Kellie Watson

Egg production traits, especially egg number and egg weight, are economically important in chicken breeding programs. However, the genetic basis of these traits in Ethiopian indigenous chicken populations remains unexploited. The study aimed to identify suggestive genomic loci and candidate genes associated with part-period egg number (PPEN; onset of lay to 45 weeks of age) and average egg weight (AEW) in Tilili indigenous chicken population under a selective breeding program. A total of 437 hens were phenotyped and genotyped using low-coverage (0.5x) whole-genome sequencing and imputation against a 30× reference panel. After quality control, 4,846,509 high-quality SNPs were retained for genome-wide association analysis using a linear mixed model. None of the SNPs reached the predefined genome-wide significance threshold ( P < 1 × 10⁻⁸); however, 40 suggestive SNPs for PPEN and 67 suggestive SNPs for AEW ( P < 1 × 10⁻⁵) were identified across 14 chromosomes, consistent with a polygenic genetic architecture for both traits. Candidate genes for PPEN include APOB , AGPATs , CACNA1B , GSTA2 , GSTA3 , GSTA4 , HDAC4 , and MBD3 , whereas HMGA2 , DIAPH3 , SSTR2 , F7 , F10 , and PROZ were associated with AEW. Functional enrichment analysis highlighted pathways related to glutathione metabolism, detoxification, transcriptional regulation, and developmental processes. Overall, the results identify hypothesis-generating genomic regions and candidate genes for egg production traits in Tilili indigenous chickens and provide a basis for further validation in large sample size Tilili chicken population and independent populations.

High genetic variation in African indigenous chicken populations provides opportunities for long-term genetic improvement. This study estimated genetic parameters for economic traits based on data derived from a nucleus flock comprising two generations, derived from 40 sires and 200 dams in a line breeding program through mass selection. Body weight (BW) at different weeks was analyzed for 1370 chickens. Similarly, egg performance was evaluated on 473 hens kept for 44 weeks. Genetic parameters were estimated using a multi-trait animal model based on an average information-restricted maximum likelihood (AI-REML) algorithm in WOMBAT software. Body weight showed significant heritability (p < 0.001), ranging from 0.251 for body weight at 8 weeks of age (BW8) to 0.34 for body weight at 16 weeks of age (BW16), indicating a good genetic improvement potential. Egg production traits had low to moderate heritability (0.08–0.37). Positive genetic correlations among growth traits, particularly BW8 and body weight at 12 weeks of age, BW12 (rG = 0.94), suggest shared genetic influences and the possibility of improving multiple traits simultaneously. The genetic correlation between BW16 and the cumulative egg number varied from low and negative (−0.02) in the first two months to high (0.51) in the cumulative egg number over six months, suggesting that heavier birds lay more eggs over time. Our limited dataset based on two generations and pedigree demonstrates that BW16 with egg production has moderate heritability and strong genetic correlations that can lead to genetic progress toward the development of a dual-purpose breed, and this offers a scientific basis for breeders to develop selection indices to develop a dual-purpose breed for smallholder production systems.

Effective mucosal immunity in the intestine involves a fine balance between tolerance of the microbiome, recognition and elimination of pathogens, and inflammatory tissue injury. The anti-inflammatory cytokine IL10 regulates these processes in the intestines of mice and humans; the anti-inflammatory activity of IL10 is also conserved in birds. To determine the function of IL10 in avian mucosal immunity, we generated germ line modifications of the chicken IL10 locus to abolish or reduce IL10 expression. In vitro analysis of macrophage response to lipopolysaccharide confirmed the loss of IL10 protein expression, the lack of dosage compensation in heterozygotes, and prevention of autocrine inhibition of nitric oxide production in homozygous IL10 knockout macrophages. IL10-deficiency significantly altered the composition of the caecal microbiome, but unlike IL10-deficient mice and humans, IL10-deficient chickens did not exhibit spontaneous colitis. Following experimental challenge with Salmonella enterica serovar Typhimurium or Campylobacter jejuni in IL10-deficient chickens, enhanced clearance of the pathogens was associated with elevated transcription of pro-inflammatory genes and increased infiltration of inflammatory cells into gut mucosa. In IL10-deficient chickens challenged with the parasite Eimeria tenella, pathogen clearance was accelerated but caecal lesions were more severe and weight gain was compromised. Neither the heterozygous IL10 knockout nor a homozygous IL10 enhancer mutation had a major effect on pathogen clearance or inflammation in any of the challenge models. Our findings highlight the intrinsic compromise in mucosal immune response and have important implications for the development of strategies to combat avian and zoonotic pathogens in poultry.

Effective mucosal immunity in the intestine involves a fine balance between tolerance of the microbiome, recognition and elimination of pathogens, and inflammatory tissue injury. The anti-inflammatory cytokine IL10 regulates these processes in the intestines of mice and humans; the anti-inflammatory activity of IL10 is also conserved in birds. To determine the function of IL10 in avian mucosal immunity, we generated germ line modifications of the chicken IL10 locus to abolish or reduce IL10 expression. In vitro analysis of macrophage response to lipopolysaccharide confirmed the loss of IL10 protein expression, the lack of dosage compensation in heterozygotes, and prevention of autocrine inhibition of nitric oxide production in homozygous IL10 knockout macrophages. IL10-deficiency significantly altered the composition of the caecal microbiome, but unlike IL10-deficient mice and humans, IL10-deficient chickens did not exhibit spontaneous colitis. Following experimental challenge with Salmonella enterica serovar Typhimurium or Campylobacter jejuni in IL10-deficient chickens, enhanced clearance of the pathogens was associated with elevated transcription of pro-inflammatory genes and increased infiltration of inflammatory cells into gut mucosa. In IL10-deficient chickens challenged with the parasite Eimeria tenella, pathogen clearance was accelerated but caecal lesions were more severe and weight gain was compromised. Neither the heterozygous IL10 knockout nor a homozygous IL10 enhancer mutation had a major effect on pathogen clearance or inflammation in any of the challenge models. Our findings highlight the intrinsic compromise in mucosal immune response and have important implications for the development of strategies to combat avian and zoonotic pathogens in poultry.

Maximising reproductive success is crucial to animal production systems, particularly in meeting global demands for animal products and improving commercially important traits. However, while social interactions and mating strategies are known to influence reproductive success in wild populations, their consideration in agricultural systems remains limited. Using an interdisciplinary framework that combines concepts from behavioural ecology and quantitative genetics in an animal breeding context, we investigated the role of sperm limitation and polygynous mating strategies (female polyandry, male monopolisation of females and male polygamy) in limiting female reproductive success in farmed Pekin ducks ( Anas platyrthynchos domestica ). We assessed the impact of these behaviours on chick production and quantified their genetic and environmental (co)variance. Our results revealed that the number of dam mates positively influenced chick production in female ducks. However, contrary to our expectation, skew in chick paternity (our measure of male monopolisation) was associated with increased female chick production, challenging the hypothesis that male monopolisation limits the sperm available to females and reduces their reproductive success. We found no evidence that male polygamy led to decreased female chick production. Genetic analysis revealed that female mate number and reproductive skew exhibit genetic variance, providing opportunities for targeted selection to enhance chick production. However, there was a negative genetic association between female polyandry and skew in chick paternity, suggesting a trade-off between these traits that would need to be considered in future selection programmes. Our findings highlight how concepts from behavioural ecology can be incorporated into breeding programmes, providing new opportunities to develop effective and sustainable breeding strategies.

Chicken is a valuable model for understanding fundamental biology and vertebrate evolution, as well as a major source of nutrient-dense and lean-protein-enriched food globally. Although it is the first non-mammalian amniote genome to be sequenced, the chicken genome still lacks a systematic characterization of functional variation. Here, through integrating bulk RNA-Seq of 7,015 samples, single-cell RNA-Seq of 127,598 cells and 2,869 whole-genome sequences, we presented an atlas of regulatory variants across 28 chicken tissues and 43 cell types, including millions of regulatory effects on primary expression (protein-coding genes, lncRNA and exon) and post-transcriptional modifications (alternative splicing and 3’ untranslated region alternative polyadenylation). We highlighted distinct molecular mechanisms underlying these regulatory variants and their context dependence and utility in interpreting genome-wide associations of 108 chicken complex traits. Finally, we illustrated lineage-specific features of gene regulation between chickens and mammals, and demonstrated how this resource can assist with cross-species gene mapping of complex traits.

Chicken is a valuable model for understanding fundamental biology, vertebrate evolution and diseases, as well as a major source of nutrient-dense and lean-protein-enriched food globally. Although it is the first non-mammalian amniote genome to be sequenced, the chicken genome still lacks a systematic characterization of functional impacts of genetic variants. Here, through integrating 7,015 RNA-Seq and 2,869 whole-genome sequence data, the Chicken Genotype- Tissue Expression (ChickenGTEx) project presents the pilot reference of regulatory variants in 28 chicken tissue transcriptomes, including millions of regulatory effects on primary expression (including protein-coding genes, lncRNA and exon) and post-transcriptional modifications (alternative splicing and 3’ untranslated region alternative polyadenylation). We explored the tissue-sharing and context-specificity of these regulatory variants, their underlying molecular mechanisms of action, and their utility in interpreting adaptation and genome-wide associations of 108 chicken complex traits. Finally, we illustrated shared and lineage-specific features of gene regulation between chickens and mammals, and demonstrated how the ChickenGTEx resource can further assist with translating genetic findings across species. One-Sentence Summary: The ChickenGTEx provides a multi-tissue reference of regulatory variants for chicken genetics and genomics, functional genomics, precision breeding, veterinary medicine, vertebrate evolution and even human biomedicine.

Coccidiosis is a major contributor to losses in poultry production. With emerging constraints on the use of in-feed prophylactic anticoccidial drugs and the relatively high costs of effective vaccines, there are commercial incentives to breed chickens with greater resistance to this important production disease. To identify phenotypic biomarkers that are associated with the production impacts of coccidiosis, and to assess their covariance and heritability, 942 Cobb500 commercial broilers were subjected to a defined challenge with Eimeria tenella (Houghton). Three traits were measured: weight gain (WG) during the period of infection, caecal lesion score (CLS) post mortem, and the level of a serum biomarker of intestinal inflammation, i.e. circulating interleukin 10 (IL-10), measured at the height of the infection. Phenotypic analysis of the challenged chicken cohort revealed a significant positive correlation between CLS and IL-10, with significant negative correlations of both these traits with WG. Eigenanalysis of phenotypic covariances between measured traits revealed three distinct eigenvectors. Trait weightings of the first eigenvector, (EV1, eigenvalue = 59%), were biologically interpreted as representing a response of birds that were susceptible to infection, with low WG, high CLS and high IL-10. Similarly, the second eigenvector represented infection resilience/resistance (EV2, 22%; high WG, low CLS and high IL-10), and the third eigenvector tolerance (EV3, 19%; high WG, high CLS and low IL-10), respectively. Genome-wide association studies (GWAS) identified two SNPs that were associated with WG at the suggestive level. Eigenanalysis separated the phenotypic impact of a defined challenge with E. tenella on WG, caecal inflammation/pathology, and production of IL-10 into three major eigenvectors, indicating that the susceptibility-resistance axis is not a single continuous quantitative trait. The SNPs identified by the GWAS for body weight were located in close proximity to two genes that are involved in innate immunity (FAM96B and RRAD).