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The Australian and global chicken meat industries have benefited from rapid improvements in the efficiency of chicken meat production that have been predominantly achieved through genetic selection, optimisation of bird nutrition and improved bird health. However, this has also resulted in morphological changes in the bird with an increase in the prevalence of leg health disorders. Compromised leg health can cause pain and lameness and bodes poorly for bird wellbeing, bird mortality, and economic returns. There are also implications for the consumer who is increasingly mindful of animal welfare and is demanding more welfare friendly products. Accurate on-farm assessment of bird leg health has challenges due to the diversity of leg disorders and the variety of techniques used to assess their severity and impact. Overall prevalence of leg disorders shows great variability between properties (farms) and flocks. Opportunities to improve bird leg health have been the focus of considerable research which has frequently included an evaluation of environmental enrichment as a means to reduce lameness and improve bird mobility. To this end, currently in Australia, 78% of chicken meat is produced under the conditions of the Australian RSPCA Approved Farming Scheme, which requires perches in the birds’ environment. However, the value of perches in providing enrichment and improving bird welfare is unclear. Therefore, this review explores animal welfare and consumer attitudes towards meat chicken welfare, describes leg disorders, outlines techniques for assessing leg health and discusses opportunities to enrich the birds’ environment to improve bird mobility and leg health.

Previous work has identified an effect of hatch time on chick femur mineralization. This experiment assessed the impact of hatch time and a 24-h post-hatch unfed time period on chick bone mineralization and yolk mineral utilization. In early hatching chicks, yolk Mg, Zn, K, P, Fe, and Cu decreased by 40 to 50% over the 24-h post-hatch unfed time period, whereas yolk Ca and Na decreased by 25 to 40% (P = 0.026). Yolk Sr was intermediate, decreasing by 37%. Late hatching chicks which had been hatched for no more than 30 h had a higher femur bone ash percentage compared to early hatching chicks which had spent over a 30-hour sojourn unfed in the incubator (P = 0.013). These results indicate that removing chicks from the incubator within 30 h of their hatch is likely to benefit their femoral mineralization.

Lower egg shell temperatures (EST) during the first 2 weeks of incubation, notionally known as Slow start incubation, extended the standing time of a 5-week-old fast feathering meat chicken parent line. This study was designed to evaluate the effect of Slow start incubation on the standing ability of commercial meat chickens. Eggs from two strains of meat chickens, Strains 1 and 2, were incubated using either the Slow start incubation, (the initial EST was 36.75°C followed by a gradual increase in EST, reaching 37.8°C at day 16 of incubation), or Control incubation (EST 37.75°C to 38°C from the start of incubation until day 18 of incubation). Eggs were observed every 6 h from 468 h until 516 h of incubation to identify chick hatch window. At 516 h of incubation all chicks were taken out of the incubator (take-off). Chicks from each Strain and incubation treatment were randomly selected for assessment of chick weight, chick length, yolk sac weight, serum Ca and P, and femoral bone ash (BA). All unhatched eggs were inspected to determine the stage of embryo failure. Remaining chicks were grown for 5 weeks in floorpens. Weekly feed intake (FI), chick weight and feed conversion ratio (FCR) were determined. At 35 days of age the standing ability of visibly male birds was assessed in a latency-to-lie test. Compared to the Control, Slow start incubation delayed the average hatch time of both strains by ∼13 h, and reduced hatchability with 4.6% live but unhatched chicks, which was most evident in Strain 2. Significant differences due to main effects only were observed at take-off. Strain 1 chicks were significantly heavier and longer with higher serum Ca but significantly lower BA and serum P than Strain 2. Slow start incubation generated significantly heavier chicks that were shorter, but had significantly heavier yolk sacs, lower serum Ca but higher serum P than Control incubated chicks. During the 1st week post hatch Strain 1 Control incubated chicks had significantly higher FI and higher FCR than all other Strain and incubation treatments. At 35 days of age Slow start incubated birds of both Strains stood significantly longer than those from the Control incubation. This experiment clearly demonstrated the ability of Slow start incubation of commercial meat chickens to improve their leg strength.

A total of 864 settable Cobb 500 eggs were used to explore changes in yolk mineral content during incubation. Eggs were individually weighed and then placed in a commercial incubator. On embryonic day (ED) 0, 6.5, 13.5, and 17.5, 36 eggs were sampled and yolk weight and mineral content were determined. The concentration of iron (Fe), phosphorus (P), and zinc (Zn) declined (P < 0.05) from ED0 to ED17.5. The concentration of calcium (Ca), magnesium (Mg), and strontium (Sr) increased (P < 0.05) from ED0 to ED17.5. The concentration of copper (Cu), potassium (K), and sodium (Na) increased initially (ED0 to ED6.5) but declined thereafter. There was no change (P > 0.05) in the concentration of yolk manganese (Mn) from ED0 to ED17.5. Substantial changes in yolk mineral concentration occur during incubation and are presumably associated with mobilization of shell reserves and flux between albumen and yolk. These data may be useful in designing in ovo interventions, optimizing meat chicken breeder premix formulation or assembly of suitable neonatal or pre-starter diets for meat chicken chicks.

Slow-growing broilers offer differentiation in the chicken meat market for consumers who have distinct preferences based on perceived higher welfare indices and willingness to pay a higher price for the product. Although breeding for slow-growing broilers is relatively advanced in Europe and the United States, it is limited in Australia. Crossbreeding is one of the approaches taken to developing slow-growing broiler strains. Thus, the aim of this study was to compare performance, immune response, leg health, carcass characteristics, and meat quality of a novel crossbred slow-growing broiler breed (SGB) with the conventional, fast-growing Cobb 500 broiler (CB) to assess their suitability as an alternative for chicken meat production in Australia. A total of 236 one-day-old broiler chicks (116 SGB and 120 fast-growing CB) were reared on standard commercial diet in an intensive production system. Birds and feed were weighed on a weekly basis and feed intake and feed conversion ratio calculated. At 21 d of age, a 2% suspension of sheep red blood cells was injected subcutaneously into 8 broilers of each breed to compare their antibody response. Birds from both breeds were grown to a final live weight of 2.0–2.2 kg, before a latency-to-lie (LTL) test, carcass analysis and apparent metabolizable energy (AME) assay were performed. The SGB reached the target weight at 55 d of age compared with 32 d in CB. However, SGB stood for longer during LTL, had higher thigh, drumstick, and wing yields (as a percentage of carcass weight) as well as darker and redder meat in comparison with the CB. The CB had better feed conversion efficiency, higher antibody (IgM) production, higher AME, heavier breast yield, and lower meat drip loss than the SGB. Although fast-growing CB outperformed the SGB for traditional performance parameters, the crossbred in this study was comparable with other slow-growing broiler breeds and strains across different countries and is thus a suitable candidate for a slow-growing alternative in Australia.

Fertile eggs from Cobb 500 broiler breeder hens were incubated to provide low starting egg shell temperatures (EST; 36.9°C to 37.3°C) which were gradually increased to 37.8°C during the first 7 to 15 days of incubation compared with eggs incubated with a constant EST of 37.8°C (standard conditions) over the first 18 days of incubation. Time of individual chick hatching (measured at 6 h intervals from 468 h of incubation), chick weight, chick length and yolk weight were measured at take-off and BW was measured at 7, 14, 28, 34 and 42 days of age. Male birds at 34 and 42 days of age were assessed for their ability to remain standing in a latency-to-lie test. At 34 and 42 days, male birds were examined for leg symmetry, foot pad dermatitis, hock bruising and scored (scale 0 to 4, where 0=no lesion and 4=lesions extending completely across the tibial growth plate) for tibial dyschondroplasia (TD) lesions. The lower EST profiles caused chicks to hatch later than those incubated under the standard EST profile. Chicks which hatched at ⩽498 h incubation grew faster over the first 7 days than those that hatched later. There were significantly more birds (only males were studied) that hatched from the lower EST profiles with TD scores of 0 and 1 and fewer with score 4 at 34 days than those hatched under the standard profile. Male birds at 34 days with TD lesions ⩾3 stood for significantly shorter times than males with TD scores ⩽2. Moreover, male birds at 34 and 42 days with TD lesion scores of ⩾3 hatched significantly earlier and grew significantly faster over the first 2 weeks of age than did male birds with TD scores ⩽2. It appears possible to decrease the severity and prevalence of TD in the Cobb 500 broiler by ensuring that the birds do not hatch before 498 h of incubation.

Objective In Australia, Salmonella serovar Typhimurium (S. Typhimurium) is the predominant zoonotic serovar in humans and is frequently isolated from layer hens. Vaccination against this serovar has been previously shown to be effective in broilers and the aim of this current study was to assess and determine the best vaccination strategy (live or inactivated) to minimise caecal colonisation by S. Typhimurium. Methods A long-term experiment (56 weeks) was conducted on ISABROWN pullets using a commercial live aroA deleted mutant S. Typhimurium vaccine and an autogenous inactivated multivalent Salmonella vaccine (containing serovars Typhimurium, Infantis, Montevideo and Zanzibar). These vaccines were administered PO or by SC or IM injection, either alone or in combination. Pullets were vaccinated throughout rearing (to 18 weeks of age) and sequentially bled for antibody titre levels. The birds, vaccinated and controls, were challenged orally with a field isolate of S. Typhimurium at different ages, held for 21 days post-challenge, then euthanased and their caeca cultured for the presence of Salmonella. Results None of the oral live-vaccinated groups exhibited lasting protection. When administered twice, the inactivated vaccine gave significant protection at 17 weeks of age and the live vaccine given by SC injection given twice produced significant protection at 17, 25 and 34 weeks. Conclusions Vaccination regimens that included parenteral administration of live or inactivated vaccines and thus achieved positive serum antibody levels were able to provide protection against challenge. Hence, vaccination may play a useful role in a management strategy for Salmonella carriage in layer flocks.

From ~35 days of age fast growing meat chickens spend extended periods sitting or lying and less time standing. In a fast-feathering parent line lower early incubation temperatures which delayed chick hatch time, improved bone ash and extended their standing time. This incubation study assessed the consequences of incubation temperatures, hatch time and chick management at hatch/take off on femoral bone ash (BA) in Cobb 500 meat chickens. Embryos were incubated under either Control (between 37.8°C and 38.2°C egg shell temperature (EST)) or a Slow start (from 37.2°C at sett (the start of incubation), reaching 37.8°C EST at day 13 incubation), temperatures. Hatched chicks were identified at 492 h (20.5 days of incubation – classified as early (E)) or, between >492 and ⩽516 h (>20.5 and ⩽21.5 days of incubation – classified as late (L)), from setting. The E hatch chicks were allocated across three post-hatch treatments; treatment 1: E hatch chicks that were sampled E at 492 h from setting; treatment 2: E hatch chicks that were fed for a further 24 h in a floorpen before being sampled L at 516 h from setting; treatment 3: E hatch chicks that spent a further 24 h in the incubator before being sampled L at 516 h from setting. All L hatch chicks formed one treatment group which was sampled L at 516 h (i.e. L hatch chicks sampled L). It is not possible to sample L hatching chicks E hence this treatment is absent from the experimental design. Slow start incubation resulted in a higher total hatch percentage with a greater proportion of chicks hatching L, compared with the Control incubation. The L hatching chicks had significantly higher BA than the E hatching chicks. Of the E hatching chicks, those sampled both E and L had significantly lower BA than E hatching chicks fed for 24 h before L sampling. The E hatch, fed and sampled L chicks had the numerically highest BA, which was not significantly different from the BA of the L hatching chicks sampled L These results demonstrate that BA at hatch can be improved, either by extending the incubation period through a Slow start incubation profile, inducing L hatch, or alternatively, via the prompt provision of feed to E hatching chicks.

Satellite cells (SC) are multipotential stem cells that can be induced by nutrition to alter their cellular developmental fate, which may vary depending on their fiber type origin. The objective of the current study was to determine the effect of restricting protein synthesis on inducing adipogenic transdifferentiation and apoptosis of SC originating from fibers of the fast glycolytic pectoralis major (p. major) and fast oxidative and glycolytic biceps femoris (b. femoris) muscles of the chicken. The availability of the essential sulfur amino acids Met and Cys was restricted to regulate protein synthesis during SC proliferation and differentiation. The SC were cultured and treated with 1 of 6 Met/Cys concentrations: 60/192, 30/96 (control), 7.5/24, 3/9.6, 1/3.2, or 0/0 mg/L. Reductions in Met/Cys concentrations from the control level resulted in increased lipid staining and expression of the adipogenic marker genes peroxisome proliferator-activated receptor gamma and stearoyl-CoA desaturase during differentiation in the p. major SC. Although b. femoris SC had increased lipid staining at lower Met/Cys concentrations, there was no increase in expression of either adipogenic gene. For both muscle types, SC Met/Cys, concentration above the control increased the expression of peroxisome proliferator-activated receptor gamma and stearoyl-CoA desaturase during differentiation. As Met/Cys concentration was decreased during proliferation, a dose-dependent decline in all apoptotic cells occurred except for early apoptotic cells in the p. major, which had no treatment effect (P < 0.05). During differentiation, decreasing Met/Cys concentration caused an increase in early apoptotic cells in both fiber types and no effect on late apoptotic cells except for an increase in the p. major 7.5/24 mg/L of Met/Cys treatment. In general, the viability of the SC was unaffected by the Met/Cys concentration except during proliferation in the p. major 0/0 mg/L of Met/Cys treatment, which increased SC viability. These data demonstrate the effect of nutrition on SC transdifferentiation to an adipogenic lineage and apoptosis, and the effect of fiber type on this response in an in vitro context.

Early posthatch satellite cell (SC) mitotic activity is a critical component of muscle development and growth. Satellite cells are stem cells that can be induced by nutrition to follow other cellular developmental pathways. The objective of the current study was to determine the effect of restricting protein synthesis on the proliferation and differentiation of SC, using variable concentrations of Met and Cys to modulate protein synthesis. Broiler pectoralis major SC were cultured and treated with 1 of 6 different Met/Cys concentrations: 60/192, 30/96 (control), 7.5/24, 3/9.6, 1/3.2, or 0/0 mg/L. The effect of Met/Cys concentration on SC proliferation and differentiation was measured, and myonuclear accretion was measured by counting the number of nuclei per myotube during differentiation. The 30/96 mg/L Met/Cys treatment resulted in the highest rate of proliferation compared with all other treatments by 72 h of proliferation (P < 0.05). Differentiation was measured with Met/Cys treatments only during proliferation and the cultures receiving normal differentiation medium (R/N), normal proliferation medium and differentiation medium with variable Met/Cys (N/R), or both proliferation and differentiation receiving variable Met/Cys treatments (R/R). Differentiation responded in a dose-dependent manner to Met/Cys concentration under all 3 of these treatment regimens, with a degree of recovery in the R/N regimen cells following reinstatement of the control medium. Reductions in both proliferation and differentiation were more pronounced as Met/Cys concentrations were further reduced, whereas increased differentiation was observed under the increased Met/Cys concentration treatment when applied during differentiation in the N/R and R/R regimens. The number of nuclei per myotube was significantly decreased in the severely Met/Cys restricted treatments (P < 0.05). These data demonstrate the sensitivity of pectoralis major SC to nutritional availability and the importance of optimal nutrition during both proliferation and differentiation for maximizing SC activity, which will affect subsequent muscle mass accretion.

The effect of hatch time and subsequent access to feed on intramuscular adipose tissue deposition was studied in the pectoralis major muscle of male Ross 308 broiler chickens. Based on their hatch time chicks were classified as early (EH), midterm (MH), or late (LH) hatchers, with an average incubation duration of 497.7 h for EH, 508.8 h for MH, and 514.5 h for LH birds. Chicks were provided access to feed either immediately at hatch, or 24 h after the conclusion of the hatch window. Expression of the adipogenic regulatory genes peroxisome proliferator-activated receptor gamma (PPARγ), and stearoyl-CoA desaturase (SCD), were measured at the time of hatch, and zero, one, 4, 7, 28, and 40 d. Intramuscular adipocyte cell width and visualization of adipose tissue deposition was observed at 28 and 40 d. Expression of PPARγ was increased in the pectoralis major of LH birds at the time of hatch, zero, and one d. The expression of PPARγ at one and 7 d, and SCD at 7 d were increased in all birds that received delayed access to feed. At 28 d, adipocyte cell width was increased in LH birds with delayed access to feed, compared to EH and MH birds with delayed access to feed and LH birds with immediate access to feed. At 40 d, adipocyte cell width was increased in all birds that received delayed access to feed. Also at 40 d, there was a trend (P = 0.078) for more extensive intramuscular adipose tissue deposition in LH than EH birds, and in birds with delayed access to feed (P = 0.075). These data indicate delayed access to feed increases intramuscular adipose tissue deposition in the pectoralis major muscle, and suggest that hatch time influences this regulation.

The effect of hatch time and the timing of access to feed on growth rate and breast muscle development was assessed in Ross 308 broiler chickens. Chicks were removed from the incubator upon hatching, and classified as early (EH), midterm (MH), or late (LH) hatchers, based on the duration of their incubation. Feed and water were available either immediately at hatch, or 24 h after the conclusion of the hatch period. Hatchling body weight was uniform regardless of hatch time. Subsequently, bodyweight was increased in EH compared to LH birds following immediate access to feed, until 7 d in female, and 14 d in male birds. Relative breast weight was increased until 28 d in birds with immediate access to feed, and also EH and MH birds regardless of access to feed. Pectoralis major muscle morphology and expression of the myogenic regulatory factors myogenic determination factor 1 (MYOD1) and myogenin, and the proteoglycans syndecan-4, glypican-1, and decorin were measured. Myogenin and glypican-1 stimulate satellite cell (SC) differentiation. Glypican-1 expression was unaffected by treatment. A late increase in myogenin expression was observed in MH birds with delayed access to feed, and all LH birds. Syndecan-4 and MYOD1, expressed in proliferating SC, and decorin, which stimulates satellite cell proliferation and differentiation, were variably upregulated in the first wk posthatch in the same birds. These data suggest SC were activated and proliferating, but had reduced differentiation in later hatching and feed deprived birds. Conversely, EH birds with immediate access to feed had maximal myofiber width at 7 d, while fiber width was increased in birds with immediate access to feed compared to those with delayed access to feed through 40 d of age. These results demonstrate that delaying chick access to feed for 24 h upon removal from the incubator will impair muscle growth. Additionally, hatch time influences muscle development, with accelerated muscle growth in EH and MH, compared to LH birds, irrespective of access to feed.

The simultaneous addition of xylanase (5,600 EXU/kg) and phytase (500 FTU/kg) feed enzymes to wheat-based broiler diets was investigated. Starter, grower and finisher diets, with three tiers of nutrient specifications, were fed to 1,440 broiler chicks kept on deep litter from 1-42 days post-hatch, without and with xylanase plus phytase, to determine the effects of diet type and enzyme supplementation on growth performance. The nutrient specifications of type A diets were standard; energy density and protein/amino acid levels were reduced on a least-cost basis to formulate type B diets and further reduced to type C diets. Phosphorus (P) and calcium (Ca) levels were adjusted in supplemented diets. From 1-42 days post-hatch, diet type significantly influenced growth performance. Birds on type C diets had lower growth rates (2,429 vs. 2,631 g/bird; p

Satellite cells (SC) are a multipotential stem cell population responsible for facilitating posthatch muscle fiber hypertrophy. The proliferation and differentiation of SC is sensitive to nutritional regimen, and the SC response to nutrition varies depending upon their muscle type of origin. The objective of the current study was to determine the effect of altering protein synthesis on the expression of several key genes regulating SC activity and the effect of muscle type. Satellite cells isolated from the fast glycolytic pectoralis major and the fast oxidative and glycolytic biceps femoris were studied. These genes included the myogenic regulatory factors myogenic determination factor 1 (MyoD) and myogenin, the cell-membrane associated proteoglycans syndecan-4 and glypican-1, the extracellular matrix proteoglycan decorin, and the transcription factor paired box 7. Protein synthesis potential varied by the concentration of the sulfur amino acids Met and Cys during SC proliferation and differentiation. The SC were cultured and treated with 1 of 6 Met/Cys concentrations: 60/192, 30/96 (control), 7.5/24, 3.0/9.6, 1.0/3.2, or 0/0 mg/L. A consistent pattern of gene expression emerged following Met/Cys manipulation as increasing reductions in mRNA expression for all genes were observed as Met/Cys concentration decreased, whereas increased Met/Cys concentration caused either no change or had a small negative effect on mRNA expression. Reduced paired box 7 expression would limit myogenic specification of SC, whereas decreased myogenic regulatory factor expression would affect subsequent myogenic development of the SC. Decreased levels of decorin affect SC response to growth factors like myostatin and transforming growth factor β, and extracellular matrix organization. These data highlight the importance of nutrition on the expression of genes critical for satellite cell activation, proliferation and differentiation, and growth factor signal transduction.

The effect of primary infection and subsequent challenge with Eimeria tenella on interferon-gamma(IFN-gamma) production in the spleen and caeca of Light Sussex chickens was assessed. The ability of splenocytes to proliferate and produce IFN-gammain response to mitogen stimulation ex vivo was determined. Differences in the kinetics of IFN-gammaproduction suggested that the spleens of infected birds contain a subpopulation of T cells, primed to produce IFN-gamma, which migrate from the spleen in response to a secondary infection. IFN-gammamRNA expression was detected by hybridization of an anti-sense chicken IFN-gammariboprobe to splenic sections from infected birds and caecal sections from challenged birds. Hybridization was to T-cell areas in the spleen, and to cells in the lamina propria and intraepithelial compartments of the caecum. This is the first direct demonstration of IFN-gammaexpression in chickens at the site of E. tenella infection, and also the first indication that IFN-gammamay be involved in the immune response to challenge.

Defence of the intestinal mucosal surface from enteric pathogens is initially mediated by secretory IgA (SIgA). As oral immunization of non-replicating antigen induces minimal SIgA antibody titers, novel immunization strategies which selectively induce mucosal immune responses in mammals are now being assessed in chickens. The strategies reviewed include the route of antigen delivery, the incorporation of antigenic components in delivery vehicles, the inclusion of immunomodulators in the vaccine formula or in the diet, and manipulation of intestinal microflora. The differences in anatomical organization and immunological mechanisms between birds and mammals must be considered when manipulating avian intestinal immunity with the latest immunotechnologies developed for mammals. Our knowledge of the function and functioning of the avian mucosal system is discussed. Progress in our understanding of this system, the location of precursor IgA B cells and antigen sampling by these sites is not as advanced as knowledge of the mammalian system, highlighting the need for ongoing research into the avian application of novel vaccination strategies.

Conventional methods of parenteral immunization with killed bacterin vaccines have met with limited success in protecting the avian intestinal mucosa from pathogens such as Salmonella typhimurium. For mucosal vaccines to be successful they must be evaluated for their ability to stimulate local secretory immunoglobulin (SIgA) at the mucosal surface, which acts as the first line of defense against invading pathogens. Previously we have demonstrated the ability of i.p. immunization with nonreplicating antigen in an appropriate adjuvant to induce a primary immune response, which, after an oral booster immunization, stimulates enhanced intestinal IgA responses in chickens. In the experiments reported here we have applied this immunization protocol to vaccinate against S. typhimurium in chickens, and examined the protection provided against subsequent S. typhimurium challenge by placing vaccinated birds on seeded litter with cohabitant infected birds. Immunized+challenged birds displayed delayed onset of S. typhimurium infection, both at the mucosal surface and within the reticuloendothelial system. Elevated anti-S. typhimurium IgG and IgA titers were detected in serum after vaccination, which markedly increased after challenge, to levels higher than in control+challenged chickens. Anti-S. typhimurium IgA in bile and intestinal scrapings supernatant was also higher in the immunized+challenged birds than in the control+challenged birds 15 d after challenge. This study illustrates the potential for i.p. vaccination to induce a mucosal immune response to S. typhimurium in chickens, which, in the challenge model employed here, provided partial protection against intestinal challenge with the same pathogen and was reflected in deferred onset of bacterial infection and shedding.

The effect of feeding diets of variable methionine concentration on breast muscle development was assessed in Ross 308 broiler chicks. Four isonitrogenous and isoenergetic starter diets were formulated to contain 7.8, 5.9, 4.6, and 3.4 g methionine/kg diet, and were provided for the first 7 days post-hatch. At 7 days of age all birds were placed on an industry standard starter diet with 5.9 g methionine/kg diet until 14 days, and then provided standard broiler grower (until 28 days) and finisher (until 42 days) diets. Birds were weighed periodically throughout the study and feed intake and feed conversion ratio were determined. Ten birds per treatment were sacrificed and weighed on 0, 1, 4, 7, 14, 21, 28, 35, and 42 days. The pectoralis major (breast muscle) was then removed from the carcass and weighed. Samples of breast muscle were collected for genetic and histological analysis. Expression of the myogenic marker genes, myogenic differentiation factor 1 and myogenin, which regulate satellite cell activity, and the adipogenic marker gene, peroxisome proliferator-activated receptor gamma (PPARγ), was measured. Histological assessment of breast muscle morphology and fat deposition morphology was also performed. No effect of dietary treatment was observed on body or breast muscle weight, feed intake or feed conversion ratio. Marker gene expression was also similar in all treatment groups, except for PPARγ. Significantly higher expression of PPARγ was observed at 0 days in the 5.9 g methionine/kg diet treatment, before dietary treatments were provided. Expression of PPARγ did not differ among treatment groups on any subsequent day. Methionine dietary treatment had no effect on the morphological structure of the breast muscle, or intramuscular fat deposition. These results suggest that under the conditions of this study, satellite cell activity in the early post-hatch chick, and subsequent muscle development, were not responsive to the variable methionine manipulations tested in the pre-starter period.

Bursectomized chicks received lymphocyte single cell suspensions harvested from the bursa of Fabricius (BF), ileal lymphoid aggregate (ILA), caecal tonsils (CT), spleen and peripheral blood. Four days after cell transfer, repopulation of the duodenal and CT lamina propria in age-matched recipient bursectomized chickens with IgA-secreting plasma cells was determined. The results indicate the highest level of reconstitution with cells derived from BF, but substantial numbers of IgA-secreting plasma cells were also observed in a number of birds that received lymphocytes originating from the ILA and CT.

The effect of dietary vitamin E on immunoglobulin A (IgA) antibody production, which acts as the first line of defence at the intestinal mucosa, has not been evaluated in chickens. In the present study the impact of the inclusion of supplementary levels of vitamin E to the diet, on total and antigen-specific IgA antibody titres, T-cell subsets and Ia+ cells, was assessed. From hatching, chickens received a maize-based diet which was supplemented with either 25, 250, 2500 or 5000 mg dl-α-tocopherol acetate/kg. Primary immunisation with tetanus toxoid (T. toxoid) emulsified in a vegetable oil-in-water adjuvant was administered by the intraperitoneal route at 21 d of age. At 35 d of age all birds received an oral booster vaccination of T. toxoid. Significantly higher total IgA antibody titres were present in the day 42 intestinal scrapings of birds receiving the 5000 mg/kg vitamin E-supplemented diet (VESD) (P=0·05) and a notable increase was observed in birds receiving the 250 mg/kg VESD (P=0·06). At days 21 and 42 total serum IgA antibody titres of birds receiving the 250 mg/kg VESD was significantly higher (PP=0·02). Both of these groups also demonstrated increased titres of anti-T. toxoid IgA in the serum at day 42. Birds receiving the 250 mg/kg VESD exhibited a notable increase in the percentage of T-helper cells and Ia+ cells in peripheral blood on day 26. The results illustrate the potential for some levels of dietary vitamin E supplementation to act as an immunomodulator of total and antigen-specific IgA antibody.

Experimental infection of chickens with controlled quantities of Salmonella typhimurium is often achieved by administration of a single oral inoculum of live bacteria to caged chickens. However, this method is a poor simulation of the natural process of S. typhimurium infection in the field, making the practical application of results obtained under such conditions tenuous. This experiment was designed to evaluate the use of horizontal transmission for the challenge/infection of chickens with S. typhimurium with the expectation that it may more closely resemble the natural situation and, therefore, the in-field physiological response of chickens. Further, the experiment allowed for comparison of both the kinetics and magnitude of the mucosal immune response following each mode of challenge by exposing the chickens to challenge by placing them on litter with S. typhimurium-infected seeder birds. Overall, birds challenged via seeded litter exhibited a slower rate of infection and a more gradual increase in serum antibody production compared with birds receiving a single oral inoculum.

1. Systemic and intestinal antibody titres were measured in chickens following subcutaneous, intraperitoneal (i.p.), oral (p.o.) and combined i.p./p.o. administration of antigen, in soluble, emulsified or microparticulate form. Antigens tested included keyhole limpet haemocyanin (KLH), killed Campylobacter jejuni whole cells and purified campylobacter flagellin protein. 2. The effect of immunisation with purified flagellin protein or with killed C. jejuni whole cells in reducing intestinal colonisation was assessed. The ability of newlyhatched chicks to respond to immunisation was limited, possibly because of the immaturity of the immune system rather than maternal suppression of an immune response. Only 5 to 13 birds that were first immunised when 1-d-old with KLH showed a systemic response, even after 4 immunisations, whereas 10 of 11 birds that were first immunised at 24 d-old responded systemically. 3. In an immunisation and challenge experiment, birds that were immunised twice intraperitoneally, at 16 and 29 d-old, with killed C. jejuni whole cells, had fewer C. jejuni, in the caecal contents than unimmunised control birds. This reduction in intestinal colonisation, to less than 2% of bacterial numbers in control birds, was associated with an increase in specific IgG in intestinal secretions. There was no significant increase in specific IgA or IgM in intestinal secretions following immunisation and challenge. 4. These results indicate that immunisation can reduce the level of intestinal infection with C. jejuni. The protection may be enhanced by developing improved methods of immunisation that stimulate production of increased titres of specific antibody in intestinal secretions, particularly specific IgA antibody.

A study was designed to assess the impact of water-soluble proteins and peptides extracted from meat and bone meal (MBM) on broiler chick performance, following their oral delivery during the early post-hatch period. Proteinaceous material was fractionated by size exclusion filtration into weight ranges of 100 kDa (Fraction 3; 0.8 mg protein/mL), which formed the three protein fraction treatments. A total of 1 mL of each of the respective preparations was delivered orally via gavage over 4 days (0.25 µL each day) to Cobb broiler hatchlings. Three control groups: control–unhandled, control–phosphate-buffered saline and control–handled were also included. Chicks were grown to 30 days of age. Feed intake, chick weight gain and feed conversion ratio were determined from day old through to 29 days of age. On Days 10, 16, 23 and 30, the weight of the breast and the small intestine was determined from 10 birds/treatment. For all parameters measured there was no interaction between experimental week and protein fraction treatment. Chicks receiving Fraction 2 had a statistically significant increase in feed intake and weight gain (P = 0.012) compared with the control–unhandled chicks. Chicks receiving Fraction 2 also demonstrated a numerically higher final bodyweight. Mass spectrometric analysis of all three fractions revealed that they each contained a wide array of proteinacious material. The results of this study suggests the likelihood that protein or protein-derived fragment components within the 3–100 kDa molecular weight range of MBM can generate improvements in broiler chick production, and thus promote the need for further research to identify the specific protein(s) responsible for the observed positive growth effects.

The limited success in stimulating protective immunity in the intestine by traditional vaccination approaches has led to the search for novel strategies to improve intestinal immunity. In mammalian species we have demonstrated that whereas oral immunization produces poor intestinal responses, immunization by the intraperitoneal route using appropriate formulations is an effective means of priming the intestinal lymphoid tissue for an enhanced mucosal response to orally administered antigen. In the experiments reported here we extend these findings to chickens and compare the intestinal IgA response to a non-replicating antigen administered by the oral route alone or in oil emulsion formulations administered subcutaneously or intraperitoneally with or without oral boosting. Despite repeated daily dosing, oral immunization of unprimed animals produced only a small intestinal response. However, priming by the intraperitoneal route with antigen in either Freund's adjuvant or a biodegradable oil-in-water emulsion (Auspharm adjuvant) resulted in a markedly improved response, especially when additional oral boosting was given. The data also demonstrate that Quil A, a saponin derivative, further boosts the response when co-administered with the oil emulsion intraperitoneally. This study illustrates the efficacy of systemic immunization by the intraperitoneal route in priming for an intestinal IgA response in chickens.

Diseases which affect mucosal surfaces cause considerable mortality and morbidity. New vaccine technologies are now available which justify a reappraisal of oral delivery not only for infectious disease control but also to control mucosal physiological processes such as fertility. Biodegradable microspheres have been investigated for their use as an oral delivery vehicle in rats using a recombinant antigen derived from fox sperm. Unencapsulated antigen administered in saline by the oral route produced a negligible response although an improved response was obtained if administered directly into the duodenum. This response was considerably enhanced if Peyer's patch (PP) priming was performed by direct injection of antigen in Freund's complete adjuvant (FCA) prior to intraduodenal (ID) delivery. In contrast, microencapsulated antigen given orally produced a substantial response, which was predominantly IgA specific, and almost equal in magnitude to that obtained by PP priming and ID boosting with native antigen. Direct ID delivery produced a similar response but when PP were primed with microencapsulated antigen in FCA the response to ID boosting was greater than with any of the other protocols investigated. These data demonstrate the efficacy of biodegradable microspheres in producing an IgA antibody response following oral vaccination.

Experimental infection of chickens with controlled quantities of Salmonella typhimurium is often achieved by administration of a single oral inoculum of live bacteria to caged chickens. However, this method is a poor simulation of the natural process of S. typhimurium infection in the field, making the practical application of results obtained under such conditions tenuous. This experiment was designed to evaluate the use of horizontal transmission for the challenge/infection of chickens with S. typhimurium with the expectation that it may more closely resemble the natural situation and, therefore, the in-field physiological response of chickens. Further, the experiment allowed for comparison of both the kinetics and magnitude of the mucosal immune response following each mode of challenge by exposing the chickens to challenge by placing them on litter with S. typhimurium-infected seeder birds. Overall, birds challenged via seeded litter exhibited a slower rate of infection and a more gradual increase in serum antibody production compared with birds receiving a single oral inoculum.