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Intestinal microbiome profiles in broiler chickens raised without antibiotics exhibit altered microbiome dynamics relative to conventionally raised chickens.

The present study was undertaken to profile and compare the cecal microbial communities in conventionally (CONV) grown and raised without antibiotics (RWA) broiler chickens. Three hundred chickens were collected from five CONV and five RWA chicken farms on days 10, 24, and 35 of age. Microbial genomic DNA was extracted from cecal contents, and the V4-V5 hypervariable regions of the 16S rRNA gene were amplified and sequenced. Analysis of 16S rRNA sequence data indicated significant differences in the cecal microbial diversity and composition between CONV and RWA chickens on days 10, 24, and 35 days of age. On days 10 and 24, CONV chickens had higher richness and diversity of the cecal microbiome relative to RWA chickens. However, on day 35, this pattern reversed such that RWA chickens had higher richness and diversity of the cecal microbiome than the CONV groups. On days 10 and 24, the microbiomes of both CONV and RWA chickens were dominated by members of the phylum Firmicutes. On day 35, while Firmicutes remained dominant in the RWA chickens, the microbiome of CONV chickens exhibited am abundance of Bacteroidetes. The cecal microbiome of CONV chickens was enriched with the genus Faecalibacterium, Pseudoflavonifractor, unclassified Clostridium_IV, Bacteroides, Alistipes, and Butyricimonas, whereas the cecal microbiome of RWA chickens was enriched with genus Anaerofilum, Butyricicoccu, Clostridium_XlVb and unclassified Lachnospiraceae. Overall, the cecal microbiome richness, diversity, and composition were greatly influenced by the management program applied in these farms. These findings provide a foundation for further research on tailoring feed formulation or developing a consortium to modify the gut microbiome composition of RWA chickens.

Animals

GCRP: Integrated Global Chicken Reference Panel from 11,951 Chicken Genomes.

Chickens are a crucial source of protein for humans and a popular model animal for bird research. Despite the emergence of imputation as a reliable genotyping strategy for large populations, the lack of a high-quality chicken reference panel has hindered progress in chicken genome research. To address this, here we introduce the first phase of the 100K Global Chicken Reference Panel (100K GCRP). Currently, two panels are available: a comprehensive mix panel (CMP) for domestication diversity research and a commercial breed panel (CBP) for breeding broilers specifically. Evaluation of genotype imputation quality showed that CMP had the highest imputation accuracy compared to imputation using existing chicken panels in Animal-SNPAtlas and Animal Genotype Imputation Database (AGIDB), whereas CBP performed stably in the imputation of commercial populations. Additionally, we found that genome-wide association studies using GCRP-imputed data, whether on simulated or real phenotypes, exhibited greater statistical power. In conclusion, our study indicates that the GCRP effectively fills the gap in high-quality reference panels for chickens, providing an effective imputation platform for future genetic and breeding research. The project includes 11,951 samples and provides services for various applications on its website at http://farmrefpanel.com/GCRP/#/.

Animals

Comparative Analysis of Volatile Compounds, Amino Acids, Fatty Acids, and Lipidomic Profiles in Thigh Muscles of Commercial Arbor Acres (AA) Broilers and Indigenous Chengkou and Langshan Chickens.

Flavor-related compounds and nutritional components of chicken meat vary among different breeds, but comprehensive comparisons of these characteristics between commercial and indigenous chickens remain insufficiently characterized. In this study, three chicken breeds (Arbor Acres, Chengkou, and Langshan) were slaughtered at their respective market ages, and the volatile flavor compounds, amino acids, fatty acids, and lipidomic profiles of thigh muscle were analyzed to investigate breed-associated differences in flavor-related and nutritional characteristics. Langshan chickens exhibited the highest total volatile compound content and also had the highest total amino acid levels, with significantly higher contents of umami and sweet amino acids. In addition, both indigenous breeds showed higher levels of arachidonic acid (C20:4n6) than Arbor Acres broilers, while Chengkou chickens had the highest content of docosahexaenoic acid (DHA, C22:6n3). Lipidomic analysis identified 787 lipids, with glycerophospholipids and sphingolipids as the predominant classes. Differential lipid analysis revealed that Langshan chickens had 38 upregulated lipids compared with Arbor Acres chickens, while Chengkou chickens exhibited 258 differential lipids relative to Arbor Acres chickens. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis indicated that these differential lipids were mainly associated with glycerolipid, sphingolipid, and glycerophospholipid metabolism. Correlation analysis further revealed significant associations between specific lipids and flavor-related compounds, amino acids, and fatty acids, suggesting their potential roles in breed-associated differences. Overall, this study demonstrates that indigenous chicken breeds possess distinct flavor-related and nutritional profiles compared with commercial Arbor Acres broilers and provides valuable insights into breed-associated differences in chicken meat characteristics.

amino acids

Low-pass whole-genome sequencing reveals genomic diversity and ecotype-specific adaptation in indigenous Tigrayan chickens.

Indigenous chickens play a critical role in food security and climate resilience in smallholder systems, yet their genomic diversity and adaptive potential remain insufficiently characterised. This study employed low-pass whole-genome sequencing (LP-WGS; 0.2-1.99×) to investigate genomic diversity, population structure, inbreeding and candidate environment-associated genomic variation in 33 chickens from highland, midland, and lowland agroecologies in the Tigray region of northern Ethiopia. After imputation and stringent filtering, 23.4 million high-confidence SNPs were retained, including ~ 17% novel variants, indicating substantial uncharacterised genetic diversity in these populations. SNP density (13.8 ± 8.6 SNPs/kb) was comparable to values reported from high-coverage Ethiopian chicken datasets, demonstrating the suitability of LP-WGS for population genomics in resource-limited settings. Marked differences in genomic diversity were observed among ecotypes: midland chickens showed the highest nucleotide diversity (π = 0.00267), followed by lowland (π = 0.00233), whereas highland chickens showed the lowest diversity (π = 0.00203) and elevated genomic inbreeding (FROH and FHOM ≈ 0.18). Population structure analyses revealed clear genetic separation among ecotypes. PCA (13.91% variation explained) distinguished lowland chickens along PC1 and separated highland from midland along PC2, while ADMIXTURE and FST patterns supported three major ancestral genomic backgrounds. Functional annotation of private missense variants uncovered distinct adaptive signatures reflecting the contrasting agroecological conditions. Highland chickens showed enrichment of candidate genes potentially involved in physiological processes relevant to high-altitude environments, including cold response, angiogenesis, cardiovascular regulation and metabolic homeostasis (eg., PARP1, ACOX2, ITGB3, EDNRB, SOX8, and SOX10). Midland chickens exhibited candidate signals of selection in genes with known roles in innate antiviral immunity, bacterial defence and inflammatory regulation (eg., BAK1, CLSTN1, CYSLTR1, CYSLTR2, CXCR7, GIPR, DSCAM, GDAP1, TLR3, TLR4, TLR7, IFIH1, ADORA1, EPHB1, and TMPRSS2). Lowland chickens displayed candidate variants associated with heat-stress response, DNA damage repair, oxidative balance and cardiovascular support under extreme temperatures (e.g., MLH1, BDKRB1, GPR19, FLT1, CCL18, TGM2, and RAMP3). Overall, the results indicate substantial genomic differentiation among ecotypes and suggest candidate environment-associated genetic divergence across Tigray's diverse agroecological zones. These populations may represent important reservoirs of adaptive genetic variation for climate-resilient poultry breeding, warranting further functional validation and conservation-oriented management.

Animals

Whole-genome sequencing identifies genetic diversity and adaptive signatures of hypoxia and ultraviolet radiation in Chinese chickens.

INTRODUCTION: Domestic chickens primarily descended from the wild red junglefowl, play a crucial role in global egg and meat production. China hosts diverse indigenous chicken populations that have adapted to various environmental conditions, including high-altitude with hypoxic and ultraviolet radiation stress. METHOD: We analyzed whole-genome sequences of 118 birds from five Indigenous Chinese chicken populations and 295 chicken genomes from publicly available databases to identify genomic diversity, admixture, and selection signatures of chickens adapted to high-altitude environments. Selection signatures were identified using nucleotide diversity (π), Tajima's D, XPEHH, and XP-CLR, selection scan methods. RESULTS: We observed a reduction in genetic diversity and historical declines in effective population size in high-altitude chicken, suggesting ongoing selection pressures shaping these populations. Selection scans identified nine genomic regions under strong positive selection, enriched for genes associated with hypoxia and ultraviolet radiation. Notably, five genes (TPK1, BAZ2B, MARCHF7, LLGL2, and RCAN3) were repeatedly detected across multiple selection signature analyses. RNA-seq analysis further confirmed the differential expression of these genes in the lung and heart tissues of chickens adapted to high and low altitudes, reinforcing their role in physiological adaptation to hypoxic environments. Altitude adaptation is driven by the selection of genes involved in oxygen metabolism, cellular stress response, and energy regulation. CONCLUSION: Our study provides compelling genetic evidence for differentiation between high and low and high-altitude Chinese chicken populations. These findings also ensure our understanding of local adaptation in poultry and establish a genomic framework for breeding strategies to improve environmental resilience to altitude-related stressors.

Animals

Age-dependent resistance of chicken of Salmonella in vitro: antibacterial activity of lysed granule fraction of splenic adherent cells.

Lysed granule fraction was prepared from splenic adherent cells from 0-day-old to 2-month-old chickens and was examined antibacterial activity to Salmonella pullorum and Salmonella sentfenberg. The activity of lysed granule fraction from 2-month-old chickens was exhibited at pH 5.0 but not exhibited at pH 6.0 or 7.0 to both serotypes of Salmonella. At pH 5.0, lysed granule fraction from 0-day-old chickens revealed little antibacterial activity to S. pullorum but the activity increased with increasing age. At 240 min of incubation, 60% and 95% of S. pullorum were inactivated in lysed granule fractions from 7-day-old and 2-month-old chickens, respectively. High antibacterial activity to S. senftenberg was observed in lysed granule fraction from 2-month-old chickens in which 99.6% of the bacteria was inactivated during 240 min; In lysed granule fractions from 0-day old and 7-day-old chickens, 90% of S. senftenberg was inactivated during 60 min but the number of bacteria increased conversely after incubation for 240 min. Changes in enzymatic activities of 4 enzymes in the lysed granule fractions from various ages of chickens were examined. beta-Glucronidase increased statistically significantly to 2.4 and 7.6 times from 0-day-old to 7-day-old and 2-month-old chickens, respectively. Acid phosphatase and alkaline phosphatase increased slightly but not significantly different, and enzymatic activity of myeloperoxidase was scarcely detected.

Age Factors

Oxidative metabolism of chicken polymorphonuclear leucocytes during phagocytosis.

The oxidative response to phagocytosis by chicken polymorphonuclear leucocytes was investigated as compared to guinea pig polymorphonuclear leucocytes. The polymorphs from both species respond to phagocytosis with an increased oxygen consumption, an increased generation of O2 and H2O2, and an increased oxidation of glucose through the hexose monophosphate shunt. The rate of oxygen consumption, and generation of O2- and H2O2 by phagocytosing chicken polymorphonuclear leucocytes is considerably lower than with phagocytosing guinea pig polymorphonuclear leucocytes. By contrast, the extent of hexose monophosphate shunt stimulation in chicken polymorphs is comparable to that of guinea pig polymorphs. Evidence is presented suggesting that H2O2 is preferentially degraded in chicken cells through the glutathione cycle, whereas catalase and myeloperoxidase are the two main H2O2 degrading enzymes in guinea pig cells. The 20,000 g fraction of the postnuclear supernatant of chicken polymorphs contains a cyanide-insensitive NADPH oxidizing activity which is stimulated during phagocytosis. Similar properties for the NADPH oxidizing activity of guinea pig polymorphs have been previously reported. It is concluded that the metabolic burst of phagocytosing chicken polymorphonuclear leucocytes is qualitatively similar to that of guinea pig polymorphonuclear leucocytes, but the latter cells are more active in all the biochemical parameters that have been measured. The difference in the H2O2 degradation pathways between the two species is accounted for by the lack of myeloperoxidase and catalase in chicken polymorphs.

Animals

Breed classification of Lao People's Democratic Republic (Lao PDR) and Thai native chickens using synchrotron radiation-based Fourier transform infrared spectroscopy and genotyping by sequencing.

Lao PDR harbors substantial genetic diversity in native chicken populations, representing an important resource for sustainable production and long-term food security. This study aimed to classify five Lao native chicken breeds-Ou, Black Bone, Horn Chou, Yolk, and Chae-and to discriminate them from a Thai native breed, Leung Hang Khao (LK), using integrative genotype-based approaches. Blood samples were collected from 50 LK and Lao native chickens (32 Ou, 10 Black Bone, 9 Horn Chou, 121 Yolk, and 41 Chae). Genomic DNA was extracted and analyzed using synchrotron radiation-based Fourier-transform infrared (SR-FTIR) spectroscopy to characterize biochemical composition, while genotyping-by-sequencing (GBS) was employed to identify genome-wide single nucleotide polymorphisms (SNPs). SR-FTIR analysis revealed highly significant differences among breeds in nucleotide-associated functional groups, including thymine, adenine, guanine, cytosine, as well as DNA backbone and deoxyribose components (P < 0.001). Multivariate analyses demonstrated that principal component analysis (PCA) of SR-FTIR spectra effectively discriminated chicken breeds, while hierarchical cluster analysis (HCA) further resolved them into two major clusters with distinct sub-clusters, reflecting variation in DNA biochemical composition. In contrast, GBS analysis identified 1484 common SNPs; however, PCA based on SNP data showed limited resolution in clearly separating breeds, despite revealing similar clustering trends. Overall, the results highlight the strong discriminatory power of SR-FTIR spectroscopy for rapid and effective classification of native chicken breeds at the molecular level, outperforming SNP-based differentiation under the current marker density. This study provides novel insights into the application of synchrotron-based spectroscopic techniques in poultry genetics and contributes valuable baseline information for the conservation and utilization of Lao native chicken genetic resources.

Breed classification

Identification and functional characterization of a novel antiviral chicken interferon-&#x3c5;.

Interferons are critical mediators of antiviral immunity in vertebrates. While type IV interferon (IFN-&#x3c5;) has been identified in fish and amphibians, its existence and function in chickens remained unknown. Through systematic genomic screening, we identified and cloned a novel chicken interferon gene, designated ChIFN-&#x3c5;. Phylogenetic analysis placed ChIFN-&#x3c5; within a distinct clade alongside zebrafish and clawed frog IFN-&#x3c5;, confirming its identity as a type IV interferon, with minimal homology to classical type I, II, or III IFNs. Expression profiling revealed constitutive ChIFN-&#x3c5; expression in mucosal and immune tissues of healthy chickens, exhibiting a distinct developmental shift: highest in trachea and small intestine in 1-day-old chicks, shifting to spleen and lung in 4-week-old chickens. ChIFN-&#x3c5; expression was strongly upregulated following H9N2 AIV infection. Functionally, recombinant ChIFN-&#x3c5; protein activated the interferon-stimulated response element (ISRE) and Mx promoter in a dose-dependent manner and significantly inhibited the replication of both vesicular stomatitis virus (VSV) and H9N2 AIV in DF-1 cells. In vivo, early treatment with exogenous ChIFN-&#x3c5; significantly reduced pulmonary and tracheal viral loads and decreased oropharyngeal and cloacal virus shedding in H9N2-infected chickens. In conclusion, this study identifies and functionally characterizes the type IV interferon in chickens, elucidating the evolutionary status, regulated expression, and antiviral efficacy of ChIFN-&#x3c5;. These findings highlight its potential as a candidate for developing interferon-based therapies against avian viral diseases.

Animals

Isolation and identification of a highly oncogenic subgroup J avian leukosis virus strain from Chinese black chickens.

Avian leukosis virus subgroup J (ALV-J), an oncogenic retrovirus, is a highly contagious pathogen that induces myelocytomas, hemangiomas, and other neoplastic diseases in chickens. Recently, ALV-J infection in Chinese local chicken breeds has increased, with enhanced pathogenicity, posing a severe threat to the local poultry industry. In March 2024, a tumor outbreak occurred on a Chinese black chicken farm in Heze City, China, causing lethargy, emaciation, and tumorigenesis in the eyes and legs of affected chickens. Necropsy revealed extensive yellow-white neoplasms disseminated across the sternum, ribs, vertebrae, and visceral organs. Pathological examinations revealed multiple tumor types, including myelocytomas, hemangiomas, fibromas, and reticulosarcomas in the affected chickens. In this study, nine ALV-J isolates were isolated and designated HZ0319-1 to HZ0319-9. All isolates possessed an identical full-genome length of 7,687 bp and exhibited high nucleotide sequence similarity, ranging from 99.8% to 99.9%, suggesting that they were closely related variants from the same outbreak. Nucleotide sequence analysis identified unique deletions and mutations in the gag, pol, and gp85 genes, leading to predicted conformational changes in the P2, P10, and SU proteins. In addition, HZ0319 showed deletions in the r-TM region and a large 125-nt deletion in the E element of the 3' untranslated region, leaving only a short conserved fragment. The representative isolate HZ0319-1 showed stronger replication capacity than the reference strain NX0101 in DF-1 cells. In experimentally infected chicks, HZ0319-1 induced myelocytomas and hemangiomas and produced high viral load in multiple tissues. Notably, viral load analysis revealed that the liver showed the highest viral load at 1 day of age, whereas most other tissues reached peak viral load at 21 days of age, suggesting early hepatic replication followed by systemic dissemination. Furthermore, HZ0319-1infection significantly upregulated tumor-related host genes, including p53, c-Myc, c-Fos, and ZIC1, in tumor-associated tissues. These results demonstrate that the HZ0319-1 isolate has enhanced tumorigenicity and replication ability. Nucleotide mutations and deletions in both coding and non-coding regions of the viral genome may alter the viral tissue tropism and oncogenic potential. This study provides novel insights into the molecular characteristics and pathogenicity of ALV-J in local Chinese chickens, and provides a foundation for the prevention and eradication of ALV-J in the local poultry industry.

Avian leukosis virus subgroup J

Research note: Efficient preservation of genome-edited chicken germplasm via N-methylacetamide-based semen cryopreservation.

The rapid development of genome-edited chicken lines for agricultural and biomedical use requires effective methods for long-term preservation. In birds, cryopreservation of oocytes and embryos is challenging due to the structure of the egg. This makes semen cryopreservation one of the most practical alternatives, however, this method has not been validated in genome-edited chickens. This study evaluated the feasibility of cryopreserving semen from genome-edited chickens using an N-methylacetamide (N-MA)-based protocol. Two genome-edited chicken lines were used in this study which are a DAZL-GFP germ cell reporter line and a RAG1 knockout line. Semen was stored for either one week or one month prior to assessing fertility and hatchability. The results demonstrated that cryopreserved semen from both genome-edited chicken lines maintained their fertilizing capacity, with no significant differences in fertility or hatchability compared to cryopreserved wild-type groups at either storage duration. Hatchability remained above 80% across all groups, indicating that semen cryopreservation did not significantly affect embryo development after successful fertilization. Furthermore, SYBR-14/PI staining revealed no significant differences in sperm viability between genome-edited and control groups, although both showed a significant decrease in viability compared to fresh semen. These findings demonstrate that N-MA-based semen cryopreservation provides a reliable and practical method for the preservation of genome-edited chicken lines.

Chicken

The fate of antibiotics and antibiotic resistance genes in Large-Scale chicken farm Environments: Preliminary view of the performance of National veterinary Antimicrobial use reduction Action in Guangdong, China.

In 2018, China implemented the Veterinary Antimicrobial Use Reduction Action to curb the rapid development of antibiotic resistance (AR). However, the AR-related pollutions in animal farms after the reduction policy has been poorly investigated. Here, we performed a comprehensive investigation combining UPLC-MS/MS, metagenomic, and bacterial genomic analyses in eight representative large-scale chicken farms in Guangdong, China. Our results showed that antibiotics and ARGs contaminations were more severe in broiler farms than in layer farms. Notably, diverse tet(X) variants were prevalent in the chicken farms. These tet(X)s was carried by diverse E. coli lineages and obviously correlated with ISCR2 and IS1B transposases. The resistomes in chicken farms was significantly correlated with microbial community, and multiple factor analyses indicated that the joint effect of antibiotics-microbial community-MGEs was the most dominant driver of ARGs. Host tracking identified a variety of ARG bacterial hosts and the co-occurrence of ARGs-MRGs-MGEs. Source tracking indicated that the inherent component represented the main feature of resistomes in different hosts, while ARG transfer between the chicken gut and farm environments were frequent. A multiperspective evaluation of AR risk revealed that the early effect of antibiotic reduction was exhibited by the mitigation of maximum level of risky ARGs, prevalence of environmental AR pathogens, and HGT potential of ARGs mediated by phage structures. Overall, our findings provide insights into the antibiotic and ARG profiles in large-scale chicken farms with different rearing strategies and demonstrate a preliminary view of the performance of antibiotic reduction actions in China.

Chickens

Whole mitogenome profile of Pelung and Sentul chickens to reveal potency of Indonesian livestock genetic resources.

Native chickens are essential genetic resources in Indonesia, providing economic, cultural, and nutritional value with strong adaptability to local environments. Among these native chickens, Sentul and Pelung are recognized as national genetic resources due to dual-purpose and ornamental traits, respectively, but their whole mitogenome characterization remains limited. Therefore, this study aimed to assemble and analyze the whole mitogenome of Sentul and Pelung chickens as well as perform a comparison with 35 additional genomes from domestic chickens and jungle fowls across Asia. The experiment was carried out using next-generation sequencing and bioinformatics-based genome assembly and analysis. The results showed that both Sentul and Pelung genomes were 16,784&#xa0;bp in length and contained the typical mitochondrial gene composition, including 13 protein-coding genes (PCGs), 22 transfer ribonucleic acids (tRNAs), two ribosomal ribonucleic acids (rRNAs), and a control region (D-loop). Furthermore, comparative analysis identified five single-nucleotide polymorphisms (SNPs) distinguishing the two breeds, located in ND1, COX1, COX2, ND4, and the D-loop region. Phylogenetic reconstruction based on whole mitochondrial sequences showed that Sentul and Pelung chickens belong to haplogroup D, alongside other native breeds and red jungle fowls from Indonesia and the Philippines.

Genetic diversity

Resilience indicator traits in chickens: a systematic review.

The resilience of an animal is its ability to cope with short-term disturbances, including those caused by pathogens, through response and rapid recovery to its original state. Here, a systematic literature review was conducted to identify resilience indicators that have already been studied and implemented for chickens, along with the contexts or specific stressor under which they were examined. The literature review was based on predefined search criteria, including 'chickens' as the population of interest, a stress exposure description and the term 'resilience'. Screening of titles and abstracts was assisted by the AI-based tool ASReview, followed by full text screening and analysis. According to selection criteria, we finally identified 33 relevant publications on resilience indicator traits used in chickens. Two of these studies analyzed chicken resilience based on routinely collected big data, while the remaining majority were small or medium scale studies and trials. The majority of the studies focused on immune (n = 13) or thermal challenges (either before or after hatching; n = 14), especially heat stress. A variety of resilience indicators were studied; most studies investigated production- or performance-related parameters and/or immunity or disease-related parameters. About half of the studies included genetics- or gene expression-related indicators. Investigation of behavioral indicators of resilience - other than feed intake - was limited (n = 4). Overall, this systematic review provides a comprehensive overview of published resilience indicators in chickens and highlights gaps for future research. This review also revealed a need for the controlled use of terms like resilience, robustness, resistance, tolerance or adaptability, and the relevance of assessing the phase of recovery after a short-term disturbance in the context of resilience.

Animals

Comprehensive profiling of antibiotic resistance genes and functional clusters of orthologous groups annotation of gut microbiota in Indonesian Kedu chickens.

Antibiotic resistance is a growing global health concern, with poultry systems acting as important reservoirs of antibiotic resistance genes (ARGs). However, resistome and functional profiles of indigenous chickens raised under traditional systems remain underexplored. This study aimed to characterize the antibiotic resistome, virulence factor genes, and metabolic potential of gut microbiota in Indonesian Kedu chickens using a shotgun metagenomic approach. Digesta samples from five gastrointestinal segments of 21 healthy adult chickens were analyzed through high-throughput sequencing. ARGs were identified using the Comprehensive Antibiotic Resistance Database (CARD) and Antibiotic Resistance Genes Databases (ARDB), while virulence factors and functional genes were annotated using Virulence Factor Database (VFDB), Clusters of Orthologous Groups (COG), and Carbohydrate-Active EnZymes (CAZy) databases. Results revealed a diverse resistome dominated by multidrug resistance and efflux pump mechanisms, with prominent genes associated with fluoroquinolone, tetracycline, &#x3b2;-lactam, and glycopeptide resistance. The detection of clinically relevant ARGs suggests that genetic determinants associated with antimicrobial resistance are present in the gut microbiota of traditionally raised Kedu chickens, although metagenomic data alone cannot determine whether these genes are actively expressed or confer phenotypic resistance. Virulence factor analysis showed functions related to adherence, immune evasion, iron acquisition, quorum sensing, and efflux activity, reflecting strong microbial adaptability. Functional profiling demonstrated enrichment in translation, carbohydrate and amino acid metabolism, genome maintenance, and cell envelope biogenesis. Additionally, CAZyme analysis indicated a high capacity for complex polysaccharide degradation, supporting efficient utilization of fiber-rich traditional diets. In conclusion, this study provides a comprehensive metagenomic overview of antibiotic resistance and functional potential in Kedu chicken gut microbiota, emphasizing the importance of incorporating indigenous poultry into antimicrobial resistance surveillance within a One Health framework.

Antibiotic resistance genes

Deletion of Indian hedgehog gene causes dominant semi-lethal Creeper trait in chicken.

The Creeper trait, a classical monogenic phenotype of chicken, is controlled by a dominant semi-lethal gene. This trait has been widely cited in the genetics and molecular biology textbooks for illustrating autosomal dominant semi-lethal inheritance over decades. However, the genetic basis of the Creeper trait remains unknown. Here we have utilized ultra-deep sequencing and extensive analysis for targeting causative mutation controlling the Creeper trait. Our results indicated that the deletion of Indian hedgehog (IHH) gene was only found in the whole-genome sequencing data of lethal embryos and Creeper chickens. Large scale segregation analysis demonstrated that the deletion of IHH was fully linked with early embryonic death and the Creeper trait. Expression analysis showed a much lower expression of IHH in Creeper than wild-type chickens. We therefore suggest the deletion of IHH to be the causative mutation for the Creeper trait in chicken. Our findings unravel the genetic basis of the longstanding Creeper phenotype mystery in chicken as the same gene also underlies bone dysplasia in human and mouse, and thus highlight the significance of IHH in animal development and human haploinsufficiency disorders.

Animals

The presence of immunoregulatory cells in chicken thymus: function in B and T cell responses.

The presence of immunoregulatory cells in chicken thymus was studied by using several different systems. Chickens injected with large numbers of syngeneic thymocytes were tested for their ability to produce antibody to heterologous red cells. Similar chickens were studied for their ability to reject allogeneic skin grafts. In separate studies, mixtures of thymocytes with spleen cells or with peripheral blood leukocytes were assayed for their ability to respond to PHA or to produce a graft-vs-host reaction in embryonic chicks. These studies indicated that immunoregulatory cells exist in chicken thymus, which displays both helper and suppressor activity. The suppressor cells were more prevalent or more easily detectable in young birds and in chickens with intact bursas. The helper function of thymocytes was seen to better advantage with cells derived from older animals and from bursectomized donors.

Aging

Renal transport and renal metabolism of 2, 4, 5-trichlorophenoxyacetate by the chicken.

In vitro and in vivo renal tubular transport of 2,4,5-trichlorophenoxyacetate (2,4,5-T), a hormone-type herbicide, was studied in the chicken. Renal cortical slices incubated with 14C-labeled 2,4,5-T demonstrated a slice-to-medium ratio of the 14C-label of 26 after 2 hr of incubation. This accumulation was inhibited significantly by probenecid, phenol red, dinitrophenol and iodoacetamide. In vitro metabolism of 2,4,5-T was apparent in the renal slice studies; at least two metabolites were found by electrophoretic analysis. Even though chicken liver slices were able to accumulate 2,4,5-T, the accumulated herbicide was not metabolized. In addition, chicken kidney slices metabolized 2,4,5-T to a much greater extent than did rat or rabbit kidney slices, which produced only negligible amounts of metabolites. When [14C]-2,4,5-T was infused into the renal portal system of the chicken, the 14C-label originally associated with 2,4,5-T was shown to be secreted by the renal tubule. Probenecid inhibited the transport of the 14C-label without affecting the simultaneous transport of tetra-ethylammonium, an organic cation transported by the renal proximal tubule. At the present infusion rate of 2,4,5-T (2.9 X 10(-10) mol/min), the excreted label was associated primarily with an acidic metabolite(s). It can be concluded that the chicken transports 2,4,5-T by a probenecid sensitive mechanism; however, this transport appears to be associated with the metabolism of the herbicide.

2,4,5-Trichlorophenoxyacetic Acid