Poultry Science最新文献

Achieving stable and efficient transgene expression is a key challenge in advancing avian genome engineering. Although viral vector-based and piggyBac-mediated transgenesis have been widely used in chickens, both approaches are prone to epigenetic silencing, leading to inconsistent, tissue-specific, and often diminished expression over time. This variability limits used of transgenes requiring robust and long-term expression across multiple tissues. In mammals, site-specific integration into genomic safe harbor loci, such as Rosa26, has enabled stable and predictable transgene expression without disrupting endogenous gene function; however, such strategy has not been established in birds. In this research, we hypothesized that integrating Cas9 into endogenous housekeeping genes (the ACTB and GAPDH) could achieve efficient gene editing in chickens through stable and ubiquitous transgene expression. Using two different approaches, 3'-targeted gene insertion and gene tagging, we inserted Cas9 and GFP cassettes into defined genomic loci in chicken DF-1 cells. Both approaches exhibited stable expression of transgenes in the cells, and functional assays confirmed that Cas9 showed highly efficient nuclease activity following guide RNA delivery. Additionally, we derived single-cell clones stably expressing Cas9, enabling uniform and reproducible genome editing in downstream applications. Targeted insertion of transgenes into active housekeeping genes as candidate safe harbor loci mitigates the limitations of random integration and promoter silencing, offering a robust platform for consistent transgene expression in poultry biotechnology and genome engineering.

Avian fatty liver disease is a metabolic disease characterized by hepatocellular steatosis caused by fat deposition due to lipid metabolism disorders in poultry. AdipoRon, an adiponectin receptor agonist, has various biological effects, such as regulating glucose and lipid metabolism disorders, increasing insulin sensitivity, improving liver fat accumulation, and preventing inflammation and oxidative stress. Our previous study revealed that AdipoRon protected against liver injury induced by a high-fat diet and lipopolysaccharide in poultry. In this study, leghorn male hepatoma (LMH) cells were used to construct a lipotoxic injury model with mixed fatty acids (oleic acid + palmitic acid), and AdipoRon was subsequently used to intervene, followed by inhibition and activation of AMPK signaling pathways using an antagonist and agonist of AMPK, respectively, to detect lipid content and lipid deposition, hepatocyte injury-related transaminase activity, and the expression levels of lipid metabolism-related genes and key signaling molecules that regulate lipid metabolism, as well as the cellular lipid composition in LMH cells. AdipoRon promoted fatty acid oxidation, reduced lipid synthesis and deposition, and alleviated mixed fatty acid-induced lipotoxic injury through the regulation of the expression of adiponectin receptors, AMPK, PPARα, and key genes involved in lipid metabolism. The inhibition or activation of AMPK signaling pathways could regulate the expression of AdipoR1, AdipoR2, AMPK and p-AMPK, thereby altering the expression of lipid metabolism-related genes and antagonizing or synergistically increasing the ameliorative effects of AdipoRon on cellular lipid metabolism disorders, lipid deposition and cell injury. Lipidomic analysis further suggested that AdipoRon could regulate the metabolism of lipids such as sphingolipids, glycerophospholipids, acylcarnitines, and glycerolipids; reduce the accumulation of lipids such as ceramides, sphingomyelins, triacylglycerol, and acylcarnitines; maintain the metabolic homeostasis of phosphatidylamine and phosphatidylcholine, as well as cell membrane structural integrity and functional stability; and mitigate lipotoxic injury in LMH cells. This study provides new insights into targeted interventions involving adiponectin and its receptors to prevent and treat avian fatty liver disease.

The H1N1 influenza virus is a major pandemic and seasonal pathogen with a broad host range, posing a substantial threat to human health and underscoring the need for continuous surveillance. Wild birds, as natural reservoirs of avian influenza viruses (AIVs), carry H1N1 strains capable of reassorting with other influenza viruses, which can drive pandemic emergence. The global migration of wild birds facilitates the spread of these viruses, and their interactions with poultry increase the risk of cross-species transmission, further amplifying the public health threat. However, knowledge of H1N1 genetic diversity in wild birds remains limited. Database analysis shows 80% of avian-origin H1N1 isolates come from wild birds across over 40 countries, mainly in North America, Europe and Asia. This study characterized the molecular traits and genetic evolution of four H1N1 AIVs isolated from common teal and spot-billed ducks during 2019-2021. Phylogenetic and sequence analyses revealed these viruses cluster into distinct lineages, divergent from mammalian H1N1 strains, with complex genetic origins involving frequent recombination and high diversity. Frequent wild bird-poultry transmission elevates zoonotic risks. Our findings highlight wild birds' critical role in H1N1 transmission and confirm their role as an H1N1 gene pool, emphasizing the need for sustained monitoring and research.

Eukaryotic initiation factors (eIFs), a bunch of proteins that deeply involved in translation initiation of mRNA in eukaryotes, are closely associated with physiological and pathological processes. eIF3m, a core subunit in eIF3 complex, also played critical roles in virus infection. In this study, a co-immunoprecipitation coupled with mass spectrometry (Co-IP/MS) was performed to identify host factors that interacted with ORF1B, a unique non-structural protein of fowl adenovirus serotype 4 (FAdV-4). Among 2502 cellular proteins, eIF3m, especially its C-terminal part, was verified to interact with ORF1B and these two proteins co-localized in the cytoplasm. Importantly, overexpression of eIF3m promoted FAdV-4 replication in LMH cells, whereas knockdown eIF3m exerted an opposite effect. Collectively, these findings indicate that ORF1B hijacked eIF3m to positively participate in FAdV-4 infection.

Salmonella enterica is a major foodborne pathogen associated with poultry, representing a critical challenge for food safety worldwide. Accurate identification of serovar diversity is essential for designing control strategies; however, conventional culture-based methods often underestimate this complexity. In this study, we report the first application of CRISPR-SeroSeq in Ecuador to characterize Salmonella serovar diversity in commercial broilers. A total of 76 flocks (one hose of one farm in different cycles) originated across 19 broiler farms were sampled. All flocks belonged to an integrated poultry company. From all samples, 77.6% tested positive for Salmonella. CRISPR-SeroSeq analysis revealed a clear dominance of serovar Infantis, even within mixed populations. Importantly, serovars of significant public health concern, including Enteritidis and Typhimurium, were detected at low frequencies that would likely be missed by conventional methods. These findings highlight the utility of high-resolution serotyping approaches, providing valuable insights for targeted interventions to improve poultry production biosecurity and food safety.

Like mammals, the avian intestinal epithelium is innervated by three neuronal pathways: vagal and sympathetic terminals, which originate from ganglia outside the gut wall and send information to the brain to modulate visceral sensitivity, appetite, and gut homeostasis; and the enteric nervous system (ENS), a complex network embedded within the gut wall that functions independently from the brain. The ENS coordinates essential GI physiological functions, such as intestinal motility, peristalsis, digestion, and absorption of nutrients and water. Recent studies conducted in mammals have shown that enteric neurons can orchestrate the intestinal immune response and reduce Salmonella colonization in the GI tract. However, such neuronal-mediates defense mechanisms have not yet been explored in the poultry gut. This review will provide a comprehensive overview of the avian ENS, highlighting similarities and differences with the well-known mammalian ENS. Additionally, particular focus will be given on ENS-dependent neuroimmune interactions that could reveal novel biomolecular mechanisms to mediate health, disease susceptibility, behavior, and other aspects as affected by the chicken GI tract.

Bone damage is an important welfare issue in the poultry industry, yet large-scale phenotyping of chicken bone strength currently relies on time-consuming manual annotation of X-rays or destructive post-mortem testing. To address this, an end-to-end deep-learning pipeline was developed that automatically (i) segments the chicken tibiotarsus from lateral X-ray images (U-Net, Dice = 0.91) and (ii) predicts its breaking strength from pixel intensities alone. Using 916 curated bone images, the predictor achieved moderately high correlation with measured breaking strength (maximum Pearson's correlation of 0.74), exceeding the performance of a previous labour-intensive manual annotation method. Image-derived predictions were moderately heritable (h² ≈ 0.16) and exhibited an exceptionally high genetic correlation with the physical trait, indicating that selection on the model-derived phenotype is a good proxy to select for bone strength. The workflow therefore provides a potential rapid, non-invasive and genetically informative alternative to post-mortem testing, paving the way for the routine incorporation of bone-quality traits into commercial breeding programmes and improved poultry welfare at scale.

Mycoplasma gallisepticum (MG) is a primary avian pathogen that causes chronic respiratory disease, leading to significant economic losses in the poultry industry. This study aimed to investigate the pharmacokinetic/pharmacodynamic (PK/PD) relationship of a novel pleuromutilin derivative, 14-O-[(4-amino-6-hydroxy-pyrimidine-2-yl) thioacetyl] mutilin (APTM), against MG in a chicken infection model to provide a basis for a rational dosage regimen. The in vitro activity of APTM against MG strain S6 was assessed by determining the minimum inhibitory concentration (MIC) and time-kill kinetics. An intratracheal MG infection model was established in chickens. The pharmacokinetic profile was evaluated after single oral administrations of APTM at 5, 15, and 40 mg/kg. The pharmacodynamic efficacy was determined by quantifying the bacterial reduction in the lungs after three consecutive days of oral treatment with doses ranging from 0 to 40 mg/kg. The PK/PD data were integrated and analyzed using an inhibitory sigmoid Emax model. The MIC of APTM against MG S6 was 0.03125 µg/mL, and in vitro time-kill assays demonstrated concentration-dependent bactericidal activity. In chickens, APTM was rapidly absorbed (T_max: 0.25-0.5 h), with both C_max and AUC_0-24h exhibiting excellent dose proportionality (R² > 0.99) over the tested range. In the efficacy study, APTM produced a dose-dependent reduction in lung bacterial load, with a maximum mean reduction of 2.80 log₁₀CFU/mL observed at the 40 mg/kg dose, indicating a bactericidal effect. The PK/PD indices, AUC_0-24h/MIC and C_max/MIC, were both highly correlated with the in vivo antimicrobial effect (R² = 0.9424 and 0.9428, respectively). To achieve a 2-log₁₀CFU/mL reduction in bacterial load, the target AUC_0-24h/MIC value was determined to be 492.75, which corresponds to a calculated daily oral dose of 22 mg/kg. These findings demonstrate the potent efficacy of APTM against MG and provide a quantitative scientific foundation for its therapeutic use in poultry. Specifically, a daily oral dose of 22 mg/kg was identified as the breakpoint for a bactericidal effect (2-log10 reduction), suggesting APTM is a potent candidate for controlling MG infections in poultry.

Cadmium (Cd) induces oxidative stress and inflammation, leading to hepatotoxicity in animals. Honokiol (HNK) has gained much attention owing to its anti-inflammatory and antioxidant properties, and may offer protection against liver diseases. However, whether HNK can improve Cd-induced ultrastructural and functional variation in hepatocytes is largely unknown. In this study, day-old broiler were divided into four treatment groups including control/untreated group Cd (50mg/L), HNK (50mg/kg), and Cd+HNK (50mg/L+50mg/kg) for 42 days, respectively. In Silico analysis was conducted to reveal the potential interaction of HNK with Bax and Bcl-2 proteins to determine how HNK affected these two protein targets to mediate the effects of Cd toxicity. Results revealed that Cd exposure caused significant damage to the ultrastructure and functional activity of hepatocytes compared to the control group. Notably, HNK treatment helped recover and maintain the integrity of the nucleus and mitochondrial cristae in hepatocytes. In addition, HNK reduced oxidative stress with the increased activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and decreased malondialdehyde (MDA) content in the liver tissue. The HNK prevented lipid accumulation in the liver tissue induced by cadmium toxicity. Furthermore, HNK decreased the expression of apoptotic protein and gene expression of Caspase-3, and increased expression of the anti-apoptotic protein Bcl-2 by immunohistochemistry and qPCR. In conclusion, findings of the present study revealed the potential of HNK to alleviate Cd-induced ultrastructural and functional perturbations that cause hepatotoxicity in chicken, making it a promising therapeutic agent for Cd poisoning in animals.