Animals

24 pages, 6488 KB

Open AccessArticle

Integrated Single-Cell and Bulk Transcriptomics Unveils Immune Profiles in Chick Erythroid Cells upon Avian Pathogenic Escherichia coli Infection

by Fujuan Cai, Xianjue Wang, Chunzhi Wang, Yuzhen Wang and Wenguang Zhang

Animals 2026, 16(2), 179; https://doi.org/10.3390/ani16020179 (registering DOI) - 7 Jan 2026

Abstract

Nucleated erythroid cells (NECs) have emerged as active participants in immune responses in addition to their canonical oxygen transport function. The subpopulations and immune heterogeneity of chick erythroid cells (ch-ECs) upon infection have not been fully characterized. Single-cell RNA sequencing (scRNA-seq) was used [...] Read more.

Nucleated erythroid cells (NECs) have emerged as active participants in immune responses in addition to their canonical oxygen transport function. The subpopulations and immune heterogeneity of chick erythroid cells (ch-ECs) upon infection have not been fully characterized. Single-cell RNA sequencing (scRNA-seq) was used to profile ch-ECs in chicks infected with avian pathogenic Escherichia coli (APEC). Unsupervised clustering uncovered ten distinct ch-EC subpopulations (C1–C10), with significant compositional shifts between infected and control groups. Pseudotime analysis revealed a developmental continuum: C1, C3, C5, and C9 as early progenitors; C2, C4, C6, C7, and C10 as mature erythroid cells; and C8 as a naive population. We revealed 62 immune-related genes, including protein kinases and heat shock proteins, and subpopulation-specific differentially expressed genes (DEGs) linked to immune functions. SCENIC analysis revealed Fos, Srf, and Stat3 as key transcription factors with elevated regulon activity and specificity following infection. Subpopulations C2, C4, C6, and C7, which exhibited marked abundance changes, were scrutinized for immune relevance through integrated multi-omics analysis. Immune-related genes including FOS, AKAP9, HS6ST1, GAB3, TFRC, HSPA8, HSP90AA1, and DNAJB6 were identified. Enrichment analysis indicated activation of the MHC class I antigen presentation pathway, while pathways such as Mitogen-Activated Protein Kinase (MAPK) signaling, NOD-like receptor (NLR) signaling, and the heat shock response were found to be suppressed. In conclusion, this study delineates the immune gene repertoire and signaling networks of ch-ECs during APEC infection, offering new perspectives on NEC immunoregulatory functions. Full article

(This article belongs to the Special Issue Bacterial Disease Research in Livestock and Poultry)

14 pages, 955 KB

Open AccessArticle

The Effects of Reduced Glutathione on Growth Performance, Intestinal Inflammation, and Gut Microbiota in Immune-Stressed Broiler Chickens

by Xin-Qi Wang, Tao Zhang, Ying-Kun Liu, Hao-Jia Li, Kabelo Anthony Makatjane, Zhen Lai, Jian-Xin Bi, Hai-Zhu Zhou and Wei Guo

Animals 2026, 16(2), 178; https://doi.org/10.3390/ani16020178 (registering DOI) - 7 Jan 2026

Abstract

In intensive farming systems, broilers are exposed to various stressors that trigger immune stress. Reduced glutathione is known to play a crucial role in modulating immune function. This study investigated the effects of different doses of reduced glutathione on the growth performance, intestinal [...] Read more.

In intensive farming systems, broilers are exposed to various stressors that trigger immune stress. Reduced glutathione is known to play a crucial role in modulating immune function. This study investigated the effects of different doses of reduced glutathione on the growth performance, intestinal immune function, and gut microbiota of broilers under immune stress. One-day-old broilers were randomly assigned to five groups: group K (control), group L (lipopolysaccharide), and three glutathione supplementation groups, Y1 (50 mg/kg), Y2 (100 mg/kg), and Y3 (200 mg/kg). Each group had four replicates, with 11 broilers per replicate. On days 16, 18, and 20, broilers in groups L and Y1-Y3 received intraperitoneal injections of 0.5 mg/kg of lipopolysaccharide, while group K received an equal volume of saline. On day 16, the average daily weight gain and final body weight of group Y3 were significantly higher than those of groups K and L, whereas on day 21, the average daily weight gain of group Y3 was still significantly higher than that of group L. On day 21 broilers were slaughtered, and samples were collected. In the jejunal mucosa, group Y3 showed significantly reduced levels of IL-2, IL-4, IL-1β, and TNF-α compared to group L. Additionally, group Y3 exhibited reduced relative expression levels of NF-κB, TLR4, IFN-γ, IL-1β, IL-6, IL-4, TNF-α, and IL-2 mRNA; a decreased abundance of Enterococcus, Proteobacteria, and Actinobacteria; and a restored abundance of Ligilactobacillus, Turicibacter, and Firmicutes to normal levels. These findings suggest that 200 mg/kg of reduced glutathione is the optimal dose for improving the gut microbiota composition and mitigating gut damage caused by immune stress by inhibiting the TLR4/NF-κB signaling pathway. Full article

(This article belongs to the Special Issue Feed Additives in Animal Nutrition)

21 pages, 1866 KB

Open AccessArticle

Lean DLY Pig-Derived Fecal Microbiota Promotes Growth Performance by Modulating Gut Microbiota: Serum Metabolic Profiles in Obese Ningxiang Pigs

by Li Han, Feng Zhou, Chen Zhang, Hongkun Li, Yongmin Zheng, Yv Tian, Yang Liu, Jie Yin and Xingguo Huang

Animals 2026, 16(2), 177; https://doi.org/10.3390/ani16020177 (registering DOI) - 7 Jan 2026

Abstract

Fecal microbiota transplantation (FMT) has demonstrated potential in reshaping gut microbiota to improve animal phenotypes, yet its application in lean-type to obese-type pigs like Ningxiang (NX) pigs remains unclear. To address this, we investigated the effects of Lean Duroc × Landrace × Yorkshire [...] Read more.

Fecal microbiota transplantation (FMT) has demonstrated potential in reshaping gut microbiota to improve animal phenotypes, yet its application in lean-type to obese-type pigs like Ningxiang (NX) pigs remains unclear. To address this, we investigated the effects of Lean Duroc × Landrace × Yorkshire (DLY) pig-derived fecal microbiota on the growth, gut microbiota composition, and serum metabolism of obese NX pigs. Thirty-six 50-day-old castrated male NX pigs of similar initial body weight were randomly assigned to either a control group or FMT group. The trial lasted for 35 days. Results indicated that FMT significantly improved the average daily gain and increased nutrient digestibility. Serum biochemical analysis revealed elevated levels of globulin and total protein and reduced low-density lipoprotein cholesterol in the FMT group. In addition, 16S rRNA sequencing demonstrated that FMT modified gut microbiota composition and diversity, enriching beneficial genera such as Blautia, Agathobacter, Faecalibacterium, and Eubacterium_coprostanoligenes_group. Untargeted serum metabolomics further revealed altered metabolite profiles linked to lipid and amino acid metabolism. Correlation analysis further revealed a link between these enriched bacteria and metabolites changes. Overall, these findings demonstrate that transplantation of the fecal microbiota from lean DLY pigs significantly improved the growth performance of obese NX pigs by improving nutrient digestibility and modulating the gut microbiota–host metabolic axis. Full article

(This article belongs to the Section Pigs)

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20 pages, 30451 KB

Open AccessArticle

A Comprehensive Atlas of Testicular lncRNAs Reveals Dynamic Changes and Regulatory Networks During Sexual Maturation in Tibetan Sheep

by Taotao Li, Huihui Wang, Ruirui Luo, Juanjuan Song, Yi Wu, Meng Jia, Yong Zhang and Youji Ma

Animals 2026, 16(2), 176; https://doi.org/10.3390/ani16020176 - 7 Jan 2026

Abstract

Tibetan sheep, a dominant livestock species on the Qinghai–Tibet Plateau, is characterized by late sexual maturity and low reproductive efficiency. Although long non-coding RNAs (lncRNAs) are known to play critical regulatory roles in mammalian testicular development and spermatogenesis, their expression dynamics and functions [...] Read more.

Tibetan sheep, a dominant livestock species on the Qinghai–Tibet Plateau, is characterized by late sexual maturity and low reproductive efficiency. Although long non-coding RNAs (lncRNAs) are known to play critical regulatory roles in mammalian testicular development and spermatogenesis, their expression dynamics and functions in Tibetan sheep remain poorly understood. In this study, we integrated histological and transcriptomic analyses to profile testicular lncRNAs across three developmental stages: pre-pubertal (3 months), sexually mature (1 year), and adult (3 years). Histological examination showed progressive structural maturation of seminiferous tubules, accompanied by significant increases in testicular weight and serum testosterone levels. RNA sequencing identified 10,857 high-confidence lncRNAs and uncovered extensive reprogramming of the lncRNA transcriptome during sexual maturation, with 7784 lncRNAs differentially expressed between pre-pubertal and post-pubertal stages. Functional enrichment analyses of cis- and antisense-target genes indicated that these lncRNAs were involved in key biological processes, including cell cycle regulation, TGF-β and Hippo signaling pathways, extracellular matrix organization, glycolysis, and apoptosis. Co-expression network analysis further linked upregulated lncRNAs to spermatogenesis-related genes involved in processes such as sperm nuclear condensation (e.g., TNP1) and metabolic support (e.g., PFKP). Our findings demonstrated that lncRNAs coordinate testicular development and spermatogenesis in Tibetan sheep by modulating transcriptional networks, remodeling the cellular microenvironment, and reprogramming energy metabolism. This study provides the first comprehensive atlas of testicular lncRNAs in Tibetan sheep and offers novel insights into the epigenetic regulation of male reproduction in high-altitude mammals. Full article

(This article belongs to the Special Issue Male Germ Cell Development in Animals)

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14 pages, 587 KB

Open AccessFeature PaperArticle

Bioacoustic Detection of Wolves Using AI (BirdNET, Cry-Wolf and BioLingual)

by Johanne Holm Jacobsen, Pietro Orlando, Line Østergaard Jensen, Sussie Pagh and Cino Pertoldi

Animals 2026, 16(2), 175; https://doi.org/10.3390/ani16020175 - 7 Jan 2026

Abstract

Rising numbers of wolf (Canis lupus) populations make traditional, resource-intensive methods of wolf monitoring increasingly challenging and often insufficient. This study explores how wolf howls can be used as a new monitoring tool for wolves by applying Artificial Intelligence (AI) methods [...] Read more.

Rising numbers of wolf (Canis lupus) populations make traditional, resource-intensive methods of wolf monitoring increasingly challenging and often insufficient. This study explores how wolf howls can be used as a new monitoring tool for wolves by applying Artificial Intelligence (AI) methods to detect and classify wolf howls from acoustic recordings, thereby improving the effectiveness of wolf population monitoring. Three AI approaches are evaluated: BirdNET, Yellowstone’s Cry-Wolf project system, and BioLingual. Data were collected using Song Meter SM4 (SM4) audio recorders in a known wolf territory in Klelund Dyrehave, Denmark, and manually validated to establish a ground truth of 260 wolf howls. Results demonstrate that while AI solutions currently do not achieve the complete precision or overall accuracy of expert manual analysis, they offer tremendous efficiency gains, significantly reducing processing time. BirdNET achieved the highest recall at 78.5% (204/260 howls detected), though with a low precision of 0.007 (resulting in 28,773 false positives). BioLingual detected 61.5% of howls (160/260) with 0.005 precision (30,163 false positives), and Cry-Wolf detected 59.6% of howls (155/260) with 0.005 precision (30,099 false positives). Crucially, a combined approach utilizing all three models achieved a 96.2% recall (250/260 howls detected). This suggests that while AI solutions primarily function as powerful human-aided data reduction tools rather than fully autonomous detectors, they represent a valuable, scalable, and non-invasive complement to traditional methods in wolf research and conservation, making large-scale monitoring more feasible. Full article

(This article belongs to the Section Animal System and Management)

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16 pages, 312 KB

Open AccessFeature PaperArticle

Curcumin as a Dietary Additive in Early-Finished Feedlot Steers and Its Effects on Performance, Ruminal Environment, Animal Health, and Meat Quality

by Maisa Damo, João Gustavo Weschenfelder Wandscheer, Mateus Henrique Signor, Charles Marcon, Luisa Nora, Ana Carolina Hadlich Xavier, Roger Wagner, Marcelo Vedovatto and Aleksandro Schafer da Silva

Animals 2026, 16(2), 174; https://doi.org/10.3390/ani16020174 - 7 Jan 2026

Abstract

The present study aimed to evaluate whether the inclusion of curcumin as a performance enhancer in finishing cattle has positive effects on animal production, the ruminal environment, hematological and biochemical markers, as well as meat quality. Sixteen castrated Holstein steers, 8 months of [...] Read more.

The present study aimed to evaluate whether the inclusion of curcumin as a performance enhancer in finishing cattle has positive effects on animal production, the ruminal environment, hematological and biochemical markers, as well as meat quality. Sixteen castrated Holstein steers, 8 months of age, with an average body weight of 247 ± 3.89 kg were divided into two groups: Control, consisting of animals that consumed 144 mg of monensin/animal/day (n = 8); Treatment, consisting of animals that consumed 552 mg of curcumin/animal/day (n = 8). Animals were fed a diet containing 44% roughage (corn silage) and 56% concentrate for 105 days. During this period, body weight measurements were recorded, and blood samples were collected for the determination of hematological, biochemical, and immunological variables. At the end of the experimental period, animals were slaughtered and meat samples were collected for evaluation. No differences were observed between groups for animal performance, feed intake, or feed efficiency. A lower leukocyte count (lymphocyte and granulocyte) and a higher platelet count were observed in animals that consumed curcumin. These animals also exhibited higher cholesterol levels, along with lower circulating glucose concentrations compared to the control group. In the ruminal environment, higher bacterial activity and greater protozoal counts were observed in the treatment group, but no effects on ruminal short-chain fatty acids were observed. Higher activity of the enzymes glutathione S-transferase in serum and superoxide dismutase in meat was observed, combined with lower lipid peroxidation in serum and meat. Meat from steers fed curcumin showed greater yellow color intensity, higher water-holding capacity, and a higher proportion of monounsaturated fatty acids compared to the control group. These results suggest that curcumin can be used as a performance enhancer, similar to monensin, when thinking about performance, but beyond that, curcumin triggered anti-inflammatory and antioxidant action. Full article

(This article belongs to the Special Issue Feed Additives in Animal Nutrition)

15 pages, 761 KB

Open AccessArticle

Protective Effects of Humic Acid on Intestinal Barrier Dysfunction and Inflammatory Activation in Canine Cell-Based Models

by Alma Virág Móritz, Orsolya Farkas, Ákos Jerzsele and Nikolett Palkovicsné Pézsa

Animals 2026, 16(2), 173; https://doi.org/10.3390/ani16020173 - 7 Jan 2026

Abstract

The intestinal barrier is essential for gastrointestinal and systemic homeostasis by enabling nutrient absorption while limiting the translocation of pathogens and toxins. When barrier function is impaired, bacterial components such as lipopolysaccharides (LPSs) may cross the epithelium and promote inflammatory signaling. In dogs, [...] Read more.

The intestinal barrier is essential for gastrointestinal and systemic homeostasis by enabling nutrient absorption while limiting the translocation of pathogens and toxins. When barrier function is impaired, bacterial components such as lipopolysaccharides (LPSs) may cross the epithelium and promote inflammatory signaling. In dogs, chronic inflammatory enteropathies are frequent disorders associated with barrier dysfunction, dysbiosis, and immune dysregulation, and may progress to protein-losing enteropathy or systemic inflammation. Humic substances, particularly humic acid (HA), are natural organic compounds with reported antioxidative, immunomodulatory, and barrier-supporting effects; however, the cellular mechanisms underlying these effects in intestinal and immune models remain insufficiently characterized. This study evaluated the effects of a commercially available HA-based supplement on epithelial barrier integrity and inflammatory responses using an in vitro system combining IPEC-J2 intestinal epithelial cells and primary canine peripheral blood mononuclear cells (PBMCs). Epithelial barrier integrity (FD4 paracellular flux), reactive oxygen species, and cytokine production (TNF-α, IL-6) were assessed under basal and LPS-stimulated conditions. HA treatment preserved epithelial barrier function and reduced LPS-induced pro-inflammatory cytokine production, supporting further investigation of HA as a nutraceutical adjunct for gut health support in dogs with chronic enteropathies. Full article

(This article belongs to the Section Companion Animals)

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22 pages, 6894 KB

Open AccessArticle

Genome-Wide Characterization of Four Gastropod Species Ionotropic Receptors Reveals Diet-Linked Evolutionary Patterns of Functional Divergence

by Gang Wang, Yi-Qi Sun, Fang Wang, Zhi-Yong Wang, Ni-Ying Sun, Meng-Jun Wei, Yu-Tong Shen, Yi-Jia Li, Quan-Qing Sun, Yushinta Fujaya, Xun-Guang Bian, Wen-Qi Yang and Kianann Tan

Animals 2026, 16(2), 172; https://doi.org/10.3390/ani16020172 - 7 Jan 2026

Abstract

Ionotropic receptors (IRs) are a divergent subfamily of ionotropic glutamate receptors (iGluRs) that detect olfactory and environmental cues, influencing behaviors such as foraging and adaptation. To explore the evolution of IRs in relation to feeding ecology, we identified IRs and iGluRs from the [...] Read more.

Ionotropic receptors (IRs) are a divergent subfamily of ionotropic glutamate receptors (iGluRs) that detect olfactory and environmental cues, influencing behaviors such as foraging and adaptation. To explore the evolution of IRs in relation to feeding ecology, we identified IRs and iGluRs from the genomes of four gastropods with distinct diets: Pomacea canaliculata (9 IRs/18 iGluRs), Bellamya purificata (10/22), Cipangopaludina chinensis (11/23), and Babylonia areolata (22/41). IRs were markedly expanded in B. areolata, suggesting lineage-specific diversification. Phylogenetic analysis grouped IRs and iGluRs into three clades, with IRs clustered with GluD, supporting early functional divergence following gene duplication. In all species, IR25b showed tandem duplication and played a central role in protein–protein interaction (PPI) networks. Most IRs were acidic, whereas IR-A and IR-C subgroups were basic, suggesting functional specialization among subfamilies. Structural analysis showed that IRs share conserved domains and motifs across species. Most IRs experienced purifying selection, while P. canaliculata showed relaxed constraints, suggesting weaker functional limitation. Collinearity analysis identified conserved genes, such as BarIR-A.6 and BarIR-D.1, across species. qPCR confirmed tissue-specific expression of IRs in multiple organs. Together, these results reveal the molecular features and evolutionary patterns of IRs in gastropods, highlighting their potential roles in olfaction and dietary adaptation. Full article

(This article belongs to the Section Animal Genetics and Genomics)

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12 pages, 10620 KB

Open AccessCase Report

Tracheal External Support Using a Polypropylene Linear Prosthesis for Tracheal Stent Fracture in a Dog: A Case Report

by Tomohiro Yoshida, Ryou Tanaka, Kazuyuki Terai, Aki Takeuchi, Akari Hatanaka, Daisuke Ito and Takashi Tanaka

Animals 2026, 16(2), 171; https://doi.org/10.3390/ani16020171 - 7 Jan 2026

Abstract

Tracheal stent fracture is a major complication of endoluminal tracheal stent (ELS) for canine tracheal collapse, and optimal management strategies remain unclear. A 4-year-old Yorkshire Terrier presented with respiratory distress caused by complete ELS fracture. Imaging and bronchoscopy revealed intraluminal protrusion of fractured [...] Read more.

Tracheal stent fracture is a major complication of endoluminal tracheal stent (ELS) for canine tracheal collapse, and optimal management strategies remain unclear. A 4-year-old Yorkshire Terrier presented with respiratory distress caused by complete ELS fracture. Imaging and bronchoscopy revealed intraluminal protrusion of fractured stent segments, ventral tracheal cartilage invagination, and marked luminal deformation. A polypropylene linear prosthesis (PLLP) was selected as an external tracheal support. Its continuous band-like structure allowed broad and uniform reinforcement of the tracheal wall and redistribution of mechanical stress. PLLP placement successfully restored a near-normal tracheal contour and stabilized the fractured stent without introducing additional intraluminal material. Postoperative bronchoscopy confirmed improved tracheal patency, and no further deformation or stent damage was observed despite several months of altered airway dynamics associated with laryngeal paralysis. This case suggests that PLLP may represent a valid surgical option for managing tracheal stent fracture in dogs. Full article

(This article belongs to the Special Issue Surgical Procedures and Postoperative Complications in Animals)

19 pages, 9572 KB

Open AccessArticle

Development of a Liquid Microbial Enzyme Synergistic Fermentation Process for Strong-Aroma and Soy-Sauce-Aroma Fresh Distiller’s Grains and the Evaluation of Their Nutritional Value in Pigs

by Caimei Wu, Ziyun Zhou, Meihong Li, Kun Zhang, Yixuan Zhou, Fali Wu, Jie Yu, Jian Li, Ruinan Zhang, Hua Li, Jiayong Tang, David Thomas, Lianqiang Che and Yang Lyu

Animals 2026, 16(2), 170; https://doi.org/10.3390/ani16020170 - 7 Jan 2026

Abstract

China annually produces around 20 million tons of distiller’s grains, whose utilization is important for resource efficiency. These grains are rich in crude protein (CP), crude fiber (CF), and other nutrients, though their composition varies by grain type. This study applied a microbial–enzyme [...] Read more.

China annually produces around 20 million tons of distiller’s grains, whose utilization is important for resource efficiency. These grains are rich in crude protein (CP), crude fiber (CF), and other nutrients, though their composition varies by grain type. This study applied a microbial–enzyme synergistic liquid fermentation process to two types—strong-aroma and soy-sauce-aroma grains—using a composite inoculant (Lactobacillus, Saccharomyces cerevisiae, Bacillus, each >1 × 10⁹ CFU/g) along with xylanase, cellulase, and protease. Optimal fermentation conditions were identified: for strong-aroma grains—water-to-grain material ratio 1.8, inoculum 0.25%, 25 °C, 16 h; for soy-sauce-aroma grains—ratio 1.8, inoculum 0.3%, 25.5 °C, 13 h. After fermentation, CP increased by 13.62% and CF decreased by 30.37% in strong-aroma grains, while in soy-sauce-aroma grains an 8.83% CP increase and 31.31% CF reduction were observed. Structural analysis of both grains showed looser fibrous formations and changes in protein secondary structure. Full article

(This article belongs to the Special Issue Use of Agro-Industrial Co-Products in Animal Nutrition)

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16 pages, 2008 KB

Open AccessArticle

Genomic Differentiation and Diversity in Persian Gulf Hawksbill Turtles (Eretmochelys imbricata) Revealed by the First Whole-Genome Sequencing Study

by Mohammadali Farahvashi, Mohammadreza Mohammadabadi, Majid Askari-Hesni, Zeinab Amiri Ghanatsaman and Hojjat Asadollahpour Nanaei

Animals 2026, 16(2), 169; https://doi.org/10.3390/ani16020169 - 7 Jan 2026

Abstract

This is the first WGS study of the hawksbill sea turtle (Eretmochelys imbricata) from the Persian Gulf. Sequencing of 17 individuals from four nesting islands in the southern part of Iran’s coastline revealed population-level genetic differentiation with consistently low rates of [...] Read more.

This is the first WGS study of the hawksbill sea turtle (Eretmochelys imbricata) from the Persian Gulf. Sequencing of 17 individuals from four nesting islands in the southern part of Iran’s coastline revealed population-level genetic differentiation with consistently low rates of nucleic diversity and differentiated demographic footprints in sampling populations in the Persian Gulf. In a population structure analysis, four populations were discovered; Shidvar had long-term isolation and Nakhiloo demonstrated admixture and higher heterogeneity. ROH and LD profiles are consistent with past bottlenecks, rather than recent inbreeding, and reflect each island’s demographic history. The observed fine-scale genomic divergence (e.g., even between sites in close proximity, such as Ommolgorm and Nakhiloo) suggests that natal homing and local environmental selection are meaningful drivers of genetic differentiation, and that specific conservation strategies should be implemented at each site. Full article

(This article belongs to the Section Animal Genetics and Genomics)

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13 pages, 739 KB

Open AccessArticle

Factors Influencing the Production Efficiency of Cloned Pigs: A Large-Scale Retrospective Analysis

by Huaxing Zhao, Shouquan Zhang, Xiaopeng Tang, Rong Zhou, Ranbiao Mai, Lvhua Luo, Qiaoyun Su, Sixiu Huang, Zhenfang Wu, Zicong Li, Gengyuan Cai and Junsong Shi

Animals 2026, 16(2), 168; https://doi.org/10.3390/ani16020168 - 7 Jan 2026

Abstract

Pig somatic cell nuclear transfer (SCNT) has valuable applications in agriculture, biomedicine, and life sciences, yet low cloning efficiency remains a major constraint limiting its application. To systematically investigate factors related to the production efficiency of pig cloning, this study conducted a retrospective [...] Read more.

Pig somatic cell nuclear transfer (SCNT) has valuable applications in agriculture, biomedicine, and life sciences, yet low cloning efficiency remains a major constraint limiting its application. To systematically investigate factors related to the production efficiency of pig cloning, this study conducted a retrospective analysis of 367,701 SCNT embryos transferred into 2019 surrogate sows over five years, focusing on breeds of donor cells, the season of embryo transfers, and the number of embryos transferred per surrogate. Our data demonstrate that the genetic background of donor cells is a critical determinant. SCNT embryos generated by wild-type (WT) Pietrain and Duroc pigs yielded significantly higher cloning efficiencies compared to those from Large White and Yorkshire pigs. This breed-specific influence was also observed with genetically modified (GM) donor cells. Nevertheless, within the GM groups, GM-Duroc and GM-Yorkshire showed superior efficiency compared to GM-Large White and GM-Bama. Furthermore, Summer was identified as the least favorable season for embryo transfer, with significantly lower pregnancy rates, delivery rates, and cloning efficiency compared to the other seasons. Importantly, we established that transferring 100–150 embryos per recipient optimized cloning efficiency, significantly outperforming groups receiving higher embryo numbers without compromising pregnancy rates, delivery rates, or average litter sizes. Our findings provide valuable guidance for optimizing large-scale SCNT protocols in swine. Full article

(This article belongs to the Special Issue Assisted Reproductive Technologies in Production Animals)

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17 pages, 860 KB

Open AccessArticle

Effects of Fermentation Compound Chinese Herbal Medicine on the Reproductive Performance, Immune and Antioxidant Status, and Colostrum Metabolites of Ningxiang Sows During the Lactation Period

by Qingtai Zhang, Haibo Huang, Xinhao Song, Weiguang Yang, Rejun Fang and Chengkun Fang

Animals 2026, 16(2), 167; https://doi.org/10.3390/ani16020167 - 7 Jan 2026

Abstract

The present study aimed to evaluate the effects of fermented compound Chinese herbal medicine (FCHM, Vaccaria segetali, Tetrapanax papyriferus, Ligusticum chuanxiong Hort and Rhaponticum uniflorum) on the reproductive performance of Ningxiang sows. A total of 30 Ningxiang sows were randomly [...] Read more.

The present study aimed to evaluate the effects of fermented compound Chinese herbal medicine (FCHM, Vaccaria segetali, Tetrapanax papyriferus, Ligusticum chuanxiong Hort and Rhaponticum uniflorum) on the reproductive performance of Ningxiang sows. A total of 30 Ningxiang sows were randomly assigned to the control group (CON), CHM group, and FCHM group. The results indicated that dietary CHMs and FCHM supplementation significantly increased (p < 0.05) the milk production, feed intake, reproductive performance protein content, and lactose content in the milk of sows and significantly decreased (p < 0.05) the number of somatic cells in colostrum and in the number of low-birthweight piglets. Dietary FCHM supplementation significantly increased (p < 0.05) the number of healthy piglets, birth litter weight, colostrum fat content, and feed intake. Moreover, malondialdehyde, porcine interleukin-1α, and porcine interleukin-6 in sow serum were decreased (p < 0.05), and the activity of superoxide dismutase, total antioxidant capacity, immunoglobulin, prolactin and progesterone levels (p < 0.05) were increased, with FCHM supplementation. The colostrum metabolomics analyses showed that FCHM significantly enriched the oxytocin signaling pathway, calcium signaling pathway, and pathways associated with milk composition synthesis. In conclusion, supplementing with FCHM improved the reproductive performance and milk metabolic biomarkers of sows and may serve as an effective feed additive to improve productivity. Full article

(This article belongs to the Special Issue Feed Additives and Micronutrients on Performance, Nutrient Utilization, Immunity, Oxidative Stress, Gut Health and Microbiome of Pigs and Sows: Third Edition)

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13 pages, 7051 KB

Open AccessArticle

Morphological, Molecular and Phylogenetic Characterization of Ceratomyxa nemiptera sp. nov. (Myxozoa: Ceratomyxidae) Infecting Nemipterus virgatus Houttuyn, 1782 in the East China Sea

by Pingping Li, Yang Zhou, Xiaoping Tan, Yuanjun Zhao and Chengzhong Yang

Animals 2026, 16(2), 166; https://doi.org/10.3390/ani16020166 - 7 Jan 2026

Abstract

A newly discovered myxosporean parasite was described from the gallbladder of Nemipterus virgatus Houttuyn, 1782 collected from the East China Sea. Mature myxospores are crescent-shaped with shell valves that taper gradually toward rounded ends. Each myxospore contained two sub-spherical polar capsules located near [...] Read more.

A newly discovered myxosporean parasite was described from the gallbladder of Nemipterus virgatus Houttuyn, 1782 collected from the East China Sea. Mature myxospores are crescent-shaped with shell valves that taper gradually toward rounded ends. Each myxospore contained two sub-spherical polar capsules located near the anterior end, closely aligned along the suture line. The mature myxospores measured 6.2 ± 0.6 (5.4–6.9) μm in length and 44.8 ± 4.6 (38.5–53.1) μm in thickness. Polar capsules measured 2.8 ± 0.2 (2.4–3.1) μm in length and 2.3 ± 0.2 (1.9–2.6) μm in width, with polar filaments coiled in 2–3 turns. The small subunit ribosomal DNA (SSU rDNA) sequence of Ceratomyxa nemiptera sp. nov was distinct from all known myxosporeans, showing the highest similarity (93.56%) and the shortest genetic distance (0.0637) with Ceratomyxa arcuata Thélohan, 1892. The phylogenetic analysis revealed that C. nemiptera sp. nov. was positioned within a later-diverging lineage, forming a sister-group relationship with a clade containing C. arcuata and Ceratomyxa cretensis Kalatzis, Kokkari & Katharios, 2013. This is the first report of a Ceratomyxa species infecting N. virgatus. Full article

(This article belongs to the Section Aquatic Animals)

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52 pages, 484 KB

Open AccessReview

Scoping Review of the Socioeconomic Value of Working Equids, and the Impact of Educational Interventions Aimed at Improving Their Welfare

by Amelia Cameron, Sarah L. Freeman, Isabella Wild, Jessica Burridge and Katie Burrell

Animals 2026, 16(2), 165; https://doi.org/10.3390/ani16020165 - 7 Jan 2026

Abstract

Working equids support millions of people globally, especially in low-income, lower-middle-income, and upper-middle-income countries. However, they commonly suffer from poor welfare and are typically overlooked in policy and funding decisions. This scoping review aimed to collate evidence on two topics related to working [...] Read more.

Working equids support millions of people globally, especially in low-income, lower-middle-income, and upper-middle-income countries. However, they commonly suffer from poor welfare and are typically overlooked in policy and funding decisions. This scoping review aimed to collate evidence on two topics related to working equid use in low- and middle-income countries: their socioeconomic value to their owners and the impact of educational interventions for owners/handlers aiming to improve equid welfare. Original research published from 2014 onwards was eligible for inclusion. This scoping review followed the JBI methodology and PRISMA-ScR framework. One search strategy encompassing both topics was applied to five databases (CAB Abstracts, MEDLINE, Embase, Web of Science, and IBSS) on 24.04.24. Key characteristics and findings of eligible studies were charted. In total, 3514 sources were independently screened by two reviewers. In total, 61 socioeconomic value studies (47 journal articles, 2 reports, and 12 conference contributions) and 23 educational intervention studies (11 journal articles and 12 conference contributions) were included. Working equids supported their owners’ livelihoods in wide-ranging ways and contributed to the United Nations’ Sustainable Development Goals. Educational interventions employed varied approaches, and most reported success. Multilevel initiatives and those developed through participatory engagement may be more likely to directly improve equid welfare in the long term. These aspects should be prioritised during intervention development. The included studies used inconsistent terminology and were of variable quality. This review highlights the importance of including working equids within policy and funding strategies and provides recommendations to increase the discoverability, quality, and impact of working equid research. Full article

(This article belongs to the Special Issue Working Equids: Welfare, Health and Behavior)

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15 pages, 2810 KB

Open AccessArticle

Investigation of Mechanism of Small Peptide Application in Enhancing Laying Performance of Late-Laying Hens Through Bidirectional Liver–Gut Interactions

by Yuanyuan Li, Xiaopeng Liao, Xiaoyue Wang, Yiping Wang, Qin Liu, Lizhi Li, Dongsheng Guo and Zhen Li

Animals 2026, 16(2), 164; https://doi.org/10.3390/ani16020164 - 6 Jan 2026

Abstract

The primary objective of this study was to investigate the mechanism through which small peptides regulate the productive performance and egg quality of laying hens during the late-laying period. A total of 200 Lohmann Pink laying hens, aged 400 days, were randomly assigned [...] Read more.

The primary objective of this study was to investigate the mechanism through which small peptides regulate the productive performance and egg quality of laying hens during the late-laying period. A total of 200 Lohmann Pink laying hens, aged 400 days, were randomly assigned into a control treatment (CON) and a small peptide treatment (SP) for a 120-day treating period. Productive performance, egg quality, serum antioxidant capacity, intestinal morphology, microbial community, and hepatic gene expressions were measured. Results showed that SP supplementation significantly increased eggshell strength and albumen height, while reducing the rate of abnormal eggs (p < 0.05). SP notably enhanced the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and hepatic lipase (p < 0.05). Additionally, SP supplementation significantly increased microbial α-diversity (p < 0.05) and elevated the relative abundances of Ruminococcus, Lactobacillus, and Faecalibacterium (p < 0.05). Hepatic transcriptomic analysis revealed that up-regulated genes in the SP treatment were primarily enriched in steroid biosynthesis, while down-regulated genes were mainly associated with the Yersinia infection pathway. In conclusion, small peptide supplementation efficiently improved eggshell strength and albumen height while reducing the rate of abnormal eggs by modulating the interactions between gut microbiota and hepatic gene expressions. Our findings may provide an effective option for enhancing egg quality in the late-laying period. Full article

(This article belongs to the Special Issue A Paradigm Shift—Advances in Poultry Nutrition and Management, and Alternative Poultry Feed Ingredients Using Novel Feed Processing Methods: Second Edition)

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21 pages, 3397 KB

Open AccessArticle

Aster pekinensis Extract Mitigates High-Fat-Diet-Induced Obesity and Metabolic Dysfunction in Mice

by Hyeon Jeong Moon, Seon-Jin Lee, Geon Woo Kim, Yeong-Bin Baek and Sang-Ik Park

Animals 2026, 16(2), 163; https://doi.org/10.3390/ani16020163 - 6 Jan 2026

Abstract

Obesity and metabolic disorders are an increasing concern in companion animals, creating demand for herb-derived nutraceuticals and functional feeds. This study evaluated whether a water extract of Aster pekinensis (AP) ameliorates high-fat-diet (HFD)-induced obesity and metabolic dysfunction in mice. The phytochemical profile of [...] Read more.

Obesity and metabolic disorders are an increasing concern in companion animals, creating demand for herb-derived nutraceuticals and functional feeds. This study evaluated whether a water extract of Aster pekinensis (AP) ameliorates high-fat-diet (HFD)-induced obesity and metabolic dysfunction in mice. The phytochemical profile of AP was characterized by mass spectrometry, revealing oleanane-type triterpenoid saponins and dicaffeoylquinic acids. Male C57BL/6 mice were fed an HFD and orally given AP (10–200 mg/kg/day) for 12 weeks, with normal diet and untreated HFD groups as controls. AP at 50–200 mg/kg/day reduced body-weight gain, adipose tissue mass and food efficiency without lowering food intake, and improved fasting glucose and atherogenic lipid indices. AP also enhanced glucose tolerance and insulin sensitivity, attenuated hepatic steatosis, hepatocellular ballooning, lobular inflammation and non-alcoholic fatty liver disease (NAFLD) Activity Score, and decreased serum liver enzyme activities. These effects were accompanied by modulation of hepatic genes involved in lipogenesis and inflammation. Together, these findings indicate that AP extract mitigates diet-induced obesity and NAFLD-like liver injury and supports further development as a herb-derived nutraceutical or functional feed ingredient for managing obesity-related metabolic disorders in companion animals. Full article

(This article belongs to the Special Issue Herbal Extracts and Herbal-Derived Products to Support Animal Health and Productivity)

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18 pages, 313 KB

Open AccessArticle

Feeding Low- and High-Fibre Sunflower Meal to Broiler Chickens—Effects of Inclusion Rate and Age of Birds on the Production Traits, Carcass Composition, Nutrient Digestibility, Gut Viscosity, and Caecal Short-Chain Fatty Acid Content

by Kesete Goitom Tewelde, Brigitta Kiss, Tivadar Csiszér, László Pál, Nikoletta Such, Ádám Bartos and Károly Dublecz

Animals 2026, 16(2), 162; https://doi.org/10.3390/ani16020162 - 6 Jan 2026

Abstract

Sunflower meal (SM) is an economically important, inexpensive, and locally abundant alternative protein source in the Eurozone. The study aimed at investigating the effects of feeding two forms of SM on the production traits, carcass composition, nutrient digestibility, and some gut parameters of [...] Read more.

Sunflower meal (SM) is an economically important, inexpensive, and locally abundant alternative protein source in the Eurozone. The study aimed at investigating the effects of feeding two forms of SM on the production traits, carcass composition, nutrient digestibility, and some gut parameters of broiler chickens. A total of 600-day-old Ross 308 male broilers were fed five isocaloric and isonitrogenous diets. Besides the control diet (C), the high- and low-fibre SMs (HFSM and LFSM) were fed at 20% (HFSM20 and LFSM20) and 30% (HFSM30 and LFSM30). Compared to the C, feeding the SM-containing diets did not affect the feed intake (FI) of birds. In the finisher phase, the HFSM30 treatment resulted in significantly higher, while the LFSM30 diet in significantly lower body weight gain. All SM treatments impaired FCR, but the difference was significant only in the grower phase. In comparison with the C, the SM treatments failed to modify carcass composition. The fat digestibility and the AMEn content increased, while the starch digestibility decreased when SM was fed. Except the LFSM30 treatment in the grower phase, the Nitrogen retention of birds was not affected. The SM-containing diets reduced the urinary N excretion, and the total N excretion of growers. Furthermore, the HFSM30 reduced the jejunal viscosity during the grower and finisher phases. The treatments did not modify the short-chain fatty acid contents of the caeca. In conclusion, SM can be used even at 20 and 30% in the nutrition of broiler chickens. However, the responses are affected by the age and the fibre content of SM. Full article

(This article belongs to the Special Issue Optimizing Alternative Protein Sources for Sustainable Poultry Diet)

19 pages, 1499 KB

Open AccessArticle

A Supervised Deep Learning Model Was Developed to Classify Nelore Cattle (Bos indicus) with Heat Stress in the Brazilian Amazon

by Welligton Conceição da Silva, Jamile Andréa Rodrigues da Silva, Lucietta Guerreiro Martorano, Éder Bruno Rebelo da Silva, Cláudio Vieira de Araújo, Raimundo Nonato Colares Camargo-Júnior, Kedson Alessandri Lobo Neves, Tatiane Silva Belo, Leonel António Joaquim, Thomaz Cyro Guimarães de Carvalho Rodrigues, André Guimarães Maciel e Silva and José de Brito Lourenço-Júnior

Animals 2026, 16(2), 161; https://doi.org/10.3390/ani16020161 - 6 Jan 2026

Abstract

Non-invasive and intelligent technologies have been utilized to monitor agricultural systems in real time, facilitating expedient decision-making and the reduction in animal stress in diverse climatic conditions. The objective of this study was to develop a deep learning supervised model to classify Nelore [...] Read more.

Non-invasive and intelligent technologies have been utilized to monitor agricultural systems in real time, facilitating expedient decision-making and the reduction in animal stress in diverse climatic conditions. The objective of this study was to develop a deep learning supervised model to classify Nelore cattle (Bos indicus) into two groups: those in comfort and those under thermal stress. Thirty cattle, aged between 18 and 20 months, were evaluated between June and December 2023, resulting in 676 samples collected across four daily periods (6:00, 12:00, 18:00, and 24:00). Biotic variables included rectal temperature (RT) and respiratory rate (RR), while abiotic variables included air temperature (AT) and relative humidity (RH). The neural network model exhibited an accuracy and recall of 72% but a low specificity of 42%. These metrics indicate that while the model is effective in detecting stress situations, it faces challenges in correctly identifying animals in thermal comfort, likely due to class imbalance and the need for additional input features to capture environmental adaptability. Consequently, it can be posited that supervised learning models are valuable tools for precision livestock farming, provided that discriminatory limitations are mitigated by refining input characteristics and data balancing. Full article

(This article belongs to the Special Issue Heat Stress in Farm Animals: The Impacts of Climate Change on Animal Comfort Conditions)

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14 pages, 775 KB

Open AccessArticle

Tear Protein Alteration in Dogs with Keratoconjunctivitis Sicca

by Takuya Yogo, Kunihiko Terakado and Kinya Katayama

Animals 2026, 16(2), 160; https://doi.org/10.3390/ani16020160 - 6 Jan 2026

Abstract

Keratoconjunctivitis sicca (KCS) in dogs is an immune-mediated disorder characterized by aqueous tear deficiency, ocular surface inflammation, and risk of vision loss. Although tear quantity is routinely evaluated using the Schirmer tear test (STT), the accompanying qualitative alterations in tear protein composition remain [...] Read more.

Keratoconjunctivitis sicca (KCS) in dogs is an immune-mediated disorder characterized by aqueous tear deficiency, ocular surface inflammation, and risk of vision loss. Although tear quantity is routinely evaluated using the Schirmer tear test (STT), the accompanying qualitative alterations in tear protein composition remain poorly understood. In this exploratory study, we identified and characterized qualitatively differentially expressed tear proteins in samples collected from seven Beagle dogs with KCS and five healthy Beagles. Samples were collected using filter paper, extracted in phosphate-buffered saline, concentrated by trichloroacetic acid precipitation, and then separated via two-dimensional electrophoresis. Differential protein spots were identified by MALDI-TOF-MS-based peptide mass fingerprinting. Total protein concentrations were determined by measuring UV absorbance at 280 nm and were found to be significantly higher in dogs with KCS (30.7 ± 13.5 mg/mL) than in healthy dogs (11.5 ± 1.8 mg/mL, p < 0.05). Five proteins were identified as differentially expressed: serum albumin, lactotransferrin isoform 1, immunoglobulin gamma heavy chain C, major allergen Can f 1, and lysozyme C. High-molecular-weight proteins were upregulated in KCS, whereas low-molecular-weight proteins (<10 kDa, proline-rich protein-like components) were markedly reduced or absent. These compositional shifts suggest that KCS alters both the quantity and qualitative integrity of the tear proteosome, reflecting impaired tear film homeostasis and diminished ocular surface defense. The results support the potential utility of the tear proteome as a source of diagnostic and therapeutic biomarkers in canine KCS. Full article

(This article belongs to the Special Issue Advanced Biomarker Research in Animal Pathological States: Second Edition)

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18 pages, 502 KB

Open AccessReview

Functional Role and Diagnostic Potential of Biomarkers in the Early Detection of Mastitis in Dairy Cows

by Eleonora Dall’Olio, Melania Andrani, Mario Baratta, Fabio De Rensis and Roberta Saleri

Animals 2026, 16(2), 159; https://doi.org/10.3390/ani16020159 - 6 Jan 2026

Abstract

Mastitis remains a prevalent and economically detrimental disease within the dairy industry, profoundly affecting animal welfare, milk quality, and overall production output. Nowadays, Somatic Cell Count (SCC) is widely recognized as the gold-standard indicator for the detection of mastitis; however, its limitations in [...] Read more.

Mastitis remains a prevalent and economically detrimental disease within the dairy industry, profoundly affecting animal welfare, milk quality, and overall production output. Nowadays, Somatic Cell Count (SCC) is widely recognized as the gold-standard indicator for the detection of mastitis; however, its limitations in pathogens discrimination and the lack of early-stage characterization of mastitis highlight the need for complementary diagnostic approaches. This review synthesizes recent research into the development and validation of novel biomarkers for the early and accurate identification of mastitis in dairy cows. The investigation encompasses a range of biological molecules for improving mastitis diagnosis. Biomarkers such as lactoferrin (LTF), β-defensin 4 (DEFB4), vitronectin, paraoxonase 1 (PON1), and N-acetyl-β-D-glucosaminidase (NAGase) show promise in distinguishing between cows not susceptible and cows susceptible to mastitis. Concurrently, nucleic acid-based biomarkers are emerging as a particularly promising frontier. While mitochondrial DNA (mtDNA) has demonstrated insufficient specificity, microRNAs (miRNAs) are gaining attention as highly stable and sensitive indicators of intramammary inflammation, potentially enabling the detection of subclinical infections before they become clinically apparent. Despite these advances, significant challenges related to specificity, reliability, and cost-effectiveness currently hinder the widespread practical application of any single biomarker. Therefore, future research should be directed towards the validation of a synergistic panel of multiple biomarkers to improve mastitis management in dairy cow farms. Full article

(This article belongs to the Special Issue Postpartum Dairy Cow Mammary Health and Mastitis Prevention and Control)

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17 pages, 5366 KB

Open AccessArticle

The Effect of Training on the Expression of Protein and Metabolites in the Plasma Exosomes of the Yili Horse

by Xinxin Yuan, Xinkui Yao, Yaqi Zeng, Jianwen Wang, Wanlu Ren, Tongliang Wang, Xueyan Li, Lipin Yang, Xixi Yang and Jun Meng

Animals 2026, 16(2), 158; https://doi.org/10.3390/ani16020158 - 6 Jan 2026

Abstract

Objective: The effects of physical exercise on protein expression and metabolites in the plasma exosomes of Yili horses are not yet clear. This study aims to elucidate the effects of exercise training on the expression of plasma extracellular vesicle proteins and metabolites in [...] Read more.

Objective: The effects of physical exercise on protein expression and metabolites in the plasma exosomes of Yili horses are not yet clear. This study aims to elucidate the effects of exercise training on the expression of plasma extracellular vesicle proteins and metabolites in Yili horses through multi-omics analysis, providing reference indicators for the training and performance evaluation of Yili horses. Methods: Eight healthy 3-year-old Yili horses were selected for the study, with four untrained horses as the control group and four trained horses as the training group. After conducting proteomic and metabolomic analyses of the plasma exosomes of each group of horses, the differences in protein expression and metabolites were analyzed. Results: Compared with the control group, training significantly increased the levels of proteins in the plasma exosomes of Yili horses that promote muscle function and repair and regulate metabolism and immune function. In addition, carbohydrates and their metabolites were significantly reduced. Conclusion: Training significantly altered the protein composition and metabolites of Yili horse exosomes. These changes mainly provide new insights for the training and evaluation of Yili horse exercise ability, but further research is still needed. Full article

(This article belongs to the Special Issue Unveiling Equine Exercise Physiology for Peak Performance and Optimal Health)

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13 pages, 313 KB

Open AccessFeature PaperArticle

Comparison of N-Terminal Pro-B-Type Natriuretic Peptide Between Cats with Cardiogenic Arterial Thromboembolism and Cats with Occult Cardiomyopathy Without Arterial Thromboembolism

by Michelle A. Oranges, Lisa M. Freeman, Elizabeth A. Rozanski, Emily T. Karlin and John E. Rush

Animals 2026, 16(2), 157; https://doi.org/10.3390/ani16020157 - 6 Jan 2026

Abstract

To help assess the relationship between N-terminal pro-B-type natriuretic peptide (NT-proBNP) and feline cardiogenic arterial thromboembolism (ATE), the objective of this retrospective study was to compare plasma NT-proBNP concentrations between cats with cardiomyopathy that developed ATE (ATE group) and cats with occult cardiomyopathy [...] Read more.

To help assess the relationship between N-terminal pro-B-type natriuretic peptide (NT-proBNP) and feline cardiogenic arterial thromboembolism (ATE), the objective of this retrospective study was to compare plasma NT-proBNP concentrations between cats with cardiomyopathy that developed ATE (ATE group) and cats with occult cardiomyopathy that did not develop ATE within 1 year of testing (occult cardiomyopathy [OCM] group). Cats with cardiomyopathy and congestive heart failure (CHF) but no ATE (CHF group) were included for comparison. Cats with cardiomyopathy that had NT-proBNP testing were classified into ATE, OCM, or CHF groups. Clinical, echocardiographic, treatment, and NT-proBNP data from medical records were reviewed and compared among groups. A receiver operating characteristic curve was generated to develop a cutoff point for NT-proBNP. Cats were then compared based on this cutoff point. The ATE group (n = 25) had significantly higher NT-proBNP concentrations than the OCM group (n = 31; p < 0.001); there was no significant difference in NT-proBNP concentrations between the ATE and CHF groups (p = 0.92). The estimated optimal NT-proBNP cutoff point to separate OCM and ATE groups was 491 pmol/L (sensitivity = 96.0%, specificity = 93.5%). Cats with NT-proBNP > 491 pmol/L had a larger left atrium, thicker left ventricle, lower fractional shortening, and higher prevalence of spontaneous echogenic contrast and left atrial thrombi on echocardiography. These preliminary, hypothesis-generating findings suggest that NT-proBNP concentrations > 491 pmol/L may help detect cats with OCM at risk for ATE, but given the limitations of this retrospective study, prospective studies are needed to evaluate the potential utility of this measurement. Full article

(This article belongs to the Special Issue Clinical Examinations to Promote Diagnosis and Pathophysiological Understanding of Cardiac Disease in Small Animals)

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21 pages, 7464 KB

Open AccessArticle

Enhanced CenterTrack for Robust Underwater Multi-Fish Tracking

by Jinfeng Wang, Mingrun Lin, Zhipeng Cheng, Renyou Yang and Qiong Huang

Animals 2026, 16(2), 156; https://doi.org/10.3390/ani16020156 - 6 Jan 2026

Abstract

Accurate monitoring of fish movement is essential for understanding behavioral patterns and group dynamics in aquaculture systems. Underwater scenes—characterized by dense populations, frequent occlusions, non-rigid body motion, and visually similar appearances—present substantial challenges for conventional multi-object tracking methods. We propose an improved CenterTrack-based [...] Read more.

Accurate monitoring of fish movement is essential for understanding behavioral patterns and group dynamics in aquaculture systems. Underwater scenes—characterized by dense populations, frequent occlusions, non-rigid body motion, and visually similar appearances—present substantial challenges for conventional multi-object tracking methods. We propose an improved CenterTrack-based framework tailored for multi-fish tracking in such environments. The framework integrates three complementary components: a multi-branch feature extractor that enhances discrimination among visually similar individuals, occlusion-aware output heads that estimate visibility states, and a three-stage cascade association module that improves trajectory continuity under abrupt motion and occlusions. To support systematic evaluation, we introduce a self-built dataset named Multi-Fish 25 (MF25), continuous video sequences of 75 individually annotated fish recorded in aquaculture tanks. The experimental results on MF25 show that the proposed method achieves an IDF1 of 82.5%, MOTA of 85.8%, and IDP of 84.7%. Although this study focuses on tracking performance rather than biological analysis, the produced high-quality trajectories form a solid basis for subsequent behavioral studies. The framework’s modular design and computational efficiency make it suitable for practical, online tracking in aquaculture scenarios. Full article

(This article belongs to the Special Issue Fish Cognition and Behaviour)

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15 pages, 855 KB

Open AccessReview

An Integrative Genetic Strategy for Identifying Causal Genes at Quantitative Trait Loci in Chickens

by Akira Ishikawa

Animals 2026, 16(2), 155; https://doi.org/10.3390/ani16020155 - 6 Jan 2026

Abstract

Background: Identifying causal genes underlying quantitative trait loci (QTLs) remains challenging due to small effect sizes and the prevalence of non-coding variants. Although multi-omics integration frameworks such as eQTL- and epigenomic-based approaches and TWAS have advanced gene prioritization, their application in poultry and [...] Read more.

Background: Identifying causal genes underlying quantitative trait loci (QTLs) remains challenging due to small effect sizes and the prevalence of non-coding variants. Although multi-omics integration frameworks such as eQTL- and epigenomic-based approaches and TWAS have advanced gene prioritization, their application in poultry and livestock is often constrained by limited reference panels and tissue resources. This review introduces a cost-effective F₂-based integrative framework and compares it with existing multi-omics strategies. Methods: The proposed framework combines QTL remapping, transcriptome analysis, haplotype frequency comparison, association analysis, and conditional correlation analysis within a single workflow. Causal analysis and quantitative complementation tests using knockout birds are incorporated to identify causal genes. Results: By reusing the original F₂ population employed for QTL mapping, this approach enables hypothesis-independent gene prioritization without requiring additional fine-mapping crosses. Its effectiveness is demonstrated through comparison with conventional multi-omics methods, and the integration of causal analysis and quantitative complementation testing provides robust genetic evidence for pinpointing causal genes. Conclusions: This F₂-based framework efficiently prioritizes and verifies causal gene candidates directly within the mapping population, offering a cost-effective alternative to multi-omics approaches that require large-scale resources. It is broadly applicable to diverse chicken crosses and readily transferable to other small livestock species and model organisms. Full article

(This article belongs to the Special Issue Advances in Genetic Analysis of Important Traits in Poultry)

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24 pages, 1774 KB

Open AccessArticle

Effects of Aspergillus niger and Its Compound Preparations on Methane Emissions and Gastrointestinal Microbiota in Heat-Stressed Holstein Bulls

by Jiangge Wang, Shuaiqi Fu, Xianghui Yin, Shiqin Sun and Tengyun Gao

Animals 2026, 16(2), 154; https://doi.org/10.3390/ani16020154 - 6 Jan 2026

Abstract

The livestock sector is a major source of methane, a potent greenhouse gas, while heat stress impairs ruminant health. This study evaluated the efficacy of Aspergillus niger (AN) and its compound preparation (CP) as feed additives to mitigate methane emissions and heat stress [...] Read more.

The livestock sector is a major source of methane, a potent greenhouse gas, while heat stress impairs ruminant health. This study evaluated the efficacy of Aspergillus niger (AN) and its compound preparation (CP) as feed additives to mitigate methane emissions and heat stress in Holstein bulls. Twenty-four bulls were assigned to a control group (CON), a group supplemented with 6 g/d of AN, and a group with 20 g/d of CP. Methane emissions were measured using the SF₆ tracer technique. Blood and ruminal fermentation parameters were also analyzed. Results showed that both AN and CP significantly reduced total methane emissions, emissions per unit of body weight, and per unit of dry matter intake compared to CON. Supplemented groups had significantly lower rectal temperatures and higher superoxide dismutase activity, with AN also increasing total antioxidant capacity. AN groups showed increased total volatile fatty acids, acetate, and propionate. Microbiota analysis revealed significant beta-diversity shifts with differential taxon enrichment. In conclusion, Aspergillus niger and its compound preparation effectively reduce enteric methane and alleviate heat stress by boosting antioxidant defenses and modulating rumen function, offering a dual-benefit strategy for sustainable ruminant production. Full article

(This article belongs to the Section Cattle)

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