Effects of dietary digestible amino acids and a novel exogenous sfericase protease on growth performance of broilers.

Two experiments (Exp.) were conducted to evaluate the effects of a novel exogenous sfericase protease on growth performance and ileal digestibility of broiler chickens until day 35 of age. In Exp. 1, 1350 one-day-old male chicks (Cobb 500) were allocated in 54 floor pens and fed one of the three dietary treatments, with 18 replicates of 25 birds each in a completely randomized design. Diets consisted of positive control [PC; commercially relevant ME and balanced amino acids (AA)]; negative control (NC; with reduction of 6% dig. Lys and proportional reductions for adjacent AA compared to the PC), and NC supplemented with sfericase protease [30,000 New Feed Protease units (NFP)/kg]. On day 35, ileal digesta was collected to determine apparent ileal digestibility of dry matter and nitrogen (N). In Exp. 2, 1620 one-day-old male chicks (Cobb 500) were allocated in 54 floor pens having three treatments and 18 replicates of 30 birds each in a completely randomized design from day 1-35. Broilers were fed a control basal diet (Control); Control supplemented with sfericase at 30,000 NFP/kg and at 60,000 NFP/kg. In Exp. 1, from day 1-35, body weight gain (BW gain) and feed conversion ratio (FCR) of broilers improved 3.4 and 2.5% when diets were supplemented with sfericase, respectively, whereas the digestibility of N increased by 2.7% compared to the NC. In Exp. 2, diets with usual protein and AA levels and supplemented with 30,000 NFP/kg had 2.3 and 1.75% improved BW gain and FCR from day 1-35, respectively. When diets were supplemented with 60,000 NFP/kg, BW gain and FCR were enhanced by 3.9 and 3.2%, respectively compared to the Control. In conclusion, these results demonstrate that the novel sfericase protease could be successfully used in corn-soy diets with protein and AA reductions or in feed formulations with usual digestible AA levels to enhance growth performance of broilers.

A microbial muramidase improves growth performance and reduces inflammatory cell infiltration in the intestine of broilers chickens under Eimeria and Clostridium perfringens challenge.

The objective of the present studies was to evaluate muramidase (MUR) supplementation in broilers under Eimeria and/or Clostridium perfringens challenge. For this, 2 experiments were conducted. Experiment 1. A total of 256 one-day old male Cobb 500 chicks were placed in battery cages in a completely randomized design, with 5 treatment groups, 7 replicate cages per treatment and 8 birds per cage. The treatments were: nonchallenged control (NC), challenged control (CC), CC + MUR at 25,000 or 35,000 LSU(F)/kg, and CC + Enramycin at 10 ppm (positive control-PC). Challenge consisted of 15× the recommended dose of coccidiosis vaccine at placement, and Clostridium perfringens (108 CFU/bird) inoculation at 10, 11, and 12 d. Macro and microscopic evaluation, immunohistochemistry, and gene expression were evaluated at 7, 14, 21, and 28 d of age. Experiment 2. A total of 1,120 one-day old male Cobb 500 chicks were placed in floor pens with fresh litter in a completely randomized design, with 4 treatment groups, 8 replicate pens per treatment, and 35 birds per pen. The treatments were: Control, supplementation of MUR at 25,000 or 45,000 LSU(F)/kg, and a positive control (basal diet plus Enramycin). At 10, 11, and 12 d of the experiment all the birds were inoculated by oral gavage with a fresh broth culture of a field isolate Clostridium perfringens (0.5 mL containing 106 CFU/bird). It was observed that in Experiment 1 MUR supplementation reduced the infiltration of macrophages and CD8+ lymphocytes in the liver and ileum of infected birds, downregulated IL-8 and upregulated IL-10 expression. In Experiment 2, MUR linearly improved the growth performance of the birds, increased breast meat yield, and improved absorption capacity. MUR supplementation elicited an anti-inflammatory response in birds undergoing a NE challenge model that may explain the improved growth performance of supplemented birds.

Effect of microbial muramidase supplementation in diets formulated with different fiber profiles for broiler chickens raised under various coccidiosis management programs.

The objective of the present study was to determine the effects of muramidase (MUR) supplemented to diets formulated with different fiber sources (inert or fermentable) on the growth performance and intestinal parameters of broiler chickens raised under different coccidiosis management programs. A total of 2,208 male Ross 308 broilers were housed in 96 floor pens and distributed into a 2 × 3 × 2 factorial arrangement in a completely randomized block design with 2 sources of fiber (inert or fermentable fiber), 3 coccidiosis management programs (none, vaccine, or Salinomycin), and with or without supplementation of MUR at 35,000 LSU(F)/kg of diet. Body weight gain (BWG), feed intake (FI), and feed conversion ratio (FCR) were calculated for each feeding phase (d 0-14, d 14-28, d 28-36) and from d 0 to 36. On d 17 and d 31, samples were taken to analyze several parameters. The experimental data were analyzed with 3-way ANOVA considering the main effect of fiber source, coccidiosis program, inclusion of MUR, and their interactions using JMP 16.2. 16S rDNA sequencing of the ileal and cecal content was carried out to analyze the diversity, composition, and predictive function of the microbiota. From d 0 to 36, BWG increased (P = 0.05) by 2.5% in birds supplemented with Salinomycin (P = 0.04), and by 2.2% with MUR supplementation (P = 0.02). Salinomycin and MUR improved FCR (P < 0.0001) when compared to nonsupplemented birds. The supplementation of MUR, regardless of the coccidiosis management program, reduced the intestinal viscosity (P = 0.03). On d 31, the highest blood concentration of carotenoids was observed in chickens fed diets supplemented with Salinomycin. MUR led to significant changes in the diversity, composition, and predictive function of the ileal microbiota, mainly on d 31. The results observed herein further explain the positive effects of MUR on the growth performance of broiler chickens.

The effect of amylase supplementation on individual variation, growth performance, and starch digestibility in broiler chickens.

The objective of this study was to evaluate the variance of starch digestibility in broilers individually fed diets without or with supplemental exogenous amylase. A total of 120 d-of-hatch male chicks were individually reared from 5 to 42 d in metallic cages and fed maize-based basal diets or diets containing 80 kilo-novo-α-amylase units/kg (60 birds or replicates per treatment). Beginning on d 7, feed intake, body weight gain, and feed conversion ratio were recorded; partial excreta collection was conducted every Monday, Wednesday, and Friday until 42 d, when all birds were sacrificed for individual collection of duodenal and ileal digesta. Lower feed intake (4,675 vs. 4,815 g) and feed conversion ratio (1.470 vs. 1.508) were observed in amylase-fed broilers during the overall period (7-43 d; P < 0.01), whereas body weight gain was not affected. Amylase supplementation improved total tract starch (TTS) digestibility (P < 0.05) on each day of excreta collection (except for d 28, where no difference was found), averaging 0.982 vs. 0.973 compared to basal-fed broilers from d 7 to 42. Both apparent ileal starch (AIS) digestibility and apparent metabolizable energy (AMEN) were increased (P <0.05) from 0.968 to 0.976 and from 3,119 to 3,198 kcal/kg, respectively, with enzyme supplementation. Activity of amylase in the duodenum was higher (18.6 vs. 50.1 IU/g of digesta) in supplemented birds. Amylase supplementation led to a reduced coefficient of variation for both TTS (averaged 2.41 vs. 0.92% from 7 to 42 d) and AIS digestibilities (1.96 vs. 1.03%), as well as AMEN (0.49 vs. 0.35%), when compared to the nonsupplemented group, indicating lower individual heterogenity. An age effect was detected for TTS digestibility, as both groups saw an increase during the first weeks (slightly more pronounced in the supplemented group); older birds (d 30 onwards) presented a lower TTS digestibility compared to ages between 7 and 25 d. In conclusion, amylase supplementation in maize diets for broilers can attenuate individual bird variation for starch and energy utilization by increasing amylase activity and enhancing starch digestibility.

Efficacy and safety profile of a subtilisin protease produced by fermentation in Bacillus licheniformis to be used as a feed additive.

The efficacy and safety of a novel Subtilisin protease from a Bacillus sp. produced in Bacillus licheniformis was investigated in broiler chickens, and a range of toxicological tests, respectively. The B. licheniformis production strain culture supernatant was not found cytotoxic in a Vero cell assay. Subtilisin was non-mutagenic and non-clastogenic in in-vitro tests, and did not exhibit irritating potential to the eye or skin in ex-vivo/in-vitro models. Oral administration of Subtilisin to rats did not cause any adverse effects in a 13-week sub-chronic toxicity study. In addition, a 35-day dose response broiler performance trial conducted with Subtilisin (30,000 and 60,000 NFP/kg diet), showed a significant linear improvement in both body weight gain and feed conversion ratio up to 35 days of protease supplementation. In conclusion, there are no safety concerns using this novel Subtilisin as a feed additive, and the protease is efficient in improving broiler growth performance, making it a good candidate for use as a feed additive.

Quantifying the effect of coccidiosis on broiler performance and infection outcomes in the presence and absence of control methods.

A systematic review and meta-analysis was conducted to investigate the role of Eimeria species, dose and inoculation time, on performance and infection outcomes of different broiler strains infected for different study durations. The meta-analysis addressed E. acervulina, E. maxima, E. tenella, and mixed species infections, and involved data from 72 peer-reviewed articles, corresponding to 521 treatments performed on 20,756 broilers. A secondary objective was to investigate the effects of synthetic anticoccidials, ionophores, and vaccination against Eimeria on the above outcomes. Performance during infection was scaled (%) to that of the uninfected birds. Infection reduced scaled ADFI and ADG (P < 0.001) and increased feed conversion ratio (FCR; P < 0.05); there was a significant interaction between dose and species on scaled ADFI and ADG, suggesting that different species affected these variables to different extents (P < 0.001). There was a tendency for an interaction between dose and broiler strain on scaled ADFI (P = 0.079), and a significant interaction between these variables on scaled ADG (P < 0.01). A tendency for an interaction between oocyst dose and Eimeria species (P = 0.067) on maximum number of oocysts excreted was observed. Lesion scores were significantly affected by dose, species, and their interaction (P < 0.05), the latter caused by an increase in the lesion scores during E. maxima and E. tenella infections. Control methods significantly affected scaled ADG and FCR (P < 0.05) and there was an interaction between dose and control methods on ADFI (P < 0.001). Synthetic anticoccidial use improved scaled ADG (P < 0.01), whereas ionophores improved FCR compared with untreated birds (P < 0.01). An interaction between dose and control method on scaled ADFI was caused by the higher ADFI of vaccinated compared to untreated birds, as dose increased. There was a significant effect of control methods on lesion scores (P < 0.01). All findings advance our understanding of the factors that influence the impact of coccidiosis and its controls in broilers.

A New Rapid and Quantitative Assay to Determine the Phytase Activity of Feed.

The fortification of animal feed with enzymes in order to optimize feed utilization has become a standard for the meat production industry. A method for measuring levels of active enzymes that can be carried out quickly would ensure that feed has been supplemented with the appropriate amount of enzyme. Phytase is the most widely used feed enzyme and is routinely quantified with an activity assay in a limited number of specialized laboratories. As an alternative, we report here the development of a rapid and easy method to perform a quantitative assay for the phytase from Citrobacter braakii. The method is suitable for use at local sites with a minimum lab setup and will reduce delays and potential interferences due to improper sample storage and shipment. The new assay is based on a lateral flow immunoassay that utilizes magnetic immune-chromatographic test (MICT) technology to quantify the phytase content of a feed extract. After extraction of the phytase from the feed, the sample is simply diluted and added to a reaction tube containing a specific anti-phytase antibody coupled to superparamagnetic particles. The mixture is then applied on an assay cassette, where the formed particle-antibody-phytase complexes are captured by immobilized antibodies on a nitro-cellulose strip housed in a cassette. The cassette is placed in the MICT reader that measures the magnetic signal of the captured particles. Using the calibration information stored in the cassette barcode, the signal is converted to a phytase concentration, given as phytase activity (FYT) per kilogram of feed. The accuracy and robustness of the assay compared to the ISO phytase activity assay were demonstrated through a large validation study including real feed samples from different compositions and origins. The MICT assay is the first quantitative assay for feed enzymes that is fast, reliable, and simple to use outside of a specialized reference laboratory and that is suitable for use in place of the current ISO assay.

Balanced nutrient density for broiler chickens using a range of digestible lysine-to-metabolizable energy ratios and nutrient density: Growth performance, nutrient utilisation and apparent metabolizable energy.

Currently, specific nutrient concentration, metabolizable energy (ME) and digestible amino acids are used as feed formulation criteria. A balanced nutrient density (BND) concept based on 2 criteria of nutrient density and balanced amino acids-to-ME ratio may offer more flexibility in optimisation of profit in formulation of diets compared with current formulation based on set values per unit of feed mass. A total of 672 one-d-old off-sex male Ross 308 broiler chickens were used across two 42-d performance trials in a 3 × 2 factorial arrangement of treatments with each diet replicated 8 times (14 birds per replicate). The experimental factors were 2 nutrient density levels (low [LD] and high [HD]) and 3 digestible lysine-to-ME ratios (DLYS:ME; low, medium, and high). Low density diets had ME of 2,876 and 3,023 kcal/kg for starter and finisher, respectively, while values for HD diets were 3,169 and 3,315 kcal/kg with proportionally higher non-nitrogenated nutrients. Separate digestibility and apparent metabolizable energy (AME) assays were conducted at d 21 and 42. Digestibility assays at d 7 were conducted on birds used for performance trials. Regardless of the diet density, birds fed low DLYS:ME had a lower (P < 0.01) feed intake (d 0 to 42) than medium and high DLYS:ME. Without interaction, birds fed low and medium DLYS:ME had a similar body weight gain being the heaviest while birds low DLYS:ME were the lightest. By an interaction (P < 0.05), the highest overall FCR value was observed for birds fed LD × low DLYS:ME and improved linearly when DLYS:ME increased to the highest level reaching a limit for birds fed HD × medium DLYS:ME. Calorie conversion linearly decreased (P < 0.001) with increments in DLYS:ME. Jejunal and ileal starch and protein digestibility were affected on d 21 and 42 but not on d 7 of age. Given the independence of response on BW and feed consumption, the use of BND as a flexible system in diet formulations has the potential to enable more accurate formulation for optimisation of growth performance of broiler chickens.

Enzymatic programs for broilers

Two trials were conducted on floor pens to evaluate the performance of broilers fed with diets contained different carbohydrases enzyme programs at different ages. Trial 1- The body weight gain (BWG) was affected by the Negative Control (NC) treatment (P<0.05). However, when the birds were fed with the NC diet + Amilase (AMY)+Xilanase (XYL), the BWG improved and reached the same level as the Positive Control (PC). Trial 2- The birds received enzymes supplementation in the total phase and others only in the grower phase.The ME reduction by 120 kcal/kg increased the feed conversion ratio (FCR) (P<0.05) when compared to PC and none of the enzyme programs was able to recover the performance. It can be concluded that it is possible to supplement with AMY during the role period of broiler chicken life or use an enzymatic program with AMY or AMY+XYL only in the grower phase.

Para avaliar o desempenho de frangos de corte alimentados com rações com diferentes programas de enzimas carboidrases, foram realizados dois experimentos. No experimento 1 as aves receberam suplementação de enzimas apenas na fase de crescimento foi observado redução no ganho de peso (GP) daqueles alimentados com a ração Controle Negativo (CN) ou seja, ração com redução de 120 kcal/kg. No entanto, quando os frangos receberam a ração CN + Amilase (AMI) + Xilanase (XIL) o GP foi semelhante encontrado no Controle Positivo (CP). No Experimento 2, algumas aves tiveram enzimas em todo período de criação e outras tiveram apenas na fase de crescimento. O tratamento CN apresentou pior conversão alimentar que o CP. Pode-se concluir que é possível suplementar com AMI todo o período de criação das aves ou utilizar um programa enzimático com AMI ou AMI + XIL somente na fase de crescimento, sem prejuízo ao desempenho das aves.

Níveis vitamínicos para frangos de corte: [revisão] / Vitamin levels for broiler: [review]

As vitaminas são micronutrientes que participam de inúmeros processos metabólicos do organismo, sendo, portanto, essenciais para ótima saúde e desempenho do animal. Poucos trabalhos foram realizados nos últimos anos para estimar os melhores níveis de vitaminas para frangos, havendo grande variação entre os níveis empregados comercialmente. Sendo assim, o objetivo deste trabalho é apresentar os conceitos atuais na avaliação das exigências de vitaminas para frangos e os níveis sugeridos pela indústria e pelo meio científico. A maioria dos níveis recomendados pelo NRC (1994) foi baseada em trabalhos antigos, realizados em condições controladas e considerando os níveis mínimos para evitar sinais de deficiência, não avaliando o melhor desempenho nas condições de desafio encontradas a campo. Além disso, as linhagens atuais apresentam maior taxa de crescimento e produção, resultando em maior necessidade nutricional para expressar seu potencial genético. Atualmente, além dos índices zootécnicos, têm se avaliado outros parâmetros na determinação das exigências de vitaminas, tais como imunidade, bem-estar e características da carcaça. Os níveis vitamínicos empregados comercialmente são geralmente superiores aos preconizados pelo NRC (1994), sendo que os níveis das lipossolúveis podem superar até 25 vezes as recomendações e os das hidrossolúveis até cinco vezes. O uso de maiores níveis de vitaminas na dieta de frangos resulta em maior desempenho produtivo e qualidade da carne.

Vitamins are micronutrients that participe in many metabolic processes of the organism, being essential to animal health and performance. There has been little research conducted over the last years on estimating the best vitamin levels for broilers, having great variation on comercial levels. The objective of this study was to present the actual concepts of the evaluation of vitamin requirements for broiler and the levels recomendated by industry and research centers. The major levels recomendated by NRC (1994) were based on former studies, with controled conditions and considering minimal levels to avoid deficiency signs, not evaluating the best performance on the challenge conditions found in field. Furthermore, modern strains presents greater growth rate and production, resulting in higher nutritional requirements to express their genetical potential. Nowadays, besides zootecnical parameters, it had been evaluating other parameters to determine vitamin requirements, as imunity, well being and carcass characteristics. Comercial vitamin levels used are generally superior than the suggested by NRC (1994), and the liposolubles levels may be higher than 25 times the recomendations and hydrosolubles five times higher. The use of higher vitamin levels in brolier diet results in greater productive performance and meat quality.