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Working on the Frontline of Dog Adoption: The Perspectives and Experiences of Animal Shelter Workers in RSPCA Queensland
Anatomic Interactive Atlas of the Loggerhead Sea Turtle (Caretta caretta) Coelomic Cavity
Valorisation of Rabbit Biodiversity for Meat Production: Evaluation of Different Rabbit Genotypes
Applicability of Machine Learning in Behavioural Monitoring of the Red Panda (Ailurus fulgens) in Zoos
Effects of Probiotics Weizmannia faecalis and Bacillus licheniformis on Performance and Carcass Traits of Broilers

Journal Description

Animals

Animals is an international, peer-reviewed, open access journal devoted entirely to animals, including zoology and veterinary sciences, and is published semimonthly online by MDPI. The World Association of Zoos and Aquariums (WAZA), European College of Animal Welfare and Behavioural Medicine (ECAWBM), and Federation of European Laboratory Animal Science Associations (FELASA) are affiliated with Animals and their members receive a discount on the article processing charges.

Open Access— free for readers, with article processing charges (APC) paid by authors or their institutions.
High Visibility: indexed within Scopus, SCIE (Web of Science), PubMed, PMC, Embase, PubAg, AGRIS, Animal Science Database, CAB Abstracts, and other databases.
Journal Rank: JCR - Q1 (Veterinary Sciences) / CiteScore - Q1 (General Veterinary )
Rapid Publication: manuscripts are peer-reviewed and a first decision is provided to authors approximately 17.1 days after submission; acceptance to publication is undertaken in 2.6 days (median values for papers published in this journal in the second half of 2025).
Recognition of Reviewers: reviewers who provide timely, thorough peer-review reports receive vouchers entitling them to a discount on the APC of their next publication in any MDPI journal, in appreciation of the work done.
Companion journals for Animals include: Birds, Ruminants and Zoonotic Diseases.
Journal Cluster of Animal Science: Animals, Arthropoda, Birds, Dairy, Insects, Journal of Zoological and Botanical Gardens, Pets, Poultry, Ruminants and Veterinary Sciences.

Impact Factor: 2.7 (2024); 5-Year Impact Factor: 3.2 (2024)

Imprint Information Journal Flyer Open Access ISSN: 2076-2615

Latest Articles

15 pages, 7563 KB

Open AccessArticle

Notoginsenoside R1 Inhibits Porcine Deltacoronavirus Infection In Vitro by Restoring SERCA2-Mediated Calcium Homeostasis

by Jialu Zhang, Yuqian Liu, Wenzhe Liu, Zhouyuan Wang, Hanlu Wang, Xuejing Xia, Lianci Peng, Tingting Chen and Rendong Fang

Animals 2026, 16(12), 1836; https://doi.org/10.3390/ani16121836 (registering DOI) - 14 Jun 2026

Porcine coronavirus is one of the prevalent enteric coronaviruses in pigs, causing watery diarrhea and even death in suckling piglets and resulting in giant losses to the pig industry. However, effective antiviral strategies against PDCoV remain limited. Notoginsenoside R1 (NG-R1), a saponin extracted [...] Read more.

Porcine coronavirus is one of the prevalent enteric coronaviruses in pigs, causing watery diarrhea and even death in suckling piglets and resulting in giant losses to the pig industry. However, effective antiviral strategies against PDCoV remain limited. Notoginsenoside R1 (NG-R1), a saponin extracted from Panax notoginseng, exhibits diverse bioactivities, but its antiviral potential has not been fully characterized. Herein, we systematically investigated the anti-PDCoV effect of NG-R1 and its underlying mechanism. NG-R1 showed no cytotoxic effect on LLC-PK1 cells and exerted antiviral ability against PDCoV infection through targeting the whole life cycle of the virus. In addition, network pharmacology analysis identified calcium signaling as a potentially relevant pathway involved in the antiviral activity of NG-R1. Further data demonstrated that PDCoV infection disrupted intracellular calcium homeostasis, whereas NG-R1 treatment partially restored calcium balance and attenuated endoplasmic reticulum (ER) stress. Moreover, NG-R1 modulated the expression of SERCA2, a key regulator of ER calcium transport. Thapsigargin, an inhibitor of SERCA2, showed similar antiviral capacity to NG-R1. Collectively, our findings suggest that NG-R1 exerts antiviral activity against PDCoV, potentially through regulation of calcium homeostasis mediated by SERCA2. This study provides a theoretical basis for the development of novel antiviral agents targeting calcium signaling pathways. Full article

(This article belongs to the Special Issue Diarrheal Diseases in Piglets: Pathogenic Mechanism, Prevention and Control)

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19 pages, 281 KB

Open AccessArticle

Enhancing Rehydrated Rice Husk as Ruminant Feed via Silage Additives: An In Vitro Study

by Chatchai Kaewpila, Julasinee Maensathit, Pairote Patarapreecha and Waroon Khota

Animals 2026, 16(12), 1835; https://doi.org/10.3390/ani16121835 (registering DOI) - 14 Jun 2026

Rice husk is an abundant agricultural by-product with limited use in ruminant feeding due to its high lignocellulosic content and low digestibility. This study compared biological, enzymatic, and chemical additive strategies for improving the ensiling characteristics, chemical composition, and in vitro rumen fermentation [...] Read more.

Rice husk is an abundant agricultural by-product with limited use in ruminant feeding due to its high lignocellulosic content and low digestibility. This study compared biological, enzymatic, and chemical additive strategies for improving the ensiling characteristics, chemical composition, and in vitro rumen fermentation of rehydrated rice husk. The experiment was conducted using a completely randomized design with seven treatments: Control (no additive); molasses + Lacticaseibacillus casei TH14 (MB); Acremonium cellulase (AC); laccase (LC); AC + LC; AC + LC + MB; and chemical treatment (CM). After 30 days of ensiling, CM reduced fiber contents and increased in vitro dry matter digestibility (IVDMD) and total volatile fatty acid concentrations (p < 0.05). Among the biological additives, MB and AC + LC + MB significantly reduced NDF and improved ensiling quality and IVDMD (p < 0.05). Notably, AC + LC + MB resulted in the lowest methane emission intensity (44.07 mg/g IVDMD). Although CM, which was included as a chemical-disruption benchmark, was the most effective approach for substrate solubilization, AC + LC + MB improved fermentation characteristics, in vitro digestibility, and methane emission intensity per unit of digested dry matter under the present in vitro conditions. Full article

(This article belongs to the Special Issue Local Feed Resources in Ruminants Nutrition)

15 pages, 463 KB

Open AccessArticle

Phenotypic Variability of Wool Traits in the Gentile di Puglia Sheep Breed: Implications for Conservation and Sustainable Management

by Maria Gabriela Molina, Grazia Bramante, Claudia Pierini, Medhat S. Saleh, Antonietta D’Onghia, Silverio Grande, Giuseppe Mangini, Silvia Bruno, Virginia Devoto, Elena Ciani, Francesca Maria Sarti, Pasquale De Palo and Vincenzo Landi

Animals 2026, 16(12), 1834; https://doi.org/10.3390/ani16121834 (registering DOI) - 14 Jun 2026

The Gentile di Puglia sheep breed represents an important component of Southern Italy’s livestock biodiversity, yet its wool traits have not been systematically evaluated for several decades. This study aimed to provide an updated phenotypic assessment of key wool characteristics and to explore [...] Read more.

The Gentile di Puglia sheep breed represents an important component of Southern Italy’s livestock biodiversity, yet its wool traits have not been systematically evaluated for several decades. This study aimed to provide an updated phenotypic assessment of key wool characteristics and to explore the structure of their variability across production contexts. Data were collected from 3089 animals from nine farms over two consecutive years and included fibre diameter, staple length, coefficient of variation in fibre diameter, and clean yield. Descriptive statistics, correlation analysis, principal component analysis, and mixed-effects models were applied to evaluate phenotypic patterns and partition sources of variability. Fibre diameter and staple length showed moderate variability, with most variation occurring within farm-year units, whereas clean yield exhibited a stronger dependence on farm-year conditions. Correlations among traits were generally weak, indicating a high degree of independence. The multivariate analysis revealed a diffuse structure, with no dominant axis explaining most of the variability. Variance component analysis confirmed that production context had a greater influence on clean yield than on fibre-related traits. These results highlight the importance of both conservation and management strategies. The observed variability may provide useful opportunities for future breeding and conservation strategies, while the sensitivity of clean yield to environmental and management factors underscores the need for improved handling practices. Overall, the Gentile di Puglia breed retains a flexible phenotypic profile that supports its conservation and potential valorisation in sustainable production systems. Full article

(This article belongs to the Section Small Ruminants)

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16 pages, 4460 KB

Open AccessArticle

Epimedii Folium Supplementation Improves Semen Quality, Hormonal Profile, and Immune Function by Modulating Gut Microbiota and Seminal Metabolites in Aged Boars

by Bin Ran, Shengxin Luo, Chenxi Zhou, Long Wen, Junjie Wu, Yunxiang Zhao, Xiaoping Zhu, Zhili Li and Mengjie Liu

Animals 2026, 16(12), 1833; https://doi.org/10.3390/ani16121833 (registering DOI) - 14 Jun 2026

This study aimed to investigate the effects of dietary Herba Epimedii Folium (HEF) supplementation on semen quality, reproductive hormones, immune parameters, gut microbiota, and seminal plasma metabolites in aged boars, and to evaluate its potential for extending their reproductive lifespan. A total of [...] Read more.

This study aimed to investigate the effects of dietary Herba Epimedii Folium (HEF) supplementation on semen quality, reproductive hormones, immune parameters, gut microbiota, and seminal plasma metabolites in aged boars, and to evaluate its potential for extending their reproductive lifespan. A total of 18 Bama boars (approximately 3 years of age) were randomly assigned to three groups (n = 6 per group). The control group received a basal diet, while the treatment groups were fed the basal diet supplemented with 3 g/kg or 5 g/kg of HEF for 8 weeks. The results showed that adding HEF to the diet of aged boars increased the motility and concentration of their sperm and reduced the proportion of abnormal sperm. Treatment with 3 g/kg HEF increased serum LH and IgG levels, whereas the 5 g/kg dose elevated IgA levels in both serum and seminal plasma, as well as IgG levels in seminal plasma. Furthermore, 16S rRNA sequencing revealed that dietary HEF supplementation reduced the relative abundance of Streptococcus and Oscillospiraceae UCG-002 in the gut of aged boars. PICRUSt2 analysis predicted that pathways involved in lysine biosynthesis, arginine and proline metabolism, glycine, serine and threonine metabolism, and amino acid-related enzymes were enriched in the HEF treatment group. Semen metabolite profiling showed that the HEF treatment enriched several key metabolites, including 5-hydroxytryptophan, acetylcarnitine, tretinoin, methyltestosterone, prostaglandin A3, and prostaglandin B2. Spearman correlation analysis revealed a negative association between Streptococcus abundance and sperm motility, whereas acetylcarnitine, 5-hydroxytryptophan, and prostaglandin A3 were positively associated with motility. Furthermore, 5-hydroxytryptophan levels were positively linked to both sperm concentration and serum LH. In summary, our study demonstrates that Epimedii Folium may enhance the semen quality of aged Bama boars by improving the intestinal microbiota and the metabolic profile of seminal plasma. These findings may offer a theoretical basis for optimizing reproduction and conserving germplasm resources in aged Bama miniature pigs. Full article

(This article belongs to the Special Issue The Role of Reproductive Microbiota in Animal Fertility and Reproductive Health)

17 pages, 715 KB

Open AccessReview

Neuroimmune Mechanisms in Equine Asthma: Primary Inflammatory Triggers, Neuroimmune Modulation and Chronic Airway Remodelling

by Małgorzata Wierzbicka, Aleksandra Samsel and Marta Siemieniuch-Tartanus

Animals 2026, 16(12), 1832; https://doi.org/10.3390/ani16121832 (registering DOI) - 14 Jun 2026

Equine asthma is a chronic inflammatory disease of the lower respiratory tract, primarily associated with inhalation of organic dust, microbial particles and environmental aeroantigens. Although the inflammatory and immunological mechanisms underlying equine asthma have been extensively investigated, the potential contribution of neuroimmune pathways [...] Read more.

Equine asthma is a chronic inflammatory disease of the lower respiratory tract, primarily associated with inhalation of organic dust, microbial particles and environmental aeroantigens. Although the inflammatory and immunological mechanisms underlying equine asthma have been extensively investigated, the potential contribution of neuroimmune pathways remains poorly understood. In humans and rodent models, asthma is increasingly recognised as a disorder involving complex bidirectional interactions between the nervous and immune systems. Sensory nerve activation, neuropeptide release, autonomic dysregulation and neuronal remodelling contribute to bronchoconstriction, airway hyperresponsiveness, mucus hypersecretion and chronic airway remodelling. This review summarises current knowledge of the neuroimmune mechanisms involved in asthma, with particular emphasis on comparative aspects across humans, rodents and horses. Literature searches were conducted using the PubMed database, focusing on studies investigating neurogenic inflammation, airway innervation, neuropeptides, transient receptor potential channels and neuronal remodelling in asthma and chronic airway disease. Existing equine evidence indicates the presence of substance P- and calcitonin gene-related peptide-immunoreactive nerve fibres in the equine airways, increased neurokinin-mediated bronchoconstriction in severe equine asthma, and enhanced airway innervation in affected horses. However, compared with human and rodent studies, horse-specific data remain extremely limited. Current evidence suggests that neuroimmune pathways are unlikely to be the primary initiating mechanism of equine asthma, but may act as important modulators of chronic airway dysfunction and disease progression. The marked scarcity of equine studies investigating neuroimmune signalling represents a major knowledge gap and highlights an important direction for future research in equine respiratory medicine. Full article

(This article belongs to the Special Issue Equine Asthma: From Pathogenesis to Therapy)

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22 pages, 1729 KB

Open AccessReview

Retinoic Acid Signaling in Male Reproductive Biology: From Germ Cell Regulation to Contraceptive Innovation Within a One Health Framework

by Vanmathy Kasimanickam and Ramanathan Kasimanickam

Animals 2026, 16(12), 1831; https://doi.org/10.3390/ani16121831 (registering DOI) - 14 Jun 2026

Spermatogenesis is a highly coordinated biological process in which diploid spermatogonia undergo mitotic expansion, meiotic division, and terminal differentiation into haploid spermatozoa. This process is tightly regulated by intrinsic germ cell programs and extrinsic signals from Sertoli cells within the seminiferous epithelium. Among [...] Read more.

Spermatogenesis is a highly coordinated biological process in which diploid spermatogonia undergo mitotic expansion, meiotic division, and terminal differentiation into haploid spermatozoa. This process is tightly regulated by intrinsic germ cell programs and extrinsic signals from Sertoli cells within the seminiferous epithelium. Among the signaling pathways governing male germ cell development, all-trans retinoic acid (RA), a bioactive metabolite of vitamin A, has emerged as a master regulator of meiotic initiation and spermatogonial differentiation in mammals. RA functions through nuclear retinoic acid receptors (RARs) and retinoid X receptors (RXRs), which regulate transcriptional networks essential for germ cell progression, including the activation of Stimulated by Retinoic Acid 8 (STRA8), a key determinant of meiotic entry. Intratesticular RA homeostasis is maintained by a balance between synthesis via aldehyde dehydrogenase (ALDH) enzymes and degradation by cytochrome P450 family 26 (CYP26) enzymes, ensuring precise temporal and spatial control of germ cell development. While rodent models have defined core mechanisms of RA signaling, the canine testis provides a valuable comparative and translational system due to its physiological similarity to human spermatogenesis and relevance to reproductive management. Recent studies highlight conserved RA signaling pathways in dogs, including receptor-mediated transcriptional regulation, feedback control of RA metabolism, and post-transcriptional modulation via microRNAs. Importantly, pharmacological manipulation of RA signaling can reversibly disrupt spermatogenesis, supporting its potential applications in non-hormonal male contraception. This review integrates molecular, developmental, pharmacological, and comparative evidence and presents RA signaling as a central regulatory axis of spermatogenesis with important translational applications. Full article

(This article belongs to the Section Animal Reproduction)

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15 pages, 1287 KB

Open AccessArticle

Perceived Rabbit Care Knowledge Does Not Consistently Align with Owner-Reported Husbandry Practices: A Survey of Pet Rabbit Owners in Croatia

by Magdalena Neva Oelsner, Ivana Sabolek, Jurica Novak and Gordana Gregurić Gračner

Animals 2026, 16(12), 1830; https://doi.org/10.3390/ani16121830 (registering DOI) - 14 Jun 2026

The welfare of pet rabbits is frequently compromised despite increasing recognition of their complex behavioural and physiological needs. Although owner knowledge is considered an important determinant of animal care, the relationship between perceived knowledge and actual husbandry practices remains poorly understood. This study [...] Read more.

The welfare of pet rabbits is frequently compromised despite increasing recognition of their complex behavioural and physiological needs. Although owner knowledge is considered an important determinant of animal care, the relationship between perceived knowledge and actual husbandry practices remains poorly understood. This study aimed to (i) describe husbandry practices and owner-reported knowledge among pet rabbit owners in Croatia and (ii) assess whether higher perceived knowledge is associated with greater alignment with recommended husbandry practices. Data were collected using an online questionnaire distributed to rabbit owners (n = 529). The study assessed owner-reported husbandry practices and perceived knowledge but did not include direct assessments of rabbit health or welfare. Although most respondents reported high levels of knowledge regarding rabbit care (85.6%), this was not consistently reflected in implemented husbandry practices. Higher perceived knowledge was significantly associated (p < 0.05) with environmental enrichment, housing type, and cleaning frequency, but these associations were generally weak to moderate. In contrast, perceived knowledge showed little or no association with several key welfare-related practices, including social housing and access to free movement, while only limited associations were observed for feeding practices. These findings reveal a substantial knowledge–practice gap, suggesting that confidence in rabbit care knowledge does not necessarily translate into appropriate husbandry. Therefore, educational approaches focused solely on information provision may be insufficient to ensure the implementation of recommended husbandry practices. Full article

(This article belongs to the Section Animal Welfare)

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14 pages, 1239 KB

Open AccessCommunication

A Functional SNP in the AMH Gene Is Associated with Litter Size in Dazu Black Goats

by Lei Wang, Yiyu Zhang, Qiuyan Li, Yingping Xia, Cong Zhou, Ying Gong, Xinyang Yao, Xinyan Zhu, Zhen Wang, Ke Zhang, Xiaowei Sun, Dejun Xu and Zhongquan Zhao

Animals 2026, 16(12), 1829; https://doi.org/10.3390/ani16121829 (registering DOI) - 14 Jun 2026

AMH plays a critical role in ovarian follicular development and may influence reproductive performance. However, its genetic variation and association with litter size in goats remain unclear. In this study, three single-nucleotide polymorphisms (SNPs) in the AMH gene (g.89169447C>A, g.89169684G>A, and g.89170008T>C) were [...] Read more.

AMH plays a critical role in ovarian follicular development and may influence reproductive performance. However, its genetic variation and association with litter size in goats remain unclear. In this study, three single-nucleotide polymorphisms (SNPs) in the AMH gene (g.89169447C>A, g.89169684G>A, and g.89170008T>C) were identified in Dazu Black goats. Association analysis showed that the g.89169447C>A locus was significantly associated with litter size at first parity (p < 0.05), while the g.89169684G>A locus was significantly associated with the average litter size across the first three parities (p < 0.05). Functional assays in granulosa cells indicated that the g.89169684G>A variant altered the expression of proliferation and apoptosis-related genes. These results suggest that the g.89169684G>A variant may serve as a potential molecular marker for improving reproductive performance in goats. Full article

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

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19 pages, 4790 KB

Open AccessArticle

Biphasic Responses of Porcine Oocytes to Metformin: Concentration-Dependent AMPK Activation and Nrf2-Mediated Antioxidant Regulation

by Junyu Wang, Min Li, Yaqi Zhou, Fuyin Fu, Feng Liu, Jinghe Tan, Mingjiu Luo and Shuai Gong

Animals 2026, 16(12), 1828; https://doi.org/10.3390/ani16121828 (registering DOI) - 13 Jun 2026

Metformin (MET) plays crucial regulatory roles in mammalian oocyte meiosis, yet the concentration-dependent biphasic impacts of MET on porcine oocyte in vitro maturation (IVM) and the related molecular mechanisms remain poorly clarified. This study aimed to explore the distinct effects and underlying pathways [...] Read more.

Metformin (MET) plays crucial regulatory roles in mammalian oocyte meiosis, yet the concentration-dependent biphasic impacts of MET on porcine oocyte in vitro maturation (IVM) and the related molecular mechanisms remain poorly clarified. This study aimed to explore the distinct effects and underlying pathways of low- and high-dose MET in porcine oocytes. Different concentrations of MET (0, 7.5, 15, 30, 150, and 300 μM) were supplemented during oocyte IVM, with phenotypic detection, untargeted metabolomic analysis, and Nrf2 inhibitor (ML385) intervention performed for mechanism exploration. Results showed that 15 μM low-dose MET facilitated oocyte maturation, mitochondrial function and redox balance, while 300 μM high-dose MET caused obvious developmental damage. Mechanistically, low-dose MET triggered noncanonical AMPK activation independent of the AMP/ATP ratio and enhanced AMPK–Nrf2 antioxidant signaling, whereas high-dose MET induced energy stress and oxidative injury via inhibiting mitochondrial complex I. Blockade of Nrf2 further abolished the protective effects of low-dose MET. Collectively, this finding illustrates the biphasic actions of MET on porcine oocytes and provides a theoretical reference for optimizing porcine in vitro embryo production. Full article

(This article belongs to the Special Issue Advances in Pig Reproductive Physiology)

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18 pages, 2236 KB

Open AccessArticle

Dietary Guanidinoacetic Acid Improves Meat Tenderness and Antioxidant Capacity in Rabbits via Modulating Muscle Fiber Characteristics and Fat Metabolism

by Yanhui Liang, Xi Chen, Xiaoyu Fan, Yingmei Zhang, Shengnan Wang, Xiaojia Wu, Yingle Wei, Changmao Wei, Yichen Lin, Qinghua Liu and Changchuan Ye

Animals 2026, 16(12), 1827; https://doi.org/10.3390/ani16121827 (registering DOI) - 12 Jun 2026

As the direct biosynthetic precursor of creatine, guanidinoacetic acid (GAA) exerts a pivotal regulatory role in energy homeostasis and protein metabolism. Rabbit meat has garnered increasing global recognition as a healthy food source, characterized by its outstanding high-protein and low-fat nutritional profile. Accordingly, [...] Read more.

As the direct biosynthetic precursor of creatine, guanidinoacetic acid (GAA) exerts a pivotal regulatory role in energy homeostasis and protein metabolism. Rabbit meat has garnered increasing global recognition as a healthy food source, characterized by its outstanding high-protein and low-fat nutritional profile. Accordingly, the optimization of rabbit meat quality has attracted growing attention from both consumers and animal production practitioners. In the present study, we evaluated the impacts of dietary GAA supplementation on meat quality traits, in vivo antioxidant capacity, muscle fiber characteristics, and fatty acid metabolism in New Zealand white rabbits. A total of 960 male New Zealand white rabbits were assigned to two age groups: 40-day-old group and 60-day-old group (40 ± 2 days, 1.19 ± 0.09 kg; 60 ± 2 days, 1.82 ± 0.15 kg). Within each age group, rabbits were randomly allocated to a control diet or a diet supplemented with 100 mg/kg GAA (CON-40, GAA-40, CON-60, GAA-60). After a 45-day feeding period, two-way ANOVA revealed that GAA supplementation significantly reduced shear force (p < 0.01, diet main effect) and muscle fiber density (p < 0.01, diet main effect), with an age-dependent effect on shear force (age × diet interaction, p < 0.05). Moreover, GAA enhanced systemic antioxidant capacity, as indicated by increased serum superoxide dismutase (SOD) activity (p < 0.01) and total antioxidant capacity (T-AOC) (p < 0.05), while no significant effect on malondialdehyde (MDA) was detected under the current experimental conditions. GAA also regulated the expression of lipid metabolism-related genes (FAS, HSL, ACC) in intramuscular and perirenal fat, indicating its regulatory effect on fatty acid metabolism. In conclusion, dietary GAA supplementation improves rabbit meat tenderness and antioxidant capacity, with no negative effects on growth performance. These findings confirm that GAA has the potential to serve as a nutritional strategy to improve rabbit meat quality, supporting the development of rabbit meat as a functional food for human consumption. Full article

(This article belongs to the Special Issue Effect of Feeding Strategies and Nutritional Management on the Quality of Animal Products)

25 pages, 1819 KB

Open AccessArticle

Observations on Anti-Predator Defense Behavior in Feral Horses in Venezuela

by Lucy Rees and Emily Kieson

Animals 2026, 16(12), 1826; https://doi.org/10.3390/ani16121826 (registering DOI) - 12 Jun 2026

Apart from vigilance and flight, anti-predator defense behavior in horses has not been well documented despite its importance during natural selection. In this study, observations of a feral herd (around 140) of Venezuelan horses sympatric with puma and jaguar divided such defense into [...] Read more.

Apart from vigilance and flight, anti-predator defense behavior in horses has not been well documented despite its importance during natural selection. In this study, observations of a feral herd (around 140) of Venezuelan horses sympatric with puma and jaguar divided such defense into precaution and reaction. Group living and the avoidance of danger areas are precautionary measures enhanced by the stallion’s vigilance and his actions to keep small foals with the band. Reactions to perceived threats comprise communication of alarm; bunching, or cohesion, as a primary response; massed flight following self-organizing principles; and reassembly of bands. Stallions usually initiated this behavioral process. Stallions’ initial reactions to perceived threats were “investigation”, “move away”, “run away”, and “stampede”, and resulting herd behavior was categorized into 27 responses. Data analysis through Observation Oriented Modeling indicated that each category of initial stallion response to perceived threats was associated with a recurring pattern of subsequent herd behavior. Prominent behaviors enhanced cohesion and synchrony, as well as velocity and direction matching. A fourth observed category was the cohesive “run to band” of a startled outlying member, in which the individual’s alarm might transmit to the band or the band’s calm transmit to the individual. The results emphasize the importance of communication, social cohesion, and synchronous action in times of perceived threats, their continuous practice during maintenance activities, and the social needs and understanding management of domestic horses. Full article

(This article belongs to the Section Equids)

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21 pages, 1370 KB

Open AccessArticle

Transcriptomics and Metabolomics Signatures of Fat Deposition Following Orchiectomy in Yak

by Lin Xiong, Jie Pei, Qianyun Ge, Zhiqiang Ding, Yandong Kang, Chao Chen, Ruichao Wei and Xian Guo

Animals 2026, 16(12), 1825; https://doi.org/10.3390/ani16121825 (registering DOI) - 12 Jun 2026

Fat deposition plays an important role in yak metabolism, reproduction, and meat quality, and male yaks are often castrated to facilitate management and improve production performance. The effect of castration on the characteristics of fat deposition in male yaks and the molecular mechanisms [...] Read more.

Fat deposition plays an important role in yak metabolism, reproduction, and meat quality, and male yaks are often castrated to facilitate management and improve production performance. The effect of castration on the characteristics of fat deposition in male yaks and the molecular mechanisms of action was explored in this study. The subcutaneous fat thickness in castrated and common male yaks was measured, further the content of fatty acids in yak subcutaneous fat was detected using gas chromatography-mass spectrometer (GC-MS); the transcriptome, metabolome in the yak subcutaneous fat were detected using mRNA-Sequencing, ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), respectively; the integrative analyses of differentially expressed genes (DEGs), different metabolites (DMs), fatty acids and fat thickness were carried out. The results showed that castration can strengthen the ability of fat deposition and improve the content of fatty acids, especially PUFAs, in male yaks, and both transcriptome and metabolome were significantly different between castrated male yaks and common male yaks. The effect of castration on the male yak fat deposition was closely related to the PPAR signaling pathway, citrate cycle, and insulin resistance. Data suggests that FASN, ACACA, AGPAT2, ACLY, ACSL5, SCD, GSK3B, and SLC2A4 may be the crucial control genes for the fat amount in yaks, and that FADS2, LPL, and ACSL4 may be the crucial control genes for the polyunsaturated fatty acids (PUFAs) content in yak adipose tissue. Further functional studies will be conducted to determine the specific role of each gene in regulating fat deposition and fatty acid composition in yaks. Full article

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

19 pages, 1226 KB

Open AccessArticle

Dry Matter Intake Prediction Models: Evaluation Across Energy-Corrected Milk and Lactation-Stage Classes in Holstein Cows

by Ugur Serbester, Ahmet Gorkem Aydoner, Poyraz Yasar Bozkaya and Zeynel Cebeci

Animals 2026, 16(12), 1824; https://doi.org/10.3390/ani16121824 (registering DOI) - 12 Jun 2026

Accurate prediction of dry matter intake (DMI) is essential for ration formulation, nutrient supply, and evaluation of production efficiency in lactating dairy cows. Several DMI prediction models are currently used, but most comparative studies have emphasized overall accuracy rather than whether model bias [...] Read more.

Accurate prediction of dry matter intake (DMI) is essential for ration formulation, nutrient supply, and evaluation of production efficiency in lactating dairy cows. Several DMI prediction models are currently used, but most comparative studies have emphasized overall accuracy rather than whether model bias changes across biologically relevant production contexts. The objective of this study was to evaluate the context-dependent bias of widely used DMI prediction models in lactating dairy cows across classes of energy-corrected milk (ECM) and lactation stage. A literature-derived database was assembled from 135 studies consisting of 436 treatments from 6985 Holstein cows, reporting observed DMI and the variables required to implement five prediction models and evaluate their prediction error (PE): NRC2001, the Cornell Net Carbohydrate and Protein System (CNCPS), NASEM2021, Agroscope2021, and GfE2023. PE was calculated as predicted DMI minus observed DMI, such that positive values indicated overprediction and negative values indicated underprediction. Observations were classified according to ECM and days in milk (DIM). Mixed models were fitted separately for the ECM class and the lactation-stage class, with the study fitted as a random effect. PE differed among models, and the pattern of bias depended on both the ECM and the lactation-stage classes. The interaction between the ECM class and the model was significant, indicating that productive level modified model bias. The interaction between lactation-stage class and model was also significant and more pronounced, indicating marked changes in model bias across lactation stages. Across classes, NASEM2021 generally remained closest to zero, whereas GfE2023 and CNCPS showed more negative PE values in most contexts. Agroscope2021 showed a more context-sensitive pattern, and NRC2001 remained comparatively moderate across several classes. These findings indicate that the evaluation of DMI prediction models based only on global mean bias may conceal an important biological structure in PE. Context-specific evaluation, particularly across the lactation stage, may provide a more informative basis for selecting DMI prediction models for research and practical ration formulation. Full article

(This article belongs to the Section Animal Nutrition)

18 pages, 28097 KB

Open AccessArticle

Percutaneous Cementoplasty as a Monotherapy in the Treatment of Appendicular Osteosarcoma in Ten Dogs

by Sandra L. MacArthur, Kevin A. Drygas, Daniel D. Lewis, James C. Colee and Aquilino Villamonte-Chevalier

Animals 2026, 16(12), 1823; https://doi.org/10.3390/ani16121823 (registering DOI) - 12 Jun 2026

This prospective case series evaluated the short-term outcomes following percutaneous cementoplasty as the sole palliative treatment for appendicular osteosarcoma in 10 dogs. Synthetic self-hardening calcium phosphate bone substitute was injected into the osseous defect under fluoroscopic guidance after curettage of the bone tumor. [...] Read more.

This prospective case series evaluated the short-term outcomes following percutaneous cementoplasty as the sole palliative treatment for appendicular osteosarcoma in 10 dogs. Synthetic self-hardening calcium phosphate bone substitute was injected into the osseous defect under fluoroscopic guidance after curettage of the bone tumor. Clinician assessment included a numerical rating score for lameness, offloading, and ease of lifting the contralateral limb as well as the 4A-VET postoperative pain scale. Owner assessment was obtained using three descriptive questionnaires, the Helsinki Chronic Pain Index (HCPI), the Canine Brief Pain Inventory (CBPI) and the Canine Symptom Assessment Scale (CSAS). Measures were recorded preoperatively and at 2, 4, 8, and 12 weeks following surgery. Early improvement in the 4A-Vet score was noted at the 2-, 4-, 8-, and 12-week time points for all major pain and function metrics. Similarly, the CBPI pain severity and interference scores demonstrated early postoperative improvement during the 2- and 4-week time points with partial attenuation by 8 and 12 weeks. Panting, difficulty sleeping, whining/moaning, and lack of appetite were significantly reduced when assessed via the CSAS. Cementoplasty as a monotherapy, affording early pain relief and improved structural integrity, supports its role as a palliative limb-preserving option for dogs unable to undergo amputation. Full article

(This article belongs to the Special Issue Advances in Small Animal Surgical Oncology)

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14 pages, 1258 KB

Open AccessCommunication

ZooMonitor Community: A Tool for Evaluating Population-Level Behavior Questions

by Jason D. Wark, Nancy Kim Pham and John E. Andrews

Animals 2026, 16(12), 1822; https://doi.org/10.3390/ani16121822 (registering DOI) - 12 Jun 2026

From plains zebras to zebra sharks, modern, accredited zoos and aquariums cooperate on the breeding and transfer of individuals from diverse species with unique needs. As a result, the housing and husbandry of a species varies across organizations. The degree to which these [...] Read more.

From plains zebras to zebra sharks, modern, accredited zoos and aquariums cooperate on the breeding and transfer of individuals from diverse species with unique needs. As a result, the housing and husbandry of a species varies across organizations. The degree to which these practices align with a species’ needs can influence well-being and reproduction. As a key measure of well-being, animal behavior is often the most practical and useful tool for evaluating the impact of housing and husbandry on animals. However, studies conducted at a single organization can be limited in their ability to explore these factors and a multi-institutional approach is needed. Recently, Lincoln Park Zoo launched the ZooMonitor Community, an expansion of the ZooMonitor behavior recording platform. This new collaboration tool is freely available to all zoos and aquariums to facilitate multi-institutional data sharing and project management. Here, we provide an overview of the ZooMonitor Community tool and highlight several recent Community projects on diverse taxa, including giraffes, Asian small-clawed otters, common seadragons, and southern ground hornbills. We hope the ZooMonitor Community can provide population managers with an accessible tool to address broad, species-level questions on care, well-being, and reproduction that can enhance the lives of animals in managed settings. Full article

(This article belongs to the Special Issue Influence of Social Behaviours and Population on Conservation Breeding Programmes and Zoo Animal Welfare)

15 pages, 1160 KB

Open AccessArticle

Ampicillin Depletion and Withdrawal Period in Broilers: Tissue Residue Analysis After Intramuscular Administration

by Paula Cortés, Maximiliano Castillo, Katherine Codoceo Valenzuela, Kevin Manríquez González, Belén Pinto, Ekaterina Pokrant, Aldo Maddaleno, Sebastián Zavala, Andrés Flores and Javiera Cornejo

Animals 2026, 16(12), 1821; https://doi.org/10.3390/ani16121821 (registering DOI) - 12 Jun 2026

Ampicillin residues in animal-derived foods may cause allergic reactions and promote antimicrobial resistance in consumers; however, data on residue behavior in poultry remain limited. This study aimed to evaluate the depletion of ampicillin in muscle and skin plus fat of broiler chickens. Thirty [...] Read more.

Ampicillin residues in animal-derived foods may cause allergic reactions and promote antimicrobial resistance in consumers; however, data on residue behavior in poultry remain limited. This study aimed to evaluate the depletion of ampicillin in muscle and skin plus fat of broiler chickens. Thirty birds were treated with ampicillin intramuscularly (20 mg kg⁻¹ every 24 h for three days) and sacrificed at 0.5, 1, 2, 5, and 9 days post-administration. Samples were analyzed by liquid chromatography coupled with tandem mass spectrometry, a method successfully validated according to Commission Implementing Regulation (EU) 2021/808, VICH GL49 and GL2. Quantification was performed by linear regression from matrix-matched calibration curves. Residue depletion was evaluated following the European Medicines Agency guidelines. Ampicillin residues in muscle were detected only during the first 24 h post-administration (6.50–8.48 µg kg⁻¹). Residues in skin plus fat remained detectable until day 5 post-administration (6.87–59.88 µg kg⁻¹). Based on this, the withdrawal period calculated for skin plus fat was 9 days considering EU maximum residue limit (MRL) and 19 days considering method limit of quantification, with 95% confidence. These results provide critical data on ampicillin residue kinetics under controlled experimental conditions, supporting risk assessments and the establishment of MRLs in broiler chickens by the Codex Alimentarius. Full article

(This article belongs to the Special Issue Pharmacodynamics and Pharmacokinetics of Veterinary Drug Residues)

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17 pages, 1270 KB

Open AccessArticle

Rapid Point-of-Care Detection of Dirofilaria immitis and Dirofilaria repens in Canine Blood Using Two Direct Closed-Tube LAMP Assays

by Zsófia Bujtor, Tünde Földvári, Csaba Pribenszky, Ákos Jerzsele and Petra Zenke

Animals 2026, 16(12), 1820; https://doi.org/10.3390/ani16121820 (registering DOI) - 12 Jun 2026

Canine dirofilariasis, caused by Dirofilaria immitis and Dirofilaria repens, is an emerging vector-borne disease of increasing veterinary and zoonotic importance. Rapid and species-specific detection is essential for effective clinical management and epidemiological surveillance. This study aimed to develop and diagnostically evaluate two [...] Read more.

Canine dirofilariasis, caused by Dirofilaria immitis and Dirofilaria repens, is an emerging vector-borne disease of increasing veterinary and zoonotic importance. Rapid and species-specific detection is essential for effective clinical management and epidemiological surveillance. This study aimed to develop and diagnostically evaluate two novel species-specific loop-mediated isothermal amplification (LAMP) assays for the direct detection of D. immitis and D. repens in canine whole blood, performed in parallel in separate reactions, with emphasis on simplified and potentially near-point-of-care applicability. Primers targeting mitochondrial COI and NADH gene regions were designed and validated. In silico specificity analysis against 13 filarioid species confirmed the absence of non-specific primer binding. A direct closed-tube LAMP protocol using sodium hydroxide–Chelex-100 lysis was optimized, enabling amplification without conventional DNA extraction while reducing contamination risk and processing time to under 60 min. Relative diagnostic performance was evaluated relative to quantitative real-time PCR (qPCR) results. Using purified DNA, the D. repens assay achieved 100% relative sensitivity and relative specificity, whereas the D. immitis assay showed 94.5% relative sensitivity and 100% specificity. In direct whole-blood assays, relative specificity remained 100% for both targets, while sensitivity decreased to 90.9% for D. immitis and 77.42% for D. repens, with most false-negative reactions associated with high qPCR Ct values (>30). These findings demonstrate that the proposed assays provide a rapid and practical molecular diagnostic approach with potential applicability for point-of-care veterinary testing. Full article

(This article belongs to the Special Issue Zoonotic Parasites at the Human-Animal Interface: Advances in Epidemiology, Transmission, Diagnosis, and Treatment)

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16 pages, 18502 KB

Open AccessArticle

Morphometric Analysis of Foramina in the Middle Cranial Fossa of Dogs: A Retrospective Cone-Beam CT Study

by Nimet Turgut, Sadullah Bahar, Tutku Mecit, Yağmur Çaltıner and Abdullah Bilal Çil

Animals 2026, 16(12), 1819; https://doi.org/10.3390/ani16121819 (registering DOI) - 12 Jun 2026

Although extensively studied in humans, data on the middle cranial fossa foramina remain limited in dogs, despite their different skull morphology and high relevance to veterinary neurology, surgery and oncology. In this retrospective anatomic study, we aimed to fill this gap by presenting [...] Read more.

Although extensively studied in humans, data on the middle cranial fossa foramina remain limited in dogs, despite their different skull morphology and high relevance to veterinary neurology, surgery and oncology. In this retrospective anatomic study, we aimed to fill this gap by presenting the morphometric data of these foramina in domestic dogs of different breeds, ages, body weights, and skull sizes. The study used CBCT images of 40 dogs. Dogs were divided into three groups (small, medium, and large), regardless of sex, body weight, and breed, using neurocranium length. Then, morphological and morphometric analyses of the foramina were performed. The neurocranium length of each group differed significantly from the others (p < 0.001). In each group, the orbital fissure and round and oval foramina were bilaterally located rostrally to caudally and were of similar size (p > 0.05). While the orbital fissure was a canal in 80% of dogs, in dogs with medium and large skull sizes (17.5%), the spinous foramen showed variation, becoming both a foramen and a canal. The opening sizes increased along with the skull size (p < 0.001); the widest opening was the orbital fissure, and the narrowest opening (except for the spinous foramen) was the oval foramen. The findings may guide skull base surgeries, regional anesthesia, and the diagnosis of cranial nerve dysfunctions. Furthermore, a classification based on neurocranial length is anticipated to provide more objective craniometric measurements in animals with diverse head types and body weights. Full article

(This article belongs to the Section Companion Animals)

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22 pages, 36902 KB

Open AccessArticle

Transcriptome and 16S rRNA Amplicon Sequencing Analysis of Nutrition Metabolism in Silver Pomfret at Varying Flow Rates

by Jiabao Hu, Yuanbo Li, Youyi Zhang, Rongyue Zheng, Xiaojun Yan, Man Zhang, Yajun Wang and Lingling Jia

Animals 2026, 16(12), 1818; https://doi.org/10.3390/ani16121818 (registering DOI) - 12 Jun 2026

Silver pomfret (Pampus argenteus), a highly valued marine fish, faces challenges in aquaculture due to its sensitivity to environmental conditions. Recirculating aquaculture system (RAS) is likely to become a primary indoor cultivation method for silver pomfret in the future, so studying [...] Read more.

Silver pomfret (Pampus argenteus), a highly valued marine fish, faces challenges in aquaculture due to its sensitivity to environmental conditions. Recirculating aquaculture system (RAS) is likely to become a primary indoor cultivation method for silver pomfret in the future, so studying hydrodynamic characteristics at varying flow rates in silver pomfret RAS is crucial and has far-reaching implications for both aquaculture practices and economic returns. This study investigated the effects of water flow rates (low: 400 L/h, moderate: 600 L/h, high: 800 L/h) on the growth performance, nutritional metabolism, and gut microbiota of silver pomfret over an 8-week period. Transcriptome and 16S rRNA sequencing revealed that the moderate-flow-rate group exhibited superior growth performance, with enhanced expression of genes related to protein synthesis (HYOU1, PDIA6, ITGA11) and redox regulation (GLUL, DUSP1, GST). Additionally, the moderate flow rate promoted gut microbial diversity and stability, with higher abundances of fermentation- and chemoheterotrophy-related functions, suggesting improved nutrient metabolism. In contrast, high flow rates induced anaerobic metabolism, leading to lactate accumulation and reduced growth. These findings demonstrate that a moderate flow rate of 600 L/h optimizes silver pomfret aquaculture by enhancing growth, metabolic efficiency, and microbial health, providing a foundation for sustainable large-scale farming practices. Full article

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1 pages, 121 KB

Open AccessRetraction

RETRACTED: Masataka, N. Possible Anxiolytic Effects of Cannabidiol (CBD) Administration on Feline Responses to a Fear Response Test. Animals 2025, 15, 1642

by Nobuo Masataka

Animals 2026, 16(12), 1817; https://doi.org/10.3390/ani16121817 - 12 Jun 2026

The journal retracts the article titled “Possible Anxiolytic Effects of Cannabidiol (CBD) Administration on Feline Responses to a Fear Response Test” [...] Full article

(This article belongs to the Section Companion Animals)

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