Modulation of intestinal mucin composition and mucosal morphology by dietary phytogenic inclusion level in broilers

The effect of a dietary phytogenic feed additive (PFA) inclusion level in mucin monosaccharide composition, mucosal morphometry and mucus histochemistry along the broiler intestinal tract was studied. Cobb male broilers (n = 525) were allocated into five experimental treatments that, depending on the type of addition in the basal diet (BD), were labeled as follows: C (BD based on maize–soybean meal with no other additions), E1 (80 mg PFA/kg BD), E2 (125 mg PFA/kg BD), E3 (250 mg PFA/kg of BD) and A (2.5 mg avilamycin/kg BD). Samples from duodenum, ileum and cecum of 14- and 42-day-old broilers were collected and analyzed. In 14-day-old broilers, treatments E2 and E3 had higher (P < 0.01) duodenal mannose than treatments C, E1 and A. Ileal mannose was lower (P < 0.05) in treatment C compared with PFA treatments, and ileal galactose (Gal) was higher (P < 0.01) in treatments E2 and E3 compared with C and A. Polynomial contrast analysis with respect to PFA inclusion level showed that in 14-day-old broilers there was a linear increase (P = 0.001) in duodenal mannose and a quadratic effect (P = 0.038) in duodenal N-acetyl-galactosamine with increasing PFA level. Ileal Gal and mannose increased linearly (P = 0.002 and P = 0.012, respectively) with PFA inclusion level. There were no significant differences between treatments in mucin monosaccharide molar ratios of 42-day-old broilers. However, increasing PFA inclusion level resulted in a linear decrease of ileal fucose (P = 0.021) and cecal N-acetylgalactosamine (P = 0.036). Experimental treatments did not differ (P > 0.05) regarding duodenal villus height (Vh), crypt depth (Cd) and Vh/Cd ratio, irrespective of broiler age and the intestinal segment examined. However, increasing dietary PFA inclusion level showed a pattern of linear increase of duodenal Vh/Cd ratio in 14-day-old broilers and ileal Vh in 42-day-old broilers (P = 0.039 and P = 0.039, respectively). Alcian Blue–Periodic Acid-Schiff (pH 2.5) staining of neutral and acidic mucins showed that the staining intensity of mucus layer in villi was fragment (i.e. tip, midsection and base) dependent, whereas in crypts it was dependent both on intestinal segment (i.e. duodenum, ileum and cecum) and fragment. Finally, mucus layer thickness did not differ (P > 0.05) between treatments, yet a pattern of linear increase (P < 0.05) with PFA inclusion level was observed in the duodenum of 42-day-old broilers. In conclusion, the dietary inclusion level of PFA modulated broiler intestinal mucin composition and morphology. Further studies are required to elucidate the physiological implications of such changes in host–microflora interactions.     

Effects of 5-aminolevulinic acid on the inflammatory responses and antioxidative capacity in broiler chickens challenged with lipopolysaccharide

5-Aminolevulinic acid (5-ALA) is an intermediate in haem biosynthesis and has anti-apoptotic, anti-inflammatory, antioxidant, and other pharmacological effects. This study aimed to investigate the effect of dietary supplementation with 5-ALA on growth performance, antioxidant capacity, and inflammatory response of the lipopolysaccharide (LPS)-challenged broiler chickens. The experiment was designed as a 2 × 2 factorial arrangement with dietary 5-ALA (0 or 60 mg/kg) and LPS (injection of saline or 0.5 mg/kg BW) levels as treatments. A total of 240 one-day-old Arbor Acres broilers were distributed into four treatments consisting of six replicates of 10 birds. All the experimental broilers were intraperitoneally injected with LPS or sterile saline at 16, 18, and 20 days of age. Our results showed that dietary 5-ALA supplementation reduced (P < 0.05) the feed to gain before broilers were stimulated with LPS (days 1–15). LPS challenge decreased (P < 0.05) the catalase (CAT), total superoxide dismutase activities and increased the content of malondialdehyde (MDA) in the serum of broiler chickens. However, 5-ALA supplementation had a tendency to increase (P = 0.08) the activity of CAT and decreased (P < 0.05) the content of MDA. LPS challenge showed higher (P < 0.05) interleukin (IL)-1β, IL-6, and IL-10 concentrations in the serum, whereas dietary 5-ALA supplementation decreased (P < 0.05) the levels of IL-1β and IL-6. Additionally, dietary 5-ALA supplementation significantly attenuated (P < 0.05) the upregulation of mRNA expression levels of hepatic toll-like receptor 4 (TLR4), IL-1β, and IL-2 induced by LPS challenge. Moreover, dietary 5-ALA supplementation also enhanced the mRNA expression of 5-aminolevulinate dehydratase, ferrochelatase, and haem oxygenase-1 (HO-1) as compared to the unsupplemented groups. In conclusion, our results suggested that supplementation of 60 mg/kg 5-ALA exhibited LPS-induced anti-inflammatory and antioxidant properties by enhancing the HO-1 expression and inhibiting the TLR4/NF-κB signalling pathway.     

Effects of Iron Glycine Chelate on Growth, Tissue Mineral Concentrations, Fecal Mineral Excretion, and Liver Antioxidant Enzyme Activities in Broilers

The study was conducted to determine the effects of iron glycine chelate (Fe-Gly) on growth, tissue mineral concentrations, fecal mineral excretion, and liver antioxidant enzyme activities in broilers. A total of 360 1-day-old commercial broilers (Ross?×?Ross) were randomly allotted to six dietary treatments with six replications of ten chicks per replicate. Broilers were fed a control diet with no Fe supplementation, while five other treatments consisted of 40, 80, 120, and 160?mg Fe/kg diets from Fe-Gly, and 160?mg Fe/kg from ferrous sulfate, respectively. After a 42-day feeding trial, the results showed that 120 and 160?mg Fe/kg as Fe-Gly improved the average daily gain (P?<?0.05) and average daily feed intake (P?<?0.05) of broilers (4?C6?weeks). Addition with 120 and 160?mg Fe/kg from Fe-Gly and 160?mg Fe/kg from FeSO₄ increased Fe concentration in serum (P?<?0.05), liver (P?<?0.05), breast muscle (P?<?0.05), tibia (P?<?0.05), and feces (P?<?0.01) at 21 and 42?days. There were linear responses to the addition of Fe-Gly from 0 to 160?mg/kg Fe on Fe concentration in serum (21?days, P?=?0.005; 42?days, P?=?0.001), liver (P?=?0.001), breast muscle (P?=?0.001), tibia (P?=?0.001), and feces (21?days, P?=?0.011; 42?days, P?=?0.032). Liver Cu/Zn superoxide dismutase activities of chicks were increased by the addition of 80, 120, and 160?mg Fe/kg as Fe-Gly to diets at 42?days. There were no differences in liver catalase activities of chicks among the treatments (P?>?0.05). This study indicates that addition with Fe-Gly could improve growth performance and iron tissue storage and improves the antioxidant status of broiler chickens.     

Effect of different levels of crude glycerol on the morphology and some pathogenic bacteria of the small intestine in male broilers

The gut health of poultry is closely associated with feeds and feeding. The experiment was conducted to the effect of crude glycerol addition to diets of male broilers on the bacterial microflora and morphology of their small intestines (duodenum, jejunum and ileum). A total of 120 Ross 308 broiler chicks received diets containing 0% (GLY 0), 4% (GLY 4) or 8% (GLY 8) crude glycerol for 42 days. The presence of Coliform bacteria and Enterobacteria was reduced in the duodenal tract of the broilers of GLY 4 (P<0.001); however, the presence of Staphylococci/Micrococci in the GLY 8 was reduced relative to the other groups (P<0.001). The presence of Salmonella spp. decreased in conjunction with the increasing quantities of glycerol (P<0.001). Analysis of the data regarding gut morphology (epithelial cell thickness, villi length and width, and crypt length and width) indicated that the glycerol levels fed to the different groups of broilers represented statistically different results in the small intestine. In general, whereas the diet with 4% glycerol statistically affected the investigated parameters of the gut, the diet with 8% glycerol statistically affected some segments of the broiler intestines.     

Effect of cereal type and plant extract addition on the growth performance,intestinal morphology,caecal microflora,and gut barriers gene expression of broiler chickens

Feeding broiler chickens on diets based on cereal grains of high non-starch polysaccharides content such as wheat and barley can negatively impact their performance and gut health. Plant extracts can be used as a potential tool to alleviate these negative effects. The present study assessed the effects of dietary cereal type and the inclusion of a plant extract blend (PEB) on the growth performance, intestinal histomorphology, caecal microflora, and gene expression of selected biomarkers for gut integrity in broiler chickens in a 42-d experiment. Ross-308 male broilers were assigned into different dietary treatments and fed on two cereal types (corn- vs. wheat/barley-based) with/without added graded concentrations of a PEB (0, 250, 500, 1000, and 2000 mg/kg diet). There were no significant differences in the growth performance parameters, intestinal histomorphology, and caecal microflora due to the impact of dietary cereal type. However, lactobacilli count in the caecal microflora was increased in the group fed on a corn-based diet. The PEB supplementation especially at a level of 500 to 1000 mg/kg diet significantly increased the average BW and decreased the feed conversion ratio. It also increased the villi length of duodenum, jejunum, and ileum, decreased the duodenal crypt depth, and increased the villi length to crypt depth ratio in the duodenum, jejunum and ileum. Supplementation of the PEB decreased the total bacterial and coliform count and increased the lactobacilli count in a linear pattern. Gene expression of Occludin and Junction Adhesion Molecule was significantly increased in the PEB supplemented diets, whereby no influence was observed on mucin expression. In conclusion, supplementation of a PEB at levels of 500–1000 mg/kg can be used as a tool to improve broiler performance and gut health.     

Effect of raw and fermented grape seed on growth performance,antioxidant capacity,and cecal microflora in broiler chickens

Grape seed (GS) is a by-product of the fruit juice and wine industry with the potential to be an alternative to synthetic antioxidants due to its antioxidant activity. Agro-industrial residues can be converted to more effective products by solid-state fermentation. The objective of the study was to investigate the effects of GS and fermented grape seed (FGS) on the growth performance, antioxidant capacity, and cecal microflora in broiler chickens. A total of 128 female broilers were randomly allocated into four treatment groups, each consisting of four replicates of eight birds. Throughout the 42-day feeding period, the birds were fed with soybean-corn based diet (CON), 0.15 g/kg synthetic antioxidant (butylated hydroxyanisole) supplemented diet (AO), 5 g/kg GS supplemented diet (GS), and 5 g/kg FGS supplemented diet (FGS). Dietary GS, FGS, and AO supplementation increased the BW (P < 0.05) and average daily weight gain (ADG, P < 0.05) compared with the CON group in the overall period of 42 days. Dietary FGS also increased the ADG (P < 0.05) in the period of 22–42 days compared with the control group. The pH of the breast meat of the chickens fed GS was higher (P < 0.01) than CON and FGS groups. Dietary FGS and AO decreased the b* value (P < 0.01) of breast meat compared with the CON group. Grape seed had the highest serum glutathione peroxidase (P < 0.05) and catalase (CAT, P < 0.01) levels among the treatment groups. The FGS also increased serum CAT level (P < 0.01) compared with the AO group. Moreover, dietary FGS supplementation increased Lactobacillus spp. (P < 0.05) in the cecum compared with the other treatment groups and decreased Staphylococcus aureus (P < 0.05) compared with the CON and AO groups. The present findings indicate that GS and FGS can be used in broiler diets as alternatives to synthetic antioxidants.     

Carbohydrases and phytase with rice bran,effects on amino acid digestibility and energy use in broiler chickens

Protein sources from cereals are used in broiler diets, usually in order to reduce feeding costs. However, their efficient use in poultry diets is limited by the level of fiber whose compounds are resistant to digestion in the small intestine; due to this sugars are not digested by endogenous poultry enzymes. The aim of this study was to determine the effect of multi-carbohydrase (MC) and phytase (Phy) on the total retention of nutrients, retention of apparent metabolizable energy corrected for nitrogen (AME_N) (trial 1) and apparent and standardized ileal digestibility of amino acids (trial 2) of rice bran (RB). A total of 245-day-old male broilers (Cobb 500) was distributed at 21-day-old in a completely randomized design in a 2 × 2 + 1 (0 and 200 mg/kg MC; 0 and 50 mg/kg Phy, and basal diet – BD) factorial arrangement of treatments, to give seven replicates and seven birds per replicate. The BD based on corn (trial 1) and cornstarch and casein (trial 2) was used only to determine the coefficients of retention of nutrients and energy, and coefficients of digestibility of amino acids of the RB. The test diets were made by mixing BD and RB 7 : 3 wt/wt basis. There was interaction (P<0.05) between MC × Phy for DM, nitrogen and AMEN, retention and no interaction (P>0.05) for ash, calcium, phosphorous and NDF was observed. Enzymes interacted (P<0.05) on standardized ileal digestibility of arginine, histidine, leucine, methionine, phenylalanine, threonine, valine, aspartic acid, glutamic acid, proline and serine. Dietary combination of MC and Phy resulted in higher (P<0.05) standardized digestibility of arginine, histidine, methionine and threonine relative to single enzyme supplementation or control diet without enzymes. Enzyme isolated inclusions in the diets improved (P<0.05) standardized digestibility of methionine. The supplementation of carbohydrases and Phy in RB will improve the nitrogen, energy and amino acids utilization for broiler chickens.     

In vivo immunomodulatory effects of plant flavonoids in lipopolysaccharide-challenged broilers

Plant flavonoids are generally regarded as natural replacers of synthetic growth promoters in poultry production. This study investigated the immunomodulatory effects of plant flavonoids, such as genistein and hesperidin, in lipopolysaccharide (LPS)-challenged broilers. A total of 700 21-day-old commercial Arbor Acres broiler chicks were randomly assigned into six treatment groups, each having six pens of 20 chicks/pen. Chicks were fed a basal diet without any additive (control, CON), 5 mg genistein/kg feed (G5), 20 mg hesperidin/kg (H20), or a basal diet with a combination of genistein and hesperidin (1 : 4) with doses of 5 mg/kg feed (GH5), 10 mg/kg (GH10) and 20 mg/kg (GH20) for 6 weeks. Half of the birds from each treatment were separated, and either challenged with 0·9% sodium chloride solution or Escherichia coli LPS (250 μg/kg BW) on days 16, 18 and 20. The results showed that both genistein and hesperidin improved (P<0.01) the plasma antioxidant status of growing broilers, by increasing total antioxidant capacity (TAOC), superoxide dismutase (SOD) activity and decreasing malondialdehyde production. LPS challenge further increased (P<0.05) TAOC and SOD levels. Regardless of LPS challenge, both genistein and hesperidin improved the humoral and mucosal immunity by increasing the intestinal intraepithelial lymphocyte numbers (P<0.01), as well as anti-Newcastle disease and anti-avian influenza antibody titers (P<0.05). Supplementation of both the plant flavonoids generally increased (P<0.05) the immune organs indices (spleen, thymus and bursa). Thus, supplementation of basal diet of broiler chicks, either with genistein or hesperidin, improved immune and antioxidant status of growing broilers. In addition, combined supplementation of both the flavonoids showed further improvement than individual compounds.     

Effects of feed form and energy levels on growth performance,carcass yield and nutrient digestibility in broilers

Feed form is well recognized to improve broiler performance, specially by increasing feed intake (FI). However, when different diet energy levels are used, the results differ in the literature. Therefore, this experiment was conducted to evaluate the influence of feed form and dietary metabolizable energy (ME) levels on broiler performance, carcass yield and on the digestibility of DM, CP, starch and gross energy. In total, 1152 male Cobb 500 broilers were evaluated between 35 and 47 days. The birds were distributed according to a completely randomized design in a 2 × 4 factorial arrangement, consisting of two feed forms (mash or pellet) and four ME levels (12.73, 13.06, 13.40 or 13.73 MJ/kg), totaling eight treatments with eight replicates of 18 birds. Broilers fed the lowest ME level presented the lowest weight gain (WG) and worst feed per unit gain (P < 0.01). Metabolizable energy intake increased (P < 0.01) with progressive increments of ME, which, however, did not affect caloric conversion (CC, P > 0.05). Pelleted diets promoted higher FI, WG, ME intake (P < 0.01) and better feed per unit gain and CC (P < 0.05) compared with mash. In mash diets, increasing dietary ME levels promoted a linear increase in WG (P < 0.01) and reduced feed per unit gain (P ≤ 0.05), but did not affect FI (P > 0.05). In pelleted diets, on the other hand, increasing ME levels linearly reduced FI (P < 0.05) and feed per unit gain (P < 0.01). Broilers fed pelleted diets presented higher abdominal fat deposition than those fed mash (P < 0.05). Increasing ME levels reduced the coefficients of ileal apparent digestibility of DM (P < 0.01) and total starch (P < 0.05) but did not affect the digestibility of other evaluated nutrients. The digestibility of all nutrients was lower when pelleted diets were fed compared with mash. Increasing inert material inclusion in the diets at the expense of soybean oil to reduce dietary ME levels promoted higher pellet durability index values (P < 0.05) and the percentage of fines (P < 0.01). Overall, the results suggest that pelleted diets promote better broiler performance because they increase FI, since the digestibility of dietary fractions is reduced. Chickens consuming low-energy pelleted diets may increase FI to compensate for energy deficit. In contrast, broilers fed mash diets may have reached their maximum intake capacity and did not regulate FI by changing feed energy density. When feeding pelleted diets, dietary energy reduction should be considered to reduce feed costs and to improve the carcass quality of broilers.     

Effects of dietary supplementation with creatine monohydrate during the finishing period on growth performance,carcass traits,meat quality and muscle glycolytic potential of broilers subjected to transport stress

A total of 320 male Arbor Acres broiler chickens (28 days old) were randomly allotted to one of the three experimental diets supplemented with 0 (160 birds), 600 (80 birds) or 1200 mg/kg (80 birds) creatine monohydrate (CMH) for 14 days. On the morning of 42 day, after an 8-h fast, the birds of CMH-free group were divided into two equal groups, and all birds of these four groups were transported according to the follow protocol: 0.75-h transport without CMH supplementation (as a lower stress control group), 3-h transport without CMH supplementation, 3-h transport with 600 or 1200 mg/kg CMH supplementation. Each treatment group was composed of 8 replicates with 10 birds each. The results showed that supplementation of CMH for 14 days before slaughter did not affect the overall growth performance and carcass traits of stressed broilers (P>0.05). A 3-h transport decreased plasma glucose concentration, elevated plasma corticosterone concentration, increased bird live weight loss, breakdown of muscle glycogen, as well as the accumulation of muscle lactate (P<0.05), which induced some detrimental changes to breast meat quality (lower ultimate pH and higher drip loss, P<0.05). Nevertheless, supplementation of 1200 mg/kg CMH reduced chicken weight loss, decreased the contents of lactate and glycolytic potential in pectoralis major of 3-h transported broilers (P<0.05), which is beneficial to maintain breast meat quality by reducing the drip loss (P<0.05). These findings suggest that the reduction of muscle glycolysis is probably the reason for maintainance of meat quality by supplementation of CMH in transported broilers.     

Effect of polyphenols extracted from tamarind (Tamarindus indica L.) seed coat on pathophysiological changes and red blood cell glutathione peroxidase activity in heat-stressed broilers

The purpose of this study was to determine the effect of polyphenols extracted from the tamarind seed coat (PETSC) on glutathione peroxidase (GPx) activity, red blood cell parameters and bilirubin in heat-stressed broilers. One hundred forty-seven broilers, 18-days old were divided into two groups. In group 1, broilers were maintained at an environmental temperature of 26?±?2 °C throughout the experimental period. In group 2, the broilers were maintained at 38?±?2 °C (cyclic temperature: 26?±?2 °C; ?38?±?2 °C; and ?26?±?2 °C, and broilers were maintained at 38?±?2 °C for 6 h/ day) and received PETSC at a concentration of 0, 100, 200, 300, 400 or 500 mg/kg in their diet ad libitum. Parameters were investigated on days 1, 7, 14 and 21 of the experimental period. Results showed that GPx activity of heat-stressed broilers that received 100 mg/kg of PETSC in their diet was lower (P?<?0.05) than that in broilers fed the other concentrations. The mean total red blood cell count and hemoglobin concentration of heat-stressed broilers that received 100 mg/kg PETSC was higher (P?<?0.05) than those in broilers in group 1 and those fed the other concentrations. The mean bilirubin level in the excreta of heat-stressed broilers that received 100 mg/kg of PETSC was lower (P?<?0.05) than that in broilers that received 0, 300, 400 and 500 mg/kg of PETSC. This showed that PETSC could reduce GPx activity and bilirubin in feces, and increase red blood cell parameters in heat-stressed broilers.     

Azolla leaf meal at 5% of the diet improves growth performance,intestinal morphology and p70S6K1 activation,and affects cecal microbiota in broiler chicken

With growing concern about including unconventional dietary protein sources in poultry diets to substitute the protein sources that are essential for human consumption such as soybean meal, Azolla leaf meal (ALM) has grown in popularity. In our prior experiment, ALM was used at inclusion rates of 5 and 10%. Five per cent inclusion of ALM increased broiler chicken growth performance, the concentration of cecal propionic acid, and activation of skeletal muscle p70S6 Kinase1 (p70S6K1) without having detrimental effects on the meat quality. Those results prompted us to further evaluate the effect of the same inclusion rates of ALM on phase feeding and intestine and liver health of the broiler chicks. The current study hypothesis is that dietary ALM positively affects phase feeding, intestinal morphology and p70S6K1 activation, cecal microbial gene expression, and improves the liver energy status. For this, we enrolled 135 one-day-old broiler chicks and collected growth performance data (starter, grower, and finisher stages) and samples of the gastrointestinal tract to analyse the morphology of the villi, immune-related organs, mucin, and abundance of intestinal p70S6K1. Cecal bacterial species were analysed using qPCR and liver samples were collected to analyse adenosine monophosphate (AMP) and ATP content and selected oxidative stress biomarkers. ALM increased BW and feed intake during the starter and grower phases but did not affect the feed conversion ratio. Liver oxidative stress and AMP: ATP ratio increased in chickens fed on a diet containing 10% ALM (AZ10; P < 0.05). Jejunum villi length and abundance of duodenal neutral mucin increased but villi of the ileum decreased in chickens fed on a diet containing 5% ALM (AZ5), while lymphoid follicle areas of the cecal tonsils decreased with both doses of ALM. Activation of p70S6K1 increased with AZ10 in the duodenum and AZ5 in the jejunum. In the gut, the family of Enterobacteriaceae decreased with both ALM doses. In conclusion, our results indicate an overall positive effect of dietary inclusion of ALM in the broiler chicken diet via its positive effect on intestinal morphology and function; however, a negative effect on the liver was observed with 10% ALM.     

How Bad Is Aluminum Exposure to Reproductive Parameters in Rats?

The objective of this study was to evaluate influence of dietary palygorskite (Pal) supplementation on growth performance, mineral accumulations in the tissues (livers, kidneys, and muscles), antioxidant capacities, and meat quality of broilers fed lead (Pb)-contaminated diet. One-hundred forty-four male broiler chicks were randomly divided into three treatment groups, receiving a corn-soybean meal basal diet (the control group), the basal diet contaminated with 10 mg/kg Pb (the Pb group), and the basal diet with 10-g/kg Pal supplementation and 10-mg/kg Pb contamination (the Pal/Pb group) from 1 to 42 days of age, respectively. Treatments did not affect growth performance of broilers in the 42-day study (P > 0.05). Compared with the control group, Pb contamination increased Pb accumulation in the livers, kidneys, and muscles (P < 0.05); elevated malondialdehyde accumulation in the livers, kidneys, and breast muscles; glutathione peroxidase activity in the livers and superoxide dismutase activity in the kidneys (P < 0.05); exacerbated drip loss in the pectoralis muscles (P < 0.05); and reduced glutathione peroxidase activity in the pectoralis muscles (P < 0.05) of broilers at 42 days of age. The values of these parameters were reversed in the Pal/Pb group to levels comparable with those in the control group (P < 0.05). Additionally, Pal supplementation reduced redness value in the pectoralis muscles (P < 0.05), and decreased Cu concentration in the pectoralis muscles and livers at 42 days of age as well as its accumulation in the kidneys at both 21 and 42 days of age compared with the other two groups (P < 0.05). The results suggested that dietary Pal supplementation would decrease Pb residue in the tissues, alleviate oxidative stress, and affect meat quality of broilers exposed to Pb.     

Effect of Dietary Supplementation with Copper Sulfate or Tribasic Copper Chloride on the Growth Performance, Liver Copper Concentrations of Broilers Fed in Floor Pens, and Stabilities of Vitamin E and Phytase in Feeds

An experiment was conducted using a total of 840, 1-day-old, Arbor Acres commercial male broilers to compare copper (Cu) sulfate and tribasic Cu chloride (TBCC, Cu₂(OH)₃Cl) as sources of supplemental Cu for broilers fed in floor pens. Chicks were randomly allotted to one of seven treatments for six replicate pens of 20 birds each, and were fed a basal corn–soybean meal diet (10.20 mg/kg Cu) supplemented with 0, 100, 150, or 200 mg/kg Cu from either Cu sulfate or TBCC for 21 days. Chicks fed 200 mg/kg Cu as TBCC had a higher (P?<?0.05) average daily gain (ADG) than those consuming other diets. Liver Cu contents of broilers fed diets supplemented with TBCC were numerically lower (P?>?0.05) than those of broilers fed diets supplemented with Cu sulfate. The vitamin E contents and the phytase activities in the feed fortified with TBCC were higher (P?<?0.01) and numerically higher (P?>?0.05) compared with those in the feeds fortified with Cu sulfate stored at room temperature, respectively. The vitamin E contents in liver and plasma of broilers fed diets supplemented with TBCC were higher (P?<?0.05) than those of birds fed diets supplemented with Cu sulfate. This result indicates that TBCC is more effective than Cu sulfate in improving the growth of broilers fed in floor pens, and it is chemically less active than Cu sulfate in promoting the undesirable oxidation of vitamin E in feeds.     

Soybean oligosaccharides attenuate odour compounds in excreta by modulating the caecal microbiota in broilers

Abatement of odour emissions in poultry production is important to ensure the quality and safety in the poultry industry as well as for the benefit of the environment. This study was conducted to compare the effects of supplementation with different amounts of dietary soybean oligosaccharides (SBO) and chlortetracycline (CHL) on the major odour-causing compounds in the excreta and on the caecal microbiota in broiler chickens. One-day-old broiler chickens were subjected to a 42-day experiment involving 6 treatments with 6 replicates per treatment (10 birds/cage). The treatments were as follows: negative control (NC; basal diet), positive control (PC; basal diet supplemented with CHL) and basal diet supplemented with 0.5, 2.0, 3.5 and 5.0 g/kg SBO (0.5SBO, 2.0SBO, 3.5SBO and 5.0SBO, respectively). Fresh excreta were sampled for the analysis of odour compounds by HPLC. Caecum content was collected for the caecal microbiota analysis through 16S rRNA sequencing. Results showed that on day 42, the excreta indole concentration in the broilers fed with 2.0SBO, 3.5SBO and 5.0SBO and PC was significantly decreased (P < 0.01) compared with that in the NC-fed broilers. The excreta skatole concentration (P < 0.001) and pH (P < 0.05) were also decreased by SBO and CHL, and they were lowest in birds fed with 3.5SBO. The formate concentrations in birds fed with 3.5SBO and 5.0SBO were higher than those in birds fed with the other diets (P < 0.001). Similarly, acetate concentration (P = 0.003) was increased in birds fed with 3.5SBO. Deep sequencing of 16S rRNA revealed that the composition of the caecal microbial digesta was slightly or significantly changed by the SBO or by the CHL supplementation, respectively. The SBO supplementation decreased the abundance of Bacteroides, Bilophila and Escherichia, which were associated with the skatole and indole concentrations in the excreta. By contrast, the CHL supplementation demonstrated a strong tendency to enrich Ruminococcus and to reduce the abundance of Rikenella. These results indicated that supplementation with dietary SBO is beneficial in attenuating the concentration of odour-causing compounds and that it modulates the composition of caecal microbiota in broiler chickens.     

Effects of Zinc Glycine Chelate on Oxidative Stress,Contents of Trace Elements,and Intestinal Morphology in Broilers

Three hundred and sixty healthy Ross × Ross 1-day-old broilers were used to study the effects of zinc glycine chelate (Zn-Gly) on oxidative stress, contents of trace elements, and intestinal morphology. All broilers were randomly assigned to six treatment groups, which replicates three times. Diets were as follows: (1) control (containing 29.3 mg zinc (Zn)/kg basic diet (0–21 days) and 27.8 mg Zn/kg (22–42 days)); (2) basic diet plus 30 mg Zn/kg from Zn-Gly; (3) basic diet plus 60 mg Zn/kg from Zn-Gly; (4) basic diet plus 90 mg Zn/kg from Zn-Gly; (5) basic diet plus 120 mg Zn/kg from Zn-Gly; and (6) positive control, basic diet plus 120 mg Zn/kg from zinc sulfate (ZnSO₄). The results showed that the addition of 90 or 120 mg/kg Zn-Gly led to an improvement of activity of Cu/Zn superoxide dismutase and glutathione peroxidase and a reduction of malondialdehyde content in livers at 21 and 42 days. With 90 mg/kg Zn-Gly, the content of sera zinc increased by 17.55% (P < 0.05) in 21-day broilers and 10.77% (P > 0.05) in 42-day broilers compared with that of the control. Adding 120 mg/kg Zn-Gly or ZnSO₄ to broilers' diets greatly enhanced the content of zinc in feces at 21 days (P < 0.05) and at 42 days (P < 0.05). For 42-day chickens, increased villus height and decreased crypt depth of the jejunum could be observed in the second growth stage of broilers fed with 90 mg/kg Zn-Gly. Also, intestinal wall thickness decreased (P < 0.05). In addition, adding 90 mg/kg Zn-Gly to the diet markedly elevated villus length of duodenum and decreased crypt depth of ileum (P < 0.05) in 42-day broilers.     

Effect of in ovo feeding of vitamin C on antioxidation and immune function of broiler chickens

Hypoimmunity and numerous stresses are two major challenges in broiler industry. Nutrient intervention at the specific time of embryonic stage is a feasible way to improve animal performance. This study was conducted to investigate the possible effects of in ovo feeding (IOF) of vitamin C at embryonic age 15^th day (E15) on growth performance, antioxidation and immune function of broilers. A total of 240 broiler fertile eggs were randomly divided into two groups (0 and 3 mg injected dose of vitamin C at E15), and new-hatched chicks from each treatment were randomly allocated into six replicates with 10 chicks per replicate after incubation. The results indicated that in ovo vitamin C injection improved the hatchability (P < 0.05) and increased immunoglobulin M (IgM) (at the broiler’s age 1^st day, D1), IgG and IgM concentrations (D21), as well as lysozyme activity (D21, P < 0.05) and total antioxidant capacity (D42, P < 0.01) in plasma of broilers. On D21, the splenic expression level of DNA methyltransferase 1 (DNMT1) was up-regulated in vitamin C (VC) group, whereas interleukin (IL)-6, interferon-γ, ten-eleven translocation protein 1 and thymine-DNA glycosylase were down-regulated (P < 0.05). On D42, in ovo vitamin C injection up-regulated splenic expression levels of DNMT1, DNA methyltransferase 3B (DNMT3B) and growth arrest and DNA-damage-inducible protein beta (P < 0.05), whereas down-regulated splenic expression levels of IL-6, tumour necrosis factor-α and methyl-CpG-binding domain protein 4 (P < 0.05). Our findings suggested that IOF of 3 mg vitamin C at E15 could improve, to some extent, the antioxidant activity and immune function in plasma, corresponding with the lower expression of pro-inflammatory cytokines in spleen. However, IOF of vitamin C leading to the changes in the expression of DNA methyltransferases and demethylases may suggest an increased trend of DNA methylation level in spleen and whether DNA methylation variation is associated with the lower expression of pro-inflammatory cytokines in spleen warrants future study.     

Effects of free and dipeptide forms of methionine supplementation on oxidative metabolism of broilers under high temperature

Our previous studies have shown that methionine supplementation could help to attenuate the effects of heat stress on the metabolism of broiler chickens. Here we investigated for the first time the effects of methionine supplementation in the form of DL-methionyl-DL-methionine on broilers subjected to heat stress during the growth phase. Broilers were divided into two groups; one group was reared under thermoneutral conditions and the other under continuous heat stress (30 ± 1 °C, 60% relative humidity). Both groups were subdivided into three dietary treatments: a methionine-deficient (MD) diet, a diet supplemented with free methionine (DL-M), and a diet supplemented with methionine dipeptide (DL-MM). Broilers raised under chronic heat stress had lower feed intake and weight gain than broilers raised under thermoneutral conditions (P < 0.05). There were no differences in animal performance between methionine-supplemented diets (DL-M and DL-MM). Heat-stressed birds had significantly higher heterophil/lymphocyte (H/L) ratio than thermoneutral birds. Under heat stress, broilers fed DL-M and DL-MM diets had lower H/L ratio than birds fed the MD diet. Higher concentrations of carbonylated proteins and lower concentration of reduced glutathione were observed in broilers raised under heat stress. In comparing heat-stressed broilers, we found that birds fed the DL-M diet had lower concentrations of thiobarbituric acid-reactive substances and carbonylated proteins than those fed the MD diet (P < 0.05). Higher expression of glutathione peroxidase (GPX) and glutathione synthetase (GSS) genes was observed in heat-stressed broilers (P < 0.05). Under heat stress, the MD diet increased GPX expression compared with other diets. Under thermoneutral conditions, the DL-M diet resulted in the highest GSS expression. There was a negative correlation between DNA methylation and GPX and GSS expression. Our results showed that supplementation of broiler diets with free methionine or methionine dipeptide may help attenuate the effects of heat stress through enhanced activation of genes related to the glutathione antioxidant system. Methionine effects were found for gene regulation, gene expression, and post-translational processing.