Animals

18 pages, 9008 KB

Open AccessFeature PaperArticle

Enhanced Muscle Flavor in Male Chinese Mitten Crab (Eriocheir sinensis) Driven by Feed-Induced Reconfiguration of Intestinal Volatile Compounds

by Jin Cen, Bo Liu, Qunlan Zhou, Xiaochuan Zheng, Gangchun Xu, Hongyan Tian, Linghong Miao, Huiming Ding, Yongfeng Zhao and Cunxin Sun

Animals 2025, 15(21), 3101; https://doi.org/10.3390/ani15213101 (registering DOI) - 25 Oct 2025

Abstract

The traditional use of iced trash fish (IF) in Chinese mitten crab (Eriocheir sinensis) aquaculture raises sustainability concerns, but the shift to formulated feeds (FF—a commercial compound feed specifically designed to meet nutritional requirements by blending multiple ingredients and containing a [...] Read more.

The traditional use of iced trash fish (IF) in Chinese mitten crab (Eriocheir sinensis) aquaculture raises sustainability concerns, but the shift to formulated feeds (FF—a commercial compound feed specifically designed to meet nutritional requirements by blending multiple ingredients and containing a balance of nutrients) is often hindered by fears of compromising its prized flavor. This study aimed to comprehensively evaluate whether a commercial formulated feed could effectively replace IF without diminishing flavor quality, hypothesizing that FF would alter the intestinal volatile profile, thereby influencing muscle flavor. Male crabs were fed either IF or FF for eight weeks. Muscle flavor was assessed using sensory evaluation, electronic nose (E-nose), and gas chromatography–ion mobility spectrometry (GC–IMS). Volatile compounds in intestinal chyme were also analyzed by GC–IMS to explore potential transfer mechanisms. The results indicated that crabs fed with FF showed higher sensory scores for sweetness. Additionally, the E-nose analysis revealed a clear separation trend between dietary groups and showed markedly higher sensor response values for aromatic compounds, biogenically derived compounds and Maillard reaction products, sulfur-containing organic compounds, aliphatic hydrocarbons, total volatile organic compounds, alcohols and organic solvents, and alkenes in the FF group compared to the IF group. Thirty-four volatiles were discovered in the muscle. Statistical analysis (independent samples t-test) showed that the FF group exhibited significantly elevated levels of 3-methylbutanal-M, propanal, (E)-2-pentenal, 2,3-pentanedione, and pentan-1-ol-M, whereas the IF group exhibited significantly elevated levels of 2-hexanone, dihydro-2(3H)-furanone, butyl acetate, ethyl 2-methylpropanoate, and phenol (p < 0.05). Fourty-eight volatiles were identified in the intestinal chyme. Propanal and ethyl 2-methylpropanoate were the dominant odor contributors based on correlation network analysis. Strong correlations were identified between the flavor profiles of intestinal chyme and muscle, suggesting a potential transfer or transformation of volatiles. This work provides a scientific basis for optimizing aquafeed formulations to ensure sustainable crab production without sacrificing end-flavor quality. Full article

(This article belongs to the Special Issue Advances in Aquaculture Nutrition for Sustainable Health Management)

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23 pages, 22315 KB

Open AccessArticle

Hierarchical Dual-Model Detection Framework for Spotted Seals Using Deep Learning on UAVs

by Jun Liu, Fengxiang Jin, Min Ji, Liang Qu, Juan Wang and Chen Wang

Animals 2025, 15(21), 3100; https://doi.org/10.3390/ani15213100 (registering DOI) - 25 Oct 2025

Abstract

This study introduces a hierarchical dual-model detection framework for accurately monitoring spotted seals (Phoca largha) in the Liaohe River estuary by using deep learning on Unmanned Aerial Vehicles (UAVs). To address challenges such as weak target features, background interference and limited [...] Read more.

This study introduces a hierarchical dual-model detection framework for accurately monitoring spotted seals (Phoca largha) in the Liaohe River estuary by using deep learning on Unmanned Aerial Vehicles (UAVs). To address challenges such as weak target features, background interference and limited edge computing capacity, this study deploys an optimized FF-YOLOv10 lightweight model on UAVs for rapid target localization, followed by an enhanced PP-YOLOv7 model on ground stations for precise detection. The FF-YOLOv10 model reduces computational complexity by 24.2% and increases inference speed by 33.3%, while the PP-YOLOv7 model achieves 94.2% precision with a 1.9% increase in recall rate. This framework provides an efficient and precise technical solution for the long-term ecological monitoring of marine endangered species, supporting habitat conservation policy formulation and ecosystem health assessments. Full article

(This article belongs to the Section Aquatic Animals)

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24 pages, 11929 KB

Open AccessArticle

Effects of Antimicrobial Peptides on the Growth Performance of Squabs Were Investigated Based on Microbiomics and Non-Targeted Metabolomics

by Lihuan Deng, Yingying Yao, Haiying Li, Qingqing Lu and Run Wu

Animals 2025, 15(21), 3099; https://doi.org/10.3390/ani15213099 (registering DOI) - 25 Oct 2025

Abstract

This study aims to investigate the effects of dietary supplementation with AMPs on the growth performance, antioxidant capacity, and intestinal health of squabs. Furthermore, metagenomic and metabolomic approaches were employed to identify key differential bacterial species and metabolites associated with growth performance, and [...] Read more.

This study aims to investigate the effects of dietary supplementation with AMPs on the growth performance, antioxidant capacity, and intestinal health of squabs. Furthermore, metagenomic and metabolomic approaches were employed to identify key differential bacterial species and metabolites associated with growth performance, and thereby the potential mechanisms underlying the enhancement of squab growth and development by AMPs being elucidated. One hundred and twenty pairs of healthy adult White Carneau pigeons (2 years old) were randomly divided into two groups, the control group (CK, fed with basal diet) and antimicrobial peptide group (AP, fed with basal diet +200 mg/kg antimicrobial peptide), with 10 replicates per group and 6 pairs of breeding pigeons per replicate. The experiment lasted for 53 days, including 7 days of prefeeding, 18 days of incubation and 28 days of feeding. In this study, squabs were weighed at 0 and 28 days of age to evaluate growth performance. At 28 days of age, duodenal contents were collected to assess digestive enzyme activities, while jejunal and liver tissues were harvested to determine antioxidant capacity. Intestinal morphology was examined using tissue samples from the duodenum, jejunum, and ileum. Finally, ileal contents were collected for a comprehensive analysis of microbial composition and metabolite profiles in the two experimental groups, employing high-throughput sequencing and LC-MS/MS techniques. The results showed that body weight, liver total antioxidant capacity (T-AOC), jejunal malondialdehyde (MDA) content, jejunum and ileum villus height-to-crypt depth ratio (VH/CD) were significantly increased, and jejunal crypt depth (CD) was significantly decreased in the AP group at 28 days of age (p < 0.05). In addition, the microbiome data showed that Lactobacillus in the AP group was a biomarker with significant differences (p < 0.05). Metabolomics analysis showed that the steroid hormone biosynthesis pathway was significantly different between the two groups (p < 0.01). In addition, the content of potentially beneficial metabolites (Biotin, beta-Tocotrienol, 7-Chloro-L-tryptophan and Dihydrozeatin) was significantly increased in the AP group (p < 0.05). These results indicate that dietary AMPs can significantly improve the body weights, liver antioxidant capacity and jejunum and ileum VH/CD of squabs. Full article

(This article belongs to the Section Animal Physiology)

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26 pages, 5152 KB

Open AccessArticle

Evaluating the Adjuvant Therapeutic Effects of Probiotic Strains Lactococcus cremoris and Lacticaseibacillus paracasei on Canine Atopic Dermatitis and Their Impact on the Gut and Skin Microbiome

by Hsiao-Wen Huang, Ting-Chen Yeh, Jui-Chun Hsieh, Ching-Wen Tsai, Ya-Jane Lee and Ming-Ju Chen

Animals 2025, 15(21), 3098; https://doi.org/10.3390/ani15213098 (registering DOI) - 24 Oct 2025

Abstract

This study investigated the therapeutic potential of a novel probiotic combination consisting of Lactococcus cremoris subsp. cremoris MP01 and Lacticaseibacillus paracasei subsp. paracasei MP02, collectively referred to as LCP, in the treatment of canine atopic dermatitis (CAD). In a 60-day open-label, single-arm trial [...] Read more.

This study investigated the therapeutic potential of a novel probiotic combination consisting of Lactococcus cremoris subsp. cremoris MP01 and Lacticaseibacillus paracasei subsp. paracasei MP02, collectively referred to as LCP, in the treatment of canine atopic dermatitis (CAD). In a 60-day open-label, single-arm trial involving eight dogs, notable clinical improvements were observed following daily LCP treatment, as evidenced by decreasing trends in Canine Atopic Dermatitis Extent and Severity Index and Pruritus Visual Analogue Scale scores, as well as a significant reduction in serum immunoglobulin E levels (p < 0.05). Microbiome and short-chain fatty acid (SCFA) analyses were subsequently conducted in a representative subset of six dogs to explore the effects of LCP on the fecal and skin microbial ecosystems. Concomitant alterations in gut and skin microbiome were observed, including a significant reduction in abundance of Erysipelotrichaceae (p < 0.05) and non-significant decreasing trends in Romboutsia, Escherichia/Shigella spp., and Shigella flexneri, along with a trend toward increased SCFA production. Functional prediction using PICRUSt suggested potential involvement of immune- and infection-related signaling pathways, including those associated with nucleotide-binding oligomerization domain-like receptors, retinoic acid-inducible gene I-like receptors and Shigellosis, supporting the hypothesis that LCP may exert its effects through modulation of the gut–skin axis. These findings support LCP as a safe and promising adjunct therapy for CAD, offering a novel microbiome-targeted approach targeting both clinical symptoms and underlying dysbiosis. Further investigation is warranted to optimize probiotic formulations and better understand the mechanisms underlying microbiome-mediated immune modulation in canine allergy. Full article

(This article belongs to the Special Issue Nutrition, Physiology and Metabolism of Companion Animals)

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18 pages, 5581 KB

Open AccessArticle

A New Strategy to Identify Naturally Presenting SLA-I Bound Peptides Derived from the O Serotype of Foot-and-Mouth Disease Virus, by Mild Acid Elution in a VP1 Stably Expressed PK15 Cell Line

by Yong-Yu Gao, Zong-Hui Zhang, Chen-Jun Sang, Yong Han, Yu-Die Cao, Yue Tang, Gui-Xue Hu, Zi-Bin Li and Feng-Shan Gao

Animals 2025, 15(21), 3097; https://doi.org/10.3390/ani15213097 (registering DOI) - 24 Oct 2025

Abstract

Multi-epitopes of FMDV can be used to develop a novel vaccine. Determining how to screen naturally presenting epitope peptides derived from FMDV is crucial for advancing progress in this area. In this study, a transient expression plasmid named pEGFP-N1-VP1 was transfected into Porcine [...] Read more.

Multi-epitopes of FMDV can be used to develop a novel vaccine. Determining how to screen naturally presenting epitope peptides derived from FMDV is crucial for advancing progress in this area. In this study, a transient expression plasmid named pEGFP-N1-VP1 was transfected into Porcine Kidney Epithelial cells 15 (PK15). The positive cells that stably expressed the O-VP1 gene of FMDV were screened with gradient concentrations of G418 (Geneticin). The constructed pEGFP-N1-VP1/PK15 cell line was eluted by pH 3.3 phosphate buffer to isolate the eluted peptides, followed by desalting, liquid chromatography–tandem mass spectrometry (LC–MS/MS), a flow cytometric analysis of SLA-I expression, and an ELISA detection of SLA-I bound peptides. It was demonstrated that a PK15 cell line stably expressing the VP1 gene was initially screened out at 500 μg/mL of G418, followed by culturing at 300 μg/mL. The O-VP1 expression was identified using an image analysis system, RT-PCR, and Western blot analysis. Thirty-seven peptides derived from O-VP1 were eluted from the constructed cell line. The flow cytometric analysis and ELISA detection results showed that the eluted peptides were associated with SLA-I and bound. This is the first known study to construct a cell line for screening naturally presenting antigenic peptides derived from the O serotype of FMDV. Full article

(This article belongs to the Section Pigs)

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32 pages, 14182 KB

Open AccessArticle

Effects of Soybean Meal Replacement on Growth Performance, Rumen Fermentation, Rumen Microorganisms, and Metabolites in Dumont Lambs

by Henan Lu, Hairong Wang, Boyang Li, Zenghao Lv, Shufang Li, Yuhao Xia and Lina Wang

Animals 2025, 15(21), 3096; https://doi.org/10.3390/ani15213096 (registering DOI) - 24 Oct 2025

Abstract

This study investigated the effects of replacing part of the soybean meal in the diet of Dumont lambs with urea, rapeseed meal, and cottonseed meal on their growth performance and rumen fermentation and combined rumen microbial metagenomics and metabolomics to explain the reasons [...] Read more.

This study investigated the effects of replacing part of the soybean meal in the diet of Dumont lambs with urea, rapeseed meal, and cottonseed meal on their growth performance and rumen fermentation and combined rumen microbial metagenomics and metabolomics to explain the reasons for the changes in phenotypic data. Twenty-four healthy male Dumont lambs were divided into four groups: soybean meal group (T1, control group), group with 1.5% urea replacing 6.4% soybean meal (T2), group with 1% urea replacing 4.3% soybean meal (T3), and group with 1% urea + 6.6% cottonseed meal +5% rapeseed meal replacing all soybean meal (19%) (T4), following the principle of equal energy and nitrogen. Urea, rapeseed meal, and cottonseed meal have different degradation rates in the rumen, primarily stimulating arginine biosynthesis, sulphur metabolism, and carbon fixation in photosynthetic organisms through Prevotella genus mediation, thereby influencing the accumulation of metabolites such as 9,10-DiHOME, DG (PGJ2/a-15:0/0:0), isonicotinate and taxifolin, affecting rumen fermentation. Compared with the T1 group, the T2 group showed significantly increased ammonia nitrogen (NH₃-N) and microbial protein (MCP) content (p < 0.01) and improved fructose and mannose metabolic capacity (p < 0.05). The T3 group showed a significant increase in total volatile fatty acids (TVFA) and MCP content (p < 0.01), which facilitated the absorption of subsequent nutrients. In the T4 group, different degradation rates of nitrogen resources and rapeseed meal + cottonseed meal contained abundant and complementary amino acids, which improved rumen fermentation, enhanced rumen microbial and metabolite diversity, and optimized the synergistic metabolic efficiency of carbon, nitrogen and sulphur. However, the specific mechanisms of post-rumen metabolism and absorption require further investigation. Full article

(This article belongs to the Section Small Ruminants)

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14 pages, 4062 KB

Open AccessArticle

Mammary Epithelial Cell Spheroid: Stabilization Through Vascular-Wall Mesenchymal Stem Cells and Endothelial Cells Co-Culture

by Debora La Mantia, Roberta Salaroli, Biljana Petrovic, Domenico Ventrella, Augusta Zannoni, Monica Forni and Chiara Bernardini

Animals 2025, 15(21), 3095; https://doi.org/10.3390/ani15213095 (registering DOI) - 24 Oct 2025

Abstract

Over the past decade, the interest in using 3D cell culture models for studying the mammary gland in biomedical and veterinary fields has increased, but a fully functional in vitro model for domestic species is still lacking. Multiple cellular components, including epithelial cells, [...] Read more.

Over the past decade, the interest in using 3D cell culture models for studying the mammary gland in biomedical and veterinary fields has increased, but a fully functional in vitro model for domestic species is still lacking. Multiple cellular components, including epithelial cells, vascular endothelial cells, and stromal/stem cells, sustain the secretory mammary gland tissue in a well-organized 3D architecture. Considering the Göttingen Minipigs widely used for translational lactation studies, this work aimed to establish a 3D culture protocol to generate mammary heterogeneous multicellular spheroids composed of three different Göttingen Minipigs primary cells: mammary epithelial cells (mpMECs), aortic endothelial cells (mpAECs), and vascular-wall mesenchymal stem cells (mpVW-MSCs). Cells were cultured with hanging-drop (HD) and ultra-low-adherence plate (ULA) methods, evaluating aggregate formation in both monocultures and co/triple co-cultures. Brightfield area, eccentricity, viability, and cell distribution were analyzed. Results showed mpMECs formed irregular aggregates in both HD and ULA, while more compact and viable spheroids were formed when co-cultured with mpVW-MSCs and mpAECs by ULA. A well-organized cellular distribution was demonstrated by cytokeratin-18, vimentin, and e-NOS immunofluorescence analysis. In conclusion, this study established a stable 3D mammary multicellular spheroid model, representing a promising tool for future studies on hormonal modulation and mammary gland physiology. Full article

(This article belongs to the Collection Stem Cells in Domestic Animals: Applications in Health and Production)

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18 pages, 1090 KB

Open AccessArticle

Dynamic Changes in Amino Acid Release Patterns of Different Plant Protein Sources During In Vitro Digestion and Their Nutritional Value Assessment

by Yueli Fan, Zehua Kou, Jiahua Cao, Zhongshen Wang, Tianrui Zhang, Rui Han and Dongsheng Che

Animals 2025, 15(21), 3094; https://doi.org/10.3390/ani15213094 (registering DOI) - 24 Oct 2025

Abstract

A gastric–intestinal two-step enzymatic hydrolysis in vitro digestion simulation system was used to systematically investigate the digestion kinetics and amino acid release characteristics of five plant protein sources: soybean meal, rapeseed meal, corn DDGS, corn gluten meal, and corn germ meal. The results [...] Read more.

A gastric–intestinal two-step enzymatic hydrolysis in vitro digestion simulation system was used to systematically investigate the digestion kinetics and amino acid release characteristics of five plant protein sources: soybean meal, rapeseed meal, corn DDGS, corn gluten meal, and corn germ meal. The results showed that in the gastric digestion phase (120 min), the protein hydrolysis degree of soybean meal was the highest (61.8%, p < 0.001), which was 4.4 times that of corn gluten meal (14.0%). In the intestinal digestion phase (240 min), the low-molecular-weight peptide release of corn gluten meal (31.2 mg/g) was significantly higher than that of corn DDGS (17.4 mg/g), showing a “weak in the stomach but strong in the intestine” characteristic. The “nutritional value equivalence” model constructed with soybean meal as the reference showed that the gastric digestion phase equivalence of rapeseed meal was only 32.2% (significantly lower than other materials), and the intestinal digestion phase equivalence of corn gluten meal was 62.9%. This study clarified the differences in digestion characteristics and key related indicators of different plant protein sources, providing quantitative references and scientific support for the food and feed industries to precisely select protein sources according to digestion phases and optimize the formula design. Full article

(This article belongs to the Special Issue Alternative Protein Sources for Animal Feeds)

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17 pages, 695 KB

Open AccessReview

Passive Immunity Establishment Through Colostral IgG Absorption in Neonatal Ruminants: Foundation for Efficient Ruminant Production

by Chao Yang, Mei Du, Anum Ali Ahmad, Yan Cheng and Kefyalew Gebeyew

Animals 2025, 15(21), 3093; https://doi.org/10.3390/ani15213093 (registering DOI) - 24 Oct 2025

Abstract

Passive immunity, the acquisition of specific immune protection through external antibodies or immune components, is critically important for neonatal survival. In ruminants, however, neonatal hypogammaglobulinemia, a consequence of their epitheliochorial placental structure preventing prenatal antibody transfer, often leads to high morbidity and mortality. [...] Read more.

Passive immunity, the acquisition of specific immune protection through external antibodies or immune components, is critically important for neonatal survival. In ruminants, however, neonatal hypogammaglobulinemia, a consequence of their epitheliochorial placental structure preventing prenatal antibody transfer, often leads to high morbidity and mortality. Consequently, neonatal ruminants are entirely dependent on the timely consumption of colostrum to acquire sufficient immunoglobulin G (IgG) for protection. Establishing robust passive immunity is therefore a cornerstone for their survival, healthy development, and future production efficiency. This review synthesizes current knowledge on the establishment of passive immunity in neonatal ruminants. We first outline the fundamental principles of passive immunity transfer, then delve into the specific pathways and molecular mechanisms in ruminants. Key factors influencing this process are subsequently discussed. Furthermore, we highlight the long-term impact of passive immunity on adult production performance. This review aims to provide a scientific foundation for optimizing colostrum management strategies and to stimulate future research into the intricate mechanisms of IgG absorption. Full article

(This article belongs to the Special Issue Advanced Welfare-Oriented, Efficient, and High-Quality Ruminant Production: Mechanisms and Practices)

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32 pages, 758 KB

Open AccessFeature PaperReview

Alpaca (Vicugna pacos) Husbandry and Their Welfare

by Renata Pilarczyk, Małgorzata Bąkowska, Bogumiła Pilarczyk, Agnieszka Tomza-Marciniak, Beata Seremak, Jan Udała, Paulius Matusevičius and Ramutė Mišeikienė

Animals 2025, 15(21), 3092; https://doi.org/10.3390/ani15213092 (registering DOI) - 24 Oct 2025

Abstract

A key diagnostic tool in breeding practice is systematic observation: by identifying abnormalities in natural behaviour, it can highlight early signs of reduced welfare or physical and mental health issues in livestock and other animals. The aim of this paper is to review [...] Read more.

A key diagnostic tool in breeding practice is systematic observation: by identifying abnormalities in natural behaviour, it can highlight early signs of reduced welfare or physical and mental health issues in livestock and other animals. The aim of this paper is to review current literature to provide a detailed analysis of the factors affecting the physical and mental health of alpacas (Vicugna pacos) with regard to their husbandry method. It analyses various behavioural patterns such as stress reactions to strangers, isolation and grooming, as well as social interactions (hierarchy, bonding, affiliative and aggressive behaviour), reproductive activity (courtship, copulation, partner rejection), and resting and foraging rhythms. It also discusses the influence of socialisation on human–animal relationships and the role of the environment, and its enrichment, in proper behavioural functioning. It also examines the significant role played by nutrition and the environment in ensuring alpaca welfare, and how it can be maintained during grooming and training. It pays special attention to the use of behavioural tests to assess the level of trust and tameness in alpacas. Systematic behaviour observation allows a quick response to deteriorating welfare, thus facilitating more efficient herd management and reducing the risk of more serious health and behavioural problems. Full article

(This article belongs to the Section Animal Welfare)

14 pages, 3193 KB

Open AccessFeature PaperArticle

Comparative Chromosome Painting Clarifies the Intraspecific Chromosomal Variation in Two Ctenomys Species (Rodentia: Ctenomyidae)

by Thays Duarte de Oliveira, Natasha Ávila Bertocchi, Luciano Cesar Pozzobon, Ivanete de Oliveira Furo, Edivaldo Herculano Corrêa de Oliveira, Jorge C. Pereira, Malcolm A. Ferguson-Smith, Rafael Kretschmer and Thales R. O. de Freitas

Animals 2025, 15(21), 3091; https://doi.org/10.3390/ani15213091 (registering DOI) - 24 Oct 2025

Abstract

Background: Ctenomys is a subterranean rodent genus known for exhibiting the highest levels of chromosome variation, both among species (2n = 10 to 70) and within species. Ctenomys minutus is particularly notable for its extensive chromosomal diversity, comprising the greatest number of [...] Read more.

Background: Ctenomys is a subterranean rodent genus known for exhibiting the highest levels of chromosome variation, both among species (2n = 10 to 70) and within species. Ctenomys minutus is particularly notable for its extensive chromosomal diversity, comprising the greatest number of described cytotypes within this genus. In contrast, Ctenomys lami presents the highest degree of karyotypic variation within a comparatively restricted geographic range. Both species inhabit the coastal plain of southern Brazil: C. minutus occurs in dunes and sandy fields extending from Laguna (Santa Catarina State) to São José do Norte (Rio Grande do Sul State), whereas C. lami is restricted to the “Coxilha das Lombas” region, which lies parallel to the distribution of C. minutus in Rio Grande do Sul State. Despite their close evolutionary relationship and the absence of external morphological differences, the mechanism underlying their karyotypic divergence remains poorly understood. Methods: In this study, we applied whole-chromosome painting using probes from Ctenomys flamarioni to investigate chromosomal evolution in C. minutus and C. lami. Results: The resulting homology maps revealed a variety of chromosomal rearrangements that differentiate cytotypes both within and between these species. Comparative analyses demonstrated substantial karyotypic divergence from C. flamarioni, although some entire chromosomes and large chromosomal segments remained conserved between C. minutus and C. lami. Our findings underscore the critical role of chromosomal rearrangements in shaping the diversification of Ctenomys. Additionally, we identified shared chromosomal rearrangements in C. minutus and C. lami, which are likely restricted to the torquatus group. Conclusions: These rearrangements provide new insights into the processes driving chromosomal evolution in genus Ctenomys. Full article

(This article belongs to the Special Issue Vertebrate Cytogenomics: Insights into Genome and Chromosome Evolution)

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18 pages, 315 KB

Open AccessArticle

Enhanced Farrowing Efficiency and Sow Performance with Escherichia coli-Derived 6-Phytase Supplementation During Late Gestation and Lactation

by Débora Cristina Peretti, Marco Aurélio Callegari, Cleandro Pazinato Dias, Gabrieli de Souza Romano Bergamo, Bindhu Lakshmibai Vasanthakumari, Mara Cristina Ribeiro da Costa, Rafael Humberto de Carvalho and Caio Abércio da Silva

Animals 2025, 15(21), 3090; https://doi.org/10.3390/ani15213090 (registering DOI) - 24 Oct 2025

Abstract

Phytase releases phosphorus from phytate and may confer extra-phosphoric benefits in sows. We tested whether Escherichia coli-derived 6-phytase during late gestation and lactation improves sow and litter outcomes. In a randomized complete block trial, 186 TN70 sows received a phytase-free positive control [...] Read more.

Phytase releases phosphorus from phytate and may confer extra-phosphoric benefits in sows. We tested whether Escherichia coli-derived 6-phytase during late gestation and lactation improves sow and litter outcomes. In a randomized complete block trial, 186 TN70 sows received a phytase-free positive control (adequate Ca and available P) or Ca- and P-reduced diets with 500, 1500, or 2500 FTU/kg. Outcomes included sow body condition, lactation feed intake and feed conversion ratio (FCR), farrowing duration and blood glucose, piglet weaning performance and diarrhea scores, maternal serum Ca and P (farrowing, weaning), and piglet glutathione peroxidase (GPx) and superoxide dismutase (SOD; day 14). Phytase increased lactation intake by 4.4–5.6%; farrowing duration was shorter at all doses (−24.2, −23.8, and −14.8 min; up to −8.1%). Litter weaning weight rose by 6.1–8.2%, and piglet average daily gain increased by 9.1% at 2500 FTU/kg. Maternal Ca and P increased dose-responsively, especially at weaning (Ca +73% at 500–1500 FTU/kg; +140% at 2500; P +55%, +59%, +118%). Diarrhea counts declined at selected doses (e.g., scores 1–2: −17% at 500 FTU/kg), and piglet SOD decreased with dose (−8.6% to −39.3%); GPx showed modest modulation. Sow body weight, backfat, and the weaning-to-estrus interval were unchanged. In Ca- and P-reduced diets, conventional and super-dosed phytase enhanced mineral bioavailability and peripartum efficiency, supporting heavier litters without compromising sow condition. Full article

(This article belongs to the Section Animal Nutrition)

11 pages, 2347 KB

Open AccessCase Report

Use of Contrast-Enhanced Ultrasound in Suspected Traumatic or Spontaneous Renal Injury in Cats: A Case Series

by Simone Perfetti, Carolina Gai, Nikolina Linta, Giacomo Tamburini, Erika Monari, Elena Ciuffoli and Alessia Diana

Animals 2025, 15(21), 3089; https://doi.org/10.3390/ani15213089 (registering DOI) - 24 Oct 2025

Abstract

Contrast-enhanced ultrasound (CEUS) is increasingly applied in veterinary medicine as a safe, rapid, and non-invasive imaging technique for assessing renal disorders. Despite its expanding use, the literature on its application in feline renal trauma remains scarce. This retrospective study aimed to describe CEUS [...] Read more.

Contrast-enhanced ultrasound (CEUS) is increasingly applied in veterinary medicine as a safe, rapid, and non-invasive imaging technique for assessing renal disorders. Despite its expanding use, the literature on its application in feline renal trauma remains scarce. This retrospective study aimed to describe CEUS findings in cats with suspected traumatic renal injuries. Medical records were reviewed for cats that underwent both B-mode ultrasonography and CEUS, with findings confirmed by follow-up, surgery, or cytology. Three cats met the inclusion criteria. Two presented focal or multifocal renal lesions ranging from 10 to 20 mm in diameter, with heterogeneous echotexture, distortion of renal contours, and non-enhancing areas on CEUS consistent with hematomas or lacerations. The third cat showed a circumferential subcapsular halo (approximately 3–5 mm thick) with evidence of contrast leakage, compatible with limited active hemorrhage. CEUS appeared effective in identifying and characterizing renal injuries, offering valuable information to support clinical decision-making and guide both conservative and surgical management. Nevertheless, due to the limited sample size and the absence of quantitative data, these results should be considered preliminary. Further prospective studies are warranted to confirm the diagnostic performance and clinical utility of CEUS in feline renal trauma. Full article

(This article belongs to the Special Issue Advances in Canine and Feline Nephrology and Urology)

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10 pages, 1857 KB

Open AccessCommunication

The First Record of Taiga Shrew in Lithuania

by Linas Balčiauskas

Animals 2025, 15(21), 3088; https://doi.org/10.3390/ani15213088 (registering DOI) - 24 Oct 2025

Abstract

The taiga shrew (Sorex isodon), a boreal forest species widely distributed across northern Eurasia, had not previously been recorded in the Baltic states. We report the first occurrence of S. isodon in Lithuania, which extends the species range westward into the [...] Read more.

The taiga shrew (Sorex isodon), a boreal forest species widely distributed across northern Eurasia, had not previously been recorded in the Baltic states. We report the first occurrence of S. isodon in Lithuania, which extends the species range westward into the Baltic region. The specimen, which was initially misidentified as Sorex araneus, was collected in 2023 near Vilnius during non-systematic snap-trapping. A detailed examination of tail, cranial, and dental characteristics confirmed the identification as S. isodon. The individual was a non-adult male, weighing 5.8 g, and was trapped in a wet mixed forest influenced by beaver activity. This suggests that the microhabitat conditions are similar to those reported in Belarus and Ukraine. This finding extends the western boundary of S. isodon distribution by approximately 200–630 km. This study underscores the potential underreporting of this species in the Baltic region and highlights the importance of revisiting existing skull collections, conducting targeted trapping, and performing genetic analyses. Verifying additional records will clarify the habitat preferences and conservation relevance of S. isodon at the western edge of its range. Full article

(This article belongs to the Section Wildlife)

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3 pages, 158 KB

Open AccessEditorial

Happiness Is in the Field

by Dominique-Marie Votion

Animals 2025, 15(21), 3087; https://doi.org/10.3390/ani15213087 (registering DOI) - 24 Oct 2025

Abstract

Common sense suggests that ‘happiness is in the field’, as grasslands are considered environments that promote the health and well-being of grazing animals [...] Full article

(This article belongs to the Special Issue Pasture-Associated Poisoning in Grazing Animals)

17 pages, 5369 KB

Open AccessArticle

Improved Eggshell Quality in Aged Hens Through Circadian Gut Microbiota and Metabolite Changes Induced by a 28-h Ahemeral Light Cycle

by Junjie Xu, Xinxin Li, Xuelu Liu, Xinling Wu, Yihao Fan, Yichun Yao, Rongcai Zhang, Dehe Wang, Yifan Chen, Erying Hao, Yanyan Sun, Jilan Chen, Hui Chen and Lei Shi

Animals 2025, 15(21), 3086; https://doi.org/10.3390/ani15213086 (registering DOI) - 24 Oct 2025

Abstract

The decline in eggshell quality of aged laying hens represents a major economic challenge in poultry production. While a 28 h ahemeral light cycle has been shown to improve eggshell quality, its underlying mechanism remains unclear. This study randomly assigned 260 74-week-old Hy-Line [...] Read more.

The decline in eggshell quality of aged laying hens represents a major economic challenge in poultry production. While a 28 h ahemeral light cycle has been shown to improve eggshell quality, its underlying mechanism remains unclear. This study randomly assigned 260 74-week-old Hy-Line Brown laying hens to two light cycle groups, a normal 24 h cycle group (16L:8D) and a 28 h ahemeral cycle group (16L:12D). Each treatment comprised 130 hens divided into two replicate groups. The trial lasted 16 weeks. We systematically analyzed circadian rhythms of gut microbiota and serum metabolites using 16S rRNA sequencing and untargeted metabolomics. Compared with the 24 h cycle, the 28 h cycle significantly enhanced eggshell thickness by 0.04 mm and 0.02 mm, and eggshell strength by 4.19 N and 4.76 N at 79 and 84 wk, respectively. Mechanistically, the 28 h light cycle remodeled the circadian rhythms of gut microbiota, increasing their richness and diversity, and altered the rhythmic patterns of serum metabolites. We identified nine microbial genera and three hundred seventy metabolites that exhibited opposite rhythmic patterns under the two light cycles. These changes were primarily enriched in pathways related to amino acid, carbohydrate, lipid, and energy metabolism. Correlation analysis further revealed strong associations between key microbes and functional metabolites. Weissella promotes calcium deposition in eggshells through synergistic interactions with calcium chelators such as gluconic acid and threonine acid. Meanwhile, YRC22 and Paludibacter synergistically support membrane formation substances, thereby promoting the proliferation of uterine epithelial cells and eggshell formation. Our findings indicate that the 28 h ahemeral light cycle improved eggshell quality in aged hens by remodeling the circadian rhythms of gut microbiota and metabolites, thereby synergistically enhancing calcium ion absorption and uterine tissue health. This provides a novel theoretical basis and practical direction for improving late-phase egg quality through light management strategies. Full article

(This article belongs to the Section Poultry)

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13 pages, 1352 KB

Open AccessArticle

Microvillus in LBW Meishan Piglets Preserved Microvillus Integrity Alongside Impaired Intestinal Barrier Function in Low-Birth-Weight Meishan Neonatal Piglets

by Li Dong, You Wu, Zhixuan Sun, Hongrong Wang and Lihuai Yu

Animals 2025, 15(21), 3085; https://doi.org/10.3390/ani15213085 (registering DOI) - 24 Oct 2025

Abstract

Despite lower birth weight, Meishan piglets exhibit a notably higher pre-weaning survival rate compared to Western commercial breeds. This study aimed to evaluate the effect of low birth weight (LBW) on intestinal barrier function in Meishan neonates. Six pairs of neonatal piglets (one [...] Read more.

Despite lower birth weight, Meishan piglets exhibit a notably higher pre-weaning survival rate compared to Western commercial breeds. This study aimed to evaluate the effect of low birth weight (LBW) on intestinal barrier function in Meishan neonates. Six pairs of neonatal piglets (one normal birth weight, NBW: 0.85 ± 0.06 kg; one LBW: 0.65 ± 0.02 kg) from the same sow were euthanized at birth prior to suckling. Morphological parameters, goblet cell density, antioxidant enzyme activities, cytokine gene expression, and tight junction protein levels in the small intestine (SI) were assessed. Results showed that LBW piglets had a significantly higher SI length-to-body weight ratio (p < 0.05), along with reduced villus height, villus height-to-crypt depth ratio, and villus surface area in the jejunum and ileum (p < 0.01). Notably, microvillus structure remained intact despite the presence of mitochondrial swelling. LBW piglets also exhibited decreased goblet cell numbers, lower antioxidant capacity, dysregulated expression of cytokines (CD8, IFNγ, IL4, IL2), and reduced levels of mucin 2, ZO-1, and occludin (p < 0.05). In conclusion, although LBW Meishan piglets showed impairments in multiple aspects of intestinal barrier function, the structural integrity of the microvillus was preserved, which may contribute to their higher survival rate and represents a key adaptive advantage over commercial pig breeds. Full article

(This article belongs to the Section Pigs)

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11 pages, 371 KB

Open AccessArticle

A One Health Perspective on Heartworm Disease: Allergy Risk in Owners of Infected Dogs in Gran Canaria (Spain)

by José Alberto Montoya-Alonso, Alfonso Balmori-de la Puente, Noelia Costa-Rodríguez, Jorge Isidoro Matos, Elena Carretón and Rodrigo Morchón

Animals 2025, 15(21), 3084; https://doi.org/10.3390/ani15213084 - 24 Oct 2025

Abstract

Heartworm disease, caused by Dirofilaria immitis and transmitted by mosquitoes, represents a major veterinary and zoonotic concern. Beyond its recognized cardiopulmonary effects in dogs, increasing evidence suggests that parasite-derived antigens may contribute to allergic disease in humans. Gran Canaria (Canary Islands, Spain) is [...] Read more.

Heartworm disease, caused by Dirofilaria immitis and transmitted by mosquitoes, represents a major veterinary and zoonotic concern. Beyond its recognized cardiopulmonary effects in dogs, increasing evidence suggests that parasite-derived antigens may contribute to allergic disease in humans. Gran Canaria (Canary Islands, Spain) is a hyperendemic region for heartworm disease, offering a unique setting to explore this potential relationship. Therefore, the aim was to analyze the association between canine infection with D. immitis and the occurrence of allergic conditions in their owners on Gran Canaria. This cross-sectional study included 644 dogs and their respective owners. Dogs were tested for circulating D. immitis antigens using a commercial rapid test. Owners provided information on allergy through medical anamnesis. Associations between canine infection status, animal-related variables (age, sex, breed, habitat), and owner-reported allergies (age, sex) were analyzed using chi-square and Fisher’s exact tests with Bonferroni correction. The results showed that 46.4% of dogs tested positive for D. immitis and 43.8% of owners reported allergic diseases. However, 24.64% of dog owners who tested negative for heartworm disease and 65.89% of owners with positive pets reported allergic diseases. In fact, owners of infected dogs exhibited a significantly higher prevalence of allergies compared to those of uninfected dogs. Canine age, sex, breed, and owner demographics were not associated with infection or allergies. However, dog habitat was significantly associated with both outcomes: dogs living indoors had lower infection rates, while owners of infected dogs in mixed indoor–outdoor environments reported the highest rates of allergy. This study provides the first epidemiological evidence of an association between canine heartworm infection and allergies in owners within a hyperendemic European region. The findings suggest that cohabitation with infected dogs may represent a novel risk factor for atopy, reinforcing the importance of integrated preventive measures and highlighting the relevance of a One Health approach. Full article

(This article belongs to the Section Veterinary Clinical Studies)

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18 pages, 9796 KB

Open AccessFeature PaperArticle

Integrative Transcriptomic and Proteomic Analysis Reveals CaMK4-Mediated Regulation of Proliferation in Goat Skeletal Muscle Satellite Cells

by He Cong, Lu Xu, Yaolong Liu, Zixuan Wang, Tao Ren, Pengcheng Ruan, Haoyuan Zhang, Chengli Liu, Yanguo Han, Pengfei Hu, Yan Zeng, Simone Ceccobelli and Guangxin E

Animals 2025, 15(21), 3083; https://doi.org/10.3390/ani15213083 - 24 Oct 2025

Abstract

CaMK4, a calcium/calmodulin-dependent protein kinase, is an important mediator of cellular signal transduction, yet its role in the regulation of skeletal muscle satellite cells (MuSCs) in goats has remained unclear. In this study, CaMK4 overexpression and knockdown models were established, and integrated [...] Read more.

CaMK4, a calcium/calmodulin-dependent protein kinase, is an important mediator of cellular signal transduction, yet its role in the regulation of skeletal muscle satellite cells (MuSCs) in goats has remained unclear. In this study, CaMK4 overexpression and knockdown models were established, and integrated transcriptomic and proteomic analyses were performed to systematically elucidate its regulatory network. CaMK4 overexpression altered key pathways associated with cell proliferation and muscle development, including cAMP, PI3K-Akt, and actin cytoskeleton regulation, while proteomic data highlighted calcium signaling and JAK-STAT pathways. Conversely, CaMK4 knockdown enhanced MuSC proliferation by upregulating cell cycle-related genes and proteins. Integrated analyses further identified that Galectin-9 (LGALS9), Collagen triple helix repeat containing-1 (CTHRC1), Hyaluronan Synthase 1 (HAS1), and L-Threonine Dehydrogenase (TDH) may serve as potential key nodes regulating cell cycle, apoptosis, and metabolic control. This suggests a regulatory role for CaMK4. Collectively, these findings provide a mechanistic framework for understanding CaMK4 function in ruminant muscle development and may offer insights for improving goat muscle growth, meat quality traits, and production efficiency. Full article

(This article belongs to the Special Issue Genetics and Breeding for Enhancing Production Traits in Ruminants)

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20 pages, 3828 KB

Open AccessArticle

Identification of Expression Quantitative Trait Loci (eQTL) for Adipose-Specific Regulatory Mechanisms in Hanwoo (Korean Cattle)

by Junyoung Lee, Taejoon Jeong, Woncheoul Park, Sunsik Jang, Poong-Yeon Lee and Dajeong Lim

Animals 2025, 15(21), 3082; https://doi.org/10.3390/ani15213082 - 24 Oct 2025

Abstract

Understanding the genetic regulatory mechanisms of fat accumulation is crucial for improving beef quality. Hanwoo (Korean native cattle) is renowned for its high intramuscular fat (marbling), yet the genetic regulation of adipose gene expression remains insufficiently understood. In this study, we performed expression [...] Read more.

Understanding the genetic regulatory mechanisms of fat accumulation is crucial for improving beef quality. Hanwoo (Korean native cattle) is renowned for its high intramuscular fat (marbling), yet the genetic regulation of adipose gene expression remains insufficiently understood. In this study, we performed expression quantitative trait loci (eQTL) analysis using RNA-Seq data and genotype data from backfat tissue of 75 Hanwoo steers to identify regulatory variants associated with adipose deposition. A total of 25,042 significant cis-eQTL associations (FDR < 0.05) were identified, and 5362 unique top cis-eQTL pairs were retained after gene-wise filtering. Key cis-regulated genes included AGBL1, CACNG1, MYO18B, and DUSP29, which are involved in cytoskeletal organization, muscle development and calcium signaling. Three major cis-regulatory hotspots were located on BTA15 (BTA15:50354741) and BTA21 (BTA21:21526143, and BTA21:21541921). Permutation-based analysis (100,000 iterations) was conducted to control false positives, identifying 12 statistically significant trans-eQTL hotspots (FDR q < 0.05), of which SNP 6:60512276 and SNP 21:17035557 exhibited extensive trans-regulatory activity influencing 429 and 161 genes, respectively. In particular, SNP 21:17035557 acted as a shared cis- and trans-regulatory hub, indicating hierarchical control of adipose gene networks. Functional enrichment analyses revealed significant involvement of cytoskeleton- and calcium-dependent pathways, highlighting the interplay between structural remodeling and metabolic regulation in adipose tissue. These findings provide a comprehensive, system-level view of adipose gene regulation in Hanwoo cattle and highlight candidate molecular targets for genome-assisted and precision breeding. Moreover, this study offers quantitative genomic resources that can support the development of prediction models and decision-support systems for improving carcass traits in Hanwoo breeding programs. Full article

(This article belongs to the Special Issue Transcriptomics in Livestock: Unveiling Genetic Insights for Enhanced Productivity and Health)

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16 pages, 1361 KB

Open AccessArticle

Development and Validation of an IMU Sensor-Based Behaviour-Alert Detection Collar for Assistance Dogs: A Proof-of-Concept Study

by Shelley Brady, Alan F. Smeaton, Hailin Song, Tomás Ward, Aoife Smeaton and Jennifer Dowler

Animals 2025, 15(21), 3081; https://doi.org/10.3390/ani15213081 - 23 Oct 2025

Abstract

Assistance dogs have shown promise in alerting to epileptic seizures in their owners, but current approaches often lack consistency, standardisation, and objective validation. This proof-of-concept study presents the development and initial validation of a wearable behaviour-alert detection collar developed for trained assistance dogs. [...] Read more.

Assistance dogs have shown promise in alerting to epileptic seizures in their owners, but current approaches often lack consistency, standardisation, and objective validation. This proof-of-concept study presents the development and initial validation of a wearable behaviour-alert detection collar developed for trained assistance dogs. It demonstrates the technical feasibility for automated detection of trained signalling behaviours. The collar integrates an inertial sensor and machine learning pipeline to detect a specific, trained alert behaviour of two rapid clockwise spins used by dogs to signal a seizure event. Data were collected from six trained dogs, resulting in 135 labelled spin alerts. Although the dataset size is limited compared to other machine learning applications, this reflects the real-world constraint that it is not practical for assistance dogs to perform excessive spin signalling during their training. Four supervised machine learning models (Random Forest, Logistic Regression, Naïve Bayes, and SVM) were evaluated on segmented accelerometer and gyroscope data. Random Forest achieved the highest performance (F1-score = 0.65; accuracy = 92%) under a Leave-One-DOG-Out (LODO) protocol. The system represents a novel step toward combining intentional canine behaviours with wearable technology, aligning with trends on the Internet of Medical Things. This proof-of-concept demonstrates technical feasibility and provides a foundation for future development of real-time seizure-alerting systems, representing an important first step toward scalable animal-assisted healthcare innovation. Full article

(This article belongs to the Special Issue Assistance Dogs: Health and Welfare in Animal-Assisted Services)

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22 pages, 2865 KB

Open AccessArticle

Taurine Supplementation in Low-Fishmeal of Golden Pompano (Trachinotus ovatus) Diets: Improving Intestinal Health and Alleviation of Inflammatory Response

by Zhanzhan Wang, Hongkai Ye, Zhong Huang, Jun Wang, Yun Wang, Wei Yu, Heizhao Lin, Zhenhua Ma and Chuanpeng Zhou

Animals 2025, 15(21), 3080; https://doi.org/10.3390/ani15213080 - 23 Oct 2025

Abstract

This research explored the effects of supplementing taurine in a low-fishmeal diet on the growth, hepatic antioxidant capacity, muscle quality, intestinal health, and alleviation of inflammatory response of golden pompano (Trachinotus ovatus). Over an eight-week period, 300 juvenile fish (initial weight [...] Read more.

This research explored the effects of supplementing taurine in a low-fishmeal diet on the growth, hepatic antioxidant capacity, muscle quality, intestinal health, and alleviation of inflammatory response of golden pompano (Trachinotus ovatus). Over an eight-week period, 300 juvenile fish (initial weight 9.4 ± 0.47 g) were randomly allocated into 12 net enclosures (1.0 × 1.0 × 1.5 m), with each treatment group comprising three replicate cages containing 25 specimens. The results demonstrated that an optimal taurine inclusion level of 1.0–1.5% significantly promoted growth, as evidenced by the increased weight gain rate (WGR) and specific growth rate (SGR). It also protected hepatic health by reducing alanine aminotransferase (ALT) activity and enhancing antioxidant capacity. Activation of the hepatic Nrf2/Keap-1/HO-1 signaling pathway increased the level of antioxidant gene expression, including catalase (CAT) and superoxide dismutase (SOD). In addition, the appropriate supplementation of taurine significantly down-regulated muscle hardness-related genes (cathepsin B (CatB) and cathepsin L (CatL)) and promoted the growth and differentiation of myoblasts, thus improving muscle quality. The chymotrypsin of fish fed the A25T10 diet was significantly higher than those in other groups (p < 0.05). The amylase (AMY) of fish fed the A25T15 diet was significantly higher than those in other groups (p < 0.05). The Chao1, Shannon, and Simpson of fish fed the A25T15 diet were significantly higher than those in other groups (p < 0.05). Proteobacteria were the most abundant in group A25T10. The relative abundance of Photobacterium rose in the A25RT10 group. In this study, taurine supplementation can down-regulate the expression of intestinal pro-inflammatory factors (interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8)) and up-regulate the expression of anti-inflammatory factor interleukin-10 (IL-10), enhance intestinal immunity, and improve intestinal digestion and absorption. Therefore, the addition of 1–1.5% taurine to low-fishmeal feeds can improve the growth performance of golden pompano. Full article

(This article belongs to the Special Issue Recent Advances in Nutritional Ingredients for Aquaculture)

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26 pages, 3746 KB

Open AccessFeature PaperArticle

Retrospective Analysis of Suspensory Ligament Branch Injuries in 70 Dressage Horses

by Ana Boado, Danica Pollard and Sue Dyson

Animals 2025, 15(21), 3079; https://doi.org/10.3390/ani15213079 - 23 Oct 2025

Abstract

There are no studies that have investigated factors influencing the outcome of dressage horses with suspensory ligament (SL) branch injuries. The aim was to determine if age, breed, work level, injury severity, anatomical localisation of injury, number of injured branches, periligamentous fibrosis, persistence [...] Read more.

There are no studies that have investigated factors influencing the outcome of dressage horses with suspensory ligament (SL) branch injuries. The aim was to determine if age, breed, work level, injury severity, anatomical localisation of injury, number of injured branches, periligamentous fibrosis, persistence of power Doppler signal or coexistent osteoarthritis of a metacarpophalangeal (MCP) or metatarsophalangeal (MTP) joint influenced the prognosis of 70 dressage horses. Outcome was defined as good (return to pre-injury level of work or higher), poor (return to a lower level of work) or retirement. Chi-squared or Fisher’s exact test and the Kruskal–Wallis test were used to identify relationships between variables of interest and follow-up outcome. Follow-up outcome was good in 44/70 horses (62.9%), poor in 13/70 (18.6.%) and 13/70 horses (19.1%) were retired due to no response to treatment. Ultrasonographic lesion grade (p = 0.07), cross-sectional area (CSA) of the SL (p = 0.96), CSA of the lesion (p = 0.28) and the lesion CSA as a percentage of the SL CSA (p = 0.40) were not associated with outcome. Power Doppler signal was present in 75.8% of injured branches at the initial examination. The severity of power Doppler signal was not associated with outcome (p = 0.20); however, persistence of power Doppler signal was negatively associated with outcome (p < 0.001). Other variables did not influence the follow-up outcome. Early recognition of SL branch injury is likely to result in a more favourable outcome with appropriate treatment and management. Full article

(This article belongs to the Section Equids)

25 pages, 2907 KB

Open AccessArticle

Machine-Milking Practices, Animal Welfare-Related Reactions and Quality of Milk Produced in Dairy Sheep Farms

by Dimitra V. Liagka, George C. Fthenakis, Stella N. Kalonaki, Konstantina S. Dimoveli, Daphne T. Lianou, Vasia S. Mavrogianni, Charalambia K. Michael, Mariangela Caroprese, Vassiliki Spyrou and Natalia G. C. Vasileiou

Animals 2025, 15(21), 3078; https://doi.org/10.3390/ani15213078 - 23 Oct 2025

Abstract

There is a scarcity of relevant information in the international literature regarding the welfare of dairy ewes during the milking process and the potential impact on the quality of the produced milk. Hence, there is a scope to further study potential interactions between [...] Read more.

There is a scarcity of relevant information in the international literature regarding the welfare of dairy ewes during the milking process and the potential impact on the quality of the produced milk. Hence, there is a scope to further study potential interactions between the milking process and animal welfare in dairy sheep flocks. The specific objectives of this study were (i) the evaluation of the frequency of reactions potentially associated with the reduced welfare of dairy sheep during the milking process, (ii) the identification of predictors associated with the presence of these reactions, and (iii) the evaluation of associations with the quality of milk produced on a farm. The study was conducted in 52 dairy sheep farms in Greece. The farms were visited, and the milking process was observed and monitored, and the reactions of ewes during the milking process were recorded; samples from the bulk-tank milk were collected for somatic cell and total bacterial counting. Univariable and multivariable analyses were performed. The median duration of milking per row was 7.9 min, and the median duration of the milking process was 105 min. The reaction observed less often from ewes was vocalisation (in 9.6% of farms), and the one observed more frequently was ‘spot stepping’ (in 98.1% of farms). In total, eight different predictors were identified for the displaying of the various reactions observed and recorded; of these, two were related to the milking parlour and six were related to the milking process. Moreover, three different predictors were identified for the milk quality parameters; of these, one was related to the milking parlour and two were related to the milking process. Finally, a mild correlation was seen between the proportion of ewes that attempted to remove the milking cluster, and the somatic cell counts in the bulk-tank milk (p = 0.023). The results confirmed that incorrect practices during machine-milking can be stress factors for dairy sheep. In light of the present findings, procedures by milkers should be carefully carried out, which will contribute to minimising the nervousness and aggravation of ewes during milking. Stress factors can adversely affect the quality of milk produced on the farm. The findings underline that maintenance of animal welfare is linked to high product quality. This connection can further extend to consumer perceptions: the ethical treatment of dairy sheep is considered as an important facet of the overall food quality and sustainable production. Full article

(This article belongs to the Special Issue Advances in Small Ruminant Welfare)

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17 pages, 4026 KB

Open AccessArticle

Integrated Whole-Transcriptome Analysis to Elucidate the Core Regulatory Network of circRNA Involved in Ovarian Development and Reproductive Capacity Differences in Sheep: circRNA2058-miR-9226-5p-MET Axis

by Bo Gu, Anqi Wang, Xinmiao Yu, Ying Li, Yao Cong and Huaizhi Jiang

Animals 2025, 15(21), 3077; https://doi.org/10.3390/ani15213077 - 23 Oct 2025

Abstract

(1) Background: This study aims to systematically identify key candidate genes and the regulatory networks governing ovarian development in sheep breeds with divergent fecundity. Focusing on elucidating the central regulatory roles of these factors during distinct ovarian developmental stages in highly prolific breeds, [...] Read more.

(1) Background: This study aims to systematically identify key candidate genes and the regulatory networks governing ovarian development in sheep breeds with divergent fecundity. Focusing on elucidating the central regulatory roles of these factors during distinct ovarian developmental stages in highly prolific breeds, the research seeks to reveal the mechanism by which multilevel regulatory networks synergistically determine ewe reproductive capacity. (2) Methods: This study utilized the ovaries from the low-fecundity sheep breed Ujumqin sheep, the high-fecundity breed small-tailed Han sheep, and various developmental stages of small-tailed Han sheep as research subjects. Through whole-transcriptome sequencing analysis, differentially expressed mRNAs(DEGs) and non-coding RNAs (ncRNAs) were screened, and a ceRNA regulatory network was constructed and subjected to bioinformatic analysis. The dual-luciferase reporter gene detection system was employed to validate the targeting relationships within the obtained key circRNA-miRNA-mRNA networks. Finally, qRT-PCR was used to verify the accuracy of the sequencing results. (3) Results: Our analysis constructed two distinct ceRNA networks: one from different fecundity groups (116 DECs, 46 DEMs, 82 DEGs) and another from different ovarian stages (186 DECs, 143 DEMs, 338 DEGs). Functional enrichment revealed key reproduction-related pathways, including Mitogen-Activated Protein Kinase(MAPK), Janus Kinase-Signal Transducer and Activator of Transcription(JAK-STAT), and WNT signaling in the fecundity comparison, and MAPK, Ras, WNT, Hippo signaling in the developmental stage comparison. Integrated analysis identified a core circRNA-miRNA-mRNA network, pinpointing circRNA2058-miR-9226-5p-MET as a central regulatory axis. The dual-luciferase assay confirmed that circRNA2058 acts as a sponge for miR-9226-5p, thereby mediating MET expression. qRT-PCR validation of randomly selected RNAs confirmed the sequencing reliability. (4) Conclusions: this study deciphers a synergistic regulatory network and identifies, for the first time, the pivotal circRNA2058-miR-9226-5p-MET ceRNA axis as an potential critical molecular switch driving follicular dominance in sheep. This discovery provides a molecular foundation for targeting core regulators of ovine reproductive efficiency and offers significant insights for innovative strategies in enhancing sheep reproduction. Full article

(This article belongs to the Section Small Ruminants)

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22 pages, 10841 KB

Open AccessArticle

Visceral Peritoneum Hyperpigmentation in Chickens Is Associated with DCT Expression

by Xu Dong, Qingchun Liu, Jiabao Xing, Guodong Mo, Muyang Li, Qinghua Nie, Jingting Shu and Zhenhui Li

Animals 2025, 15(21), 3076; https://doi.org/10.3390/ani15213076 (registering DOI) - 23 Oct 2025

Abstract

Hyperpigmentation of the visceral peritoneum (HVP) is a pigmentation abnormality in chickens that adversely affects carcass appearance, consumer acceptance, and poultry production. However, the genetic basis of HVP remains unclear. To investigate the causes and regulatory mechanisms of HVP, we employed high-performance liquid [...] Read more.

Hyperpigmentation of the visceral peritoneum (HVP) is a pigmentation abnormality in chickens that adversely affects carcass appearance, consumer acceptance, and poultry production. However, the genetic basis of HVP remains unclear. To investigate the causes and regulatory mechanisms of HVP, we employed high-performance liquid chromatography (HPLC), bulk RNA sequencing (RNA-seq), qRT-PCR, Western blotting, and siRNA interference. Additionally, single-cell RNA sequencing (scRNA-seq) was used to examine gene expression at the cellular level. Anatomical examination and hematoxylin and eosin (HE) staining revealed melanin deposition in the peritoneum of HVP-affected chickens. Spectrophotometric analysis at 500 nm showed significantly higher absorbance in the HVP group (p < 0.05), which correlated with the degree of pigmentation. HPLC confirmed the pigmentation as eumelanin, based on the pyrrole-2,3,5-tricarboxylic acid (PTCA) peak. RNA-seq identified 61 differentially expressed genes. Functional studies showed that dopachrome tautomerase (DCT) overexpression, combined with L-tyrosine (L-Tyr) supplementation, significantly increased melanin content (p < 0.05) and promoted melanocyte proliferation. In contrast, DCT silencing reduced melanin secretion and inhibited cell growth. ScRNA-seq analysis of over 9700 high-quality cells identified distinct melanocyte clusters, with DCT expression approximately 2.5-fold higher in melanocytes from the HVP group compared to the normal group. Furthermore, a DCT polymorphism (g.147917398 C > T) was identified as a potential marker for genetic selection (p-values = 0.033). These findings demonstrate that HVP is driven by DCT overexpression and excessive eumelanin deposition. DCT could serve as a molecular marker for genomic selection to improve poultry carcass quality and reduce economic losses in the poultry industry. Full article

(This article belongs to the Special Issue Genetic Diversity and Conservation of Local Poultry Breeds)

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25 pages, 3281 KB

Open AccessArticle

Chasing Pinna nobilis Survivors: Current Status in Spanish Open Coastal Waters

by Francesco Maresca, Elvira Álvarez, Lara Zafra, Iris E. Hendriks, Gaetano Catanese, Raul González, José Rafael García-March and Maite Vázquez-Luis

Animals 2025, 15(21), 3075; https://doi.org/10.3390/ani15213075 - 23 Oct 2025

Abstract

The largest and endemic bivalve of the Mediterranean Sea, Pinna nobilis, is on the brink of extinction after a mass mortality event (MME) that has affected its populations since autumn 2016. Since then, different actions have been performed to improve the conservation [...] Read more.

The largest and endemic bivalve of the Mediterranean Sea, Pinna nobilis, is on the brink of extinction after a mass mortality event (MME) that has affected its populations since autumn 2016. Since then, different actions have been performed to improve the conservation status of P. nobilis. The monitoring of survivors in open coastal systems along the Spanish Mediterranean coast showed, after an 8-year period since the start of the MME (2017–2024), that the geographical distribution of the survivors in open sea is currently concentrated in a few regions, with focal points of specimen density in Cap de Creus (Catalonia) and Menorca (Balearic Islands). During the exhaustive monitoring of individuals of P. nobilis, the active participation of citizen science became decisive, locating almost half of the survivors. Most individuals were found in marine protected areas, mainly in Posidonia oceanica meadows in the upper 15 m. As a safety measure, several survivors were translocated to safer areas, while evaluation of the impact of the translocation showed no demonstrable effects. The knowledge acquired during these years has highlighted the necessity for collaborative monitoring, specifically to understand the current critical situation of P. nobilis and to implement effective conservation measures for this emblematic species. Full article

(This article belongs to the Section Ecology and Conservation)

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17 pages, 556 KB

Open AccessFeature PaperArticle

Repeated Task Exposure and Sufficient Sleep May Mitigate ADHD-Related Cognitive Flexibility Impairments in Family Dogs

by Tímea Kovács, Vivien Reicher, Barbara Csibra, Melitta Csepregi, Kíra Kristóf and Márta Gácsi

Animals 2025, 15(21), 3074; https://doi.org/10.3390/ani15213074 (registering DOI) - 23 Oct 2025

Abstract

The family dog is a valid model for studying complex human functions and psychological disorders such as Attention-Deficit/Hyperactivity Disorder (ADHD). Based on prior human research indicating impairments in cognitive flexibility related to ADHD, this study investigates the association between dogs’ ADHD-like traits and [...] Read more.

The family dog is a valid model for studying complex human functions and psychological disorders such as Attention-Deficit/Hyperactivity Disorder (ADHD). Based on prior human research indicating impairments in cognitive flexibility related to ADHD, this study investigates the association between dogs’ ADHD-like traits and reversal learning performance. Since sleep improves learning both in humans and dogs, we also examined its impact in this context. Family dogs (N = 64) completed a two-way choice spatial reversal learning task, followed by a one-hour non-invasive sleep electroencephalography (EEG), and then a second reversal task. We used a validated human analogue questionnaire to assess ADHD. Dogs with higher ADHD scores required more trials to pass the first reversal test, but not after sleep. Electrode application was slower and sleep measurement more likely to fail in dogs with higher ADHD scores. Performance improved more from pre- to post-sleep in high-ADHD dogs if they spent at least 40% of the recording asleep. Our findings align with the human literature showing associations between ADHD and cognitive flexibility in dogs. The main novelty here is the ADHD-related potential benefits of repeated task exposure after sufficient sleep on cognitive flexibility. Cognitive training offers a promising direction to mitigate ADHD-related impairments in dogs. Full article

(This article belongs to the Section Human-Animal Interactions, Animal Behaviour and Emotion)

14 pages, 1238 KB

Open AccessArticle

Fermented Chinese Herbs Improved Egg Production, Egg Shell Quality, and Egg Yolk Cholesterol of Laying Hens by Regulating Estrogen, Lipid Metabolism, and Calcium Metabolism

by Xinyu Liu, Yue He, Yuhan Cao, Xin Wang, Ye Yang and Jiao Song

Animals 2025, 15(21), 3073; https://doi.org/10.3390/ani15213073 - 23 Oct 2025

Abstract

The present study investigated the effects of fermented Chinese herbal (FCH) compounds on the egg production, egg shell quality, and egg yolk cholesterol of laying hens. A total of 1260 Hy-Line pink laying hens, 34 weeks old, were randomly divided into three groups, [...] Read more.

The present study investigated the effects of fermented Chinese herbal (FCH) compounds on the egg production, egg shell quality, and egg yolk cholesterol of laying hens. A total of 1260 Hy-Line pink laying hens, 34 weeks old, were randomly divided into three groups, with six replicates per group and 70 hens per replicate, as follows: the control group (CON group) was fed a diet without FCH compounds, and the 2% FCH group and the 3% FCH group were fed a diet supplemented with 2% FCH and 3% FCH, respectively. The results show that the FCH compound significantly increased the laying rate compared to the CON group (p < 0.05). Analyses of the serum biochemical indices showed that supplementation with FCH compound significantly decreased the levels of total cholesterol (TC), total triglyceride (TG), high-density lipoprotein cholesterol (HLDL-c), low-density lipoprotein cholesterol (LDL-c), very-low-density lipoprotein cholesterol (VLVL-c), aspartate transaminase (AST), and alanine aminotransferase (ALT) (p < 0.05) and increased the serum total bile acids, follicle-stimulating hormone (FSH), luteinizing hormone (LH), and 17-β-Estradiol (E2) levels (p < 0.05). The FCH group significantly increased the activity of superoxide dismutase (SOD) and total antioxidant capacity and decreased malondialdehyde (MDA) levels in the liver and uterus compared to the CON (p < 0.05). FCH supplementation was also associated with improved egg quality, seen through factors including enhanced yolk color, albumen height, Haugh unit score, eggshell strength, and thickness and reduced egg breaking rate and TC and TG contents in egg yolk. The gene expression analyses showed that FCH supplementation significantly increased the calcium metabolism-related gene expression (CaBP-D_28k, NCX, VDR, CYP₂₇B₁, OPN, PMCA, CA₂) in duodenum, kidney, and uterus tissues compared to the CON group (p < 0.05). FCH significantly repressed FAS and HMGCR mRNA expression and enhanced CYP7A1 mRNA expression in the liver (p < 0.05). These results indicate that diet supplementation with FCH compounds may improve egg quality by regulating reproductive hormones, lipid metabolism, and calcium metabolism. Full article

(This article belongs to the Section Animal Nutrition)

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Open AccessArticle

In Vivo Ultrasonographic Assessment of Bone Mineral Density and Its Impact on Semen Quality in Boars

by Miaomiao Liao, Xinyu Liu, Hengxi Wei, Li Li and Shouquan Zhang

Animals 2025, 15(21), 3072; https://doi.org/10.3390/ani15213072 - 23 Oct 2025

Abstract

Bone mineral density (BMD) is a key indicator of skeletal health in boars that influences their reproductive performance. Systematic research on the relationship between BMD and semen quality in adult boars of different breeds and ages is scarce. This study used quantitative ultrasound [...] Read more.

Bone mineral density (BMD) is a key indicator of skeletal health in boars that influences their reproductive performance. Systematic research on the relationship between BMD and semen quality in adult boars of different breeds and ages is scarce. This study used quantitative ultrasound (QUS) technology to measure BMD in 492 adult and 208 replacement boars. The boars were divided into four equal groups based on descending BMD rankings to analyze correlations with semen quality. Simultaneously, a 25-hydroxyvitamin D₃ (25-OH-D₃) intervention trial was conducted on 150 adult Duroc boars. A control group and four dose groups (50 μg, 125 μg, 200 μg, and 250 μg) were established. After 90 days, the boars’ semen quality, reproductive hormone levels, and bone metabolism indicators were evaluated. The results showed no significant differences in BMD between adult and replacement boars. However, adult Landrace exhibited significantly higher BMD than Duroc and Yorkshire (p < 0.01). Within the BMD groups, Group D boars had significantly higher rates of sperm abnormality than Groups A and B (p < 0.01), and this trend was consistent across breeds. The 25-OH-D₃ intervention results indicated that the 250 μg dosage produced the optimal effect. In this group, boar semen motility significantly improved while sperm abnormality rates significantly decreased. Concurrently, levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), serum osteocalcin (OC), and BMD all increased to some degree. In summary, boar BMD is significantly influenced by breed and age and is closely correlated with the rate of sperm abnormality rate. Supplementing with 250 μg of 25-OH-D₃ effectively enhances reproductive hormone secretion, improves semen quality, and promotes bone formation. This demonstrates its potential value in breeding and nutritional regulation applications. Full article

(This article belongs to the Section Pigs)

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Animals, Volume 15, Issue 21 (November-1 2025) – 41 articles

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