Animals

27 pages, 1786 KB

Open AccessArticle

Sequential Galacto- and Xylo-Oligosaccharide Feeding Transiently Modulates Gut Microbiota and Upregulates Intestinal Alkaline Phosphatase in Weaning Piglets

by James S. Stanley, Stephen C. Mansbridge, Michael R. Bedford, Ian F. Connerton and Kenneth H. Mellits

Animals 2025, 15(21), 3210; https://doi.org/10.3390/ani15213210 (registering DOI) - 4 Nov 2025

Abstract

Improving growth and health at weaning remains a priority in pig production. This study investigates whether supplementation with galacto-oligosaccharides (GOSs) followed by xylo-oligosaccharides (XOSs) improves performance and gut health of healthy 28-day old weaning piglets. Pigs received either a control basal (CON) diet, [...] Read more.

Improving growth and health at weaning remains a priority in pig production. This study investigates whether supplementation with galacto-oligosaccharides (GOSs) followed by xylo-oligosaccharides (XOSs) improves performance and gut health of healthy 28-day old weaning piglets. Pigs received either a control basal (CON) diet, the CON diet containing 1% GOS for 7 days followed by the CON diet containing 0.017% XOS for 47 days (GXOS), or the CON diet for 7 days followed by the CON diet containing 0.017% XOS for 47 days (XOS). Body weight, average daily gain, average daily feed intake, and feed conversion ratio did not differ between diets from day 1 of weaning (d1) to d54. At d7, GXOS pigs showed increased jejunal and caecal α-diversity (Shannon, inverse Simpson), distinct ileal β-diversity (Yu and Clayton, Bray–Curtis, Jaccard), and greater short-chain fatty acid-producing Lactobacillus and Veillonella; no taxa remained differentially abundant by d22, and the XOS group showed no microbiota shifts throughout the study. Jejunal goblet cell density was lower in GXOS pigs at d7. Jejunal and caecal IL-1β, IL-6, IL-8, and IL-10 gene expression was transiently greater at d7 in GXOS pigs, whereas by d22 cytokine/chemokine differences resolved, whilst intestinal alkaline phosphatase was upregulated in the ileum and caecum (XOS) and colon (GXOS and XOS). Sequential prebiotic switching and delayed XOS onset likely missed the immediate post-weaning window, during which the gut microbiota is most receptive to dietary modulation, consequently limiting potential performance gains; therefore, prebiotic timing, sequence, and duration are critical to achieving functional benefits at weaning. Full article

(This article belongs to the Special Issue Functional Prebiotics and Probiotics: Impact on Animal Growth, Development and Health)

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47 pages, 1674 KB

Open AccessReview

The Influence of Different Light Spectra on Broiler Chicken Endocrine Systems and Productivity

by Lenuța Galan, Gheorghe Solcan and Carmen Solcan

Animals 2025, 15(21), 3209; https://doi.org/10.3390/ani15213209 (registering DOI) - 4 Nov 2025

Abstract

In birds, light can penetrate the cranial bones and reach deep brain regions, where non-visual photoreceptors, especially in the hypothalamus, detect spectral and photoperiodic cues. Alongside retinal photoreception, deep-brain light sensing contributes to circadian entrainment and regulates melatonin secretion by the pineal gland. [...] Read more.

In birds, light can penetrate the cranial bones and reach deep brain regions, where non-visual photoreceptors, especially in the hypothalamus, detect spectral and photoperiodic cues. Alongside retinal photoreception, deep-brain light sensing contributes to circadian entrainment and regulates melatonin secretion by the pineal gland. These light-driven pathways modulate endocrine activity, playing a key role in muscle development. This review explores how monochromatic light-emitting diode (LED) illumination, particularly green and blue wavelengths, affects the somatotropic axis (growth hormone-releasing hormone [GHRH]-growth hormone [GH]-insulin-like growth factor 1 [IGF-1]), the gonadal axis (gonadotropin-releasing hormone [GnRH]-luteinizing hormone [LH]/follicle-stimulating hormone [FSH]-sex steroids [testosterone, estrogen, progesterone]), the thyroid axis (thyrotropin-releasing hormone [TRH]-thyroid-stimulating hormone [TSH]-thyroxine [T₄]/triiodothyronine [T₃]), and the hypothalamic-pituitary-adrenal (HPA) axis (corticotropin-releasing hormone [CRH]-adrenocorticotropic hormone [ACTH]-corticosterone). Green light enhances early-stage muscle growth via GHRH and IGF-1 upregulation, while blue light supports later myogenic activity and oxidative balance. Light schedules also influence melatonin dynamics, which in turn modulate endocrine axis responsiveness to photic cues. Furthermore, variations in photoperiod and exposure to artificial lights at night (ALAN) affect thyroid activity and HPA axis reactivity, influencing metabolism, thermoregulation, and stress resilience. Together, ocular and intracranial photoreception form a complex network that links environmental light to hormonal regulation and muscle growth. These insights support the strategic use of LED lighting to optimize broiler performance and welfare. Full article

(This article belongs to the Special Issue Enhancing Sustainable Poultry Production: Innovations in Management and Nutrition)

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14 pages, 1154 KB

Open AccessArticle

Birds as Environmental Bioindicators of Genotoxicity in Brazilian Cerrado Farmlands: An In Situ Approach

by Henrique Nazareth Souto, Edimar Olegário de Campos Júnior, Marcos Vinicius Bohrer Monteiro Siqueira, Carlos Fernando Campos, Cassio Resende Morais, Boscolli Barbosa Pereira and Sandra Morelli

Animals 2025, 15(21), 3208; https://doi.org/10.3390/ani15213208 (registering DOI) - 4 Nov 2025

Abstract

Birds have played a crucial role as environmental monitors throughout history, ranging from the use of canaries to detect methane and carbon monoxide in mines to the decline of raptors and seabirds during the DDT era due to widespread organochlorine pesticide contamination. Owing [...] Read more.

Birds have played a crucial role as environmental monitors throughout history, ranging from the use of canaries to detect methane and carbon monoxide in mines to the decline of raptors and seabirds during the DDT era due to widespread organochlorine pesticide contamination. Owing to their high diversity and capacity for bioaccumulation, birds are widely recognized as effective indicators of environmental change and pollutant exposure. Cytogenetic techniques have been increasingly applied over the past two decades to assess micronuclei formation resulting from interactions with clastogenic and aneugenic chemical compounds. The main goals of this study were (a) to evaluate a subset of the bird community in the southeastern Brazilian Cerrado as potential environmental indicators of pesticide exposure using the erythrocyte micronucleus test and (b) to investigate possible associations between bird morphometric traits and micronuclei frequency. Birds were sampled from three groups of coffee farms in the Brazilian Cerrado. Blood samples were collected from 152 individuals (122 on farms and 30 at the reference site) via the metatarsal vein, followed by slide preparation for micronucleus analysis. Two slides were prepared per bird; each slide was scored for 10,000 erythrocytes, and MN frequency was reported as the mean across slides. The species Leptotila rufaxilla, Volatinia jacarina, Galbula ruficauda, Gnorimopsar chopi, Molothrus bonariensis, Passer domesticus, Turdus leucomelas, and Turdus rufiventris exhibited six or more micronuclei per 10,000 erythrocytes, indicating the highest potential as bioindicators of environmental contamination. Micronuclei frequency in erythrocytes was positively correlated with the use of mixed pesticides, with variation depending on the size of the coffee farms. Although a slight negative biological trend was observed between micronuclei frequency and certain morphometric traits, particularly bill length, no statistically significant correlations were found. Similarly, birds from large farms exhibited a slight reduction in certain morphometric features, though these differences were also not statistically significant. These results highlight the utility of selected bird species as early-warning bioindicators for pesticide exposure in tropical agroecosystems. Full article

(This article belongs to the Topic Ecology, Management and Conservation of Vertebrates: 2nd Edition)

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Graphical abstract

15 pages, 2358 KB

Open AccessArticle

Protective Effect of Nanobodies Targeting Sip Protein Against Streptococcus agalactiae Infection in Tilapia (Oreochromis niloticus)

by Zhishen Wang, Huiling Wu, Weihao He, Shunqiang Wei, Xuemin Wei, Chaoshuai Wei, Yinghui Wang and Aiguo Huang

Animals 2025, 15(21), 3207; https://doi.org/10.3390/ani15213207 - 4 Nov 2025

Abstract

Streptococcus agalactiae (GBS) has emerged as one of the most prevalent bacterial pathogens causing severe economic losses in tilapia aquaculture due to its highly contagious and lethal nature. Nanobodies (Nbs), characterized by their small molecular size, enhanced tissue penetration, high tolerance, and exceptional [...] Read more.

Streptococcus agalactiae (GBS) has emerged as one of the most prevalent bacterial pathogens causing severe economic losses in tilapia aquaculture due to its highly contagious and lethal nature. Nanobodies (Nbs), characterized by their small molecular size, enhanced tissue penetration, high tolerance, and exceptional antigen-binding affinity, represent a promising green alternative to conventional antibiotics. In the present study, the objective was to explore the potential of specific Nbs in the treatment of tilapia GBS disease. We first screened specific Nbs targeting the surface immunogenic (Sip) protein of GBS from a naïve phage display library, and a novel nanobody Nb30 was obtained. Nb30 was expressed in Escherichia coli and purified using the Ni-NTA Agarose column. Indirect ELISA showed that Nb30 had a high affinity against Sip and GBS in vitro. Moreover, Nb30 significantly reduced GBS colonization in the liver, spleen, and brain of GBS-infected tilapia. The survival rate in the control groups was 53%, whereas it was increased to 86% after treatment with 100 mg/kg Nb30. Transcriptome profiling revealed that Nb30 could modulate critical biological processes, including antioxidant defense, immune regulation, amino acid/protein synthesis, and energy metabolism in the liver tissues of GBS-infection tilapia. Notably, the expression levels of antioxidant enzymes (cat and gpx) were significantly up-regulated, and the TLR/MyD88/NF-κB pathway-related genes (tlr5, myd88, irak4, traf6, Rela, and NF-κB2) were significantly down-regulated after treatment with Nb30. Collectively, these findings establish a novel therapeutic strategy for controlling GBS infection in tilapia and provide evidence supporting the application of nanobodies as sustainable alternatives to antibiotics in aquaculture disease management. Full article

(This article belongs to the Special Issue Aquatic Animal Medicine and Pathology 2nd Edition: Host–Pathogen Interactions)

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13 pages, 2803 KB

Open AccessFeature PaperArticle

Time Budgets in Domesticated Male Icelandic Horses on Pasture Turnout in Winter and Spring

by Daisy E. F. Taylor, Bryony E. Lancaster and Andrea D. Ellis

Animals 2025, 15(21), 3206; https://doi.org/10.3390/ani15213206 - 4 Nov 2025

Abstract

There are few 24 h time budgets for horses, especially for domesticated horses kept at pasture. Most time budgets utilise short-term scan sampling, which can miss behaviours. This study aimed to assess the seasonal variation in continuous behaviour of domesticated Icelandic horses at [...] Read more.

There are few 24 h time budgets for horses, especially for domesticated horses kept at pasture. Most time budgets utilise short-term scan sampling, which can miss behaviours. This study aimed to assess the seasonal variation in continuous behaviour of domesticated Icelandic horses at pasture during winter and spring in fair weather. Eight Icelandic horses (11.25 ± 9.19 years; 7 geldings, 1 stallion) were observed in a 26 acre field. Herd location and individual behaviour were continuously observed during 3 h periods amounting to 3 × 24 h in winter and late spring, compiled over 43 days (~21 days per season). Seasonal variation in behaviour (ANOVA), body condition (RMANOVA), and age-group variation (independent t-test) were assessed, as well as associations between weather, time period, and habitat choice (chi-square). During spring, horses showed more foraging (+18%; p < 0.001), movement (+0.5%; p < 0.05), recumbency (+5.7%; p < 0.01) and less standing (−24.6%; p < 0.001) than in winter. Behavioural synchronicity occurred between adult and juvenile horses. Mean body condition reduced from 5.6 to 4.8 in the winter. Habitat preferences varied by daytime and season, and non-feeding periods lasted less than 2 h. The 24 h foraging activity (winter: 12.7 ± 0.4 h, spring: 17 ± 0.25 h) supported the current recommendation of 12 h/24 h for domesticated horses to meet ethological requirements. Full article

(This article belongs to the Section Equids)

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16 pages, 527 KB

Open AccessFeature PaperArticle

Hock Lesions, Cow Hygiene, and Compost Quality in Compost-Bedded Pack Barns in Germany

by Phillip Andreas Guhl, Lisa Bachmann and Maike Heppelmann

Animals 2025, 15(21), 3205; https://doi.org/10.3390/ani15213205 - 4 Nov 2025

Abstract

The aim of the study was to evaluate the prevalence of hock lesions and cow hygiene as a cross-sectional study in dairy cows housed in compost-bedded pack barns (CBPs) in southern Germany. The effects of season, compost variables, and housing conditions on cow [...] Read more.

The aim of the study was to evaluate the prevalence of hock lesions and cow hygiene as a cross-sectional study in dairy cows housed in compost-bedded pack barns (CBPs) in southern Germany. The effects of season, compost variables, and housing conditions on cow hygiene and hock lesion prevalences were also investigated. Eight farms that housed their cows in CBPs were visited once in the cold season and once in the warm season between January and December 2023. All cows (cold season n = 592; warm season n = 613) were scored for hygiene and hock lesions at each visit. Compost samples were collected for laboratory analysis, and the quality of the compost-bedded pack and condition of the lying surface and concrete walkways were assessed. The udder was the cleanest body zone in both seasons; poor udder hygiene (too dirty score) occurred in 15.0% of cows in the cold season and 7.5% in the warm season (p ≤ 0.05). Only 1% of the cows had a hairless area on a hock in the cold season compared with 3.8% in the warm season; 0.2% of the cows also had swelling of the hock in the warm season (p ≤ 0.05). The compost variables that impacted cow hygiene most frequently were dry matter and compost temperature. Based on our results, CBPs reduce the prevalence of hock lesions. Cow hygiene was affected by various factors, and therefore good management of CBPs is required for good cow hygiene. Full article

(This article belongs to the Section Cattle)

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26 pages, 4873 KB

Open AccessArticle

Evaluating the Effects of Managed Free-Roaming Cat Populations on Prey Through Stable Isotope Analysis: A Pilot Study from British Columbia, Canada

by Valentina Martinoia, Renee Ferguson, Peter J. Wolf, Mario Carić, Mario Novak and Shelly Roche

Animals 2025, 15(21), 3204; https://doi.org/10.3390/ani15213204 - 4 Nov 2025

Abstract

Free-roaming domestic cats (Felis catus) present a major management challenge for animal welfare and biodiversity conservation. Trap-neuter-return (TNR) programs, which include sterilization and return of cats, are increasingly adopted to manage cat populations, often alongside routine food provisioning. However, their effectiveness [...] Read more.

Free-roaming domestic cats (Felis catus) present a major management challenge for animal welfare and biodiversity conservation. Trap-neuter-return (TNR) programs, which include sterilization and return of cats, are increasingly adopted to manage cat populations, often alongside routine food provisioning. However, their effectiveness in reducing cats’ reliance on wild prey remains contested. In this study, we use stable isotope analysis (δ¹³C, δ¹⁵N, δ³⁴S) of cat fur to investigate dietary patterns before and after TNR implementation in the context of concurrent changes in food availability linked to the closure of nearby mink-farming operations. We analyzed samples from 122 cats in a large-scale TNR initiative on a rural property in British Columbia, Canada. These included indoor cats (control), free-roaming cats prior to TNR (Group 1), a subset of Group 1 re-sampled months after food provisioning began (Run 2), and newly sampled cats that had been fed regularly before trapping (Group 2). Local prey and food sources were also analyzed to provide a comparative isotopic baseline. Our results show clear dietary shifts following TNR. Group 1 cats exhibited high isotopic variability and elevated δ¹⁵N and δ³⁴S values, consistent with wild prey consumption. In contrast, post-TNR cats showed significantly lower and more homogeneous values, aligning closely with those of indoor, kibble-fed cats. These changes are consistent with a reduced dietary reliance on wildlife and raw mink feed following the combination of TNR with regular provisioning and the cessation of mink operations. These findings demonstrate that regular food provisioning in TNR-managed colonies, particularly when combined with broader environmental changes, can significantly alter cat diets and potentially reduce their dependence on wild prey. Full article

(This article belongs to the Section Animal Welfare)

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15 pages, 1643 KB

Open AccessArticle

Assessment of Habitat Suitability for Amphioxus in the Changli Marine Reserve and Adjacent Coastal Waters, Hebei Province

by Yongfeng Zhang, Qiuzhen Wang, Quanying Wang, Qianqian Zhao, Weijie Shi, Yong Zhang, Yuan Yao and Jianle Zhang

Animals 2025, 15(21), 3203; https://doi.org/10.3390/ani15213203 - 3 Nov 2025

Abstract

Amphioxus, a key model organism in vertebrate evolution, is essential for understanding ecological balance and species diversity. This study examined the spatiotemporal distribution of Branchiostoma japonicum and assessed its habitat suitability in the Changli Golden Coast Nature Reserve and adjacent coastal waters from [...] Read more.

Amphioxus, a key model organism in vertebrate evolution, is essential for understanding ecological balance and species diversity. This study examined the spatiotemporal distribution of Branchiostoma japonicum and assessed its habitat suitability in the Changli Golden Coast Nature Reserve and adjacent coastal waters from 2008 to 2023 (excluding 2020). The maximum abundance showed marked fluctuations, with a sharp decline between 2008 and 2015 followed by recovery after 2016, reaching a peak of 345 ind./m² in 2022. The average abundance also increased, peaking at 34 ind./m² in 2022. Spatially, the higher abundances occurred in central stations, while peripheral sites were much lower, sometimes absent. Spearman’s correlation and Principal Component Analysis (PCA) identified sediment grain size (1~0.25 mm), water depth and sediment sulfide as key environmental factors. A habitat suitability index (HSI), constructed using the Analytic Hierarchy Process (AHP), showed higher values in central stations, indicating more favorable conditions. These findings highlight the importance of long-term monitoring, clarify the ecological requirements of amphioxus, and provide guidance for habitat conservation and management in regions affected by environmental change and human activities. Full article

(This article belongs to the Section Aquatic Animals)

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18 pages, 1279 KB

Open AccessArticle

Integrative Multivariate Analysis of Milk Biomarkers, Productive Performance, and Animal Welfare Indicators in Dairy Cows

by Daniela Elena Babiciu, Florin Ioan Beteg, Mihai Cenariu, Anamaria Blaga Petrean, Sorin Marian Mârza, Eva Andrea Lazar and Silvana Popescu

Animals 2025, 15(21), 3202; https://doi.org/10.3390/ani15213202 - 3 Nov 2025

Abstract

Animal welfare is increasingly recognised as a core component of sustainable dairy production, yet objective assessment at the herd level remains challenging. This study evaluated whether milk biomarkers can serve as non-invasive indicators of cow welfare. Thirty-seven dairy farms were assessed using the [...] Read more.

Animal welfare is increasingly recognised as a core component of sustainable dairy production, yet objective assessment at the herd level remains challenging. This study evaluated whether milk biomarkers can serve as non-invasive indicators of cow welfare. Thirty-seven dairy farms were assessed using the Welfare Quality^® protocol and various milk analysis parameters. As a first line of results, Spearman correlations revealed strong associations between milk biomarkers and welfare indicators. For example, a higher fat-to-protein ratio was linked to better feeding, lower prevalence of hunger, and improved human–animal relationships. In contrast, elevated somatic cell count and differential somatic cell count were associated with mastitis, lameness, dirtiness, and reduced emotional well-being. Using Principal Component Analysis (PCA), three dimensions were identified, health–hygiene, socio-behavioural, and metabolic stress, explaining 44.7% of variance. K-means clustering distinguished three herd profiles: feeding–metabolic balance, behavioural–comfort, and clinical–hygiene risk. These findings demonstrated that routine milk biomarkers provide integrated, non-invasive information on herd health, behaviour and, comfort. Incorporating routine milk analysis into welfare assessments can support the early detection of issues, facilitate evidence-based decision-making, and promote sustainable dairy management. Full article

(This article belongs to the Special Issue Livestock Welfare in Extensive Production System)

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18 pages, 2442 KB

Open AccessArticle

Integrative Analysis of Gene Networks Associated with Adipose and Muscle Traits in Hanwoo Steers

by Suk Hwang, Taejoon Jeong, Junyoung Lee, Woncheoul Park, Sunsik Jang and Dajeong Lim

Animals 2025, 15(21), 3201; https://doi.org/10.3390/ani15213201 - 3 Nov 2025

Abstract

This study aims to characterize tissue-specific expression patterns in Hanwoo steers by identifying co-expression modules, functional pathways, and hub genes related to fat and muscle traits using Weighted Gene Co-expression Network analysis (WGCNA). RNA-Seq data were generated from three muscle tissues (longissimus muscle, [...] Read more.

This study aims to characterize tissue-specific expression patterns in Hanwoo steers by identifying co-expression modules, functional pathways, and hub genes related to fat and muscle traits using Weighted Gene Co-expression Network analysis (WGCNA). RNA-Seq data were generated from three muscle tissues (longissimus muscle, tenderloin, and rump) and two fat tissues (back fat and abdominal fat) collected from six 30-month-old Hanwoo steers. Quality control of raw sequencing reads was performed using FastQC, and trimmed reads were aligned to the bovine reference genome (ARS-UCD1.3) using HISAT2. We also identified a gene co-expression network via WGCNA using normalized gene expression values. Modules were defined based on topological overlap and correlated with tissue-specific expression patterns. Modules with a significant association (p < 0.05) were used for functional enrichment based on Gene Ontology (GO) and KEGG pathways, as well as Protein–Protein Interaction Network analysis. A total of seven co-expression modules were identified by WGCNA and labeled in distinct colors (yellow, blue, red, brown, turquoise, green, black). Among them, the yellow and blue modules were positively associated with back fat, while the turquoise and green modules showed a negative correlation with abdominal fat. Additionally, the turquoise or green module was positively correlated with longissimus and rump tissues, indicating distinct gene expression patterns between fat and muscle. This study identified key co-expression modules and hub genes associated with muscle and fat metabolism. Notably, ARPC5 (blue module) was involved in lipid metabolism and energy storage, whereas AGPAT5 (turquoise module) was linked to maintaining muscle cell structure and function. These findings reveal biological mechanisms for tissue-specific gene regulation, providing targets for enhancing meat quality in Hanwoo. Full article

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

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17 pages, 283 KB

Open AccessFeature PaperArticle

Growth Performance, Carcass and Meat Quality Traits of Three Rabbit Lines Under Heat Stress Conditions

by Emanuele Pontalti, Zsolt Matics, Marco Cullere, Zsolt Szendrő, Zsolt Gerencsér, Bianca Palumbo and Antonella Dalle Zotte

Animals 2025, 15(21), 3200; https://doi.org/10.3390/ani15213200 - 3 Nov 2025

Abstract

Given the yearly challenging environmental scenario with more and more frequent and intense heat waves, the livestock sector has to find affordable and sustainable solutions to face the expected increase in meat demand by 2050. Among livestock species, rabbits are particularly sensitive to [...] Read more.

Given the yearly challenging environmental scenario with more and more frequent and intense heat waves, the livestock sector has to find affordable and sustainable solutions to face the expected increase in meat demand by 2050. Among livestock species, rabbits are particularly sensitive to heat stress (HS) but, paradoxically, the scientific background on the response of different genetics to environmental stressors like HS is rather scarce. This is a significant gap, especially considering that most of the demographic growth, and meat demand, is expected in developing countries where rabbits play a key role in subsistence farming. Therefore, this research investigated the effects of environmental temperature (Control—20 °C; High—28 °C) on growth performance, slaughter traits and meat quality of three Hungarian rabbit genotypes (Pannon Large—PL; Pannon White—PW; Pannon Ka—PK). Animals (n = 360) were housed in wire-mesh cages (3 animals/cage) in two separate controlled-temperature rooms (60 rabbits/genotype/room), from 5 to 11 weeks of age, during which they received ad libitum feed and water. Even if the three genotypes were exposed to the same environmental challenge, they exhibited different responses. The PL line showed superior performance, with the highest carcass weight and yield (p < 0.001), and the greatest water-holding capacity (p < 0.01) in the loin muscle. The PW rabbits showed the largest reduction in overall weight gain (−24.7%; p < 0.001) and the lowest decrease in feed conversion ratio (−3.20%; p < 0.001). PK rabbits experienced the greatest reduction in total dissectible fat (−34.6%; p < 0.001) and hind leg lipid content (−20.3%; p < 0.01), with the highest proportion of polyunsaturated fatty acids (p < 0.01), which fostered meat lipid oxidation (p < 0.05). As expected, these differences in performance and meat quality traits reflected the distinct selection criteria and genetic background of these genotypes: the PL is a paternal line, the PK is a maternal line, and the PW is a productive line. Regarding the temperature effect, PK and PW genotypes were the most impacted by chronic HS: PW rabbits suffered the largest performance depression, while PK rabbits showed the worst carcass and meat quality traits. Instead, PL rabbits demonstrated the best outcomes under chronic HS, showing the greatest productive efficiency and satisfactory meat quality traits. Full article

(This article belongs to the Special Issue Impact of Climate Change on Animal Welfare, Health, and/or Product Quality of Livestock Species)

18 pages, 2718 KB

Open AccessArticle

Multi-Omics Insights into the Impact of Fermented Wheat Bran-Soybean Meal-Broussonetia papyrifera Mixture Substance on the Gut Microbiota of Late Gestation Sows In Vitro

by Lele Fu, Yushi Chen, Yantao Li and Cheng Wang

Animals 2025, 15(21), 3199; https://doi.org/10.3390/ani15213199 - 3 Nov 2025

Abstract

Comprehensive maternal nutritional interventions, particularly during late gestation, enhance perinatal outcomes and support long-term maternal-offspring health by modulating the microbiota. Fermented diets are recommended for inclusion in dietary guidelines during gestation, yet the specific metabolites after fermentation and their specific regulatory effects on [...] Read more.

Comprehensive maternal nutritional interventions, particularly during late gestation, enhance perinatal outcomes and support long-term maternal-offspring health by modulating the microbiota. Fermented diets are recommended for inclusion in dietary guidelines during gestation, yet the specific metabolites after fermentation and their specific regulatory effects on gut microbiota during late gestation remain unclear. This study investigates the functional benefits of a fermented wheat bran–soybean meal–Broussonetia papyrifera mixed substrate (FMS) on the late-gestation gut microbiota using an in vitro fermentation model. The FMS was first fermented for 72 h with bacterial and enzymatic agents (2% v/v), then anaerobically incubated with fecal inocula from Jinhua pigs. Fermentation significantly enhanced nutritional profiles, increasing crude protein and amino acids while reducing fiber components (neutral detergent fiber, acid detergent fiber, and non-starch polysaccharide, p < 0.05). Metabolome analysis revealed a significant increase in the abundance of organic acids, amino acids, and short peptides in FMS, along with the enrichment of D-amino acid and sphingolipid pathways (p < 0.05). In addition, FMS significantly increased the abundance of Limosilactobacillus and Lactobacillus, as well as short-chain fatty acids production, compared to the unfermented group (p < 0.05). These findings demonstrate that fermentation pretreatment reduces fiber components, enhances flavor compounds and bioactive metabolites, thereby optimizing microbial utilization and increasing short-chain fatty acids production. Full article

(This article belongs to the Collection The Weaned Pig: Nutrition and Management)

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15 pages, 976 KB

Open AccessArticle

Seasonal Trends in Cardiac Troponin I Concentration and Creatine Kinase and Aspartate Aminotransferase Enzyme Activity in Relation to Myocardial Velocity Rates in Eventing Horses

by Insa Rump-Dierig, Johanna Giers, Charlotte Frenzel, Sabita Stöckle and Heidrun Gehlen

Animals 2025, 15(21), 3198; https://doi.org/10.3390/ani15213198 - 3 Nov 2025

Abstract

This study examines seasonal changes in muscle and heart parameters in eventing horses over the course of a competition season. Blood levels of the enzymes creatine kinase (CK) and aspartate aminotransferase (AST) as well as the heart muscle-specific concentration of cardiac troponin I [...] Read more.

This study examines seasonal changes in muscle and heart parameters in eventing horses over the course of a competition season. Blood levels of the enzymes creatine kinase (CK) and aspartate aminotransferase (AST) as well as the heart muscle-specific concentration of cardiac troponin I (cTnI) were measured before (pre), 30 min (p30) and 24 h (p24) after competitions. Creatine kinase (CK: median pre-competition = 175 U/L, 30 min post = 221 U/L, 24 h post = 140 U/L), aspartate aminotransferase (AST: pre = 319 U/L, p30 = 335 U/L, p24 = 333 U/L), and cardiac troponin I concentration (cTnI: pre = 0.006 ng/mL, p30 = 0.011 ng/mL, p24 = 0.007 ng/mL) exhibited partial normalization by 24 h post-exercise but at the same time demonstrated significant seasonal variation (p < 0.001). Echocardiographic assessments revealed sustained high-level myocardial velocities, with occasional modest seasonal declines. A significant correlation was identified between cTnI levels and early diastolic myocardial velocity (Em) (Spearman’s Rho: pre-exercise 0.323, Rho p30: 0.357), whereas a negative correlation was manifest at 24 h (Spearman’s Rho = −0.300). These findings suggest a heightened sensitivity of diastolic myocardial velocity to myocardial injury. Given that diastolic dysfunction frequently constitutes an early manifestation of myocardial compromise, our results underscore the utility of biomarkers alongside myocardial velocity measures as valuable tools for the early detection of subclinical fatigue in high-performance sport horses. Full article

(This article belongs to the Special Issue Advances in Internal Medicine in Equids)

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16 pages, 4454 KB

Open AccessArticle

A New Species of Boulenophrys (Anura, Megophryidae) from Northern Jiangxi, China

by Deming Shen, Haiying Zhou, Kevin R. Messenger, Hina Amin, Zhenyu Wang, Jun Xu, Shi Xu and Yankuo Li

Animals 2025, 15(21), 3197; https://doi.org/10.3390/ani15213197 - 3 Nov 2025

Abstract

A new species of the genus Boulenophrys (Anura, Megophryidae) is described from Mount Lushan, Jiangxi Province, China, long considered to be Boulenophrys boettgeri. Phylogenetic analyses based on mitochondrial COI and 16S rRNA genes show that the new species forms a sister clade [...] Read more.

A new species of the genus Boulenophrys (Anura, Megophryidae) is described from Mount Lushan, Jiangxi Province, China, long considered to be Boulenophrys boettgeri. Phylogenetic analyses based on mitochondrial COI and 16S rRNA genes show that the new species forms a sister clade to B. jinggangensis, with an uncorrected COI p-distance of 3.7%, confirming its status as a distinct species. Morphologically, it differs from all known congeners by a unique combination of characters, and from its closest relative B. jinggangensis by having the following traits: (1) larger adult body size (adult males SVL 42.7–44.7 mm; adult females 46.0–50.3 mm); (2) heels not meeting when hindlimbs are flexed at right angles to the body axis; (3) smaller horn-like tubercle on the upper eyelid; and (4) relative finger lengths (I < II < IV < III). Acoustically, the new species differs markedly from B. jinggangensis in nearly all parameters of its advertisement calls. This discovery increases the known species diversity of Boulenophrys in Jiangxi Province and provides baseline data for further biodiversity conservation efforts in the Mount Lushan region. Full article

(This article belongs to the Special Issue Evolution, Diversity, and Conservation of Herpetofauna: Second Edition)

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10 pages, 223 KB

Open AccessCase Report

Salmonellosis Outbreak in a Rottweiler Kennel Associated with Raw Meat-Based Diets

by Betina Boneva-Marutsova, Plamen Marutsov, Marie-Louise Geisler and Georgi Zhelev

Animals 2025, 15(21), 3196; https://doi.org/10.3390/ani15213196 - 3 Nov 2025

Abstract

This case describes an outbreak of salmonellosis caused by Salmonella enterica subspecies enterica serotype Agona in a Rottweiler breeding kennel, associated with raw meat-based diet (RMBD) of unlicensed origin. The report presents the clinical, epidemiological, and microbiological characteristics of the outbreak, as well [...] Read more.

This case describes an outbreak of salmonellosis caused by Salmonella enterica subspecies enterica serotype Agona in a Rottweiler breeding kennel, associated with raw meat-based diet (RMBD) of unlicensed origin. The report presents the clinical, epidemiological, and microbiological characteristics of the outbreak, as well as the control and preventive measures undertaken. Methods: Samples of faeces, vomit, raw food, and environmental surfaces were collected and examined. The isolated pathogen was identified using bacteriological culture, biochemical testing, MALDI-TOF mass spectrometry, and serotyping according to the White–Kauffmann–Le Minor scheme. Antimicrobial susceptibility was determined by the broth microdilution method in accordance with standards of Clinical and Laboratory Standards Institute (CLSI). Results: Clinical signs included vomiting, diarrhoea, lethargy, and dehydration without fever, with disease exacerbation observed in post-partum animals. Extensive carriage and faecal shedding of S. Agona were detected in affected dogs, along with widespread contamination of food and the kennel environment. The isolate was susceptible to some antimicrobial agents but resistant to cephalexin, aminoglycosides, lincosamides, macrolides, and fusidic acid, and showed intermediate susceptibility to polymyxin B. Following discontinuation of raw meat feeding, targeted antimicrobial therapy, and environmental disinfection, all dogs recovered, and subsequent tests for Salmonella spp., were negative. All human contacts also tested negative. Conclusions: This represents the first documented outbreak of S. Agona infection in dogs in Bulgaria linked to a RMBD. The findings emphasise the importance of feed safety, biosecurity, and traceability of feed sources in kennels, as well as the potential zoonotic risk associated with raw feeding practices. The diagnostic and therapeutic measures implemented in this case provide an effective model for managing similar epidemiological events within the One Health framework. Full article

(This article belongs to the Section Veterinary Clinical Studies)

12 pages, 375 KB

Open AccessArticle

Free Plasma Amino Acid Concentrations in Horses Fed Different Dosing Regimens of Hydrolysed Collagen

by Lieuwke C. Kranenburg, Katharina S. Reinke, Jan van den Broek, Esther A. Zaal, Robin van den Boom and David A. van Doorn

Animals 2025, 15(21), 3195; https://doi.org/10.3390/ani15213195 - 3 Nov 2025

Abstract

Hydrolysed collagen is used as a supplement for horses with osteoarthritis, hoof horn growth problems, and gastric ulcers. To determine the oral availability of a specific hydrolysed collagen supplement and the appropriate dose, six Warmblood mares were fed two different concentrations of the [...] Read more.

Hydrolysed collagen is used as a supplement for horses with osteoarthritis, hoof horn growth problems, and gastric ulcers. To determine the oral availability of a specific hydrolysed collagen supplement and the appropriate dose, six Warmblood mares were fed two different concentrations of the supplement: 100 g HC (C_H), 50 g HC (C_L), and a control of 0 g HC (C_N) during one week in a randomised cross-over design. On day 7, 14 and 21, blood sampling for amino acid (AA) analysis was performed, just prior to feeding the supplement (t = 0) and every hour after feeding for 8 h (t = 1–8). Statistical analysis revealed differences in mean plasma AA concentrations between the CH and CN doses for alanine, arginine, glutamine, glycine, proline, serine and hydroxyproline. Similarly, statistical differences were observed between the CL and CN doses for arginine, glycine, proline and hydroxyproline. This study demonstrated the availability of amino acids from the supplemented hydrolysed collagen. Although clinical efficacy was not evaluated in this study, a dose of 100 g HC once daily resulted in higher plasma concentrations, which remained detectable for at least 24 h, suggesting greater clinical relevance. Full article

(This article belongs to the Section Equids)

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22 pages, 1443 KB

Open AccessArticle

CNN-Based Identification of Pathogens of Concern in Shrimp

by Tharyar Aung, Rapeepun Vanichviriyakit, Kittisak Chayantrakom, Somkid Amornsamankul and Pallop Huabsomboon

Animals 2025, 15(21), 3194; https://doi.org/10.3390/ani15213194 - 3 Nov 2025

Abstract

Concerning shrimp diseases, including acute hepatopancreatic necrosis disease (AHPND), hepatopancreatic parvovirus (HPV) infection and Enterocytozoon hepatopenaei (EHP) microsporidiosis negatively impact shrimp aquaculture through acute mortality, chronic growth retardation or compromised health that increases susceptibility to concurrent infections. All three diseases damage hepatopancreas, a [...] Read more.

Concerning shrimp diseases, including acute hepatopancreatic necrosis disease (AHPND), hepatopancreatic parvovirus (HPV) infection and Enterocytozoon hepatopenaei (EHP) microsporidiosis negatively impact shrimp aquaculture through acute mortality, chronic growth retardation or compromised health that increases susceptibility to concurrent infections. All three diseases damage hepatopancreas, a vital organ for nutrient absorption and growth, though their clinical outcomes differ: AHPND is typically associated with rapid, high mortality, EHP primarily causes chronic production losses and HPV, while currently of lower pathogenic significance, may still impair health under certain conditions. Outbreak severity is often intensified by poor water quality, inadequate farm management, antibiotic misuse and pathogen vectors, leading to substantial economic losses. Timely and accurate diagnosis is therefore critical for effective disease management. This study investigates two convolutional neural network (CNN) architectures, EfficientNet and MobileNet. A curated and preprocessed dataset was used to fine-tune both models with a standardized custom classification head, ensuring a controlled backbone comparison. Experimental results show both architectures achieving over 95% accuracy, with MobileNet providing faster inference suitable for on-site deployment. These findings demonstrate the practical feasibility of lightweight CNN-based diagnostics tools for real-time, scalable, and cost-efficient health monitoring in shrimp aquaculture, bridging the gap between the laboratory-grade performance and field-level usability. Full article

(This article belongs to the Topic AI, Deep Learning, and Machine Learning in Veterinary Science Imaging)

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16 pages, 2043 KB

Open AccessArticle

Melatonin Rescues Triclosan-Disrupted Porcine Oocyte Meiosis via Suppression of p53-Mediated Apoptosis

by Jiaxin Duan, Ning Zhao, Shibin Wang, Xinyu Li, Bugao Li and Guoqing Cao

Animals 2025, 15(21), 3193; https://doi.org/10.3390/ani15213193 - 3 Nov 2025

Abstract

Triclosan (TCS), a widely used environmental antimicrobial agent, poses potential risks to female reproductive health, yet its toxic effects on oocyte maturation remain inadequately characterized. In this study, we established an in vitro maturation (IVM) model of porcine oocytes to investigate TCS-induced meiotic [...] Read more.

Triclosan (TCS), a widely used environmental antimicrobial agent, poses potential risks to female reproductive health, yet its toxic effects on oocyte maturation remain inadequately characterized. In this study, we established an in vitro maturation (IVM) model of porcine oocytes to investigate TCS-induced meiotic impairment and to evaluate the rescuing effects of melatonin (MT), an endogenous indoleamine with potent antioxidant and anti-apoptotic activities. Our results demonstrated that TCS exposure significantly disrupted oocyte maturation, as evidenced by suppressed polar body extrusion and compromised cumulus expansion. Furthermore, TCS triggered early apoptosis. Proteomic analysis revealed that the p53 signaling pathway was significantly dysregulated by TCS exposure. Notably, co-treatment with MT during IVM effectively restored meiotic progression, attenuated apoptosis, and rebalanced the disrupted proteomic profile. Mechanistic investigation, validated by Western blotting, confirmed that TCS upregulated p53 and downregulated its downstream cell cycle effector CCNB1 while concurrently altering the ratio of apoptosis-related proteins BAX/BCL-2. Melatonin treatment effectively normalized the expression of these key proteins (p53, CCNB1, BAX, and BCL-2). These findings illustrate that MT rescues TCS-impaired oocyte quality through p53-dependent suppression of apoptosis and restoration of meiotic progression, providing new insights into potential strategies for mitigating environmental pollutant-induced reproductive damage. Full article

(This article belongs to the Section Animal Reproduction)

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20 pages, 4147 KB

Open AccessArticle

A Patch and Attention Mechanism-Based Model for Multi-Parameter Prediction of Rabbit House Environmental Parameters

by Ronghua Ji, Guoxin Wu, Hongrui Chang, Zhongying Liu and Zhonghong Wu

Animals 2025, 15(21), 3192; https://doi.org/10.3390/ani15213192 - 2 Nov 2025

Abstract

The health and productivity of rabbits are highly sensitive to the environmental conditions within the rabbit house, particularly to fluctuations and deviations in temperature, relative humidity, and carbon dioxide (CO₂) concentration. However, owing to the thermal inertia and residual evaporation effects [...] Read more.

The health and productivity of rabbits are highly sensitive to the environmental conditions within the rabbit house, particularly to fluctuations and deviations in temperature, relative humidity, and carbon dioxide (CO₂) concentration. However, owing to the thermal inertia and residual evaporation effects inherent in ventilation and cooling systems, environmental changes often exhibit delayed responses, rendering real-time control inadequate. Accurate prediction of key environmental parameters is indispensable for formulating effective environmental control strategies, as it enables consideration of their future dynamics and thereby enhances the rationality of regulation in rabbit farming. Existing prediction models often exhibit unsatisfactory accuracy and weak generalization, which restricts the incorporation of prediction into effective environmental control strategies. To address these limitations, summer indoor and outdoor environmental data were collected from rabbit houses in Nanping, Fujian; Jiyuan, Henan; and Qingyang, Gansu, China—three climatically distinct regions—forming three datasets. Based on these datasets, a multi-parameter time-series prediction model, Patch and Cross-Attention Enhanced Transformer for Rabbit House Prediction (PatchCrossFormer-RHP), is introduced, integrating patching and attention mechanisms. The model partitions the sequences of rabbit house temperature, relative humidity, and CO₂ concentration into patches and incorporates auxiliary parameters, such as indoor air velocity and outdoor temperature and humidity, to enhance feature representation. Furthermore, it applies cross-attention with differentiated encoding to disentangle multi-parameter relationships and improve predictive performance. This study used the Fujian dataset as the primary benchmark. On this dataset, PatchCrossFormer-RHP achieved root mean square error (RMSE) values of 0.290 °C, 1.554%, and 38.837 ppm for rabbit house temperature, humidity, and CO₂ concentration, respectively, with corresponding R² values of 0.963, 0.956, and 0.838, consistently outperforming RNN, GRU, and LSTM. Transfer experiments with single- and multi-source pretraining followed by fine-tuning on Fujian demonstrated that strong cross-regional generalization can be achieved with only limited target-domain data. Full article

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

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