Animals

18 pages, 267 KB

Open AccessArticle

Generative AI in Veterinary Pathology: Feasibility of a GPT-Based Assistive Tool for Gross, Cytologic, and Histopathologic Assessment of Canine Cutaneous Neoplasms—A Pilot Study

by Evaristo Di Napoli, Luigi Emiliano Maria Zumbo, Davide De Biase, Giuseppe Piegari, Serenella Papparella, Valeria Russo and Orlando Paciello

Animals 2026, 16(13), 2070; https://doi.org/10.3390/ani16132070 (registering DOI) - 4 Jul 2026

Abstract

Canine cutaneous neoplasms are common and morphologically heterogeneous lesions whose diagnosis relies on integrating gross examination, cytology, and histopathology. This retrospective pilot study assessed the feasibility of a multimodal GPT-based large language model as an assistive, not autonomous, tool for standardized description, differential [...] Read more.

Canine cutaneous neoplasms are common and morphologically heterogeneous lesions whose diagnosis relies on integrating gross examination, cytology, and histopathology. This retrospective pilot study assessed the feasibility of a multimodal GPT-based large language model as an assistive, not autonomous, tool for standardized description, differential diagnosis generation, and classification support across this diagnostic workflow. Fifty-one histologically confirmed canine cutaneous tumors were retrospectively selected from the laboratory information system of the Veterinary Pathology Laboratory, University of Naples Federico II. For each case, de-identified gross photographs, digitized cytology, and representative histologic images were provided to the model using templated prompts. Model outputs were independently reviewed by two veterinary pathologists, who reached consensus on descriptive quality and diagnostic concordance with the histologic reference diagnosis. Final diagnostic outputs were classified as correct, partially correct, or incorrect. Strict accuracy was defined as the proportion of fully correct diagnoses, whereas broad accuracy combined correct and partially correct outputs considered diagnostically informative. Overall, the model achieved a strict diagnostic accuracy of 66.7% (34/51; 95% CI: 53.0–78.0) and a broad diagnostic accuracy of 90.2% (46/51; 95% CI: 79.0–95.7). Performance was highest in epithelial tumors and lower in mesenchymal and melanocytic tumors, in which the model more often identified broader diagnostic categories than specific histotypes. These findings suggest that GPT-based systems may support report standardization, descriptive consistency, and morphology-driven reasoning in veterinary pathology. However, reduced entity-level specificity, variable descriptive quality, and the risk of plausible but non-concordant outputs require strict human supervision and further validation before routine implementation. Full article

(This article belongs to the Special Issue Advances in Animal Oncology: Diagnostic Innovations, Therapeutic Approaches, and One Health Perspectives—2nd Edition)

19 pages, 3115 KB

Open AccessArticle

Multi-Omics Reveals Gut Microbiota Shifts and Hepatic Metabolic–Immune Alterations in “Short-Leg” Malformed Frog (Pelophylax nigromaculatus)

by Dan Zeng, Qin Qin, Ming Yang, Zi’ao Wang, Jianguo Xiang, Xiaoqing Wang and Yazhou Hu

Animals 2026, 16(13), 2069; https://doi.org/10.3390/ani16132069 (registering DOI) - 4 Jul 2026

Abstract

Amphibian malformation syndromes significantly impact both conservation efforts and aquaculture, yet their underlying systemic pathophysiological mechanisms remain poorly characterized. This study comprehensively examines the multi-level pathological processes associated with the “short-leg” malformation syndrome in the black-spotted frog (Pelophylax nigromaculatus) using an [...] Read more.

Amphibian malformation syndromes significantly impact both conservation efforts and aquaculture, yet their underlying systemic pathophysiological mechanisms remain poorly characterized. This study comprehensively examines the multi-level pathological processes associated with the “short-leg” malformation syndrome in the black-spotted frog (Pelophylax nigromaculatus) using an integrated methodology, encompassing morphological, histopathological, gut microbiome, and hepatic transcriptomic analyses. Affected frogs demonstrated shortened limbs, impaired motor function, and a distinctive metabolic phenotype, including increased body weight despite a shorter body length, accumulation of visceral fat, and shortened intestines. Gut microbiota analysis identified significant compositional shifts, characterized by a decreased Firmicutes-to-Bacteroidota ratio, expansion of pro-inflammatory Proteobacteria, and reduction in beneficial Actinobacteriota, suggesting microbial niche restructuring that likely promotes metabolic and inflammatory disorders. Hepatic transcriptome profiling revealed 2617 differentially expressed genes, demonstrating a clear molecular dichotomy with concurrent up-regulation of immune-related pathways (e.g., neutrophil extracellular trap formation, complement cascades, and inflammatory signaling) and broad suppression of metabolic pathways (e.g., lipid oxidation, nutrient absorption, and PPAR and renin–angiotensin systems). This integrated analysis illustrates that the malformation syndrome represents a systemic pathophysiological state involving dysfunction of the gut–liver axis, characterized by the coexistence of gut microbiota alterations, hepatic metabolic suppression, and immune activation. These findings provide a framework for understanding amphibian malformations and suggest potential strategies to improve health outcomes in aquaculture. Full article

(This article belongs to the Section Aquatic Animals)

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15 pages, 7063 KB

Open AccessArticle

Mechanisms Underlying the Impact of Feed-to-Gain Ratio Differences on Nutrient Metabolism in Simmental and Simmental × Hereford Crossbred Cattle Fed a Low-Energy Diet

by Yi Wu, Hao Chen, Danling Zhang, Lina Wang, Qi Liu, Chunjie Wang, Aorigele Chen and Hairong Wang

Animals 2026, 16(13), 2068; https://doi.org/10.3390/ani16132068 (registering DOI) - 4 Jul 2026

Abstract

To systematically investigate the metabolic and microbial differences in feed efficiency between Hereford × Simmental (H × S) crossbred and purebred Simmental cattle (S × S), 20 steers (initial weight = 268.80 ± 30.88 kg; initial age = 273.00 ± 14.09 days) were [...] Read more.

To systematically investigate the metabolic and microbial differences in feed efficiency between Hereford × Simmental (H × S) crossbred and purebred Simmental cattle (S × S), 20 steers (initial weight = 268.80 ± 30.88 kg; initial age = 273.00 ± 14.09 days) were used, which were housed in mixed pens and fed a low-energy diet (0.75 Mcal Neg/kg DM). This study revealed that the level of Hypoglycin B was related to a reduced T3, which could affect growth performance. Metabolomics analysis identified Hypoglycin B, lysoPC, and DHAP as key discriminative metabolites. Pathway enrichment analysis revealed that these metabolites were primarily mapped to glycine, serine, and threonine metabolism; alanine, aspartate, and glutamate metabolism; and pantothenate and CoA biosynthesis. In our study, the relative abundances of Succinivibrio and Clostridium_sensu_stricto_6 were significantly greater in the H × S group than in the S × S group; it may have stronger energy extraction abilities and matches their lower F/G. Romboutsia was positively correlated with Hypoglycin B and negatively correlated with LysoPC (20:2 (11Z,14Z)/0:0), which might be related to changes in blood metabolites. Nevertheless, these explanations remain correlational and require functional validation through targeted interventions. Full article

(This article belongs to the Section Animal Nutrition)

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17 pages, 2060 KB

Open AccessArticle

Antennal Transcriptome Profiling Reveals Gustatory Receptors Associated with Pollen Foraging Preferences in Apis mellifera

by Qiyan Su, Yu Zhang, Chang Song, Lina Guo and Yuan Guo

Animals 2026, 16(13), 2067; https://doi.org/10.3390/ani16132067 (registering DOI) - 4 Jul 2026

Abstract

Gustatory perception in honeybees is a key determinant of foraging decisions and pollen source selection. However, the molecular mechanisms underlying this sensory discrimination remain poorly understood. To investigate these mechanisms during the collection of pollen from different floral sources, this study utilized antennae [...] Read more.

Gustatory perception in honeybees is a key determinant of foraging decisions and pollen source selection. However, the molecular mechanisms underlying this sensory discrimination remain poorly understood. To investigate these mechanisms during the collection of pollen from different floral sources, this study utilized antennae from worker bees foraging on pear and rapeseed pollen, and non-pollen-foraging workers as controls. Illumina high-throughput transcriptome sequencing was employed to identify differentially expressed genes (DEGs), perform functional annotation, and characterize gustatory receptor (GR) genes. Compared with the control group, 583 DEGs and 516 DEGs were identified in pear-pollen and rapeseed-pollen foragers, respectively, whereas only 73 DEGs were detected between the two pollen-foraging groups. Several DEGs were associated with chemosensory perception, signal transduction, energy metabolism, and immune responses. Notably, genes involved in membrane-associated signaling and stimulus response exhibited differential expression patterns among foraging groups, suggesting adaptive molecular responses to distinct floral resources. Gene Ontology (GO) analysis indicated that DEGs were primarily associated with cellular processes, membrane components, and binding functions. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment highlighted significant involvement in phagosome, phosphatidylinositol signaling system, oxidative phosphorylation, and extracellular matrix–receptor interaction. Notably, seven GR-related genes were identified in the antennal transcriptome, including five known GR genes and two novel candidates, all with complete open reading frames. Four of these genes featured the canonical seven-transmembrane domain structure of insect GRs. Phylogenetic analysis, in addition to the known sugar receptors AmelGR43a, AmelGR64f, and AmelGR64f-X1, based on GRs from Apis mellifera and Drosophila melanogaster suggested that AmelGR28b, AmelGR10, AmelGR12, and AmelGR13 may belong to the bitter taste receptor family. Quantitative real-time PCR (qRT-PCR) validation demonstrated that the expression patterns of the selected seven DEGs were consistent with the RNA-seq results. This study reveals differential expression patterns and potential functional divergence of gustatory receptor genes in Apis mellifera during pollen collection from different floral sources. It provides important molecular evidence for understanding how honeybees accurately recognize and preferentially forage specific pollen sources via gustatory perception, and offers valuable theoretical and practical insights for honeybee behavioral ecology and crop pollination management. Full article

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

14 pages, 333 KB

Open AccessArticle

Association Between Dietary Regimen and Renal Function Parameters in African Pygmy Hedgehogs (Atelerix albiventris)

by Kristina Spariosu, Ana Pešić, Ksenija Nešić, Diana Brozić, Jelena Francuski Andrić, Branislav Vejnović and Miloš Vučićević

Animals 2026, 16(13), 2066; https://doi.org/10.3390/ani16132066 (registering DOI) - 4 Jul 2026

Abstract

African pygmy hedgehogs (APHs) are increasingly kept as companion animals, yet evidence-based nutritional recommendations for this species remain limited. Commercial cat or kitten diets are still commonly used by owners and veterinarians in practice, despite being formulated exclusively for domestic cats rather than [...] Read more.

African pygmy hedgehogs (APHs) are increasingly kept as companion animals, yet evidence-based nutritional recommendations for this species remain limited. Commercial cat or kitten diets are still commonly used by owners and veterinarians in practice, despite being formulated exclusively for domestic cats rather than for hedgehogs with omnivorous–insectivorous feeding habits. This cross-sectional study evaluated the association between dietary regimen and serum biochemical parameters associated with renal function in APHs. Blood samples were collected from 19 client-owned APHs aged 12–68 months: 10 hedgehogs fed a commercial kitten diet and 9 fed a commercial APH-specific diet. Serum concentrations of blood urea nitrogen, creatinine, phosphorus, and calcium were measured, and the nutritional composition of the two diets was compared using manufacturer declarations and targeted laboratory analyses. Hedgehogs fed the commercial kitten diet had significantly higher serum blood urea nitrogen (p = 0.0133), creatinine (p = 0.0279), and phosphorus (p = 0.0279) concentrations than those fed the APH-specific diet, whereas serum calcium concentrations did not differ significantly (p = 0.3846). These differences occurred despite similar dietary phosphorus content, while the commercial kitten diet had higher declared fat and lower fiber content. The findings suggest that dietary regimen may be associated with alterations in renal biochemical profiles in APHs and support the use of species-appropriate diets in clinical practice. Full article

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

26 pages, 7600 KB

Open AccessArticle

Spatial–Temporal Dynamics and Co-Occurrence Networks of Zooplankton Community Structure in a Large Shallow Reservoir Under the Background of Ecological Restoration

by Ting Yuan, Jiangqianhui Qi, Mantang Xiong, Geng Huang, Kui Zhang, Lianfeng Zhou and Feng Chen

Animals 2026, 16(13), 2065; https://doi.org/10.3390/ani16132065 (registering DOI) - 4 Jul 2026

Abstract

Zooplankton are sensitive indicators of ecological change, but their community reorganization under early-phase restoration remains unclear. This study aims to reveal how their diversity, standing stock, and co-occurrence networks respond to early-phase restoration and identifies key environmental drivers, offering novel insights into restoration–plankton [...] Read more.

Zooplankton are sensitive indicators of ecological change, but their community reorganization under early-phase restoration remains unclear. This study aims to reveal how their diversity, standing stock, and co-occurrence networks respond to early-phase restoration and identifies key environmental drivers, offering novel insights into restoration–plankton interplay. Shilianghe Reservoir, as the largest reservoir in Jiangsu Province, represents a typical case of restoration in the Huai River Basin that offers transferable insights for numerous eutrophic reservoirs. The study comprised eight surveys representing two annual cycles (July 2021–May 2022 and April 2023–January 2024). The 2023 sampling cycle showed lower zooplankton density and biomass but higher α-diversity than the 2021 sampling cycle. Co-occurrence network density and the size of the largest connected component decreased in spring and summer of the 2023 sampling cycle, suggesting community reorganization. Inflow tributaries supported the highest species richness and standing stock but showed looser interspecific associations. PERMANOVA analysis indicated that season explained the largest proportion of community variation, followed by sampling year and region. Environmental variables including DO, WT, Cond and TN significantly correlated with zooplankton community structure. Zooplankton community reorganization was characterized by reduced standing stock, increased diversity, and altered co-occurrence patterns, providing a mechanistic framework for using zooplankton as bioindicators in reservoir restoration assessments. Full article

15 pages, 3987 KB

Open AccessArticle

Monitoring Animal Poisoning in Central Italy: Long-Term Trends and Spatial Distribution in the Abruzzo Region

by Antonio Petrini, Alice Menozzi, Carmine Merola, Giampiero Scortichini, Daria Di Sabatino, Luca Brugnola, Stefania Salucci, Elga Ersilia Tieri, Daniela Averaimo, Antonio Cocco, Giorgio Saluti, Giuseppe Gatti, Rachele Rocchi, Davide De Tiberis, Maria Laura Marchetti, Daniel Buldain, Pedro Zeinsteger and Pietro Badagliacca

Animals 2026, 16(13), 2064; https://doi.org/10.3390/ani16132064 (registering DOI) - 4 Jul 2026

Abstract

Both domestic and wild animals are frequently affected by accidental, intentional, or illegal poisoning globally, but data from some regions, including Italy, remain scarce. This study aimed to characterize animal poisoning events in a central Italian region over the period 2011–2020. A total [...] Read more.

Both domestic and wild animals are frequently affected by accidental, intentional, or illegal poisoning globally, but data from some regions, including Italy, remain scarce. This study aimed to characterize animal poisoning events in a central Italian region over the period 2011–2020. A total of 2722 suspected cases were analyzed, of which 51.3% were confirmed positive for at least one poison. Among the 1397 confirmed cases, 863 animal carcasses and 534 poisoned baits were recorded, corresponding to 762 distinct poisoning events. The most frequently involved toxicants were metaldehyde, carbamates, and organophosphorus pesticides, followed by anticoagulant rodenticides and other compounds. Spatial analysis revealed distinct patterns in the use of toxicants. Sub-regional areas encompassing national and regional parks were primarily associated with agricultural pesticides, whereas urban areas were dominated by metaldehyde and anticoagulant rodenticides. These differences suggest that, in addition to their illegal nature, poisoning events may reflect underlying socio-environmental conflicts between humans and animals. The results highlight the value of an interdisciplinary approach to mitigate poisoning, involving coordinated efforts among scientific, veterinary, enforcement, and public stakeholders. Such strategies could reduce both the frequency and ecological impact of this illegal practice. Full article

(This article belongs to the Special Issue Xenobiotic Exposure and Toxicological Effects: A One Health Approach)

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15 pages, 1694 KB

Open AccessArticle

Rapid Avian Diversity Recovery Following Photovoltaic Module Removal: Rebounds in Larger Waterbirds Composition and Habitat Restoration in Lake Littoral Areas

by Lei Cheng, Bingguo Dai, Zhenhua Wei and Shuyue Cheng

Animals 2026, 16(13), 2063; https://doi.org/10.3390/ani16132063 (registering DOI) - 4 Jul 2026

Abstract

In the context of accelerated progress in clean energy development, an increasing number of regions are utilizing photovoltaic modules (PVMs) for the purpose of energy production. Numerous studies have demonstrated that the installation of these PVMs can exert both positive and negative effects [...] Read more.

In the context of accelerated progress in clean energy development, an increasing number of regions are utilizing photovoltaic modules (PVMs) for the purpose of energy production. Numerous studies have demonstrated that the installation of these PVMs can exert both positive and negative effects on local ecosystems in different regions. However, there is a paucity of studies that have explored the ecological recovery mechanisms following PVM removal from another perspective, which may hinder a comprehensive understanding of PVM ecological impacts and impede the formulation and implementation of ecological management policies. The PVMs installed on Jiaogang Lake, which were to be removed due to policy adjustments, provided a valuable opportunity to address this issue. The taxonomic and functional diversity of waterbird communities in Jiaogang Lake was assessed and compared before and after the removal of PVMs. The results demonstrated that following the removal, there was a significant increase in both waterbird species richness and functional diversity, whilst the functional nestedness patterns between waterbird communities became more pronounced, implying the restoration of lake littoral areas. Subsequent analysis of individual waterbird functional traits revealed a significant trend towards larger body sizes in waterbirds following the removal of PVMs. Although temporal changes in certain traits did not attain statistical significance, the consistent trends observed in numerous traits led to this extrapolation with confidence. The present study thus aims to encourage decisive decision-making, particularly in short-term macroecological research in the local-scale region, avoiding delays in ecological management measures due to an obsession with perfect statistical data and analysis outcomes. Full article

(This article belongs to the Section Birds)

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17 pages, 1327 KB

Open AccessArticle

Longitudinal Multimodal Monitoring of Eight Captive Beluga Whale (Delphinapterus leucas) Pregnancies over a 25-Year Period

by Takashi Kamio, Wataru Ohtomo, Yuichiro Akune, Masanori Kurita and Yasuo Inoshima

Animals 2026, 16(13), 2062; https://doi.org/10.3390/ani16132062 - 3 Jul 2026

Abstract

The accurate prediction of parturition in managed beluga whales (Delphinapterus leucas) is fundamental for optimizing maternal and neonatal care; however, reliable predictive indicators remain limited. Here, eight pregnancies (five live births and three adverse pregnancy outcomes) monitored over 25 years at [...] Read more.

The accurate prediction of parturition in managed beluga whales (Delphinapterus leucas) is fundamental for optimizing maternal and neonatal care; however, reliable predictive indicators remain limited. Here, eight pregnancies (five live births and three adverse pregnancy outcomes) monitored over 25 years at a single facility were retrospectively analyzed. The rectal temperatures, serum progesterone concentrations, gestation lengths, food intake, behavioral changes, and fetal heart rates of the whales were evaluated, particularly prepartum. Five successful pregnancies exhibited consistent seasonal timing and reproducible endocrine and physiological trajectories. The mean gestation length was 466 ± 8.4 days. The rectal temperatures of dams that delivered live offspring decreased by 1.6 ± 0.5 °C approximately 1.3 ± 0.5 days before parturition. In successful pregnancies, serum progesterone concentrations declined prepartum but typically remained detectable until parturition. In contrast, a concentration of approximately 1 ng/mL prior to parturition was observed in the pregnancy that resulted in stillbirth. Adverse pregnancy outcomes were associated with deviations from the patterns observed in successful pregnancies, including abnormal gestation length, notably reduced progesterone concentrations, altered fetal heart rate trajectories, and ultrasonographic evidence of fetal cranial asymmetry. These findings highlight the importance of integrated multimodal monitoring in predicting parturition and identifying abnormal pregnancy progression in managed beluga whales. Full article

(This article belongs to the Special Issue Advances in the Reproduction of Wild and Exotic Animals)

17 pages, 2536 KB

Open AccessReview

Decapod Crustaceans in Animal Welfare Law: Fragmentation, Gaps, and Emerging Models in Europe and Oceania

by Lorenzo Fruscella, Diego Antonio Sicuso, Daria Vitale and Annamaria Passantino

Animals 2026, 16(13), 2061; https://doi.org/10.3390/ani16132061 - 3 Jul 2026

Abstract

In recent decades, the exponential growth of the global seafood industry has intensified the exploitation of aquatic invertebrates, particularly decapod crustaceans, whose commercial value continues to rise alongside consumer demand. This expansion has coincided with significant scientific advances recognizing these animals as sentient [...] Read more.

In recent decades, the exponential growth of the global seafood industry has intensified the exploitation of aquatic invertebrates, particularly decapod crustaceans, whose commercial value continues to rise alongside consumer demand. This expansion has coincided with significant scientific advances recognizing these animals as sentient beings capable of experiencing pain and exhibiting complex behavioral responses to noxious stimuli. Nevertheless, legal systems have been slow integrate such evidence, resulting in a persistent misalignment between scientific knowledge and regulatory practice. This article critically examines the legal status of decapod crustaceans within animal welfare law, underscoring a pronounced disjunction between well-established scientific evidence of their sentience and the limited protections they receive. Despite extensive research supporting their capacity for suffering, the prevailing legal paradigm continues to construe these animals predominantly as commodities. Through a comparative analysis of regulatory regimes across the European Union, Italy, and selected extra-European jurisdictions, including UK, Australia, and New Zealand, the study identifies a significant regulatory lacuna alongside considerable legislative fragmentation. It scrutinizes a spectrum of normative models, ranging from indeterminate, principle-based provisions to more exacting and technically prescriptive standards. The article finally advocates for a harmonized, science-driven regulatory framework to ensure coherent and effective welfare protections for decapod crustaceans within the global food industry. Full article

(This article belongs to the Section Animal Welfare)

25 pages, 3591 KB

Open AccessArticle

Infrared Thermography for Simultaneous Detection of Limb Pathology and Subclinical Mastitis in Dairy Cows

by Juozas Žemaitis, Ugnė Spancernienė, Vaida Jokubauskienė, Ignas Šilinskas, Kristina Musayeva, Rasa Želvytė, Judita Žymantienė, Antanas Sederevičius, Joris Vėžys, Vytautas Jūrėnas, Algimantas Bubulis, Sigitas Japertas, Vytautas Ostaševičius and Vaidas Oberauskas

Animals 2026, 16(13), 2060; https://doi.org/10.3390/ani16132060 - 3 Jul 2026

Abstract

This study evaluated infrared thermography (IRT) as a dual-purpose, non-invasive tool for detecting limb pathology and subclinical mastitis in dairy cows. Surface temperatures were measured at the coffin, pastern, and fetlock regions of all limbs and at the teat surface of all udder [...] Read more.

This study evaluated infrared thermography (IRT) as a dual-purpose, non-invasive tool for detecting limb pathology and subclinical mastitis in dairy cows. Surface temperatures were measured at the coffin, pastern, and fetlock regions of all limbs and at the teat surface of all udder quarters. Limb health status was determined by orthopedic clinical examination, and mastitis status was assessed using the California Mastitis Test (CMT). Hindlimbs were significantly warmer than forelimbs in healthy cows (p < 0.001), with confirmed bilateral symmetry (r = 0.524–0.569). Limbs with pathology showed significantly higher temperatures at all anatomical sites, with particularly large effect sizes in the hindlimbs (r around 0.80). Additional lesion-specific analyses demonstrated that both infectious and claw horn lesions exhibited significantly higher temperatures than healthy limbs. In the hindlimbs, infectious lesions showed higher thermographic values than claw horn lesions. CMT-positive quarters showed teat temperatures of 30.0–30.2 °C compared with 25.2–25.8 °C in CMT-negative quarters (p < 0.001). Receiver operating characteristic (ROC) analysis demonstrated excellent diagnostic performance for teat surface temperature in identifying CMT-positive quarters, with an area under the ROC curve (AUC) of 0.956. The optimal threshold was 29.5 °C, corresponding to a sensitivity of 0.992 and a specificity of 0.838. Each 1 °C increase in teat temperature increased the odds of CMT positivity by 5.3-fold (p < 0.001). A practical three-zone thermographic framework was established: <27 °C (healthy), 27–29.5 °C (monitoring recommended), and ≥29.5 °C (mastitis likely). Teat temperature was not associated with composite milk somatic cell count, but hindlimb pathology was independently associated with elevated teat temperatures (partial r = 0.23–0.26, p ≤ 0.001). These findings demonstrate that IRT has strong potential as an integrated screening tool for dairy cow limb health and udder health monitoring. Full article

(This article belongs to the Topic Application of Animal Models: From Physiology to Pathology, 2nd Edition)

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16 pages, 633 KB

Open AccessArticle

Effect of Zinc Hydroxychloride and Copper Hydroxychloride in Compound Feed on Rearing Results and Carcass Characteristics of Broiler Chickens

by Sabina Kaim, Dorota Banaszewska and Barbara Biesiada-Drzazga

Animals 2026, 16(13), 2059; https://doi.org/10.3390/ani16132059 - 3 Jul 2026

Abstract

The aim of the study was to compare the production results, slaughter value, and physicochemical and biochemical parameters of the muscles of Ross 308 broiler chickens. The study included 225 individuals divided into three groups of 75 birds each. Each group was subdivided [...] Read more.

The aim of the study was to compare the production results, slaughter value, and physicochemical and biochemical parameters of the muscles of Ross 308 broiler chickens. The study included 225 individuals divided into three groups of 75 birds each. Each group was subdivided into three subgroups of 25 birds each (replicates). The control group received zinc (Zn) in the form of zinc oxide (ZnO) and copper (Cu) in the form of copper (II) sulfate pentahydrate (CuSO₄·5H₂O). Experimental group I received the microelements zinc and copper exclusively in the form of zinc hydroxychloride and dicopper chloride trihydroxide. Experimental group II received zinc and copper in both inorganic and hydroxychloride forms. The obtained results were processed using statistical analysis using the STATISTICA 13.0 [2016] program. The significance of differences between groups was inferred based on Tukey’s test. Chickens from the experimental groups were characterized by higher body weight (p ≤ 0.05) and eviscerated carcass weight (p ≤ 0.05) and better feed utilization (p ≤ 0.05). Moreover, compared with the control group, the carcasses of chickens from the experimental groups were characterized by a lower weight of breast muscles (p ≤ 0.05) and skin with subcutaneous fat, a similar weight of leg muscles, and a significantly greater weight of the remaining carcass components (p ≤ 0.01). The use of Zn and Cu in the form of hydroxychlorides in broiler chicken nutrition increased the fat content in the muscles (p ≤ 0.05), the concentrations of Zn and Cu in the liver, and significantly improved the tibial bone strength of broiler chickens. The application of Zn and Cu in the form of hydroxychlorides in the diets (experimental groups I and II) enhanced lipid and protein oxidation processes and the overall antioxidant capacity in the breast muscle. To conclude, dietary supplementation with zinc and copper hydroxychlorides in broilers promotes growth performance, bone strength, and mineral bioavailability, though it reduces breast muscle yield and triggers oxidative stress in these tissues. Full article

(This article belongs to the Special Issue Feed Additives in Poultry Industry)

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49 pages, 7831 KB

Open AccessReview

Recent Advances in Vision-Based Beef Cattle Body Measurement Technologies

by Xiaofan Deng, Fuli Zhang, Gang Jin, Liangyu Cui, Dongxu Zhang and Fa Zhang

Animals 2026, 16(13), 2058; https://doi.org/10.3390/ani16132058 - 3 Jul 2026

Abstract

Accurate beef cattle body measurement data are crucial for growth assessment, phenotypic analysis, breeding management, and precision livestock farming. Traditional manual measurements are labor-intensive, time-consuming, and likely to cause stress in animals, making it difficult to meet the demands of large-scale livestock farming. [...] Read more.

Accurate beef cattle body measurement data are crucial for growth assessment, phenotypic analysis, breeding management, and precision livestock farming. Traditional manual measurements are labor-intensive, time-consuming, and likely to cause stress in animals, making it difficult to meet the demands of large-scale livestock farming. This paper employs a structured systematic literature review method, in accordance with the PRISMA 2020 guidelines, to summarize research progress in vision-based beef cattle body measurement. This paper focuses on reviewing technical approaches such as 2D image-based measurement, 3D measurement using RGB-D and LiDAR, and multi-view fusion. It analyzes key technologies including image segmentation, keypoint detection, point cloud processing, 3D reconstruction, and geometric calculations, and compares the advantages and disadvantages of different methods in terms of measurement accuracy, robustness, cost, and farm applicability. The results indicate that 2D image-based methods are low-cost and flexible to deploy but have limited expressiveness for 3D body measurement parameters; RGB-D and LiDAR methods can provide spatial information but are affected by point cloud noise, occlusion, equipment costs, and data processing complexity; multi-view fusion can improve the completeness of body surface information but places high demands on calibration, registration, and system integration. Current research still faces challenges such as a lack of public datasets, inconsistent annotation standards, uncertainty regarding ground truth, insufficient cross-ranch generalization validation, and limited practical applications. Future research should focus on developing standardized datasets, conducting cross-scenario validation, advancing multimodal perception, creating lightweight models, and applying edge computing to drive the evolution of visual body measurement toward continuous monitoring and intelligent decision-making. Full article

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

27 pages, 8157 KB

Open AccessArticle

An Enhanced Particle Swarm Optimized RBF Model for Precise Fish Population Estimation in Cage Farming

by Gang Yang, Xuelei Wang, Junping Wang, Weiliang Shen, Hongsheng Yang, Qingfei Li and Chenggang Lin

Animals 2026, 16(13), 2057; https://doi.org/10.3390/ani16132057 - 3 Jul 2026

Abstract

In cage aquaculture, precise estimation of fish biomass is critically important for determining appropriate feeding strategies and evaluating production capacity. Currently, prevailing fish counting approaches heavily rely on acoustic or optical technologies. However, the accuracy and reliability of the obtained data are largely [...] Read more.

In cage aquaculture, precise estimation of fish biomass is critically important for determining appropriate feeding strategies and evaluating production capacity. Currently, prevailing fish counting approaches heavily rely on acoustic or optical technologies. However, the accuracy and reliability of the obtained data are largely compromised by factors such as fish occlusion and water turbidity in practical cage farming environments. To address this limitation, this study proposed a novel method for estimating fish population size deduced from dynamic feeding information, based on the model integrated environmental and biological factors, feed intake and biomass. A 10-week feeding experiment was carried out to collect multidimensional data including feed intake, growth parameters, and environmental variables to construct a dataset correlating feeding amount with primary influential factors. Herein a bioenergetics-informed radial basis function neural network, optimized via particle swarm optimization (BE-PSO-RBF), was developed based on those empirical data. Model validation using 47 independent test samples showed that the hybrid model achieved a mean absolute error (MAE) of 26.82, a root mean square error (RMSE) of 35.62, and a mean absolute percentage error (MAPE) of 4.14%, confirming its robust generalization performance. These findings suggest that feed-intake-based population estimation may provide a feasible complementary approach for fish population assessment under cage farming conditions similar to those investigated in this study. Full article

(This article belongs to the Section Aquatic Animals)

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9 pages, 214 KB

Open AccessPerspective

Informal Treatment Practices in Ornamental Aquaria: An Overlooked Interface Between Aquatic Animal Health, Antimicrobial Stewardship, and One Health

by Marco Dettori

Animals 2026, 16(13), 2056; https://doi.org/10.3390/ani16132056 - 3 Jul 2026

Abstract

Ornamental aquarium keeping collectively involves millions of freshwater, marine, and reef systems in which fish, corals, invertebrates, biofilters, microbial communities, and human husbandry practices are closely interconnected. In these domestic aquatic animal systems, preventive and curative treatments may include antimicrobials, antiparasitics, antiseptics, oxidizing [...] Read more.

Ornamental aquarium keeping collectively involves millions of freshwater, marine, and reef systems in which fish, corals, invertebrates, biofilters, microbial communities, and human husbandry practices are closely interconnected. In these domestic aquatic animal systems, preventive and curative treatments may include antimicrobials, antiparasitics, antiseptics, oxidizing agents, copper-based products, dips, and commercial formulations targeting microbial proliferations or visible system deterioration. Many interventions occur without veterinary diagnosis, microbiological confirmation, standardized dosing, active-ingredient transparency, or post-treatment monitoring. This raises concerns for aquatic animal health and welfare, as whole-system treatments may affect not only the intended pathogen or pest but also non-target organisms, biofilter communities, animal-associated microbiota, and water quality stability. Digital communities and online platforms can rapidly circulate empirical treatment protocols, although they may also provide opportunities for stewardship education and improved husbandry guidance. Current evidence does not support interpreting ornamental aquaria as major independent drivers of antimicrobial resistance. The more defensible concern is stewardship: biologically active compounds may be used repeatedly and empirically in animal systems without diagnosis, professional guidance, or systematic monitoring. This Perspective argues that ornamental aquaria should be recognized as an overlooked interface between aquatic animal health, welfare, antimicrobial stewardship, and One Health. It proposes a research and communication agenda focused on treatment transparency, diagnosis, prevention, biofilter protection, and responsible care practices. Full article

(This article belongs to the Section Aquatic Animals)

16 pages, 14909 KB

Open AccessArticle

Reproductive Traits Revealing the Invasion and Coexistence of Two Tilapia Species in the Jinghong Reservoir of the Lower Lancang River, Southwest China

by Ziheng Hu, Liwen Dong, Ke Li, Dongdong Zhai, Yuanyuan Chen, Hongyan Liu, Fei Xiong, Xinbin Duan and Mingdian Liu

Animals 2026, 16(13), 2055; https://doi.org/10.3390/ani16132055 - 3 Jul 2026

Abstract

Coptodon zillii and Oreochromis niloticus are dominant invasive fish species that have successfully established and coexisted in the Jinghong Reservoir of the lower Lancang River, Southwest China. To explore the mechanisms underlying their invasion and coexistence from the perspective of reproductive traits, we [...] Read more.

Coptodon zillii and Oreochromis niloticus are dominant invasive fish species that have successfully established and coexisted in the Jinghong Reservoir of the lower Lancang River, Southwest China. To explore the mechanisms underlying their invasion and coexistence from the perspective of reproductive traits, we conducted monthly sampling from January to December 2025 to compare key reproductive parameters, including size at first sexual maturity, sex ratio, breeding season, spawning pattern and fecundity. The results showed that the predicted sizes at first sexual maturity for females and males of C. zillii (83.4 mm and 81.7 mm, respectively) were significantly smaller than those of O. niloticus (127.7 mm and 125.8 mm, respectively). Both species exhibited early sexual maturation. The sex ratio of C. zillii was approximately 1:1, whereas that of O. niloticus was significantly female-biased. The breeding season of C. zillii lasted from April to September, peaking in May and June, while O. niloticus spawned from May to November, with a peak in August, showing a staggered temporal distribution. Both species were batch spawners; however, O. niloticus had significantly larger egg diameters than C. zillii. Absolute fecundity, length-relative fecundity and weight-relative fecundity were significantly higher in C. zillii than in O. niloticus. Moreover, the absolute fecundity of C. zillii showed stronger correlations with body length, body weight, net weight and gonadal weight. C. zillii adopted a more r-selected strategy, while O. niloticus exhibited a more K-selected strategy. The two species displayed clear divergence in their reproductive strategies, including size at maturity, breeding season, egg diameter and fecundity, which reduced interspecific competition and promoted niche separation and coexistence in the Jinghong Reservoir. Full article

(This article belongs to the Special Issue Fish Reproductive Biology and Embryogenesis)

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31 pages, 5859 KB

Open AccessSystematic Review

Bacillus subtilis Supplementation in Weanling Piglets: A Systematic Review of Growth, Gut Health, and Microbiota Modulation

by Charlotte Ludorf, Carley Richardson and Kwangwook Kim

Animals 2026, 16(13), 2054; https://doi.org/10.3390/ani16132054 - 3 Jul 2026

Abstract

Bacillus subtilis has been widely investigated as a probiotic feed additive for weanling piglets due to its potential to improve growth performance, gut health, and disease resilience during the post-weaning period. This systematic review evaluated the effects of Bacillus subtilis supplementation in pigs [...] Read more.

Bacillus subtilis has been widely investigated as a probiotic feed additive for weanling piglets due to its potential to improve growth performance, gut health, and disease resilience during the post-weaning period. This systematic review evaluated the effects of Bacillus subtilis supplementation in pigs following PRISMA guidelines and the PICOS framework. A total of 619 records published between 2000 and 2025 were identified through PubMed, Scopus, and AGRICOLA databases, of which 29 studies met the inclusion criteria for qualitative synthesis. Overall, Bacillus subtilis supplementation frequently improved average daily gain, body weight, feed efficiency, and reduced diarrhea incidence, particularly under enteric challenge conditions. Many studies also reported beneficial effects on intestinal morphology, immune regulation, intestinal barrier-related gene expression, nutrient digestibility, and modulation of gut microbiota toward bacterial communities associated with intestinal health. However, responses varied depending on Bacillus subtilis strain, dosage, diet composition, and experimental design. In contrast, effects on biochemical and oxidative stress indicators were less consistent across studies. Collectively, the findings support the potential role of Bacillus subtilis as a functional probiotic in swine nutrition while highlighting the need for standardized long-term studies to better define optimal strains, supplementation strategies, and mechanisms of action. Full article

(This article belongs to the Special Issue Strategies to Improve Gut Health and Immunity in Monogastric Animals)

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22 pages, 7396 KB

Open AccessArticle

Integrated Lipidomic and Amino Acid Metabolomic Analyses Reveal Muscle Metabolic Differences in Tibetan Sheep Under Grazing and House-Feeding Systems

by Pengfei Zhao, Jianming Ren, Lan Zhang, Shiyu Tao, Chunyang Li, Ying Ma and Xiong Ma

Animals 2026, 16(13), 2053; https://doi.org/10.3390/ani16132053 - 3 Jul 2026

Abstract

Production system may affect meat quality and muscle metabolic characteristics in Tibetan sheep. In this study, the biceps femoris muscles of twelve 3-year-old Tibetan sheep with similar body weights were used as experimental materials during a 6-month experimental period. The housed group (n [...] Read more.

Production system may affect meat quality and muscle metabolic characteristics in Tibetan sheep. In this study, the biceps femoris muscles of twelve 3-year-old Tibetan sheep with similar body weights were used as experimental materials during a 6-month experimental period. The housed group (n = 6) was defined as the control group (C group), whereas the grazing group (n = 6) was defined as the L group. Meat quality measurement, nutritional composition analysis, untargeted lipidomics, and amino acid metabolomics (AAM) were integrated to investigate the effects of contrasting grazing and house-feeding production systems on meat quality and metabolic characteristics in Tibetan sheep. The results showed that cooking loss and drip loss were significantly decreased, whereas water-holding capacity (WHC) was significantly increased in the L group. However, shear force was also increased, indicating that grazing and house-feeding systems were associated with differences in muscle WHC and shear force. The L group exhibited significant alterations in lipid composition and increased concentrations of several n-3 polyunsaturated fatty acids and increased levels of omega-3 polyunsaturated fatty acids (n-3 PUFAs), including α-linolenic acid (ALA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA), suggesting that grazing and house-feeding systems were associated with differences in the lipid nutritional profile of muscle. Lipidomic analysis showed that the differential lipids were mainly enriched in triacylglycerols (TGs), phosphatidylethanolamines (PEs), and phosphatidylcholines (PCs), and several PUFA-containing TGs and membrane lipid molecules were closely associated with meat quality traits. AAM analysis showed that branched-chain amino acids (BCAAs), including L-leucine and L-valine, as well as N,N-dimethylglycine, were upregulated in the L group, whereas kynurenine and 1-methyl-L-histidine were downregulated. These findings suggest that BCAA metabolism and tryptophan–kynurenine metabolism were associated with metabolic differences observed between production systems in muscle metabolic adaptation. However, amino acid metabolomics analysis revealed that no amino acid metabolites remained significant after FDR correction, and thus the observed pathway-level changes (e.g., BCAA metabolism and tryptophan–kynurenine pathway) should be interpreted as nominal and exploratory findings. Overall, the results indicate that feeding systems were associated with alterations in the lipid and amino acid metabolic profiles of the biceps femoris muscle in Tibetan sheep, which were further associated with differences in muscle WHC, shear force, lipid nutritional composition, and the profile of flavor precursors. This study provides a theoretical basis for optimizing plateau meat sheep production systems and developing high-quality Tibetan sheep meat products. Full article

(This article belongs to the Section Small Ruminants)

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19 pages, 2979 KB

Open AccessArticle

β-Casein Polymorphism in Serbian Holstein-Friesian and Busha Cattle and Its Association with Milk Production Traits

by Marko Ristanic, Uros Glavinic, Jevrosima Stevanovic, Jovana Stamenovic, Nemanja M. Jovanovic, Milan Maletic and Zoran Stanimirovic

Animals 2026, 16(13), 2052; https://doi.org/10.3390/ani16132052 - 3 Jul 2026

Abstract

Interest in β-casein polymorphism and A2 milk production has increased considerably in recent years because of their potential effects on milk quality and dairy cattle breeding. The aim of this study was to determine the distribution of β-casein A1/A2 genotypes in Holstein-Friesian (HF) [...] Read more.

Interest in β-casein polymorphism and A2 milk production has increased considerably in recent years because of their potential effects on milk quality and dairy cattle breeding. The aim of this study was to determine the distribution of β-casein A1/A2 genotypes in Holstein-Friesian (HF) cattle and indigenous Busha cattle from Serbia and to evaluate the association between β-casein genotypes and milk production traits in HF cows. The study included 334 HF cows, 58 HF bulls, and 64 Busha cows. Genotyping was performed using the ACRS-PCR method, followed by restriction fragment analysis. Milk production data for HF cows included milk yield, milk protein content, and milk fat content during the first and second lactation. Statistical analysis included determination of genotype and allele frequencies, testing of Hardy–Weinberg equilibrium, and evaluation of associations between genotypes and milk production traits. In HF cows, genotype frequencies were 14.07% for A1A1, 60.18% for A1A2, and 25.75% for A2A2, while allele frequencies were 0.442 for A1 and 0.558 for A2. Similar genotype and allele distributions were observed in HF bulls, where the A2 allele frequency (0.560) was slightly higher than the A1 allele frequency (0.440). In Busha cows, a pronounced predominance of the A2 allele (0.719) and A2A2 genotype (56.25%) was detected. HF cows carrying the A2A2 genotype were associated with significantly higher milk yield, as well as higher milk protein and milk fat content, compared to cows with other genotypes (p < 0.05; p < 0.01). Milk production traits were generally higher during the second lactation, regardless of genotype. HF cows carrying the A2A2 genotype generally showed higher milk yield and more favorable milk composition traits. In addition, the high frequency of the A2 allele in Busha cattle indicates that this indigenous breed may represent a valuable genetic resource for the conservation and utilization of the A2 allele in future breeding programs. Full article

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

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