Pub Date : 2026-01-28DOI: 10.1016/j.psj.2026.106539
Yongqing Cao, Tao Zeng, Li Chen, Jindong Ren, Yong Tian, Tiantian Gu, Wei Han, Jing Feng, Lili Xian, Shuangbao Gun, Lizhi Lu
The Tibetan chicken (Gallus gallus domesticus), a native breed inhabiting the Qinghai-Tibet Plateau, has developed remarkable tolerance to chronic hypoxia. However, the molecular and epigenetic mechanisms underlying its high-altitude adaptation remain unclear. In this study, we integrated genome, transcriptome, and DNA methylome data from Tibetan chickens (TC) and three low-altitude breeds. Principal component analysis revealed clear genetic, epigenetic, and transcriptional divergence between TC and lowland chickens. Cardiac enzyme assays showed significantly higher activities of LDH, SDH, SOD, CAT, and GSH-Px in TC (p < 0.05), indicating enhanced oxidative metabolism and antioxidant defense under hypoxia. Transcriptomic analysis identified 2,532 common differentially expressed genes (co-DEGs), with upregulated genes enriched in oxidative phosphorylation, fatty acid metabolism, and hypoxia response pathways. Integration with methylome data demonstrated a significant negative correlation between promoter methylation and gene expression. Among 144 genes showing promoter hypomethylation coupled with transcriptional activation, five key genes-PDK4, BNIP3L, ATG3, SLC7A5, and OMA1-were identified as central regulators of hypoxia adaptation, participating in metabolic reprogramming, mitochondrial homeostasis, and autophagy. Our findings reveal that promoter hypomethylation acts as a major epigenetic mechanism mediating transcriptional activation of hypoxia-responsive genes in Tibetan chickens. The coordinated regulation of energy metabolism, antioxidant defense, and mitochondrial quality control contributes to their physiological resilience in high-altitude environments. This study provides novel insights into the molecular and epigenetic basis of high-altitude adaptation in avian species and offers valuable references for hypoxia-resistance breeding in poultry.
{"title":"DNA methylation-mediated regulation of hypoxia-responsive genes facilitates high-altitude adaptation in Tibetan chickens.","authors":"Yongqing Cao, Tao Zeng, Li Chen, Jindong Ren, Yong Tian, Tiantian Gu, Wei Han, Jing Feng, Lili Xian, Shuangbao Gun, Lizhi Lu","doi":"10.1016/j.psj.2026.106539","DOIUrl":"https://doi.org/10.1016/j.psj.2026.106539","url":null,"abstract":"<p><p>The Tibetan chicken (Gallus gallus domesticus), a native breed inhabiting the Qinghai-Tibet Plateau, has developed remarkable tolerance to chronic hypoxia. However, the molecular and epigenetic mechanisms underlying its high-altitude adaptation remain unclear. In this study, we integrated genome, transcriptome, and DNA methylome data from Tibetan chickens (TC) and three low-altitude breeds. Principal component analysis revealed clear genetic, epigenetic, and transcriptional divergence between TC and lowland chickens. Cardiac enzyme assays showed significantly higher activities of LDH, SDH, SOD, CAT, and GSH-Px in TC (p < 0.05), indicating enhanced oxidative metabolism and antioxidant defense under hypoxia. Transcriptomic analysis identified 2,532 common differentially expressed genes (co-DEGs), with upregulated genes enriched in oxidative phosphorylation, fatty acid metabolism, and hypoxia response pathways. Integration with methylome data demonstrated a significant negative correlation between promoter methylation and gene expression. Among 144 genes showing promoter hypomethylation coupled with transcriptional activation, five key genes-PDK4, BNIP3L, ATG3, SLC7A5, and OMA1-were identified as central regulators of hypoxia adaptation, participating in metabolic reprogramming, mitochondrial homeostasis, and autophagy. Our findings reveal that promoter hypomethylation acts as a major epigenetic mechanism mediating transcriptional activation of hypoxia-responsive genes in Tibetan chickens. The coordinated regulation of energy metabolism, antioxidant defense, and mitochondrial quality control contributes to their physiological resilience in high-altitude environments. This study provides novel insights into the molecular and epigenetic basis of high-altitude adaptation in avian species and offers valuable references for hypoxia-resistance breeding in poultry.</p>","PeriodicalId":20459,"journal":{"name":"Poultry Science","volume":"105 4","pages":"106539"},"PeriodicalIF":4.2,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146126440","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-28DOI: 10.1016/j.psj.2026.106537
Sara E Cloft, Prafulla Regmi, Cara I Robison, Deana Jones, Darrin M Karcher
The transition from conventional cages to cage-free aviary systems in egg production presents unique challenges for performance, welfare, and skeletal health of laying hens. While extensive data exists for conventional systems, aviary systems require comprehensive investigation due to larger colony sizes and increased opportunities for vertical and lateral movement. This study evaluated the production, welfare, and skeletal characteristics of four commercial laying hen hybrids, two brown egg (brown) and two white egg (white) strains, in an aviary housing system under common management. Brown strains were consistently heavier with larger tibia volume, surface area, and mineral content compared to white strains. All 4 strains achieved at least 91 % hen day egg production, with white strain C having 8 percentage point higher production rates throughout the majority of lay. Feather coverage deteriorated as all hens aged, but white strains, especially strain C, had more frequent feather damage during assessments. Brown strains had more incidence of keel damage based on manual palpation. However, visual inspection of excised keel bones revealed brown strain B had fewer fractures than all other strains, though 90 % of the keel bones had fractures, frequently in the tip. These findings reveal significant strain-specific differences in production, skeletal health, and welfare in aviary systems. Our use of common management may have hindered hens from achieving their full genetic potential; thus, tailoring management and housing practices to accommodate these differences is crucial for successful cage-free egg production and hen welfare.
{"title":"Performance, Skeletal traits, and welfare indicators of four laying hen strains in aviary housing under common management.","authors":"Sara E Cloft, Prafulla Regmi, Cara I Robison, Deana Jones, Darrin M Karcher","doi":"10.1016/j.psj.2026.106537","DOIUrl":"https://doi.org/10.1016/j.psj.2026.106537","url":null,"abstract":"<p><p>The transition from conventional cages to cage-free aviary systems in egg production presents unique challenges for performance, welfare, and skeletal health of laying hens. While extensive data exists for conventional systems, aviary systems require comprehensive investigation due to larger colony sizes and increased opportunities for vertical and lateral movement. This study evaluated the production, welfare, and skeletal characteristics of four commercial laying hen hybrids, two brown egg (brown) and two white egg (white) strains, in an aviary housing system under common management. Brown strains were consistently heavier with larger tibia volume, surface area, and mineral content compared to white strains. All 4 strains achieved at least 91 % hen day egg production, with white strain C having 8 percentage point higher production rates throughout the majority of lay. Feather coverage deteriorated as all hens aged, but white strains, especially strain C, had more frequent feather damage during assessments. Brown strains had more incidence of keel damage based on manual palpation. However, visual inspection of excised keel bones revealed brown strain B had fewer fractures than all other strains, though 90 % of the keel bones had fractures, frequently in the tip. These findings reveal significant strain-specific differences in production, skeletal health, and welfare in aviary systems. Our use of common management may have hindered hens from achieving their full genetic potential; thus, tailoring management and housing practices to accommodate these differences is crucial for successful cage-free egg production and hen welfare.</p>","PeriodicalId":20459,"journal":{"name":"Poultry Science","volume":"105 4","pages":"106537"},"PeriodicalIF":4.2,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146137681","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-28DOI: 10.1016/j.psj.2026.106534
Lingqi Li, Yang Ye, Zhen Li, Peng Wang, Xinglian Xu, Xiao Sun, Yulong Zhang, Qingqin Lv, Yujia Yang, Tianran Huang, Lei Wang, Zhixin Chen, Rongxin Yuan
Currently, palpation remains the predominant method for classifying wooden breast (WB). This approach requires considerable labor and time resources, and it fails to precisely characterize the complex internal structural distribution of the disease and lacks a rational utilization plan for WB-affected breast fillets. Thus, the scientific stratification and classification of WB must be investigated. This study aims to characterize WB severity using ultrasound-derived internal spatial information, combined with ImageJ threshold binarization and scale calibration to quantify the spatial extent of pathological features. Herein, chicken breast fillets from Arbor Acres broilers were collected (n = 240, males, 42 days old) and categorized into four categories: normal (NORM, n = 60), mild (MILD, n = 60), moderate (MOD, n = 60), and severe (SEV, n = 60) conditions. WB samples were classified via ultrasound scanning and deep learning (DL). MobileNetV3, ResNet18, and AlexNet achieved training accuracies of 99.50%, 96.62%, and 95.64%, respectively, with validation accuracies of 98.71%, 90.09%, and 92.95%. For the four aforementioned classifications, the MobileNetV3 model achieved accuracies of 95%, 100%, 100%, and 99%, respectively, and exhibited a precision of 98.25%, a recall of 98.22%, and an F1-score of 98.23%. Image analysis delineated boundaries between pathological regions and normal muscle tissues in WB, validated by bioimpedance and stress-strain measurements. Segmentation ranges for MILD, MOD, and SEV pathological severity were determined as approximately 55%, 62%, and 65%, respectively, marking the first precise internal stratification of WB. Results showed that ultrasound imaging combined with DL effectively assessed myopathy distribution within WB, enabling accurate classification and stratification for practical applications.
目前，触诊仍然是木质乳房（WB）分类的主要方法。这种方法需要大量的人力和时间资源，不能准确表征疾病复杂的内部结构分布，缺乏合理的利用计划。因此，必须对WB进行科学的分层和分类研究。本研究旨在利用超声来源的内部空间信息来表征WB严重程度，结合ImageJ阈值二值化和尺度校准来量化病理特征的空间范围。选取42日龄爱拔加（Arbor Acres）肉鸡鸡胸片240块，分为正常（NORM, n = 60）、轻度（mild, n = 60）、中度（MOD, n = 60）和重度（SEV, n = 60） 4类。WB样本通过超声扫描和深度学习（DL）进行分类。MobileNetV3、ResNet18和AlexNet的训练准确率分别为99.50%、96.62%和95.64%，验证准确率分别为98.71%、90.09%和92.95%。对于上述四种分类，MobileNetV3模型的准确率分别为95%、100%、100%和99%，准确率为98.25%，召回率为98.22%，f1得分为98.23%。图像分析描绘了病理区域和正常肌肉组织之间的边界，通过生物阻抗和应力-应变测量验证。MILD、MOD和SEV病理严重程度的分割范围分别约为55%、62%和65%，标志着WB首次精确的内部分层。结果显示，超声成像联合DL可有效评估肌病在WB内的分布，为实际应用提供准确的分类和分层。
{"title":"Deep learning-based classification and internal region stratification of wooden breast in broiler by using ultrasound imaging.","authors":"Lingqi Li, Yang Ye, Zhen Li, Peng Wang, Xinglian Xu, Xiao Sun, Yulong Zhang, Qingqin Lv, Yujia Yang, Tianran Huang, Lei Wang, Zhixin Chen, Rongxin Yuan","doi":"10.1016/j.psj.2026.106534","DOIUrl":"10.1016/j.psj.2026.106534","url":null,"abstract":"<p><p>Currently, palpation remains the predominant method for classifying wooden breast (WB). This approach requires considerable labor and time resources, and it fails to precisely characterize the complex internal structural distribution of the disease and lacks a rational utilization plan for WB-affected breast fillets. Thus, the scientific stratification and classification of WB must be investigated. This study aims to characterize WB severity using ultrasound-derived internal spatial information, combined with ImageJ threshold binarization and scale calibration to quantify the spatial extent of pathological features. Herein, chicken breast fillets from Arbor Acres broilers were collected (n = 240, males, 42 days old) and categorized into four categories: normal (NORM, n = 60), mild (MILD, n = 60), moderate (MOD, n = 60), and severe (SEV, n = 60) conditions. WB samples were classified via ultrasound scanning and deep learning (DL). MobileNetV3, ResNet18, and AlexNet achieved training accuracies of 99.50%, 96.62%, and 95.64%, respectively, with validation accuracies of 98.71%, 90.09%, and 92.95%. For the four aforementioned classifications, the MobileNetV3 model achieved accuracies of 95%, 100%, 100%, and 99%, respectively, and exhibited a precision of 98.25%, a recall of 98.22%, and an F1-score of 98.23%. Image analysis delineated boundaries between pathological regions and normal muscle tissues in WB, validated by bioimpedance and stress-strain measurements. Segmentation ranges for MILD, MOD, and SEV pathological severity were determined as approximately 55%, 62%, and 65%, respectively, marking the first precise internal stratification of WB. Results showed that ultrasound imaging combined with DL effectively assessed myopathy distribution within WB, enabling accurate classification and stratification for practical applications.</p>","PeriodicalId":20459,"journal":{"name":"Poultry Science","volume":"105 4","pages":"106534"},"PeriodicalIF":4.2,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12887160/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146113950","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Chicken livers can be sustainably developed into nutraceuticals through the circular-agriculture innovation chain. A supplement (GBHP01) formulated from chicken-liver hydrolysates contains free-type hypolipidemic amino acids (threonine, valine, leucine, isoleucine, and taurine) and the imidazole-ring dipeptide (anserine). In this study, mice were assigned to four groups: (1) Control: control diet (AIN-93M formula; fat providing 9.4% of total calories), (2) HFD: high-fat diet (fat providing 46.5% of total calories), (3) GBHP01.L: HFD supplemented with GBHP01 at 133.61 mg/Kg BW/day, and (4) GBHP01.H: HFD supplemented with GBHP01 at 267.22 mg/Kg BW/day. GBHP01 was administered by oral gavage. In 20-week HFD feeding, GBHP01 supplementation significantly reduced serum lipids, ALT, AST, and hepatic tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) concentrations, while enhancing reduced GSH, TEAC, catalase, and GSH-Px levels (p<0.05). Histological analysis revealed decreased hepatic lipid accumulation, associated with decreased diacylglycerol O-acyltransferase 2 (DGAT2) and increased acyl-CoA dehydrogenase medium chain (ACADM) expression. GBHP01 also promoted fecal bile acid and triglyceride excretion, indicating reduced fat absorption. Fecal microbiota profiling showed that HFD disrupted microbial diversity, increasing detrimental genera (Desulfovibrio, Bilophila, and Lachnoclostridium) while decreasing beneficial taxa (Lactobacillus and Akkermansia). GBHP01 may contribute to shifts in gut microbial composition by elevating probiotic species (L. reuteri and L. murinus) and reducing inflammatory taxa (Bilophila and Mucispirillum), suggesting its potential as a dietary intervention against HFD-induced metabolic disorders.
{"title":"Free amino-acid and imidazole-ring dipeptide profiles of chicken-liver-hydrolysate supplement and its modulatory effects on lipid metabolism, oxidative status, and inflammation in livers, as well as gut microbiota in a high-fat diet.","authors":"Yi-Ling Lin, Christoper Caesar Yudho Sutopo, Sheng-Yao Wang, Jr-Wei Chen, Yi-Feng Kao, Yi-Chen Chen","doi":"10.1016/j.psj.2026.106533","DOIUrl":"10.1016/j.psj.2026.106533","url":null,"abstract":"<p><p>Chicken livers can be sustainably developed into nutraceuticals through the circular-agriculture innovation chain. A supplement (GBHP01) formulated from chicken-liver hydrolysates contains free-type hypolipidemic amino acids (threonine, valine, leucine, isoleucine, and taurine) and the imidazole-ring dipeptide (anserine). In this study, mice were assigned to four groups: (1) Control: control diet (AIN-93M formula; fat providing 9.4% of total calories), (2) HFD: high-fat diet (fat providing 46.5% of total calories), (3) GBHP01.L: HFD supplemented with GBHP01 at 133.61 mg/Kg BW/day, and (4) GBHP01.H: HFD supplemented with GBHP01 at 267.22 mg/Kg BW/day. GBHP01 was administered by oral gavage. In 20-week HFD feeding, GBHP01 supplementation significantly reduced serum lipids, ALT, AST, and hepatic tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) concentrations, while enhancing reduced GSH, TEAC, catalase, and GSH-Px levels (p<0.05). Histological analysis revealed decreased hepatic lipid accumulation, associated with decreased diacylglycerol O-acyltransferase 2 (DGAT2) and increased acyl-CoA dehydrogenase medium chain (ACADM) expression. GBHP01 also promoted fecal bile acid and triglyceride excretion, indicating reduced fat absorption. Fecal microbiota profiling showed that HFD disrupted microbial diversity, increasing detrimental genera (Desulfovibrio, Bilophila, and Lachnoclostridium) while decreasing beneficial taxa (Lactobacillus and Akkermansia). GBHP01 may contribute to shifts in gut microbial composition by elevating probiotic species (L. reuteri and L. murinus) and reducing inflammatory taxa (Bilophila and Mucispirillum), suggesting its potential as a dietary intervention against HFD-induced metabolic disorders.</p>","PeriodicalId":20459,"journal":{"name":"Poultry Science","volume":"105 4","pages":"106533"},"PeriodicalIF":4.2,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12887177/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146119765","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The H7N9 subtype of avian influenza virus (AIV) poses a significant and ongoing threat to public health. As a critical structural and functional component, the viral nucleoprotein (NP) is abundantly expressed during the early stages of AIV replication; however, its interactions with host proteins and their functional consequences remain largely uncharacterized. This study aimed to identify the NP-host interaction and elucidate the mechanisms by which these interactions modulate AIV replication. Here, we employed mass spectrometry and identified the DEAD-box helicase 6 (DDX6) as a novel NP-interacting partner, an association found to be regulated by an interferon-stimulated gene (ISG15). The NP-DDX6 interaction was robustly validated by co-immunoprecipitation, immunofluorescence co-localization, bimolecular fluorescence complementation, and molecular docking assays. Functional investigations revealed that DDX6 acts as a potent negative regulator of AIV replication. Mechanistically, DDX6 not only impaired the nuclear import of NP and suppressed viral polymerase activity, but also stimulated the production of interferon (IFN)-α/β. This IFN-I induction, in turn, triggers the expression of downstream antiviral effectors such as ISG15. Furthermore, we uncovered that DDX6 fine-tunes this pathway by playing a sophisticated dual regulatory role: it enhances the pool of free, antiviral ISG15 monomers while concurrently reducing ISGylation via two deubiquitinases (USP16/USP18). Collectively, these findings not only establish DDX6 as a crucial host factor with potent antiviral activity but also enrich our understanding of host-virus interaction networks.
{"title":"The host RNA helicase DDX6 restricts avian influenza virus replication by targeting viral NP and modulating ISG15.","authors":"Xiaolong Lu, Chenyu Lu, Mengyang He, Xinen Tang, Zhuxing Ji, Hongqi Wu, Kaituo Liu, Wenhao Yang, Yu Chen, Ruyi Gao, Jiao Hu, Min Gu, Shunlin Hu, Xiaowen Liu, Xiaoquan Wang, Xiufan Liu","doi":"10.1016/j.psj.2026.106529","DOIUrl":"10.1016/j.psj.2026.106529","url":null,"abstract":"<p><p>The H7N9 subtype of avian influenza virus (AIV) poses a significant and ongoing threat to public health. As a critical structural and functional component, the viral nucleoprotein (NP) is abundantly expressed during the early stages of AIV replication; however, its interactions with host proteins and their functional consequences remain largely uncharacterized. This study aimed to identify the NP-host interaction and elucidate the mechanisms by which these interactions modulate AIV replication. Here, we employed mass spectrometry and identified the DEAD-box helicase 6 (DDX6) as a novel NP-interacting partner, an association found to be regulated by an interferon-stimulated gene (ISG15). The NP-DDX6 interaction was robustly validated by co-immunoprecipitation, immunofluorescence co-localization, bimolecular fluorescence complementation, and molecular docking assays. Functional investigations revealed that DDX6 acts as a potent negative regulator of AIV replication. Mechanistically, DDX6 not only impaired the nuclear import of NP and suppressed viral polymerase activity, but also stimulated the production of interferon (IFN)-α/β. This IFN-I induction, in turn, triggers the expression of downstream antiviral effectors such as ISG15. Furthermore, we uncovered that DDX6 fine-tunes this pathway by playing a sophisticated dual regulatory role: it enhances the pool of free, antiviral ISG15 monomers while concurrently reducing ISGylation via two deubiquitinases (USP16/USP18). Collectively, these findings not only establish DDX6 as a crucial host factor with potent antiviral activity but also enrich our understanding of host-virus interaction networks.</p>","PeriodicalId":20459,"journal":{"name":"Poultry Science","volume":"105 4","pages":"106529"},"PeriodicalIF":4.2,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12887166/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146113998","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-28DOI: 10.1016/j.psj.2026.106527
Manhong Wang , Xin Ye , Chuan-Yu Hsu , Hailey Fugate , Xue Zhang , Pratima Acharya Adhikari , Peixin Fan , Katie Elliott , Ken Macklin , Li Zhang
Metagenomic analyses have significantly advanced our understanding of microbial composition in the poultry gut. However, many microbes identified through metagenomic studies remain uncultured, largely due to the lack of understanding of their cultivation conditions, which hinders efforts to explore their functional roles in gut health and metabolism. In this study, we performed culturomics, a culture-dependent approach that combines diverse culture conditions with high-throughput 16S rRNA gene sequencing, to comprehensively assess the cultivability of chicken cecal microbiota and provide guidance for isolating target species of interest. Microbial profiling was performed using both culture-dependent (CD) and culture-independent (CI) approaches. For CI, genomic DNA (gDNA) was directly extracted from six broiler chicken cecal samples and subjected to full-length 16S rRNA gene sequencing. For CD, the same samples were cultured under 28 conditions, yielding 161 colony mixtures for sequencing. Based on diversity profiles of the colony mixtures, 10 conditions were selected for single-colony isolation and analysis. Results showed that CD and CI approaches identified 350 and 502 bacterial species, respectively, with 160 species detected by both methods. The dominant species recovered by the CD approach,including Escherichia coli, Proteus mirabilis, Limosilactobacillus reuteri, Enterococcus faecalis, and Ligilactobacillus salivarius, were detected at much lower abundances in the CI analysis, highlighting the capacity of culturomics to enrich and recover minority taxa that are often poorly detected by CI apparoach. Cultivation profiling showed that MRS selectively enriched Limosilactobacillus and Ligilactobacillus as well as Lactobacillus, whereas CNAB and MSA enriched Enterococcus and Bacillus, respectively. Community diversity and structure were significantly influenced by culture conditions (P < 0.01), with medium as the primary factor and air condition as a secondary factor. Subsequent single-colony analysis from 10 selected culture conditions identified 150 single-species isolates belonging to 14 distinct bacterial species. This study provides foundational insight into the cultivability of chicken cecal microbiota, facilitating future research to isolate specific strains and characterize their roles in poultry health and nutrition.
{"title":"Application of culturomics to explore the cultivable microbiota and enable targeted bacterial isolation from the ceca of broiler chickens","authors":"Manhong Wang , Xin Ye , Chuan-Yu Hsu , Hailey Fugate , Xue Zhang , Pratima Acharya Adhikari , Peixin Fan , Katie Elliott , Ken Macklin , Li Zhang","doi":"10.1016/j.psj.2026.106527","DOIUrl":"10.1016/j.psj.2026.106527","url":null,"abstract":"<div><div>Metagenomic analyses have significantly advanced our understanding of microbial composition in the poultry gut. However, many microbes identified through metagenomic studies remain uncultured, largely due to the lack of understanding of their cultivation conditions, which hinders efforts to explore their functional roles in gut health and metabolism. In this study, we performed culturomics, a culture-dependent approach that combines diverse culture conditions with high-throughput <em>16S rRNA</em> gene sequencing, to comprehensively assess the cultivability of chicken cecal microbiota and provide guidance for isolating target species of interest. Microbial profiling was performed using both culture-dependent (<strong>CD</strong>) and culture-independent (<strong>CI</strong>) approaches. For CI, genomic DNA (<strong>gDNA</strong>) was directly extracted from six broiler chicken cecal samples and subjected to full-length <em>16S rRNA</em> gene sequencing. For CD, the same samples were cultured under 28 conditions, yielding 161 colony mixtures for sequencing. Based on diversity profiles of the colony mixtures, 10 conditions were selected for single-colony isolation and analysis. Results showed that CD and CI approaches identified 350 and 502 bacterial species, respectively, with 160 species detected by both methods. The dominant species recovered by the CD approach,including <em>Escherichia coli, Proteus mirabilis, Limosilactobacillus reuteri, Enterococcus faecalis</em>, and <em>Ligilactobacillus salivarius</em>, were detected at much lower abundances in the CI analysis, highlighting the capacity of culturomics to enrich and recover minority taxa that are often poorly detected by CI apparoach. Cultivation profiling showed that MRS selectively enriched <em>Limosilactobacillus</em> and <em>Ligilactobacillus</em> as well as <em>Lactobacillus</em>, whereas CNAB and MSA enriched <em>Enterococcus</em> and <em>Bacillus</em>, respectively. Community diversity and structure were significantly influenced by culture conditions (<em>P</em> < 0.01), with medium as the primary factor and air condition as a secondary factor. Subsequent single-colony analysis from 10 selected culture conditions identified 150 single-species isolates belonging to 14 distinct bacterial species. This study provides foundational insight into the cultivability of chicken cecal microbiota, facilitating future research to isolate specific strains and characterize their roles in poultry health and nutrition.</div></div>","PeriodicalId":20459,"journal":{"name":"Poultry Science","volume":"105 4","pages":"Article 106527"},"PeriodicalIF":4.2,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146080500","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-28DOI: 10.1016/j.psj.2026.106542
Yuanhang Wei, Chimei Liao, Yixuan Wang, Can Cui, Yao Zhang, Zhuanjian Li, Congjiao Sun, Qing Zhu, Huadong Yin, Shunshun Han
Reproductive senescence in laying hens, characterized by a progressive decline in egg production, represents a major challenge for the poultry industry. Although microRNAs (miRNAs) are recognized as important regulators of aging, their specific roles and mechanisms in ovarian aging of hens remain largely unclear. This study was designed to comprehensively analyze miRNA expression patterns during ovarian aging in laying hens. The objectives of this study were to identify key functional miRNAs and to elucidate their molecular regulatory mechanisms. Specifically, this study evaluated ovarian senescence in hens at 350, 500 and 700 days of age, observing a decline in egg production, increased follicular atresia, and p53 upregulation. miRNA sequencing analysis identified 44 differentially expressed miRNAs (DEMs), among which gga-let-7i exhibited the highest abundance and showed progressive upregulation during aging. Functional assays revealed that gga-let-7i induces cell cycle arrest and promotes cellular senescence in ovarian follicle granulosa cells (GCs). Mechanistically, collagen type I alpha 2 chain (COL1A2) was confirmed as a direct target of gga-let-7i, and it has been demonstrated that gga-let-7i accelerates senescence by inhibiting the COL1A2/PI3K/AKT/MDM2 pathway, resulting in p53 accumulation and the downstream cellular senescence signaling pathways activation. These results uncover a novel gga-let-7i/COL1A2 regulatory axis involved in ovarian aging and suggest potential targets for extending reproductive longevity in laying hens.
{"title":"miRNA profiling reveals that gga-let-7i/COL1A2 axis induces cell cycle arrest and triggers cellular senescence to accelerate ovarian aging in laying hens by suppressing the PI3K/AKT/MDM2 pathway.","authors":"Yuanhang Wei, Chimei Liao, Yixuan Wang, Can Cui, Yao Zhang, Zhuanjian Li, Congjiao Sun, Qing Zhu, Huadong Yin, Shunshun Han","doi":"10.1016/j.psj.2026.106542","DOIUrl":"https://doi.org/10.1016/j.psj.2026.106542","url":null,"abstract":"<p><p>Reproductive senescence in laying hens, characterized by a progressive decline in egg production, represents a major challenge for the poultry industry. Although microRNAs (miRNAs) are recognized as important regulators of aging, their specific roles and mechanisms in ovarian aging of hens remain largely unclear. This study was designed to comprehensively analyze miRNA expression patterns during ovarian aging in laying hens. The objectives of this study were to identify key functional miRNAs and to elucidate their molecular regulatory mechanisms. Specifically, this study evaluated ovarian senescence in hens at 350, 500 and 700 days of age, observing a decline in egg production, increased follicular atresia, and p53 upregulation. miRNA sequencing analysis identified 44 differentially expressed miRNAs (DEMs), among which gga-let-7i exhibited the highest abundance and showed progressive upregulation during aging. Functional assays revealed that gga-let-7i induces cell cycle arrest and promotes cellular senescence in ovarian follicle granulosa cells (GCs). Mechanistically, collagen type I alpha 2 chain (COL1A2) was confirmed as a direct target of gga-let-7i, and it has been demonstrated that gga-let-7i accelerates senescence by inhibiting the COL1A2/PI3K/AKT/MDM2 pathway, resulting in p53 accumulation and the downstream cellular senescence signaling pathways activation. These results uncover a novel gga-let-7i/COL1A2 regulatory axis involved in ovarian aging and suggest potential targets for extending reproductive longevity in laying hens.</p>","PeriodicalId":20459,"journal":{"name":"Poultry Science","volume":"105 4","pages":"106542"},"PeriodicalIF":4.2,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146137684","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-28DOI: 10.1016/j.psj.2026.106535
Kaiyue Qin , Lingdan Yin , Huanrong Zhang
Since 2016, a novel astrovirus identified as goose astrovirus (GoAstV), is classified into genotypes 1 and 2 (GoAstV-1 and GoAstV-2). GoAstV-2 has caused a severe outbreak of visceral gout in goslings characterized by extensive visceral urate deposition and kidney swelling, resulting in substantial financial losses to the goose farming industry in China. The GoAstV-2 VP27 protein encoded by ORF2 contains neutralizing antigenic epitopes, thus representing a key candidate for the development of diagnostic reagents and epitope vaccines. This study aimed to prepare the monoclonal antibody (mAb) against GoAstV-2 VP27 and identify its epitope. The GoAstV-2 VP27 protein was expressed and purified using a prokaryotic expression system, followed by immunization of BALB/c mice. We employed hybridoma technology to generate a stable monoclonal antibody-secreting cell line targeting GoAstV-2 VP27, which was named 11-39B and characterized as IgG2b with kappa light chain. Furthermore, the mAb 11-39B specifically bound to GoAstV-2 as confirmed by Western blotting (WB), immunofluorescence assay (IFA) and immunohistochemistry (IHC), and potently neutralized GoAstV-2 infection in vivo in a dose-dependent manner. For epitope mapping, sequential truncations of the GoAstV-2 VP27 protein were constructed by eukaryotic expression and tested by WB. The results indicated that peptide 661SLKTS665 was the minimal epitope recognized by mAb 11-39B. Homology and structural analyses demonstrated that the epitope was situated on the surface of the VP27 protein and exhibited high conservation among GoAstV-2 strains but exhibited significant differences in the GoAstV-1 serotype. Our study contributes to a better understanding of the GoAstV-2 VP27 antigenic region and provides a basis for establishing epitope-based GoAstV-2 diagnostic methods and vaccine development.
{"title":"Development of monoclonal antibodies for GoAstV-2 VP27 protein and precise mapping of linear antigenic epitopes","authors":"Kaiyue Qin , Lingdan Yin , Huanrong Zhang","doi":"10.1016/j.psj.2026.106535","DOIUrl":"10.1016/j.psj.2026.106535","url":null,"abstract":"<div><div>Since 2016, a novel astrovirus identified as goose astrovirus (GoAstV), is classified into genotypes 1 and 2 (GoAstV-1 and GoAstV-2). GoAstV-2 has caused a severe outbreak of visceral gout in goslings characterized by extensive visceral urate deposition and kidney swelling, resulting in substantial financial losses to the goose farming industry in China. The GoAstV-2 VP27 protein encoded by ORF2 contains neutralizing antigenic epitopes, thus representing a key candidate for the development of diagnostic reagents and epitope vaccines. This study aimed to prepare the monoclonal antibody (mAb) against GoAstV-2 VP27 and identify its epitope. The GoAstV-2 VP27 protein was expressed and purified using a prokaryotic expression system, followed by immunization of BALB/c mice. We employed hybridoma technology to generate a stable monoclonal antibody-secreting cell line targeting GoAstV-2 VP27, which was named 11-39B and characterized as IgG2b with kappa light chain. Furthermore, the mAb 11-39B specifically bound to GoAstV-2 as confirmed by Western blotting (WB), immunofluorescence assay (IFA) and immunohistochemistry (IHC), and potently neutralized GoAstV-2 infection <em>in vivo</em> in a dose-dependent manner. For epitope mapping, sequential truncations of the GoAstV-2 VP27 protein were constructed by eukaryotic expression and tested by WB. The results indicated that peptide <sup>661</sup>SLKTS<sup>665</sup> was the minimal epitope recognized by mAb 11-39B. Homology and structural analyses demonstrated that the epitope was situated on the surface of the VP27 protein and exhibited high conservation among GoAstV-2 strains but exhibited significant differences in the GoAstV-1 serotype. Our study contributes to a better understanding of the GoAstV-2 VP27 antigenic region and provides a basis for establishing epitope-based GoAstV-2 diagnostic methods and vaccine development.</div></div>","PeriodicalId":20459,"journal":{"name":"Poultry Science","volume":"105 4","pages":"Article 106535"},"PeriodicalIF":4.2,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146080938","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-28DOI: 10.1016/j.psj.2026.106526
Hongbo Zeng, Junjie Zhang, Fei Wang, Yang Wen, Jie Cai, Yingping Xiao, Bindan Chen, Hua Yang, Wentao Lyu
Salmonella infections pose a significant threat to waterfowl. Avian β-defensin 10 (AvBD10), a key molecule of host innate immunity, exhibits broad-spectrum antibacterial activity and immune regulatory function. However, research on the tissue-specific expression and antibacterial activity against Salmonella of AvBD10 in waterfowl remains limited. In this study, we first revealed the expression characteristics of AvBD10 in the tissues of ducks at different ages using in vivo experiments. Subsequently, we elucidated the interaction between AvBD10 and intestinal microorganisms, such as Salmonella, by combining 16S rDNA sequencing analysis of the microbiota in the cecum. Finally, we verified the antibacterial effect of AvBD10 on Salmonella using in vitro experiments. The results revealed that the expression of AvBD10 displayed notable (P<0.05) tissue-specific patterns, exhibiting variations associated with the developmental stage in ducks. In particular, AvBD10 was prominently expressed in the cecum during the initial brooding phase, with consistent modulation observed throughout maturation. In addition, AvBD10 expression shows a significant negative correlation (R=-0.460; 95% confidence intervals: -0.632 to -0.248; P<0.05) with the relative abundance of Salmonella in the cecum. Furthermore, AvBD10 exhibited a minimum inhibitory concentration of 62.5 μg/mL against Salmonella enterica and exerted its antibacterial properties by electrostatically binding to the Salmonella cell membrane via positively charged amino acid residues, leading to disruption of the membrane structure. This study provides a significant theoretical foundation for the green and healthy breeding of livestock and poultry.
{"title":"Duck β-defensin 10 inhibits Salmonella enterica by disrupting bacterial membrane integrity and its association with gut microbiota dynamics.","authors":"Hongbo Zeng, Junjie Zhang, Fei Wang, Yang Wen, Jie Cai, Yingping Xiao, Bindan Chen, Hua Yang, Wentao Lyu","doi":"10.1016/j.psj.2026.106526","DOIUrl":"10.1016/j.psj.2026.106526","url":null,"abstract":"<p><p>Salmonella infections pose a significant threat to waterfowl. Avian β-defensin 10 (AvBD10), a key molecule of host innate immunity, exhibits broad-spectrum antibacterial activity and immune regulatory function. However, research on the tissue-specific expression and antibacterial activity against Salmonella of AvBD10 in waterfowl remains limited. In this study, we first revealed the expression characteristics of AvBD10 in the tissues of ducks at different ages using in vivo experiments. Subsequently, we elucidated the interaction between AvBD10 and intestinal microorganisms, such as Salmonella, by combining 16S rDNA sequencing analysis of the microbiota in the cecum. Finally, we verified the antibacterial effect of AvBD10 on Salmonella using in vitro experiments. The results revealed that the expression of AvBD10 displayed notable (P<0.05) tissue-specific patterns, exhibiting variations associated with the developmental stage in ducks. In particular, AvBD10 was prominently expressed in the cecum during the initial brooding phase, with consistent modulation observed throughout maturation. In addition, AvBD10 expression shows a significant negative correlation (R=-0.460; 95% confidence intervals: -0.632 to -0.248; P<0.05) with the relative abundance of Salmonella in the cecum. Furthermore, AvBD10 exhibited a minimum inhibitory concentration of 62.5 μg/mL against Salmonella enterica and exerted its antibacterial properties by electrostatically binding to the Salmonella cell membrane via positively charged amino acid residues, leading to disruption of the membrane structure. This study provides a significant theoretical foundation for the green and healthy breeding of livestock and poultry.</p>","PeriodicalId":20459,"journal":{"name":"Poultry Science","volume":"105 4","pages":"106526"},"PeriodicalIF":4.2,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12882717/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146100521","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Background: Pasteurella multocida, the cause of fowl cholera, poses a significant economic threat to poultry. The escalation of antimicrobial resistance (AMR) in this pathogen, driven by drug misuse, is a serious concern. Crucially, a comprehensive understanding of AMR patterns, molecular epidemiology, and virulence in Chinese avian strains is still lacking. This study aimed to elucidate the phenotypic and genomic characteristics of P. multocida strains isolated from avian in Zhejiang Province, China.
Result: Twenty avian-origin P. multocida strains isolated in Zhejiang Province were subjected to antimicrobial susceptibility testing, whole-genome sequencing, and virulence assessment. Resistance was most frequent to florfenicol (65%) and tetracycline (50%). Multi-Locus Sequence Typing analysis revealed that ST471 and its closely related variant ST129 collectively constituted the dominant clonal group. A total of 13 resistance genes belonging to 5 major classes were identified, with floR (75%), sul2 (50%), and tet (B) (50%) exhibiting the highest prevalence. In the Galleria mellonella model challenged with 1.5 × 10⁷ CFU/larva, ST471 strains exhibited a slower killing rate compared to ST129 strains, while the ST7 strain demonstrated low virulence.
Conclusion: This study demonstrates the increasing prevalence of avian P. multocida in China, specifically the sustained circulation of ST129 and the recent emergence of ST471, particularly in ducks. These findings underscore the urgency of continuous monitoring of strain dissemination and the evolution of multidrug resistance, providing a scientific basis for precise and rational antimicrobials use in farms and for blocking the spread of potentially high-risk strains.
{"title":"Antimicrobial resistance and genomic characteristics of avian Pasteurella multocida.","authors":"Fengcheng Miao, Bing Dai, Zhiyu Li, Kaikai Lv, Junxing Li, Shuangmao Li, Shuaijie Song, Huafeng Jian, Xiaoqian Long, Yao Shen, Yinghui Wei, Hua Yang, Jiangang Ma","doi":"10.1016/j.psj.2026.106524","DOIUrl":"https://doi.org/10.1016/j.psj.2026.106524","url":null,"abstract":"<p><strong>Background: </strong>Pasteurella multocida, the cause of fowl cholera, poses a significant economic threat to poultry. The escalation of antimicrobial resistance (AMR) in this pathogen, driven by drug misuse, is a serious concern. Crucially, a comprehensive understanding of AMR patterns, molecular epidemiology, and virulence in Chinese avian strains is still lacking. This study aimed to elucidate the phenotypic and genomic characteristics of P. multocida strains isolated from avian in Zhejiang Province, China.</p><p><strong>Result: </strong>Twenty avian-origin P. multocida strains isolated in Zhejiang Province were subjected to antimicrobial susceptibility testing, whole-genome sequencing, and virulence assessment. Resistance was most frequent to florfenicol (65%) and tetracycline (50%). Multi-Locus Sequence Typing analysis revealed that ST471 and its closely related variant ST129 collectively constituted the dominant clonal group. A total of 13 resistance genes belonging to 5 major classes were identified, with floR (75%), sul2 (50%), and tet (B) (50%) exhibiting the highest prevalence. In the Galleria mellonella model challenged with 1.5 × 10⁷ CFU/larva, ST471 strains exhibited a slower killing rate compared to ST129 strains, while the ST7 strain demonstrated low virulence.</p><p><strong>Conclusion: </strong>This study demonstrates the increasing prevalence of avian P. multocida in China, specifically the sustained circulation of ST129 and the recent emergence of ST471, particularly in ducks. These findings underscore the urgency of continuous monitoring of strain dissemination and the evolution of multidrug resistance, providing a scientific basis for precise and rational antimicrobials use in farms and for blocking the spread of potentially high-risk strains.</p>","PeriodicalId":20459,"journal":{"name":"Poultry Science","volume":"105 4","pages":"106524"},"PeriodicalIF":4.2,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146119744","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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