Short- and long-term consequences of stocking density during rearing on the immune system and welfare of laying hens.

Already during early life, chickens need to cope with chronic stressors that can impair their health and welfare, with stocking density being one of the most influential factors. Nevertheless, there is a gap in research on the influence of stocking density on laying hens during rearing and in the subsequent laying period. This study therefore investigated how stocking density during rearing affects the immune system and welfare of pullets, and whether effects are persistent later in life. Pullets were reared at either low (13 birds/m2) or high (23 birds/m2) stocking densities but in identical group sizes from wk 7 to 17. Afterward, hens were kept at the same stocking density (2.4 birds/m2) until wk 28. Blood and tissue samples (spleen and cecal tonsils) were collected at the end of the rearing period and in the laying period. The parameters evaluated encompassed number and distribution of leukocytes and lymphocyte subsets in blood and lymphatic tissue, lymphocyte functionality, plasma corticosterone concentrations as well as behavior and physical appearance of hens. At the end of rearing, pullets kept under high stocking density had lower numbers of T lymphocytes, especially γδ T cells in blood, spleen, and cecal tonsils and displayed a higher heterophil to lymphocyte ratio. These effects are mostly persistent during the laying period, although stocking density was identical at this time. Furthermore, birds from the high stocking density group showed less active behavior, more pecking behavior and worse physical appearance throughout both examination periods. In conclusion, stocking density during rearing affects pullets' immune system and behavior not only in the rearing, but also subsequently in the laying period, indicating a strong correlation between health and welfare during rearing and the laying period.

Study of the antibiotic residues in poultry meat in some of the EU countries and selection of the best compositions of lactic acid bacteria and essential oils against Salmonella enterica.

In this study, the presence of antibiotics (ANB) residues was evaluated in poultry meat purchased from German and Lithuanian markets. In addition, the antimicrobial activity of 13 lactic acid bacteria (LAB) strains, 2 essential oils (EO) (Thymus vulgaris and Origanum vulgare L.), and their compositions were tested for the purpose of inhibiting antibiotic-resistant Salmonella spp. ANB residues were found in 3 out of the 20 analyzed poultry meat samples: sample no. 8 contained enrofloxacin (0.46 µg/kg), sample no. 14 contained both enrofloxacin and doxycycline (0.05 and 16.8 µg/kg, respectively), and sample no. 18 contained enrofloxacin (2.06 µg/kg). The maximum residue limits (MRLs) for the sum of enrofloxacin and ciprofloxacin and for doxycycline in the poultry muscle are 100 µg/kg. Finally, none of the tested poultry meat samples exceeded the suggested MRLs; however, the issue of ANB residues still requires monitoring of the poultry industry in Germany, Poland, and Lithuania, despite the currently established low ANB concentrations. These findings can be explained by the increased use of alternatives to ANB in the poultry industry. Our results showed that an effective alternative to ANB, which can help to reduce the occurrence of antibiotic-resistant salmonella, is a composition containing 1.0% of thyme EO and the following LAB strains: Lactobacillus plantrum LUHS122, Enteroccocus pseudoavium LUHS242, Lactobacillus casei LUHS210, Lactobacillus paracasei LUHS244, Lactobacillus plantarum LUHS135, Lactobacillus coryniformins LUHS71, and Lactobacillus uvarum LUHS245, which can be recommended for poultry industry as components of feed or for the treatment of surfaces, to control the contamination with Salmonella strains. However, it should be mentioned that most of the tested LAB strains were inhibited by thyme EO at the concentrations of 0.5 and 1.0%, except for LUHS122, LUHS210, and LUHS245. Finally, it can be noted that the agents responsible for the inhibitory effect on Salmonella are not the viable LAB strains but rather their metabolites, and further studies are needed to identify which metabolites are the most important.

Impact of Housing Environment on the Immune System in Chickens: A Review.

During their lifespan, chickens are confronted with a wide range of acute and chronic stressors in their housing environment that may threaten their welfare and health by modulating the immune system. Especially chronic stressful conditions can exceed the individual's allostatic load, with negative consequences for immunity. A fully functional immune system is mandatory for health and welfare and, consequently, also for high productivity and safe animal products. This review provides a comprehensive overview of the impact of housing form, light regime as well as aerial ammonia and hydrogen sulfide concentrations on the immune system in chickens. Certain housing conditions are clearly associated with immunological alterations which potentially impair the success of vaccinations or affect disease susceptibility. Such poor conditions counteract sustainable poultry production. This review also outlines current knowledge gaps and provides recommendations for future research.

Resistance and tolerance to mixed nematode infections in relation to performance level in laying hens.

Modern chickens have been genetically developed to perform high under optimal conditions. We hypothesized that high-performance is associated with a higher sensitivity to environmental challenges in laying hens. By using nematode infections as an environmental stressor, we assessed performance-level associated host responses in a high (i.e. Lohmann Brown Plus, LB) and in a lower performing, a so-called dual-purpose chicken genotype (i.e. Lohmann Dual, LD). The hens were infected with 1000 eggs of Ascaridia galli and Heterakis gallinarum at 24 weeks of age. Hen performance parameters, humoral immune responses in plasma and egg yolks and worm burdens were assessed at several occasions over a period of 18 weeks post infection (wpi). While infections had no significant effect on feed intake (P = 0.130) and body weight in both genotypes (P = 0.392), feed conversion efficiency was negatively affected by infections (P = 0.017). Infections reduced both laying rate and egg weight and thereby per capita egg mass in both genotypes (P < 0.05). While laying rate in infected LB hens decreased significantly (P < 0.05) in the early infection period (i.e. by 3 wpi), the decrease in LD hens appeared much later (i.e. by 14 wpi). Worm burdens resulting from the experimental infection were not different between the genotypes for both worm species (P > 0.05), whereas LB hens were more susceptible (P < 0.05) to re-infections than LD hens. Changes in humoral immune responses (i.e. ascarid-specific IgY antibodies in plasma and egg yolks) of the two genotypes over time reflected closely the corresponding changes in larval counts of the hens, descending from both experimental and subsequent natural infections in both genotypes. Infections caused a shift in egg size classes, leading to smaller frequency of larger eggs in both genotypes. Infections reduced egg weight (P = 0.018) and led to a reduced fat content in the egg yolks (P = 0.045). The proportion of poly-unsaturated fatty acids (PUFA), especially n-6-PUFA, was also lower in egg yolks of the infected hens (P = 0.032). We conclude that tolerance to nematode infections in laying hens is dependent on host-performance level. The impairment in host tolerance was both genotype and time dependent, likely due to differences in genetic programming for production peak and persistency of the two genotypes. The two genotypes exhibited similar levels of resistance after a fully controlled experimental infection, but the high performing hens were more susceptible to subsequent natural infections. Infections negatively affected economically important egg-quality traits, including egg weight, fat content and fatty acid profiles in egg yolks.

A genome-wide association study in a large F2-cross of laying hens reveals novel genomic regions associated with feather pecking and aggressive pecking behavior.

BACKGROUND: Feather pecking and aggressive pecking in laying hens are serious economic and welfare issues. In spite of extensive research on feather pecking during the last decades, the motivation for this behavior is still not clear. A small to moderate heritability has frequently been reported for these traits. Recently, we identified several single-nucleotide polymorphisms (SNPs) associated with feather pecking by mapping selection signatures in two divergent feather pecking lines. Here, we performed a genome-wide association analysis (GWAS) for feather pecking and aggressive pecking behavior, then combined the results with those from the recent selection signature experiment, and linked them to those obtained from a differential gene expression study. METHODS: A large F2 cross of 960 F2 hens was generated using the divergent lines as founders. Hens were phenotyped for feather pecks delivered (FPD), aggressive pecks delivered (APD), and aggressive pecks received (APR). Individuals were genotyped with the Illumina 60K chicken Infinium iSelect chip. After data filtering, 29,376 SNPs remained for analyses. Single-marker GWAS was performed using a Poisson model. The results were combined with those from the selection signature experiment using Fisher's combined probability test. RESULTS: Numerous significant SNPs were identified for all traits but with low false discovery rates. Nearly all significant SNPs were located in clusters that spanned a maximum of 3 Mb and included at least two significant SNPs. For FPD, four clusters were identified, which increased to 13 based on the meta-analysis (FPDmeta). Seven clusters were identified for APD and three for APR. Eight genes (of the 750 investigated genes located in the FPDmeta clusters) were significantly differentially-expressed in the brain of hens from both lines. One gene, SLC12A9, and the positional candidate gene for APD, GNG2, may be linked to the monomanine signaling pathway, which is involved in feather pecking and aggressive behavior. CONCLUSIONS: Combining the results from the GWAS with those of the selection signature experiment substantially increased the statistical power. The behavioral traits were controlled by many genes with small effects and no single SNP had effects large enough to justify its use in marker-assisted selection.

Genetic parameters and signatures of selection in two divergent laying hen lines selected for feather pecking behaviour.

BACKGROUND: Feather pecking (FP) in laying hens is a well-known and multi-factorial behaviour with a genetic background. In a selection experiment, two lines were developed for 11 generations for high (HFP) and low (LFP) feather pecking, respectively. Starting with the second generation of selection, there was a constant difference in mean number of FP bouts between both lines. We used the data from this experiment to perform a quantitative genetic analysis and to map selection signatures. METHODS: Pedigree and phenotypic data were available for the last six generations of both lines. Univariate quantitative genetic analyses were conducted using mixed linear and generalized mixed linear models assuming a Poisson distribution. Selection signatures were mapped using 33,228 single nucleotide polymorphisms (SNPs) genotyped on 41 HFP and 34 LFP individuals of generation 11. For each SNP, we estimated Wright's fixation index (FST). We tested the null hypothesis that FST is driven purely by genetic drift against the alternative hypothesis that it is driven by genetic drift and selection. RESULTS: The mixed linear model failed to analyze the LFP data because of the large number of 0s in the observation vector. The Poisson model fitted the data well and revealed a small but continuous genetic trend in both lines. Most of the 17 genome-wide significant SNPs were located on chromosomes 3 and 4. Thirteen clusters with at least two significant SNPs within an interval of 3 Mb maximum were identified. Two clusters were mapped on chromosomes 3, 4, 8 and 19. Of the 17 genome-wide significant SNPs, 12 were located within the identified clusters. This indicates a non-random distribution of significant SNPs and points to the presence of selection sweeps. CONCLUSIONS: Data on FP should be analysed using generalised linear mixed models assuming a Poisson distribution, especially if the number of FP bouts is small and the distribution is heavily peaked at 0. The FST-based approach was suitable to map selection signatures that need to be confirmed by linkage or association mapping.

Quantitative genetic analysis of traits related to fear and feather pecking in laying hens.

Feather pecking is a well known problem in flocks of laying hens. It is partially controlled by genetics. Fear is frequently reported to be related with feather pecking. The present study reports the result from a quantitative genetic analysis of feather pecking and three fear test traits in laying hens. Fear was recorded by the tonic immobility test, the open field activity and the emergence box test. These were recorded at a juvenile and adult age of the hens. The heritability of feather pecking was 0.16, and in the range between 0.07 and 0.14 for the fear test traits. Genetic correlations between fear measured in the juvenile and in the adult age point to different but correlated traits. Tonic immobility measured early in life was moderately correlated with feather pecking and might be used as a breeding criterion to reduce feather pecking.