British Poultry Science

Wilson, P. W.; Dunn, I. C.; Mccormack, H. A.

doi: 10.1080/00071668.2022.2119835pmid: 36196860

1. Genetic selection for bone quality can improve this, as it is heritable. A method was established using digital X-ray which took around 40 s in total and gave an image that allowed quantification of bone density from many appendicular bones. 2. The tibiotarsus measurement of bone density on the live hen across the different experiments had correlations with post-mortem whole bone radiographic density from 0.62 to 0.7, similar to that between density and material properties for example. Differences between groups of hens, where calcium and phosphorus in the diet were manipulated, were detected within 3 weeks of treatment using live hen measurement (P < 0.001, n = 24). 3. In a gage analysis, ‘hen’ explained more than 86% of the variance, demonstrating the ability to observe clear differences between hens. The effect of different operators’ analysis on the contribution to variance was very low as was the repeated measurement of the same hen. 4. The measurement of bone density on the live hen described in this paper represented major progress to a usable method for genetic selection to improve bone strength in laying hens. The method has the potential to reduce the number of animals needed to test nutritional and management interventions to improve bone health.

Lin, Z. T.; Chen, G. H.; Peng, X.; Zhang, Z. H.; Li, T.; Lin, H. X.; Liang, S. S.; Zheng, Y. B.; Yao, Z. P.; Luo, W.

doi: 10.1080/00071668.2022.2094217pmid: 35759289

1. Searching for molecular markers related to growth and carcase traits plays a critical role in improvement of the production performance of broilers. Previous studies found that transmembrane protein 182 (TMEM182) inhibits skeletal muscle development, growth, and regeneration, implying that the TMEM182 gene plays an important role during the development process of skeletal muscle. 2. A novel 2-bp indel in intron 1 of TMEM182 was detected in a yellow chicken population derived from the cross of White Recessive Rock chickens with Xinghua chickens, and three genotypes II (inserted homozygote), ID (inserted and deleted heterozygote) and DD (deleted homozygote) were observed. Association analyses indicated that the indel was significantly associated with the body weight, muscle fibre area, breast muscle weight and wing weight in the F2 population. 3. The expression of TMEM182 in leg muscle of chickens with II genotype was higher than that with DD genotype, with the 2-bp indel located in one of the putative PAX4 binding sites. Further research through luciferase assays revealed that the PAX4 could bind to the putative binding site and increase the TMEM182 transcription, with the 2-bp deletion disrupting the binding of PAX4. 4. The present study provides evidence for the association of the novel 2-bp indel in intron 1 of TMEM182 with the growth and carcase traits of chickens. This 2-bp indel could be used as a genetic marker in broiler breeding.

Yu, S.; Wang, G.; Liao, J.; Shen, X.; Chen, J.

doi: 10.1080/00071668.2022.2113506pmid: 35979716

1. Long non-coding RNAs (lncRNAs) play important roles in various physiological functions. However, the mechanisms underlying the regulation of lncRNAs in melanogenesis remain unclear. To determine the molecular mechanisms involved in skin melanogenesis, the present study depicted the expression profiles of lncRNAs and messenger RNAs (mRNAs) in black- (B group) and white- (W group) skinned chickens using RNA sequencing. 2. In total, 373 differentially expressed lncRNAs (DELs; 203 up-regulated and 170 down-regulated) and 253 differentially expressed genes (DEGs; 152 up-regulated and 101 down-regulated) were identified between the B and W groups. A total of eight known melanogenesis-related genes were identified (KIT, TYRP1, DCT (TYRP2), SLC45A2, OCA2, EDNRB2, TRPM1 and RAB38). 3. Functional annotation of the co-expressed DEGs and DELs was performed using Gene Ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analyses. The co-expressed DEGs were mainly involved in melanogenesis and the co-expressed genes of 117 and 108 DELs were significantly enriched in the melanogenesis and tyrosine metabolism pathways, respectively. 4. The DEL-DEG interaction network revealed that three lncRNAs (XR_003072387.1, XR_003075112.1, and XR_003077033.1) and DCT genes may have key roles in regulating melanogenesis in chicken skin. This data provides the groundwork for studying the lncRNA regulatory mechanisms of skin melanogenesis and suggested a new perspective on the modulation of melanogenesis in chicken skin based on a lncRNA-mRNA causal regulatory network.

Li, J.; Ma, Y.; Zhang, L.; Cai, C.; Guo, Y.; Zhang, Z.; Li, D.; Tian, Y.; Kang, X.; Han, R.; Jiang, R.

doi: 10.1080/00071668.2022.2121640pmid: 36102935

1. This study explored the tissue metabolic status and the relationship with inflammation in valgus-valgus deformity (VVD) broilers with increasing age. 2. Tissue and blood from VVD and healthy broilers were collected at two, four and five weeks old. A fully automated biochemical analyser, real-time PCR, HE staining and enzyme-linked immunosorbent assay were used to detect tissue metabolic indexes, mRNA levels of inflammation and apoptosis cytokines in immune organs, histological changes and serum inflammation and immune-related protein contents in VVD broilers. 3. The results showed that VVD increased the levels of total protein, albumin, alanine aminotransferase at five weeks of age, aspartate aminotransferase, urea and creatine kinase in blood at two weeks of age. It upregulated the gene expression of inflammatory factors IL-1β, IL-6, IL-8, TNF-α, NF-κB and TGF-β and apoptotic factors FAS, Bcl-2, caspase-3 and 9 in immune organs; increased levels of serum proteins TNF-α, IL-1β and IL-6 and decreased levels of serum immunoglobulins IgY and CD3+. 4. In addition, with increasing age, IL-10 gene expression gradually increased in the BF and decreased in the spleen. 5. In conclusion, VVD broilers have disorders of liver and kidney metabolism, inflammation and apoptosis of immune organs and increased levels of serum inflammatory factor proteins.

Guo, Y.; Zhang, J.; Li, X.; Wu, J.; Han, J.; Yang, G.; Zhang, L.

doi: 10.1080/00071668.2022.2122025pmid: 36083210

1. Ammonia is one of major air pollutants in intensive poultry houses, where it causes immunosuppression in broilers. Although previous studies have focused on a particular organ, data on multiple organs have not been reported. 2. In the following work, broilers were exposed to environmental ammonia (0, 10, 20, and 40 mg/m3 from 1–21 d old; and 0, 15, 30, and 60 mg/m3 from 22–42 d old). 3. Ammonia exposure reduced bird spleen index at 42 d and thymus index at 14, 28, 35 and 42 d, meaning that ammonia caused immunosuppression in birds. Moreover, high ammonia exposure down-regulated the expression of toll-like receptor 4 (TLR4) in lung tissue at 21 d, as well as TLR4 in lung and tracheal mucosa at 42 d when analysed using qRT-PCR. It increased SIgA in saliva at 42 d when analysed by ELISA. Ammonia increased interleukin-6 (IL-6), IL-1β, interferon-α (IFN-α), and IFN-γ in serum at 28 d from the ELISA assay, which indicated that all of these factors took part in ammonia-immunosuppression in birds. 4. Three antioxidants (CAT, SOD, T-AOC) decreased, and one oxidant MDA increased after ammonia exposure in the liver and blood, which indicated that ammonia caused oxidative stress via the imbalance of antioxidants/oxidants in birds. 5. Correlation analysis showed that TLR4 and TLR15 in the tracheal mucosa were significantly positively related to IFN-γ and negatively related to IL-6. TLR2 in the lung was significantly positively related to IL-1β, and TLR2 in bird tracheal mucosa was negatively related to IL-6 in serum. 6. The results suggested that oxidative stress mediated immunosuppression caused by ammonia gas via antioxidant/oxidant imbalance in broilers.

Takahashi, T.; Suzuki, N.; Ishii, R.; Toyoda, S.; Shibata, M.; Azuma, Y.; Kurose, Y.

doi: 10.1080/00071668.2022.2126933pmid: 36129068

1. This study assessed 1) the effects of Paracoccus carotinifaciens supplementation containing high astaxanthin levels on egg production performance and quality, 2) dynamics of carotenoids levels in the egg yolk and 3) taste of astaxanthin-rich egg yolk. 2. Laying hens were fed diets containing different levels of P. carotinifaciens-derived astaxanthin (ASX; 0, 2, 4, 8, or 16 ppm) for 28 d (experiment 1) or a diet containing 16 ppm astaxanthin for 28 d followed by a 0 ppm astaxanthin diet for 28 days (experiment 2). 3. Production performance, egg quality and egg yolk carotenoid levels were examined in experiment 1 (Ex1) and the dynamics of egg yolk carotenoid levels and egg yolk taste in experiment 2 (Ex2). 4. ASX supplementation did not affect production performance or egg quality. ASX levels in the egg yolk became saturated after seven days of 16 ppm supplementation and decreased to less than one-tenth of the saturated levels seven days after supplementation cessation. Supplementation with 16 ppm ASX for 28 d did not affect egg yolk taste. 5. Supplementation resulted in the production of ASX-rich eggs for a brief period without affecting production performance, egg quality or taste. Understanding the time taken for the incorporation of ASX into egg yolks is beneficial for value-added egg production and may help in minimising supplementation costs.

Gumus, D.; Kizil, M.

doi: 10.1080/00071668.2022.2126932pmid: 36129064

1. The objective of the present study was to examine the effect of propolis extract on reducing the formation of carcinogenic/mutagenic heterocyclic aromatic amines (HAAs), thereby minimising dietary exposure in human consumers. 2. Chicken thigh meat samples were marinated with various concentrations (0%, 0.25%, 0.5% and 1%) of propolis extract, and cooked in a pan at 150°C or 200°C. Proximate composition, pH, lipid oxidation, creatine, creatinine content and twelve HAA levels of samples were analysed. 3. Varying levels of IQx (≤35.44 ng/g), MeIQx (≤0.58 ng/g), MeIQ (≤1.60 ng/g), 7,8-DiMeIQx (≤0.83 ng/g), 4,8-DiMeIQx (≤0.75 ng/g), Harman (≤5.54 ng/g), Trp-P-2 (≤1.77 ng/g), PhIP (≤1.61 ng/g) and AαC (≤0.93 ng/g) were quantified in control samples. Total HAA levels ranged between 2.83 and 47.26 ng/g across all samples. Propolis extract decreased the levels of total HAAs by 41.2–89.4% and 49.4–91.4% at 150°C and 200°C, respectively. 4. The results demonstrated that propolis extract marination might be an effective strategy to reduce the dietary exposure of HAAs via mitigating their formation in chicken thigh meat.

Dishan, A.; Hizlisoy, H.; Barel, M.; Disli, H. B.; Gungor, C.; Ertas Onmaz, N.; Gonulalan, Z.; Al, S.; Yildirim, Y.

doi: 10.1080/00071668.2022.2116697pmid: 36102939

1. The Shiga toxin-producing Escherichia coli (STEC) is a hazardous zoonotic agent for chicken meat consumers. This study determined the serogroups and evaluated the virulence genes, antibiotic resistance, biofilm-forming profiles and genetic relationships of STEC isolates in chicken meat. 2. A total of 100 samples belonging to dressed-whole chicken and different parts of the chicken (wing, breast, thigh, drumstick) were collected between September and November 2019 from different retail markets in Kayseri, Türkiye. 3. Phenotypic (identification, disc diffusion test, Congo red agar and microtitre plate tests) and molecular tests (identification, serogrouping, virulence factors, biofilm, antibiotic susceptibility, 16S rRNA sequencing and enterobacterial repetitive intergenic consensus-PCR for typing of the isolates) were carried out. 4. E. coli was isolated from 35% of the samples and 35% of the samples harboured at least one STEC. Among 35 STEC isolates, 3 (8.5%), 6 (17.1%), 2 (5.7%) and 3 (8.5%) were found to be positive for fliCH2, fliCH8, fliCH11, fliCH19 genes, respectively. Out of 35 STEC positive isolates, 4 (11.4%) were identified as E. coli O157, from which 2 (5.7%) were E. coli O157:H7. E. coli O157 was detected in two (10%), one (5%), one (5%) of the thigh, drumstick and whole chicken samples, respectively. 5. Biofilm-forming ability was reported in 33 (94.2%) of 35 E. coli isolates, whilst the biofilm-associated genes detected among 35 STEC isolates included csgA (88.5%), fimH (88.5%), bcsA (85.7%), agn43 (14.2%) and papC (8.5%). The STEC strains showed resistance against ampicillin (88.5%) and erythromycin (88.5%), followed by tetracycline (74.2%) and gentamicin (25.7%). However, the distribution of isolates harbouring bla CMY, ere(A), tet(A) and aac(3)-IV antibiotic resistance genes was found to be 17.1%, 11.4%, 85.7% and 5.7%, respectively. 6. ERIC-PCR showed that E. coli strains obtained from different parts and whole of chicken samples had genetic diversities. ERIC-PCR patterns grouped strains of 35 STEC into eight clusters designated A-H, with 73% similarity. Proper hygiene measures and staff training are essential for public health during poultry processing and in retail stores to control STEC.

Wang, S.; Zhao, J.; Wang, L.; Zhang, T.; Zeng, W.; Lu, H.

doi: 10.1080/00071668.2022.2121639pmid: 36069737

1. Methyltransferase-like 21C (METTL21C) and insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) play important roles in the proliferation of chicken myoblasts. However, it remains unclear whether there is protein-protein interaction between METTL21C and IGF2BP1 to regulate proliferation of chicken myoblasts. 2. In this study, the Igf2bp1 gene was amplified from cDNA of liver tissue of Lueyang black-bone chicken to construct the overexpression vector HA-Igf2bp1. The HA-Igf2bp1 and Flag-Mettl21c vectors were individually transfected and co-transfected into HEK293T, respectively. Co-immunoprecipitation (Co-IP) assay indicated a protein-protein interaction between METTL21C and IGF2BP1. 3. Using the Western blotting and LC-MS/MS, it was found that METTL21C could mediate the lysine methylation modification of IGF2BP1. Furthermore, the His-tagged overexpression vector HA-Igf2bp1-His was constructed, transfected and co-transfected with Flag-Mettl21c into HEK293T. His-tagged IGF2BP1 was purified by nickel ion affinity chromatography. Western blotting revealed that IGF2BP1 was successfully purified, and the trimethylation modification level of co-transfection group was significantly elevated compared with the single-transfection Igf2bp1 group. 4. Mettl21c and Igf2bp1 overexpression vectors were transfected and co-transfected into primary chicken myoblasts, respectively. The results of 5-ethynyl-2’-deoxyuridine assay and the expression level of Pax7 and MyoD indicated that overexpression of Igf2bp1 alone inhibited the chicken myoblast proliferation, whereas co-expression of Mettl21c and Igf2bp1 eliminated the inhibitory effects of Igf2bp1, thereby favouring cell proliferation and differentiation. 5. The results, for the first time, revealed that METTL21C mediated the lysine trimethylation modification of IGF2BP1 to regulate the proliferation of chicken myoblasts, which provided a new insight into in-depth analysis of the molecular mechanism of METTL21C methylation involved in regulating the growth and development of skeletal muscle in Lueyang black-bone chicken.

Alan, E.; Liman, N.; Sağsöz, H.

doi: 10.1080/00071668.2022.2121912pmid: 36083170

1. The epidermal growth factor (EGF) family plays an important role in the development, differentiation, migration and apoptosis of cells, as well as in wound healing, which are all essential to the viability of multicellular organisms. The avian spleen is a principal organ of systemic immunity and its importance in disease resistance is presumably accentuated by the scarcity of avian lymph nodes. 2. The aim of this study was to determine whether EGF receptors (ErbB1-4) and their ligands (EGF, AREG and NRG) are expressed in the structural components of the quail spleen during the post-hatch period. At each selected age, from 1 d to 7, 14, 21 and 60 d, 10 quails were euthanised under ether anaesthesia and their spleens were fixed in a 10% formaldehyde-alcohol solution. Following routine histological processing, the streptavidin-biotin-peroxidase method was used for immunohistochemical examination. 3. Strong cytoplasmic immunoreactions for ErbB2, ErbB4 and NRG were observed in the ellipsoid associated cells (EAC) of the quail spleen throughout the post-hatch period. This immunoreactivity in the EAC increased after the 7th d post-hatch. ErbB1 and ErbB3 immunoreactions were relatively similar and weak in all components of the spleen during the post-hatch period. Some immune cells of the peri-arterial lymphatic sheath (PALS) and peri-ellipsoidal lymphatic sheath (PELS) showed positive immunoreactivity for the ErbB receptors and their ligands. In the vascular smooth muscle cells, immunoreactivity for ErbB2 was stronger than that for the other ErbB receptors and their ligands. 4. The data showed that ErbB receptors and their ligands (EGF, AREG and NRG) are expressed by different structural components of the quail spleen during the post-hatch period.