Animals

13 pages, 3670 KiB

Open AccessArticle

Diagnostic Approach and Pathological Characterization of Metastatic Intrahepatic Cholangiocarcinoma in a Captive Puma (Puma concolor)

by Elisa Mazzotta, Claudia Zanardello, Giovanni De Zottis, Antonio Barberio, Mery Campalto, Federico Martignago, Giulia Maria De Benedictis, Carlo Guglielmini, Francesca Zanusso and Greta Foiani

Animals 2025, 15(12), 1821; https://doi.org/10.3390/ani15121821 - 19 Jun 2025

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Abstract

A 16-year-old captive male puma (Puma concolor) presented with progressive weight loss and acute mild gastrointestinal symptoms. Clinical examination, and diagnostic imaging revealed abdominal, pleural, and pericardial effusion and nodular masses in multiple organs. Due to the clinical condition, multiple organ [...] Read more.

A 16-year-old captive male puma (Puma concolor) presented with progressive weight loss and acute mild gastrointestinal symptoms. Clinical examination, and diagnostic imaging revealed abdominal, pleural, and pericardial effusion and nodular masses in multiple organs. Due to the clinical condition, multiple organ involvement, and a poor prognosis, compassionate euthanasia was chosen. Necropsy revealed abundant cavitary effusions and multiple yellow–white neoplastic masses involving the left hepatic lobes. Metastatic nodules were observed throughout the peritoneum, pericardium, kidney, spleen, and spermatic cord. Histologically, the tumor was characterized by tubular and acinar structures embedded in a desmoplastic stroma. Neoplastic cells were positive for pan-cytokeratins (CKs), CK7, and, to a lesser extent, CK20. The gross, histologic, and immunohistochemical findings were consistent with intrahepatic metastatic cholangiocarcinoma. Full article

(This article belongs to the Special Issue Wildlife Diseases: Pathology and Diagnostic Investigation)

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12 pages, 679 KiB

Open AccessArticle

A Novel Echocardiographic Index (Modified-Left-Atrium-to-Aorta Ratio) for Quantifying Left Atrial Size and Differentiating Stages in Dogs with Myxomatous Mitral Valve Disease

by Minsuk Kim, Minwoong Seo and Chul Park

Animals 2025, 15(12), 1820; https://doi.org/10.3390/ani15121820 - 19 Jun 2025

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Abstract

Myxomatous mitral valve disease (MMVD) is the most common heart disease in small-breed dogs, and accurate assessment of the left atrial (LA) size is essential for diagnosis and management. The traditional echocardiographic method, LA-to-Aorta (LA/Ao) ratio, is widely used, but evaluates LA size [...] Read more.

Myxomatous mitral valve disease (MMVD) is the most common heart disease in small-breed dogs, and accurate assessment of the left atrial (LA) size is essential for diagnosis and management. The traditional echocardiographic method, LA-to-Aorta (LA/Ao) ratio, is widely used, but evaluates LA size in only one view. This study introduces a novel index—the modified-LA/Ao (M-LA/Ao) ratio—which combines two echocardiographic views to better reflect the true LA size. This study retrospectively analyzed thoracic radiographs and echocardiograms from 136 dogs, including healthy controls and dogs with MMVD classified into stages B1, B2, and C according to ACVIM guidelines. The ability of the M-LA/Ao ratio is compared to conventional indices using correlation analysis and receiver operating characteristic (ROC) curves. The M-LA/Ao ratio showed a strong correlation with existing indices and slightly improved discrimination between ACVIM stages B1 and B2, although performance between stages B2 and C was similar to the LA/Ao ratio. Intra- and interobserver variability were also acceptable. Our findings suggest that the M-LA/Ao ratio may provide a practical and sensitive method to evaluate LA enlargement in early-stage MMVD, helping clinicians detect subtle cardiac remodeling before progression to advanced disease. Full article

(This article belongs to the Section Veterinary Clinical Studies)

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21 pages, 4535 KiB

Open AccessArticle

Effects of Biological Characteristics and Environmental Factors on Swimming Performance of Endemic Fish in Southwest China

by Jianing Rao, Zhiguang Zhang, Yuanming Wang, Qi Wei, Guoqing Chen, Xintong Li, Ruifeng Liang and Kefeng Li

Animals 2025, 15(12), 1819; https://doi.org/10.3390/ani15121819 - 19 Jun 2025

Viewed by 158

Abstract

Swimming is crucial to fish survival and reproduction, and is usually influenced by species, morphology, habitat environment and physiological condition. Understanding the changes in fish swimming ability and identifying the driving factors behind these changes is vital. In this study, seven endemic fish [...] Read more.

Swimming is crucial to fish survival and reproduction, and is usually influenced by species, morphology, habitat environment and physiological condition. Understanding the changes in fish swimming ability and identifying the driving factors behind these changes is vital. In this study, seven endemic fish species from the Qingshui River, an important tributary of the upper Pearl River in Southwest China, were used to explore the effects of biological and environmental factors on fish swimming ability. The results indicated that the body length parameters had a minimal effect on the induced swimming speed (U_ind) but played a positive role in critical and burst swimming ability (U_crit and U_burst). Weight (W) and W/SL (the ratio of weight to standard body length) correlated more highly with swimming ability than body length parameters. Fish preferring the lotic environment exhibited higher U_crit and U_burst, and fish with a streamlined morphology had a smaller U_ind. The U_ind, U_ind/SL, U_ind/W and U_ind/(W/SL) of Discogobio yunnanensis (Regan, 1907) and Pseudocrossocheilus tridentis (Cui & Chu, 1986) were significantly higher during the spawning period. Acrossocheilus yunnanensis (Regan, 1904) performed better during the spawning period in U_crit and U_crit/SL. The U_burst of fish during the spawning period was smaller than that during the non-spawning period. Generally, the species difference had the greatest contribution to the swimming ability difference, followed by environmental conditions and fish morphology. Full article

(This article belongs to the Section Aquatic Animals)

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10 pages, 345 KiB

Open AccessFeature PaperArticle

Alcoholization as an Alternative Treatment for Prostatic Cyst and Abscess in Dogs

by Giulia Ballotta, Giuseppe Spinella, Marco Cunto and Daniele Zambelli

Animals 2025, 15(12), 1818; https://doi.org/10.3390/ani15121818 - 19 Jun 2025

Viewed by 172

Abstract

Current therapeutic approaches for prostatic cysts and abscesses involve surgical methods like reduction and omentalization. This retrospective study evaluates the effectiveness and safety of ultrasound-guided percutaneous drainage combined with alcoholization as a minimally invasive treatment for prostatic cysts and abscesses in dogs. Forty-three [...] Read more.

Current therapeutic approaches for prostatic cysts and abscesses involve surgical methods like reduction and omentalization. This retrospective study evaluates the effectiveness and safety of ultrasound-guided percutaneous drainage combined with alcoholization as a minimally invasive treatment for prostatic cysts and abscesses in dogs. Forty-three male dogs with a single prostatic cavity (1–6 cm) were enrolled and categorized into two groups: group A (twenty-two dogs with prostatic abscesses treated via alcoholization) and group B (twenty-one dogs with BPH-associated cysts). Group B was further subdivided into three subgroups: B1 (finasteride only), B2 (alcoholization only), and B3 (combined finasteride and alcoholization). Alcoholization involved sedation, drainage of the cavity, and two ethanol instillations (first: 1/3–1/4 of aspirated volume; second: 1/10 after 25 min). Ultrasound evaluations were performed at scheduled intervals of up to 150 days. In group A, 76.19% of abscesses were resolved within 30 days, with complete resolution in all dogs by day 150. In group B, the combined treatment (group B3) showed superior outcomes, with significant cyst diameter reduction observed in all dogs within 30 days and complete resolution in 62.5% by day 150. These findings support alcoholization as an effective, low-complication alternative to surgery for selected prostatic lesions in dogs. Full article

(This article belongs to the Section Veterinary Clinical Studies)

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17 pages, 1701 KiB

Open AccessArticle

The Effects of the Lactation Period, Mare Age, and Foaling on the Chemical and Physical Composition of Milk from Kazakh Mares Kept Under Natural Pasture Conditions

by Maxat Toishimanov, Olzhas Zhanten, Rakhim Kanat, Indira Beishova, Vadim Ulyanov, Tolegen Assanbayev, Tlekbol Sharapatov, Dias Daurov, Ainash Daurova, Zagipa Sapakhova, Askar Nametov and Malika Shamekova

Animals 2025, 15(12), 1817; https://doi.org/10.3390/ani15121817 - 19 Jun 2025

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Abstract

This study aimed to evaluate the chemical composition of milk from Kazakh mares kept under free-grazing conditions on natural pasture without supplementary feeding. Fifty Kazakh mares were aged 3 to 13 years, kept in herds, and categorized by age, number of foalings, and [...] Read more.

This study aimed to evaluate the chemical composition of milk from Kazakh mares kept under free-grazing conditions on natural pasture without supplementary feeding. Fifty Kazakh mares were aged 3 to 13 years, kept in herds, and categorized by age, number of foalings, and lactation stage for statistical analysis. Milk samples were collected, chilled, and analyzed on the same day using a Milkoscan FT2. The composition of fat, protein, lactose, total solids, casein, and other components was examined. Statistical analyses, including ANOVA, PCA, CDA, and Pearson correlation analysis, were performed using JMP 17 Pro to assess the effects of lactation stage, mare age, and foaling number and to identify key relationships. The results showed that milk composition was significantly influenced by these factors. Milk from the late-lactation stage contained higher protein, lactose, and total nutrients, while milk from the early-lactation stage had a higher sugar content and acidity. Older mares and those with higher foalings produced milk with higher concentrations of glucose and galactose, while younger mares and those with fewer foalings produced milk with higher fat. A key aspect of this experiment was that all mares grazed exclusively on natural pastures without supplementation, allowing for an objective assessment of their inherent productive potential. These findings deepen the understanding of mare milk composition dynamics in Kazakh breed horses under extensive management, supporting its potential for organic dairy production. Full article

(This article belongs to the Section Equids)

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18 pages, 2339 KiB

Open AccessArticle

Effect of Adding Alkaline Metal Ions Complexes Rumen Microbiota and Metabolome of Hu Lambs

by Mingyue Li, Chi Ma, Yalin Li, Ziyi An, Yilin Yang, Feng Gao, Changqing Li and Yingchun Liu

Animals 2025, 15(12), 1816; https://doi.org/10.3390/ani15121816 - 19 Jun 2025

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Abstract

This study aimed to evaluate the effects of studying the effects of the alkali metal ion complexes (AMIC) on the rumen of lambs. Eighteen 3-month-old male Hu lambs (30 ± 2.5 kg) were randomly assigned to three groups (n = 6). Dietary [...] Read more.

This study aimed to evaluate the effects of studying the effects of the alkali metal ion complexes (AMIC) on the rumen of lambs. Eighteen 3-month-old male Hu lambs (30 ± 2.5 kg) were randomly assigned to three groups (n = 6). Dietary treatments were: control group (CG, base diet), group C1 (base diet + 0.15% AMIC), and group C2 (base diet + 0.30% AMIC). After 60 days of feeding, samples were collected for analysis. Compared with CG, rumen weight significantly increased in both C1 and C2 (p < 0.05). In C2, average daily gain (ADG), bacterial crude protein (BCP), propionic acid concentration, and rumen papillary length were significantly higher than in CG (p < 0.05). Rumen microbiota analysis showed that AMIC supplementation changed the microbial community composition, increasing the relative abundance of fiber-degrading bacteria (e.g., Prevotellaceae_UCG-001) and decreasing pathogenic Proteobacteria. In particular, rumen papillary length positively correlated with Unclassified Oscillospiraceae, Candidatus Saccharimonas, and Unclassified Clostridia vadinBB60 group. Metabolomic analysis revealed that quercetin 3-O-glucuronide levels increased in a dose-dependent manner with higher AMIC. This metabolite positively correlated with Prevotellaceae_UCG-001 abundance and ADG. At 0.30% AMIC, phospholipids PC(18:0/18:4(6Z,9Z,12Z,15Z)) and PE(18:0/16:1(9Z)) were significantly upregulated, and both positively correlated with Candidatus Saccharimonas, Unclassified Clostridia vadinBB60 group, and papillary morphology. In summary, AMIC supplementation affected metabolism by modulating the rumen microbiota, thereby promoting energy absorption and growth. The 0.30% AMIC inclusion significantly enhanced rumen papilla growth, increased the absorption area, promoted propionic acid production, reduced the acetic acid to propionic acid ratio, and ultimately improved the growth rate of Hu lambs. Thus, adding 0.30% AMIC was associated with improved growth performance. Full article

(This article belongs to the Section Small Ruminants)

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18 pages, 5239 KiB

Open AccessArticle

Monochromatic Light Impacts the Growth Performance, Intestinal Morphology, Barrier Function, Antioxidant Status, and Microflora of Yangzhou Geese

by Gang Luo, Yiyi Cheng, Yingqing Xu, Jie Liu, Wen Yang, Jiying Liu, Binbin Guo and Huanxi Zhu

Animals 2025, 15(12), 1815; https://doi.org/10.3390/ani15121815 - 19 Jun 2025

Viewed by 126

Abstract

This study investigates the effect of monochromatic light on the body weight (BW), melatonin concentration and its receptors expression levels, intestinal health, and gut microorganisms of Yangzhou geese. Green light (GL) significantly increased BW, melatonin and its receptor expression levels, villus height (VH) [...] Read more.

This study investigates the effect of monochromatic light on the body weight (BW), melatonin concentration and its receptors expression levels, intestinal health, and gut microorganisms of Yangzhou geese. Green light (GL) significantly increased BW, melatonin and its receptor expression levels, villus height (VH) and villus height/crypt depth (VH/CD) ratio, superoxide dismutase (SOD), catalase (CAT), and total antioxidant capacity (T-AOC) activities, as well as the abundance of Synergistota and Prevotellaceae_UCG-001, compared with white light (WL). Blue light (BL) significantly increased the mRNA expression of melatonin membrane receptor 1a (Mel1a) and nuclear receptor 1α (RORα), VH and VH/CD ratio, CAT activity, cecal microbes diversity, and decreased malondialdehyde (MDA) levels. Red light (RL) significantly decreased average daily feed intake, reduced the abundances of Synergistota and Prevotellaceae_UCG-001, and increased Mel1a and RORα mRNA expression levels, MDA content, and cecum microbial diversity. Moreover, melatonin levels were significantly higher in the GL and BL groups compared to RL. Furthermore, the mRNA expression levels of Claudin-10, Occludin, and occludens-1 (ZO-1) were significantly upregulated under GL or BL exposures compared to the WL group, whereas RL only enhanced the expression levels of ZO-1. Spearman’s correlation analysis revealed that the relative abundance of Prevotellaceae_UCG-001 exhibited positive correlations with BW, melatonin and its receptors expression, gut health, and antioxidant capacity. Overall, these findings suggested that GL exposure enhanced melatonin synthesis and its receptors expression, modulated intestinal homeostasis and microbial ecology, and ultimately increased goose BW. Full article

(This article belongs to the Section Poultry)

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13 pages, 400 KiB

Open AccessArticle

The In Vitro Effects of Choline on Non-Esterified Fatty Acid-Treated Bovine Peripheral Blood Leukocytes

by Cheng-Yan Li, Yueh-Tung Chen, Tossapol Moonmanee, Jacky Peng-Wen Chan and Chien-Kai Wang

Animals 2025, 15(12), 1814; https://doi.org/10.3390/ani15121814 - 19 Jun 2025

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Abstract

The transition period is defined as 3 weeks around parturition, involving the rapid increase in the energy demand, promoting adipose mobilization and non-esterified fatty acid (NEFA) release. High NEFA levels might cause oxidative stress and associated health risks, including the disruption of the [...] Read more.

The transition period is defined as 3 weeks around parturition, involving the rapid increase in the energy demand, promoting adipose mobilization and non-esterified fatty acid (NEFA) release. High NEFA levels might cause oxidative stress and associated health risks, including the disruption of the immune capability of peripheral leukocytes. Nutrient supplementation of choline, which improves cellular lipid metabolism and controls lipid oxidation, potentially maintains the integrity of peripheral leukocytes and alleviates the impacts of increased NEFAs. This study investigated the effects of choline on bovine peripheral blood leukocytes (PBLs) treated with high levels of NEFAs. Peripheral blood mononuclear cells (PBMCs) and polymorphonuclear leukocytes (PMNs) were isolated from dry cows, and treated with 1 mM NEFA in combination with 0, 4, or 12 μM choline. The expression of pro-inflammatory cytokines and oxidative stress indicators was determined. This study demonstrates that 1 mM NEFA induces lipid oxidation and pro-inflammatory cytokine expression in PBLs. Supplementation with 4 μM and 12 μM choline significantly reduced NEFA-induced lipid oxidation; however, it did not affect pro-inflammatory cytokine mRNA expression. In conclusion, choline supplementation may help alleviate NEFA-induced oxidative stress in bovine peripheral blood leukocytes, highlighting its potential as a functional supplement during the transition period. However, it had no observable effect on mitigating pro-inflammatory responses, indicating that additional strategies may be needed to address NEFA-induced immune activation. Full article

(This article belongs to the Section Cattle)

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15 pages, 1261 KiB

Open AccessArticle

Functional Requirement of Niacinamide for Blood Profiles, Antioxidant Status, and Intestinal Health in Finishing Pigs Fed a Low-Protein Diet

by Yan Zhao, Fangli Tang, Yunlong Shi, Qinyu Tan, Qingxin Ju, Ziyi Yang, Guanqing Yang, Pengfei Gao, Sung Woo Kim, Lin Xi, Guoqing Cao and Bugao Li

Animals 2025, 15(12), 1813; https://doi.org/10.3390/ani15121813 - 19 Jun 2025

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Abstract

This study investigated the effects of dietary niacinamide supplementation on blood parameters, antioxidant status, and intestinal health in finishing pigs fed low-protein diets. Sixty-four pigs (Duroc × Landrace × Yorkshire; 80.4 ± 0.1 kg) were randomly allocated to four dietary treatments supplemented with [...] Read more.

This study investigated the effects of dietary niacinamide supplementation on blood parameters, antioxidant status, and intestinal health in finishing pigs fed low-protein diets. Sixty-four pigs (Duroc × Landrace × Yorkshire; 80.4 ± 0.1 kg) were randomly allocated to four dietary treatments supplemented with 30 (NAM30), 130 (NAM130), 230 (NAM230), and 330 (NAM330) mg/kg niacinamide for 30 days. Each treatment had four replicate pens and four pigs per pen. Growth performance was not significantly affected. However, the NAM130 group showed higher (p < 0.05) hemoglobin levels, reduced (p < 0.05) serum malondialdehyde and interleukin-1β (IL-1β) concentrations, and altered intestinal microbiota composition, including lower Streptococcus abundance (p < 0.05). Serum alanine aminotransferase levels increased quadratically (p < 0.05) with niacinamide supplementation, with a breakpoint at approximately 221 mg/kg. These results suggest that dietary supplementation with 130 mg/kg niacinamide improves antioxidant status, modulates inflammation, and supports intestinal microbial balance, with a safety threshold to avoid hepatic stress. Full article

(This article belongs to the Section Animal Nutrition)

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23 pages, 4983 KiB

Open AccessArticle

Post Mortem Findings of Cetaceans Stranded Along the Campania Coast from 2016 to 2022

by Emanuele Esposito, Maria Oliviero, Doriana Iaccarino, Gianluigi Paduano, Francesco Serra, Martina Levante, Maria Grazia Amoroso, Clementina Auriemma, Amalia Gallo, Maria Gabriella Lucibelli, Agata Campione, Roberta Rispoli, Francesca Menafro, Francesca Bove, Maria Dimatteo, Marianna D’Amore, Barbara Degli Uberti, Virginia Mattioda, Federica Giorda, Carla Grattarola, Guido Pietroluongo, Cinzia Centelleghe, Giovanna Fusco, Esterina De Carlo and Fabio Di Nocera Show full author list Hide full author list

Animals 2025, 15(12), 1812; https://doi.org/10.3390/ani15121812 - 19 Jun 2025

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Abstract

The presence of cetaceans along the Campania coast has always been documented. Between 2016 and 2022, out of 65 cetaceans stranded along the Campania coast, 46 were studied for bacteriological, virological, parasitological, and histopathological investigations. The results highlighted that for 59% (n = [...] Read more.

The presence of cetaceans along the Campania coast has always been documented. Between 2016 and 2022, out of 65 cetaceans stranded along the Campania coast, 46 were studied for bacteriological, virological, parasitological, and histopathological investigations. The results highlighted that for 59% (n = 27) of the specimens, the cause of death was of natural origin, while for only 2% (n = 1) of animals, the origin of death was ascribed to anthropic causes. Unfortunately, for 39% (n = 18) of the cetaceans, it was impossible to determine the cause of death. All the cetaceans that died of natural causes showed viral, bacterial, and parasitic infections. The primary pathogens detected were Cetacean Morbillivirus (CeMV, 65.2%, n = 30/46), Toxoplasma gondii (10.9%, n = 5/46), and Brucella ceti (8.7%, n = 4/46). The animals showed typical lesions of the isolated pathogens, such as systemic infection, meningoencephalitis, and pneumonia. Moreover, even with a lower frequency, other relevant pathogens like Photobacterium damselae, Salmonella enteritidis, Erysipelothrix rhusiopathiae, and Chlamydia abortus were isolated. These data were useful to understand the spread and circulation of these pathogens, some zoonotic, in the coastal marine waters of the Campania region. Full article

(This article belongs to the Special Issue Wildlife Histopathology and Molecular Diagnosis)

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15 pages, 6589 KiB

Open AccessArticle

Preclinical Evaluation of Fenbendazole for Controlling Gyrodactylus kobayashii (Monogenea, Gyrodactylidae) in Goldfish: Dose Optimization and Safety Assessment

by Jing Dong, Jiangtao Li, Yongtao Liu, Qiuhong Yang, Ning Xu, Xiaohui Ai and Shun Zhou

Animals 2025, 15(12), 1811; https://doi.org/10.3390/ani15121811 - 19 Jun 2025

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Abstract

This preclinical study investigated the efficacy and safety of fenbendazole, a broad-spectrum benzimidazole anthelmintic, for the treatment of Gyrodactylus kobayashii in goldfish (Carassius auratus). In vivo bath treatments demonstrated potent, dose-dependent anthelmintic efficacy, achieving 98.58% efficacy at a concentration of 0.02 [...] Read more.

This preclinical study investigated the efficacy and safety of fenbendazole, a broad-spectrum benzimidazole anthelmintic, for the treatment of Gyrodactylus kobayashii in goldfish (Carassius auratus). In vivo bath treatments demonstrated potent, dose-dependent anthelmintic efficacy, achieving 98.58% efficacy at a concentration of 0.02 mg/L and a 48 h EC₅₀ of 0.006 mg/L. A short-duration (6 h) bath at 0.06 mg/L, followed by an 18 h recovery period in dechlorinated water, resulted in complete parasite elimination. However, acute toxicity assay indicated a relatively narrow safety margin for prolonged bath treatments, with a 96 h LC₅₀ of 0.039 mg/L, highlighting the need for caution when employing extended bath treatments. Oral administration of fenbendazole at 20 mg/kg body weight for three consecutive days resulted in an efficacy of 83.35%, which increased to 96.28% by seven days post-treatment. Safety evaluations revealed this regimen induced transient oxidative stress and mild, reversible histopathological alterations in the liver and gills. Biochemical and histological markers indicated a recovery trend, approaching baseline levels by 15 days post-treatment. These findings suggested that oral fenbendazole is an effective and relatively safe anthelmintic treatment against G. kobayashii in goldfish. This study underscores the potential of drug repurposing as an effective strategy for developing novel anthelmintic agents in aquaculture. Full article

(This article belongs to the Special Issue Aquatic Animal Medicine and Pathology)

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12 pages, 1364 KiB

Open AccessArticle

Preliminary Study of the Antimicrobial Capacity of the Cutaneous Mucus and Smear Cytology of the Epidermis in a Population of European eels (Anguilla anguilla, Linnaeus 1758)

by Enrico Volpe, Sara Ciulli, Maria Morini, Laura Gentile, Antonio Casalini, Chiara Gentilezza and Luciana Mandrioli

Animals 2025, 15(12), 1810; https://doi.org/10.3390/ani15121810 - 19 Jun 2025

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Abstract

The skin and its products, such as the mucus, represent an important defense mechanism against infection by pathogens. Various environmental conditions can alter mucus composition and/or cells embedded in this matrix. The analysis of skin mucus is considered a useful method to evaluate [...] Read more.

The skin and its products, such as the mucus, represent an important defense mechanism against infection by pathogens. Various environmental conditions can alter mucus composition and/or cells embedded in this matrix. The analysis of skin mucus is considered a useful method to evaluate the biological response of fish to stimuli. The mucus and cells can be considered suitable non-invasive biomarkers. With this preliminary study, an antibacterial capacity method was applied to cutaneous mucus samples of a European eel population. This standardized method highlighted the ability of the mucus to significantly counteract the proliferation of two bacteria, the opportunistic pathogen Aeromonas hydrophila and the primary pathogen Aeromonas salmonicida. The cytological method using skin scraping has proven to be a useful non-invasive tool, having allowed the highlighting of the two most represented cellular populations of the cutaneous covering, the epidermal cells and the goblet cells, in combination with mucous strands. Cytological findings did not disclose pathologic elements in the mucus. A broader use of these two non-invasive microbiological and cytological methods can provide useful information on fish health, indirectly contributing to the conservation of the species. Full article

(This article belongs to the Section Aquatic Animals)

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10 pages, 735 KiB

Open AccessArticle

Assessment of Stress in Dogs Under Cancer Therapy via Faecal Cortisol Metabolite Analysis: A Pilot Study

by Christina Ziegerhofer, Alexander Tichy, Miriam Kleiter, Birgitt Wolfesberger and Rupert Palme

Animals 2025, 15(12), 1809; https://doi.org/10.3390/ani15121809 - 19 Jun 2025

Viewed by 183

Abstract

Stress reactions play an important role in animals’ ability to cope with various situations. Glucocorticoids are measured as a stress parameter, and analysis of their faecal metabolites has proven to be a good method for evaluating long term stress. We hypothesised that dogs [...] Read more.

Stress reactions play an important role in animals’ ability to cope with various situations. Glucocorticoids are measured as a stress parameter, and analysis of their faecal metabolites has proven to be a good method for evaluating long term stress. We hypothesised that dogs suffering from cancer would have a higher stress level during cancer therapy, which would be reflected in higher levels of faecal cortisol metabolites (FCMs). Faeces were collected from 40 dogs receiving radiation or chemotherapy before and during the treatment, and from 53 healthy dogs, who served as a control group. FCMs were extracted and quantified by a cortisol enzyme immunoassay. The results showed that the stress levels were not significantly higher in the cancer patients before therapy compared to those in the control group. Additionally, there were no significant differences in the FCM concentrations of dogs at specific time points during chemotherapy or radiotherapy. Therefore, suspected stress should not be a criterion to exclude animals from cancer treatment. Such a treatment seems ethically justifiable if it is expected to provide benefits and improvement in the quality of life for patients. Full article

(This article belongs to the Section Veterinary Clinical Studies)

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18 pages, 931 KiB

Open AccessReview

Sperm Membrane: Molecular Implications and Strategies for Cryopreservation in Productive Species

by Macarena Castro, Karla Leal, Felipe Pezo and María José Contreras

Animals 2025, 15(12), 1808; https://doi.org/10.3390/ani15121808 - 19 Jun 2025

Viewed by 258

Abstract

Sperm cryopreservation is a fundamental reproductive biotechnology, enabling the long-term storage of genetic material and supporting assisted reproduction programs. Despite its widespread application, the process is associated with significant limitations due to the cryo-induced cellular damage that occurs during freezing and thawing. These [...] Read more.

Sperm cryopreservation is a fundamental reproductive biotechnology, enabling the long-term storage of genetic material and supporting assisted reproduction programs. Despite its widespread application, the process is associated with significant limitations due to the cryo-induced cellular damage that occurs during freezing and thawing. These injuries primarily affect the plasma membrane, nuclear DNA, and motility, thereby compromising the fertilizing potential of spermatozoa. Furthermore, interspecies variability in terms of cryo-sensitivity poses a major challenge to the development of standardized cryopreservation protocols. Recent advances have focused on mitigating cryodamage through the use of various strategies. The inclusion of antioxidants in cryopreservation media has proven effective in reducing oxidative stress, thereby enhancing cellular protection. Similarly, the addition of lipid-based supplements contributes to membrane stabilization, improving post-thaw sperm viability and functionality. Moreover, the application of omics technologies, such as transcriptomics and proteomics, has facilitated a deeper understanding of molecular damage and protective responses, paving the way for the development of tailored, species-specific protocols. These integrated approaches optimize cryopreservation conditions, maximizing post-thaw survival and the fertilizing capacity of sperm. Enhancing cryopreservation techniques not only improves the outcomes of assisted reproductive technologies, but also plays a crucial role in the conservation of genetically valuable livestock species. In conclusion, the integration of biotechnological and molecular tools holds significant promise for overcoming the current limitations and advancing the efficacy of sperm cryopreservation. Full article

(This article belongs to the Special Issue Current Status and Advances in Semen Preservation—Second Edition)

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13 pages, 3435 KiB

Open AccessArticle

Transcriptomic Analysis of Tambaqui (Colossoma macropomum) Exposed to Trichlorfon-Induced Toxicity

by Hallana Cristina Menezes da Silva, Igor Kelvyn Cavalcante Lobo, André Gentil da Silva, Ana Lúcia Silva Gomes, Wallice Paxiúba Duncan, Juliana Costa Silva, Fabrício M. Lopes, Roberto Ferreira Artoni and Daniele Aparecida Matoso

Animals 2025, 15(12), 1807; https://doi.org/10.3390/ani15121807 - 19 Jun 2025

Viewed by 136

Abstract

Trichlorfon is an antiparasitic agent widely used to control pests and parasites in farmed fish. Tambaqui (C. macropomum) is the most commercially important characid species farmed in the Amazon region. Trichlorfon exposure is known to cause physiological damage in fish due [...] Read more.

Trichlorfon is an antiparasitic agent widely used to control pests and parasites in farmed fish. Tambaqui (C. macropomum) is the most commercially important characid species farmed in the Amazon region. Trichlorfon exposure is known to cause physiological damage in fish due to its organophosphate nature. In this study, we used RNA-Seq to investigate the hepatic response of tambaqui following exposure to 0.435 mg/L of trichlorfon. The analysis revealed activation of several metabolic pathways, particularly those related to tumor processes, immune responses, and apoptosis. Additionally, we identified upregulation of solute carrier (SLC) genes, which may facilitate trichlorfon entry into hepatocytes. These findings enhance our understanding of fish responses to antiparasitic agents and support further research into the molecular impacts of organophosphate compounds in aquaculture species. Full article

(This article belongs to the Special Issue Ecotoxicology in Aquatic Animals: 2nd Edition)

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14 pages, 1238 KiB

Open AccessArticle

Optimization of Solid Lipid Microcapsule Matrix for Enhanced Release and Bioavailability of L-Lysine in Swine

by Costanza Bonnici, Maria Federica Marchesi, Martina Felici, Federico Ghiselli, Roberta Majer, Benedetta Tugnoli, Guglielmo Gallina, Andrea Piva and Ester Grilli

Animals 2025, 15(12), 1806; https://doi.org/10.3390/ani15121806 - 19 Jun 2025

Viewed by 170

Abstract

L-lysine (L-Lys) is the first-limiting amino acid in swine nutrition, but free-form supplements exhibit poor intestinal absorption, reducing their bioavailability. This study aimed to enhance the gastric retention, controlled intestinal release, and systemic availability of L-Lys by optimizing solid lipid microcapsules (SLMs). SLMs [...] Read more.

L-lysine (L-Lys) is the first-limiting amino acid in swine nutrition, but free-form supplements exhibit poor intestinal absorption, reducing their bioavailability. This study aimed to enhance the gastric retention, controlled intestinal release, and systemic availability of L-Lys by optimizing solid lipid microcapsules (SLMs). SLMs were formulated using hydrogenated triglycerides (C16:0 or C18:1), free fatty acids, and varying emulsifier concentrations. Gastric retention and intestinal release were evaluated in vitro under simulated gastrointestinal conditions (a pepsin buffer at pH 5.0 for 2 h, followed by a pancreatin buffer at pH 6.5 for up to 8 h at 39 °C). SLMs with hydrogenated triglycerides showed significantly higher gastric retention (94–95%) than those with free fatty acids (48%). Specifically, C16:0 triglyceride-based SLMs achieved 74% intestinal release, which was enhanced to 90% with 1% emulsifier. This refined formulation was subsequently evaluated in vivo using weaned pigs (three groups; n = 4) fed a basal cornmeal diet. The treatments included a single oral administration of saline solution (placebo), free L-Lys (0.17 g/kg BW), or L-Lys SLMs (0.38 g/kg BW, equally providing L-Lys at 0.17 g/kg BW). The SLMs delayed the L-Lys plasma peak (T. max. 3–4 h vs. 1 h) and significantly increased the total L-Lys amount in the plasma over 24 h, demonstrating the enhanced relative bioavailability of encapsulated L-Lys. Full article

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11 pages, 250 KiB

Open AccessArticle

Effect of Freezing for up to 120 Days on the Physicochemical Characteristics of Hamburgers Made from Botucatu Rabbit Does Slaughtered at Different Ages

by Erick Alonso Villegas-Cayllahua, Daniel Rodrigues Dutra, Ana Veronica Lino Dias, Thamiris Daiane Domenici, Leandro Dalcin Castilha and Hirasilva Borba

Animals 2025, 15(12), 1805; https://doi.org/10.3390/ani15121805 - 19 Jun 2025

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Abstract

This study evaluated the effect of freezing for up to 120 days on the physicochemical and technological properties of hamburgers made from Botucatu rabbit does slaughtered at 3, 12, and 24 months of age. The parameters were evaluated as follows: surface color ( [...] Read more.

This study evaluated the effect of freezing for up to 120 days on the physicochemical and technological properties of hamburgers made from Botucatu rabbit does slaughtered at 3, 12, and 24 months of age. The parameters were evaluated as follows: surface color (L*, a*, b*), pH using an insertion pH meter, cooking loss using a grill, storage loss based on weight differences, shear force in cooked samples using a texture analyzer, shrinkage percentage, chemical composition (moisture, protein, lipids, and ash), and lipid oxidation, determined by measuring the concentration of malondialdehyde in the burgers at different storage intervals (0, 60, and 120 days) under freezing conditions (−18 °C). The results indicated that increased storage time and animal age reduced tenderness and increased lipid content (p < 0.05). Burgers made from younger does showed higher levels of lipid oxidation. Age also influenced color (greater redness and lower lightness in older animals) and chemical composition, with older does producing burgers with higher protein and lower moisture and mineral content. However, all samples remained within the limits established by Brazilian legislation. This study recommends using meat from does of different ages for hamburger production, as all variations met the required legal standards. Full article

(This article belongs to the Special Issue Animal Breeds and Muscle Growth Potential: Opportunities for Sustainable Farming)

19 pages, 352 KiB

Open AccessArticle

Influence of Dietary Alpha-Tocopheryl Acetate (Vitamin E) and Animal Fat on the Chemical Composition, Fatty Acid Profile, Lipid Stability and Sensory Traits of Fresh and Stored Hamburgers from Rabbit Meat

by Bianca Palumbo, Maria Elena Cossu and Antonella Dalle Zotte

Animals 2025, 15(12), 1804; https://doi.org/10.3390/ani15121804 - 19 Jun 2025

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Abstract

Improving the proportion of polyunsaturated fatty acids (PUFAs) in meat enhances its nutritional value; however, because PUFAs are prone to oxidation, incorporating antioxidants into animal diets may be an effective way to limit lipid degradation. This study investigated the effects of dietary fat [...] Read more.

Improving the proportion of polyunsaturated fatty acids (PUFAs) in meat enhances its nutritional value; however, because PUFAs are prone to oxidation, incorporating antioxidants into animal diets may be an effective way to limit lipid degradation. This study investigated the effects of dietary fat content, vitamin E supplementation, and storage time on the oxidative stability, physico-chemical, sensory, and technological traits of rabbit hamburgers. Seventy-two rabbits (aged 40–78 days) were assigned to four groups: F0-E0 (0% pork fat, 0 mg/kg vitamin E), F0-E200 (0% fat, 200 mg/kg vitamin E), F2-E0 (2% fat, 0 mg/kg vitamin E), and F2-E200 (2% fat, 200 mg/kg vitamin E). After slaughter and carcass deboning, fat-free meat was processed into hamburgers. Three hamburgers per rabbit were analyzed immediately (T0) and three after 7 days of storage at 4 °C (T7). Vitamin E supplementation significantly reduced TBARS values (p < 0.001) and improved meat appearance and texture. Fat-enriched diets modified the fatty acid profile, increasing unsaturated and n-3 fatty acids, and enhanced meat flavor. Vitamin E supplementation improved the nutritional quality of the lipids by increasing PUFAs and n-3 FA levels. Storage time significantly affected quality traits, with T7 hamburgers showing higher TBARS values (p < 0.001) and a greater UFA percentage. Full article

(This article belongs to the Section Animal Nutrition)

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3 pages, 161 KiB

Open AccessCorrection

Correction: Mertz et al. Interdisciplinary Animal Research Ethics—Challenges, Opportunities, and Perspectives. Animals 2024, 14, 2896

by Marcel Mertz, Tatiana Hetzel, Karla Alex, Katharina Braun, Samuel Camenzind, Rita Dodaro, Svea Jörgensen, Erich Linder, Sara Capas-Peneda, Eva Ingeborg Reihs, Vini Tiwari, Zorana Todorović, Hannes Kahrass and Felicitas Selter

Animals 2025, 15(12), 1803; https://doi.org/10.3390/ani15121803 - 19 Jun 2025

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Abstract

In the original version [...] Full article

14 pages, 940 KiB

Open AccessArticle

The Influence of Genotype and Seasonality on the Sow Colostrum Quality and Immunoglobulin G Content

by Kristina Gvozdanović, Vice Čuljak, Vladimir Margeta, Ivona Djurkin Kušec, Boris Antunović, Dalida Galović and Goran Kušec

Animals 2025, 15(12), 1802; https://doi.org/10.3390/ani15121802 - 18 Jun 2025

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Abstract

The aim of this study was to investigate the influence of genotype (GT) and seasonality (NP) on the quality parameters of sow colostrum and evaluate the efficiency of the radial immunodiffusion (RID) analysis and the Brix refractometer in determining the IgG concentration. This [...] Read more.

The aim of this study was to investigate the influence of genotype (GT) and seasonality (NP) on the quality parameters of sow colostrum and evaluate the efficiency of the radial immunodiffusion (RID) analysis and the Brix refractometer in determining the IgG concentration. This study was conducted on 240 sows that originated from two genotypes, namely GT1 (TOPIGS, n = 120) and GT2 (Pig Improvement Company, n = 120), during the three farrowing periods: the winter farrowing period (WNP, n = 80), the summer farrowing period (SMP, n = 80) and the spring farrowing period (SSP, n = 80). The significant interaction effect was observed for protein (p < 0.0001), lactose (p < 0.05) and non-fat solids (SNT) (p < 0.001). At the same time, the interaction effect influenced the IgG concentration measured with the Brix refractometer (p < 0.0001) and RID (p < 0.0001). Pearson’s correlation coefficient showed that Brix percentage was positively correlated with RID results (r = 0.52, p < 0.0001), while the Bland–Altman plots indicated a mean bias of −1.93. Partial eta-squared analysis (η²) showed that the genotype explained the largest proportion of variance in fat content (η² = 0.136) and IgG concentration (η² = 0.164), while interaction effects were largest for protein (η² = 0.072). The results of this study show that genotype and seasonality influence sow colostrum quality, which indicates the importance of genotype−seasonality interactions in breeding programs for optimizing the colostrum quality and piglet survival. Full article

(This article belongs to the Special Issue Genetic Research for Improving Livestock Heat Stress Resistance)

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11 pages, 874 KiB

Open AccessArticle

Feline Leishmaniosis: A Retrospective Study of Seroprevalence in Cats in the Campania Region, Southern Italy

by Laura Cortese, Giulia Abate, Pasquale Santoro, Elvira Improda, Gianmarco Ferrara, Vincenzo Lucidi, Antonio Sica, Giuseppe Iovane and Serena Montagnaro

Animals 2025, 15(12), 1801; https://doi.org/10.3390/ani15121801 - 18 Jun 2025

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Abstract

Feline leishmaniasis (FeL), caused by Leishmania infantum, is increasingly reported in areas of endemic Mediterranean canine leishmaniasis (CanL), making it an emerging feline disease. This cross-sectional study investigated L. infantum seroprevalence and risk factors in 229 domestic cats from the Campania region [...] Read more.

Feline leishmaniasis (FeL), caused by Leishmania infantum, is increasingly reported in areas of endemic Mediterranean canine leishmaniasis (CanL), making it an emerging feline disease. This cross-sectional study investigated L. infantum seroprevalence and risk factors in 229 domestic cats from the Campania region of southern Italy, a CanL endemic area, between January 2023 and December 2024. Serum samples were tested for L. infantum antibodies (IFAT) and for FIV/FeLV. Seropositivity (IFAT titre ≥ 1:40) for FeL was detected in 12/229 (5.2%) of the cats tested. No statistically significant correlation was found between seropositivity for L. infantum and the variables considered. However, outdoor cats and FIV/FeLV-seropositive cats had higher prevalence rates: 10.6% and 7.4%, respectively. Of the 12 seropositive cats, 7 (58.3%) had an antibody titre of 1:40, 2 (16.6%) of 1:80 and 3 (25.0%) a titre of 1:160. Of the 12 cats positive for FeL, 2 (16.6%) were also positive for FIV. Our results confirm the exposure to L. infantum and the serological response in cats from southern Italy. The low prevalence could be due to owners using mosquito control products in the household that would also protect cats. Further investigation is essential to clarify risk factors and improve our understanding of the epidemiology of FeL in this endemic area. Full article

(This article belongs to the Section Companion Animals)

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18 pages, 2173 KiB

Open AccessArticle

Effects of Dietary Protein Levels on the Growth, Physiological, and Biochemical Indices of Juvenile Yellow River Carp (Cyprinus carpio haematopterus)

by Xiaona Jiang, Feihu Qu, Yanlong Ge, Chitao Li, Xiaodan Shi, Xuesong Hu, Lei Cheng, Xinyu Zhao and Zhiying Jia

Animals 2025, 15(12), 1800; https://doi.org/10.3390/ani15121800 - 18 Jun 2025

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Abstract

An appropriate protein content in the diet can effectively increase the growth rate, muscle quality, and environmental stress resistance of fish. In this study, juvenile Yellow River carp (51.56 ± 0.17 g) were fed isofat diets with different protein concentrations (22%, 25%, 28%, [...] Read more.

An appropriate protein content in the diet can effectively increase the growth rate, muscle quality, and environmental stress resistance of fish. In this study, juvenile Yellow River carp (51.56 ± 0.17 g) were fed isofat diets with different protein concentrations (22%, 25%, 28%, 31%, 34%, and 37%). The results showed that, compared with other protein content groups, when the protein content was 34%, the WGR, SGR, and FCR were significantly higher, while the FCR was significantly lower (p < 0.05). Among them, the SGR and FCR were positively correlated with the dietary protein content, with the regression equations being y₁ = −32.208x² + 21.897x − 1.4001 (R² = 0.8622) and y₂ = 97.027x² − 68.428x + 13.269 (R² = 0.9663), respectively. When x was 33.99% and 35.26%, the SGR and FCR had extreme values. The contents of CP, Lys, EAA, and EAA/TAA were significantly greater in the 34% protein group, reflecting muscle quality (p < 0.05). In addition, the activities of α-AMS, LPS, TPS, SOD, CAT, and GSH-Px were significantly greater in the 34% protein group (p < 0.05). Similarly, the relative expression levels of GH, IGF-I, TOR, 4EBP2, Rhag, Rhbg, and Rhcg1 were significantly greater in the 34% protein group compared to the other protein groups (p < 0.05). The above results indicated that when the protein content in the diet was 34%, it significantly improved the growth and stress resistance of juvenile Yellow River carp. Based on the nonlinear regression equations for the SGR and FCR, the optimal dietary protein content of juvenile Yellow River carp (51.56 ± 0.17 g) was determined to be 33.99–35.26%. Full article

(This article belongs to the Special Issue Dietary Components in Animal Nutrition: Favoring Sustainability, Welfare and Safety)

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22 pages, 2163 KiB

Open AccessArticle

Status of Selenium and Other Essential and Toxic Elements in Oregon Grazing Sheep

by Daniella Hasan, Christopher J. Russo, Katherine R. McLaughlin, Gene Pirelli and Massimo Bionaz

Animals 2025, 15(12), 1799; https://doi.org/10.3390/ani15121799 - 18 Jun 2025

Viewed by 318

Abstract

Mineral imbalances in sheep can have significant health and economic consequences, yet regional assessments of trace mineral and toxic element status are limited. Oregon is an endemically selenium (Se)-deficient region, but a statewide evaluation has not been conducted. This study assessed whole blood [...] Read more.

Mineral imbalances in sheep can have significant health and economic consequences, yet regional assessments of trace mineral and toxic element status are limited. Oregon is an endemically selenium (Se)-deficient region, but a statewide evaluation has not been conducted. This study assessed whole blood concentrations of 18 elements in 370 clinically normal ewes from 56 farms across Oregon to determine the prevalence of deficiencies and potential toxic exposures. A deficiency threshold of <120 ng Se/mL whole blood was used. We found that 28% of animals were Se-deficient, and 27% of farms had an average whole blood Se below this threshold. No animals reportedly showed overt clinical signs of deficiency or toxicity, and no excessive exposure to toxic elements such as arsenic, lead, or cadmium was found. We also compared mineral concentrations in blood collected using standard EDTA tubes versus mineral-free EDTA tubes, and in whole blood versus plasma, to assess alternative sampling strategies. Strong correlations (r > 0.9) for Se, cobalt (Co), and molybdenum (Mo) between plasma and whole blood allowed for the derivation of conversion equations. These findings provide updated insights into the mineral status of Oregon sheep, contribute to whole blood reference data, and support practical alternatives to improve trace mineral testing in field conditions. Full article

(This article belongs to the Section Small Ruminants)

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11 pages, 667 KiB

Open AccessArticle

Optimization of Black Boar Sperm Cryopreservation Efficiency with Antioxidant-Rich Plant Extracts from Djulis (Chenopodium formosanum)

by Wenchi Hsu, Tzuche Lin, Shenchang Chang, Minjung Lin, Chaowei Huang, Perngchih Shen, Chihjen Chou and Shaoyu Peng

Animals 2025, 15(12), 1798; https://doi.org/10.3390/ani15121798 - 18 Jun 2025

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Abstract

The cryopreservation of boar sperm effectively extends its storage period but often leads to increased reactive oxygen species (ROS) production, compromising sperm quality. Plant extracts, rich in bioactive compounds such as polyphenols and flavonoids, have been shown to reduce ROS. Djulis (Chenopodium [...] Read more.

The cryopreservation of boar sperm effectively extends its storage period but often leads to increased reactive oxygen species (ROS) production, compromising sperm quality. Plant extracts, rich in bioactive compounds such as polyphenols and flavonoids, have been shown to reduce ROS. Djulis (Chenopodium formosanum), also known as the “ruby of cereals”, is nutritionally rich and holds potential as a cryoprotective additive. This study aimed to determine the optimal concentration of extracts from different parts of djulis, including unhulled seeds and stems, for effective boar semen cryopreservation. Fresh semen from Taiwan indigenous boars was diluted with a modified GLT-cryoprotectant extender containing glycerol, low-density lipoprotein (LDL), and trehalose. The experimental groups included DSS25, DSS50, DS25, and DS50—representing djulis unshelled seed at 25 mg/mL and 50 mg/mL, and djulis stem at 25 mg/mL and 50 mg/mL in distilled water, respectively—alongside a control group without additives. Post-thaw assessments included sperm motility, kinetic parameters, viability, acrosome integrity, and the antioxidant properties of djulis extracts, such as DPPH radical scavenging activity and total phenolic acid content. Results showed that total motility (TM) was significantly higher in the DSS25 (48.8 ± 3.9), DSS50 (49.0 ± 6.7), and DS50 (49.0 ± 2.4) groups compared to the control group (31.3 ± 4.8). Similarly, progressive motility (PM) was significantly improved in DSS25 (27.5 ± 2.7) and DSS50 (26.8 ± 4.1) versus the control (12.8 ± 3.2). However, for straightness (STR), the control group (87.8 ± 1.3) exhibited significantly higher values than the DS50 group (83.5 ± 1.3) (p < 0.05). Viability and acrosome integrity showed no significant differences across groups. In conclusion, djulis extracts positively influence sperm motility and forward movement, with 1% djulis extract confirmed to enhance the quality of cryopreserved semen. Future research will focus on determining the optimal dosage of djulis extract for improved cryopreservation outcomes. Full article

(This article belongs to the Section Animal Reproduction)

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12 pages, 570 KiB

Open AccessArticle

Objective Evaluation of Gait Asymmetries in Traditional Racehorses During Pre-Race Inspection: Application of a Markerless AI System in Straight-Line and Lungeing Conditions

by Federica Meistro, Maria Virginia Ralletti, Riccardo Rinnovati and Alessandro Spadari

Animals 2025, 15(12), 1797; https://doi.org/10.3390/ani15121797 - 18 Jun 2025

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Abstract

Subtle locomotor asymmetries are common in horses and may go unnoticed during routine pre-race clinical inspections, particularly when based solely on subjective evaluation. This study aimed to describe vertical head and pelvic movement asymmetries in racehorses that passed official pre-race inspections at a [...] Read more.

Subtle locomotor asymmetries are common in horses and may go unnoticed during routine pre-race clinical inspections, particularly when based solely on subjective evaluation. This study aimed to describe vertical head and pelvic movement asymmetries in racehorses that passed official pre-race inspections at a traditional racing event. Twenty-four horses were analysed using a markerless AI-based gait analysis system while trotting in-hand and during lungeing in both directions. Asymmetry parameters (HDmin, HDmax, PDmin, and PDmax) were extracted from video recordings, with values ≥0.5 considered clinically relevant. Vertical asymmetries were detected in 71% of horses during straight-line evaluation and in 79% during at least one lungeing direction. Some horses showed relevant asymmetries only under specific movement conditions, underscoring the complementary role of straight-line and lungeing assessments in comprehensive gait evaluation. These results suggest that objective gait analysis could enhance pre-race veterinary assessments, especially in traditional racing, where horses are subjected to significant biomechanical stress, including variable surface properties and repetitive directional loading. In such complex and dynamic environments, relying solely on visual assessment may result in the underdiagnosis of subtle locomotor alterations. The AI-based tools offer potential to improve the detection of subtle irregularities and support evidence-based decisions in performance horse management. Further investigations are warranted to validate the clinical relevance of currently adopted asymmetry thresholds, refine their diagnostic value, and support their integration into standardized pre-race evaluation protocols. Full article

(This article belongs to the Section Equids)

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13 pages, 5692 KiB

Open AccessArticle

The Thoroughbred Theory: Influence of Breed on Performance at the CCI5*-L Level of Eventing

by Kianna R. Walz, Meghan E. McCormick and Carleigh E. Fedorka

Animals 2025, 15(12), 1796; https://doi.org/10.3390/ani15121796 - 18 Jun 2025

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Abstract

Breed bias exists within many of the top disciplines of equestrian sport. Therefore, the objective of this study was to evaluate the influence of breed on performance at the CCI5*-L level of eventing. To do so, the results of all CCI5*-L events from [...] Read more.

Breed bias exists within many of the top disciplines of equestrian sport. Therefore, the objective of this study was to evaluate the influence of breed on performance at the CCI5*-L level of eventing. To do so, the results of all CCI5*-L events from 2014 to 2024 were assessed. Initially, Thoroughbreds were compared to all non-Thoroughbreds, and this was followed by assessing the ten primary breeds. Data included penalties accrued in dressage, cross country, show jumping, and overall. Additionally, the likelihood of completing each phase was assessed. It was found that Thoroughbreds accrued more dressage penalties than non-Thoroughbreds (p < 0.001). When comparing primary breeds, Thoroughbreds were found to accumulate more dressage penalties than Hannoverian, Irish Sport Horse, Dutch Warmblood, Oldenburg, and Selle Francais (p < 0.03). When assessing cross-country, Thoroughbreds were more likely to finish without fault when compared to non-Thoroughbreds (p < 0.001). Non-Thoroughbreds were more likely to show jump without fault (p < 0.001), while Thoroughbred were found to have higher overall show jumping penalties in comparison to Anglo-European Studbook, Holsteiner, Irish Sport Horse, Dutch Warmblood, Oldenburg, and Selle Francais (p = 0.01). When evaluating overall penalties accumulated throughout the event, there was no effect of breed (p = 0.42). Overall, there was no advantage in competing a non-Thoroughbred in comparison to a Thoroughbred when considering total penalties accrued, indicating a level playing field within competition. Therefore, breed bias against TBs should be negated when selecting for CCI5*-L eventing prospects. Full article

(This article belongs to the Section Equids)

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11 pages, 2131 KiB

Open AccessCase Report

Case of Japanese Marten (Martes melampus) Identification by mtDNA Analysis in a Series of Vehicle Cable Damage Incidents

by Reina Ueda, Yuko Kihara, Shin-ichi Hayama and Aki Tanaka

Animals 2025, 15(12), 1795; https://doi.org/10.3390/ani15121795 - 18 Jun 2025

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Abstract

A series of incidents involving damage to vehicle speed sensor cables occurred in an urban area in Japan. At the request of the police, DNA analysis was conducted to identify the animal species responsible. Swab samples collected from the damaged sections of the [...] Read more.

A series of incidents involving damage to vehicle speed sensor cables occurred in an urban area in Japan. At the request of the police, DNA analysis was conducted to identify the animal species responsible. Swab samples collected from the damaged sections of the cables were subjected to PCR testing using mtDNA fragments. Sequencing analysis with universal primers (SCPH02500, SCPL02981) detected DNA from the Japanese marten (Martes melampus). A comprehensive examination that included morphological analysis of the cable damage and consideration of the ecological characteristics of the Japanese martens suggested that the damage was likely caused by this species. DNA analysis using mtDNA markers is a valuable tool for species identification in wildlife forensic veterinary investigations and serves as important scientific evidence in criminal cases involving animals. The findings from this case may contribute to future investigations in forensic veterinary science and ecological research and may also inform measures to prevent human–wildlife conflicts involving animals. Full article

(This article belongs to the Section Wildlife)

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21 pages, 8542 KiB

Open AccessArticle

An Efficient Algorithm for Small Livestock Object Detection in Unmanned Aerial Vehicle Imagery

by Wenbo Chen, Dongliang Wang and Xiaowei Xie

Animals 2025, 15(12), 1794; https://doi.org/10.3390/ani15121794 - 18 Jun 2025

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Abstract

Livestock population surveys are crucial for grassland management tasks such as health and epidemic prevention, grazing prohibition, rest grazing, and forage–livestock balance assessment. These tasks are integral to the modernization and upgrading of the livestock industry and the sustainable development of grasslands. Unmanned [...] Read more.

Livestock population surveys are crucial for grassland management tasks such as health and epidemic prevention, grazing prohibition, rest grazing, and forage–livestock balance assessment. These tasks are integral to the modernization and upgrading of the livestock industry and the sustainable development of grasslands. Unmanned aerial vehicles (UAVs) provide significant advantages in flexibility and maneuverability, making them ideal for livestock population surveys. However, grazing livestock in UAV images often appear small and densely packed, leading to identification errors. To address this challenge, we propose an efficient Livestock Network (LSNET) algorithm, a novel YOLOv7-based network. Our approach incorporates a low-level prediction head (P2) to detect small objects from shallow feature maps, while removing a deep-level prediction head (P5) to mitigate the effects of excessive down-sampling. To capture high-level semantic features, we introduce the Large Kernel Attentions Spatial Pyramid Pooling (LKASPP) module. In addition, we replaced the original CIoU with the WIoU v3 loss function. Furthermore, we developed a dataset of grazing livestock for deep learning using UAV images from the Prairie Chenbarhu Banner in Hulunbuir, Inner Mongolia. Our results demonstrate that the proposed module significantly improves the detection accuracy for small livestock objects, with the mean Average Precision (mAP) increasing by 1.47% compared to YOLOv7. Thus, this work offers a novel and practical solution for livestock detection in expansive farms. It overcomes the limitations of existing methods and contributes to more effective livestock management and advancements in agricultural technology. Full article

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

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2 pages, 127 KiB

Open AccessCorrection

Correction: Qin et al. Dietary Fermented Blueberry Pomace Supplementation Improves Small Intestinal Barrier Function and Modulates Cecal Microbiota in Aged Laying Hens. Animals 2024, 14, 2786

by Binghua Qin, Zhihua Li, Qian Zhu, Ting Chen, Wei Lan, Yadong Cui, Md. Abul Kalam Azad and Xiangfeng Kong

Animals 2025, 15(12), 1793; https://doi.org/10.3390/ani15121793 - 18 Jun 2025

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Abstract

Error in Table [...] Full article

(This article belongs to the Collection Use of Agricultural By-Products in Animal Feeding)

21 pages, 2449 KiB

Open AccessArticle

Toxic Effects of Acute Water Selenium Exposure on Litopenaeus vannamei: Survival, Physiological Responses, Transcriptome, and Intestinal Microbiota

by Xinghui Luo, Jian Chen, Asare Derrick, Gongyu Li, Hongming Wang, Zhihao Xue, Lili Shi and Shuang Zhang

Animals 2025, 15(12), 1792; https://doi.org/10.3390/ani15121792 - 18 Jun 2025

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Abstract

Excess selenium (Se) can cause a variety of toxic effects in aquatic animals. However, there is currently a lack of comprehensive studies about the toxicity effects of Se in culture water on shrimp. Based on the 96 h acute toxicity test, which confirmed [...] Read more.

Excess selenium (Se) can cause a variety of toxic effects in aquatic animals. However, there is currently a lack of comprehensive studies about the toxicity effects of Se in culture water on shrimp. Based on the 96 h acute toxicity test, which confirmed the 96 h LC₅₀ of Se (Na₂SeO₃) for Litopenaeus vannamei as 2.69 mg/L, L. vannamei (7.25 ± 0.05 g) were divided into three groups (named CON, S1 and S2) and exposed to Se at concentrations of 0, 0.0269 (0.01 × 96 h LC₅₀), and 0.269 (0.1 × 96 h LC₅₀) mg/L in the water for 72 h, respectively. The toxic effects of Se exposure on L. vannamei were evaluated based on histopathology, oxidative stress, immunity, apoptosis, transcriptional responses, and intestinal microbiota. Results demonstrated that Se exposure induced structural damage to the hepatopancreas of L. vannamei, including hepatocyte vacuolation and necrosis. Compared to the CON group, serum Caspase-3 activity significantly increased, while Bcl-2 activity markedly decreased in the S1 and S2 groups (p < 0.05). No significant differences in Bax activity were observed among groups (p > 0.05). ROS content, as well as activities of SOD, PO, GSH-PX, LYS, AKP, and ACP, exhibited an upward trend under Se exposure (p < 0.05). However, MDA levels showed no significant intergroup differences (p > 0.05). Hemocyte transcriptomic analysis revealed 2103 differentially expressed genes (DEGs) (1294 upregulated, 809 downregulated) in the S2 group compared to CON. GO enrichment indicated significant enrichment of DEGs in cellular processes, binding, and cell components. KEGG pathway analysis highlighted prominent enrichment in ribosome biogenesis in eukaryotes, lysosome, cell cycle, and pancreatic secretion pathways. Intestinal microbiota analysis showed that the Shannon, Simpson, and Pielou indices in the S2 group were significantly lower than those in the CON group (p < 0.05). The relative abundance of Vibrio and Acinetobacter increased significantly in the S2 group, while Enterococcus and Pseudomonas decreased markedly (p < 0.05). In conclusion, Se exposure triggered elevated immune enzyme activities, induced oxidative damage and apoptosis, transcriptional level metabolic disorders, and disrupted intestinal microbiota structure in L. vannamei. Full article

(This article belongs to the Special Issue Ecology of Aquatic Crustaceans: Crabs, Shrimps and Lobsters)

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