Animals

15 pages, 1267 KB

Open AccessArticle

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

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

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

Viewed by 772

Abstract

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

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

F_{S T}

,

θ_{π}

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

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

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26 pages, 6574 KB

Open AccessArticle

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

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

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

Viewed by 1304

Abstract

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

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

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

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24 pages, 3611 KB

Open AccessCase Report

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

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

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

Cited by 1 | Viewed by 2105

Abstract

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

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

(This article belongs to the Section Mammals)

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12 pages, 501 KB

Open AccessArticle

Selenium Content of the Gonads of the Domestic Dog (Canis lupus f. familiaris) in Relation to Sex, Age, and Reproductive Disorders

by Ewa Skibniewska, Marta Kołnierzak, Bartosz Skibniewski, Iwona Lasocka and Michał Skibniewski

Animals 2025, 15(23), 3502; https://doi.org/10.3390/ani15233502 - 4 Dec 2025

Viewed by 525

Abstract

Appropriate selenium (Se) levels contribute to the weakening of the effects of cellular peroxidation and have a protective function, ensuring the effectiveness of the defense against various diseases. The study aimed to determine the selenium content in the gonads of dogs from the [...] Read more.

Appropriate selenium (Se) levels contribute to the weakening of the effects of cellular peroxidation and have a protective function, ensuring the effectiveness of the defense against various diseases. The study aimed to determine the selenium content in the gonads of dogs from the Warsaw metropolitan area, taking into account the influence of gender, health status, and individual. The research material consisted of testicular and ovarian samples obtained from 86 animals during routine surgeries. Selenium content in the samples was determined using fluorometric spectrometry. The mean selenium content in the gonads of all components was found to be 0.43 mg·kg⁻¹ wet weight. In males, it was 0.49 mg·kg⁻¹ wet weight, while in females, it was 0.39 mg·kg⁻¹ wet weight. The lowest selenium concentration was found in animals of both sexes in the group of individuals older than 7 years. In individuals with disturbed selenium homeostasis, 0.34 mg·kg⁻¹ wet weight occurred in the ovaries of females with pyometra, and in males, 0.41 mg·kg⁻¹ wet weight occurred in individuals with testicular atrophy. Full article

(This article belongs to the Section Companion Animals)

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26 pages, 3317 KB

Open AccessArticle

Blood Transcriptome Analysis Reveals Immune Characteristics of Captive Forest Musk Deer (Moschus berezovskii) at High Altitude in Bianba County, Tibet

by Lei Chen, Xuxin Li, Zhoulong Chen, Jin Bai, Yanni Zhao, Maoyuan Gan, Wenjingyi Chang, Jieyao Cai and Xiuyue Zhang

Animals 2025, 15(23), 3501; https://doi.org/10.3390/ani15233501 - 4 Dec 2025

Viewed by 667

Abstract

The hypoxic, cold, and high-ultraviolet radiation environments at high altitude pose severe challenges to mammalian immune and metabolic systems. However, little is known about how captive forest musk deer adapt to high-altitude environments and their seasonal variations. This study analyzed peripheral blood transcriptomes [...] Read more.

The hypoxic, cold, and high-ultraviolet radiation environments at high altitude pose severe challenges to mammalian immune and metabolic systems. However, little is known about how captive forest musk deer adapt to high-altitude environments and their seasonal variations. This study analyzed peripheral blood transcriptomes of 33 captive forest musk deer (Moschus berezovskii) at high altitude (~3900 m) and low altitude (~1450 m) during autumn-winter and spring-summer seasons. Results revealed comprehensive immune suppression in the high-altitude group during autumn-winter (downregulation of complement system CFB/C2/C3, interferon pathway genes including FLT3, with only natural killer (NK) cell PRKCQ upregulated), coupled with energy-conserving metabolic reprogramming (altered carbohydrate metabolism, inhibited lipid synthesis, fat mobilization, suppressed protein degradation). During spring-summer, neutrophil antimicrobial responses (SLPI/NCF1/ELANE) and humoral immunity (B cell differentiation genes PAX5/RUNX1; class-switch enzyme AICDA) partially recovered while cellular immunity (IL15/B2M) remained suppressed, accompanied by enhanced anabolic metabolism and adipocyte differentiation. Notably, NK cell-mediated cytotoxicity showed selective enhancement despite comprehensive immune suppression, representing an energy-efficient innate defense strategy. This study provides the first characterization of seasonal immune dynamics in a high-altitude cervid species. These findings reveal persistent immune constraints in high-altitude populations and provide theoretical foundations for disease prevention and health management in captive forest musk deer at high altitudes. Full article

(This article belongs to the Special Issue Environmental Physiology and Health in Animals: Mechanisms, Adaptations and Welfare)

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16 pages, 7999 KB

Open AccessArticle

Patterns of Agricultural Crop Damage by Wild Boar (Sus scrofa) in South-Western Poland

by Bogusław Bobek, Anna Chrzan, Jakub Furtek, Małgorzata Kłyś, Dorota Merta and Marta Wojciuch-Płoskonka

Animals 2025, 15(23), 3500; https://doi.org/10.3390/ani15233500 - 4 Dec 2025

Viewed by 771

Abstract

Studies on agricultural crop damage inflicted by wild boar (Sus scrofa) were conducted in hunting districts located in Lower Silesia, south-western Poland. The statistical analysis of damaged agricultural crops was based on documentation obtained via survey from hunting associations managing 81 [...] Read more.

Studies on agricultural crop damage inflicted by wild boar (Sus scrofa) were conducted in hunting districts located in Lower Silesia, south-western Poland. The statistical analysis of damaged agricultural crops was based on documentation obtained via survey from hunting associations managing 81 hunting districts. For each hunting district (mean area 43.1 km²), this documentation detailed the area of damaged crops (maize, various cereals, root crops, rapeseed, grasslands, and other crops), the date of the damage, and the value of compensation paid to farmers. During three consecutive hunting seasons (2013/14–2015/16) the area of damaged farmlands was amounted to 2098.2 hectares. Maize constitutes 43.9% of the total damage area, while for various cereal crops and grasslands, the figures were 29.5% and 13.2%, respectively. The mean damage compensation per hectare amounted to €421.0, the highest being for root crops (€942.8/ha) and the lowest for grasslands (€214.8/ha). A positive correlation was shown between the wild boar harvest rate and the percentage of farmland area damaged by these animals. Between the 2015/16 and 2022/23 hunting seasons, a decline in the density of harvested wild boar from 1.99 to 1.05 individuals reduced the area of damage from 0.335 ha/km² to 0.164 ha/km² of farmland. Population density control has been suggested as the most effective method of protecting agricultural crops against wild boar. Full article

(This article belongs to the Section Wildlife)

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10 pages, 1314 KB

Open AccessFeature PaperArticle

Equine Infectious Anemia Virus in Equids: A Large-Scale Serosurvey in Western Europe

by Moisés Gonzálvez, Juan J. Franco, David Cano-Terriza, Jesús Barbero-Moyano, Eduard Jose-Cunilleras, Jesús García, Eduardo Alguacil and Ignacio García-Bocanegra

Animals 2025, 15(23), 3499; https://doi.org/10.3390/ani15233499 - 4 Dec 2025

Viewed by 679

Abstract

Background: Equine Infectious Anemia (EIA) is a notifiable disease caused by Equine Infectious Anemia Virus (EIAV), with major sanitary and economic importance for equids worldwide. There is limited and outdated information on the circulation of EIAV in many European countries. In the present [...] Read more.

Background: Equine Infectious Anemia (EIA) is a notifiable disease caused by Equine Infectious Anemia Virus (EIAV), with major sanitary and economic importance for equids worldwide. There is limited and outdated information on the circulation of EIAV in many European countries. In the present study, we aimed to assess virus exposure in different equid species in Western Europe. Methods: Between 2011 and 2023, a cross-sectional study was conducted to investigate the seroprevalence of EIAV in 1676 equids (1444 horses, 106 donkeys and 126 mules/hinnies) from four European regions: Andalusia (Southern Spain; n = 808), Catalonia (Northeastern Spain; n = 437), Southeastern United Kingdom (UK; n = 209), and Ireland (n = 222). Results: None of the analyzed equines tested positive for antibodies against EIAV (0%; 95 %CI: 0.0–0.18), indicating limited virus exposure of the equid populations from these European regions. Conclusions: This finding is consistent with the low EIAV circulation previously reported in Western Europe and the few official EIA outbreaks recorded in the region during the last decade. To the best of the author’s knowledge, this serosurvey represents the first investigation assessing EIAV exposure in equids from Ireland and the UK. Our results not only provide novel epidemiological evidence for the four European regions assessed but also reinforce the effectiveness of surveillance strategies and control measures implemented by European countries, in accordance with national and international regulations, to successfully limit EIAV circulation. Full article

(This article belongs to the Special Issue Prevention and Control for Animal Transmissible Diseases)

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17 pages, 305 KB

Open AccessArticle

Exploring Tectona grandis Linn. f. Leaf Extract as a Functional Feed Additive with Antioxidant and Nutraceutical Potential for Livestock

by Nattaya Montri, Metha Wanapat, Sungchhang Kang, Seangla Cheas, Anusorn Cherdthong, Pongsatorn Gunun, Nirawan Gunun, Suban Foiklang, Phongthorn Kongmun, Dutsadee Srithat, Pongsathorn Tongkasee and Sineenart Polyorach

Animals 2025, 15(23), 3498; https://doi.org/10.3390/ani15233498 - 4 Dec 2025

Cited by 1 | Viewed by 651

Abstract

This study investigated the phytochemical composition, antioxidant activity, and safety of Tectona grandis leaf extracts at four maturity stages. Ethanolic extracts were screened for secondary metabolites and analyzed using GC–MS, together with heavy metal determination, phenolic profiling, and antioxidant assays. Preliminary phytochemical screening [...] Read more.

This study investigated the phytochemical composition, antioxidant activity, and safety of Tectona grandis leaf extracts at four maturity stages. Ethanolic extracts were screened for secondary metabolites and analyzed using GC–MS, together with heavy metal determination, phenolic profiling, and antioxidant assays. Preliminary phytochemical screening revealed the presence of alkaloids, flavonoids, glycosides, tannins, saponins, and coumarins, with clear variation among leaf stages. GC–MS analysis detected multiple bioactive constituents, including phytol, n-decanoic acid, and heptadecenal. These compounds have been previously reported to exhibit antioxidant, antimicrobial, and anti-inflammatory properties. Heavy metal analysis showed very low cadmium concentrations (0.001–0.004 mg/kg), undetectable lead levels (<LOD), and trace levels of arsenic concentrations (0.012–0.018 mg/kg), all of which were within safe limits for plant materials. Quantitative assays demonstrated that mature leaves contained the highest total phenolic (8.751 ± 0.018 mg GAE/g DW), total flavonoid (0.359 ± 0.017 mg QE/g DW), and condensed tannin (0.303 ± 0.000 mg CE/g DW) contents. Correspondingly, mature-leaf extracts exhibited the greatest antioxidant capacity, showing 95.88 ± 0.12% inhibition of oxidant activity. The ethanolic extracts exhibited IC₅₀ values ranging from 22.93–50.35 mg/mL, whereas the ascorbic acid standard showed an IC₅₀ of 0.0767 mg/mL, confirming the expected lower potency of crude plant extracts. The strong correlation between phenolic accumulation and antioxidant activity suggests enhanced defense against oxidative stress. The absence of detectable heavy metal contamination, together with the high polyphenolic content, suggests that T. grandis leaves may serve as a safe natural source of antioxidants. These findings demonstrate the strong antioxidant potential of T. grandis leaf extracts, providing a basis for future studies evaluating their functional applications in biological systems. Full article

(This article belongs to the Special Issue Nutraceuticals and Functional Foods: Advancing Metabolic, Reproductive and Immune Health in Animals)

19 pages, 562 KB

Open AccessArticle

Clinical and Diagnostic Features of Feline Epilepsy: Distribution of Seizure Types and Associated Factors

by Martinas Jankauskas, Julija Tamosauskaite, Aistė Gradeckienė, Sigitas Čižinauskas, Dmitrij Kvitka and Vita Riškevičienė

Animals 2025, 15(23), 3497; https://doi.org/10.3390/ani15233497 - 4 Dec 2025

Viewed by 1445

Abstract

Epilepsy is a common neurological disorder in cats, but its seizure types, etiologies, and clinical associations remain incompletely characterized compared to dogs. This retrospective study analyzed 118 feline epilepsy cases from two veterinary hospitals in Finland and Lithuania, aiming to evaluate the distribution [...] Read more.

Epilepsy is a common neurological disorder in cats, but its seizure types, etiologies, and clinical associations remain incompletely characterized compared to dogs. This retrospective study analyzed 118 feline epilepsy cases from two veterinary hospitals in Finland and Lithuania, aiming to evaluate the distribution of seizure types and their associations with demographic, clinical, neurological, diagnostic, and diagnostic test results. Seizures were classified as single generalized (SG), cluster seizures (CS), or status epilepticus (SE). SG and CS occurred with nearly equal frequency (44.9% and 45.8%, respectively), while SE was less common (9.3%). Median age at presentation was 72 months, and no significant differences were found in age, sex, reproductive status, or body weight between seizure types. Abnormal clinical findings (p = 0.009) and neurological abnormalities (p < 0.001) were significantly more frequent in cats with SE, indicating greater neurological compromise in severe seizure forms. MRI was performed in 74.6% of cases and revealed a significant difference in utilization among seizure types (p = 0.004), with lower imaging rates in SE, likely reflecting clinical instability at presentation. Etiology of seizure development was established in 89 cats: idiopathic epilepsy (IE) predominated (80.9%), followed by structural (15.7%) and metabolic/toxic (3.4%) causes. Seizure type and etiology were significantly associated (p = 0.006), with structural epilepsy most common in SE and idiopathic epilepsy prevailing in SG and CS. Among cats with known outcomes (n = 35), seizure control was achieved in 68.6%, while mortality and euthanasia were higher in SE. These findings provide new insight into feline epilepsy by linking seizure type to clinical and etiological profiles, emphasizing that severe presentations are more often associated with neurological abnormalities and structural causes, whereas demographic factors alone do not predict seizure severity. Full article

(This article belongs to the Section Companion Animals)

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19 pages, 8490 KB

Open AccessFeature PaperArticle

Study of the Skull and Brain in a Cape Genet (Genetta tigrina) Using Computed Tomography and Magnetic Resonance Imaging

by Giuseppe Barillaro, Antonino Marcianò, Stella Costa, Matteo Marino, Simone Minniti, Claudia Dina Interlandi and Filippo Spadola

Animals 2025, 15(23), 3496; https://doi.org/10.3390/ani15233496 - 4 Dec 2025

Viewed by 537

Abstract

The Cape Genet (Genetta tigrina) is one of the exotic mammal species that are becoming popular as pets. Although it is possible to find some references in the literature regarding Genetta spp. and viverrids, the information about this animal remains scarce [...] Read more.

The Cape Genet (Genetta tigrina) is one of the exotic mammal species that are becoming popular as pets. Although it is possible to find some references in the literature regarding Genetta spp. and viverrids, the information about this animal remains scarce and incomplete, starting from the anatomy, a discipline that lays the foundations for deeper knowledge of a species. The aim of the present study is to identify head structures of the Cape Genet, including the skull, brain components, and the dental formula, providing for the first time a three-dimensional anatomical model of the skull, thanks to the help of advanced diagnostic imaging techniques such as computed tomography (CT) and magnetic resonance imaging (MRI), defining the ideal parameters and protocols of CT, MRI, and anesthesia for this procedure in this species. Results highlight the value of CT and MRI as study tools, enabling clear visualization of cranial structures and offering a potential reference atlas for clinicians and researchers. Full article

(This article belongs to the Special Issue Exploring Advanced Techniques in Radiography and Ultrasonography in Veterinary Medicine)

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17 pages, 2307 KB

Open AccessArticle

Proteome Analysis of Spermathecal Fluid and Seminal Plasma Reveals the Mechanism of Sperm Storage in Amphioctopus Fangsiao

by Xiaojie Sun, Jiantao Yao, Zexin Huang, Yan Li, Qihao Luo, Weijun Wang, Guohua Sun, Xiaohui Xu, Zan Li, Bin Li, Yanwei Feng and Jianmin Yang

Animals 2025, 15(23), 3495; https://doi.org/10.3390/ani15233495 - 4 Dec 2025

Viewed by 579

Abstract

The development of males and females of the cephalopod Amphioctopus fangsiao is asynchronous. The male produces sperm after maturity for storage in a spermatophore prior to mating. After mating, the sperm enter the female spermatheca for storage until ovulation occurs, a period that [...] Read more.

The development of males and females of the cephalopod Amphioctopus fangsiao is asynchronous. The male produces sperm after maturity for storage in a spermatophore prior to mating. After mating, the sperm enter the female spermatheca for storage until ovulation occurs, a period that lasts for 8 months. This is a biologically uncommon phenomenon because sperm cells generally fail to maintain their ability to fertilize for a long time after being ejaculated. However, the molecular mechanisms of this phenomenon are still not clear. Sperm cells are stored in the male spermatophore and the female spermatheca, each of which provides a suitable environment. To determine the molecular basis of the sperm storage mechanisms in A. fangsiao, protein profiles from spermathecal fluid and seminal plasma were characterized separately using mass spectrometry-based proteomics. The antioxidant enzymes superoxide dismutase (SOD), glutathione S-transferase (GST), and Thioredoxin (Trx), and the glycolytic enzymes lactate dehydrogenase (LDH), hexokinase (HK), pyruvate dehydrogenase kinase (PDK), and ATP synthase were significantly enriched in the spermathecal fluid. Catalase (CAT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), triosephosphate isomerase (TIM), phosphoglycerate kinase (PGK), and Chitinase were significantly enriched in the seminal plasma. The antimicrobial proteins transforming growth factor beta regulator 1 (TBRG1) and interleukin enhancer binding factor 2 (ILF2) and the extracellular matrix-related proteins transforming growth factor beta induced protein (TGFBIp) and thrombospondin type-1 domain-containing protein 4 (THSD4) were also significantly expressed in the spermathecal fluid. These proteins may be crucial for successful long-term sperm storage. We measured the activities of four antioxidant enzymes based on the proteomic results, supporting the antioxidant mechanism during the sperm storage process. This study enhances our understanding of the sperm storage ability of A. fangsiao. Full article

(This article belongs to the Section Aquatic Animals)

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11 pages, 1614 KB

Open AccessArticle

The Role of Dietary Schizochytrium Powder in Chicken Production Performance, Egg Quality, and Antioxidant Status

by Qianbao Wang, Huayun Huang, Chunmiao Li, Zhengyang Huang, Zhaolin Wu, Linglin Kong, Zhenhua Zhao and Zhicheng Wang

Animals 2025, 15(23), 3494; https://doi.org/10.3390/ani15233494 - 4 Dec 2025

Viewed by 513

Abstract

Objectives: The aim of this experiment was to explore how the addition of Schizochytrium powder to the feeding ration affected the production performance, egg quality, and antioxidant function of chickens. Schizochytrium powder is a unicellular spherical marine microalga that can be cultivated through [...] Read more.

Objectives: The aim of this experiment was to explore how the addition of Schizochytrium powder to the feeding ration affected the production performance, egg quality, and antioxidant function of chickens. Schizochytrium powder is a unicellular spherical marine microalga that can be cultivated through heterotrophic fermentation, with characteristics including rapid cell growth, stable composition, and ease of large-scale production. Experimental design: Three hundred and twenty 33-week-old chickens with similar egg production rates and body weights were selected and randomly divided into four groups, with five replicates each and 16 hens in each replicate. The control group (Group I) was fed a corn–soybean meal basal diet, while the test groups were supplemented with 0.5% (Group II), 1.0% (Group III), and 2.0% (Group IV) Schizochytrium powder on top of this basal diet, respectively. The pretest period was 1 week, and the main test period was 8 weeks. The results revealed the following: (1) Compared with Group I, the average daily feed intake (ADFI) and laying rate (LR) were significantly lower (p < 0.05) in Group IV, and there was no significant difference (p > 0.05) in the ADFI and LR between Groups II and III. (2) Compared with Group I, the eggshell strength (ES) and docosahexaenoic acid (DHA) content of Groups II, III, and IV were significantly higher (p < 0.05). (3) Compared with Group I, serum triglyceride (TG) content was significantly decreased (p < 0.05), and serum low-density lipoprotein (LDL) content was significantly increased (p < 0.05) in Groups II, III, and IV. Serum total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and peroxidase (PO) activities were significantly higher (p < 0.05) and serum malondialdehyde (MDA) content was significantly lower (p < 0.05) in Groups II, III, and IV compared to Group I. In conclusion, adding Schizochytrium powder to the feeding ration could affect chickens’ production performance, increase egg DHA content, and improve the antioxidant capacity of the organism. Based on the results of this study, we recommend a ratio of 1.0% Schizochytrium powder addition. Full article

(This article belongs to the Special Issue A Paradigm Shift—Advances in Poultry Nutrition and Management, and Alternative Poultry Feed Ingredients Using Novel Feed Processing Methods: Second Edition)

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14 pages, 1551 KB

Open AccessArticle

Acoustic Diversity in Zhangixalus lishuiensis: Intra-Individual Variation, Acoustic Divergence, and Genus-Level Comparisons

by Jia-Jun Hao, Zhi-Qiang Chen, Hua-Li Hu, Jian-Guo Cui and Guo-Hua Ding

Animals 2025, 15(23), 3493; https://doi.org/10.3390/ani15233493 - 4 Dec 2025

Viewed by 444

Abstract

Advertisement calls play a crucial role in anuran species recognition and mate selection, yet acoustic characteristics of Zhangixalus lishuiensis remain undescribed. This study presents the first detailed acoustic analysis of advertisement calls in Zhangixalus lishuiensis from Lishui, Zhejiang Province, China. Based on recordings [...] Read more.

Advertisement calls play a crucial role in anuran species recognition and mate selection, yet acoustic characteristics of Zhangixalus lishuiensis remain undescribed. This study presents the first detailed acoustic analysis of advertisement calls in Zhangixalus lishuiensis from Lishui, Zhejiang Province, China. Based on recordings from 13 males collected in the field during the breeding season, we documented a hierarchical call structure consisting of three note types (Initial Note, Middle Note, and Short Note) organized into two note groups (Note Group 1 (NG1) and Note Group 2). Four structural variants of NG1 were identified, revealing intra-individual acoustic diversity within the population. Compared to the closely related Z. zhoukaiyae from the Dabie Mountains population, male Z. lishuiensis produced calls with significantly longer temporal parameters but lower dominant frequency, despite overall structural similarity, indicating acoustic divergence between these taxa. Comparative analysis across eleven Zhangixalus species revealed substantial variation in call durationand dominant frequency. These findings enrich the bioacoustic characterization of Zhangixalus and establish a foundation for acoustic-based species identification, monitoring, and taxonomic studies in this genus. Full article

(This article belongs to the Section Herpetology)

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21 pages, 2563 KB

Open AccessArticle

Molecular Epidemiology and Antibiotic Resistance of Sheep-Derived Mannheimia haemolytica in Northwestern China

by Chenxiao Wang, Leina Dou, Juan Wang, Dongyang Ye and Zengqi Yang

Animals 2025, 15(23), 3492; https://doi.org/10.3390/ani15233492 - 4 Dec 2025

Viewed by 681

Abstract

This study comprehensively explored the epidemiology, virulence, antibiotic resistance, and genomic features of ovine-derived Mannheimia haemolytica across three provinces in Northwest China (Gansu, Ningxia, and Shaanxi). Lung tissue samples were collected postmortem from sheep with respiratory disease (nine per province), cultured under sterile [...] Read more.

This study comprehensively explored the epidemiology, virulence, antibiotic resistance, and genomic features of ovine-derived Mannheimia haemolytica across three provinces in Northwest China (Gansu, Ningxia, and Shaanxi). Lung tissue samples were collected postmortem from sheep with respiratory disease (nine per province), cultured under sterile conditions, and screened by morphological, Gram staining, PCR, and NGS methods. Nine M. haemolytica strains were successfully isolated (isolation rate 33.33%) and identified as serotypes A1, A2, and A6, with Shaanxi exhibiting a significantly higher isolation rate of 66.67%. Genomic analysis revealed that each isolate each harbored over 100 virulence genes and displayed notable genetic diversity. In murine assays, intraperitoneal inoculation in mice caused severe breathing problems and death within 24 h; necropsy revealed lung congestion, splenic necrosis foci, and hepatic congestion. Antibiotic susceptibility testing showed sensitivity to ciprofloxacin, azithromycin, gentamicin, and levofloxacin, while resistance was observed for tiamulin and enrofloxacin. These findings indicate a high prevalence and strong pathogenicity of ovine M. haemolytica in Shaanxi, underscore the presence of key resistance traits, and provide a genomic and pathogenic reference to guide region-specific surveillance, therapeutic decisions, and vaccine-oriented control in small ruminant herds. Importantly, these results highlight the need for region-specific monitoring and judicious antibiotic use, which are essential to forestall the spread of resistant strains and to ensure sustainable disease management strategies. Full article

(This article belongs to the Section Veterinary Clinical Studies)

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18 pages, 7600 KB

Open AccessArticle

Quercetin Enhances the Antibacterial Activity of Polymyxin E Against MCR-1-Positive Bacteria by Inhibiting the Biological Functions of the Cell Membrane

by Yongjie Zhang, Liyang Guo, Shun Wang, Jie Zhang, Xinlei Ren, Rui Li, Jichang Li and Chunli Chen

Animals 2025, 15(23), 3491; https://doi.org/10.3390/ani15233491 - 3 Dec 2025

Viewed by 656

Abstract

In animal husbandry and livestock farming, with the spread of the plasmid-mediated MCR-1 gene, polymyxin E, as the last line of defense against drug-resistant Gram-negative bacteria, is facing severe challenges. This study investigated the in vitro and in vivo synergistic effects and mechanisms [...] Read more.

In animal husbandry and livestock farming, with the spread of the plasmid-mediated MCR-1 gene, polymyxin E, as the last line of defense against drug-resistant Gram-negative bacteria, is facing severe challenges. This study investigated the in vitro and in vivo synergistic effects and mechanisms of QUE combined with polymyxin E against MCR-1-positive chicken E. coli JD37. In vitro experiments showed that QUE could restore the sensitivity of E. coli JD37 to polymyxin E (FIC = 0.34375) and enhance the bactericidal effect of polymyxin E by increasing cell membrane permeability, fluidity, and membrane potential, downregulating the expression of the AcrAB-TolC efflux pump and LPS-related genes. Molecular docking further identified the key residues for QUE binding to the MCR-1 protein. The in vivo chick infection model confirmed that combination therapy increased survival rates, reduced bacterial load in tissues, alleviated pathological damage, and decreased levels of intestinal inflammatory factors. Our results demonstrate the synergistic bactericidal effect of the QUE-polymyxin E combination against MCR-1-positive E. coli and elucidate its underlying mechanism. Full article

(This article belongs to the Section Poultry)

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17 pages, 2190 KB

Open AccessArticle

Spatiotemporal Niche Differentiation of Ungulates in the Southwest Mountains, China

by Qingsong Jiang, Hangshu Xiao, Huaqiang Zhou, Ying Li, Jinghui Fu, Assan Meshach, Qiuxian Li, Liwen Kang, Li Yan, Yixin Shu, Jing Zhang, Zejun Zhang, Mingsheng Hong and Jianmei Xie

Animals 2025, 15(23), 3490; https://doi.org/10.3390/ani15233490 - 3 Dec 2025

Viewed by 648

Abstract

Spatiotemporal niche differentiation plays a critical role in facilitating mutual adaptation and sustaining coexistence among sympatric species. We investigated these patterns in sympatric ungulates through an infrared camera trap survey conducted in the Kazila Mountain region of southwestern China from July 2023 to [...] Read more.

Spatiotemporal niche differentiation plays a critical role in facilitating mutual adaptation and sustaining coexistence among sympatric species. We investigated these patterns in sympatric ungulates through an infrared camera trap survey conducted in the Kazila Mountain region of southwestern China from July 2023 to May 2025. A total of seven species were recorded across 54 camera sites, with tufted deer (Elaphodus cephalophus) being the most frequently detected, while forest musk deer (Moschus berezovskii) and Chinese goral (Naemorhedus griseus) were the least. Nocturnality indices (β > 0.54 indicating nocturnal, β < 0.54 indicating diurnal, and β = 0.54 indicating no distinct diel preference) revealed significant differences in activity patterns among the five species. Tufted deer (β = 0.415), alpine musk deer (Moschus chrysogaster) (β = 0.438), and wild boar (Sus scrofa) (β = 0.234) were predominantly diurnal. In contrast, sambar (Rusa unicolor) (β = 0.571) was nocturnal, while the Chinese serow (Capricornis milneedwardsii) (β = 0.534) showed no strong diel preference. Nine of ten species pairs exhibited significant diel rhythm differences, with the exception of sambar-Chinese serow, and these rhythms showed marked seasonal variation, particularly in tufted deer, Chinese serow, and sambar. Temporal overlap was generally higher in the cold season for seven species pairs, suggesting that such overlap may be related to resource availability and increased interspecific competition under harsher conditions. Pianka’s overlap index (Oik) (ranging from 0 to 1, where 0 indicates no overlap and 1 indicates complete overlap) was used to assess spatial niche overlap, with values ranging from 0.16 (alpine musk deer–wild boar) to 0.86 (tufted deer–wild boar). Spatial autocorrelation and clustering analysis showed that tufted deer exhibited significant positive spatial autocorrelation, indicating a clustered high-value distribution, while the other species were randomly distributed. Spatial hotspot analysis revealed substantial overlap between tufted deer and wild boar, while the remaining species showed higher levels of spatial segregation. Collectively, these results suggest that seasonal variation in activity patterns, coupled with spatial segregation, mitigates interspecific competition and supports the stable sympatric coexistence of ungulates in this montane ecosystem. Full article

(This article belongs to the Section Wildlife)

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15 pages, 1607 KB

Open AccessArticle

The Usefulness of Extemporaneous Cytological Examination in Imaging-Guided Biopsies

by Andrea Rubini, Francesca Del Signore, Massimo Vignoli, Arianna Miglio, Martina Rosto, Andrea De Bonis, Rossella Terragni, Domenico Santori and Morena Di Tommaso

Animals 2025, 15(23), 3489; https://doi.org/10.3390/ani15233489 - 3 Dec 2025

Viewed by 459

Abstract

This study investigates the usefulness of extemporaneous cytological examination performed during imaging-guided biopsies in dogs and cats. In veterinary diagnostics, imaging techniques such as ultrasound and computed tomography are essential for identifying lesions, but they cannot distinguish between inflammatory and neoplastic processes. Biopsies [...] Read more.

This study investigates the usefulness of extemporaneous cytological examination performed during imaging-guided biopsies in dogs and cats. In veterinary diagnostics, imaging techniques such as ultrasound and computed tomography are essential for identifying lesions, but they cannot distinguish between inflammatory and neoplastic processes. Biopsies are therefore required, and this study explores whether immediate cytological evaluation of tissue cores, rolled onto slides and examined under a microscope, can help determine sample adequacy and reduce the number of biopsies needed. Seventy-nine animals with soft tissue or bone masses underwent ultrasound or CT-guided biopsies, followed by extemporaneous cytology and histopathological analysis. Cellular material representative of the lesion was identified in 81.1% of cases, allowing clinicians to conclude the procedure without further sampling. The diagnostic accuracy of cytology compared to histology was 68.3%, with particularly high accuracy for lipomas, melanomas, and mast cell tumors. When excluding non-diagnostic samples due to poor preparation or lesion characteristics, the accuracy reaches 92.1%. These findings suggest that extemporaneous cytology is a useful tool for verifying sample adequacy in real time, potentially reducing procedural risks and improving diagnostic efficiency. While histology remains the gold standard, this method offers immediate feedback and may enhance clinical decision-making in veterinary interventional diagnostics. Full article

(This article belongs to the Section Companion Animals)

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15 pages, 2218 KB

Open AccessArticle

The Conserved Roles of miR-2c in the Ecdysone Signaling Pathway by Targeting EcR/RXR and Runt for Exoskeleton Formation in the Pearl Oyster Pinctada fucata martensii

by Zhe Zheng, Weilin Gao, Yalin Xu, Hongmei Yang, Meichen Lu, Minxin Liang, Chuangye Yang and Jiawei Zhang

Animals 2025, 15(23), 3488; https://doi.org/10.3390/ani15233488 - 3 Dec 2025

Viewed by 405

Abstract

As organisms evolved from ancestral protists to multicellular animals, body size increased dramatically, necessitating the co-evolution of systems for movement and support [...] Full article

(This article belongs to the Section Aquatic Animals)

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19 pages, 1319 KB

Open AccessArticle

Effects of Corn Steep Liquor on the Fermentation Quality, Bacterial Community and Ruminal Degradation Rate of Corncob Silage

by Xinyi Wang, Xinfeng Wang, Tengyu Wang, Xiaoping Chen, Zuoxing Huang, Rui Yang, Shuai Liu, Xinwen Sun and Dengke Hua

Animals 2025, 15(23), 3487; https://doi.org/10.3390/ani15233487 - 3 Dec 2025

Viewed by 702

Abstract

This study aims to investigate the effect of varying addition levels of corn steep liquor (CSL) on the fermentation quality, bacterial community, and ruminal degradation rate of corncob silage. The experiment included a control group (CON) and four treatment groups: L1 with 5% [...] Read more.

This study aims to investigate the effect of varying addition levels of corn steep liquor (CSL) on the fermentation quality, bacterial community, and ruminal degradation rate of corncob silage. The experiment included a control group (CON) and four treatment groups: L1 with 5% CSL (50 g·kg⁻¹ fresh matter), L2 with 10% CSL (100 g·kg⁻¹ fresh matter), L3 with 15% CSL (150 g·kg⁻¹ fresh matter), and L4 with 20% CSL (200 g·kg⁻¹ fresh matter). The water content was controlled at 65% during fermentation for a period of 45 days. The results showed that the addition of CSL significantly increased the contents of dry matter (DM), crude protein (CP), and lactic acid (LA), while decreasing the pH, neutral detergent fiber (NDF), acid detergent fiber (ADF), and ammonia nitrogen (NH₃-N). Furthermore, the addition of CSL altered the relative abundance of microbial genera. While Pediococcus was the dominant bacterium in the CON group, Lactobacillus became the prevalent species upon the addition of CSL, and its relative abundance increased in accordance with the supplemental amount. These findings suggest that CSL provides a favorable environment for lactic acid bacteria. It is worth noting that CSL addition did not significantly alter the phylum-level bacterial community structure. The dominant bacterial taxa across all treatments were Bacillota, Proteobacteria, and Bacteroidota, with their cumulative relative abundance accounting for over 95%. The rumen degradation of the tested feedstuff was determined using the in situ nylon bag method. Results revealed that incorporating CSL into corncob silage significantly enhanced the effective degradation rates of DM, CP, NDF, and ADF in the rumen of Kazakh sheep. Specifically, the effective degradation rate of DM in the CON group was only 49.10%, which increased to 53.12% following the addition of 20% CSL, along with corresponding improvements in the degradation rates of CP, NDF, and ADF. In summary, as a valuable feed additive, corn steep liquor supports the proliferation of beneficial microorganisms in fermentation systems by supplying essential growth substrates. Additionally, it improves the nutritional balance of corncob feed and further enhances the absorption and utilization of nutrients from this feed by animals. Full article

(This article belongs to the Special Issue Alternative Protein Sources for Animal Feeds)

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10 pages, 9791 KB

Open AccessCase Report

Pericardial Sarcoma of Uncertain Origin in a Young Cat: A Case Report Highlighting Diagnostic Challenges and Thoracic Drainage in Palliative Management

by Miki Hirose, Kazumi Shimada, Aki Takeuchi, Kazuyuki Terai, Aimi Yokoi, Ahmed Farag, Akari Hatanaka, Rio Hayashi, Marino Hosoki, Daigo Azakami, Rina Nabeta, Ikki Mitsui, Lina Hamabe and Ryou Tanaka

Animals 2025, 15(23), 3486; https://doi.org/10.3390/ani15233486 - 3 Dec 2025

Viewed by 499

Abstract

Undifferentiated sarcoma is a malignant tumor characterized by the absence of a distinct cellular origin, as determined by morphological and immunohistochemical analyses, and is known for its highly invasive and proliferative behavior [...] Full article

(This article belongs to the Special Issue The Heart Under the Microscope: Congenital and Acquired Heart Defects in Dogs and Cats—from Diagnosis to Therapy and Beyond—Unveiling Future Frontiers)

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15 pages, 1068 KB

Open AccessArticle

Comparative Effects of Aminophylline, Caffeine, and Doxapram in Hypoxic Neonatal Dogs Born by Cesarean Section

by Júlia Cosenza Mendonça, Keylla Helena Nobre Pacífico Pereira, Gleice Mendes Xavier, Kárita da Mata Fuchs, Thaís Gomes Faustino, Viviane Maria Codognoto, Miriam Harumi Tsunemi, Regina Kiomi Takahira, Maricy Apparício and Maria Lucia Gomes Lourenço

Animals 2025, 15(23), 3485; https://doi.org/10.3390/ani15233485 - 3 Dec 2025

Viewed by 889

Abstract

Background: Perinatal hypoxia is a leading cause of early mortality in canine neonates and accounts for over 90% of early deaths in canine neonates. Respiratory depression may result from dystocia, prolonged labor, or anesthetic protocols used during cesarean delivery, requiring rapid pharmacological support. [...] Read more.

Background: Perinatal hypoxia is a leading cause of early mortality in canine neonates and accounts for over 90% of early deaths in canine neonates. Respiratory depression may result from dystocia, prolonged labor, or anesthetic protocols used during cesarean delivery, requiring rapid pharmacological support. This study evaluated the effects of aminophylline, caffeine, and doxapram on clinical and biochemical viability parameters in hypoxic neonatal dogs. Methods: Forty-five puppies born via elective or emergency cesarean section were included, of which 35 exhibiting cardiorespiratory depression (heart rate ≤ 180 bpm, respiratory rate < 15 movements per minute, Apgar score < 7, and reflex score < 3) were randomly allocated into aminophylline group (AG; n = 12), caffeine group (CafG; n = 11), or doxapram group (DG; n = 12). Ten clinically healthy neonates constituted the physiological control group (CG). Apgar score, neonatal reflexes, lactate, blood glucose, oxygen saturation, blood gas parameters, and cardiac troponin I (cTnI) were evaluated immediately after birth (M1) and 10 min following treatment (M10). Results: No significant differences were observed in laboratory parameters among groups (p > 0.05). Although all evaluated drugs improved clinical status by increasing heart rate, respiratory rate, and reflex scores, caffeine produced the greatest enhancement in vitality. The Δ Apgar score showed a significant difference between CafG and DG (p = 0.0095), while no statistical differences were detected between AG and CafG (p = 0.08) or between AG and GD (p = 0.60). Conclusions: Aminophylline, caffeine, and doxapram supported postnatal resuscitation in hypoxic neonatal puppies; however, caffeine demonstrated superior improvement in Apgar recovery, suggesting its potential as a preferred pharmacological adjunct in neonatal canine resuscitation protocols. Full article

(This article belongs to the Special Issue Canine and Feline Reproduction)

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19 pages, 2783 KB

Open AccessArticle

Grassland Saline-Alkaline Degradation-Induced Excessive Iron and Sodium Intake Potentially Increases the Transmission Risk of Fecal Pathogenic Bacteria in Cattle

by Yizhen Wang, Bingnan Gao, Guangming Ma, Man Xu, Yu Zhou and Xin Jiang

Animals 2025, 15(23), 3484; https://doi.org/10.3390/ani15233484 - 3 Dec 2025

Viewed by 382

Abstract

The environmental transmission of pathogens from domesticated ruminants presents a significant risk to the well-being of both humans and animals. However, the risk of pathogen release from grazing ruminants in natural grasslands has received comparatively little attention. The threat of pathogen release from [...] Read more.

The environmental transmission of pathogens from domesticated ruminants presents a significant risk to the well-being of both humans and animals. However, the risk of pathogen release from grazing ruminants in natural grasslands has received comparatively little attention. The threat of pathogen release from grazing ruminants may be increasing owing to grasslands in arid and semi-arid regions experiencing prevalent saline-alkaline degradation. Therefore, further risk assessment of pathogen release driven by saline-alkaline degradation is warranted. Here, we investigated the impact of saline-alkaline degradation in grasslands on the fecal pathogenic bacteria of domestic ruminant cattle by comparing cattle subjected to long-term grazing on undegraded (UG) and severe saline-alkaline degraded (SG) grasslands. Our results showed that the relative abundance of potential pathogenic bacteria including Streptococcus, Mogibacterium, and Alistipes in the SG cattle group was higher than that of the UG group. Notably, the results for correlation analyses showed that the Paludibacter had a positive effect on these three potential pathogenic bacteria. Moreover, we found that excessive intake of sodium and iron is associated with these microbes that have potential pathogenic risks. Our results indicate that the continued global saline-alkaline degradation of grasslands will significantly elevate the risk of animal diseases, and may also pose a serious threat to human health through environmentally mediated transmission. Full article

(This article belongs to the Section Cattle)

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11 pages, 984 KB

Open AccessFeature PaperArticle

A Portable Fluorometer Detects Significantly Elevated Cell-Free DNA in Tracheal Wash and Bronchoalveolar Lavage Fluid in Horses with Severe Asthma

by Bethanie L. Cooper, Kallie J. Hobbs, Rosemary Bayless, Austen Stinson-Miller, Erika Gruber, Kate Hepworth-Warren, Jean-Pierre Lavoie and M. Katie Sheats

Animals 2025, 15(23), 3483; https://doi.org/10.3390/ani15233483 - 3 Dec 2025

Viewed by 486

Abstract

Severe equine asthma (sEA) is characterized by increased lower airway neutrophils that contribute to dysregulated inflammation through the release of cytokines, reactive oxygen species and neutrophil extracellular traps (NETs). NETs are composed of cell-free DNA (cfDNA) intercalated with enzymatic proteins and are known [...] Read more.

Severe equine asthma (sEA) is characterized by increased lower airway neutrophils that contribute to dysregulated inflammation through the release of cytokines, reactive oxygen species and neutrophil extracellular traps (NETs). NETs are composed of cell-free DNA (cfDNA) intercalated with enzymatic proteins and are known to be increased in the lower airway of asthmatic horses. The objectives of this study were two-fold: 1. Determine if cfDNA can be accurately measured in equine bronchoalveolar lavage fluid (BALF) and tracheal wash (TW) with a Qubit 4 fluorometer. 2. Determine whether Qubit-measured cfDNA in BALF or TW is significantly different in horses with sEA, mild/moderate neutrophilic equine asthma, mastocytic equine asthma and healthy horses. A total of sixty-three horses received a physical examination and clinical score followed by a BAL +/− TW. Cell-free DNA was measured using three methods in unfiltered BAL and TW as well as BAL and TW supernatant. Cell-free DNA concentrations were highly correlated between the Qubit 4 fluorometer and NanoDrop spectrophotometer as well as between the Qubit 4 fluorometer and SYTOX green plate-based assay. Cell-free DNA concentrations were highly correlated between unfiltered TW and TW supernatant as well as between unfiltered BALF and BAL supernatant. Cell-free DNA concentrations in BAL and TW supernatant were significantly higher in horses with sEA compared to healthy horses or horses with mild/moderate equine asthma. Cell-free DNA is a biomarker of sEA that can be easily measured in the field with the small portable Qubit 4 fluorometer in BAL and TW fluid. These findings support further investigation of NETs as a biomarker and potential therapeutic target for severe equine asthma. Full article

(This article belongs to the Section Equids)

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12 pages, 922 KB

Open AccessReview

The Role of Short-Chain Fatty Acids (SCFAs) in Colic and Anti-Inflammatory Pathways in Horses

by Nathan Schank, Ashley Cottone, Michelle Wulf, Keely Seiter, Brinley Thomas, Lynda M. J. Miller, Stacy L. Anderson, Amal Sahyoun, Ammaar H. Abidi, Modar Kassan and Ashutosh Verma

Animals 2025, 15(23), 3482; https://doi.org/10.3390/ani15233482 - 3 Dec 2025

Cited by 1 | Viewed by 970

Abstract

Equine colic remains a prevalent and potentially life-threatening condition with multifactorial origins, including dietary imbalances, stress, and microbial dysbiosis. Central to equine gut health is the production of short-chain fatty acids (SCFAs), primarily acetate, propionate, and butyrate, generated through microbial fermentation of dietary [...] Read more.

Equine colic remains a prevalent and potentially life-threatening condition with multifactorial origins, including dietary imbalances, stress, and microbial dysbiosis. Central to equine gut health is the production of short-chain fatty acids (SCFAs), primarily acetate, propionate, and butyrate, generated through microbial fermentation of dietary fibers in the hindgut. These metabolites not only serve as vital energy sources but also play crucial roles in maintaining intestinal barrier integrity, modulating motility, and suppressing inflammation. This review explores the role of SCFAs in equine gastrointestinal health, with particular emphasis on their anti-inflammatory effects and potential to prevent or mitigate colic. We examine how SCFAs interact with immune pathways, via G-protein-coupled receptors and regulatory T-cell promotion, to reduce pro-inflammatory cytokines such as TNF-α and IL-6. Evidence suggests that dietary shifts toward high-starch or low-fiber intake can reduce SCFA production, contributing to microbial imbalance, increased gut permeability, and systemic inflammation, all hallmarks of colic pathophysiology. Strategies to enhance SCFA levels, including high-forage diets, targeted prebiotic and probiotic supplementation, and emerging approaches like fecal microbiota transplantation, are discussed. Despite promising findings, significant gaps remain in equine-specific research, highlighting the need for longitudinal and mechanistic studies. Understanding and harnessing the therapeutic potential of SCFAs could pave the way for novel, microbiome-based interventions in colic prevention and treatment. Full article

(This article belongs to the Section Equids)

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12 pages, 1423 KB

Open AccessArticle

Mathematical Modeling of In Vitro Rumen Fermentation Kinetics in Capiaçu Elephant Grass Silages with Inclusion of Dehydrated Cashew Pseudo-Fruit

by Isadora Osório Maciel Aguiar Freitas, Antonio Leandro Chaves Gurgel, Luís Carlos Vinhas Ítavo, Luiz Antônio Rodrigues, Vitor Cardoso Queiroz, Edy Vitoria Fonseca Martins, Marcos Jácome de Araújo, Tairon Pannunzio Dias-Silva, João Virgínio Emerenciano Neto and Alfonso Juventino Chay-Canul

Animals 2025, 15(23), 3481; https://doi.org/10.3390/ani15233481 - 3 Dec 2025

Viewed by 508

Abstract

This study aimed to evaluate and compare the performance of five mathematical models: Gompertz, Ørskov & McDonald, Brody, Richards, and the Dual Pool Logistic model, in describing the in vitro gas production kinetics of Capiaçu elephant grass (Pennisetum purpureum Schum ‘BRS Capiaçu’) silages. [...] Read more.

This study aimed to evaluate and compare the performance of five mathematical models: Gompertz, Ørskov & McDonald, Brody, Richards, and the Dual Pool Logistic model, in describing the in vitro gas production kinetics of Capiaçu elephant grass (Pennisetum purpureum Schum ‘BRS Capiaçu’) silages. The effect of including dehydrated cashew pseudo-fruit on the in vitro degradation curves was also assessed. A completely randomized design was adopted, using Capiaçu silages containing 0%, 10%, 20%, or 30% dehydrated cashew pseudo-fruit. Rumen fermentation kinetics were measured through cumulative in vitro gas production. Model performance was evaluated using the Akaike Information Criterion (AIC), coefficient of determination (R²), concordance correlation coefficient (CCC), and mean square prediction error (MSPE). Accuracy (pMSPE) and precision (AIC) were also considered. The Richards model performed best with the lowest AIC (1119.07) and MSPE (0.246) and the highest R² (0.917) and CCC (0.966). It was over 350 times more likely to provide a correct fit (p < 0.05) compared to the other models. Significant differences (p < 0.05) were observed between degradation curves as a function of the pseudo-fruit inclusion level. Increasing pseudo-fruit inclusion improved silage composition, raising total digestible nutrients (from 54.6% to 67.1%) and reducing neutral detergent fiber (from 58.5% to 42.3%), which directly enhanced fermentation kinetics. These results indicate that the Richards model is the most suitable for describing the fermentation kinetics of Capiaçu elephant grass silages. Moreover, linking model performance to practice, the Richards model provides a reliable tool for determining optimal inclusion levels of dehydrated cashew pseudo-fruit (up to 30%), supporting better silage nutritional quality and more efficient feed utilization in ruminant production systems. Full article

(This article belongs to the Section Animal Nutrition)

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23 pages, 1975 KB

Open AccessReview

Molecular Regulatory Mechanisms of Mammary Gland Development: A Review

by Xiangnan Zhou, Abd Ullah, Limeng Shi, Manna Dou, Changfa Wang, Muhammad Zahoor Khan, Chunming Wang and Xinhao Zhang

Animals 2025, 15(23), 3480; https://doi.org/10.3390/ani15233480 - 2 Dec 2025

Viewed by 1891

Abstract

Mammary gland development represents a complex and highly regulated biological process that is critical for mammalian reproductive success. This review provides a comprehensive analysis of the molecular, cellular, and hormonal mechanisms governing mammary gland development across distinct developmental stages: embryonic, pubertal, pregnancy, lactation, [...] Read more.

Mammary gland development represents a complex and highly regulated biological process that is critical for mammalian reproductive success. This review provides a comprehensive analysis of the molecular, cellular, and hormonal mechanisms governing mammary gland development across distinct developmental stages: embryonic, pubertal, pregnancy, lactation, and involution. Mammary organogenesis is orchestrated by multiple factors, including conserved genetic pathways, endocrine signaling cascades, and environmental stimuli. Drawing primarily from studies in rodents and dairy ruminants, recent advances in molecular biology have elucidated key signaling pathways, including Wnt, Notch, and TGF-β, which provide fundamental insights into mammary epithelial growth and differentiation. Furthermore, we examine the effects of external factors, including thermal stress, photoperiodic variation, and dry period duration on mammary function and lactational performance. The dry period, in particular, serves a critical role in tissue remodeling and preparation for subsequent lactation cycles. Through systematic review of physiological and molecular dynamics across developmental stages, this article identifies key genetic regulators and cellular mechanisms, offering valuable insights for optimizing milk production and composition in agricultural systems. Full article

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

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11 pages, 702 KB

Open AccessFeature PaperArticle

Molecular Screening of Sarcocystis spp. in Grazing Sheep (Ovis aries) and Shepherd Dogs (Canis lupus familiaris) from Central Portugal

by Sara Gomes-Gonçalves, Ricardo J. Figueiredo, Soraia Rodrigues, Jaqueline T. Bento, Sérgio Santos-Silva, Daniela Almeida, Rita Cruz, Fernando Esteves, Alexandra Lameira Baptista, Maria Aires Pereira, Luís Cardoso and João R. Mesquita

Animals 2025, 15(23), 3479; https://doi.org/10.3390/ani15233479 - 2 Dec 2025

Viewed by 592

Abstract

Sarcocystis spp. are cyst-forming protozoan parasites with a global distribution that infect a wide range of domestic and wild animals, impacting both animal health and livestock productivity. In sheep, infections can cause clinical disease, reproductive losses, and economic damage, particularly when pathogenic species [...] Read more.

Sarcocystis spp. are cyst-forming protozoan parasites with a global distribution that infect a wide range of domestic and wild animals, impacting both animal health and livestock productivity. In sheep, infections can cause clinical disease, reproductive losses, and economic damage, particularly when pathogenic species such as Sarcocystis tenella are involved. Grazing sheep, including breeds such as the Serra da Estrela from central Portugal, are at increased risk due to frequent contact with shepherd dogs, which serve as definitive hosts. Despite their significance, data on the occurrence and distribution of Sarcocystis spp. in Portuguese sheep remain limited. This study analyzed 179 samples collected in central Portugal during 2024, including 41 brain tissues and 88 blood samples from sheep, and 50 stool samples from shepherd dogs, using conventional PCR and bidirectional Sanger sequencing. Sarcocystis sp. closely related to S. tenella was detected exclusively in sheep brain tissue, with a prevalence of 4.9% (2/41; 95% CI: 0.60–16.53), while no parasite DNA was found in blood or dog samples. These results provide the first molecular confirmation of Sarcocystis spp. closely related to S. tenella in Portuguese sheep raised for human consumption and establish baseline data for future epidemiological surveillance and control strategies. Full article

(This article belongs to the Special Issue Infectious Diseases in Animals: Community Ecology and Pathogen Dynamics, 2nd Edition)

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19 pages, 5360 KB

Open AccessArticle

Characterizing the Norway Lobster Nephrops norvegicus (L., Homarida: Decapoda) Maturity of Males in the Mediterranean Sea: Morphological and Physiological Aspects

by Cristina Porcu, Noemi Pascale, Andrea Bellodi, Pierluigi Carbonara, Alessandro Cau, Riccardo Demurtas, Antonello Mulas and Maria Cristina Follesa

Animals 2025, 15(23), 3478; https://doi.org/10.3390/ani15233478 - 2 Dec 2025

Viewed by 575

Abstract

In male decapods, such as the Norway lobster Nephrops norvegicus, sexual maturity is not easily determined by macroscopic examination of gonads and few studies have been conducted on this topic. Sexual maturity is linked to changes in both external morphology and physiology. [...] Read more.

In male decapods, such as the Norway lobster Nephrops norvegicus, sexual maturity is not easily determined by macroscopic examination of gonads and few studies have been conducted on this topic. Sexual maturity is linked to changes in both external morphology and physiology. This study analyzed the maturity of N. norvegicus males in the Central-Western Mediterranean Sea (Sardinia), focusing on primary sexual characteristics (testes and vasa deferentia) and morphometric traits (appendix masculina and petasma). Since the testes showed no macroscopic changes in size during ontogeny, histological analysis was performed. It revealed that spermatogenesis occurred year-round in the Sardinian population, with the smallest mature individual at 18.3 mm carapace length (CL). All individuals above 23.5 mm CL had testes and vasa deferentia filled with spermatozoa and were considered physiologically mature. Morphometric maturity was observed between 27.7 and 36.2 mm CL, based on appendix masculina and petasma measurements. Despite a close association, secondary sexual characteristic development and physiological maturity were not synchronized, as seen in other crustaceans. This discrepancy likely reflects differences in male reproductive strategies, as males may produce spermatophores at smaller sizes (e.g., 23.5 mm CL), though there is no evidence that such males can successfully inseminate females in the wild. Full article

(This article belongs to the Section Aquatic Animals)

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9 pages, 234 KB

Open AccessReview

Cobalt and Vitamin B12 in Dairy Cattle Nutrition: Requirements, Functions, and Interactions

by Martha Siregar, Gerald Salas-Solis and Antonio P. Faciola

Animals 2025, 15(23), 3477; https://doi.org/10.3390/ani15233477 - 2 Dec 2025

Cited by 1 | Viewed by 2417

Abstract

Minerals are essential for ruminant health, productivity, and metabolic function, with trace minerals playing critical roles at narrow dietary margins. Cobalt (Co) is essential as it supports ruminal microbial synthesis of vitamin B12 (cobalamin), which acts as a crucial cofactor in energy and [...] Read more.

Minerals are essential for ruminant health, productivity, and metabolic function, with trace minerals playing critical roles at narrow dietary margins. Cobalt (Co) is essential as it supports ruminal microbial synthesis of vitamin B12 (cobalamin), which acts as a crucial cofactor in energy and protein metabolism. This review summarizes the role of cobalt in dairy cattle nutrition, emphasizing its contribution to vitamin B12 synthesis, propionate metabolism, and milk production. Only 3–15% of dietary cobalt is converted to vitamin B12, and efficiency depends on dietary composition, forage-to-concentrate ratio, and ruminal microbiome. Deficiency leads to reduced intake, poor growth, anemia, reproductive failure, and decreased milk yield. Cow’s milk contains ~0.5 µg/L of vitamin B12, with greater concentrations in colostrum; diet composition, supplementation, and genetics contribute to variability. Current recommendations set the cobalt requirement at 0.2 mg/kg diet DM, yet multiple environmental and nutritional factors can restrict vitamin B12 synthesis. Limitations of this review include heterogeneity among the studies reviewed, such as differences in trial design, animal genetics, diet composition, and environmental conditions, which may introduce variability and affect the generalizability and consistency of the findings. Collectively, findings highlight cobalt’s pivotal role in supporting microbial activity, energy metabolism, and production outcomes in dairy cows. Full article

(This article belongs to the Section Animal Nutrition)

15 pages, 249 KB

Open AccessArticle

Attitudes Toward In-Training Assistance Dogs in University Classrooms

by Lindsey Person and Greg C. Elvers

Animals 2025, 15(23), 3476; https://doi.org/10.3390/ani15233476 - 2 Dec 2025

Viewed by 625

Abstract

Assistance dogs provide many important benefits and improve the quality of life of the humans they serve. Puppy raisers provide the initial training of dogs that may become assistance dogs—training of basic commands such as sit, socializing and desensitizing the dog by exposing [...] Read more.

Assistance dogs provide many important benefits and improve the quality of life of the humans they serve. Puppy raisers provide the initial training of dogs that may become assistance dogs—training of basic commands such as sit, socializing and desensitizing the dog by exposing the dog to different people, novel environments and novel situations. When university students act as puppy raisers, they often bring their dog with them to class. Having an in-training assistance dog in the classroom might be disruptive to some students such as those who are allergic to dogs, anxious around dogs, or who object to dogs for religious reasons. Bringing an in-training assistance dog to class might also be disruptive to the puppy raiser if they need to direct their attention away from class to the dog to manage the dog’s behavior. The in-training assistance dog itself may experience stress from being in a novel environment with novel people. This study surveyed instructors, students, and puppy raisers in classes visited by in-training assistance dogs. In general, students reported having positive attitudes about having in-training assistance dogs in class; puppy raisers reported few problems and that their dogs experience few, if any, signs of stress. The results suggest that university students can continue to provide the initial training for dogs without negative consequences to the puppy raiser, instructor, or other students in the classes visited by the in-training assistance dogs. Full article

(This article belongs to the Section Companion Animals)

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Animals, Volume 15, Issue 23 (December-1 2025) – 148 articles

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