Stimulation with monochromatic green light during incubation alters satellite cell mitotic activity and gene expression in relation to embryonic and posthatch muscle growth of broiler chickens

Previous studies showed that monochromatic green light stimuli during embryogenesis accelerated posthatch body weight (BW) and pectoral muscle growth of broilers. In this experiment, we further investigated the morphological and molecular basis of this phenomenon. Fertile broiler eggs (Arbor Acres, n=880) were pre-weighed and randomly assigned to 1 of the 2 incubation treatment groups: (1) dark condition (control group), and (2) monochromatic green light group (560 nm). The monochromatic lighting systems sourced from light-emitting diode lamps and were equalized at the intensity of 15 lx at eggshell level. The dark condition was set as a commercial control from day 1 until hatching. After hatch, 120 male 1-day-old chicks from each group were housed under incandescent white light with an intensity of 30 lx at bird-head level. No effects of light stimuli during embryogenesis on hatching time, hatchability, hatching weight and bird mortality during the feeding trial period were observed in the present study. Compared with the dark condition, the BW, pectoral muscle weight and myofiber cross-sectional areas were significantly greater on 7-day-old chicks incubated under green light. Green light also increased the satellite cell mitotic activity of pectoral muscle on 1- and 3-day-old birds. In addition, green light upregulated MyoD, myogenin and myostatin mRNA expression in late embryos and/ or newly hatched chicks. These data suggest that stimulation with monochromatic green light during incubation promote muscle growth by enhancing proliferation and differentiation of satellite cells in late embryonic and newly hatched stages. Higher expression of myostatin may ultimately help prevent excessive proliferation and differentiation of satellite cells in birds incubated under green light.     

Human-Friendly Light-Emitting Diode Source Stimulates Broiler Growth

Previous study and our laboratory have reported that short-wavelength (blue and green) light and combination stimulate broiler growth. However, short-wavelength stimuli could have negative effects on poultry husbandry workers. The present study was conducted to evaluate the effects of human-friendly yellow LED light, which is acceptable to humans and close to green light, on broiler growth. We also aimed to investigate the potential quantitative relationship between the wavelengths of light used for artificial illumination and growth parameters in broilers. After hatching, 360 female chicks (“Meihuang” were evenly divided into six lighting treatment groups: white LED strips (400–700 nm, WL); red LED strips (620 nm, RL); yellow LED strips (580 nm, YL); green LED strips (514 nm, GL); blue LED strips (455 nm, BL); and fluorescent strips (400–700 nm, FL). From 30 to 72 days of age, broilers reared under YL and GL were heavier than broilers treated with FL (P < 0.05). Broilers reared under YL obtained the similar growth parameters with the broilers reared under GL and BL (P > 0.05). Moreover, YL significantly improved feeding efficiency when compared with GL and BL at 45 and 60 days of age (P < 0.05). In addition, we found an age-dependent effect of light spectra on broiler growth and a quantitative relationship between LED light spectra (455 to 620 nm) and the live body weights of broilers. The wavelength of light (455 to 620 nm) was found to be negatively related (R ² = 0.876) to live body weight at an early stage of development, whereas the wavelength of light (455 to 620 nm) was found to be positively correlated with live body weight (R ² = 0.925) in older chickens. Our results demonstrated that human-friendly yellow LED light (YL), which is friendly to the human, can be applied to the broilers production.     

Effect of In Ovo Feeding of Folic Acid on Subsequent Growth Performance and Blood Constituents Levels in Broilers

This study designed to determine effect of in ovo feeding of folic acid on subsequent growth performance and blood constituents levels in broilers. A total of 1000 fertile broiler eggs were divided into four groups. Control group (1) received no injection. In group 2, eggs received in ovo feeding of distiller water (40 µg). Group 3 received in ovo feeding of folic acid (40 µg). Groups 4 and 5 were similar to Group 3, except eggs injected with 80 and 120 µg of folic acid. All eggs were incubated and after hatch chickens were randomly assigned into their experimental groups. On days 1 and 42 post-hatch, chicken randomly selected and blood constituents, carcass characteristics, food intake, body weight gain and food conversion ratio were determined. According to the results, no significant difference detected on hatchability rate of the in ovo injected eggs (P?>?0.05). Dose dependent increase observed in glucose and folic acid levels in chicken in ovo injected with folic acid on day 1 post hatch (P?=?0.001). Blood glucose, folic acid and phosphorous levels increased (P?=?0.001) while cholesterol, triglyceride, HDL and LDL, calcium and alkaline phosphatase decreased in ovo injected with folic acid on day 42 post hatch (P?=?0.001). Food conversion ratio increased by in ovo injection of the folic acid (P?=?0.001). These results suggest folic acid had positive effects in broiler chicken.     

Effects of in ovo feeding of l-arginine on breast muscle growth and protein deposition in post-hatch broilers

In ovo feeding (IOF) of l-arginine (Arg) can affect growth performance of broilers, but the response of IOF of Arg on breast muscle growth is unclear, and the mechanism involved in protein deposition remains unknown. Hense, this experiment was conducted to evaluate the effects of IOF of Arg on breast muscle growth and protein-deposited signalling in post-hatch broilers. A total of 720 fertile eggs were collected from 34-week-old Arbor Acres breeder hens and distributed to three treatments: (1) non-injected control group; (2) 7.5 g/l (w/v) NaCl diluent-injected control group; (3) 0.6 mg Arg/egg solution-injected group. At 17.5 days of incubation, fertile eggs were injected 0.6 ml solutions into the amnion of the injected groups. Upon hatching, 80 male chicks were randomly assigned to eight replicates of 10 birds each and fed ad libitum for 21 days. The results indicated that IOF of Arg increased relative breast muscle weight compared with those of control groups at hatch, 3-, 7- and 21-day post-hatch (P<0.05). In the Arg-injected group, the plasma total protein and albumen concentrations were higher at 7- and 21-day post-hatch than those of control groups (P<0.05). The alanine aminotransferase activity in Arg group was higher at hatch than that of control groups (P<0.05). The levels of triiodothyronine at four time points and thyroxine hormones at hatch, 7- and 21-day post-hatch in Arg group were higher than those of control groups (P<0.05). In addition, IOF of Arg increased the amino acid concentrations of breast muscle at hatch, 7- and 21-day post-hatch (P<0.05). In ovo feeding of Arg also enhanced mammalian target of rapamycin, ribosomal protein S6 kinase-1 and eIF4E-bindingprotein-1 messenger RNA expression levels at hatch compared with those of control groups (P<0.05). It was concluded that IOF of Arg treatment improved breast muscle growth, which might be associated with the enhancement of protein deposition.     

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.     

Effect of a photoperiodic green light programme during incubation on embryo development and hatch process

This study was conducted to evaluate the effect of a 12-h light, 12-h dark (12L : 12D) photoperiod of green light during day 1 to day 18 of incubation time, on embryo growth, hormone concentration and the hatch process. In the test group, monochromatic light was provided by a total of 204 green light-emitting diodes (522 nm) mounted in a frame which was placed above the top tray of eggs to give even spread of illumination. No light–dark cycle was used in the control group. Four batches of eggs (n=300/group per batch) from fertile Ross 308 broiler breeders were used in this experiment. The beak length and crown–rump length of embryos incubated under green light were significantly longer than that of control embryos at day 10 and day 12, respectively (P<0.01). Furthermore, green light-exposed embryos had a longer third toe length compared with control embryos at day 10, day 14 and day 17 (P=0.02). At group level (n=4 batches), light stimulation had no effect on chick weight and quality at take-off, the initiation of hatch and hatch window. However, the individual hatching time of the light exposure focal chicks (n=33) was 3.4 h earlier (P=0.49) than the control focal chicks (n=36) probably due to the change in melatonin rhythm of the light group. The results of this study indicate that green light accelerates embryo development and alters hatch-related hormones (thyroid and corticosterone), which may result in earlier hatching.     

Effects of in ovo feeding of creatine pyruvate on the hatchability,growth performance and energy status in embryos and broiler chickens

The effects of in ovo feeding (IOF) of creatine pyruvate (CrPyr) on the hatchability, growth performance and energy status of embryos and broilers (Arbor Acres) were investigated. Five treatments were arranged as non-injected treatment (Control), 0.6 ml physiological saline (0.75%) injected treatment (Saline), and IOF treatments injected with 0.6 ml physiological saline (0.75%) containing 3, 6 or 12 mg CrPyr (CrPyr₃, CrPyr₆ or CrPyr₁₂) into the amnion per fertile egg on day 17.5 of incubation. After hatching, 80 male chicks from each treatment with similar weight close to the average BW of their pooled group were selected and randomly assigned into eight replicates of 10 chicks each. The results showed that the hatchability was not affected among groups, whereas the hatching weight of broilers in CrPyr₁₂ was significantly higher than the control and saline groups (P<0.05). At 21 day post-hatch, the BWs of broilers in CrPyr₆ and CrPyr₁₂ were increased relative to the control and saline groups (P<0.05). Chickens in CrPyr₆ and CrPyr₁₂ exhibited higher BW gain and feed intake than the control and saline groups during 8 to 21 days post-hatch and the entire experiment period (P<0.05). Compared with the control and saline groups, the total and relative weight of pectoral muscle of embryos or chickens were greater in CrPyr₆ and CrPyr₁₂ at 19^th day of incubation (19 E), hatch, 3 and 21 days post-hatch (P<0.05). The concentrations of glucose and glycogen in liver were increased in CrPyr₆ and CrPyr₁₂ at 19 E and hatch (P<0.05). Neither glycogen nor glucose concentration in pectoral muscle was altered among treatments (P>0.05). Irrespective of dosage, the concentrations of creatine and phosphocreatine, and activities of creatine kinase in embryos were enhanced in CrPyr treatments at 19 E when compared with the control and saline groups (P<0.05). The activities of glucose-6-phosphatase in liver in CrPyr₆ and CrPyr₁₂ treatments were higher than the control and saline groups at 19 E (P<0.05). In conclusion, these results indicated that IOF of CrPyr, especially at the level of 12 mg/egg, could improve energy status of embryos and hatchlings, which was useful for enhancing hatching weight, BW and pectoral muscle weight until the end of the experiments at 21 days post-hatch in broilers.     

Effect of cyclical cold stress during embryonic development on aspects of physiological responses and HSP70 gene expression of chicks

The objective of the present study was to evaluate the effects of cyclical lower incubation temperature at different embryonic ages on the hatchability, body and organs weights, thyroid hormones, and liver HSP70 gene expression of newly hatched chicks. In a completely randomized design, fertile eggs of a broiler breeder (34 weeks of age) were assigned to three treatment groups with six replicates and 145 eggs per each. The treatment groups were as: control group (C) that eggs were incubated at 37.6 °C during the whole incubation period; incubation temperature was decreased to 36 °C for 3 h per day at embryonic age from 12 to 14 (T1); and incubation temperature was decreased to 36 °C for 3 h per day at embryonic age from 15 to 17 (T2). No significant difference was found among treatments for hatchability (P>0.05). There were no differences (P>0.05) among treatments for body weight and liver weight, while heart weight of chicks in T1 and T2 groups were significantly higher than the control group (P<0.05). There were no differences (P>0.05) among treatments for the levels of thyroid hormones, however, the levels of both hormones tended to increase in chicks exposed to cold stress (T1 and T2). Chicks in T2 group had higher liver HSP70 gene expression compared with those in T1 and the control group (P<0.05). Cold stress in both incubation periods had no significant effect on the plasma levels of aspartate aminotransferase and alanine aminotransferase. Treatments had no effect on the plasma levels of glucose, cholesterol and triglyceride. The results of this study suggest that cyclical lower incubation temperatures (36 °C) at the embryonic age from day 15–17 could induce the liver HSP70 gene expression, without negative effects on the hatchability and body weight of hatched chicks.     

The dark side of white hair? Artificial light irradiation reduces cortisol concentrations in white but not black hairs of cattle and pigs

Analysing hair cortisol concentrations (HCCs) is a minimally invasive way to retrospectively assess long-term stress, and its application in studies of animal welfare and stress has attracted considerable interest. However, not only stress-related effects but also hair-specific characteristics and external influences can affect HCCs and interfere with the interpretation of results. Thus, it was the aim of this study to investigate the impact of daylight and UV irradiation on cortisol concentrations in the hairs of pigs and cattle. We also examined whether a potential irradiation effect on HCCs depended on the colour of the hair. For this purpose, black and white hair samples from 18 Saddleback pigs and 18 Holstein Friesian cattle were exposed to artificial light (both visible and UV) and compared with control hair samples from the same animals kept in the dark. Exposure to artificial light significantly decreased HCCs in both pigs (P < 0.05) and cattle (P < 0.001), and hair colour had an influence on HCCs, with black hair showing higher cortisol levels than white hair (cattle: P < 0.001, pigs: P = 0.07). The interaction between light exposure and hair colour was significant in both pigs (P < 0.01) and cattle (P < 0.001), so light exposure reduced HCCs in porcine white hair but not black hair. In cattle, light-exposed white hair exhibited lower hair cortisol levels than control white hair or black hair. These results demonstrate that artificial light irradiation degrades hair cortisol or favours its elimination by structural changes of the hair matrix. However, this effect was only detectable in white hair, indicating that the melanin pigments in black hair absorbed radiation, thereby reducing the effects of photodegradation. Compared with other known influencing factors on HCCs, such as age and body region, the influence of light irradiation was relatively low in this in vitro experiment. However, further studies should investigate this influence under real-life animal conditions, such as outdoor and indoor housing.     

Lameness and its relationship with health and production measures in broiler chickens

The aim of this study was to explore lameness and the associations between lameness and health/production measures of animal welfare in commercial broiler production, using the Welfare Quality® protocol for broilers. A total of 50 flocks were included in the sample and farm visits were conducted for lameness scoring at a mean age of 28.9 days. The percentage of animals (n=7500) in the six different gait score (GS) categories were GS0: 2.53%, GS1: 44.19%, GS2: 33.84%, GS3: 16.32%, GS4: 2.36% and GS5: 0.53%. Production and other welfare data were collected for each flock after slaughter. Higher gait scores were associated with increased hock burn score (P<0.02), increased footpad dermatitis score (P<0.01), reduced bird cleanliness score (P<0.01) and peat litter (P<0.01). Although not statistically significant, there was a tendency for increased flock gait score being associated with wet litter (P=0.07). In addition, condemnations at postmortem inspection were associated with increasing gait scores (P<0.05), indicating that at least a portion of the lameness cases display pathological changes on the carcasses. In conclusion, 19%of the birds showed moderate-to-severe lameness, which was associated with several production or health and welfare observations including feather cleanliness and condemnations as unfit for human consumption at slaughter. Although stocking density and growth rate are already known key factors for lameness, associations of lameness with hock burns, footpad dermatitis and cleanliness of the birds suggest that a suboptimal physical environment (e.g. litter- and air quality) may be detrimental to leg health. Further studies are needed to explore these associations in more detail.     

In ovo injection with glycerol and insulin-like growth factor (IGF-I): hatchability,intestinal morphometry,performance, and carcass characteristics of broilers

ABSTRACT

The objective of this study was to evaluate the effects of in ovo injection with glycerol (GLY) and insulin-like growth factor (IGF-I) on hatchability, biochemical parameters, intestinal morphometry, performance, and carcass characteristics of broiler chickens. A total of 400 fertilised eggs were distributed into five experimental groups. The treatments were arranged as non-injected (control), saline solution injected (0.9% NaCl solution), GLY solution injected (10 nmol/ml), IGF-I solution injected (100 ng/ml), and GLY + IGF-I solution injected. At 17.5 d of incubation, 0.5 ml of each solution was injected into the amniotic fluid of each egg of the injected groups. The injection of different solutions did not influence the hatchability and incubation time of the eggs. Compared to intact eggs, IGF-I and IGF-I+ GLY increased (p < 0.01) the blood IGF-I at hatching. Higher hepatic glycogen was observed (p < 0.05) with GLY or IGF-I. The tested substances decreased (p = 0.02) the fructose 1,6-biphosfate phosphatase activity but did not affect glycaemia. No difference in performance was observed in the first week. Higher feed intake and weight gain with lower feed conversion ratio was obtained ( p  < 0.05) with IGF-I at 14 d. At 21 d, higher weight gain was obtained (p = 0.05) with IGF-I, GLY, IGF-I, and GLY + IGF-I, resulting (p < 0.01) in birds with greater weight gain at 35 and 42 d of age. GLY provided higher villus height in the ileum at hatching and at 7 d of age. The tested solutions increased the relative weight of the liver at hatching. At 42 d of age, no carcass characteristics were influenced. It is concluded that GLY and IGF-I, together or separately, can be used in the in ovo feeding to improve the post-hatch performance of broilers, without affecting hatchability and carcass composition.     

Effect of light intensity on broiler behaviour and diurnal rhythms

Light intensity manipulation is an important management tool affecting broiler behaviour and physiology but still there is a debate regarding the optimum level to be used in confinement barns. Two experiments were completed to study the impact of light intensity (1, 10, 20 and 40 lx) on behaviour and diurnal rhythms of broilers raised to 35 d of age. For each experiment, 950 Ross × Ross 308 chicks were housed per room with replication of individual light intensity treatment in two environmentally controlled rooms. Within each large room, a small pen with 25 male and 25 female chicks was used for recording behaviour. Data were analyzed as a randomized complete block design with experiment acting as a block. All chicks were provided with 40 lx intensity and 23 h light until shifting to treatment light intensity and 17 h daylength at 7 d of age. For each replicate, behaviour was recorded for a 24 h period, starting at 16 or 17 d of age. At 23 d of age, three birds per room were bled at the start, middle and end of light and dark periods for melatonin estimation using RIA. When summarized over the 24 and 17 h observation periods, birds exposed to a light intensity of 1 lx rested more (P = 0.01) and preened (P < 0.05) and foraged (P < 0.05) less in comparison to other light intensities. Birds from all treatments exhibited diurnal rhythms for feeding, resting, drinking, walking, standing, foraging and preening behaviours with little or no activity during the 7 h dark phase. The serum melatonin levels at the start, middle and end of light and dark phases were unaffected by light intensity (P > 0.1). In conclusion, light intensity ranges from 1 to 40 lx did not affect melatonin levels or behavioural diurnal rhythms, but birds exposed to a light intensity of 1 lx rested more and preened less, potentially indicating a reduced welfare state.     

Temperature alterations during embryogenesis have a sex-dependent influence on growth properties and muscle metabolism of day-old chicks and 35-day-old broilers

Broiler eggs were either incubated at 37.8°C during the whole incubation period (control), or at higher (38.8°C, group H) and lower temperatures (36.8°C, group L) from embryonic day (ED) 7 up to ED 10 (ED 7 to 10) or from ED 10 up to ED 13 (ED 10 to 13). Before and after this temperature treatment the eggs were incubated at 37.8°C. The day-old chicks were weighted, sexed and fed up to day 35. On days 1 and 35 samples were taken from the breast and leg muscles for analyzing of the mitochondrial respiratory activity (MRA) and from the breast muscles for analysis of the cross-sectional areas (CSA) and the glycogen phosphorylase (GP), phosphofructokinase (PFK), lactate dehydrogenase (LDH), citrate synthase (CS) and cytochrome oxidase (COX) activities. Statistical analysis showed that treatment (control, group H, group L), sex and their interaction, but not the treatment period (ED 7 to 10; ED 10 to 13), significantly influenced the results. Group H chicks had lower (P⩽0.05) body and heart weights but higher (P⩽0.05) liver weights, CSA values, leg MRA as well as PFK, LDH, CS, GP and COX activities compared with the group L chicks. The results of the control chicks differ (P⩽0.05) from those of the group H (body, heart weight, COX), the group L chicks (liver weight, PFK, LDH, CS, GP) or the birds of both other groups (CSA). The group H broiler had higher (P⩽0.05) body and leg weights as well as LDH, CS, COX and GP activities than the group L broilers. The BWs and the LDH and GP results of the control broiler differ (P⩽0.05) from those of both other groups or from the results of the group H (CS) and group L broiler (COX). Female broilers had lower (P⩽0.05) body, breast and leg weights, but higher (P⩽0.05) CSA, LDH, CS and GP activities than the male animals. Analysis of treatment×sex interaction showed that group H hens had higher (P⩽0.05) body and breast weights, LDH and GP activities compared with the group L hens, whereas in the male broiler no effect of the interaction could be found, except for the lower (P⩽0.05) CSA values in the group H than group L cocks. The treatment effects are probably due to altered embryonic activity and related molecular mechanisms. The sex-related differences in the broiler indicate that these alterations already occur in the embryos and chicks, but become significant with the sexual dimorphism after hatch.     

In ovo silver nanoparticle supplementation for improving the post-hatch immunity status of broiler chickens

Silver nanoparticles (AgNano) are known for their unique physical, chemical and biological properties, enabling cell penetration and anti-inflammatory response. In Experiment 1, the effect of an in ovo administration of AgNano (15 µg/egg; n = 360) at different incubation times (d 7 and d 18) on hatchability parameters was explored. In Experiment 2, post-hatch performance of broilers (42 d, n = 250) was studied after in ovo AgNano administration: Group T1 remained un-injected, Group T2 was the sham control and Groups T3, T4 and T5 were injected with 12.5, 25 and 50 µg AgNano, respectively, at 18 d of incubation. Chick weight, chick to egg weight ratio and hatchability as well average daily gain, average daily feed intake and feed conversion ratio were similar in all treatment groups. No variation was seen in the weight of thymus; however, the bursa and spleen weight was increased (p < 0.05) in Groups T4 and T5 in comparison to Group T1. The in vivo immune response to phytohaemagglutinin-P was increased in Group T3 in comparison to Groups T1 and T2 (p < 0.05), while the response to sheep red blood cells was increased in all AgNano-treated groups in comparison with Group T1 (p < 0.01). The expression of toll-like receptors 2 and 4 genes was up-regulated in AgNano groups in comparison with Groups T1 and T2 (p < 0.01). In summary, an in ovo supplementation of AgNano carried out at d 18 of incubation is effective and modulates the post-hatch immune response without affecting the hatchability, growth and other performance parameters in broilers.     

Effects of dietary selenium and vitamin E on immune response and biological blood parameters of broilers reared under thermoneutral or heat stress conditions

A study was conducted using 360 broiler chickens to evaluate the effects of dietary vitamin E (0, 125 and 250 mg/kg), selenium (Se, 0, 0.5 and 1 mg/kg), or their different combinations on immune response and blood biological parameters of broilers raised under either thermoneutral (TN, 23.9 °C constant) or heat stress (HS, 23.9 to 37 °C cycling) conditions. Humoral immunity was assessed by intravenous injection of 7 % sheep red blood cell (SRBC) followed by evaluation of serum for antibody titers in primary and secondary responses. Heterophil to lymphocyte (H/L) ratio also determined as an indicator of stress. Furthermore, at the end of the experiment, birds were bled for determination of some biological parameters. There was a significant reduction in body weight and feed intake, but the feed conversion ratio increased when the birds were exposed to HS (P?<?0.05). Body weight and feed intake were not influenced significantly by dietary vitamin E and Se (P?>?0.05), whereas feed conversion was improved significantly by 125 mg/kg vitamin E (P?<?0.05). The liver and lymphoid organ weights as well as IgM and IgG, antibody titers for primary and secondary antibody responses to SRBC were reduced significantly under HS (P?<?0.05). Heat stress also resulted in a significant increase in H/L ratio (P?<?0.05). Dietary vitamin E resulted in improvement of primary and secondary antibody responses both in TN and HS broilers (P?<?0.05). The HS birds also showed an improved antibody titer in secondary response with high concentration of Se (P?<?0.05). Vitamin E and Se had interactive effects on anti-SRBC titers; however, no consistent differences were found between dietary levels during the study. The H/L ratio decreased by feeding vitamin E at both levels either under HS or TN conditions (P?<?0.05). The serum concentrations of glucose, triglycerides, total cholesterol, and LDL-cholesterol were increased but serum HDL-cholesterol decreased in HS broilers (P?<?0.05).     

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 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 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.