Response of meat chickens to different sources of arginine in low-protein diets.

Arginine activity in broiler diets can be supplied by L-arginine (Arg), guanidinoacetic acid (GAA) and L-citrulline (Cit), all of which are commercially available. This study was conducted to assess the effects of Arg source and level on broiler performance, nutrient digestibility and carcass parameters. Day-old Ross 308 cockerels (n = 768) were assigned to one of eight dietary treatments using a completely randomized design: normal protein (NP), low protein deficient in Arg (LP) and LP with two levels of either Arg (0.238% and 0.476%), GAA (0.309% and 0.618%) or Cit (0.238 and 0.476%). The LP was 5 percentage points lower in protein level than the NP. Wheat, sorghum, soya bean meal, canola meal, and meat and bone meal-based diets were fed over three feeding phases to 6 replicate floor pens with 16 birds each. Compared to NP, birds fed LP had reduced feed intake (FI, p < 0.001), reduced body weight gain (BWG, p < 0.001) and increased feed conversion ratio (FCR, p < 0.001) from day 0 to day 35. Additions of Arg or Cit to the LP at both levels resulted in increased BWG and reduced FCR (p < 0.05). Birds fed LP with GAA added had lower FCR (p < 0.05) but not higher BWG (p > 0.05) compared with the LP observed from day 0 to day 35. Supplementation of Arg, Cit and the low level of GAA to LP resulted in increased carcass yield, bone length, diameter and ash (p < 0.05) but did not increase ileal energy or nitrogen digestibility (p > 0.05). The findings indicate that Cit is an efficacious source of Arg activity in Arg-deficient diets.

Upregulation of genes encoding digestive enzymes and nutrient transporters in the digestive system of broiler chickens by dietary supplementation of fiber and inclusion of coarse particle size corn.

BACKGROUND: Measures to improve bird performance have been sought due to the imminent phase out of in-feed antibiotics in poultry and continued demand for higher poultry feeding efficiency. Increasing grain particle size and dietary fibre may improve gizzard function, digestive efficiency and nutrient absorption. This study was conducted to evaluate the effect increased particle size of corn and inclusion of sugarcane bagasse (SB) on mRNA expression of genes encoding digestive enzymes and nutrient transporters in broilers. RESULTS: A total of 336 day-old Ross 308 males were assigned in a 2 × 2 factorial arrangement of treatments with corn particle size - coarse 3576 µm or fine 1113 µm geometric mean diameter, and SB - 0 or 2% inclusion. Feed conversion ratio (FCR), weight gain and feed intake were measured from d 0-10 and d 10-24. The relative gizzard weight and mRNA expression of genes encoding digestive enzymes and intestinal nutrient transporters were measured on d 24. During d 10-24, a particle size × SB interaction was observed for FCR (P < 0.01), where birds fed coarsely ground corn (CC) with 2% SB had lower FCR than those fed CC without SB. A particle size × SB interaction was observed for both expression of pepsinogen A and C (P < 0.01) which were negatively correlated with FCR on d 24. Addition of 2% SB upregulated pepsinogen A and C only in CC fed birds. Further, 2% SB also upregulated pancreatic amylase (AMY2A) and intestinal cationic amino acid transporter-1 (CAT1). Inclusion of dietary CC upregulated duodenal amino peptidase N (APN), jejunal alanine, serine, cysteine and threonine transporter-1 (ASCT1), and ileal peptide transporter-2 (PepT2). CONCLUSION: These results suggest that both SB and coarse particle size modulate expression of genes encoding important digestive enzymes and nutrient transporters and thus are directly related to bird performance. These findings provide insights into the combination effects of dietary fiber and particle size in the future management of broiler feeding.

Immune responses following experimental infection with Ascaridia galli and necrotic enteritis in broiler chickens.

Broilers commonly suffer from necrotic enteritis (NE). Other gastrointestinal infectious diseases affect poultry, including nematode infections which are considered a re-emerging disease in barn and free-range systems. The aim of this study was to characterize the immune response of broilers after artificial infection with NE and contrast these with responses to the nematode Ascaridia galli and determine whether immune parameters measured during the course of infection can be used to distinguish infected from uninfected birds. A total of 96 one-day-old male Ross 308 broiler chickens were used in this study. At 10 days of age, broilers were randomly assigned to one of the following treatment groups: control birds (n = 32), A. galli infected birds (n = 32), or NE infected birds (n = 32) and inoculated with the appropriate infective agents. The immune response of birds was monitored through evaluation of haematology parameters, acute phase protein production, and intraepithelial intestinal lymphocyte population changes at 11, 16, 20, and 32 days of age. T-helper cells (CD4+CD8-) increased significantly over time, and were significantly higher in A. galli and NE compared to day 10 controls. In conclusion, α-1 glycoprotein levels can distinguish birds with NE from other birds, including those infected with A. galli; also T-helper cell numbers can distinguish both NE and A. galli from uninfected birds and thirdly, 10 days post infection is the best time point to evaluate the bird's immune response for A. galli infections.

A Multifactorial Analysis of the Extent to Which Eimeria and Fishmeal Predispose Broiler Chickens to Necrotic Enteritis.

Necrotic enteritis (NE) is an important infectious disease in chickens. Predisposing factors play critical roles both in disease outbreaks in the field and in models for experimental induction of disease. Systematic manipulation and study of predisposing factors help to optimize methods for the experimental reproduction of disease. The nature of such factors may play a confounding role in challenge models and, therefore, warrant investigation to determine their importance in industry-relevant NE reproduction models. In the present study, we examined the roles of dietary fishmeal inclusion, Eimeria inoculation (E), and Clostridium perfringens challenge (C) on broiler growth performance and induction of NE infection. The results showed that E, preceding C, greatly increased the severity of NE induced in broiler chickens, but fishmeal addition played only a marginal role in the challenge model. Bird performance was significantly affected by all three factors during the 35-day experimental period. Fishmeal increased body weight, but statistically significant effects of fishmeal were not observed on feed conversion ratio (FCR) and feed intake. Both Eimeria and C. perfringens significantly reduced body weight gain and feed intake. E but not C led to significantly poorer FCR. These findings indicate that dietary fishmeal may be removed from the model to allow the performance results of challenged chicks to be equivalent to the performance of chicks in the field. In conclusion, the present study demonstrates that an NE challenge model without fishmeal is valid and removes bird performance bias in the model introduced by feeding high fishmeal diets, refining the model to facilitate the yield of more commercially relevant results.

Dietary acylated starch improves performance and gut health in necrotic enteritis challenged broilers.

Resistant starch has been reported to act as a protective agent against pathogenic organisms in the gut and to encourage the proliferation of beneficial organisms. This study examined the efficacy of acetylated high amylose maize starch (SA) and butyralated high-amylose maize starch (SB) in reducing the severity of necrotic enteritis (NE) in broilers under experimental challenge. A total of 720 one-day-old male Ross 308 chicks were assigned to 48 floor pens with a 2 × 4 factorial arrangement of treatments. Factors were a) challenge: no or yes; and b) feed additive: control, antibiotics (AB), SA, or SB. Birds were challenged with Eimeria and C. perfringens according to a previously reported protocol. On d 24 and 35, challenged birds had lower (P < 0.001) livability (LV), weight gain (WG), and feed intake (FI) compared to unchallenged birds. Challenged birds fed SA and SB had higher FI and WG at d 24 and 35 (P < 0.05) compared to birds fed the control diet, while being significantly lower than those fed AB. Unchallenged birds fed SA or SB had higher FI at d 24 and 35 compared to those fed the control diet (P < 0.05). Birds fed SB had increased (P < 0.001) jejunal villus height/crypt depth (VH:CD) ratios at d 15, increased ileal (P < 0.001) and caecal (P < 0.001) butyrate levels at d 15 and 24, and decreased (P < 0.01) caecal pH at d 15. Birds fed SA had increased (P < 0.001) ileal acetate content at d 24 and decreased (P < 0.01) caecal pH at d 15. These results demonstrated that dietary acylated starch improved WG in birds challenged with necrotic enteritis. Depending on the acid used, starch acylation also offers a degree of specificity in short chain fatty acid (SCFA) delivery to the lower intestinal tract which improves gut health.

Use of yeast cell wall extract as a tool to reduce the impact of necrotic enteritis in broilers.

The use of a yeast cell wall extract derived from Saccharomyces cerevisiae (Actigen(®)) has been proposed as an alternative to in-feed antibiotics. This experiment was conducted to investigate the efficacy of yeast cell extract as an alternative to zinc bacitracin or salinomycin using a necrotic enteritis challenge model. A feeding study was conducted using 480-day-old male Ross 308 chicks assigned to 48 floor pens. A 2 × 4 factorial arrangement of treatments was employed. The factors were: challenge (- or +) and feed additive (control, zinc bacitracin at 100/50 mg/kg, yeast cell wall extract at 400/800/200 mg/kg, or salinomycin at 60 mg/kg in starter, grower, and finisher, respectively). Diets based on wheat, sorghum, soybean meal, meat and bone meal, and canola meal were formulated according to the Ross 308 nutrient specifications. Birds were challenged using a previously established protocol (attenuated Eimeria spp oocysts) on d 9 and 10(8) to 10(9) Clostridium perfringens (type A strain EHE-NE18) on d 14 and 15). Challenged and unchallenged birds were partitioned to avoid cross contamination. Challenged birds had lower weight gain, feed intake and livability compared to unchallenged birds on d 24 and d 35 (P < 0.05). Birds given zinc bacitracin, yeast cell wall extract, or salinomycin had improved weight gain and livability when compared to control birds given no additives. Challenge × additive interactions were observed for feed intake and weight gain on d 24 and d 35 (P < 0.01). The additives all had a greater positive impact on feed intake, weight gain, and livability in challenged than unchallenged birds. All challenged birds showed higher necrotic enteritis lesion scores in the small intestine sections when compared to unchallenged birds (P < 0.01). Birds fed yeast cell wall extract exhibited increased villus height, decreased crypt depth, and increased villus:crypt ratio when challenged. Yeast cell wall extract, zinc bacitracin, and salinomycin were effective in preventing performance decline from necrotic enteritis in the current study. This study indicates that yeast cell wall extract has promise as a tool for controlling necrotic enteritis.

Effects of L-arginine, guanidinoacetic acid and L-citrulline supplementation in reduced-protein diets on bone morphology and mineralization of laying hens.

The alterations in feed ingredients and the nutrient matrix to produce reduced-protein diets may affect bone morphology and mineralization in laying hens. This study was implemented to determine the effects of L-arginine (Arg), guanidinoacetic acid (GAA), and L-citrulline (Cit) supplementation to Arg-deficient reduced-protein diets on bone morphology, strength, and mineralization status of laying hens. Individually housed Hy-Line Brown laying hens were evenly distributed to five dietary treatments with 25 replicates per treatment from 20 to 40 wk of age. Treatments consisted of a standard protein diet (17% crude protein, SP), a reduced-protein diet deficient in Arg (13% crude protein, RP), and RP supplemented with Arg (0.35% Arg, RP-Arg), GAA (0.46% GAA equivalent to 0.35% Arg, RP-GAA), or Cit (0.35% Cit equivalent to 0.35% Arg, RP-Cit) to reach the Arg level of SP diets. Birds fed the SP diet had similar bone weight, ash, length, width, Seedor index, breaking strength, and serum mineral concentration, but higher toe B level (P < 0.001) compared to those fed the RP diet at wk 40. Birds fed the SP diet consumed more but also excreted more K and B compared to those fed the RP diet (P < 0.01). Birds fed the SP diet had lower Cu digestibility (P = 0.01) and higher B retention (P < 0.01) compared to those offered the RP diet. Supplementation of Arg, GAA, and Cit to the RP diet increased relative femur weight and length (P < 0.001). Citrulline supplementation also increased relative tibia and femur ash, and Zn digestibility (P < 0.05). Supplementation of GAA to the RP diet decreased serum Ca, P, and Mg levels, decreased tibia Fe and Mg levels and toe Mg level, but increased Al, Fe, Zn, and Mn digestibility (P < 0.05). The current findings demonstrated the capacity of laying hens to adapt to low mineral intake by increasing mineral utilization. Overall, bone morphology and breaking strength, and serum mineral level in laying hens were not influenced by dietary CP levels. Dietary Arg, GAA, or Cit supplementation were effective in improving bone morphology and mineralization in laying hens fed Arg-deficient RP diets.

Feeding recycled food waste improved feed efficiency in laying hens from 24 to 43 weeks of age.

There is renewed interest in utilizing food waste as animal feed due to its potential benefits in reducing feed cost and environmental impact while improving global food security. This study was conducted to examine the efficacy of recycled food waste-based feed for laying hen performance, egg quality, and nutrient digestibility. Hy-Line Brown hens (n = 150) were randomly distributed to three dietary treatments with 50 replicate cages of a single bird per treatment from 24 to 43 weeks of age. The treatments were: a standard/control feed based on wheat, sorghum, and soybean meal; a recycled food waste based-feed; and a 50:50 blend of control and food waste based-feed. Hens offered the food waste-based diets had similar egg weight, hen day egg production, and egg mass, but lower feed intake and higher feed efficiency, compared to those fed the control diets (P < 0.001). Hens fed the food waste diets exhibited lower shell breaking strength and shell thickness at week 34, and higher yolk color score and higher fat digestibility compared to the control treatment at week 43 (P < 0.001). Thus, feeding the recycled food waste based-feed maintained egg production while improving feed efficiency compared to the control feed.

Abductive statistical methods improve the results of calibration curve bioassays: An example of determining zinc bioavailability in broiler chickens.

In this paper, we discuss the theory behind calibration curve experiments and their application to a zinc (Zn) bioavailability study with broiler chickens. Seven replicates of 16 male commercial broiler chicks were fed starter diets for 14 days. Six diets had different levels of a potential Zn source and one was a positive control with standard industry levels of Zn for comparison. Four commonly used methods of calculating bioavailability means and confidence intervals (CI) from a calibration curve (standard curve) experiment to estimate the bioavailability of a new zinc source in broiler chickens were compared. The methods compared were the following: 1) the Counter-Intuitive Method uses a multiple-range test to compare unknown test and standard samples; 2) the Intuitive Method uses standard linear regression and inverts the equation to predict Zn bioavailability for each replicate of test samples; 3) the Abductive Method uses Graybill's Equation, based on theory and observation, to estimate CI's; and 4) the Sophistic Method uses reverse regression, and calculates Zn bioavailability values directly from the equation. The Counter-Intuitive Method only gives information about which standards the test samples are, or are not, significantly different from respectively (average available Zn not predicted). The Intuitive Method ignores error about the standard curve and theoretically cannot estimate the CI directly ( X ¯ ± SEM  = 107.5 ± 15.8 mg Zn/kg). The Sophistic Method underestimates and overestimates the test sample mean values above and below the mean of the standards, respectively ( X ¯  = 96.6 mg Zn/kg). The Abductive Method has an advantage over the other methods: The mean prediction estimation is consistent with theory (107.5 ± 6.1 mg Zn/kg; X ¯ ± SEM ). When test or "unknown" samples are near the mean of the standard samples, the CI is smaller than when near the extremes of the calibration curve. When calibration curve error is small (R 2 > approximately 0.95), there is little advantage to using the Abductive Method, but when calibration curve error is larger, as in many bioassays with growing animals, the Abductive Method improves the accuracy of the CI calculations. The Abductive Method was used to demonstrate the influence of the number of replicate samples on experimental power and cost.

Sources and levels of copper affect liver copper profile, intestinal morphology and cecal microbiota population of broiler chickens fed wheat-soybean meal diets.

Super dosing copper (Cu) has long been used as an alternative to antibiotic growth-promoters in broiler chickens' diet to improve gut health. This study was designed to compare nutritional and growth-promoting levels of Cu hydroxychloride (CH) with CuSO4 on gut health bio-markers and liver mineral profile of broiler chickens. Ross 308 chicks (n = 864) were randomly assigned to eight treatments, as basal diet containing no supplemental Cu; the basal diet with 15 or 200 mg/kg Cu as CuSO4; or 15, 50, 100, 150 or 200 mg/kg Cu from CH. The highest liver Cu content was observed in birds fed the diets with 200 mg/kg CuSO4 (P < 0.01). Serum FITC-d concentration as the leaky gut marker, and liver malondialdehyde concentration were not affected. Copper level or source had no effect on cecal short chain fatty acid and the mRNA expression of five jejunal genes involved in gut integrity. Negative linear responses of Cu were observed on Lactobacillus (P = 0.032), Bacteroides (P = 0.033), and Enterobacteriaceae (P = 0.028) counts. The jejunal villus height increased in birds fed CH at 200 and 100 mg/kg (P < 0.05). Increasing Cu levels, linearly and quadratically (P < 0.001), increased Cu excretion.

Organic acid blends improve intestinal integrity, modulate short-chain fatty acids profiles and alter microbiota of broilers under necrotic enteritis challenge.

Controlling enteric diseases of broilers is crucial. Among many additives, organic acids (OA) and their blends are gaining attention to combat diseases in the post-antibiotic era. The current study evaluated the potentials of short-chain fatty acids (SCFA) and medium-chain fatty acids (MCFA) blends and/or phenolic compounds on intestinal integrity, intestinal pH, caecal microbiota, and caecal SCFA profiles of broilers under necrotic enteritis (NE) challenge. The additives used were: (A) a blend of SCFA, MCFA, and a phenolic compound (SMP), (B) a blend of free and buffered SCFA with MCFA (SMF), and (C) a blend of free and buffered SCFA with a high concentration of MCFA (SHM). A total of 1,404 male parental chicks of Ross 308 broilers were randomly allocated to 78 floor pens on hatching day with 6 treatments replicated 13 times with 18 birds per pen. The treatments were: UCC, unchallenged control; CHC, challenged control; BAC, challenged group plus zinc bacitracin; SMP, challenged group plus additive SMP; SMF, challenged group plus additive SMF; SHM, challenged group plus additive SHM. Birds were challenged with field-strain Eimeria spp. on d 9 and Clostridium perfringens on d 14. Birds challenged with NE increased fluorescein isothiocyanate dextran (FITC-d) concentration in serum, reduced acetate and butyrate concentrations, and increased Bacteroides and C. perfringens load in the caeca (P < 0.05). Birds fed additives decreased FITC-d from gut to serum, reduced Bacteroides (d 16, P < 0.05) and numerically reduced C. perfringens load compared to CHC group. Birds fed additive SHM had higher concentrations of acetate and butyrate (d 21, P < 0.05) than CHC group but were not different from SMP and SMF groups. All the additives exhibited similar intestinal protection against NE compared to the BAC group indicated by FITC-d concentration in serum, acetate, propionate and butyrate concentrations in the caeca, and caecal bacterial loads except for the C. perfringens (P > 0.05). The SMP group had a higher load compared to BAC (P < 0.05). These findings suggest the promising effects of OA blends as alternatives to BAC to ameliorate the impact of NE challenge of broilers as indicated by improved intestinal health.

Effects of L-arginine and L-citrulline supplementation in reduced protein diets for broilers under normal and cyclic warm temperature.

Heat stress causes significant economic losses in the broiler industry. Dietary supplementation of arginine (Arg) and citrulline (Cit) might increase the performance of broilers raised under warm temperature due to vasodilation effects. This study investigated the effects of L-Arg or L-Cit supplementation in broilers fed a reduced protein wheat-based diet deficient in Arg under thermoneutral (NT) and cyclic warm temperature (WT). Ross 308 cockerels (n = 720) were randomly allocated to 4 dietary treatments with 12 replicates of 15 birds per pen from d 7 to 21. The 4 treatments were: normal protein (NP), i.e., 22.3% and 20.9% crude protein in grower and finisher, respectively; reduced protein (RP), i.e., 2.5% lower protein and deficient in Arg; and RP supplemented with 0.28% Arg (RP-Arg) or 0.28% Cit (RP-Cit). A factorial arrangement of treatments was applied during the finisher phase (21 to 35 d). Factors were: diet (4 diets above); and temperature, NT (24 °C) or cyclic WT (33 ± 1 °C for 6 h per day) with 6 replicate pens per treatment. During 7 to 35 d and 21 to 35 d, the birds fed the RP diet had lower body weight gain (BWG) and higher FCR compared to the NP diet (P < 0.01). The addition of Arg or Cit to RP decreased FCR compared to RP (P < 0.01). During 21 to 35 d, the birds exposed to WT had lower feed intake (FI), lower BWG (P < 0.001) but similar FCR (P > 0.05) compared to birds exposed to NT. Diet by temperature interactions were not observed for performance parameters during the period of WT (P > 0.05). On d 35, the RP-fed birds had a lower yield of thigh and drumstick, higher fat pad, lower femur ash, and breaking strength but similar serum uric acid level and higher nitrogen digestibility on d 21 compared to those offered NP (P < 0.05). Supplementation of Arg or Cit to RP resulted in increased femur ash on d 35 (P < 0.05). Thus, feeding the NP diets is necessary to maintain growth performance in broilers regardless of the temperature conditions.

Peroxisome proliferator-activated receptor gamma upregulation and dietary fat levels in laying hens.

The present study was conducted to investigate the effect of feeding the different levels of the dietary fat on the expression of genes encoding proteins involving energy metabolism, oxidative phosphorylation, and lipid synthesis including peroxisome proliferator-activated receptor gamma (PPARγ) of laying hens in the intestine. Birds fed diets with 3 levels of fat, that is, low (LF), medium (MF), and high fat (HF) were reared from 22 to 42 wk of age. Jejunum tissue was collected at week 42 for gene expression analysis. Dietary fat content as ether extract, net energy to AME ratio, and CP content of 3 treatment groups were as follows: LF: 25, 0.735, 187 (g/kg, DM); MF: 61, 0.739, 185 (g/kg, DM); HF: 73, 0.752, 181 (g/kg, DM). The BW, fat pad weight (g), fat pad-to-BW ratio (%) was the same for all the treatments (P > 0·05). Birds fed a diet containing HF increased the AME daily intake per metabolic BW (BW0.75) (P < 0.05). The expression of jejunal PPARγ was increased in the birds fed MF than that fed LF (P < 0.05). Dietary fat level did not affect the expression of other genes: protein kinase AMP-activated noncatalytic subunit gamma 2, NADH dehydrogenase subunit 2, succinate dehydrogenase complex flavoprotein subunit A, ubiquinol-cytochrome c reductase Rieske iron-sulfur polypeptide 1, cytochrome c oxidase subunit III, ATP synthase subunit alpha, avian adenine nucleotide translocator, and acetyl-CoA carboxylase alpha (P > 0·05). The mitochondrial count per cell showed no difference among the 3 groups with different dietary treatments (P > 0·05). The results suggest that PPARγ may be important to the energy expenditure during nutrient absorption, digestion, and metabolism, and respiratory chain complexes, and other genes involving mitochondrial energy metabolism and lipogenesis may be less responsive to dietary treatment.

Response of laying hens to l-arginine, l-citrulline and guanidinoacetic acid supplementation in reduced protein diet.

A study was conducted with Hy-Line Brown laying hens to examine the effects of reduced protein diet, deficiency of arginine (Arg), and addition of crystalline Arg, citrulline (Cit) and guanidinoacetic acid (GAA) as substitutes for Arg. Hen performance, egg quality, serum uric acid, liver and reproductive organ weights, and energy and protein digestibility were measured using a completely randomized design with 5 treatments. Treatments were a standard diet (17% protein diet; SP), a reduced diet (13% protein diet deficient in Arg; RP) and RP with added Arg (0.35%, RP-Arg), GAA (0.46% equivalent to 0.35% Arg, RP-GAA) or Cit (0.35%, RP-Cit) to the level of SP. It was hypothesized that performance would decrease with Arg deficient RP diet and the addition of GAA or Cit in RP would allow birds to perform similar or greater than Arg-added RP treatment. The experiment was conducted from 20 to 39 wk of age but the treatment effect was seen only after 29 wk of age. The birds offered RP had reduced egg and albumin weights (P < 0.01), lower yolk color score (P < 0.01), lower protein intake and excretion (P < 0.01) than those offered SP. When Arg or Cit were added to RP to make them equivalent to SP, feed intake (FI) and egg production were not different than those of RP (P > 0.05). The birds offered RP-GAA decreased FI and egg production (P < 0.01) compared to those offered RP. The addition of Arg, Cit or GAA to the RP had no effect on egg quality parameters, protein and energy digestibilities (P > 0.05). However, birds offered the RP-Cit diet tended to have higher Haugh unit (P = 0.095) and lower shell breaking strength (P = 0.088) compared to all other treatments while those offered RP-GAA had higher energy digestibility (P < 0.05) than all other groups but RP. The limited performance response of hens fed RP with added Arg, GAA, or Cit may be due to deficiency of some other nutrients in RP such as phenylalanine, potassium or non-essential amino acids and other components of soybean meal in the diet.

Multi-carbohydrase enzymes improve feed energy in broiler diets containing standard or low crude protein.

This study evaluated the effect of multi-carbohydrase (MC) on energy and nitrogen (N) balance and gene expression in broilers fed diets with different crude protein (CP) contents. The study employed a 2 × 2 factorial arrangement of treatments. The factors were presence or absence of MC, and standard (SCP) or low (LCP) dietary CP concentration. A 3-phase feeding program was used, including starter (0 to 7 d), grower (8 to 17 d) and finisher (18 to 28 d) phases. The study was undertaken in closed calorimetry chambers. Each of the 4 dietary treatments was replicated 8 times in total across 2 runs, with 2 birds per replicate (n = 64). Data for energy partitioning and N balance were collected from d 25 to 28. On d 28, birds were euthanized to collect muscle and intestinal tissue samples for gene expression. The results showed that the MC increased apparent metabolizable energy (AME, P < 0.01) and net energy (NE, P < 0.05), and reduced the feed conversion ratio (FCR, P < 0.01) in all diets. The proportion of energy retained as fat per total energy retention (REf/RE) was positively correlated with feed AME and NE (r = 0.541, P < 0.01 and r = 0.665, P < 0.001, respectively), suggesting that feed energy augmented with increased fat gain. Muscle ATP synthase subunit alpha (ATP5A1W) gene expression had a positive correlation with REf/RE and feed NE (r = 0.587, P < 0.001 and r = 0.430, P < 0.05, respectively). Similarly, muscle peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1A) expression was negatively correlated with weight gain and positively correlated with FCR (r = -0.451, P < 0.05 and r = 0.359, P < 0.05, respectively). These correlations show that over-expressions of muscle genes related to energy production reduce bird performance. This study demonstrated that MC increase dietary energy utilization, regardless of dietary CP concentration. However, the energy released by the enzymes increases feed energy-to-CP ratio, meaning there is excess energy that is then deposited as body fat. This suggests that supplemental MC in broiler feeds is beneficial if diets are formulated to contain marginal energy levels.

Multi-carbohydrase effects on energy utilization depend on soluble non-starch polysaccharides-to-total non-starch polysaccharides in broiler diets.

Non-starch polysaccharides (NSP), especially in water-soluble form, are a common anti-nutritional factor in cereal-based poultry diets. Consequently, carbohydrases are applied to diets to combat the negative effects of NSP on bird performance and health, particularly when feeding viscous grains. This study investigated the effect of supplementing multi-carbohydrases (MC) to broiler diets containing either low (LS) or high (HS) soluble NSP (sNSP) to total NSP (tNSP) ratios on energy partitioning, nitrogen (N) balance, and performance. A 2 × 2 factorial arrangement of treatments (MC, no or yes; sNSP/tNSP, LS vs. HS) was applied, resulting in 4 dietary treatments, each replicated 8 times. These treatments were fed to Ross 308 broilers in closed-circuit indirect calorimetry chambers, with 2 birds (a male and a female) per replicate chamber (n = 64). The results showed that MC addition increased AME, net energy (NE), and AME/gross energy, regardless of sNSP/tNSP content (P < 0.01 for all). There was an MC × sNSP/tNSP interaction for feed intake (FI, P < 0.05), denoting that in the absence of MC, the HS-fed birds had lower FI than LS-fed birds, but this difference was eliminated when MC was present. There were MC × sNSP/tNSP interactions observed for AME intake (AMEi) per metabolic BW (BW0.70, P < 0.05), AMEi/N retention (Nr, P < 0.01), NE intake (NEi)/Nr (P < 0.05), retained energy (RE) as fat per total RE (REf/RE, P < 0.01), and N efficiency (Nr/N intake, P < 0.05). These interactions showed that MC application increased AMEi/BW0.70, AMEi/Nr, NEi/Nr, and REf/RE only in the HS-fed birds, and N efficiency only in the LS-fed broilers. This study demonstrated that MC application markedly increased feed energy utilization in all diets, and increased N efficiency in birds fed an LS diet.

Differential expression of intestinal genes in necrotic enteritis challenged broiler chickens with 2 different Clostridium perfringens strains.

The primary cause of necrotic enteritis (NE) disease in chickens is the NetB-positive Clostridium perfringens bacterium. Many factors are known to affect the severity of NE in the challenge models of broiler chickens, and one of these factors is the virulence of C. perfringens strain. This study was conducted to evaluate the effect of 2 pathogenic C. perfringens strains in a NE challenge model on gut health and mRNA expression of genes encoding apoptosis, tight junction, immunity, and nutrient transporters in broilers. Day-old Ross-308 male broilers (n = 468) were allocated in a 2 × 3 factorial arrangement of treatments with in-feed antibiotics (no or yes) and challenge (Non, C. perfringens strain NE18, and C. perfringens strain NE36) as the factors. The birds in the challenged groups were inoculated with Eimeria species on day 9 and with a fresh suspension of C. perfringens NE18 or NE36 on day 14 and 15. Sample collection was performed on 2 birds of each pen on day 16. Necrotic enteritis challenge, impaired feed conversion ratio during day 0 to 16 compared with the control group where the effect of the NE36 challenge was more severe than that with NE18 (P < 0.001). The mRNA expression of mucin-2, immunoglobulin-G, occludin (P < 0.001), and tight junction protein-1 (P < 0.05) genes were downregulated in both challenged groups compared with the nonchallenged counterparts. Antibiotic supplementation, on the other hand, increased weight gain, and feed intake in all challenged birds (P < 0.01), but upregulated mucin-5ac and alanine, serine, cysteine, and threonine transporter-1 (P < 0.05) only in the NE18 challenged birds. The challenge with NE36 significantly upregulated caspase-8 and claudin-1 (P < 0.001), but downregulated glucose transporter-2 (P < 0.001) compared with the NE18 challenge. These results suggest that NE challenge is detrimental to the performance of broilers through compromised intestinal health, and different C. perfringens strains can affect the severity of the disease through modulating the expression of intestinal genes encoding proteins responsible for apoptosis, gut integrity, immunity, mucus production, and nutrient transporters.

Over-processed meat and bone meal and phytase effects on broilers challenged with subclinical necrotic enteritis: Part 3. Bone mineralization and litter quality.

This study was conducted to determine the effect of necrotic enteritis (NE), phytase level and meat and bone meal (MBM) processing on bone mineralization of broilers and litter quality. Ross 308 male broiler chicks (n = 768) were allotted to 48 pens with 16 birds each. There were 8 dietary treatments in a 2 × 2 × 2 factorial arrangement. Factors were NE challenge (no or yes), phytase level (500 or 5,000 FTU/kg), and MBM (as-received or over-processed). Half of the birds were challenged with field strains of Eimeria spp. at d 9 and 108 CFU per mL of Clostridium perfringens strain EHE-NE18 on d 14 and 15. The middle toe, tibia and femur of 2 birds per pen were excised at d 16 and 29 for determination of ash, breaking strength (BS) and bone mineralization. At d 42, all were assessed for hock burns and litter was scored and assessed for dry matter (DM). At d 16, challenged birds had lower toe ash (P < 0.01), femur ash (P < 0.001), tibia ash (P < 0.001) and tibial BS (P < 0.001) than unchallenged birds. At d 16, challenged birds fed high phytase and over-processed MBM had higher toe Mn than those fed low phytase and as-received MBM. At d 29 unchallenged birds fed high phytase and as-received MBM had a higher toe Mn than those fed over-processed MBM. At d 16, a phytase × MBM interaction was detected for femur Zn concentration (P < 0.05), where a higher level of Zn was observed in the high phytase group fed over-processed MBM. At d 16, tibial Ca (P < 0.05) and P (P < 0.05) were lower in the challenged whereas the femur K (P < 0.001), Mn (P < 0.01) and Na (P < 0.001) were higher in the challenged at d 16. At d 42, challenged birds had higher litter DM (P = 0.058) and fewer hock burns than those unchallenged (P < 0.05). In conclusion, NE impaired bone traits but high phytase and over-processed MBM increased bone mineral contents. Cases of hock burns may be lower under NE incidences due to lower livability of birds reducing litter wetness.

Potential of blended organic acids to improve performance and health of broilers infected with necrotic enteritis.

Organic acids (OA) and their blends have been shown to positively affect performance and health of broilers. However, the data in the literature are not consistent. This study examined the potential of blended short-chain fatty acids (SCFA) with medium-chain fatty acids (MCFA) as alternatives to antibiotic growth promoters (AGP) on performance, health and welfare of broilers infected with necrotic enteritis (NE). The additives used were: A) a blend of SCFA, MCFA, and a phenolic compound (SMP); B) a blend of free and buffered SCFA with MCFA (SMF); C) a blend of free and buffered SCFA with a high concentration of MCFA (SHM). A total of 1,404 Ross 308 one-day-old male parental chicks were randomly distributed into 78-floor pens with 13 replicates of 18 birds each. Six treatments were the following: T1, unchallenged control (UCC); T2, challenged control (CHC); T3, challenged group plus zinc bacitracin (BAC); T4, challenged group plus additive SMP; T5, challenged group plus additive SMF; T6, challenged group plus additive SHM. Challenged birds were gavaged with Eimeria spp. on d 9 and Clostridium perfringens EHE-NE18 on d 14. Post NE challenge and cumulatively, BWG, FCR, and nutrient digestibility of birds were compromised (P < 0.05) by NE challenge indicating a successful induction of sub-clinical NE. Additive SHM had higher BWG compared to CHC and BAC groups (P = 0.001; d 10 to 24) but not different from SMP and SMF groups (P > 0.05). All the 3 additive groups had lower FCR compared to CHC (P = 0.001; d 0 to 35), and exhibited similar jejunal lesions (d 16) compared to BAC and apparent ileal protein digestibility (d 21) compared to UCC and BAC groups (P > 0.05). Birds in additive SHM group had a higher concentration of serum IgA compared to all groups (P = 0.001) except additive SMF (P > 0.05; d 21). All the additive groups had lower footpad dermatitis and hock burns compared to CHC (P < 0.05). The findings suggest the potential of blended OA as alternatives to BAC to protect broilers from NE indicated by improved FCR, immunity, digestibility, and bird welfare.

Bacillus amyloliquefaciens CECT 5940 improves performance and gut function in broilers fed different levels of protein and/or under necrotic enteritis challenge.

Two studies were conducted to investigate the effect of Bacillus amyloliquefaciens CECT 5940 (BA) as a probiotic on growth performance, amino acid digestibility and bacteria population in broiler chickens under a subclinical necrotic enteritis (NE) challenge and/or fed diets with different levels of crude protein (CP). Both studies consisted of a 2 × 2 factorial arrangement of treatments with 480 Ross 308 mix-sexed broiler chickens. In study 1, treatments included 1) NE challenge (+/-), and 2) BA (1.0 × 106 CFU/g of feed) supplementation (+/-). In study 2, all birds were under NE challenge, and treatments were 1) CP level (Standard/Reduced [2% less than standard]) and 2) BA (1.0 × 106 CFU/g of feed) supplementation (+/-). After inducing NE infection, blood samples were taken on d 16 for uric acid evaluation, and cecal samples were collected for bacterial enumeration. In both studies, ileal digesta was collected on d 35 for nutrient digestibility evaluation. In study 1, the NE challenge reduced body weight gain (BWG), supressed feed conversion ratio (FCR) and serum uric acid levels (P < 0.001). Supplementation of BA increased BWG (P < 0.001) and reduced FCR (P = 0.043) across dietary treatments, regardless of challenge. Bacillus (P = 0.030) and Ruminococcus (P = 0.029) genomic DNA copy numbers and concentration of butyrate (P = 0.017) were higher in birds fed the diets supplemented with BA. In study 2, reduced protein (RCP) diets decreased BWG (P = 0.010) and uric acid levels in serum (P < 0.001). Supplementation of BA improved BWG (P = 0.001) and FCR (P = 0.005) and increased Ruminococcus numbers (P = 0.018) and butyrate concentration (P = 0.033) in the ceca, regardless of dietary CP level. Further, addition of BA reduced Clostridium perfringens numbers only in birds fed with RCP diets (P = 0.039). At d 35, BA supplemented diets showed higher apparent ileal digestibility of cystine (P = 0.013), valine (P = 0.020), and lysine (P = 0.014). In conclusion, this study suggests positive effects of BA supplementation in broiler diets via modulating gut microflora and improving nutrient uptake.