Animals

24 pages, 341 KB

Open AccessArticle

Impact of Living Environment on Attachment Behaviour in Domestic Cats from Private Homes and Shelters

by Isabelle Kappel, Bianca Materne and Udo Gansloßer

Animals 2025, 15(24), 3521; https://doi.org/10.3390/ani15243521 - 5 Dec 2025

Since cats often struggle to acclimate to laboratory settings, a key requirement for effective testing, we adapted Ainsworth’s Strange Situation Test (SST) for use in environments familiar to the cats. We examined 82 cat–caregiver dyads from private households and animal shelters using a [...] Read more.

Since cats often struggle to acclimate to laboratory settings, a key requirement for effective testing, we adapted Ainsworth’s Strange Situation Test (SST) for use in environments familiar to the cats. We examined 82 cat–caregiver dyads from private households and animal shelters using a standardised five-phase video procedure comprising distinct phases of presence, separation, and reunion between the cats and their human caregivers. Seven attachment-related behaviours were recorded via focal animal sampling and analysed using mixed models. Significant differences were observed in exploratory behaviour, physical contact, play (social, object, and locomotor), passive behaviour, and vocalisation. Cats from private homes exhibited more exploratory and play behaviour, particularly after reuniting with their caregiver, indicating a positive influence of the familiar person’s presence. Cats from animal shelters exhibited reduced play behaviour, more passive behaviour, and more frequent purring, particularly following reunion. Physical contact increased following separations, indicating attachment. Meowing was most frequent during absences and reunions, possibly reflecting stress and renewed social contact. Outdoor access emerged as a potential influencing factor for exploratory behaviour. In summary, attachment behaviours in cats were significantly influenced by the living environment, access to the outdoors, and the presence of familiar or unfamiliar individuals. Certain test phases elicited particularly strong behavioural responses. Full article

(This article belongs to the Special Issue Second Edition: Research on the Human–Companion Animal Relationship)

11 pages, 902 KB

Open AccessArticle

Mass Spectrometry-Based Proteomic Analysis of Porcine Reproductive and Respiratory Syndrome Virus NSP9 Protein with Host Proteins

by Wei Wen, Yuhang Liu, Wenqiang Wang, Zhenbang Zhu and Xiangdong Li

Animals 2025, 15(24), 3520; https://doi.org/10.3390/ani15243520 - 5 Dec 2025

Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) nonstructural protein 9 (NSP9), the viral RNA-dependent RNA polymerase (RdRp), is essential for viral replication but its comprehensive host interactome remains uncharacterized. This study employed co-immunoprecipitation coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) to systematically identify [...] Read more.

Porcine reproductive and respiratory syndrome virus (PRRSV) nonstructural protein 9 (NSP9), the viral RNA-dependent RNA polymerase (RdRp), is essential for viral replication but its comprehensive host interactome remains uncharacterized. This study employed co-immunoprecipitation coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) to systematically identify NSP9-associated host proteins. We identified 222 high-confidence host interactors, with Gene Ontology and KEGG pathway analyses revealing significant enrichment in RNA/DNA-binding proteins, ubiquitin-proteasome pathways, metabolic regulators (amino acid/lipid biosynthesis), endoplasmic reticulum processing, and cell cycle components. Protein-protein interaction network analysis further delineated six functional modules involved in RNA processing, vesicular transport, and innate immunity. Crucially, validation studies confirmed direct binding between NSP9 and key candidates (CAPZ1, PSMA3, CDK1, USP48). Functional assessment demonstrated that CDK1 overexpression significantly inhibited PRRSV replication, implicating CDK1 as a host restriction factor. These findings collectively unveil the multifaceted role of NSP9 in subverting host machinery while identifying novel host defense mechanisms and potential targets for antiviral development against PRRSV. Full article

(This article belongs to the Section Pigs)

12 pages, 693 KB

Open AccessArticle

Fluorescence-Guided Thoracoscopic Surgery Using Indocyanine Green (ICG) in Canine Cadavers: A Descriptive Evaluation of Video-Assisted (VATS) and Robot-Assisted (RATS) Approaches

by Francisco M. Sánchez-Margallo, Lucía Salazar-Carrasco, Manuel J. Pérez-Salazar and Juan A. Sánchez-Margallo

Animals 2025, 15(24), 3519; https://doi.org/10.3390/ani15243519 - 5 Dec 2025

Abstract

Precise intraoperative identification of the canine thoracic duct remains challenging due to anatomical variability and limited visualization. This exploratory cadaveric feasibility study aimed to describe the technical applicability of fluorescence-guided thoracic duct mapping using video-assisted thoracoscopy (VATS) and robot-assisted thoracoscopy (Versius™ system). Four [...] Read more.

Precise intraoperative identification of the canine thoracic duct remains challenging due to anatomical variability and limited visualization. This exploratory cadaveric feasibility study aimed to describe the technical applicability of fluorescence-guided thoracic duct mapping using video-assisted thoracoscopy (VATS) and robot-assisted thoracoscopy (Versius™ system). Four adult Beagle cadavers underwent bilateral thoracoscopic exploration after intranodal injection of indocyanine green (ICG, Verdye^®, 0.05 mg/kg; 0.5 mL). Near-infrared (NIR) fluorescence imaging enabled real-time visualization of the thoracic duct and its branches. Fluorescence quality was quantitatively characterized using signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), and contrast resolution (CR) calculated from standardized image frames. Both approaches achieved successful duct identification in all cadavers. VATS provided brighter overall fluorescence, whereas the robotic-assisted approach offered stable imaging, enhanced instrument dexterity, and improved duct-to-background discrimination. These findings confirm the feasibility of fluorescence-guided thoracic duct identification using both minimally invasive modalities in canine cadavers. The standardized assessment of optical parameters proposed here may support future in vivo studies to optimize imaging protocols and evaluate the clinical impact of fluorescence-guided thoracic duct surgery in dogs. Full article

(This article belongs to the Section Companion Animals)

19 pages, 1495 KB

Open AccessArticle

Integrative Genomics and Multi-Tissue Transcriptomics Identify Key Loci and Pathways for Hypoxia Tolerance in Grass Carp

by Wenwen Wang, Mengyang Chang, Suxu Tan, Yiming Hu, Xinlu Ren, Hongtao Xue, Lizheng Gao, Xiao Cao, Ya Wang, Qiyu Li and Zhenxia Sha

Animals 2025, 15(24), 3518; https://doi.org/10.3390/ani15243518 - 5 Dec 2025

Abstract

Hypoxia is a critical environmental stressor in aquaculture, significantly affecting the survival and growth performance of cultured fish. To explore the genetic basis of hypoxia tolerance in grass carp (Ctenopharyngodon idella), we integrated genome-wide association analysis (GWAS) and multi-tissue transcriptome profiling. [...] Read more.

Hypoxia is a critical environmental stressor in aquaculture, significantly affecting the survival and growth performance of cultured fish. To explore the genetic basis of hypoxia tolerance in grass carp (Ctenopharyngodon idella), we integrated genome-wide association analysis (GWAS) and multi-tissue transcriptome profiling. A total of 2000 grass carp were subjected to hypoxic stress, from which the 150 most hypoxia-intolerant (HI) and 150 most hypoxia-tolerant (HT) individuals were selected based on the time to loss of equilibrium (LOE). GWAS using 3,730,919 SNPs and 851,595 InDels identified 21 SNPs and 6 InDels associated with hypoxia tolerance. Two SNPs on chromosomes 10 and 13 reached genome-wide significance, accounting for 2.7% and 4.8% of the phenotypic variance explained (PVE), respectively. Validation of identified SNPs was performed using kompetitive allele-specific PCR (KASP) analysis. Candidate genes within ±50 kb of these variants were enriched in steroid biosynthesis, insulin signaling, and glycosphingolipid biosynthesis pathways. Transcriptomic analysis of six tissues (brain, gill, intestine, kidney, liver, and spleen) revealed 1620, 1221, 796, 246, 210, and 58 differentially expressed genes (DEGs) in the HT group compared to the HI group, respectively. DEGs in the brain were primarily enriched in steroid metabolic processes and angiogenesis regulation, while those in kidney and spleen DEGs were associated with oxygen transport and erythrocyte development. Integrated analysis of GWAS and transcriptome data identified 16 shared genes, including usf1 and trpv4. These findings reveal key genomic loci and molecular pathways underlying hypoxia tolerance in grass carp, providing valuable markers for future selective breeding programs. Full article

(This article belongs to the Special Issue Advances in Research on Functional Genes and Economic Traits in Fish)

22 pages, 2269 KB

Open AccessArticle

Dietary Scutellaria baicalensis and Lonicera japonica Extract Supplementation Attenuates Oxidative Stress and Improves Reproductive Performance in Sows

by Nuan Wang, Huiyuan Lv, Wei Chai, Hanting Ding, Junjie Yang, Hanyu Jing, Fang Chen and Wutai Guan

Animals 2025, 15(24), 3517; https://doi.org/10.3390/ani15243517 - 5 Dec 2025

Abstract

Harnessing the powerful antioxidant and anti-inflammatory properties of Scutellaria baicalensis and Lonicera japonica (SL), SL extract emerges as a natural and effective dietary strategy to enhance sow reproductive performance and overall health. In this study, 100 multiparous Duroc × Landrace × Yorkshire sows [...] Read more.

Harnessing the powerful antioxidant and anti-inflammatory properties of Scutellaria baicalensis and Lonicera japonica (SL), SL extract emerges as a natural and effective dietary strategy to enhance sow reproductive performance and overall health. In this study, 100 multiparous Duroc × Landrace × Yorkshire sows were assigned to either a control diet or a diet supplemented with 0.05% SL extract (n = 100), beginning on day 85 of gestation and continuing until day 21 of lactation, with 50 sows in each group. Duroc boars were the source of semen for artificial insemination. While SL supplementation did not affect litter size, birth weight, or milk composition, it significantly reduced piglet mortality during lactation, from 13.11% to 9.72% (p < 0.05). Compared with the control group, feed intake of sows in the SL group increased from 4.56 kg to 4.70 kg (p < 0.01) during lactation. Furthermore, SL extract enhanced the antioxidant capacity of the sows, reduced malondialdehyde and levels of IL-1β, IL-6, and TNF-α, and increased the plasma soluble cluster of differentiation 14 (sCD14) concentrations (p < 0.05). In vitro, pretreatment of mammary epithelial cells with SL extract (2 μg/mL for 24 h) before lipopolysaccharide stimulation significantly upregulated antioxidant markers, suppressed pro-inflammatory cytokine mRNA expression, and inhibited activation of the NF-κB and MAPK pathways (p < 0.05). These findings highlight the potential of SL extract as a natural feed additive to mitigate oxidative stress and inflammation, ultimately supporting improved reproductive performance and health in sows. Full article

(This article belongs to the Special Issue Maternal Nutrition and Neonatal Development of Pigs)

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17 pages, 2584 KB

Open AccessArticle

CKMT2 Promotes Breast Muscle Growth in Qiangying Ducks via Enhancing Myoblast Proliferation and Differentiation

by Longfei Xie, Dongsheng Wu, Wanli Yang, Ya Li, Jie Zhang and Zhaoyu Geng

Animals 2025, 15(24), 3516; https://doi.org/10.3390/ani15243516 - 5 Dec 2025

Abstract

The duck meat industry is economically vital in animal husbandry; however, the genetic mechanisms governing skeletal muscle development and muscle yield remain incompletely understood. In this study, RNA sequencing (RNA-seq) was used to identify differentially expressed genes (DEGs) in breast muscle tissues between [...] Read more.

The duck meat industry is economically vital in animal husbandry; however, the genetic mechanisms governing skeletal muscle development and muscle yield remain incompletely understood. In this study, RNA sequencing (RNA-seq) was used to identify differentially expressed genes (DEGs) in breast muscle tissues between high and low breast muscle weight (BMW) groups of Qiangying ducks. A candidate gene, CKMT2 (mitochondrial creatine kinase 2), was subsequently validated at the cellular and individual levels. RNA-seq analysis identified 540 DEGs, including 411 upregulated and 129 downregulated genes. GO and KEGG enrichment analyses revealed enrichment in fatty acid metabolism, fibrinolysis, and PPAR signaling pathways—processes closely linked to energy homeostasis and muscle growth. Functional validation demonstrated that CKMT2 overexpression significantly promoted myoblast proliferation and myotube differentiation (p < 0.05), while knockdown exerted the opposite effects. At the genetic level, the GG genotype of CKMT2 SNP (G.76,602,082 G>A) was associated with significantly higher BMW compared to the GA and AA genotypes (p < 0.05). These findings provide novel insights into the molecular basis of muscle weight variation in meat ducks and establish CKMT2 as a key regulator of skeletal muscle growth via modulation of myoblast proliferation and differentiation. This study contributes to the theoretical foundation for improving duck muscle yield through genetic selection. Full article

(This article belongs to the Section Poultry)

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17 pages, 3606 KB

Open AccessArticle

Dietary Fagopyrum dibotrys Extract Supplementation: Impacts on Growth Performance, Immune Response, Intestinal Morphology, and Microbial Community in Broiler Chickens Infected with Escherichia coli O157

by Jiang Chen, Gaoxiang Ai, Pingwen Xiong, Wenjing Song, Guohua Liu, Qipeng Wei, Xiaolian Chen, Zhiheng Zou and Qiongli Song

Animals 2025, 15(24), 3515; https://doi.org/10.3390/ani15243515 - 5 Dec 2025

Abstract

This study explored the efficacy of dietary Fagopyrum dibotrys extract (FDE) in mitigating Escherichia coli O157 (E. coli) infections in broilers. A total of 240 one-day-old male Shengze 901 broilers were randomly allocated to four groups (with 10 broilers per group): [...] Read more.

This study explored the efficacy of dietary Fagopyrum dibotrys extract (FDE) in mitigating Escherichia coli O157 (E. coli) infections in broilers. A total of 240 one-day-old male Shengze 901 broilers were randomly allocated to four groups (with 10 broilers per group): CON (basal diet), COLI (basal diet + E. coli challenge), FDE (basal diet + 500 mg/kg FDE), and FDEC (basal diet + 500 mg/kg FDE + E. coli challenge). The results showed that E. coli challenge reduced the average daily gain (ADG) and average daily feed intake (ADFI), increased the feed conversion ratio (FCR) and cecal E. coli load, impaired the intestinal mucosa, and induced intestinal inflammatory responses (p < 0.05). FDE supplementation improved growth performance, increased duodenal villus height and villus/crypt ratio; reduced serum interleukin (IL)-1β, tumor necrosis factor-α (TNF-α), diamine oxidase (DAO), and endotoxin levels; and lowered cecal E. coli counts (p < 0.05). Molecularly, FDE supplementation upregulated Occludin, Claudin-1, and ZO-1 gene expressions, and downregulated jejunal TLR4 and MyD88 mRNA levels. Microbiome analysis revealed that FDE increased the relative abundance of Faecalibacterium and alleviated the E. coli-induced reduction in Clostridia_UCG-014. In conclusion, dietary supplementation with 500 mg/kg FDE could mitigate colibacillosis-related intestinal damage and inflammatory responses. Full article

(This article belongs to the Section Poultry)

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18 pages, 1782 KB

Open AccessArticle

Dietary Net Energy Concentration Affects Growth Performance, Carcass Traits, Intramuscular Fatty Acid Profile, and Cecal Microbiota of Pigs with Restricted Feed Allowance

by Qinfeng Hu, Wanxin Xiang, Youzhi Pu, Yong Zhang, Pan Zhou, Tiande Zou, Zhengjun Xie, Zhiqing Wu, Xiang Ao, Jinming You and Honglin Yan

Animals 2025, 15(24), 3514; https://doi.org/10.3390/ani15243514 - 5 Dec 2025

Abstract

This study investigated the effects of different dietary net energy (NE) concentrations on the growth performance, carcass traits, meat quality, and cecal microbiota of feed restricted pigs. In total, 32 Sichuan-Tibetan black pigs with similar initial body weights (25.98 ± 0.27 kg) were [...] Read more.

This study investigated the effects of different dietary net energy (NE) concentrations on the growth performance, carcass traits, meat quality, and cecal microbiota of feed restricted pigs. In total, 32 Sichuan-Tibetan black pigs with similar initial body weights (25.98 ± 0.27 kg) were divided into four groups: a control group (ad libitum feeding, 2330 kcal NE kg⁻¹) and three treatment groups (twice-daily feeding with NE concentrations of 2330, 2370, and 2410 kcal kg⁻¹, respectively). This feeding trial lasted for 19 weeks. Decreasing feeding frequency reduced the final body weight, average daily gain (ADG), average daily feed intake (ADFI), and backfat thickness (p < 0.05). Elevation of NE concentration increased the final body weight, ADG, and fat deposition (p < 0.05), which eventually led to an improvement in feed efficiency (p < 0.05). The expression of lipid-metabolism-related genes was significantly downregulated as NE concentration increased under a restricted feeding regime (p < 0.05). Cecal microbiota analysis showed that a high NE concentration decreased microbial alpha-diversity (p < 0.05). These findings suggest that under the restricted feeding condition, 2370 kcal NE kg⁻¹ can improve feed efficiency and optimize carcass composition simultaneously, which are associated with the changes in gut microbiota structure and the lipid-metabolism-related gene expression. Full article

(This article belongs to the Section Pigs)

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14 pages, 5676 KB

Open AccessArticle

A20 Attenuates Inflammatory Injury in Bovine Endometrial Epithelial Cells Through Autophagy-Mediated NLRP3 Inflammasome Inactivation

by Yongshuai Jiang, Fan Fei, Xiaoyu Wang, Yeqi Jiang, Kangjun Liu, Long Guo, Luying Cui, Heng Wang, Junsheng Dong and Jianji Li

Animals 2025, 15(24), 3513; https://doi.org/10.3390/ani15243513 - 5 Dec 2025

Abstract

Endometritis, an inflammatory disease of the uterine endometrial tissue, is a major reproductive disorder in dairy cattle that causes extensive damage to endometrial epithelial cells. Excessive activation of the NLRP3 inflammasome is strongly associated with inflammatory pathology. Autophagy plays a critical role in [...] Read more.

Endometritis, an inflammatory disease of the uterine endometrial tissue, is a major reproductive disorder in dairy cattle that causes extensive damage to endometrial epithelial cells. Excessive activation of the NLRP3 inflammasome is strongly associated with inflammatory pathology. Autophagy plays a critical role in clearing damaged proteins, organelles, and intracellular pathogens. Additionally, the zinc finger protein A20 exhibits potent anti-inflammatory effects across various inflammatory conditions. However, the roles of A20 and autophagy in regulating the NLRP3 inflammasome in BEECs remain poorly defined. This study shows that LPS significantly increased IL-1β expression, Caspase-1 activity, and lactate dehydrogenase (LDH) levels, while inducing numerous vesicular protrusions and membrane pores, resulting in severe inflammatory injury. A20 overexpression mitigated LPS-induced NLRP3 inflammasome activation and alleviated inflammatory injury. Conversely, autophagy inhibition or A20 silencing intensified LPS-induced NLRP3 inflammasome activation and inflammatory injury. Further analysis revealed that A20 promotes autophagy, and its inhibitory effect on the NLRP3 inflammasome was diminished when autophagy was suppressed. In conclusion, A20 reduces LPS-induced inflammatory injury in BEECs by enhancing autophagy and suppressing NLRP3 inflammasome activation. These results uncover a novel regulatory role for A20 in controlling excessive NLRP3 inflammasome activation in BEECs, suggesting its potential as a therapeutic target for bovine endometritis. Full article

(This article belongs to the Special Issue New Insights into Exploring the Development of the Animal Uterus during Pregnancy)

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15 pages, 5739 KB

Open AccessArticle

Distinct Effects of GnRH Immunocastration Versus Surgical Castration on Gut Microbiota

by Fanli Kong, Ruohan Yang, Xingyu Zhou, Yuanyuan Shen, Wenhao Wei, Xianyin Zeng, Xiaogang Du and Xinfa Han

Animals 2025, 15(24), 3512; https://doi.org/10.3390/ani15243512 - 5 Dec 2025

Abstract

Surgical castration, a common practice in animal husbandry, raises animal welfare concerns and adversely affects growth performance. Active immunization against gonadotropin-releasing hormone (GnRH) provides a non-surgical alternative. Both methods ultimately suppress sex hormone production, but their comparative effects on the gut microbiota, a [...] Read more.

Surgical castration, a common practice in animal husbandry, raises animal welfare concerns and adversely affects growth performance. Active immunization against gonadotropin-releasing hormone (GnRH) provides a non-surgical alternative. Both methods ultimately suppress sex hormone production, but their comparative effects on the gut microbiota, a crucial regulator of host health and metabolism, remain unclear. Here, 60 Sprague Dawley (SD) rats were randomly allocated into three groups—control (n = 20; 10 female and 10 male), surgical castration (n = 20; 10 female and 10 male), and GnRH immunocastration groups (n = 20; 10 female and 10 male)—at 4–5 weeks of age to comparatively investigate the impacts of surgical versus GnRH immunocastration on the gut microbiota. Our study demonstrated GnRH immunocastration significantly reduced gonadal weight, effectively suppressing gonadal development to a level comparable to surgical castration. However, the two methods induced distinct, sex-dependent shifts in the gut microbiota. Surgical castration reduced the gut microbial community diversity, whereas the community structure of GnRH immunocastrated rats more closely resembled that of the control group, indicating a milder impact on the microbial diversity and composition. Notably, GnRH immunocastration resulted in higher microbial alpha diversity than surgical castration in both female and male SD rats. Specific bacterial genera, such as Clostridia_UCG014, Lactobacillus, and Lachnospiraceae_UCG006, were similarly altered in both surgical castration and GnRH immunocastration female SD rats, while Intestinimonas and Erysipelatoclostridiaceae_UCG004 were concurrently changed in male SD rats. Conversely, Eubacterium_nodatum_group exhibited opposite responses, increasing with GnRH immunocastration but decreasing with surgical castration in male SD rats. Functional prediction revealed fundamental sex differences in microbial metabolic pathways. In females, nitrogen metabolism, glyoxylate/dicarboxylate metabolism, and mismatch repair were changed, while the pathways involved in siderophore biosynthesis, the citrate cycle (TCA cycle), genetic information processing, and amino acid metabolism were changed in male SD rats. In conclusion, GnRH immunocastration appears to be a less disruptive intervention, better preserving microbial diversity and inducing a unique functional profile. These findings highlighted the importance of considering the castration method’s impact on the gut microbial ecosystem in animal production and provided insights for developing humane and effective approaches to animal population control. Full article

(This article belongs to the Section Animal Physiology)

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18 pages, 2606 KB

Open AccessArticle

Divergent Embryo Responses to Chemical Cues in Two Freshwater Fishes with Different Parental Care Strategies

by Ning Zhang, Qinlei Li, Minghui Li, Chris K. Elvidge, Chuke Deng, Deshou Wang, Shijian Fu and Jigang Xia

Animals 2025, 15(24), 3511; https://doi.org/10.3390/ani15243511 - 5 Dec 2025

Abstract

Chemical information is one of the most important modes of communication among aquatic organisms. However, it remains unclear whether fish embryos are innately responsive to ecologically-related chemical cues from the cues of their cohort competitors, potential caring or risky cues from parents, or [...] Read more.

Chemical information is one of the most important modes of communication among aquatic organisms. However, it remains unclear whether fish embryos are innately responsive to ecologically-related chemical cues from the cues of their cohort competitors, potential caring or risky cues from parents, or chemical alarm cues from damaged embryos, and whether these response patterns differ between species that model alternative parental care strategies. Here, we examined embryo responses in heart rate and incubation performance to different conspecific chemical cues (companion embryo odours, maternal odours, maternal + companion odours, embryonic alarm cues, or water controls) in zebrafish (Danio rerio; no parental care) and Nile tilapia (Oreochromis niloticus; high parental care). Zebrafish embryos did not respond to companion embryo odours but did respond similarly to maternal odours and alarm cues consistent with detecting elevated risk levels. In contrast, Nile tilapia embryos only demonstrated significant risk responses to embryonic alarm cues. These findings indicate that embryos of both species can innately recognize alarm cues but differ in their response patterns to conspecific odours. The results highlight that parental care strategies may influence intergenerational chemical communication in fish embryos. Full article

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

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14 pages, 2730 KB

Open AccessArticle

Breeding and Disease Resistance Evaluation of a New Largemouth Bass (Micropterus salmoides) Strain Resistant to Largemouth Bass Virus (LMBV)

by Wenxian Li, Pinhong Li, Xia Luo, Ningqiu Li, Qiang Lin, Hongru Liang, Yinjie Niu, Baofu Ma, Wenwen Xiao and Xiaozhe Fu

Animals 2025, 15(24), 3510; https://doi.org/10.3390/ani15243510 - 5 Dec 2025

Abstract

Largemouth bass virus (LMBV) has caused severe impacts on the largemouth bass aquaculture industry in China. Breeding new virus-resistant strains is a fundamental strategy to address the LMBV challenge. In this study, a candidate LMBV-resistant population of largemouth bass was developed using the [...] Read more.

Largemouth bass virus (LMBV) has caused severe impacts on the largemouth bass aquaculture industry in China. Breeding new virus-resistant strains is a fundamental strategy to address the LMBV challenge. In this study, a candidate LMBV-resistant population of largemouth bass was developed using the “Youlu No. 3” as the base population. Through three consecutive generations of selective breeding, a new LMBV-resistant strain was successfully established. To evaluate the LMBV resistance of different breeding generations, four key indicators, including survival rate post-LMBV infection, LMBV viral load, the expression of immune-related genes (GADD45b, FOXO3, TNF-α, IFN-γ, and IL-10), and antioxidant enzyme activities (GSH and AKP), were analyzed in this study. Notably, the F3 generation exhibited significantly lower viral loads in liver tissues after LMBV infection compared to the F1, F2, and control groups. Furthermore, the F3 generation showed a markedly higher survival rate at 14 days post-challenge, indicating enhanced disease resistance. Additionally, both the F2 and F3 generations had increased expression levels of the immune-related genes and elevated activities of antioxidant-related enzymes. These results collectively indicate that the F3 generation possesses stronger LMBV resistance than the F2, F1, and control groups. This study provides effective strategy for addressing LMBV disease in largemouth bass at the source and is of great significance for promoting the healthy and sustainable development of the largemouth bass aquaculture industry. Full article

(This article belongs to the Section Aquatic Animals)

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24 pages, 3036 KB

Open AccessArticle

MPG-SwinUMamba: High-Precision Segmentation and Automated Measurement of Eye Muscle Area in Live Sheep Based on Deep Learning

by Zhou Zhang, Yaojing Yue, Fuzhong Li, Leifeng Guo and Svitlana Pavlova

Animals 2025, 15(24), 3509; https://doi.org/10.3390/ani15243509 - 5 Dec 2025

Abstract

Accurate EMA assessment in live sheep is crucial for genetic breeding and production management within the meat sheep industry. However, the segmentation accuracy and reliability of existing automated methods are limited by challenges inherent to B-mode ultrasound images, such as low contrast and [...] Read more.

Accurate EMA assessment in live sheep is crucial for genetic breeding and production management within the meat sheep industry. However, the segmentation accuracy and reliability of existing automated methods are limited by challenges inherent to B-mode ultrasound images, such as low contrast and noise interference. To address these challenges, we present MPG-SwinUMamba, a novel deep learning-based segmentation network. This model uniquely combines the state-space model with a U-Net architecture. It also integrates an edge-enhancement multi-scale attention module (MSEE) and a pyramid attention refinement module (PARM) to improve the detection of indistinct boundaries and better capture global context. The global context aggregation decoder (GCAD) is employed to precisely reconstruct the segmentation mask, enabling automated measurement of the EMA. Compared to 12 other leading segmentation models, MPG-SwinUMamba achieved superior performance, with an intersection-over-union of 91.62% and a Dice similarity coefficient of 95.54%. Additionally, automated measurements show excellent agreement with expert manual assessments (correlation coefficient r = 0.9637), with a mean absolute percentage error of only 4.05%. This method offers non-invasive and efficient and objective evaluation of carcass performance in live sheep, with the potential to reduce measurement costs and enhance breeding efficiency. Full article

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

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15 pages, 298 KB

Open AccessArticle

Farmer and Farm-Level Predictors of Pain Perception and Management During Routine Pig Husbandry Practices in Eastern Cape, South Africa

by Asemahle Tyutwana, Ziyanda Mpetile, Olusegun Oyebade Ikusika and Oluwakamisi Festus Akinmoladun

Animals 2025, 15(24), 3508; https://doi.org/10.3390/ani15243508 - 5 Dec 2025

Abstract

Pig production plays a vital role in global agriculture, serving as a key source of animal protein and contributing significantly to food security, rural livelihoods, and national economies [...] Full article

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

24 pages, 5160 KB

Open AccessArticle

Using Satellite Remote Sensing to Estimate Rangeland Carrying Capacity for Sustainable Management of the Marismeño Horse in Doñana National Park, Spain

by Emilio Ramírez-Juidias, Ángel Díaz de la Serna-Moreno and Manuel Delgado-Pertíñez

Animals 2025, 15(24), 3507; https://doi.org/10.3390/ani15243507 - 5 Dec 2025

Abstract

Rangeland degradation poses a serious challenge for the sustainable management of free-ranging livestock in Mediterranean wetlands. In Doñana National Park, Spain, the endangered Marismeño horse depends exclusively on natural forage, making it essential to monitor vegetation productivity and grazing suitability under increasing climate [...] Read more.

Rangeland degradation poses a serious challenge for the sustainable management of free-ranging livestock in Mediterranean wetlands. In Doñana National Park, Spain, the endangered Marismeño horse depends exclusively on natural forage, making it essential to monitor vegetation productivity and grazing suitability under increasing climate variability. This study presents a satellite-based assessment of rangeland carrying capacity to support the adaptive management of this iconic breed. A six-year time series (2015–2020) of 1242 images from Landsat 8 OLI/TIRS and Sentinel-2 (L1C/L2A) was processed using ILWIS and Python-based workflows to derive vegetation indices (GNDVI, NDMI) and model aboveground biomass, forage energy, and grazing pressure across five grazing units. Results revealed strong seasonal cycles, with biomass and nutritive value peaking in spring and declining sharply in summer. Ecotonal zones such as La Vera y Sotos acted as crucial refuges during drought-induced resource shortages. The harmonized multi-sensor approach demonstrated high reliability for mapping forage dynamics and assessing carrying capacity at fine scales. This remote sensing framework offers an effective, scalable tool for sustainable livestock management in Doñana, directly supporting biodiversity conservation and the long-term resilience of Mediterranean rangeland ecosystems. Full article

(This article belongs to the Section Equids)

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26 pages, 5462 KB

Open AccessFeature PaperReview

Diagnostic Imaging Features of Mammary Gland Tumors in Dogs and Cats

by Marisa Esteves-Monteiro, Joana Santos, Ana Patrícia Fontes-Sousa and Cláudia S. Baptista

Animals 2025, 15(24), 3506; https://doi.org/10.3390/ani15243506 - 5 Dec 2025

Abstract

Mammary gland tumors (MGT) are the most common neoplasms in intact female dogs and represent the third most frequent tumor type in female cats. Imaging plays a central role in the diagnosis, staging, and follow-up of these neoplasms. This review summarizes the diagnostic [...] Read more.

Mammary gland tumors (MGT) are the most common neoplasms in intact female dogs and represent the third most frequent tumor type in female cats. Imaging plays a central role in the diagnosis, staging, and follow-up of these neoplasms. This review summarizes the diagnostic imaging features of canine and feline MGT and the advantages and limitations of each modality. Radiography and computed tomography (CT) are mainly used to detect pulmonary and abdominal metastases, while ultrasonography provides key information about primary tumor size, margins, echogenicity, vascularization, and elasticity. Advanced ultrasound techniques, such as Doppler, contrast-enhanced ultrasonography (CEUS), and elastography, improve differentiation between benign and malignant lesions. Magnetic resonance imaging (MRI) offers excellent soft-tissue detail and can assist in preoperative planning, whereas nuclear medicine techniques, including scintigraphy and positron emission tomography (PET), allow functional assessment and the early detection of micrometastases. Although histopathology remains the diagnostic gold standard, imaging is indispensable for characterizing primary lesions, evaluating metastatic spread, guiding sampling, and monitoring therapeutic outcomes in small animal oncology. By integrating and comparing the main imaging modalities applied to canine and feline mammary tumors, this review underscores their complementary roles in improving diagnosis, staging, and therapeutic decision-making in small animal oncology. Full article

(This article belongs to the Special Issue Diagnostic Imaging in Animal Oncology)

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15 pages, 1267 KB

Open AccessArticle

Whole-Genome Sequencing of Dorper × Hu Hybrid Sheep for Screening Selection Signatures Associated with Litter Size

by Liying Qiao, Ke Ma, Quanhong Yao, Siying Zhang, Zhixu Pang, Wannian Wang, Ke Cai and Wenzhong Liu

Animals 2025, 15(23), 3505; https://doi.org/10.3390/ani15233505 - 4 Dec 2025

Abstract

Sheep are an economically important livestock species, and reproductive performance is a key trait affecting productivity. The Dorper × Hu hybrid sheep (DHS), widely bred in China, provides a valuable model for studying the genetic basis of prolificacy. This study aimed to investigate [...] Read more.

Sheep are an economically important livestock species, and reproductive performance is a key trait affecting productivity. The Dorper × Hu hybrid sheep (DHS), widely bred in China, provides a valuable model for studying the genetic basis of prolificacy. This study aimed to investigate the genomic architecture and identify candidate genes associated with high litter size in DHS using whole-genome selective sweep analysis and genome-wide association study (GWAS). A total of 31 DHS individuals with complete reproductive records were sequenced and compared with publicly available genomic data from 20 Hu sheep (HUS) and 10 Dorper sheep (DPS). Population genetic structure and diversity were assessed using phylogenetic trees, principal component analysis (PCA), and ADMIXTURE analysis. To identify key genomic regions associated with litter size, we performed selective sweep analysis between the polytocous and monotocous subpopulations of DHS using multiple methods within a 50 kb sliding window framework, including

F_{S T}

,

θ_{π}

ratio, XP-CLR, and XP-EHH; we also conducted GWAS. DHS exhibited a distinct genetic structure with admixed ancestry and elevated genetic diversity. Genetic diversity analysis showed that DHS retained moderate levels of heterozygosity and polymorphism, comparable to or exceeding those of its parental breeds. Comparative analysis between polytocous and monotocous DHS identified reproduction-associated genes, including MUC1, PLCB4, SIN3A, and ELAVL2, enriched in pathways such as ovarian steroidogenesis, insulin secretion, and circadian entrainment. Furthermore, genome-wide association study (GWAS) identified 140 significant loci (p < 10⁻⁵) associated with reproductive traits. From these, 10 candidate SNPs were selected for validation through single-marker association analysis in 200 DHS individuals, among which two loci—g.88680390 C>A (SLC24A2/MLLT3) and g.18197516 T>C (ABCA1)—showed significant correlations with litter size. These findings enhance our understanding of the genetic basis of prolificacy in DHS and provide valuable molecular markers for genomic selection in sheep-breeding programs. Full article

(This article belongs to the Topic Advances in Molecular Genetics and Breeding of Cattle, Sheep, and Goats)

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26 pages, 6574 KB

Open AccessArticle

A Dietary Supplement Containing Nucleotides, Oligosaccharides, Vitamin E and β-Carotene Promotes Immune Response and Gut Microbiota Changes in Kittens

by Willy Joly, Matthew Harrison, Jeremy Laxalde and Virginie Gaillard

Animals 2025, 15(23), 3504; https://doi.org/10.3390/ani15233504 - 4 Dec 2025

Abstract

Kittens are born with some maternally derived antibodies (MDA) that, together with maternal gut microbiota, bring some form of early defense against pathogens. However, this protection declines and the rapid changes and challenges the kitten typically encounters can adversely affect their health. Nutrition [...] Read more.

Kittens are born with some maternally derived antibodies (MDA) that, together with maternal gut microbiota, bring some form of early defense against pathogens. However, this protection declines and the rapid changes and challenges the kitten typically encounters can adversely affect their health. Nutrition can modulate the immune system and gut microbiota, offering a boost to protection following weaning. This controlled, randomized, double-blinded study tests a dietary supplement containing nucleotides, oligosaccharides, vitamin E and β-carotene in 50 domestic short-haired kittens. Control kittens were fed standard, dry kitten diet whilst test kittens received the same diet with supplement for 52 weeks. Kittens received routine vaccinations and blood samples were collected throughout the study for standard hematology, serum antibodies and cytokine expression. Rectal fecal samples were analyzed for gut microbiota via 16S rRNA gene sequencing. Supplemented kittens showed significantly improved antibody response to Chlamydia vaccine at various timepoints versus controls, with no consistent effects of supplementation on responses to other vaccines. Several cytokines were significantly influenced by the supplement, which also significantly altered gut microbiota diversity. In conclusion, a dietary supplement was shown to promote immune response and gut microbiota changes in kittens and may help to support a healthy transition into adulthood. Full article

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

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26 pages, 3611 KB

Open AccessCase Report

Successful Rescue of a Juvenile Humpback Whale (Megaptera novaeangliae) Trapped Upstream of the Rance Tidal Power Station, Brittany, France

by Oihana Olhasque, Léanne Carpentier, Matthieu Duchemin, Jean-Luc Jung, Cécile Dars, Florian Boucard, Sophie Labrut and Joëlle De Weerdt

Animals 2025, 15(23), 3503; https://doi.org/10.3390/ani15233503 - 4 Dec 2025

Abstract

Rescue operations involving baleen whales trapped in dammed environments are difficult to perform successfully, yet increasingly relevant under growing coastal development. We report on a two-day (9–10 February 2023) rescue of a juvenile humpback whale trapped upstream of the Rance Tidal Power Station [...] Read more.

Rescue operations involving baleen whales trapped in dammed environments are difficult to perform successfully, yet increasingly relevant under growing coastal development. We report on a two-day (9–10 February 2023) rescue of a juvenile humpback whale trapped upstream of the Rance Tidal Power Station (TPS) in Brittany, France, providing rare peer-review evidence on response strategies in highly engineered estuaries. A collaborative, non-invasive strategy was implemented by adjusting water levels and creating artificial tidal currents to prevent the whale from stranding and to guide the individual back to open water. Approximately 100 people were mobilized as part of the rescue operation. This paper describes a detailed spatiotemporal account of the whale’s movements and a chronological record of the actions taken by the rescue team. After several attempts to guide it out, rescue efforts enabled its successful exit from the estuary on the second day of operations, and it was not subsequently reported stranded along the French coast. This case demonstrates the value of rapid multidisciplinary coordination between the French National Stranding Network (composed of marine scientists, veterinarians and local correspondents), local organizations, the local marine biology station, international marine mammal experts, national institutions, authorities and a tidal energy operator. Beyond documenting an unusual event, this paper provides operational lessons, highlighting (i) the adaptative management of a TPS as a guidance tool, (ii) the prioritization of animal welfare and responders’ safety, (iii) the effective public and media management and (iv) the involvement of trained volunteers during the rescue, promoting environmentally responsible behavior. These insights are transferable to other cases to inform future baleen whales rescue protocols in anthropogenic environments. Full article

(This article belongs to the Section Mammals)

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