Differential expression of iodothyronine deiodinase type 2 in growth plates of chickens divergently selected for incidence of tibial dyschondroplasia

Tibial dyschondroplasia (TD) is a genetic leg defect in broilers with a lesion of avascular, non-calcified cartilage below the growth plate of the proximal tibiatarsus. This disease is considered to result from the inability of chondrocytes to undergo terminal differentiation. Thyroid hormones are required for chondrocyte differentiation. The thyroid gland produces and secrets mostly L-thyroxine or T4 and T4 plays most of its biological activities through conversion to triiodothyronine or T3 in local tissues by iodothyronine deiodinases type 1 or type 2, which are tissue specific. In this study, no differences were found in the plasma concentrations of total T3 and T4 between two chicken lines divergently selected for the incidence of TD. Plasma T4 was higher than T3, especially in older chickens. Younger birds had much higher T3 than older birds, but there were no significant age differences in T4. The expression level of deiodinase type 2 in the growth plates of broilers with TD was one-eighth of those birds without the disease. The expression levels of deiodinase type 2 (DIO2) in commercial broilers without the disease were much higher than those with TD and lower than those without the disease in the susceptible and resistant lines, respectively. These results indicate that the inadequate expression of DIO2 in the growth plates contributes to the pathogenesis of TD in broilers and that TD is a tissue-specific hypothyroidism.     

Perches and elevated platforms in commercial broiler farms: use and effect on walking ability,incidence of tibial dyschondroplasia and bone mineral content

Modern fast-growing broilers spend excessive periods resting and their activity further decreases with age. Inactivity has been suggested to increase impaired gait and the incidence of leg disorders. Tibial dyschondroplasia (TD) is a common leg pathology in broilers. A more complex environment might facilitate more activity and improve leg health. Perches or elevated platforms bring variety to broilers’ environment and could motivate more locomotion. This study examined the impact of perches and elevated platforms on walking ability, the occurrence of TD and level of bone ash and mineral contents. The investigation was performed on four commercial broiler farms throughout six consecutive batches with platforms and four to five with perches. On each farm at least two separate houses were included, enabling the comparison of furnished flocks to control flocks during each batch. Plastic slats with ramp access elevated by 30 cm or wooden perches of 10 and 30 cm height were offered in the furnished house. Farmers recorded the platform and perch usage twice a week with a five-point scale. Gait was scored before slaughter on a six-point scale according to the Welfare Quality® assessment protocol for poultry. The severity of TD was determined using a four-point scale on farm from all birds gait scored as 3 and at slaughter from 200 birds/flock. Farmers estimated 50% to 100% of the platforms to be occupied in all flocks throughout the entire growing period. Only single birds were perching, thus perch structures were constantly evaluated to be empty. Due to the low use, the perch-equipped houses were excluded when analysing bone content, walking ability and TD. On average, 30% of the tested birds exhibited gait score ⩾3. Younger scoring age resulted in a lower mean gait score and a lower percentage of scores 3 and 4 to 5. Overall, 2.3% of the birds examined at slaughter and 3.5% of the birds with gait score 3 were affected by TD. Leg health was better in birds with access to platforms: mean gait score, the percentage of birds scoring 3, and TD percentage and severity were lower in birds in platform-equipped houses. Elevated structures such as platforms, offering additional possibilities for locomotion to broilers seem to improve their leg health.     

TDP-1, a toxic component causing tibial dyschondroplasia in broiler chickens, and trichothecenes from Fusarium roseum 'Graminearum'

Fusarium roseum 'Graminearum' was isolated from overwintered oats in Alaska and was tested for its ability to cause tibial dyschondroplasia (TDP) in broiler chickens. The water-soluble fraction was tested and found to cause TDP. In addition, diacetoxyscirpenol and 7-hydroxydiacetoxyscirpenol were identified in the acetonitrile fraction of the extracts and caused mild mouth lesions in chickens. Six major water-soluble components were purified by thin-layer chromatography and tested for toxicity to chick embryos. One of the six components, called TDP-1, was found to be lethal to chick embryos. There was a 100% incidence of TDP in chickens fed a diet containing 75 ppm (wt/wt) of pure TDP-1, thus establishing the cause and effect relationship between TDP and TDP-1. Analyses by thin-layer chromatography and mass spectrometry revealed that TDP-1 is polar and ninhydrin positive, exhibits fluorescence with UV irradiation, and is a nitrogen-containing component with an empirical formula of C15H20N2O4.     

TDP-1, a toxic component causing tibial dyschondroplasia in broiler chickens, and trichothecenes from Fusarium roseum 'Graminearum'.

Fusarium roseum 'Graminearum' was isolated from overwintered oats in Alaska and was tested for its ability to cause tibial dyschondroplasia (TDP) in broiler chickens. The water-soluble fraction was tested and found to cause TDP. In addition, diacetoxyscirpenol and 7-hydroxydiacetoxyscirpenol were identified in the acetonitrile fraction of the extracts and caused mild mouth lesions in chickens. Six major water-soluble components were purified by thin-layer chromatography and tested for toxicity to chick embryos. One of the six components, called TDP-1, was found to be lethal to chick embryos. There was a 100% incidence of TDP in chickens fed a diet containing 75 ppm (wt/wt) of pure TDP-1, thus establishing the cause and effect relationship between TDP and TDP-1. Analyses by thin-layer chromatography and mass spectrometry revealed that TDP-1 is polar and ninhydrin positive, exhibits fluorescence with UV irradiation, and is a nitrogen-containing component with an empirical formula of C15H20N2O4.     

Vocalisation sound pattern identification in young broiler chickens

In this study, we describe the monitoring of young broiler chicken vocalisation, with sound recorded and assessed at regular intervals throughout the life of the birds from day 1 to day 38, with a focus on the first week of life. We assess whether there are recognisable, and even predictable, vocalisation patterns based on frequency and sound spectrum analysis, which can be observed in birds at different ages and stages of growth within the relatively short life of the birds in commercial broiler production cycles. The experimental trials were carried out in a farm where the broiler where reared indoor, and audio recording procedures carried out over 38 days. The recordings were made using two microphones connected to a digital recorder, and the sonic data was collected in situations without disturbance of the animals beyond that created by the routine activities of the farmer. Digital files of 1 h duration were cut into short files of 10 min duration, and these sound recordings were analysed and labelled using audio analysis software. Analysis of these short sound files showed that the key vocalisation frequency and patterns changed in relation to increasing age and the weight of the broilers. Statistical analysis showed a significant correlation (P<0.001) between the frequency of vocalisation and the age of the birds. Based on the identification of specific frequencies of the sounds emitted, in relation to age and weight, it is proposed that there is potential for audio monitoring and comparison with ‘anticipated’ sound patterns to be used to evaluate the status of farmed broiler chicken.     

Effect of match or mismatch of maternal-offspring nutritional environment on the development of offspring in broiler chickens

In mammals, maternal food restriction around conception and during pregnancy results in low birth weight and an adjusted growth trajectory of offspring. If, subsequently, the offspring are born into a food-abundant environment, they are at increased risk of developing obesity, type 2 diabetes, hypertension and renal dysfunction. Here, we show similar effects of maternal undernutrition on hatch weight, growth and fat deposition in offspring of birds (domestic chicken). Both mothers and offspring were fed either ad libitum or restricted in a two-by-two factorial design, resulting in two matched and two mismatched maternal-offspring nutritional environments. Offspring of ad libitum mothers grew heavier than those of restricted mothers, possibly due to the larger muscle mass. Ad libitum-fed offspring, especially females, of restricted mothers were lighter at hatch, and were heavier and had more abdominal fat at 6 weeks of age than daughters of ad libitum-fed mothers. These results suggest a common mechanism in mammals and birds in response to a mismatch in the maternal-offspring nutritional environment. They also indicate that the common practice of restrictive feeding of the broiler breeders and subsequent ad libitum feeding of the broilers may result in reduced growth and increased abdominal fat as compared to broilers of less restricted broiler breeders.     

Three new MHC haplotypes in broiler breeder chickens

Six distinct serotypes of the chicken B blood group system (which encodes the major histocompatibility complex) were identified in a commercial broiler breeder line (Line C). The B serotypes were compared by B-G restriction fragment length polymorphism (RFLP) analysis, allele-specific PCR typing test for B-LBII family genes and nucleotide sequence analysis of expressed B-F and B-LBII family genes. The results indicated the existence of seven distinct B haplotypes. Nucleotide sequence analysis demonstrated that three of the Line C haplotypes encode new B-F and B-LB alleles.     

Feeding graded levels of dried Sea buckthorn (Hippophaes rhamnoides) berries to broiler chickens

The aim of this study was to assess the effects of graded levels (0, 3, 6, 9 and 12 g/kg) of dry Sea buckthorn (SB) berries on growth performance, gastrointestinal tract (GIT) development, jejunal histomorphology, bird antioxidant status and caecal short-chain fatty acid concentration when fed to female Ross 308 broiler chickens. In addition, expression of cytokine biomarker genes in the jejunum was evaluated. The five experimental diets were fed from 7 to 21 days age to 8 pens (two birds in each) following randomisation. Feeding SB did not influence bird growth performance (p > .05). There was a linear decrease in butyric, acetic and valeric acid concentrations in caecal digesta (p < .05) and a decrease (p < .05) in crypt depth. The expression of IFNG and CD40LG responded quadratically (p < .05), peaking at 6–9 g/kg dietary inclusion of SB, respectively. Other studied variables were not affected by dietary SB inclusion (p > .05). Feeding dry SB berries up to 12 g/kg of diet did not improve the zootechnical variables of healthy commercial-strain broilers in this study.     

An evaluation of potential dustbathing substrates for commercial broiler chickens

Provision of an appropriate dustbathing substrate may allow broiler chickens to satisfy a natural motivation and give them an opportunity to exercise. The main aim of this study was to evaluate the extent to which different substrates promote dustbathing behaviour in broilers. The trial was replicated over three production cycles in one commercial broiler house, with ~22 000 Ross broilers housed per cycle. The birds were provided with access to five experimental substrates from day 10 of the 6-week production cycle. The substrates included the following: (1) peat (P), (2) oat hulls (OH), (3) straw pellets (SP), (4) clean wood shavings (WS), and (5) litter control (C). The substrates were provided in 15 steel rings (1.1 m in diameter, three rings per substrate) dispersed throughout the house. The level of occupancy of the rings, behaviours performed in each substrate, and the effect of ring position (central or edge of house) were assessed in weeks 3, 4, 5 and 6 using scan sampling from video footage. Where substrates successfully promoted dustbathing, the length and components of the bouts (including number of vertical wing shakes and ground pecks) were also assessed. Results showed that birds used P significantly more than the remaining substrates for dustbathing (P<0.001). Oat hulls were the second most preferred substrate for dustbathing, with significantly more birds dustbathing in the OH compared with SP, WS and C (P<0.001). The least sitting inactive was also seen in the P and OH rings compared with the SP, WS and C (P<0.001). The highest levels of foraging were recorded in the P, OH and WS compared with SP and the C. Position of the rings did not affect the types of behaviours performed in any substrate, although overall more birds were counted in the central compared with edge rings (P=0.001). More detailed information on dustbathing behaviour was only recorded in the P and OH treatments, and there were no differences in the length of dustbathing bout, or components of the bout between them (P>0.05). The use of OH is likely to be more environmentally sustainable than that of P, and our results suggest that this substrate is relatively successful in promoting dustbathing. However, a preference was still observed for P and further work should investigate whether other suitable substrates could better reflect its qualities.     

Molecular characterization of major histocompatibility complex (B) haplotypes in broiler chickens

In Leghorn (laying) chickens, susceptibility to a number of infectious diseases is strongly associated with the major histocompatibility ( B ) complex. Nucleotide sequence data have been published for six class I ( B-F ) alleles and for class II ( B-Lβ ) alleles or isotypes from 17 Leghorn haplotypes. It is not known if classical B-L or B-F alleles in broilers are identical, at the sequence level, to any Leghorn alleles. This report describes molecular and immunogenetic characterization of two haplotypes from commercial broiler breeder chickens that were originally identified by serology as a single haplotype, but were differentiated serologically in the present work. The two haplotypes, designated B ^A4 and B ^A4variant, shared identical B-G restriction fragment length polymorphism patterns, but differed in one B-Lβ fragment that cosegregated with the serological B haplotype. Furthermore, the nucleotide sequences of the highly variable exons of an expressed B-LβII family gene and B-F gene from the two haplotypes were markedly different from each other. Both the B-LβII family and B-F gene sequences from the B ^A4 haplotype were identical to the sequences obtained from the reference B ²¹ haplotype in Leghorns; however, in the B ^A4 haplotype the B-Lβ ²¹ and B-F ²¹ alleles were in linkage with B-G alleles that were not G ²¹. The nucleotide sequences from B ^A4variant were unique among the reported chicken B-LβII family and B-F alleles.     

Associations of myostatin gene polymorphisms with performance and mortality traits in broiler chickens

Myostatin is a negative regulator of skeletal muscle growth. We evaluated effects of myostatin polymorphisms in three elite commercial broiler chicken lines on mortality, growth, feed conversion efficiency, ultrasound breast depth, breast percentage, eviscerated carcass weight, leg defects, blood oxygen level, and hen antibody titer to infectious bursal disease virus vaccine. Progeny mean data adjusted for fixed and mate effects and DNA from 100 sires per line were used. Single nucleotide polymorphisms (SNPs) of the myostatin gene segregating in these lines were identified by designing specific primers, amplifying individual DNA in each line by polymerase chain reaction, cloning, sequencing and aligning the corresponding products. Individual sires were genotyped for five identified SNPs which contributed to eight haplotypes. Frequencies of SNP alleles and haplotypes differed between lines. Using the allele substitution effect model, the myostatin SNPs were found to have significant (P < 0.031) associations with growth, mortality, blood oxygen and hen antibody titer to infectious bursal disease virus vaccine, although the associations were not often consistent across lines. These results suggest that the myostatin gene has pleiotropic effects on broiler performance.     

MX1 exon 13 polymorphisms in broiler breeder chickens and associations with commercial traits

The Mx1 gene in mice is induced by type I interferons and is the major determinant of resistance to influenza virus and related orthomyxoviruses. It has been previously shown that a SNP in exon 13 of the chicken MX1 gene determines differential antiviral activity of the protein. We evaluated this SNP and two additional SNPs in elite broiler lines by PCR amplification and sequence analysis. Associations between MX1 exon 13 SNPs and several traits of economic interest were evaluated. Significant associations were found between the SNP determining antiviral activity and mortality in one line and leg defects in another line.     

Effect of different selenium source (sodium selenite and selenium yeast) on broiler chickens

A feeding experiment was carried out to compare the effects of supplementing a poultry meal-based diet with selenium as sodium selenite or selenium yeast on broiler chickens. Three groups with three replicates of broiler chickens (mean weight 710 ± 5.3 g) were given a basal diet either unsupplemented (control) or supplemented with 0.2 mg Se kg⁻¹ as sodium selenite (trial 1) or selenium yeast (trial 2) respectively, for 21 days. There was significant difference (P<0.05) in Feed Conversion Ratio (FCR) of trials 1 and 2 compared with the control. However, there were no significant differences (P>0.05) in FCR between trials 1 and 2. Final weight, survival rate and Daily Gain (DG) were not affected by the dietary Se source. Chickens fed the basal diet showed lower (P<0.05) selenium content in muscle, kidney, liver and pancreas compared to that fed selenium supplements (trials 1 and 2). Furthermore, trial 2 showed the highest value (P<0.05) among these treatments. However, there was no significant difference (P>0.05) in muscle selenium content of chickens between trials 1 and 2. Glutathione peroxidase (GSH-Px) activities in broiler chickens plasma and liver of all selenium treatment groups (trials 1 and 2) were significantly different (P<0.05) from that of the control. The GSH-Px activity in plasma was higher (P<0.05) in trial 2 compared with trial 1 and the control. However, there was no difference (P>0.05) in hepatic glutathione peroxidase between trials 1 and 2 although the average value of GSH-Px activity in trial 2 presented the trend of increase.     

Genotypic variability at the major histocompatibility complex (B and Rfp-Y) in Camperos broiler chickens

Evidence for the importance of major histocompatibility complex (MHC) genotype in immunological fitness of chickens continues to accumulate. The MHC B haplotypes contribute resistance to Marek's and other diseases of economic importance. The Rfp-Y, a second cluster of MHC genes in the chicken, may also contribute to disease resistance. Nevertheless, the MHC B and Rfp-Y haplotypes segregating in broiler chickens are poorly documented. The Camperos, free-range broiler chickens developed in Argentina, provide an opportunity to evaluate MHC diversity in a genetically diverse broiler stock. Camperos are derived by cross-breeding parental stocks maintained essentially without selection since their founding. We analysed 51 DNA samples from the Camperos and their parental lines for MHC B and Rfp-Y variability by restriction fragment pattern (rfp) and SSCP typing methods for B-G, B-F (class Ia), B-Lbeta (class II) and Y-F (class Ib) diversity. We found evidence for 38 B-G genotypes. The Camperos B-G patterns were not shared with White Leghorn controls, nor were any of a limited number of Camperos B-G gene sequences identical to published B-G sequences. The SSCP assays provided evidence for the presence of at least 28 B-F and 29 B-Lbeta genotypes. When considered together B-F, B-L, and B-G patterns provide evidence for 40 Camperos B genotypes. We found even greater Rfp-Y diversity. The Rfp-Y class I-specific probe, 163/164f, revealed 44 different rfps among the 51 samples. We conclude that substantial MHC B and Rfp-Y diversity exists within broiler chickens that might be drawn upon in selecting for desirable immunological traits.     

Non-intrusive tracking of commercial broiler chickens in situ at different stocking densities

Unlike several other farm animal species, the broiler chicken remains unprotected by species-specific legislation. The densities at which broilers should be kept is a highly contentious issue—some studies have demonstrated increased welfare problems at higher densities, whilst a few others have, contrary to expectations, suggested that broilers may actually find crowds of other birds attractive. A tracking method was developed and used to provide an insight into the social preferences of commercial broiler chickens in situ—inside commercial, closed-system broiler houses. The aim was to simultaneously assess the relative impact of global measures of density, such as target and actual stocking densities and local measures of the social environment on the behaviour and route taken to feed by focal birds. Birds were tracked inside 20 commercial broiler houses across the UK. Results from this study show that stocking density per se seems to have little direct effect on the individual behaviours of focal broiler chickens. However, there may still be an indirect effect of stocking density on broiler behaviour, mediated through the local social environment.     

Associations of BF2 alleles with antibody titres and production traits in commercial pure line broiler chickens

Three commercial broiler pure lines were evaluated for associations of sire BF2 (major histocompatibility complex class I) alleles with progeny phenotypic traits. Significant BF2 associations with a subset of traits were observed in two lines. The BF2*21 allele was positively associated with antibody titre to infectious bursal disease virus in both lines. Other associations were line-specific.     

Mechanism and cost of synchronous hatching

1. Synchrony in the timing of births is thought to have evolved as a general predator avoidance strategy. In turtles, synchronous hatching may facilitate group emergence from the nest, which in turn, may limit predation by diluting an individual's risk of predation or by simply swamping predators upon emergence. However, synchronizing hatching should not be easily achieved in natural nests because of thermal gradients affecting developmental rates.
2. We evaluated pipping synchrony in the painted turtle ( Chrysemys picta ), where the drive to hatch synchronously may be reduced, because in many populations, hatching and emergence are dissociated through hatchlings overwintering within a nest.
3. We also assessed developmental mechanisms through which synchrony occurs and explored potential trade-offs between pipping synchrony and individual fitness by determining potential short and long-term neuromuscular developmental costs to hatching prematurely. These data were also used to develop a theoretical model to determine how differential embryonic maturation rates affect hatching synchrony.
4. Underdeveloped embryos pipped much earlier than expected and at similar times to their more advanced sibs. In addition, a trade-off between hatching synchronously and neuromuscular development (motility) was evident several days after hatching and up to 9 months later, after the overwintering period.
5. Synchronous hatching is an ancestral trait in turtles, but its relevance today is not solely for predator avoidance in all species.
6. Abiotic factors during incubation (e.g. temperature regime and moisture) have long-term effects on reptiles during ontogeny but it is also clear that incubation behaviour is major factor for the persistence of these developmental costs.