Animals

19 pages, 11838 KB

Open AccessArticle

Transcriptomic Analysis of the Antiviral Responses in Ovine Type II Alveolar Epithelial Cells During Early Stage of Bluetongue Virus Infection

by Yunyi Chen, Nijing Lei, Zhenghao Ye, Shaohua Pu, Shimei Luo, Xianping Ma, Shaoyu Yang, Guanghua Wang, Huaijie Jia and Huashan Yi

Animals 2026, 16(2), 243; https://doi.org/10.3390/ani16020243 - 13 Jan 2026

Abstract

Bluetongue virus (BTV) infects various ruminant species, posing significant threats to animal health and causing substantial economic losses to the livestock industry. Ovine type II alveolar epithelial cells (OAECIIs) play crucial roles in maintaining pulmonary structural integrity and modulating immune responses. Their dysfunction [...] Read more.

Bluetongue virus (BTV) infects various ruminant species, posing significant threats to animal health and causing substantial economic losses to the livestock industry. Ovine type II alveolar epithelial cells (OAECIIs) play crucial roles in maintaining pulmonary structural integrity and modulating immune responses. Their dysfunction is closely associated with lung disease pathogenesis, making them important therapeutic targets. However, OAECIIs’ immunoregulatory functions and early response mechanisms during BTV infection remain unclear. To address this, we analyzed transcriptomic changes in OAECIIs following BTV-1 infection. RNA-seq revealed 1047 and 852 differentially expressed genes (DEGs) at 8 and 12 h post-infection (hpi), respectively, compared to uninfected controls. Bioinformatics analysis showed significant upregulation of nucleic acid-sensing receptors, interferon-stimulating factors, inflammatory mediators, and cytokines during early infection, mediated primarily through type I interferon signaling, TNF signaling, and cytosolic DNA-sensing pathways. We identified MAD5, ZNFX1, cGAS, OAS, PKR and ZBP1 as key pattern recognition receptors in OAECIIs during BTV infection. The IFN-β, MX1/2, RSAD2 and PLSCR1 pathways mediated antiviral responses, while IL-15, CXCL10, CCL2 triggered inflammatory responses, collectively causing structural alterations through AQP1/9 and tight junction protein modulation. These findings provide critical insights into early antiviral mechanisms and cellular structural changes in OAECIIs during BTV infection, establishing a foundation for understanding pneumonia pathogenesis and developing targeted BTV therapies. Full article

(This article belongs to the Topic Advances in Infectious and Parasitic Diseases of Animals)

► Show Figures

Figure 1

9 pages, 825 KB

Open AccessCommunication

Long-Term Immunogenicity and Protection of a rHVT-H9/Y280 Vaccine Against H9N2 Avian Influenza Virus in Commercial Layers with High Maternal Antibodies

by Sang-Won Kim, Jong-Yeol Park, Ji-Eun Son, Kai-Qiong Zheng, Cheng-Dong Yu, Ki-Woong Kim, Won-Bin Jeon, Yu-Ri Choi, Hyung-Kwan Jang, Bai Wei and Min Kang

Animals 2026, 16(2), 242; https://doi.org/10.3390/ani16020242 - 13 Jan 2026

Abstract

The endemicity of H9N2 avian influenza viruses (AIVs), particularly the Y280 lineage, poses persistent challenges to the poultry industry due to the limitations of inactivated vaccines, such as interference by maternally derived antibodies (MDAs) and incomplete suppression of viral replication. This study evaluated [...] Read more.

The endemicity of H9N2 avian influenza viruses (AIVs), particularly the Y280 lineage, poses persistent challenges to the poultry industry due to the limitations of inactivated vaccines, such as interference by maternally derived antibodies (MDAs) and incomplete suppression of viral replication. This study evaluated the immunogenicity and protective efficacy of a novel recombinant turkey herpesvirus vaccine expressing the hemagglutinin gene of H9N2/Y280 (rHVT-H9/Y280) in commercial Hy-Line Brown layers with high-MDA backgrounds. In a comparative challenge study, the rHVT-H9/Y280 vaccine induced complete protection against a homologous Y280 strain challenge at 4 weeks of age, whereas commercial inactivated vaccines failed to completely block replication, showing virus isolation rates of 16.7–25%. Long-term serological monitoring demonstrated that the rHVT-H9/Y280 vaccine elicited a robust humoral response characterized by persistent maintenance of high HI titers (>8.0 log₂) up to 39 weeks post-vaccination. These findings confirm that rHVT-H9/Y280 effectively overcomes MDA interference and provides protection by inhibition of viral replication in layer chickens, making it a promising candidate for the effective control of H9N2 AIV in endemic regions. Full article

(This article belongs to the Special Issue Epidemiology, Surveillance, and Prevention Strategies for Transboundary Animal Diseases)

► Show Figures

Figure 1

13 pages, 657 KB

Open AccessArticle

Evaluation of Pulsed Alternating Wavelength System Lighting on the Welfare Quality and Serotonin Turnover of Commercial Laying Hens Throughout a Lay Cycle

by Brittney J. Emmert, Sara Tonissen, Jenna M. Schober, Gregory S. Fraley and Darrin M. Karcher

Animals 2026, 16(2), 241; https://doi.org/10.3390/ani16020241 - 13 Jan 2026

Abstract

Laying hens require lighting for proper development and reproduction. There is limited research on the effects that lighting types have on birds’ welfare quality. A novel lighting source, Pulsed Alternating Wavelength System (PAWS), is being evaluated in the industry that claims to improve [...] Read more.

Laying hens require lighting for proper development and reproduction. There is limited research on the effects that lighting types have on birds’ welfare quality. A novel lighting source, Pulsed Alternating Wavelength System (PAWS), is being evaluated in the industry that claims to improve birds’ growth rate, decrease age at first egg, and decrease aggressive and nervous behaviors. Understanding how PAWS effects hen’s welfare, both physically and physiologically, is critical if this technology is to be adopted by industry. The project evaluated the effects of two PAWS lighting recipes on neurotransmitter turnover and welfare quality of commercial, conventionally caged laying hens. Three flocks of White leghorn hens (control [fluorescent lights] and two PAWS flocks [PAWS1 and PAWS2]) were sampled from 22 to 70 weeks of age, depending on the flock. The physical welfare of 50 hens per flock and neurotransmitter turnover of 10 hens per flock were assessed at each timepoint. The majority of welfare quality parameters were influenced by age as opposed to lighting type. No differences in dopamine turnover were observed. The hens housed under PAWS1 had reduced serotonin turnover, thus increased serotonin activity, and PAWS2 hens had improved keel bone damage scores; both indicative of improved welfare compared to control hens. The novel lighting may be beneficial to layer welfare, which may lead to increased longevity and productivity. Implementation in cage-free housing should be explored to delve into potential behavioral differences that could further influence welfare outcomes. Full article

(This article belongs to the Section Poultry)

13 pages, 4044 KB

Open AccessCase Report

A Stepwise Integrative Approach to Managing a Refractory Recurrent Cervical Sialocele in a Dog

by Suhyun Lee, Sang-Kun Jang, Duwhan Park and Hwi-Yool Kim

Animals 2026, 16(2), 240; https://doi.org/10.3390/ani16020240 - 13 Jan 2026

Abstract

Recurrent canine cervical sialocele is most often caused by incomplete excision of the mandibular–sublingual gland complex, leading to anatomical distortion and concealment of residual tissue. This case describes the multimodal management of a repeatedly recurrent cervical sialocele in a young, small-breed dog following [...] Read more.

Recurrent canine cervical sialocele is most often caused by incomplete excision of the mandibular–sublingual gland complex, leading to anatomical distortion and concealment of residual tissue. This case describes the multimodal management of a repeatedly recurrent cervical sialocele in a young, small-breed dog following multiple previous revision surgeries. A stepwise bridging strategy was adopted before definitive salvage surgery. Oral phenobarbital was instituted, resulting in partial reduction in fluid accumulation and improved comfort. Ultrasound-guided intracavitary sclerotherapy with OK-432 was then performed, inducing a localized fibrotic response but without durable cure. Final resolution was achieved only after salvage ventral paramedian (VPM) sialoadenectomy, which provided wide exposure for complete excision of deeply concealed sublingual remnant tissue within a fibrotic pseudocapsule. Histology confirmed a cervical sialocele. Transient neuropraxia resolved within 3 weeks, and no recurrence was observed at 6 months postoperatively. To the authors’ knowledge, this is the first report describing intracavitary OK-432 sclerotherapy as part of a staged multimodal strategy for canine cervical sialocele. This case illustrates the feasibility of integrating medical salivary suppression and minimally invasive sclerotherapy as bridging measures before salvage VPM surgery for refractory cervical sialocele. Full article

(This article belongs to the Section Veterinary Clinical Studies)

► Show Figures

Figure 1

28 pages, 1237 KB

Open AccessSystematic Review

Nitrogen Source–Carbohydrate Synchronization in Ruminant Nutrition: A Systematic Review

by Leilson Rocha Bezerra, Juliana Paula Felipe de Oliveira, Antônio Fernando de Melo Vaz, Kevily Henrique de Oliveira Soares de Lucena, Lucas de Souza Barros, Yuri Martins de Andrade Fortunato, Severino Gonzaga Neto, Elzania Sales Pereira, Ronaldo Lopes Oliveira and José Morais Pereira, Filho

Animals 2026, 16(2), 239; https://doi.org/10.3390/ani16020239 - 13 Jan 2026

Abstract

The synchronization between nitrogen sources and carbohydrate fractions represents a critical factor for optimizing microbial protein synthesis and overall ruminant performance. This systematic review, conducted according to PRISMA 2020 guidelines, comprehensively evaluated the interactions between different nitrogen sources (true protein, urea, controlled-release urea, [...] Read more.

The synchronization between nitrogen sources and carbohydrate fractions represents a critical factor for optimizing microbial protein synthesis and overall ruminant performance. This systematic review, conducted according to PRISMA 2020 guidelines, comprehensively evaluated the interactions between different nitrogen sources (true protein, urea, controlled-release urea, and bypass amino acids) and carbohydrate fractions (rapidly degrading soluble, slowly degrading soluble, fibrous, non-fibrous, and Van Soest fractions) in ruminant nutrition. A comprehensive search across PubMed, ScienceDirect, Web of Science, and Scopus databases identified 1855 records, of which 164 studies met the eligibility criteria for qualitative synthesis and 89 for quantitative meta-analysis. The review reveals that synchronization effectiveness varies significantly depending on the nitrogen source–carbohydrate combination, with controlled-release urea showing superior synchrony with slowly degrading carbohydrates, while conventional urea performs better with rapidly degrading sources. Meta-analytical results indicate that optimal nitrogen–carbohydrate synchronization can improve microbial protein synthesis by 18–34%, reduce urinary nitrogen excretion by 12–28%, and enhance feed efficiency by 8–15%. These findings provide evidence-based recommendations for precision nutrition strategies in ruminant production systems. Full article

(This article belongs to the Section Animal Nutrition)

10 pages, 648 KB

Open AccessCommunication

How Dairy Cows Are Culled from Freestall-Housed Dairy Herds in Wisconsin

by Kaitlin I. Buterbaugh, Thomas B. Naze and Nigel B. Cook

Animals 2026, 16(2), 238; https://doi.org/10.3390/ani16020238 - 13 Jan 2026

Abstract

Efforts to improve efficiency and profitability on dairy farms have renewed focus on how culling practices affect herd sustainability and economic outcomes. This study surveyed decision-makers on 60 high-producing, freestall-housed dairy farms in Wisconsin, with a mean (SD) turnover rate of 36.0 (8.0)%. [...] Read more.

Efforts to improve efficiency and profitability on dairy farms have renewed focus on how culling practices affect herd sustainability and economic outcomes. This study surveyed decision-makers on 60 high-producing, freestall-housed dairy farms in Wisconsin, with a mean (SD) turnover rate of 36.0 (8.0)%. Using a structured questionnaire, we examined herd management, culling criteria, and motivations. Most farms (93%) used on-farm management systems to guide culling, yet only 48% used designated reports, relying instead on individual cow records. Milk production, infertility, and somatic cell count were the top culling criteria, with high milk yield cited as the most difficult factor in removal decisions. While 54% recorded the most obvious reason for culling, only 7% documented multiple causes. Cull cows were typically transported by third parties; 80% farms sent cows directly to slaughter, while 52% sent them to auction. One-third of farms sold cows for continued dairy use. Euthanasia was performed on 93% of farms, mostly by employees, with minimal veterinary input. The study aimed to investigate producer perspectives on the culling decision-making process on commercial dairy farms. The findings highlight opportunities for improved veterinary involvement and the use of structured herd-level reports to support more strategic culling decisions. Full article

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

► Show Figures

Figure 1

30 pages, 8192 KB

Open AccessArticle

Structural Insights into the Receptor-Binding Domain of Bat Coronavirus HKU5-CoV-2: Implications for Zoonotic Transmission via ACE2

by Manal A. Babaker, Nariman Sindi, Othman Yahya Alyahyawy, Ehssan Moglad, Mohieldin Elsayid, Thamir M. Eid, Mohamed Eltaib Elmobark and Hisham N. Altayb

Animals 2026, 16(2), 237; https://doi.org/10.3390/ani16020237 - 13 Jan 2026

Abstract

The zoonotic potential of bat coronaviruses, especially HKU5, is a significant issue because of their capacity to utilize human angiotensin-converting enzyme 2 (ACE2) as a receptor for cellular entry. This study offers structural insights into the binding kinetics of HKU5 (Bat Merbecovirus HKU5) [...] Read more.

The zoonotic potential of bat coronaviruses, especially HKU5, is a significant issue because of their capacity to utilize human angiotensin-converting enzyme 2 (ACE2) as a receptor for cellular entry. This study offers structural insights into the binding kinetics of HKU5 (Bat Merbecovirus HKU5) receptor-binding domain (RBD) spike protein with human ACE2 through a multiscale computational method. This study employed structural modeling, 300-nanosecond (ns) molecular dynamics (MD) simulations, alanine-scanning mutagenesis, and computational peptide design to investigate ACE2 recognition by the HKU5 RBD and its interactions with peptides. The root mean square deviation (RMSD) investigation of HKU5–ACE2 complexes indicated that HKU5 exhibited greater flexibility than SARS-CoV-2, with RMSD values reaching a maximum of 1.2 nm. Free energy analysis, Molecular Mechanics/Generalized Born Surface Area (MM/GBSA), indicated a more robust binding affinity of HKU5 to ACE2 (ΔG_Total = −21.61 kcal/mol) in contrast to SARS-CoV-2 (ΔG_Total = −5.82 kcal/mol), implying that HKU5 binding with ACE2 had higher efficiency. Additionally, a peptide was designed from the ACE2 interface, resulting in the development of 380 single-site mutants by mutational alterations. The four most promising mutant peptides were selected for 300-nanosecond (ns) MD simulations, subsequently undergoing quantum chemical calculations (DFT) to evaluate their electronic characteristics. MM/GBSA of −37.83 kcal/mol indicated that mutant-1 exhibits the most favorable binding with HKU5, hence potentially inhibiting ACE2 interaction. Mutant-1 formed hydrogen bonds involving Glu74, Ser202, Ser204, and Asn152 residues of HKU5. Finally, QM/MM calculations on the peptide–HKU5 complexes showed the most favorable ΔE_interaction of −170.47 (Hartree) for mutant-1 peptide. These findings offer a thorough comprehension of receptor-binding dynamics and are crucial for evaluating the zoonotic risk associated with HKU5-CoV and guiding the design of receptor-targeted antiviral treatments. Full article

(This article belongs to the Special Issue Zoonotic Diseases: Etiology, Diagnosis, Surveillance and Epidemiology: Second Edition)

► Show Figures

Figure 1

26 pages, 2421 KB

Open AccessReview

Fatty Liver in Fish: Metabolic Drivers, Molecular Pathways and Physiological Solutions

by Xiyu Xie, Chaoyang Zhang, Ilham Zulfahmi, Esau Mbokane and Quanquan Cao

Animals 2026, 16(2), 236; https://doi.org/10.3390/ani16020236 - 13 Jan 2026

Abstract

Fatty liver in fish is characterized by excessive lipid accumulation, driven by factors such as inflammation, oxidative stress, and the overexpression of lipid-related genes. This condition can lead to metabolic dysfunction and reduced disease resistance, resulting in growth disorders and even mortality. Increasing [...] Read more.

Fatty liver in fish is characterized by excessive lipid accumulation, driven by factors such as inflammation, oxidative stress, and the overexpression of lipid-related genes. This condition can lead to metabolic dysfunction and reduced disease resistance, resulting in growth disorders and even mortality. Increasing incidence of fatty liver is closely linked to environmental conditions and feeding practices, posing significant challenges to the aquaculture industry. This paper offers a comprehensive overview of hepatic steatosis, with a particular emphasis on fish species. Through a detailed review of various scholarly works, this paper seeks to identify common patterns, emerging trends, and measurable correlations, highlighting the critical importance of understanding this complex relationship. The study of fatty liver is conducted across three dimensions: influencing factors, underlying mechanisms, and potential solutions. Currently, numerous factors contribute to the development of fatty liver, such as feed composition and environmental temperature. On a mechanistic level, the research explores lipid accumulation, inflammation, oxidative stress, and related processes. Furthermore, the paper suggests various solutions and preventive strategies, including considering environmental adaptability during animal migration, employing genetic enhancement techniques, modifying feeding practices, investigating the Nrf2 pathway, and utilizing rapamycin. These findings have significant implications for fisheries management and aquaculture practices, providing valuable insights to enhance sustainability in the industry. Full article

(This article belongs to the Special Issue Developments in Environmental Physiology, Nutrition Physiology, and Immunity Mechanisms of Aquatic Animals)

► Show Figures

Figure 1

14 pages, 760 KB

Open AccessArticle

Two-Generation Genetic Evaluation of Female Reproductive Performance in Pacific White Shrimp (Penaeus vannamei) Under SPF Conditions

by Jiaqi Yu, Jie Kong, Sheng Luan, Jiawang Cao, Mianyu Liu, Kun Luo, Jian Tan, Ping Dai, Zhaoxin Wang, Juan Sui and Xianhong Meng

Animals 2026, 16(2), 235; https://doi.org/10.3390/ani16020235 - 13 Jan 2026

Abstract

Reproductive inefficiency remains a major constraint in Penaeus vannamei hatcheries due to high rates of non-spawning females. This study presents the first two-generational quantitative genetic analysis of female reproductive performance under standardized SPF (Specific Pathogen-Free) conditions. A total of 986 females across two [...] Read more.

Reproductive inefficiency remains a major constraint in Penaeus vannamei hatcheries due to high rates of non-spawning females. This study presents the first two-generational quantitative genetic analysis of female reproductive performance under standardized SPF (Specific Pathogen-Free) conditions. A total of 986 females across two generations (2021–2022) from 198 full-sib and 68 half-sib families were evaluated. Traits analyzed included spawning frequency (SF), mean spawning interval (MSI), number of eggs laid for the first time (NE1), average spawning (AS), total spawning (TS), and spawning success (SS). Heritability estimates for SF, SS, and TS were moderate (0.30 ± 0.06, 0.23 ± 0.06 and 0.28 ± 0.07, respectively), while MSI, NE1, and AS showed low heritability (0.10–0.16). When analyzed separately by year, heritability estimates declined substantially for most traits in the second generation. Strong positive genetic correlations were observed between SF, MSI, NE1, AS, and TS, with pairwise estimates ranging from 0.82 to 0.99, indicating that these traits are under shared genetic control. Despite not being direct selection objects, all reproductive traits exhibited relative genetic progress (246–488% per generation), which is attributable to the high selection intensity applied to the parental generation. Our findings provide a robust foundation for integrating reproductive performance into breeding programs for P. vannamei, particularly under biosecurity and commercial feed-dominated conditions. Full article

(This article belongs to the Section Aquatic Animals)

► Show Figures

Figure 1

24 pages, 1256 KB

Open AccessArticle

Non-Invasive Assessment of Adrenal Activity in the Subterranean Rodent Ctenomys talarum in Field and Laboratory Conditions

by Roxana Zenuto, Valentina Brachetta, María Celina Carrizo, María Sol Fanjul and Cristian Eric Schleich

Animals 2026, 16(2), 234; https://doi.org/10.3390/ani16020234 - 13 Jan 2026

Abstract

The endocrine stress response is a valuable tool for evaluating how organisms cope with environmental challenges. However, selecting an appropriate matrix for measuring glucocorticoids (GCs) requires careful consideration of sample quality and accessibility. This study reveals that blood sampling affects plasma cortisol levels [...] Read more.

The endocrine stress response is a valuable tool for evaluating how organisms cope with environmental challenges. However, selecting an appropriate matrix for measuring glucocorticoids (GCs) requires careful consideration of sample quality and accessibility. This study reveals that blood sampling affects plasma cortisol levels in the subterranean rodent Ctenomys talarum, with the effect being reversed shortly thereafter. To facilitate a non-invasive approach, an enzyme-linked immunosorbent assay (EIA) that had previously been validated for measuring plasma cortisol in C. talarum was evaluated to measure adrenocortical activity by analyzing fecal glucocorticoid metabolites (FGCs). Using this assay, we monitored the stress response during wild capture, transport to captivity, adrenocorticotropic hormone (ACTH) stimulation, and immobilization. This showed that FGC levels accurately reflect adrenal activation in these contexts. We also documented a relationship between reproductive seasonality and FGCs. Finally, we provide evidence for a relationship between adrenal activity and behavior. Our results suggest that when considering plasma GCs for the assessment of acute stress, it is crucial to understand the magnitude and timing of the effects of blood sampling on the stress state of organisms. The validation of FGC measurement in C. talarum provides a new option for advancing ecophysiological studies in both the wild and captivity. Full article

(This article belongs to the Special Issue Wild Animal Welfare: Science, Ethics and Law)

► Show Figures

Figure 1

17 pages, 557 KB

Open AccessArticle

Effects of Dietary Salt and Boric Acid on Milk Quality in Savak Akkaraman Sheep

by Pelin Beyazgül, Selçukhan Akarsu, Yasin Baykalir and Ülkü Gülcihan Şimşek

Animals 2026, 16(2), 233; https://doi.org/10.3390/ani16020233 - 13 Jan 2026

Abstract

This study evaluated the effects of varying dietary salt and boric acid addition doses on the milk quality of Savak Akkaraman sheep. A total of 120 animals were as-signed to six treatment groups (n = 20): control (C), rock salt (S; 10 [...] Read more.

This study evaluated the effects of varying dietary salt and boric acid addition doses on the milk quality of Savak Akkaraman sheep. A total of 120 animals were as-signed to six treatment groups (n = 20): control (C), rock salt (S; 10 g/day), boric acid 20 mg/day (B20), boric acid 40 mg/day (B40), BS20 (20 mg boric acid + 10 g/day rock salt), and BS40 (40 mg boric acid + 10 g/day rock salt). All analyses were performed in duplicate on six samples, taken on days 30 and 35 following the administration of the additives. Physicochemical analyses only showed significant variation in milk pH (p = 0.006), while acidity, dry matter, and ash remained unaffected. Strong positive correlations were found among protein, lactose, salt, and density (r > 0.95; p < 0.001). Coagulation times differed widely, with the longest being observed in BS20 (995.03 s) and the shortest in BS40 (141.73 s). Among mineral parameters, only selenium levels differed significantly between the treatment groups (p < 0.05). No significant differences were found for fat, solids-not-fat, lactose, freezing point, or electrical conductivity. Importantly, boron addition had a significant influence on total casein content (p < 0.001). Overall, dietary rock salt and boric acid did not markedly alter the basic milk composition but produced notable physicochemical changes, particularly in coagulation behavior and casein levels, which may influence the technological properties of sheep milk. Full article

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

► Show Figures

Figure 1

26 pages, 385 KB

Open AccessReview

Mapping the Kinase Inhibitor Landscape in Canine Mammary Carcinoma: Current Status and Future Opportunities

by Małgorzata Chmielewska-Krzesińska

Animals 2026, 16(2), 232; https://doi.org/10.3390/ani16020232 - 13 Jan 2026

Abstract

Background: Canine mammary carcinoma (CMC) is the most common malignant tumour in female dogs and, due to its similarities, is a valuable comparative model for human breast cancer. Kinase inhibitors have revolutionised the treatment of human breast cancer; their use in veterinary [...] Read more.

Background: Canine mammary carcinoma (CMC) is the most common malignant tumour in female dogs and, due to its similarities, is a valuable comparative model for human breast cancer. Kinase inhibitors have revolutionised the treatment of human breast cancer; their use in veterinary oncology remains marginal. Aim: This review summarises the current knowledge of kinase signalling pathways in CMC and assesses which kinase inhibitors approved for human use have potential in veterinary medicine. Methods: A systematic search of the PubMed database from 1985 to 2025 was performed, focusing on kinase-targeted therapies in both human and canine mammary carcinomas. Data were categorised according to molecular target, clinical approval status, and available preclinical or clinical veterinary evidence. Results: Key molecular pathways targeted by kinase inhibitors are conserved across species, supporting translational opportunities. In vitro studies demonstrate that palbociclib, alpelisib, everolimus, and lapatinib inhibit growth and signalling in CMC cell lines. Clinical trials have not been conducted. Conclusions: Approved kinase inhibitors for human use have untapped therapeutic potential in veterinary oncology. Translational research, including xenograft and organoid models, followed by clinical trials in dogs, is required. Gaining this knowledge could lead to targeted treatment for dogs while advancing comparative understanding of mammary cancer biology across species. Full article

(This article belongs to the Special Issue Tumors in Companion Animals: Detection, Diagnosis, Prognosis and Treatment)

► Show Figures

Graphical abstract

26 pages, 1406 KB

Open AccessArticle

The Welfare Impact of Heat Stress in South American Beef Cattle and the Cost-Effectiveness of Shade Provision

by Cynthia Schuck-Paim, Wladimir Jimenez Alonso, Anielly de Paula Freitas, Camila Pereira de Oliveira, Vinicius de França Carvalho Fonseca and Tâmara Duarte Borges

Animals 2026, 16(2), 231; https://doi.org/10.3390/ani16020231 - 13 Jan 2026

Abstract

Heat stress represents a pervasive welfare challenge for beef cattle and other species in tropical and subtropical regions. While its physiological and production impacts are well-documented, quantitative measures of the welfare impact of heat stress remain absent. This study provides the first quantification [...] Read more.

Heat stress represents a pervasive welfare challenge for beef cattle and other species in tropical and subtropical regions. While its physiological and production impacts are well-documented, quantitative measures of the welfare impact of heat stress remain absent. This study provides the first quantification of the welfare impact of heat stress in beef cattle (mostly Nelore), estimated as cumulative time in thermal discomfort of four intensities (Annoying, Hurtful, Disabling, Excruciating) using the Welfare Footprint Framework. We analyzed climate data from 636 locations over five years across major beef production areas in Brazil, Argentina, Colombia, Paraguay, and Uruguay. Daily heat stress episodes and chronic heat stress exposure were assessed, respectively, using Comprehensive Climate Index (CCI) levels and the Annual Thermal Load metric, which sums daily excesses above a threshold of thermal comfort (CCI = 30 °C) throughout the year, classifying locations into five risk categories. Welfare impacts were estimated for thirteen heat stress scenarios modeled by considering each CCI level within each thermal risk category. Beef cattle in moderate-risk regions were estimated to experience primarily mild thermal discomfort for an average of 5 h daily. This duration increased to an average of 7 h daily in high-risk areas, of which 4.5 h in moderate to intense thermal discomfort (Hurtful or higher). Very high-risk regions reached 10 h of daily thermal discomfort, while extreme-risk regions showed beef cattle facing heat stress for over 11 h on 307 days annually, including over 3 h per day under severe thermoregulatory effort. Overall, 65% of animals were in regions of high thermal risk or above, experiencing between 280 and 2800 h annually in moderate to intense thermal discomfort—a magnitude that places heat stress among the most significant welfare challenges in animal production. Shade provision reduced time in severe discomfort of Disabling intensity by 85% (from 578 to 83 h annually), with economic returns of US$12–16 per animal and payback periods of approximately 16 months. By quantifying welfare impacts as cumulative time in thermal discomfort, shade provision emerges as one of the most effective welfare interventions available for beef cattle, and likely other grazing ruminants, in tropical and subtropical regions. Full article

(This article belongs to the Section Animal Welfare)

► Show Figures

Figure 1

18 pages, 758 KB

Open AccessArticle

Effects of Sodium Butyrate and Organic Zinc Supplementation on Performance, Mineral Metabolism, and Intestinal Health of Dairy Calves

by Mellory M. Martins, Larissa S. Gheller, Bruna L. de Noronha, Gabrielly A. Cassiano, Mariana B. Figueiredo, Caroline M. Meira, Flávia F. Simili, Márcia S. V. Salles and Arlindo Saran Netto

Animals 2026, 16(2), 230; https://doi.org/10.3390/ani16020230 - 13 Jan 2026

Abstract

The objective of this study was to evaluate the effects of sodium butyrate and organic zinc supplementation, alone or combined, on performance, zinc metabolism, blood parameters, and gut health in Holstein calves highly challenged by heat and diarrhea during the pre-weaning and weaning [...] Read more.

The objective of this study was to evaluate the effects of sodium butyrate and organic zinc supplementation, alone or combined, on performance, zinc metabolism, blood parameters, and gut health in Holstein calves highly challenged by heat and diarrhea during the pre-weaning and weaning periods. Forty-eight male calves were assigned to one of four treatments: control (CON), SB (3 g/kg of sodium butyrate in dry matter [DM]), OZn (262 mg/kg of organic zinc in DM), or SBOZn (3 g/kg of sodium butyrate and 262 mg/kg of organic zinc in DM). Calves were monitored from days 7 to 63 for feed intake, weight gain, body morphometry, fecal score, and blood parameters. Zinc balance was evaluated from days 45 to 49, and 24 calves were slaughtered on day 64 for jejunal sampling to assess tight junction gene expression. Diarrhea incidence was high (>90%) across groups. Fecal scores varied over time but did not differ between treatments. The OZn and SBOZn groups had higher Zn intake, with greater absorption and retention of the mineral compared to the CON and SB groups. Additionally, the OZn group tended to have higher serum Zn concentrations. SB and OZn, separately or combined, had limited effects and did not consistently improve the performance or health of calves highly challenged during pre-weaning and weaning. Full article

(This article belongs to the Collection Feeding Strategies to Improve Sustainability and Welfare in Animal Production)

► Show Figures

Figure 1

25 pages, 20500 KB

Open AccessArticle

Early-Onset Negative Energy Balance in Transition Dairy Cows Increases the Incidence of Retained Fetal Membranes

by Zhihong Zhang, Shanshan Guo, Jianhao Yang, Xinfeng Hou, Xia Zhang, Huifeng Liu, Tao Liu and Yaping Jin

Animals 2026, 16(2), 229; https://doi.org/10.3390/ani16020229 - 13 Jan 2026

Abstract

This study investigated the metabolic mechanisms driving physiological functional remodeling in RFM by analyzing plasma biochemical parameters and metabolomic profiles at key peripartum timepoints (21 and 7 d prepartum and 4 h postpartum), integrated with placental and fetal membrane metabolic characteristics. The results [...] Read more.

This study investigated the metabolic mechanisms driving physiological functional remodeling in RFM by analyzing plasma biochemical parameters and metabolomic profiles at key peripartum timepoints (21 and 7 d prepartum and 4 h postpartum), integrated with placental and fetal membrane metabolic characteristics. The results revealed that RFM cows exhibited significant negative energy balance (NEB) as early as 21 days before parturition, characterized by elevated plasma levels of non-esterified fatty acids, β-hydroxybutyrate, and malondialdehyde, alongside reduced activity of antioxidant enzymes (GSH-Px, CAT) (p ≤ 0.05). Metabolomic analysis demonstrated persistent lipid metabolism dysregulation, amino acid imbalance, and nucleotide metabolism disturbances in RFM cows from 21 days prepartum to 4 h postpartum, indicating premature mobilization of adipose and muscle tissues. Further metabolomic analyses of the placenta and fetal membranes confirmed that metabolic dysfunction compromises energy supply during parturition, adversely affecting immune homeostasis and extracellular matrix degradation in the placenta and fetal membranes of RFM dairy cows. These physiological dysfunctions have the potential to impede the timely expulsion of fetal membranes after calving. In conclusion, RFM is closely associated with early-onset metabolic dysfunction during the periparturient period, where insufficient energy supply due to NEB, oxidative stress, and immune-endocrine disruptions collectively impair normal fetal membrane detachment. Full article

(This article belongs to the Collection Cattle Diseases)

► Show Figures

Figure 1

17 pages, 348 KB

Open AccessReview

Challenges and Methodologies to Assess Protein Requirement and Quality Across Different Life Stages in Dogs: A Review

by Lucas Bassi Scarpim and Leticia Graziele Pacheco

Animals 2026, 16(2), 228; https://doi.org/10.3390/ani16020228 - 13 Jan 2026

Abstract

Determining protein requirements (PRs) for dogs remains a longstanding challenge. During growth, the rapid rate of protein deposition increases the demand for amino acids. In adult dogs, differences in overall diet digestibility and lower energy requirements of domestic dogs have led to discrepancies [...] Read more.

Determining protein requirements (PRs) for dogs remains a longstanding challenge. During growth, the rapid rate of protein deposition increases the demand for amino acids. In adult dogs, differences in overall diet digestibility and lower energy requirements of domestic dogs have led to discrepancies between the minimum crude protein (CP) value proposed by the National Research Council (NRC; 80 g of CP/kg of diet) and the 180 g of CP/kg of diet proposed by the European Pet Food Industry Federation (FEDIAF) and the Association of American Feed Control Officials (AAFCO), although most commercially available adult dog feeds offer protein levels that exceed both recommendations. In elderly dogs, physiological changes such as sarcopenia and reduced energy intake indicate a potential increase in PR, although evidence remains scarce. A similar gap exists for pregnant and lactating bitches, since most recommendations rely on extrapolations from growth studies. Classical PR recommendations were based on body weight gain and nitrogen balance (NB), methods that present important limitations. Due to this, stable isotope methods—including ¹³C-leucine, ¹⁵N-glycine, and ¹³C-phenylalanine—have emerged as precise methodological tools, enabling a detailed and dynamic assessment of whole-body protein metabolism, protein quality, and more accurate determination of PR and recommended allowance across different life stages. Full article

(This article belongs to the Section Animal Nutrition)

14 pages, 1282 KB

Open AccessArticle

Testing the Island Effect in a Highly Mobile Pollinator: Wing Morphological Divergence in Euglossa mixta from Continental and Insular Panama

by Yostin Añino, Jordan Hernández-Martelo, Fernando Moya, Alejandro Piñeiro-González, Laura M. Pérez, Dumas Gálvez, Yosiat Vega-Rovira, Julio Trujillo, Anette Garrido, Danilo Arrocha, Franco Cruz-Jofré and Hugo A. Benítez

Animals 2026, 16(2), 227; https://doi.org/10.3390/ani16020227 - 12 Jan 2026

Abstract

Islands provide valuable opportunities to study how isolation affects phenotypic variation. Even though orchid bees are highly mobile, their movement can still be restricted by marine barriers. In this study, we assessed whether insular isolation impacts wing shape in the orchid bee Euglossa [...] Read more.

Islands provide valuable opportunities to study how isolation affects phenotypic variation. Even though orchid bees are highly mobile, their movement can still be restricted by marine barriers. In this study, we assessed whether insular isolation impacts wing shape in the orchid bee Euglossa mixta across the Coiba archipelago and a nearby mainland site in Western Panama. Our study analyzed 271 individuals using geometric morphometrics, focusing on forewing venation landmarks, and evaluated the variation using multivariate analyses of shape variation and quantifying the shape of Mahalanobis distances. Additionally, we conducted a Mantel test to explore the relationship between geographic distance and morphological divergence. Our findings reveal that wing shape variation in E. mixta is largely conserved but shows fine-scale structuring consistent with spatial patterns expected in insular systems. These results suggest that even highly mobile pollinators may experience enough isolation for subtle phenotypic shifts to occur, highlighting the sensitivity of geometric morphometrics for detecting early stages of morphological differentiation. Full article

► Show Figures

Figure 1

14 pages, 1237 KB

Open AccessArticle

Galba truncatula: Distribution, Presence in Fountains and Identification of Factors Related to Its Occurrence in Bulgaria

by Katya Georgieva and Boyko Neov

Animals 2026, 16(2), 226; https://doi.org/10.3390/ani16020226 - 12 Jan 2026

Abstract

Galba truncatula acts as an intermediate host for several parasitic flukes of veterinary importance, but a targeted study on its spatial presence as well as the impact of environmental factors in Southeastern Europe has not been conducted. During the summer months of 2017 [...] Read more.

Galba truncatula acts as an intermediate host for several parasitic flukes of veterinary importance, but a targeted study on its spatial presence as well as the impact of environmental factors in Southeastern Europe has not been conducted. During the summer months of 2017 and 2018, a survey of 191 water bodies in 14 districts in Central, Southern and Western Bulgaria was conducted, with a focus on animal drinking fountains. Each site was assessed for snail presence and characterized by altitude, temperature, precipitation, shade and type of water body. Logistic regression modeling was used to identify the important factors related to the occurrence of snail species. The frequency of habitats found was 29.3%, with no differences observed between the studied districts (p > 0.05). Snails were present across a wide range of altitudes (78–1926 m), annual mean temperature (7.8–14.0 °C) and annual mean precipitation (523–796 mm). The high habitat frequencies were recorded in streams (60.0%) and on the banks on small rivers (50.0%). The presence of snails in the two studied types of fountains (without or with a concrete platform) was 24.1% and 17.2%, respectively, with no significant difference between them (p > 0.05). Regression analysis revealed temperature, shade, and type of water body as factors that could significantly influence the spatial presence of G. truncatula. The findings demonstrate the ecological adaptability of G. truncatula and highlight its presence in habitats with high potential for contact with domestic and wild ruminants. This information fills a regional knowledge gap and can support risk assessment and control measures for fluke-borne diseases in livestock and wildlife. Full article

(This article belongs to the Section Wildlife)

20 pages, 4492 KB

Open AccessArticle

Integrated Analysis of Testicular Histology, Sperm Quality, and Gene Expression (TGFB2, DMRT1) in Rooster Semen (Gallus gallus domesticus)

by Anastasiya Ivershina, Yuliya Silyukova, Elena Fedorova, Olga Stanishevskaya, Irina Mirzakaeva and Marina Pozovnikova

Animals 2026, 16(2), 225; https://doi.org/10.3390/ani16020225 - 12 Jan 2026

Abstract

The study of the relationship between testicular morphology and sperm quality is a pressing issue, for which molecular genetic approaches, including quantitative analysis of gene expression, are being implemented. The aim of this study was to identify correlations between the histomorphological structure of [...] Read more.

The study of the relationship between testicular morphology and sperm quality is a pressing issue, for which molecular genetic approaches, including quantitative analysis of gene expression, are being implemented. The aim of this study was to identify correlations between the histomorphological structure of the testes, fresh sperm parameters, and the expression level of key spermatogenesis genes—TGFB2 and DMRT1—in roosters. The experiment was conducted on 10 Russian Snow White roosters aged 28–32 weeks. Sperm quality was assessed by volume, sperm concentration, total and progressive motility, and viability; histological analysis of the rooster testes was performed. The relative expression of the TGFB2 and DMRT1 genes in sperm was analyzed. Multiple correlation analysis of the data was conducted. A positive correlation was found between ejaculate volume and the number of spermatogonia (p = +0.651), a negative correlation between ejaculate volume and the number of second-order spermatocytes (p = −0.704), a negative correlation between the total cross-sectional area of the seminiferous tubules of the testes and sperm viability (p = −0.782), a negative correlation between the number of seminiferous tubules and the average diameter of their cross-section (p = −0.685), and a positive correlation between total and progressive sperm motility (p = +0.794). Analysis of TGFB2 and DMRT1 gene expression in sperm demonstrated a certain relationship between molecular genetic mechanisms and histomorphometric parameters. The expression level of the DMRT1 gene, which plays a key role in sex determination in birds during embryogenesis, had a number of negative correlations with such parameters as testicle weight (r = −0.782), total/progressive sperm motility (r = −0.552; r = −0.612), and viability (r = −0.552). Expression of the TGFB2 gene had no significant relationship with the studied parameters, but correlation analysis revealed a moderate positive relationship (r = +0.321) with DMRT1 gene expression. The data obtained indicate the expediency of integrating morphometric, cellular, and molecular analysis for an objective assessment of rooster reproductive function. Full article

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

► Show Figures

Figure 1

Journal Description

Animals

Latest Articles

Journal Menu

Journal Browser

Highly Accessed Articles

Latest Books

E-Mail Alert

News

Topics

Conferences

Special Issues

Topical Collections

Further Information

Guidelines

MDPI Initiatives

Follow MDPI