Corticosterone in feathers: Inter- and intraindividual variation in pullets and the importance of the feather type.

Measuring corticosterone concentrations in feathers of poultry may be suitable to determine birds' exposure to stress. It is thinkable, that in laying hens such information could be helpful as an animal welfare indicator to evaluate adverse husbandry conditions and to predict the risk of developing behavioral disorders, such as feather pecking and cannibalism. Yet, there are some fundamental issues which remain unclear. Therefore, the objective of the current pilot study was to examine the inter- and intraindividual variation of pullets at the end of the rearing period, when most of the feathers are fully grown and animals are reaching sexual maturity. Flight feathers from both wings (n = 4), the tail (n = 2 - 3), and body feathers (n = 1 pool of 3 - 5 feathers) were taken from pullets (n = 10), genetics Lohmann Brown, at an age of 19 weeks who were reared in the same flock (N = 728). Corticosterone analysis was performed applying a validated protocol for laying hens. Results indicate not only high intraindividual, but also high interindividual variation. Mean over all samples was 75.2 pg/mg (± 38.58 pg/mg, n = 76), showing higher intraindividual variation (between feather types; SD: 23.75 pg/mg - 49.38 pg/mg; n = 10 pullets) than interindividual variation (within feather types; SD: 11.91 pg/mg - 49.55 pg/mg; n = 6 feather types). The variation between different feather types within one bird was higher than the variation within one feather type between different birds, indicating that birds a) may respond differently when exposed to stressors and b) corticosterone measurements should be done with the same feather type.

Foot Pad Health as Part of On-Farm-Monitoring in Turkey Flocks.

Currently, there is no consistent approach to on-farm and post-mortem foot pad (FP) assessment in turkey husbandry in sampling of both feet, sample sizes of birds and scoring schemes during the production period. Therefore, in a field study, 11,400 turkeys, i.e., 22,800 feet, were macroscopically scored at 4-week intervals, 60 birds per flock per date, in accordance with the scale system of Hocking et al. (1). Spearman's rho was calculated between the foot pad dermatitis (FPD) score of both feet of an individual turkey. Sample size for FPD monitoring was calculated for several flock sizes, considering expected FPD prevalence and the error and confidence level (α = 0.01, 0.05, 0.1). To compare macroscopic to histological findings, ten excised FPs were histopathologically investigated by hematoxylin & eosin staining. To align manual macroscopic FPD evaluations with a technical system, 20 photographic images of FPD were measured using the ImageJ program. The scores of both feet of an individual turkey correlated between r = 0.252 and r = 1.000. Thus, both feet of a bird should be monitored, while the worse foot should be evaluated. As an exemplary sample size for on-farm FPD assessment, 77 turkey poults were calculated in a flock of 4,000 birds with an expected FPD prevalence of 40% and α = 0.1. The sample size of monitored birds within a flock should differ and depend on flock size and expected FPD prevalence. Histopathological findings showed normal and non-affected structures of a macroscopic Score 0 and a moderate ulcer of the macroscopic Score 1 and Score 2. The applied assessment scheme should distinguish first alterations and scar tissue as separate scores to differentiate the need for management intervention vs. the success of management measures that were already implemented. FPD affected areas were given lower Scores and assessed to be healthier when evaluated by an image system, compared to a manual assessment. Furthermore, with regard to an increase in camera-based assessments, the boundary of the metatarsal pad needs to be clarified. In conclusion, a new scoring system is required, as the size of the FP cannot be clearly defined and different tissue textures, as well as valid sample sizes are not currently sufficiently considered.

Precision, repeatability and representative ability of faecal egg counts in Heterakis gallinarum infected chickens.

This study investigated whether a precise and repeatable quantification of Heterakis gallinarum egg excretion, which considerably reflects the actual worm burdens, can be achieved based on collection of the daily total amount of faeces from chickens. Three-week-old birds (N=64) were infected with 200 embryonated eggs of H. gallinarum, and placed into individual cages 3 wk after infection for 5 wk to collect daily faeces (N=2240). The total daily faeces was mixed and a randomly taken sample per bird was analyzed to estimate the numbers of eggs per gram of faeces (EPG) and total number of eggs excreted within 24h (EPD). A total of 235 daily faecal collections were randomly selected and further examined to determine between and within sample variations of EPG counts as a measure of precision. For this, two random faecal samples were taken from the daily produced faeces by a bird, and the EPG was determined for each of the samples (EPG1 and EPG2). The second faecal sample was analyzed once more to determine a parallel EPG2 count (EPG2a) of the suspended sample. Precision of an EPG count was defined as its relative closeness to the average of two EPG counts using a relative asymmetry index (Index(EPG)). At an age of 11 wk, i.e. 8 wk p.i. the birds were slaughtered and their worm burdens were determined. There were no significant differences between EPG1 and EPG2 (P=0.764) nor between EPG2 and EPG2a (P=0.700), suggesting that the differences between or within the samples were not different from zero. Correlations between EPG counts, as between and within sample coherences, were r=0.85 and r=0.86, respectively. Precision of EPG counts, as measured by Index(EPG), was not influenced by consistency (P=0.870) and total amount of faeces (P=0.088). However, concentration of eggs in faeces (mean EPG) had a significant effect on the precision of the EPG counts (P<0.001). Similar results were also observed for the within sample precision (Index(EPG2)). A segmented regression analysis indicated an abrupt change in the precision of EPG counts as the response to changing egg concentration in the examined faecal samples. The precision of analyses remarkably heightened up to a breakpoint with an EPG count of ≤ 617. A similar breakpoint was also determined for within sample precision (EPG2 ≤ 621). Moderate repeatabilities (R=0.49) for EPG and EPD were estimated in the first week of egg excretion, whereas the estimates were higher (R=0.67-0.84) in the following weeks. Correlations between number of female worms with daily measured EPG and EPD increased to an almost constant level (r ≥ 0.70; P<0.05) in a few days after the nematode excreted eggs and predominantly remained so for the rest of the sampling period. It is concluded that mixing daily total faeces provides samples with random homogenous distribution of H. gallinarum eggs. Precision of the EPG counts increases as the egg concentration in faecal sample increases. Egg excretion of H. gallinarum, quantified either as EPG or EPD, is highly repeatable and closely correlated with the actual worm burden of birds starting as early as in 5 th wk of infection.

Association study in naturally infected helminth layers shows evidence for influence of interferon-gamma gene variants on Ascaridia galli worm burden.

Single nucleotide polymorphisms (SNPs) in the genes for interleukin-4, -13 and interferon-gamma, and 21 additional SNPs which previously had been significantly associated with immune traits in the chicken, were genotyped in white and brown layer hens and analyzed for their association with helminth burden following natural infections. A nucleotide substitution located upstream of the promoter of the interferon-gamma gene was significantly associated with the log transformed number of Ascaridia galli in the brown layer line (genotype CC: 6.4 ± 1.0 worms; genotype CT: 11.7 ± 2.2 worms). Therefore, IFNG seems to be a promising candidate gene for further studies on helminth resistance in the chicken.

Genetic resistance to natural helminth infections in two chicken layer lines.

Groups of Lohmann Brown (LB) and Lohmann Selected Leghorn (LSL) hens were reared under helminth-free conditions and kept afterwards together in a free range system. Mortality rate, body weight development, laying performance and faecal egg counts (FEC) were recorded during a 12 month laying period. At the end of the laying period, 246 LSL and 197 LB hens were necropsied and worms counted following the World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) guidelines. In addition adult Heterakis gallinarum and Ascaridia galli were sexed and measured for length. Significant (P<0.01) differences were observed in mortality rates between LSL and LB animals (12.9 vs. 5.7%). LSL hens showed significantly (P<0.05) higher FEC when compared with LB hens at almost all dates of monitoring. Almost all animals became infected with at least one helminth species. The most prevalent species were H. gallinarum, Capillaria spp. and A. galli. LB hens showed a significantly (P<0.05) higher average number of adult H. gallinarum, Capillaria spp. and tapeworms when compared with LSL animals. However, number of adult A. galli was in tendency lower in these animals. In total, LB had a significantly (P<0.05) higher worm burden than LSL (192.3 vs. 94.3). The estimated heritabilities for total worm burden were 0.23 (SE±0.12) in LSL and 0.75 (SE±0.21) in LB, respectively. The number of all different helminth species were positively correlated. The sex ratio of H. gallinarum and A. galli and the average worm lengths were not significantly (P>0.05) different between the genotypes. There was no significant phenotypic correlation between body weight and worm burden in LSL, whereas it was the case in LB (r=0.17, P<0.05). Based on the estimated heritabilities it is possible to select for helminth resistance in both genotypes.

Effect of extra dietary lysine in Ascaridia galli-infected grower layers.

The hypothesis examined in this study was that extra dietary lysine (Lys) above the recommended standard exerts beneficial effects on the defence reactions, growth performance and nutrient utilization in Ascaridia galli-infected chickens. Therefore, 1-day-old female Lohmann Selected Leghorn chickens were at first fed standard Lys diet (8.5 g Lys/kg DM). At 4 weeks of age, the birds were allocated to groups 1 and 3, both being continued on standard Lys diet, whereas birds in groups 2 and 4 were switched to extra Lys diet (10.5 g Lys/kg DM). Birds in groups 3 and 4 were additionally infected at this age (4 weeks) with 250 embryonated eggs of A. galli. All the birds were slaughtered 7-week-post-infection (p.i.) at an age of 11 weeks. Infected birds on standard Lys diet consumed more feed and reached a similar level of Lys intake as birds on extra Lys diet 7-week-p.i. Utilization of feed, crude protein, and Lys for body weight (BW) gain was adversely affected in infected birds on standard Lys diet (P<0.001). Extra Lys diet reduced the incidence of infection (P<0.05), but worm counts, worm length, faecal egg counts and female worm fecundity were not different between the infected groups (P>0.05). Extra Lys diet led to higher final BW in uninfected and infected birds (P<0.05). It is concluded that extra Lys above the normal recommended standard supports the defence reactions and compensates the negative effect of infection on growth performance and nutrient utilization in A. galli-infected birds.