Modelling and optimizing of calcium and non-phytate phosphorus requirements of male broiler chickens from 1 to 21 days of age using response surface methodology

The skeleton is the main site of P and Ca deposition; therefore, accurate estimation of Ca and P requirements is necessary to maintaining health and optimum performance of broiler chickens. A response surface methodology (RSM) using a central composite design (CCD) was used for evaluating and optimizing of Ca and non-phytate P (NPP) requirements of broiler chickens for optimal performance, ileal nutrient digestibility and bone mineralization from 1 to 21 days of age. A total of 750 one-day-old male broiler chickens (Ross 308) were randomly distributed into 50 cages including 9 treatments, each replicated 5 times (except central treatment with 10 replicates) and 15 birds in each cage by CCD. The dietary Ca levels of 4.3, 5.6, 8.6, 11.7 and 13.0 g/kg and NPP of 2.5, 2.9, 4.0, 5.0 and 5.4 g/kg were used for nine treatments of CCD. The results indicated that the linear and quadratic effects of NPP, quadratic effects of Ca and Ca × NPP were significant for average weight gain (AWG, P < 0.05), average feed intake (AFI, P < 0.05), feed conversion ratio (FCR, P < 0.05) and Ca and P apparent ileal digestibility (AID, P < 0.05); however, the linear effect of Ca was significant only for FCR (P < 0.05). On the other hand, tibia and toe ash were affected by NPP (linear and quadratic, P < 0.01) and Ca (quadratic, P < 0.01). The second-order polynomial regression model was significant for AWG (R² = 0.93, P < 0.001), AFI (R² = 0.88, P < 0.001), FCR (R² = 0.78, P < 0.001), AID of Ca (R² = 0.78, P < 0.001) and P (R² = 0.88, P < 0.001), tibia ash (R² = 0.86, P < 0.001) and toe ash (R² = 0.85, P < 0.001). The multi-objective optimization indicated that broiler chickens from 1 to 21 days of age need 7.03 and 4.47 g/kg of Ca and NPP, respectively, to achieve optimal AWG, FCR, tibia and toe ash. However, the dietary Ca and NPP levels can be reduced to 6.57 and 3.95 g/kg with a slight negative impact on performance and bone mineralization, respectively. In conclusion, the findings indicate that using multi-objective optimization model such as RSM provides more information regarding optimum Ca and NPP requirements of broiler chickens, considering the complex interaction between these two minerals. While the NPP levels are in line with current recommended requirements, Ca levels are considerably lower and suggest that current recommended Ca requirements may be in excess of the needs of the broiler.     

Effects of feed access after hatch and inclusion of fish oil and medium chain fatty acids in a pre-starter diet on broiler chicken growth performance and humoral immunity

Delayed feed and water access is known to impair growth performance of day old broiler chickens. Although effects of feed access on growth performance and immune function of broilers have been examined before, effects of dietary composition and its potential interaction with feed access are hardly investigated. This experiment aimed to determine whether moment of first feed and water access after hatch and pre-starter composition (0 to 7 days) affect growth rate and humoral immune function in broiler chickens. Direct fed chickens received feed and water directly after placement in the grow-out facility, whilst delayed fed chickens only after 48 h. Direct and delayed fed chickens received a control pre-starter diet, or a diet containing medium chain fatty acids (MCFA) or fish oil. At 21 days, chickens were immunized by injection of sheep red blood cells. The mortality rate depended on an interaction between feed access and pre-starter composition (P=0.014). Chickens with direct feed access fed the control pre-starter diet had a higher risk for mortality than chickens with delayed feed access fed the control pre-starter diet (16.4% v. 4.2%) whereas the other treatment groups were in-between. BW gain and feed intake till 25 days in direct fed chickens were higher compared with delayed fed chickens, whilst gain to feed ratio was lower. Within the direct fed chickens, the control pre-starter diet resulted in the highest BW at 28 days and the MCFA pre-starter diet the lowest (Δ=2.4%), whereas this was opposite for delayed fed chickens (Δ=3.0%; P=0.033). Provision of MCFA resulted in a 4.6% higher BW gain and a higher gain to feed ratio compared with other pre-starter diets, but only during the period it was provided (2 to 7 days). Minor treatment effects were found for humoral immune response by measuring immunoglobulins, agglutination titers, interferon gamma (IFN-γ), and complement activity. Concluding, current inclusion levels of fish oil (5 g/kg) and MCFA (30 g/kg) in the pre-starter diet appear to have limited (carryover) effects on growth and development, as well as on humoral immune function.     

The effect of alternative feeding strategies on the feeding motivation of broiler breeder pullets

Due to genetic selection for fast growth and high breast meat yield, commercial strains of broiler chickens and broiler breeders are predisposed to high feed intake; however, feeding broiler breeders ad libitum impairs their health and reproductive performance. Broiler breeders are feed-restricted throughout rearing to maintain health and performance, yet feed restriction results in hunger, feeding frustration and lack of satiety. The objective of this study was to examine the effect of alternative feeding strategies, including feed additives (separately or combined) and a fixed non-daily feeding schedule, on the feeding motivation and welfare of broiler breeders during rearing. At 3 weeks of age, 180 Ross 308 breeder pullets were allocated to 90 cages and fed with one of five isocaloric treatments: (1) daily control diet (control), (2) daily calcium propionate diet (CaP), (3) daily soybean hull diet (SBH), (4) daily alternative diet (alternative: CaP + SBH) and (5) 4/3 control diet (four on-feed days and three non-consecutive off-feed days per week). The CaP diet included calcium propionate at 1.4% from 3 to 6 weeks of age, and at 3.2% from 7 to 12 weeks of age, and the SBH diet contained soybean hulls included at 40%. The alternative diet included both soybean hulls and calcium propionate at the same inclusion rate as the SBH and CaP diets, respectively. Pullets were weighed and scored for feather coverage every week. A feed intake test was conducted at 3, 4, 8, 10 and 11 weeks of age for 10 min during on- and off-feed days. At 12 weeks of age, feather samples were analysed for fault bars. Data were analysed using linear mixed regression models, with cage nested in the models and age as a repeated measure. At 4 weeks of age, pullets fed soybean hull-enriched diets (SBH and alternative diets) and those on the 4/3 schedule had lower feed intake than control pullets (P = 0.02). Feathers from pullets fed the SBH diet had fewer fault bars than those fed the CaP diet (P = 0.04). The results indicated that the inclusion of soybean hulls (alone or combined with calcium propionate) and a 4/3 feeding schedule can reduce feeding motivation of broiler breeders during early rearing.     

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.     

First report of the apparent metabolisable energy value of black soldier fly larvae fat used in broiler chicken diets

In the available literature, there are limited data about the energetic value of insect-derived products. In particular, insect fat cannot be used in practical broiler nutrition due to the lack of precise apparent metabolisable energy (AME) value. Therefore, the present study aimed to investigate the AME and apparent metabolisable energy corrected to zero nitrogen balance (AME_N) levels of Hermetia illucens larvae fat for broiler chickens of various ages. A total of 400 1-day-old male Ross 308 chicks were randomly allotted to four dietary groups (10 replicate pens per treatment; 10 birds per pen). The following treatments were applied: HI0 – basal diet without dietary fat inclusion, HI03 – basal diet enriched with 30 g/kg H. illucens larvae fat, HI06 – basal diet enriched with 60 g/kg H. illucens larvae fat, and HI09 – basal diet enriched with 90 g/kg H. illucens larvae fat. Broilers had ad libitum access to mash form feed and water. Excreta samples were collected on d 14, d 28, and d 35. To determine the AME and AME_N values of H. illucens larvae fat, the simple linear regression method was used. The results show that the AME and AME_N values of H. illucens larvae fat for broiler chickens are 9 049 kcal/kg (37.86 MJ/kg) and 9 019 kcal/kg (37.74 MJ/kg), respectively. Additionally, because the birds’ age significantly (P < 0.001) affected the AME and AME_N levels, the implementation of H. illucens larvae fat to broiler diets should be considered in each nutritional period using the recommended regression model AME = 2 559.758 + 62.989 × fat inclusion (%) + 7.405 × day of age and AME_N = 2 543.2663 + 62.8649 × fat inclusion (%) + 7.3777 × day of age. The present data emphasised that the H. illucens larvae fat metabolisable energy is similar to that of soybean oil.     

The use of an optimised concentration of quercetin limits peroxidation of lipids in the meat of broiler chickens fed a diet containing flaxseed oil rich in omega-3

Ross 308 chickens were used to investigate fatty acid (FA) composition and oxidative stability of broiler chicken meat following the controlled inhibition of peroxidation in feed containing a concentrated source of omega-3 fatty acids (flaxseed oil, FLO), approximately 50% omega-3. Ninety-six one-day-old chicks were randomly allocated to four dietary treatments (n = 24/group) that included rapeseed oil (RO), flaxseed oil (FLO), RO with optimised quercetin (Q) – RO_Q, or FLO with optimised Q (FLO_Q). On day 35, breast and thigh muscles were collected in order to analyse their FA profile and malondialdehyde (MDA) levels. Dietary treatments had no effect on weight gain or feed conversion ratio in chickens. However, dietary FLO increased the deposition of α-linolenic acid in both pectoral and thigh meat (P < 0.05), and tended to decrease the ratio of omega-6/omega-3 FA in pectoral muscles (P < 0.07). Addition of an optimised concentration of Q proved to be an efficient way of limiting lipoperoxidation in breast and thigh muscles subjected to refrigeration at 2–3 °C for either 1 or 7 days. Results were consistent with the observed inhibition of peroxidation in feed mixtures and significantly correlated with MDA levels found in feed mixtures. These results provide evidence that it is possible to produce poultry meat with an improved proportion of omega-3 FA without significantly altering the performance of broiler chickens or the oxidative stability of their meat.     

Effect of chronic heat stress on gastrointestinal histology and expression of feed intake-regulatory hormones in broiler chickens

Heat stress (HS) dramatically impairs the growth performance of broiler chickens, mainly as a consequence of reduced feed intake due to the loss of appetite. This study was aimed at evaluating the alterations induced by chronic HS conditions on the morphological and morphometric features of the gastrointestinal (GI) tract and on the expression of some enteroendocrine cells (EECs) involved in the regulation of feed intake in chickens. Three hundred male chickens (Ross 308) were divided into two experimental groups and raised either in thermoneutral environment for the whole fattening period (0–41 days) (TNT group) or subjected to chronic HS conditions (30 °C for 24 h/day) from 35 to 41 days (HS group). Samples of proventriculus, duodenum, jejunum and cecum were collected from 24 broilers (12/group). Haematoxylin-eosin was used for the morphometric evaluations, while immunohistochemistry was applied for the evaluation of EECs expressing ghrelin (GHR), cholecystokinin (CCK), neuropeptide Y (NPY), glucagon-like peptide-1 (GLP-1), and serotonin (5-HT). In the proventriculus, HS reduced total wall thickness and mucous layer height (P ≤ 0.01) as well as mean diameter, circumference, and area of the compound tubular glands (P ≤ 0.001) with respect to TNT. The small intestine of HS birds was characterised by decreased villous height and total thickness (duodenum, P ≤ 0.01; jejunum, P ≤ 0.001), whereas crypt depth and width were reduced only in the jejunum (P ≤ 0.01). HS had negligible effects on the morphological aspects of the cecum. In the proventriculus, an increase in GHR and NPY EECs was observed in response to HS (P ≤ 0.001). Similarly, the small intestine villi of the HS group showed greater GLP-1 (P ≤ 0.05), 5-HT (P ≤ 0.001) and CCK (P ≤ 0.01) EECs. Moreover, the expression of 5-HT EECs was higher in the duodenal (P ≤ 0.01) and jejunal (P ≤ 0.01) crypts of HS birds, whereas GLP-1 and CCK EECs increased only in jejunal crypts (P ≤ 0.05). Finally, 5-HT EEC expression was increased in the cecum of HS group (P ≤ 0.01). In conclusion, these outcomes demonstrate that chronic HS induces morphometric alterations not only in the small intestine but also in a key organ such as the proventriculus. Furthermore, HS conditions affect the presence and distribution of EECs, suggesting that some GI peptides and biogenic amine may be implicated in the regulation of appetite and voluntary feed intake in heat-stressed broiler chickens.     

Nanoencapsulation (in vitro and in vivo) as an efficient technology to boost the potential of garlic essential oil as alternatives for antibiotics in broiler nutrition

The addition of essential oil (EO) as chitosan encapsulated can increase the efficiency of these oils in broiler feeding. Therefore, the objective of the current research was to explore the antibacterial and antioxidant potential of garlic essential oil (GEO) (free vs. nanoencapsulated) and their effects on performance, gene expression of mucin2, microbial, and morphology of intestine in broilers. A total of 900 1-day-old male broilers (Ross 308) were assigned to six dietary treatments (0, 100, and 200 mg/kg free GEO and 0 (contain of chitosan), 100 and 200 mg/kg nanoencapsulated GEO) with a 2 × 3 factorial arrangement based on completely randomized design. Garlic essential oil encapsulation with chitosan significantly enhanced antibacterial and antioxidant parameters. At 100 mg/kg nanoencapsulated GEO had significant (P < 0.01) advantages in improving BW gain (BWG) (22–42 and 0–42) and feed conversion ratio (FCR) (0–42). Maximum feed intake (FI) was also associated with the control group (P < 0.05). Broilers fed on 100 mg/kg of nanoencapsulated GEO showed higher villi length and width relative to other treatments and villi length to crypt depth ratio as well (P < 0.01). The nanoencapsulation process of GEO (P < 0.01) affected the Lactobacilli population in the digesta of ileo-caecum and mucin2 gene expression. In broiler chickens, the tested EO, especially nanoencapsulated type, enhanced more evaluated parameters. Because of its ideal properties, nanoencasulating with chitosan may also be an effective and inexpensive way to protect bioactive compounds and improve GEO effects in broiler chickens.     

The effect of Candida famata and Lactobacillus plantarum on the number of coliforms and the antibiotic resistance and virulence of Escherichia coli in the gut of broilers

This study was undertaken to determine the effect of a yeast (Candida famata) and a bacterium (Lactobacillus plantarum), administered alone or in combination in the drinking water, on the population of yeast, Lactobacillus sp. and coliforms, and the prevalence of antimicrobial resistance (AMR) and virulence genes in Escherichia coli (E. coli) isolated from digesta samples taken throughout the life of broiler chickens. Male (Ross 308) day-old chicks (220) were used. C. famata (isolated from a chicken) and L. plantarum (isolated from a pig) were administered via the drinking water. Water was provided either untreated or with C. famata (CF; 10⁸/ml), L. plantarum (LP; 10⁵–10⁸/ml), or a combination of CF and LP (10⁶–10⁸/ml) in water hoppers on 2 days each week for 35 days. Administering probiotics did not affect the growth performance in broiler chickens. No significant interactions were observed between main effects, and neither CF nor LP had any effect on the population size of Lactobacillus sp. or coliforms. The administration of C. famata increased the population density of yeasts in the small intestine at these ages. The population density of coliforms, Lactobacillus sp. and yeast decreased with age (P < 0.001). There was no significant effect of probiotics on the prevalence of phenotypic AMR and virulence genes in these studies. The prevalence of E. coli that was resistant to ampicillin and tetracycline, as well as carrying ≥3 virulence-associated genes, was greatest at the end of the starter phase (around 8 days old), before declining through the grower and finisher phases. There was only limited evidence that administering either CF or LP affected either the AMR or the virulence of E. coli in the bird. However, tetracycline resistance in E. coli was associated (P < 0.001, P < 0.01, P < 0.05, and P < 0.05) with the carriage of the iron uptake systems of E. coli D, iron-repressible protein, increased serum survival and temperature-sensitive haemagglutinin genes respectively, suggesting that the accumulation of iron and the genetic element conferring tetracycline resistance may be intertwined.     

Early age thermal manipulation on the performance and physiological response of broiler chickens under hot humid tropical climate

Initial brooding temperature is critical for post-hatch growth of broiler chickens. A study was conducted to investigate the early age thermal manipulation (EATM) on the performance and physiological responses broiler chickens under hot humid tropical climate. A total of 260 unsexed day-old Arbor-acre broiler chicks were assigned to five thermal treatments of brooding temperature regimens having 4 replicates of thirteen birds each. The heat treatments were: initial brooding temperature of 35 °C for the first 2 days, and then decreased subsequently, gradually to 22 °C at 21 d of age (CT), initial temperature of 35 °C, sustained for the first 4 days and then decreased gradually (conventionally) (FD), initial temperature of 35 °C for the first 7 days (SD), the birds in CT, but the brooding temperature was raised to 35 °C again for another 3 days from day 7 (SD3), initial brooding temperature of 35 °C for the first 10 days (TD). Data were collected on daily feed intake and weekly body weights. Blood samples were collected from 8 birds per treatment weekly for the determination of plasma uric acid, triglycerides, triiodothyronine (T₃) and creatinine kinase. Data obtained were laid out in a Completely Randomized Design (CRD). Results showed that the final weights of the birds in FD were higher (P < 0.05) than those of the other treatments at the finisher phase. Feed intake of the birds in FD was higher than those of SD3 and TD. FCR of broiler chickens in CT, SD, SD3 and TD was higher than that of FD. The rectal temperature, plasma MDA and blood glucose of the thermally challenged birds in FD was generally better (P < 0.05) than those of the other treatments. It was concluded that EATM can be used to improve performance and also protect broiler chickens from acute heat stress at market age.     

The effect of level of straw bale provision on the behaviour and leg health of commercial broiler chickens

This study aimed to assess the effect of the number of straw bales (SBs) provided on the behaviour and leg health of commercial broiler chickens. Houses containing ~23 000 broiler chickens were assigned to one of two treatments: (1) access to 30 SBs per house, ‘30SB’ or (2) access to 45 SB per house, ‘45SB’. This equated to bale densities of 1 bale/44 m² and 1 bale/29 m² of floor space within houses, respectively. Treatments were applied in one of two houses on a commercial farm, and were replicated over six production cycles. Both houses had windows and were also artificially lit. Behaviour was observed in weeks 3 to5 of the cycle. This involved observations of general behaviour and activity, gait scores (0: perfect to 5: unable to walk) and latency to lie (measured in seconds from when a bird had been encouraged to stand). Production performance and environmental parameters were also measured. SB density had no significant effect on activity levels (P>0.05) or walking ability (P>0.05). However, the average latency to lie was greater in 30SB birds compared with 45SB birds (P<0.05). The incidence of hock burn and podo dermatitis, average BW at slaughter and levels of mortality and culling were unaffected by SB density (P>0.05). The results from this study suggest that increasing SB levels from 1 bale/44 m² to 1 bale/29 m² floor space does not lead to significant improvements in the welfare of commercial broiler chickens in windowed houses.     

Influence of level of inclusion of Azolla leaf meal on growth performance,meat quality and skeletal muscle p70S6 kinase α abundance in broiler chickens

The interest in biodiesel production from oil-bearing seeds rather than soybean necessitates the scientific validation of other good quality protein sources that could substitute soybean meal in animal diets, particularly, broiler chickens where soybean meal constitutes a large portion of their diet. Therefore, the present study was conducted to investigate the effect of sun-dried Azolla leaf meal (ALM) as an unconventional dietary protein source in broiler chicken diet on growth performance, meat quality, skeletal muscle cell growth and protein synthesis through regulation of ribosomal protein S6 kinase (p70S6 kinase α). A total of 120 male Ross 308 broiler chicks were randomly allocated to three dietary treatments. Each treatment had four cages (i.e. replicates) with 10 birds/cage. The control group was fed with a corn–soy-based diet, the AZ5 group was supplemented with 5% ALM and the AZ10 group was supplemented with 10% ALM for 37 days. A 5-day trial was also conducted to measure the apparent nutrient digestibility. Growth performance parameters were measured weekly. At the end of the experiment, 12 birds from each group (3/cage) were euthanized and used for samplings. Inclusion of ALM tended to improve BW gain (P = 0.06) and increased feed intake (P < 0.01). Additionally, ALM decreased the percentage of breast meat cooking loss linearly (P < 0.01). In addition, ALM at a dose of 5% increased the production of propionate in the cecum (P = 0.01). Activation of breast muscle p70S6 kinase was higher when ALM was included in a dose-dependent manner (P < 0.01). The inclusion of ALM increased breast meat redness (P < 0.01); however, the lightness was within the normal range in all groups. Findings from our study suggest that ALM could be included in a broiler chicken diet up to 5% without any major negative effect on meat quality or performance, and it regulates muscle protein synthesis through activation of mammalian target of rapamycin/6S kinase signaling.     

Associations between behaviour and health outcomes in conventional and slow-growing breeds of broiler chicken

Broiler chickens are prone to a range of complex health and welfare issues. To support informed selection of welfare traits whilst minimising impact on production efficiency and to address a major gap in understanding, we systematically explored associations between health and behavioural indicators of broiler welfare. One conventional (CNV, n = 350) and two slow-growing broiler breeds (SGH and SGN, respectively n = 400) were reared from hatch in pens of 50 birds. Birds were assessed for health (gait, plumage cover and dirtiness, pododermatitis, hockburn, and leg deviations) at 2.2 kg liveweight according to the Royal Society for the Prevention of Cruelty to Animals Broiler Breed Welfare Assessment Protocol. Behaviour and resource-use of 10% of birds per pen, on days 29 (all breeds) and 43 (SGH and SGN), was (i) scan sampled every 60 min between three to six and between twelve to fifteen hours after photoperiod onset; and (ii) continuously sampled sequentially from focal birds for 3 min each in a random order, during 15 min observation periods at three and twelve hours after photoperiod onset. Binary logistic generalized linear models were used, to assess respective associations between pen prevalence of each health outcome and (i) pen mean percentage scans of behaviour, and (ii) pen mean frequency and duration per 3 min focal observation of behaviour. Better growth rate and feed conversion but poorer health outcomes (mortality, gait, pododermatitis, feather cover) were more prevalent in CNV. Strong associations between behaviour and several heath indicators revealed, (i) increases in side-lying inactive, sitting inactive, and use of the litter relative to other resources, as primary and general indicators of poorer health, and (ii) increases in standing inactive, perch use, walking, Comfort, High Energy and Exploratory behaviour as primary and general indicators of better health. Of these, changes in side-lying, standing inactive, walking, Comfort and High Energy behaviour were particularly sensitive to small differences in health outcomes important for breed acceptance in high-welfare schemes. Crucially these behavioural measures additionally represent motivational and affective aspects of welfare not captured by health measures and allow opportunity for earlier intervention. Thus, to provide a comprehensive assessment of broiler experience, behaviour should be incorporated into broiler welfare assessments.     

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.     

Early enrichment in free-range laying hens: effects on ranging behaviour,welfare and response to stressors

Free-range laying hen systems are increasing within Australia. The pullets for these systems are typically reared indoors before being provided first range access around 21 to 26 weeks of age. Thus, the rearing and laying environments are disparate and hens may not adapt well to free-range housing. In this study, we reared 290 Hy-Line® Brown day-old chicks divided into two rooms each with feed, water and litter. In the enriched room, multiple structural, manipulable, visual and auditory stimuli were also provided from 4 to 21 days, the non-enriched room had no additional objects or stimuli. Pullets were transferred to the laying facility at 12 weeks of age and divided into six pens (three enriched-reared, three non-enriched-reared) with identical indoor resources and outdoor range area. All birds were first provided range access at 21 weeks of age. Video observations of natural disturbance behaviours on the range at 22 to 23 and 33 to 34 weeks of age showed no differences in frequency of disturbance occurrences between treatment groups (P=0.09) but a decrease in disturbance occurrences over time (P<0.0001). Radio-frequency identification tracking of individually tagged birds from 21 to 37 weeks of age showed enriched birds on average, spent less time on the range each day (P<0.04) but with a higher number of range visits than non-enriched birds from 21 to 24 weeks of age (P=0.01). Enriched birds accessed the range on more days (P=0.03) but over time, most birds in both treatment groups accessed the range daily. Basic external health scoring showed minimal differences between treatment groups with most birds in visibly good condition. At 38 weeks of age all birds were locked inside for 2 days and from 40 to 42 weeks of age the outdoor range was reduced to 20% of its original size to simulate stressful events. The eggs from non-enriched birds had higher corticosterone concentrations following lock-in and 2 weeks following range reduction compared with the concentrations within eggs from enriched birds (P<0.0001). Correspondingly, the enriched hens showing a greater increase in the number of visits following range area reduction compared to non-enriched hens (P=0.02). Only one rearing room per treatment was used but these preliminary data indicate 3 weeks of early enrichment had some long-term effects on hen ranging behaviour and enhanced hen’s adaptability to environmental stressors.     

Enzyme supplementation of wheat-based diets for broilers: 2. Effect on apparent metabolisable energy content and nutrient digestibility

Two experiments were conducted to determine the effect of enzyme supplementation on the nitrogen-corrected apparent metabolisable energy (AME_n) and apparent nutrient digestibilities in ileum and excreta from male broiler chickens fed diets containing high amounts of wheat (>80%). Four different enzyme preparations were added to the wheat-based diets in varying levels and combinations. The difference between Experiments 1 and 2 consisted in the addition of different enzymes and enzyme levels. Excreta and ileal content were collected from broiler chickens at 3 and 6 weeks of age. At 3 weeks of age enzyme supplementation increased (P<0.0001) the AME_n from an average value of 13.86 MJ kg⁻¹ dry matter (DM) to an average of 14.60 MJ kg⁻¹ DM in the two experiments. The apparent digestibility of protein (APD) and fat (AFD) were improved significantly as a result of enzyme addition in both experiments. At 3 weeks of age, improvements (P<0.05) in the ileal AFD and APD were on average 13% and 6%, respectively. The effect of enzyme supplementation on AFD measured in excreta from broiler chickens showed the same pattern. The positive effect of enzyme addition on the overall nutrient digestibility and AME_n was reflected in weight gain and feed conversion efficiency (FCE). Significant improvements in AME_n, APD, and AFD were still present in most of the groups fed with enzyme-supplemented diets at 6 weeks of age. The effect of enzyme addition, however, was less pronounced, especially in Experiment 1 where the ileal APD of broiler chickens did not differ significantly from the control group. Apparent starch digestibility (ASD), measured in Experiment 2, was very high in all groups, including the control; however, enzyme supplementation increased ASD (P<0.001) in ileum and excreta at both 3 and 6 weeks of age. The digestibility of total non-starch polysaccharides (NSP) in excreta was improved significantly (P<0.01) as a result of enzyme supplementation (Experiment 2). In addition, pH of caeca content decreased (P<0.02) in broiler chickens fed with enzyme-supplemented diets when compared with the control group. Decreased pH could indicate microbial fermentation of unabsorbed NSP residues and nitrogenous compounds. Overall, the results demonstrated that the nutritive value of wheat-based diets to broiler chickens improve enzyme supplementation. The apparent digestibility of total NSP and of arabinose and xylose residues improved (P<0.02) in the enzyme-supplemented diets indicating that the enzymes were able to break down the cell wall NSP to a certain extent.     

Breast muscle and plasma metabolomics profile of broiler chickens exposed to chronic heat stress conditions

Understanding the variations of muscle and plasma metabolites in response to high environmental temperature can provide important information on the molecular mechanisms related to body energy homeostasis in heat-stressed broiler chickens. In this study, we investigated the effect of chronic heat stress conditions on the breast muscle (Pectoralis major) and plasma metabolomics profile of broiler chickens by means of an innovative, high-throughput analytical approach such as the proton nuclear magnetic resonance (¹H NMR) spectrometry. A total of 300 Ross 308 male chicks were split into two experimental groups and raised in either thermoneutral conditions for the entire rearing cycle (0–41 days) (TNT group; six replicates of 25 birds/each) or exposed to chronic heat stress conditions (30 °C for 24 h/day) from 35 to 41 days (CHS group; six replicates of 25 birds/each). At processing (41 days), plasma and breast muscle samples were obtained from 12 birds/experimental group and then subjected to ¹H NMR analysis. The reduction of BW and feed intake as well as the increase in rectal temperature and heterophil: lymphocyte ratio confirmed that our experimental model was able to stimulate a thermal stress response without significantly affecting mortality. The ¹H NMR analysis revealed that a total of 26 and 19 molecules, mostly related to energy and protein metabolism as well as antioxidant response, showed significantly different concentrations respectively in the breast muscle and plasma in response to the thermal challenge. In conclusion, the results obtained in this study indicated that chronic heat stress significantly modulates the breast muscle and plasma metabolome in fast-growing broiler chickens, allowing to delineate potential metabolic changes that can have important implications in terms of body energy homeostasis, growth performance and product quality.     

Effects of Corticosterone and Dietary Energy on Immune Function of Broiler Chickens

An experiment was conducted to investigate the effects of dietary energy level on the performance and immune function of stressed broiler chickens (Gallus gallus domesticus). A total of 96 three-day-old male broiler chickens (Ross × Ross) were divided into two groups. One group received a high energy (HE) diet and the other group received a low energy (LE) diet for 7 days. At 5 days of age, the chickens from each group were further divided into two sub-groups and received one of the following two treatments for 3 days: (1) subcutaneous injection of corticosterone, twice per day (CORT group; 2 mg of CORT/kg BW in corn oil) and (2) subcutaneous injection of corn oil, twice per day (Control/Sham treatment group). At 10 days of age, samples of blood, duodenum, jejunum, and ileum were obtained. Compared with the other three groups, the LE group treated with CORT had the lowest average daily gain (ADG) and the poorest feed conversion ratio (FCR, P < 0.05). Furthermore, CORT treatment decreased the relative weight (RW) of the bursa independent of the dietary energy level, but it decreased the RW of the thymus only in the chickens fed the LE diet. By contrast, CORT administration decreased the RW of the spleen only in the chickens fed the HE diet (P < 0.05). The plasma total protein, albumin, tumor necrosis factor alpha, interleukin 2 and immunoglobulin G (IgG) levels were affected by the CORT treatment (P < 0.05); however, these factors were not significantly affected by the dietary energy level. Toll-like receptor-5 mRNA level was down-regulated by CORT injection in the duodenum and ileum (P < 0.05) and showed a trend of down-regulation in the jejunum (P=0.0846). The present study showed that CORT treatment induced immunosuppressive effects on the innate immune system of broiler chickens, which were ameliorated by consumption of higher dietary energy.     

Effect of Campylobacter-Specific Maternal Antibodies on Campylobacter jejuni Colonization in Young Chickens

Using laboratory challenge experiments, we examined whether Campylobacter-specific maternal antibody (MAB) plays a protective role in young chickens, which are usually free of Campylobacter under natural production conditions. Kinetics of C. jejuni colonization were compared by infecting 3-day-old broiler chicks, which were naturally positive for Campylobacter-specific MAB, and 21-day-old broilers, which were negative for Campylobacter-specific MAB. The onset of colonization occurred much sooner in birds challenged at the age of 21 days than it did in the birds inoculated at 3 days of age, which suggested a possible involvement of specific MAB in the delay of colonization. To further examine this possibility, specific-pathogen-free layer chickens were raised under laboratory conditions with or without Campylobacter infection, and their 3-day-old progenies with (MAB⁺) or without (MAB⁻) Campylobacter-specific MAB were orally challenged with C. jejuni. Significant decreases in the percentage of colonized chickens were observed in the MAB⁺ group during the first week compared with the MAB⁻ group. These results indicate that Campylobacter-specific MAB plays a partial role in protecting young chickens against colonization by C. jejuni. Presence of MAB in young chickens did not seem to affect the development of systemic immune response following infection with C. jejuni. However, active immune responses to Campylobacter occurred earlier and more strongly in birds infected at 21 days of age than those infected at 3 days of age. Clearance of Campylobacter infection was also observed in chickens infected at 21 days of age. Taken together, these findings (i) indicate that anti-Campylobacter MAB contributes to the lack of Campylobacter infection in young broiler chickens in natural environments and (ii) provide further evidence supporting the feasibility of development of immunization-based approaches for control of Campylobacter infection in poultry.     

Effects of a riboflavin source suitable for use in organic broiler diets on performance traits and health indicators

Riboflavin (vitamin B₂) is essential for monogastric animals. It is mainly produced by recombinant microorganisms (Candida famata, Bacillus subtilis and Ashbya gossypii). The availability of genetically modified organism (GMO)-free riboflavin, obligatory in European organic agriculture, is a major issue. Besides, requirements for organic livestock might differ from conventional production because other genotypes and feed formulations are used. The effects of a fermentation suspension with a high native content of riboflavin produced with unmodified A. gossypii by fermentation were investigated at graded dosages as an alternative to conventional (GMO-based) riboflavin in slow-growing broilers on performance traits and health and welfare indicators. In 2 runs with 800 animals each, Ranger Gold™ broilers were fed with 4 dietary treatments. For starter diets (day 1 to 18), treatments included a basal diet (1) without any riboflavin supplementation (negative control, N-C), (2) with conventional riboflavin supplementation (Cuxavit B2 80% riboflavin) at 9.6 mg/kg (positive control, P-C), (3) with riboflavin supplementation from the alternative source at 3.5 mg/kg (A-low) and (4) with riboflavin supplementation from the alternative source at 9.6 mg/kg (A-high). For the finisher diet (day 29 until slaughtering), P-C and A-high were supplemented with 8.0 mg/kg and A-low with 3.5 mg/kg. Diets were formulated according to organic regulations. Animals were kept in floor pens with 20 chickens per pen. Weekly, BW, feed and water consumption were recorded. Every second week, animal-based health and welfare indicators (feather score and footpad dermatitis) were scored. Slaughter traits were assessed for five males and females per pen at 62/63 days of age. Final body weight of A-high differed from N-C and A-low, but not from P-C. From week 2 until six years of age, A-high had a higher daily weight gain when compared to all other groups. With 74.4%, dressing percentage was higher in A-high compared with all other groups (73.3%). Breast percentage of A-low was lower than that of both control groups but did not differ from A-high. The highest frequency of liver scores indicating fatty liver syndrome was found in P-C, followed by N-C and A-low. Feather scores did not respond to treatment; the highest frequency of mild footpad dermatitis was observed in A-high, however at a low prevalence. In conclusion, the tested fermentation suspension with a high native content of riboflavin derived from fermentation of A. gossypii can be used at levels of commercial recommendations as alternative to riboflavin produced from GMO in broiler feeding. Further studies must verify whether riboflavin can be reduced without inducing riboflavin deficiency in slow-growing broilers.