metabolism in skeletal muscle. A total of ninety-six barrows (forty-eight pure-bred Bama mini-pigs (fatty genotype) and forty-eight Landrace pigs (lean genotype)) were fed from 5 weeks of age to market weight. Pigs of fatty or lean genotype were randomly assigned to one of two dietary treatments (low- or adequate-protein diet), with twenty-four individually fed pigs per treatment. Our data showed that dietary protein levels affected the expression of genes involved in the anabolism and catabolism of lipids in the longissimus dorsi and biceps femoris muscles in a genotype-dependent manner. Specifically, Bama mini-pigs had more intramuscular fat, SFA and MUFA, as well as elevated mRNA expression levels of lipogenic genes, compared with Landrace pigs. In contrast, Bama mini-pigs had lower mRNA expression levels of lipolytic genes than Landrace pigs fed an adequate-protein diet in the growing phase. These data are consistent with higher white-fat deposition in Bama mini-pigs than in Landrace pigs. In conclusion, adequate provision of dietary protein (amino acids) plays an important role in regulating the expression of key lipogenic genes, and the growth of white adipose tissue, in a genotype- and tissue-specific manner. These findings have important implications for developing novel dietary strategies in pig production.

Abstract The supplementation of dietary limiting amino acids (AA) with crystalline AA makes the use of low-protein diets an option in poultry production. The differing absorption rates of crystalline and protein-bound AA may lead to temporally imbalanced AA in the postabsorptive period. In this study, two experiments were conducted to evaluate the effect of encapsulated L-lysine-HCl (L-Lys-HCl) and DL-methionine (DL-Met) on the laying performance of hens. In exp. 1, a total of 135 forty-seven-wk-old Hy-Line Brown hens were subjected to three dietary treatments for 8 wk: basal diet supplemented with 0.14% L-Lys-HCl and 0.17% DL-Met to satisfy the NRC (1994) total Lys and Met recommendation (control) and basal diet supplemented with encapsulated L-Lys-HCl and DL-Met at the levels of 60% (60CLM, 0.084% L-Lys-HCl and 0.102% DL-Met) or 80% of control (80CLM, 0.112% L-Lys-HCl and 0.136% DL-Met), respectively. In exp. 2, 24 fifty-five-wk-old Hy-Line Brown hens were individually reared in cages and subjected to the same treatments as in exp. 1. The plasma concentrations of free AA and nitrogen metabolites were measured 2, 4, and 6 h after fed. The results showed that dietary AA treatment had no significant influence on body weight (BW), feed intake, laying rate, egg weight, egg mass, or feed efficiency. The expression levels of AA transporters CAT-1, y+LAT1, b0,+AT, B0AT, rBAT, EAAT3, and PepT1 in the duodenum, jejunum, and ileum were not influenced (P > 0.05) by dietary treatment. There was an interaction of dietary AA treatment and time (P < 0.05) and the 80CLM hens exhibited higher concentrations of Lys (P < 0.05) than the controls at 2-h time point. In contrast, plasma Met concentration was not influenced (P > 0.05), while Cys was reduced in the 60CLM hens at every time point. The 80CLM hens had higher taurine concentrations than those receiving the control diet at every postprandial time point. In conclusion, these findings demonstrate that by using encapsulated form, the supplemental levels of synthetic L-Lys-HCl and DL-Met can be effectively reduced by approximately 20% with no negative effect on laying performance. The result suggests that encapsulated Lys and Met may ameliorate the postabsorptive AA balance and contribute to the reduced dietary AA supplemental levels.

The objective of the present study was to evaluate the effects of increasing doses of protease on broilers from 1 to 42 days of age. A total of 1290 Ross AP broilers were used, distributed among five treatments: positive control diet, negative control diet (NC), NC + 50 ppm of protease, NC + 100 ppm of protease, and NC + 200 ppm of protease. Each treatment contained six replicates of 43 animals each. The inclusion of proteases in the diet had effects (P < 0.05) on body weight, feed intake, weight gain, and feed conversion in the 12 to 21 day period; body weight, weight gain, and feed intake in the 29 to 42 day period; nutrient digestibility (energy metabolizability coefficient and crude protein at 28 days); and intestinal parameters (crypt and muscle width of jejunum and ileum at 28 days and villus length, crypt length, and jejunum thickness muscle layer at 42 days). These results indicate that the inclusion of protease in broiler feed can improve production parameters when the amount of crude protein in the diet is reduced.

Abstract A total of 240 newly weaned pigs (5.25 ± 0.15 kg BW) were used to determine the dietary omega-6-to-omega-3 (ω-6:ω-3) fatty acid ratio that optimized growth performance and immune responses when fed corn and soybean meal (SBM)-based diets with low protein quality. Pigs were randomly assigned to 1 of 5 dietary treatments (n = 6 pens per treatment; day 0 of study): [1] positive control (High; included animal proteins and 5% corn oil), [2] negative control (Low0; corn- and SBM-based and 5% corn oil), or 1 of 3 Low diets with increasing supplementation of fish oil to replace corn oil: [3] 1.25% (Low1.25), [4] 2.5% (Low2.5), [5] 5% (Low5) to achieve 5:1, 3:1, and 1:1 ω-6:ω-3 ratios, respectively. Pigs were fed dietary treatments in 2 phases for 7 and 14 d, respectively, followed by a common phase III diet for 21 d. On day 6 and 20, 12 pigs per treatment were immune sensitized with 0.5 mg ovalbumin (OVA) and 0.5 mg Quil A adjuvant in 1 mL saline. The dermal hypersensitivity response (DHR) was evaluated on day 40 in these same pigs, using intradermal injection of OVA; changes in skin-fold thickness were measured. On day 21, 4 pigs per pen were immune challenged with LPS (30 µg Escherichia coli LPS per kg BW) or saline (n = 12); rectal temperature was monitored over 3 h. During phase I only, ADG, ADFI, and G:F were greater for pigs fed the High diet vs. those fed the Low diet (P < 0.05), and increased with increasing fish oil supplementation up to 2.5% (Low2.5), but decreased for pigs fed the Low5 diet (quadratic; P < 0.05, P = 0.086, and P < 0.05 for ADG, ADFI, and G:F, respectively). On day 21, LPS increased rectal temperature (vs. saline at 1-, 2-, and 3-h post-challenge; P < 0.001); fish oil supplementation reduced rectal temperature 2-h post-challenge in the Low-fed pigs (linear; P < 0.05). On day 22, serum haptoglobin was greatest for pigs fed Low0 and decreased with increasing fish oil supplementation (linear; P < 0.05). Immunization with OVA induced a serum anti-OVA IgG response, which was reduced on day 34 among pigs fed Low diets with increasing fish oil supplementation (linear; P = 0.050). On day 40, and 6 h after intradermal injection of OVA, the DHR was least for pigs fed the Low2.5 diet (P < 0.05). Inclusion of 2.5% fish oil (3:1, ω-6:ω-3) optimized growth performance during the early nursery phase when pigs were most sensitive to diets with low protein quality; the ideal ω-6-to-ω-3 fatty acid ratio may differ when using immune responses as the major outcome.

Pigs fed protein-bound AA appear to have a higher abundance of AA transporters for their absorption in the jejunum compared with the duodenum. However, there is limited data about the effect of dietary free AA, readily available in the duodenum, on the duodenal abundance of AA transporters and its impact on pig performance. Forty-eight pigs (24.3 kg initial BW) distributed in 4 treatments were used to evaluate the effect of the CP level and form (free vs. protein bound) in which AA are added to diets on the expression of AA transporters in the 3 small intestine segments, serum concentration of AA, and performance. Dietary treatments based on wheat and soybean meal (SBM) were 1) low-CP (14%) diet supplemented with L-Lys, L-Thr, DL-Met, L-Leu, L-Ile, L-Val, L-His, L-Trp, and L-Phe (LPAA); 2) as in the LPAA but with added L-Gly as a N source (LPAA+N); 3) intermediate CP content (16%) supplemented with L-Lys HCl, L-Thr, and DL-Met (MPAA); and 4) high-CP (22%) diet (HP) without free AA. At the end of the experiment, 8 pigs from LPAA and HP were sacrificed to collect intestinal mucosa and blood samples and to dissect the carcasses. There were no differences in ADG, ADFI, G:F, and weights of carcass components and some visceral organs between treatments. Weights of the large intestine and kidney were higher in HP pigs (P < 0.01). Expression of b(0,+) in the duodenum was higher in pigs fed the LPAA compared with the HP diet (P= 0.036) but there was no difference in the jejunum and ileum. In the ileum, y+ L expression tended to be higher in pigs fed the LPAA diet (P = 0.098). Expression of b(0,+) in LPAA pigs did not differ between the duodenum and the jejunum, but in HP pigs, the expression of all AA transporters was higher in the jejunum than in the duodenum or ileum (P < 0.05). The serum concentration of Arg, His, Ile, Leu, Phe, and Val was higher but serum Lys and Met were lower in pigs fed the HP diet (P < 0.05). These results indicate that LPAA can substitute up to 8 percentage units of protein in HP wheat-SBM diets without affecting pig performance; nonessential N does not seem to be limiting in very low-protein wheat-SBM diets for growing pigs. Also, the inclusion of free AA in the diet appears to affect their serum concentration and the expression of the AA transporter b0,+ in the duodenum of pigs.

This study evaluated the effect of feeding low protein (LP) diets for 7 or 14 d after weaning or a high protein (HP) diet for 14 d after weaning on postweaning diarrhea (PWD), indices of protein fermentation, and production in pigs infected or not infected per os with an enterotoxigenic strain of Escherichia coli. A total of 72 female pigs weaned at aged 21 d with initial BW of 5.9 ± 0.12 kg were used in a 3 x 2 factorial arrangement of treatments. The factors were 3 feeding regimens associated with different combinations of feeding duration and diet CP level: (i) HP diet (256 g of CP/kg) fed for 14 d after weaning, (ii) LP diet (175 g of CP/kg) fed for 7 d after weaning, and (iii) LP diet fed for 14 d after weaning; and infection or noninfection with an enterotoxigenic strain of E. coli (10⁷ cfu/mL, serotype O149:K91:K88) at 72, 96, and 120 h after weaning. The LP diets were fortified with crystalline Ile and Val to achieve an ideal AA pattern. A second-stage diet (213 g of CP/kg) was fed to pigs at the conclusion of each feeding regimen, and the study finished 4 wk after weaning. None of the diets contained antimicrobials. Feeding the LP diets decreased (P < 0.001) plasma urea nitrogen, fecal ammonia nitrogen concentrations, and the incidence of PWD, but increased (P = 0.001) fecal DM content compared with pigs fed HP in the 2-wk period after weaning. Infection increased shedding of β-hemolytic E. coli (P < 0.001), the incidence of PWD (P < 0.001), and fecal ammonia nitrogen concentrations (P < 0.01), but did not interact with feeding regimen, after weaning. Pigs challenged with E. coli grew more slowly (P < 0.001) and had decreased G:F (P < 0.01) compared with nonchallenged pigs in the 4-wk period after weaning. Feeding an LP diet for 7 or 14 d after weaning markedly reduced the incidence of PWD after infection with β-hemolytic E. coli. Infection was associated with decreased indices of protein fermentation in the distal gastrointestinal tract but did not compromise the growth of weaner pigs in the 4-wk period after weaning.