Implications of excreta uric acid concentrations in broilers offered reduced-crude protein diets and dietary glycine requirements for uric acid synthesis.

In a previous experiment, male Ross 308 broiler chickens were offered dietary treatments with 3 levels of crude protein (222, 193, 165 g/kg) and 3 feed grains (ground maize, ground wheat, whole wheat) from 7 to 35 d post-hatch. Maize-based diets supported superior growth performance in comparison to wheat-based diets. Uric acid concentrations in excreta were retrospectively determined and related to total nitrogen (N) excreta concentrations. Uric acid concentrations ranged from 28.5 to 69.4 mg/g and proportions of uric acid-N to total excreta-N ranged from 27.4% to 42.6% in broiler chickens offered the 3 × 3 factorial array of dietary treatments. Proportions of uric acid-N to total N in excreta in birds offered the 165 g/kg CP, maize-based diet were significantly lower by 10.6 percentage units (27.4% versus 38.0%; P = 0.00057) than their wheat-based counterparts. Total excreta analysed had been collected from 35 to 37 d post-hatch when feed intakes and excreta outputs were monitored. There were linear relationships between proportions of uric acid-N to total N in excreta in birds offered the three 165 g/kg CP diets with weight gain (r = -0.587; P = 0.010), feed intake (r = -0.526; P = 0.025) and feed conversion ratios (r = 0.635; P = 0.005). The possibility that increasing uric acid-N proportions in excreta is indicative of excessive ammonia accumulations compromising growth performance is discussed. The mean proportion of dietary glycine involved in uric acid excretion was 49.2% across all dietary treatments but ranged from 25.0% to 80.9%. Thus, the appropriate amount of dietary glycine is variable and largely dependent on the volume of uric acid synthesised and excreted.

Utilization of Methionine Sources for Growth and Met+Cys Deposition in Broilers.

Knowledge about the nutritional value of methionine sources is highly important for their appropriate application in terms of animal and economic performance. Therefore, a broiler feeding trial was conducted to determine the relative bioavailability value (RBV) of DL-2-hydroxy-4-methylthio butanoic acid (HMTBA) compared to DL-methionine (DLM). DLM diluted to 65% purity (DLM65) served as the internal standard, with a known RBV of 65%. A total of 1920 d-old male broilers were used in the three-phase feeding trial comprising 16 treatments including a basal, Met+Cys-deficient diet and 5 graded DLM, HMTBA, or DLM65 levels. Growth performance and carcass quality data were subjected to multi-exponential regression analysis. Increasing levels of any Met source significantly improved all performance parameters compared to the negative control (p < 0.05). Across all performance parameters, the RBV of HMTBA was 63% and that of DLM65 was 58%. All RBV estimates of HMTBA and DLM65 were significantly lower than 88% (p < 0.05). Cumulative efficiency of DLM for Met+Cys deposition in body protein was higher than that of HMBTA at any dose, confirming the determined RBV. Using DLM65 as an internal marker allowed for validation of the methodology.

Modulations of genes related to gut integrity, apoptosis, and immunity underlie the beneficial effects of Bacillus amyloliquefaciens CECT 5940 in broilers fed diets with different protein levels in a necrotic enteritis challenge model.

BACKGROUND: The ban of in-feed antimicrobial additives has negatively affected the poultry industry by causing necrotic enteritis (NE) to emerge in the flocks. Alternatives such as Bacillus probiotics have shown to be effective on eliminating the negative effects of this disease. Two experiments were conducted to investigate the effect of Bacillus amyloliquefaciens CECT 5940 (BA) in broiler chickens under NE challenge and/or fed diets with different protein levels. METHODS: In both experiments, 480 day-old mix-sexed Ross-308 broilers were arranged in a 2 × 2 factorial arrangement of treatments. In experiment 1, the factors were NE challenge (yes or no) and probiotic (yes or no). In experiment 2, the factors were dietary crude protein levels (standard or reduced) and probiotic (yes or no) and were used under NE challenge condition. Oral administration of Eimeria oocysts (day 9) followed by inoculation with Clostridium perfringens (day 14 and 15) was used to induce NE challenge. On day 16, two birds from each treatment were gavaged with fluorescein isothiocyanate-dextran (FITC-d) and blood samples were collected for gut integrity evaluation, and jejunal samples were collected for gene expression assay. RESULTS: In experiment 1, BA supplementation decreased caspase-3 (CASP3) (P < 0.001) and caspase-8 (CASP8) (P < 0.05) and increased occludin (OCLD) (P < 0.05) expression regardless of the challenge. Additionally, BA supplementation downregulated interfron-γ (IFN-γ) expression (P < 0.01) and upregulated immunoglobulin-G (IgG) (P < 0.01) and immunoglobulin-M (IgM) (P < 0.05) only in challenged birds. In experiment 2, the expression of genes encoding mucin-2 (MUC2) (P < 0.001), tight junction protein-1 (TJP1) (P < 0.05) and OCLD (P < 0.05) were upregulated by the addition of BA in the diet, regardless of the crude protein level. Further, BA supplementation downregulated INF-γ (P < 0.01) and upregulated immunoglobulin-A (IgA) (P < 0.05), IgM (P < 0.05) and IgG (P < 0.01) regardless of the crude protein level. CONCLUSION: These findings suggest that supplementation of BA in broiler diets can improve gut health by modulation of genes related to the mucosal barrier, tight junction, and immunity in broilers challenged by unfavourable conditions such as NE challenge.

Modelling of sulphur amino acid requirements and nitrogen endogenous losses in kittens.

The objective of this study was to estimate the sulphur amino acid (methionine + cystine) requirements and nitrogen endogenous losses in kittens aged 150 to 240 d. Thirty-six cats were distributed in six treatments (six cats per treatment) consisting of different concentrations of methionine + cystine (M + C): T1, 6.5 g/kg; T2, 8.8 g/kg; T3, 11.3 g/kg; T4, 13.6 g/kg; T5, 16.0 g/kg; and control, 6.5 g/kg. Diets were formulated by serial dilution of T5 (a diet relatively deficient in M + C but containing high protein concentrations) with a minimal nitrogen diet (MND). Thus, crude protein and amino acid concentrations in diets T1-T5 decreased by the same factor. The control diet was the T1 diet supplemented with adequate concentrations of M + C (6.5 g/kg; 8.8 g/kg; 11.3 g/kg; 13.6 g/kg and 16.0 g/kg). All diets were based on ingredients commonly used in extruded cat diets. Digestibility assays were performed for the determination of nitrogen balance. Nitrogen intake (NI) and nitrogen excretion (NEX) results data were fitted with an exponential equation to estimate nitrogen maintenance requirement (NMR), theoretical maximum for daily nitrogen retention (NRmaxT), and protein quality (b). M + C requirements were calculated from the limiting amino acid intake (LAAI) equation assuming a nitrogen retention of 45 to 65% NRmaxT. The NMR of kittens aged 150, 195, and 240 d was estimated at 595, 559, and 455 mg/kg body weight (BW)0.67 per day, respectively, and M + C requirements were estimated at 517, 664, and 301 mg/kg BW0.67 per day, respectively.

Synthetic and Crystalline Amino Acids: Alternatives to Soybean Meal in Chicken-Meat Production.

: This review explores the premise that non-bound (synthetic and crystalline) amino acids are alternatives to soybean meal, the dominant source of protein, in diets for broiler chickens. Non-bound essential and non-essential amino acids can partially replace soybean meal so that requirements are still met but dietary crude protein levels are reduced. This review considers the production of non-bound amino acids, soybeans, and soybean meal and discusses the concept of reduced-crude protein diets. There is a focus on specific amino acids, including glycine, serine, threonine, and branched-chain amino acids, because they may be pivotal to the successful development of reduced-crude protein diets. Presently, moderate dietary crude protein reductions of approximately 30 g/kg are feasible, but more radical reductions compromise broiler performance. In theory, an 'ideal' amino acid profile would prevent this, but this is not necessarily the case in practice. The dependence of the chicken-meat industry on soybean meal will be halved if crude protein reductions in the order of 50 g/kg are attained without compromising the growth performance of broiler chickens. In this event, synthetic and crystalline, or non-bound, amino acids will become viable alternatives to soybean meal in chicken-meat production.

Nitrogen maintenance requirements and potential for nitrogen retention of pullets in growth phase.

Experiments were conducted to estimate daily N maintenance requirements (NMRs) and the genetic potential for daily N retention (NRmax T) of pullets in growth phase. Three nitrogen balance trials were conducted, and a total of 48 Hy-line W-36 pullets were used in each trial in age periods (starter: 14-28, grower: 56-70 and developer: 98-112 days). The treatments consisted of six graded levels of nitrogen in the diets (L1 = 8, L2 = 16, L3 = 24, L4 = 32, L5 = 40 and L6 = 48 g N/kg of feed), formulated using the dilution technique. The regression analyses between nitrogen intake and nitrogen excretion were performed to fit the exponential function and to determine the NMR. The daily NMRs that were estimated at 294, 331 and 355 mg/BWkg 0.67 for the initial, grower and developer periods, respectively, were applied for further calculation of NRmax T as the threshold value of the function between N intake and daily N balance. The NRmax T was estimated by a statistical procedure following several iteration steps by the Levenberg-Marquardt algorithm until the sum of the squares of the residual was minimized. The NRmax T was estimated at 3,200, 2,633 and 1,826 mg/BWkg 0.67 for starter, grower and developer periods respectively. The determined model parameters were the precondition for modelling of the amino acid requirement based on an exponential N-utilization model and depended on performance and dietary amino acid efficiency. This procedure will be further developed and applied in the subsequent study.

Bacillus amyloliquefaciens CECT 5940 alone or in combination with antibiotic growth promoters improves performance in broilers under enteric pathogen challenge.

A study was conducted to investigate the effects of Bacillus amyloliquefaciens CECT 5940 as a direct-fed microbial (DFM) alone or in association with bacitracin methylene disalicylate (BMD) in broilers under enteric pathogen challenge. A total of 1,530-day-old male Cobb500 chicks were randomly assigned to 5 treatments, with 9 replicate pens with 34 birds each. Treatments included positive control (PC, basal diet without additives or challenge); negative control (NC, basal diet without additive and challenged birds); NC + 0.05 g/kg BMD; NC + 1 g/kg DFM (106 CFU B. amyloliquefaciens CECT 5940/g of feed); and NC + 0.05 g/kg BMD + 1 g/kg DFM. The challenge consisted of oral gavage with Eimeria maxima and Clostridium perfringens inoculum. Body weight gain (BWG), feed intake (FI), and feed conversion ratio (FCR) were evaluated on days 21, 35, and 42. Ileal and cecal content were collected on days 21 and 28 for C. perfringens enumeration by real-time PCR assay and the intestinal health was evaluated by scores. Uniformity (UN), carcass (CY), and breast meat yields (BMY) were evaluated on day 42. After 14 and 21 d post-inoculation, birds in the challenged groups had significant lower FI and BWG compared to the PC group (P < 0.05). However, the groups receiving DFM, BMD, or its combination presented better FCR, CY, BMY, UN, and lower incidence of footpad lesion and litter quality visual scores, compared to the NC group without feed additives (P < 0.05). Mortality was not affected by treatments (P > 0.05). Broilers fed DFM, BMD, or its combination presented lower C. perfringens in ileal content at 21 and 28 d compared to NC group without additives (P < 0.05) and also maintained gut health by keeping the frequency of ballooning, abnormal content, and swollen mucosa comparable to the PC group (P > 0.05). The study indicates that Bacillus amyloliquefaciens CECT 5940 is effective as BMD to provide similar performance and gut health in challenged broilers.

Effects of leucine supplementation on muscle protein synthesis and degradation pathways in broilers at constant dietary concentrations of isoleucine and valine.

The present study investigated the hypothesis that dietary concentrations of leucine (Leu) in excess of the breeder´s recommendations activates protein synthesis and decreases protein degradation in muscle of broilers. Day-old male Ross 308 broilers (n = 450) were phase-fed corn-soybean meal-based diets during starter (d 1-10), grower (d 11-22), and finisher (d 23-34) period. The basal diets fed to the control group (L0) met the broilers' requirements for nutrients and amino acids, and contained Leu, Leu:isoleucine (Ile) and Leu:valine (Val) ratios, close to those recommended by the breeder (Leu:Ile: 100:54, 100:52, 100:51; Leu:Val 100:64, 100:61, 100:58; in starter, grower and finisher diet, resp.). Basal diets were supplemented with Leu to exceed the breeder's recommendations by 35% (group L35) and 60% (group L60). Growth performance during 34 d, and carcass weights, and breast and thigh muscle weights on d 34 were similar among groups. Hepatic and muscle mRNA levels of genes involved in the somatotropic axis [growth hormone receptor, insulin-like growth factor (IGF)-1, IGF binding protein 2, IGF receptor] on d 34 were not influenced by Leu. In the breast muscle, relative mRNA abundances of genes involved in the mammalian target of rapamycin (mTOR) pathway of protein synthesis (mTOR, ribosomal p70 S6 kinase) and the ubiquitin-proteasome pathway of protein degradation (F-box only protein 32, Forkhead box protein O1, Muscle RING-finger protein-1) on d 34 were largely similar among groups. Likewise, relative phosphorylation and thus activation of mTOR and ribosomal protein S6 involved in the mTOR pathway, and of eukaryotic translation initiation factor 2A (eIF2a) involved in the general control nonderepressible 2 (GCN2)/eIF2a pathway of protein synthesis inhibition, were not influenced. These data indicate that dietary Leu concentrations exceeding the broiler´s requirements up to 60% neither influence protein synthesis nor degradation pathways nor muscle growth in growing broilers.

Bacillus subtilis DSM 32315 Supplementation Attenuates the Effects of Clostridium perfringens Challenge on the Growth Performance and Intestinal Microbiota of Broiler Chickens.

The objective of this study was to evaluate the effects of the dietary supplementation of Bacillus subtilis DSM 32315 (probiotic) on the performance and intestinal microbiota of broiler chickens infected with Clostridium perfringens (CP). One-day-old broiler chickens were assigned to 3 treatments with 8 replicate pens (50 birds/pen). The treatments were: non-infected control; infected control; and infected supplemented with probiotic (1 × 106 CFU/g of feed). On day of hatch, all birds were sprayed with a coccidia vaccine based on the manufacturer recommended dosage. On d 18⁻20 the infected birds were inoculated with CP via feed. Necrotic enteritis (NE) lesion score was performed on d 21. Digestive tract of 2 birds/pen was collected on d 21 to analyze the ileal and cecal microbiota by 16S rRNA sequencing. Performance was evaluated on d 28 and 42. On d 21, probiotic supplementation reduced (p < 0.001) the severity of NE related lesion versus infected control birds. On d 28, feed efficiency was improved (p < 0.001) in birds supplemented with probiotic versus infected control birds. On d 42, body weight gain (BW gain) and feed conversion ratio (FCR) were improved (p < 0.001) in probiotic supplemented birds versus infected control birds. The diversity, composition and predictive function of the intestinal microbial digesta changed with the infection but the supplementation of probiotic reduced these variations. Therefore, dietary supplementation of Bacillus subtilis DSM 32315 was beneficial in attenuating the negative effects of CP challenge on the performance and intestinal microbiota of broilers chickens.