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22 pages, 1869 KB  
Review
Curcumin as a Green Antibiotic Substitute: Mechanisms and Applications in Poultry Production and Health Promotion
by Xiaopeng Tang, Baoshan Zhang, Jiayuan Yang, Youyuan Xie and Kangning Xiong
Animals 2026, 16(8), 1242; https://doi.org/10.3390/ani16081242 - 17 Apr 2026
Viewed by 562
Abstract
Against the backdrop of the full implementation of “antibiotic ban” and “zinc restriction” policies in livestock and poultry breeding, and the growing consumer demand for safe livestock and poultry products, the development of natural and efficient green feed additives has become crucial for [...] Read more.
Against the backdrop of the full implementation of “antibiotic ban” and “zinc restriction” policies in livestock and poultry breeding, and the growing consumer demand for safe livestock and poultry products, the development of natural and efficient green feed additives has become crucial for the sustainable development of the animal husbandry industry. Curcumin, a natural polyphenolic compound extracted from the rhizome of Curcuma longa L., has attracted extensive attention in poultry production due to its various biological activities and safety. This paper thoroughly reviews the chemical structure and physicochemical properties of curcumin, and elaborates on its core molecular mechanisms of action, which mainly involve the regulation of nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE), nuclear factor-κB (NF-κB), peroxisome proliferator-activated receptor γ (PPAR-γ), and mitogen-activated protein kinase (MAPK) pathways to exert antioxidant, anti-inflammatory, antibacterial, immunomodulatory and lipid metabolism regulatory effects. It further clarifies the practical application value of curcumin in major poultry species including broilers, laying hens, ducks and quails, showing that curcumin can significantly improve poultry production performance, optimize meat and egg quality, protect intestinal health, and enhance the ability of poultry to resist stress and diseases. Meanwhile, the review notes curcumin’s current application limitations (low bioavailability, poor stability, unclear standardized dosage, and high industrialization cost) and proposes targeted future research directions to address these issues. In conclusion, curcumin is a promising green feed additive alternative to antibiotics, and its large-scale and standardized application in poultry production will effectively promote the green, healthy and sustainable development of the poultry industry. Full article
(This article belongs to the Section Poultry)
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23 pages, 4180 KB  
Article
Efficacy and Limitations of an Improved Vaccine Derived from an Updated Vaccine Strain Against H5 High Pathogenicity Avian Influenza
by Bao Linh Nguyen, Norikazu Isoda, Yik Lim Hew, Loc Tan Huynh, Kien Trung Le, Yo Shimazu, Daiki Kobayashi, Dang Hoang Nguyen, Tho Dang Nguyen, Duc-Huy Chu, Diep Thi Nguyen, Koki Takeichi, Yuto Nanba, Takahiro Hiono, Takashi Sasaki and Yoshihiro Sakoda
Vaccines 2026, 14(4), 291; https://doi.org/10.3390/vaccines14040291 - 24 Mar 2026
Viewed by 678
Abstract
Background/Objectives: Biosecurity and stamping out are key control measures against H5 high pathogenicity avian influenza (HPAI) outbreaks. Vaccination in poultry is an additional tool to reduce disease risk and facilitate timely containment. This study aimed to establish a candidate vaccine strain against H5 [...] Read more.
Background/Objectives: Biosecurity and stamping out are key control measures against H5 high pathogenicity avian influenza (HPAI) outbreaks. Vaccination in poultry is an additional tool to reduce disease risk and facilitate timely containment. This study aimed to establish a candidate vaccine strain against H5 HPAI in Asia and validate its protective efficacy. Methods: Based on genetic and antigenic analyses, a representative HPAI virus, A/duck/Vietnam/HU16-DD3/2023 (H5N1), collected in northern Vietnam, was selected to generate a candidate vaccine strain, rgPR8/VN23HA∆KRRK-NA (rgPR8/VN23; H5N1), using reverse genetics, followed by formulation of an inactivated oil-adjuvanted vaccine. Vaccine efficacy was evaluated by measuring humoral antibody responses after intramuscular vaccination and by assessing mortality and virus recovery following intranasal challenge with a clade 2.3.4.4b virus, A/Ezo red fox/Hokkaido/1/2022 (H5N1). Results were compared with those obtained using an antigenically homologous vaccine to the challenge strain and a Japanese stockpiled vaccine. Results: All vaccinated juvenile chickens developed sufficient immunity to survive the challenge at 21 days post-vaccination. The rgPR8/VN23 (H5N1) and homologous vaccines markedly reduced virus recovery, suggesting near-sterile protection, whereas low-titer viruses were transiently detected in chickens vaccinated with the stockpiled vaccine. The rgPR8/VN23 (H5N1) vaccine conferred clinical protection in juvenile chickens as early as 8 days post-vaccination. A single dose of the rgPR8/VN23 (H5N1) vaccine provided incomplete protection in laying hens, whereas a double-volume regimen improved protective efficacy. Conclusions: The rgPR8/VN23 (H5N1) vaccine conferred strong immunity to juvenile chickens; however, a refined vaccination strategy may be required to achieve complete protection in laying hens. Full article
(This article belongs to the Special Issue Immunity to Influenza Viruses and Vaccines)
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18 pages, 2943 KB  
Article
Transcriptomic Profiling Identifies Key Genes and ERBB Signaling Pathway Associated with Aggressive Behavior in Muscovy Ducks (Cairina moschata)
by Ai Liu, Xuping Wang, Xuan Zhou, Biqiong Yao, Jinjin Zhu, Yifu Rao, Fuyou Liao, Bingnong Yao, Surintorn Boonanuntan and Shenglin Yang
Animals 2026, 16(6), 951; https://doi.org/10.3390/ani16060951 - 18 Mar 2026
Viewed by 399
Abstract
Aggressive behavior in Muscovy ducks (Cairna moschata) has become a predominant concern in intensive farming systems, leading to reduced animal welfare and production losses. To unravel the molecular mechanisms underlying this behavior, transcriptomic profiling was performed on the hypothalamus, a key regulatory hub [...] Read more.
Aggressive behavior in Muscovy ducks (Cairna moschata) has become a predominant concern in intensive farming systems, leading to reduced animal welfare and production losses. To unravel the molecular mechanisms underlying this behavior, transcriptomic profiling was performed on the hypothalamus, a key regulatory hub for aggressive responses. A total of 120 healthy 60-day-old female Muscovy ducks were continuously monitored for 24 h/day over one month using Media Recorder 2.0 software. Based on instantaneous and continuous behavioral observations, the ducks were categorized into three groups: aggressor (Experimental group I, actively attacking conspecifics), victim (Experimental group II, receiving aggression), and non-aggressive (Control group, no aggressive interactions). Hypothalamic tissues were collected from each group (n = 4 per group) for Illumina HiSeq 2000 high-throughput transcriptome sequencing. Functional annotation and enrichment analysis of differentially expressed genes (DEGs) were performed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases, followed by quantitative real-time PCR (qRT-PCR) validation. GO analysis identified 626 DEGs in the aggressor group and 649 DEGs in the victim group compared to the control group, with 26 DEGs directly involved in aggressive behavior regulation. Integration of GO and KEGG annotations revealed 69 candidate genes associated with aggressive behavior, enriched in two GO terms (behavior [GO:0007610] and sensory perception of pain [GO:0019233]) and the ERBB signaling pathway (map04012). qRT-PCR validation of 14 randomly selected candidate genes (e.g., NPY, ERBB4, MAPK9, PRDM12) confirmed that their expression patterns were consistent with transcriptomic data, verifying the reliability of the sequencing results. These findings provide novel insights into the molecular genetic basis of aggressive behavior in Muscovy ducks and lay a foundation for developing targeted strategies to mitigate aggression in intensive farming systems. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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9 pages, 436 KB  
Article
Assessment of Compliance with Animal Welfare Requirements Across Poultry Species and Production Categories
by Eva Justova, Vladimir Vecerek, Zbynek Semerad, Marijana Vucinic and Eva Voslarova
Animals 2026, 16(5), 834; https://doi.org/10.3390/ani16050834 - 7 Mar 2026
Viewed by 514
Abstract
Animal welfare is a key component of sustainable poultry production and is routinely monitored through official veterinary inspections. The aim of this study was to determine the level of welfare compliance among different poultry species and production categories, to compare compliance levels across [...] Read more.
Animal welfare is a key component of sustainable poultry production and is routinely monitored through official veterinary inspections. The aim of this study was to determine the level of welfare compliance among different poultry species and production categories, to compare compliance levels across these groups, and to assess long-term trends using official inspection data. The study was based on the results of supervisory inspections conducted by veterinary inspectors in poultry farms in the Czech Republic between 2016 and 2024. Welfare compliance was evaluated in laying hens, broiler chickens, turkeys, ducks, and geese using a standardized system of welfare assessment checkpoints applied during official controls. Inspections were classified as compliant or non-compliant based on the presence or absence of deficiencies, and overall compliance levels were calculated as the proportion of animals kept in farms with compliant inspections. Across the entire study period, the proportion of poultry kept in farms with compliant inspections ranged from 82.8% to 98.4%, with the highest compliance level observed in turkeys, followed by ducks and broiler chickens, while the lowest compliance level was recorded in geese. Differences among poultry species and categories were statistically significant (p < 0.05). Comparison of two time periods (2016–2018 and 2022–2024) revealed significant improvements in compliance for broiler chickens, ducks, and geese, whereas significant declines were observed for laying hens and turkeys (p < 0.001). These results demonstrate clear differences in welfare compliance among poultry species and categories and indicate that compliance trends over time are not uniform across the poultry sector. Official veterinary inspection data provide a valuable tool for large-scale assessment of welfare compliance and for identifying poultry sectors that may benefit from targeted welfare improvement measures. Full article
(This article belongs to the Section Poultry)
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15 pages, 4226 KB  
Article
Transcriptomic Analysis Reveals Sex-Biased Gene Expression in Duck Turbinate Tissue
by Kangling Li, Kexin Wu, Qinglian Li, Xintong Yu, Ruolan Li, Mao Chen, Xu Han, Hehe Liu and Anqi Huang
Animals 2026, 16(5), 714; https://doi.org/10.3390/ani16050714 - 25 Feb 2026
Viewed by 489
Abstract
Olfaction is crucial for ducks, influencing essential behaviors such as foraging and mating. However, the molecular basis of sex-associated variation in duck olfactory tissues remains poorly understood. Here, we performed bulk RNA-seq on turbinate tissue from male and female Tianfu Nonghua Mottled Ducks [...] Read more.
Olfaction is crucial for ducks, influencing essential behaviors such as foraging and mating. However, the molecular basis of sex-associated variation in duck olfactory tissues remains poorly understood. Here, we performed bulk RNA-seq on turbinate tissue from male and female Tianfu Nonghua Mottled Ducks (Anas platyrhynchos domesticus Linnaeus, 1758; Anatidae) to characterize sex-biased transcriptional programs. Our results suggest strong global transcriptomic separation between males and females, with 1906 differentially expressed genes (DEGs) identified. These DEGs were enriched in pathways related to neuronal signaling, cell adhesion, and extracellular matrix organization, suggesting coordinated sex-associated differences in signaling and tissue-organization programs. While olfactory receptor (OR) and trace amine-associated receptor (TAAR) genes showed limited sex-biased expression in bulk tissue, two neuromodulatory GPCRs, TACR2 and DRD4, were prioritized as hub genes within sex-biased co-expression networks. Notably, both genes also showed relatively high expression in turbinate tissue and neuroendocrine centers in an integrated multi-tissue transcriptomic dataset, nominating them as candidate targets for future functional and cell-type-resolved investigations. Overall, our study provides a descriptive molecular profile of sex-biased transcription in duck turbinate tissue, laying a foundation for follow-up studies and potential applications in poultry breeding and management. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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15 pages, 2902 KB  
Article
Duck IFNγ Restricts Duck Tembusu Virus Replication by Disrupting Viral RNA Synthesis/Translation
by Juan Huang, Xinyue Li, Yuxin Lu, Shun Chen, Bin Tian, Renyong Jia and Anchun Cheng
Vet. Sci. 2026, 13(2), 178; https://doi.org/10.3390/vetsci13020178 - 11 Feb 2026
Viewed by 841
Abstract
Duck Tembusu virus (DTMUV), an emerging Flavivirus, is a major avian pathogen that imposes enormous economic losses on the global duck industry, necessitating urgent development of effective countermeasures. Although Interferon-gamma (IFNγ) is a crucial broad-spectrum antiviral cytokine, its role against DTMUV infection [...] Read more.
Duck Tembusu virus (DTMUV), an emerging Flavivirus, is a major avian pathogen that imposes enormous economic losses on the global duck industry, necessitating urgent development of effective countermeasures. Although Interferon-gamma (IFNγ) is a crucial broad-spectrum antiviral cytokine, its role against DTMUV infection remains mechanistically undefined. In this study, we first demonstrated that DTMUV induced duck IFNγ (duIFNγ) production in immune and non-immune cells. Importantly, duIFNγ exhibited a dual anti-DTMUV function in vitro: it not only prevented viral replication but also displayed the capacity to clear existing virus from infected cells. Mechanistically, cycloheximide (CHX) experiments confirmed that duIFNγ exerts its antiviral effect by disrupting the viral RNA synthesis/translation phase. Furthermore, transcriptomic profiling (RNA-seq) precisely revealed that duIFNγ restricts DTMUV replication by activating multiple host defense pathways, notably Programmed Cell Death (e.g., Caspase signaling) and the RIG-I-like Receptor (RLR) signaling pathways. Collectively, these findings provide critical insights into the function and mechanism of duIFNγ in combating DTMUV in vitro, laying a robust theoretical foundation for exploring duIFNγ or its induced effectors as novel therapeutics for DTMUV infection. Full article
(This article belongs to the Section Veterinary Biomedical Sciences)
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16 pages, 1452 KB  
Article
Molecular Characterization of Highly Pathogenic Avian Influenza H5N1 Viruses Circulating in Bulgaria During 2024–2025: Evidence for Hidden Circulation and Zoonotic Risk Markers
by Gabriela Goujgoulova, Georgi Stoimenov and Koycho Koev
Int. J. Mol. Sci. 2026, 27(4), 1711; https://doi.org/10.3390/ijms27041711 - 10 Feb 2026
Viewed by 678
Abstract
The highly pathogenic avian influenza virus HPAI A(H5N1) genotype AF was detected in southern Europe during the 2021/2022 season and spread widely. It emerged in Bulgaria in 2022/2023, mainly affecting mallard ducks. The DA genotype of the virus was detected in a diverse [...] Read more.
The highly pathogenic avian influenza virus HPAI A(H5N1) genotype AF was detected in southern Europe during the 2021/2022 season and spread widely. It emerged in Bulgaria in 2022/2023, mainly affecting mallard ducks. The DA genotype of the virus was detected in a diverse group of birds, including wild birds, zoo birds, and domestic poultry, across a wide area of eastern and southern Europe in 2023. In Bulgaria, following its introduction in 2023, the DA genotype became the predominant virus in laying hens. During 2024–2025, DA spread throughout the country, displacing AF from mallard flocks. The predominant subtype in Europe in 2025 was H5N1 genotype DI.2. This genotype became dominant after December 2024, accounting for over 90% of viruses within the EA-2024-DI genotype lineage, and has been detected in a wide range of bird species. In Bulgaria, DI.2 was identified in only one outbreak in a flock of laying hens in autumn 2024 and in a single case involving a western marsh harrier (Circus aeruginosus) in early 2025. These observations are consistent with a pattern of putative hidden circulation of avian influenza virus in duck farms in Bulgaria, potentially establishing a cycle of continuous circulation of the same viral subtype. In this study, we analysed viruses originating from Bulgaria, with a particular focus on EA-2024-DI genotype DI.2, and examined mutations related to host cell receptor binding, host specificity shifts, ligand binding, antibody recognition sites, viral oligomerization interfaces, and other functional regions. Some of these mutations have been associated with antigenic drift, immune escape, and virulence. Importantly, several are linked to changes in host specificity, a critical step in the potential transition of avian influenza viruses to humans. Consequently, such mutations represent key factors in the spread of highly pathogenic avian influenza and may pose a pandemic risk. Full article
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20 pages, 4455 KB  
Article
A Comparative Analysis of Transcriptome-Wide Differential Gene Expression and Alternative Polyadenylation in the Ovaries of Meat Ducks and Laying Ducks
by Sike Wang, Yaomei Wang, Shiwei Li, Chao Jia, Debing Yu and Weiling Huang
Animals 2026, 16(2), 313; https://doi.org/10.3390/ani16020313 - 20 Jan 2026
Viewed by 539
Abstract
Significant differences in reproductive performance exist between meat-type ducks (e.g., Qiangying Duck, QD) and laying-type ducks (e.g., Shaoxing Duck, SD). The molecular mechanisms underlying these differences, particularly concerning ovarian development and function, remain incompletely understood. This study aimed to comprehensively characterize the ovarian [...] Read more.
Significant differences in reproductive performance exist between meat-type ducks (e.g., Qiangying Duck, QD) and laying-type ducks (e.g., Shaoxing Duck, SD). The molecular mechanisms underlying these differences, particularly concerning ovarian development and function, remain incompletely understood. This study aimed to comprehensively characterize the ovarian transcriptomes of these two duck types, focusing on differential gene expression and post-transcriptional regulatory events. We performed an integrated full-length transcriptome analysis of ovarian tissues from these two breeds using PacBio SMRT and Illumina sequencing. Bioinformatic analyses, including functional annotation, differential expression analysis, and the identification of APA events, were used. We discovered substantial breed-specific differences in alternative polyadenylation (APA), with SD ducks exhibiting significant 3′UTR shortening in 3799 genes and 3′UTR lengthening in 1626 genes compared to QD. The integrated analysis of differential gene expression and APA events highlighted key genes related to steroid hormone synthesis (HMGCS1, DHCR24), lipid metabolism (SCD), signal transduction (HRAS), and antioxidant defense (SOD1). The functional enrichment implicated critical pathways such as mitochondrial energy metabolism, oxidative phosphorylation, and fatty acid degradation. Our study provides a comprehensive atlas of post-transcriptional regulation in the duck ovary and reveals APA as a crucial process of gene regulation. APA may contribute to the differential ovarian function and egg-laying capacity between meat and laying ducks, thus offering valuable targets for genetic selection. Full article
(This article belongs to the Special Issue Advances in Genetic Analysis of Important Traits in Poultry)
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16 pages, 3064 KB  
Article
Curcumin Mitigates Fumonisin B1-Induced Ovarian Toxicity in Peak-Laying Ducks via Hormone Metabolic Protection and Enhanced Reproductive Resilience
by Lihua Wang, Rui Liang, Qingyun Cao, Zhiwei Hou, Ali Mujtaba Shah, Qiuyi Deng, Xue Li, Jinze Li, Jiaqing Chen, Lukuyu A. Bernard, Muhammad Kashif Saleemi, Lin Yang and Wence Wang
Toxins 2026, 18(1), 34; https://doi.org/10.3390/toxins18010034 - 9 Jan 2026
Cited by 1 | Viewed by 715
Abstract
The objective of this study was to evaluate the protective effect of curcumin (Cur) on reproductive toxicity induced by fumonisin B1 (FB1) in laying ducks during the peak egg-laying period. A total of seventy-two 50-week-old Cherry Valley ducks were randomly [...] Read more.
The objective of this study was to evaluate the protective effect of curcumin (Cur) on reproductive toxicity induced by fumonisin B1 (FB1) in laying ducks during the peak egg-laying period. A total of seventy-two 50-week-old Cherry Valley ducks were randomly assigned to four groups: control, FB1 (30 mg/kg), Cur (200 mg/kg), and Cur + FB1 (200 mg/kg + 30 mg/kg). The experiment lasted for 35 days. Our results showed that cur supplementation effectively restored the reductions in final body weight (p = 0.005) and oviduct length (p = 0.020) induced by FB1 exposure. Residual FB1 concentrations in serum, liver, and ovaries were markedly increased in the FB1-treated group, while Cur significantly decreased the FB1 residual in duck liver (p < 0.05). Meanwhile, Cur supplementation markedly counteracted the FB1-induced reductions in serum total protein, albumin, triglycerides, and high-density lipoprotein induced by FB1 exposure. Cur supplementation effectively regulated FB1-induced oxidative stress, inflammation, and endocrine disruption. Specifically, Cur lowered FB1-induced malondialdehyde levels (p < 0.010), attenuated interleukin-1β increase (p = 0.083), and reversed the reduction in immunoglobulin G levels. FB increased the levels of hormones associated with duck reproduction, including estradiol, follicle-stimulating hormone, and luteinizing hormone; in contrast, curcumin supplementation decreased the levels of these hormones (p < 0.010). Histopathological analysis revealed that Cur significantly alleviated the inflammation and necrosis in the liver, kidneys, ovaries, and oviducts induced by FB1. In conclusion, dietary Cur supplementation effectively alleviated FB1-induced reproductive toxicity in laying ducks by enhancing antioxidant capacity, improving lipid metabolism, and restoring hormonal homeostasis. Full article
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15 pages, 2615 KB  
Article
5-HEPE Ameliorates Aging of Duck Ovarian Granulosa Cells by Targeting FOXM1 and Suppressing Oxidative Stress
by Yibo Zong, Jinyu Liu, Wenwu Xu, Tiantian Gu, Yong Tian, Li Chen, Tao Zeng and Lizhi Lu
Antioxidants 2025, 14(12), 1425; https://doi.org/10.3390/antiox14121425 - 27 Nov 2025
Cited by 1 | Viewed by 741
Abstract
Oxylipins are tightly linked to ovarian function. However, the roles and mechanisms of oxylipins in ovarian aging of laying ducks remain poorly understood. In this study, 72-week-old laying Jinyun ducks were categorized into high-laying (HL) and low-laying (LL) groups. Plasma and ovarian tissues [...] Read more.
Oxylipins are tightly linked to ovarian function. However, the roles and mechanisms of oxylipins in ovarian aging of laying ducks remain poorly understood. In this study, 72-week-old laying Jinyun ducks were categorized into high-laying (HL) and low-laying (LL) groups. Plasma and ovarian tissues were analyzed for antioxidant indices, transcriptomic, and targeted lipidomic. Results demonstrated that, compared to the LL group, the HL group ducks exhibited lower levels of MDA in both plasma and ovarian tissues, while exhibiting higher levels of SOD, GSH-Px, and T-AOC. Differentially expressed genes were primarily enriched in fatty acid oxidation and inflammation-related pathways, with FOXM1 identified as a pivotal gene involved in delaying ovarian aging. Furthermore, oxylipin profiles in plasma and ovary exhibit marked distinctions between the HL and LL group ducks. Notably, 5-hydroxy-eicosapentaenoic acid (5-HEPE) was substantially upregulated in both plasma and ovaries of the HL group ducks. Meanwhile, 5-HEPE significantly enhanced FOXM1 expression and mitigated oxidative stress in granulosa cells. Collectively, this study provides the first comprehensive analysis of oxylipin changes in the ovaries and plasma of ducks with differing laying performances at the late-laying stage. These findings offer novel insights into the prevention and alleviation of ovarian oxidative stress in ducks during this period. Full article
(This article belongs to the Section Health Outcomes of Antioxidants and Oxidative Stress)
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20 pages, 3706 KB  
Article
A Highly Immunogenic and Cross-Reactive Multi-Epitope Vaccine Candidate Against Duck Hepatitis A Virus: Immunoinformatics Design and Preliminary Experimental Validation
by Yuanhe Yang, Xiaodong Chen, Anguo Liu, Jinxin He, Yunhe Cao and Pingli He
Int. J. Mol. Sci. 2025, 26(22), 10958; https://doi.org/10.3390/ijms262210958 - 12 Nov 2025
Cited by 1 | Viewed by 1227
Abstract
Duck viral hepatitis (DVH), a highly contagious disease, is caused primarily by duck hepatitis A virus (DHAV). The viral genotypes exhibit significant diversity, creating a challenge as monovalent vaccines fail to provide cross-genotype protection in ducklings. This study aimed to design a multi-epitope [...] Read more.
Duck viral hepatitis (DVH), a highly contagious disease, is caused primarily by duck hepatitis A virus (DHAV). The viral genotypes exhibit significant diversity, creating a challenge as monovalent vaccines fail to provide cross-genotype protection in ducklings. This study aimed to design a multi-epitope peptide vaccine targeting different genotypes of DHAV. Using immunoinformatics approaches, we systematically identified key antigenic determinants, including linear B-cell epitopes, cytotoxic T-cell epitopes (CTL), and helper T-cell epitopes (HTL). Based on these, a novel vaccine candidate was developed. The vaccine construct was subjected to rigorous computational validation: (1) Molecular docking with Toll-like receptors (TLRs) predicted immune interaction potential. (2) Molecular dynamics simulations assessed complex stability. (3) In silico cloning ensured prokaryotic expression feasibility. Then, we conducted preliminary experimental validation for the actual effect of the vaccine candidate, including recombinant protein expression in E. coli, enzyme-linked immunosorbent assay (ELISA) quantification of humoral responses, and Western blot analysis of cross-reactivity. ELISA results demonstrated that the vaccine candidate could induce high-titer antibodies in immunized animals, with potency reaching up to 1:128,000, and the immune serum showed strong reactivity with recombinant VP proteins. Western blot analysis using duck sera confirmed epitope conservancy across genotypes. Collectively, the multi-epitope vaccine candidate developed in this study represents a highly promising broad-spectrum strategy against DHAV. The robust humoral immunity it elicits, coupled with its demonstrated cross-reactivity, constitutes compelling proof-of-concept, laying a solid foundation for advancing to subsequent challenge trials and translational applications. Full article
(This article belongs to the Special Issue Advances in Vaccine Immunology)
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22 pages, 5862 KB  
Article
Construction of a Duck Intestinal Organoid Culture System: From Crypt Isolation to Medium Optimization
by Rui Tang, Xiang Luo, Li Zhang, Zhenhua Liang, Yan Wu, Jingbo Liu, Jinsong Pi and Hao Zhang
Animals 2025, 15(21), 3145; https://doi.org/10.3390/ani15213145 - 29 Oct 2025
Viewed by 1153
Abstract
Intestinal organoids possess self-organizing capacity and recapitulate essential features of intestinal architecture and function, making them powerful models for investigating development, disease mechanisms, pharmacological testing, and host–microbe interactions. Although standardized protocols for chicken intestinal organoids have been established, a defined culture system for [...] Read more.
Intestinal organoids possess self-organizing capacity and recapitulate essential features of intestinal architecture and function, making them powerful models for investigating development, disease mechanisms, pharmacological testing, and host–microbe interactions. Although standardized protocols for chicken intestinal organoids have been established, a defined culture system for ducks has not been available. In this study, we optimized crypt isolation procedures and culture medium composition to establish a reproducible system tailored to duck intestinal stem cells. Among various digestive solutions, ethylene glycol tetraacetic acid (EGTA) achieved the highest crypt isolation efficacy and organoid survival. Suspension culture resulted in better survival, proliferation, and differentiation of intestinal stem cells than air–liquid interface and embedding methods (p < 0.05). Immunofluorescence and real-time PCR indicated the presence of multiple epithelial lineages, including stem cells, Paneth cells, enterocytes, goblet cells, and enteroendocrine cells. Media supplemented with CHIR99021 and LDN193189 (CL) supported growth comparable to that of media with EGF, Noggin, and R-spondin 1 (ENR). Duckling serum and specific factors, such as SB203580 and retinol, further improved organoid formation and promoted differentiation. While long-term passaging and expansion remain technically challenging, this work provides the first duck intestinal organoid model and lays the foundation for future applications in avian intestinal research, including nutrition, disease modeling, and intervention strategies. Full article
(This article belongs to the Section Poultry)
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14 pages, 952 KB  
Article
Association Analysis of the FTO Gene Polymorphisms with Growth and Carcass Traits of Heying Black Chicken and Tissue Expression Profile
by Hao Ding, Lan Chen, Can Chen, Tao Zhang, Weilin Chen, Genxi Zhang, Jinyu Wang and Kaizhou Xie
Animals 2025, 15(18), 2718; https://doi.org/10.3390/ani15182718 - 17 Sep 2025
Viewed by 1002
Abstract
Fat volume and obesity-related genes (e.g., the FTO gene) are important candidate genes affecting energy metabolism. Single nucleotide polymorphisms (SNPs) in the FTO gene are associated with carcass, growth and meat quality traits of pigs, cattle, sheep, rabbits and ducks. The purpose of [...] Read more.
Fat volume and obesity-related genes (e.g., the FTO gene) are important candidate genes affecting energy metabolism. Single nucleotide polymorphisms (SNPs) in the FTO gene are associated with carcass, growth and meat quality traits of pigs, cattle, sheep, rabbits and ducks. The purpose of this study was to detect the single nucleotide polymorphisms in the chicken FTO gene coding region by DNA sequencing and analyze its association with the carcass and growth traits of Heying black chickens. We detected polymorphisms in exons 5, 7, 8 and 9, respectively, g.57337C>A, g.64757T>G, g.97213G>A and g.220985G>A, which are synonymous mutations. g.57337C>A mutation site CA and AA genotype individuals were significantly higher than CC genotype individuals in live weights, head weights, breast muscle weights and leg muscle weights (p < 0.05), AA genotype individuals were significantly higher than CC genotype individuals in slaughter live weights and liver weights (p < 0.05) and CA and AA genotype individuals were significantly higher than CC genotype individuals in heart weights (p < 0.01). In terms of growth traits, the weights of individuals with genotype CA at 8 weeks were significantly higher than that of individuals with genotype CC (p < 0.05), and the weights of individuals with genotype AA and CA at 10 weeks and 16 weeks were significantly higher than that of individuals with genotype CC (p < 0.05). For the g.64757T > G mutation, individuals with the TT genotype exhibited significantly higher values (p < 0.05) than those with the TG genotype across multiple traits, including slaughter weight, live weight, eviscerated weight and semi-eviscerated weight. Individuals with GG genotypes were significantly higher than individuals with TG genotypes (p < 0.05) in slaughter weights and wing weights. In terms of growth traits, the 16-week-old body weight of individuals with TT genotype was significantly higher than that of individuals with TG genotype (p < 0.01). The GA genotype exhibited significantly higher slaughter weights in the g.97213G>A variant compared to the GG genotype (p < 0.01), and in live weights, eviscerated weights, semi-eviscerated weights, leg muscle weights and wing weights, GA genotype was significantly higher than in GG genotype (p < 0.05). In terms of growth traits, GA genotype was significantly higher in individuals 8 weeks old, 10 weeks old and 16 weeks old than GG genotype (p < 0.05). g.220985G>A was significantly higher in individuals with GG genotype than GA genotype (p < 0.05). In terms of growth traits, the weight of GG genotype was significantly higher than that of GA genotype (p < 0.05). The results showed that the FTO gene may be a candidate gene related to chicken growth and slaughter traits and lays a foundation for Heying black chicken assisted breeding. Full article
(This article belongs to the Section Poultry)
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19 pages, 17140 KB  
Article
Chinese Herbal Medicine Compound Microecological Agent (C-MEA) Improves Egg Production Performance in Caged Laying Ducks via Microbiota–Gut–Ovary Axis
by Yanfeng Lu, Lei Zhang, Rui Zhu, Xiujun Duan, Guobo Sun and Yuying Jiang
Vet. Sci. 2025, 12(9), 808; https://doi.org/10.3390/vetsci12090808 - 25 Aug 2025
Viewed by 1740
Abstract
This study was conducted to investigate the effects of a Chinese herbal medicine compound microecological agent (C-MEA) on the egg production performance, ovarian follicle development, ovary transcriptome, and cecal microbiota of caged laying ducks. A total of 108 black Muscovy ducks (150 days [...] Read more.
This study was conducted to investigate the effects of a Chinese herbal medicine compound microecological agent (C-MEA) on the egg production performance, ovarian follicle development, ovary transcriptome, and cecal microbiota of caged laying ducks. A total of 108 black Muscovy ducks (150 days old) were randomly divided into three groups for 30 days in a formal feeding trial. Compared with the control basic diet (Group C) and 16 g/kg C-MEA dosage (Group B), the 8 g/kg C-MEA dosage (Group A) increased egg production (average laying rate 69.35%) and follicle development (5~7 Fs, 6~7 LYFs, 11~13 SYFs) mass (p < 0.05). According to RNA-Seq, the ovaries’ transcriptome among different dietary groups enriched six key pathways, including neuroactive ligand–receptor interaction, the PPAR signaling pathway, ECM–receptor interaction, focal adhesion, the adherens junction, and the FoxO signaling pathway, as well as 46 candidate key genes. According to 16S-Seq, the microbial diversity was significantly increased in Group A, and the genus abundances of Sphaerochaeta and UCG-004 were significantly changed among different dietary groups (p < 0.05). Supplementation with C-MEA may optimize the cecal microflora and the interactions between the intestinal microflora and the host. The results from combining RNA-Seq and 16S-Seq demonstrated that the relationship between Sphaerochaeta and the hub gene cluster (F2, KNG1, C5, PLG, F2RL1, FABP1, and GCG) is the most prominent. In conclusion, the egg performance of caged laying ducks can be modulated through the microbiota–gut–ovary axis. Our findings provide new insights for improving gut health and reproductive performance of caged laying ducks. Full article
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Article
Fermented Mixed Feed Increased Egg Quality and Intestinal Health of Laying Ducks
by Changfeng Xiao, Yunying Xu, Changsuo Yang, Daqian He and Lihui Zhu
Agriculture 2025, 15(11), 1230; https://doi.org/10.3390/agriculture15111230 - 5 Jun 2025
Viewed by 2508
Abstract
This study investigated the effects of adding fermented mixed feed (FMF, composed of several unconventional protein feeds, such as brown rice, rice bran, rice bran meal, sunflower meal, cottonseed meal, and corn starch residue) into the diet of Longyan Shan-ma ducks on their [...] Read more.
This study investigated the effects of adding fermented mixed feed (FMF, composed of several unconventional protein feeds, such as brown rice, rice bran, rice bran meal, sunflower meal, cottonseed meal, and corn starch residue) into the diet of Longyan Shan-ma ducks on their egg quality and intestinal health. The ducks were randomly divided into two groups: one group served as the control and received a standard diet, while the other group received a diet in which 4% of the feed was substituted with FMF. Compared to unfermented feed, FMF had elevated lactic acid levels and reduced phytic acid and crude fiber, along with higher amounts of crude protein and a range of amino acids, including serine, histidine, arginine, alanine, valine, methionine, cysteine, isoleucine, and lysine. FMF significantly enhanced egg production and improved the overall egg quality, such as eggshell strength and thickness. It also enhanced total antioxidant capacity and glutathione peroxidase concentrations in serum while reducing serum urea nitrogen and interleukin-1β levels. Histological analysis showed that FMF supplementation improved the ileal villus height-to-crypt depth ratio. Microbiota analysis demonstrated that FMF had a significant impact on β-diversity by increasing Firmicutes, Actinobacteriota, and Desulfobacterota and decreasing Proteobacteria and Myxococcota at the phylum level. The abundance of Corynebacterium, Lactobacillus, and Gallicola was found to be elevated due to FMF at the genus level, whereas Kocuria, Rothia, Helicobacter, and Escherichia-Shigella were decreased. Additionally, diets supplemented with FMF resulted in higher intestinal valeric acid levels among ducks. Our findings indicate that incorporating FMF into laying duck diets can enhance production performance, egg quality, and gut health. Full article
(This article belongs to the Section Farm Animal Production)
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