Animals doi: 10.3390/ani16182892

Authors: Chengjie Yin Rongzhen Zhang Zewen Feng Xiaoning Shi Renhui Li Miao Xiang He Zhang Longgen Guo

Characterizing the spatiotemporal patterns of fish communities in subtropical river-type reservoirs provides useful baseline information for fisheries stewardship and aquatic ecosystem monitoring. This work investigated fish assemblages across 18 sampling sites within Shanxi Reservoir, Zhejiang Province, using an integrative approach combining gillnet surveys, hydroacoustic detection, and habitat parameter monitoring. Sampling was conducted during two periods: October 2022 (autumn, post-flood, fattening phase) and April 2023 (spring, normal water level, spawning phase). A total of 22 species were documented in spring and 21 in autumn, with Cyprinidae species predominating. Dominant species shifted seasonally, led by Hypophthalmichthys nobilis, Culter dabryi and Hemiculter leucisculus. Planktivores and carnivores were more abundant in spring, while omnivores increased in autumn. Spatially, fish populations were mainly concentrated in the shallow riverine zone (SRZ), predominantly comprising small- to medium-sized individuals. Mantel tests and Pearson correlation analyses identified associations between fish assemblage structure in the deep lake zone (DLZ) and environmental variables including dissolved oxygen, conductivity, chlorophyll a, and phytoplankton density and biomass. This study provides preliminary baseline data on fish community dynamics in Shanxi Reservoir and offers tentative observations that may help inform future zoned monitoring and ecological restoration planning.

Animals doi: 10.3390/ani16182891

Authors: Yifeng Song Zhalaga Muqier Xiaoping An Na Liu Jingwei Qi

Heat stress is known to affect dairy cow behavior, but its effects on milking-order patterns and stabilization after herd entry in large commercial herds are not well described. We analyzed 70,491 parlor records from 484 Holstein cows milked three times daily in a free-stall herd to examine how thermal conditions shape milking-order dynamics. Daily intraday milking-order displacement was quantified based on changes in queue position between successive milkings. Cow-specific regressions of milking order on ambient temperature were used to identify clusters of thermal response, and time to reach a stable queue position was evaluated for newly introduced cows according to thermal conditions at herd entry. Results show a modest increase in intraday physical displacement from thermally comfortable to severely hot days, whereas relative displacement did not differ significantly across the four thermal categories. Cows showed consistent individual differences in how their milking position shifted with temperature that were only weakly associated with parity, days in milk, milk yield, or panting score. In an exploratory severe versus non-severe comparison, cows entering the herd under severe thermal conditions had a lower adjusted hazard of reaching the predefined stabilization criterion (HR = 0.62, 95% CI: 0.43–0.90). Collectively, these findings indicate that milking-order patterns extracted from routine parlor records are associated with behavioral responses to thermal conditions. Their management implications, including decisions related to herd entry and cooling, require further experimental evaluation.

Animals doi: 10.3390/ani16182890

Authors: Lori R. Kogan Jennifer Currin-McCulloch Sohaila Jafarian Emma Klein Kathleen Cooney

As companion animals approach end of life, their caregivers often face complex decisions that require balancing their pet’s quality of life, relief from suffering, potential medical interventions, financial and resource barriers, and the difficult question of whether and when to pursue euthanasia. The purpose of this qualitative study was to explore how companion animal caregivers experience end-of-life decision-making for their pet and the care they received from their veterinary team. The study examined the experiences of 143 pet caregivers who responded to an online survey between September and December 2025. Data collection included caregiver’s responses to four open-ended prompts inquiring about their experiences with their pet’s end-of-life. Questions explored what they regarded as the most challenging and meaningful aspects, regrets they held about their pet’s death, and advice for veterinarians about how best to support pet owners in their pet’s final moments. Data were analyzed using Braun and Clarke’s reflexive thematic analysis. Results highlighted caregivers’ distress surrounding decisions about whether and when they should pursue euthanasia. Caregivers described balancing multiple considerations in their decision-making, including their pet’s overall welfare, the needs and perspectives of family members, resource constraints, access to specialized veterinary care, and their own caregiving fatigue. Their responses demonstrated grief, regret, guilt, and meaning making. Veterinarians’ compassion, transparency, and forward-thinking enabled caregivers to plan for rituals and meaningful death experiences for their pets and their families. Suggestions for veterinary care emphasized the importance of relationship-centered care in addition to technical expertise. Veterinary end-of-life care should prioritize empathy, early and transparent communication, and opportunities for cherished time during the pet’s final moments.

Animals doi: 10.3390/ani16182889

Authors: Zachary C. Crump Anthony F. Alberto Bailee L. Brown Bryce W. Roholt Lillian L. Okamoto Ryan A. Spurling Robert E. Ward Brady M. Blackett Robert L. Harding Sanjeewa D. Ranathunga Kara J. Thornton

The objective of this research was to determine the impacts of including sprouted barley (SB) in the diets of backgrounding beef steers. Angus-influenced steers (328 ± 6.59 kg) were assigned to one of four diets (n = 15/diet): a control (CON) diet, or a diet containing 5%, 10%, or 20% SB (DM basis)—replacing corn silage and some rolled barley. Individual intake and feeding behavior were monitored using Vytelle bunks. Body weight (BW) and ruminal fluid samples were collected on d 0, 30, and 60, while dry matter intake (DMI), feeding behavior, average daily gain (ADG), and gain-to-feed ratio (G:F) were monitored for 60 d. In addition, total amount of nutrients consumed (crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), net energy for maintenance (NEm), and net energy for gain (NEg)) were also assessed. Calf BW, ADG, and G:F were not impacted by diet (p ≤ 0.43). DMI and consumption of CP, NDF, ADF, NEm and NEg were all impacted by diet × day (p ≤ 0.01). The acetate:propionate, percentage of propionate and concentration of butyrate were each impacted by diet × day (p ≤ 0.007). Collectively, these findings suggest including SB in the diet modifies nutrient intake and ruminal fermentation in backgrounding steers.

Animals doi: 10.3390/ani16182888

Authors: Qiangqiang Ma Ying Chen Hong Dong Liqin Wang Xuefeng Fu Gulimire Abudureyimu Jiaxin Zhang Wei Zhang Jinbo Tian Yangsheng Wu Jiapeng Lin

This study aimed to explore whether zona-associated residual sperm may contribute to exogenous Y-DNAY-DNA contamination and cause sex bias in PCR-based sex classification of ovine early-stage embryos, based on the amplification of the AMEL gene. Primers targeting the conserved regions of the ovine AMELX and AMELY genes were designed, and a nested PCR system using AMEL and YWHAZ genes was established. Group A comprised 17 female and 33 male embryos, corresponding to a female-to-male ratio of 0.52:1, whereas Group B comprised 23 female and 27 male embryos, corresponding to a female-to-male ratio of 0.85:1. DAPI staining revealed sperm-morphology structures or structures containing sperm-derived DNA adjacent to the zona pellucida of ovine early-stage embryos. Sequence alignment of PCR amplicons showed consistency with the AMELX and AMELY reference genes. In conclusion, these results support our research hypothesis that zona-associated residual sperm may serve as a potential source of Y-DNAY-DNA contamination and interfere with PCR-based embryonic sex determination.

Animals doi: 10.3390/ani16182887

Authors: Gergely Kiss Borbála Mózes Ágnes Sterczer Katalin Lányi Krisztián Németh Roland Psáder

Bile acid (BA) dysmetabolism is implicated in canine chronic inflammatory enteropathy (CIE), but fasting serum profiles across treatment-response groups remain poorly characterized. This exploratory observational study analyzed pretreatment serum from dogs subsequently classified as food-responsive enteropathy (FRE; n = 8) or immunosuppressant-responsive enteropathy (IRE; n = 5), with four unmatched purpose-bred Beagle controls. Targeted LC–MS/MS, performed using a published canine serum method, provided results for 15 BA species. All samples were analyzed in one batch by an analyst blinded to subsequent group assignment. Eight species quantifiable in every dog and ten derived variables were compared using Kruskal–Wallis tests with Benjamini–Hochberg correction across 18 omnibus tests. Cholic acid and unconjugated primary BA concentrations were higher in IRE than in controls and FRE. The total primary-to-secondary BA ratio showed a monotonic group ordering (control < FRE < IRE; H = 13.76, p = 0.0010, q = 0.0092; epsilon-squared = 0.840), with all Bonferroni-corrected pairwise comparisons significant. A post hoc four-scenario sensitivity analysis comprising the submitted zero-substitution analysis and three LOD/LOQ-based alternatives reproduced the same 18-test FDR-significant set and the same ratio inference. This analysis tests robustness to censored-value handling but is not independent validation. The findings suggest group-associated serum BA patterns, but the small sample, clinical heterogeneity, and unmatched controls preclude diagnostic or treatment-predictive interpretation.

Animals doi: 10.3390/ani16182886

Authors: Jun Li Zhongying Li Menghan Han Mengfan Zhao Zhuoying Li Huibin Shi Haoyuan Han Kun Liu Xing Liu Kai Quan

Milk fat, a key nutritional component of milk, not only supplies energy and immune protection for offspring but also serves as a critical quality indicator for goat milk. Sirt7, one of the least studied members of the Sirtuins family, has been shown to play a regulatory role in lipid metabolism. This study investigated the regulatory role of Sirt7 in milk fat synthesis and fatty acid composition in dairy goat mammary epithelial cells (GMECs) using a Sirt7 inhibitor (ID: 97491). GMECs were treated with a Sirt7 inhibitor (ID: 97491), and cell viability was assessed via the CCK-8 assay, which confirmed that 15 μmol/L inhibitor had no cytotoxic effects. Functional analyses using RT-qPCR, Western blot, and GC-MS revealed that Sirt7 inhibition upregulated the expression of SREBP1, CD36, ACC, SCD1, and ELOVL6 while downregulating FOXO1 and FABP1. At the protein level, the inhibitor reduced Sirt7, FOXO1. Additionally, Sirt7 inhibition increased intracellular lipid droplet and triglyceride content. GC-MS analysis showed that Sirt7 inhibition decreased C11:0; increased C16:0, C15:0, C20:0, C24:0, C16:1, and C18:1n9c; and decreased C20:5n3 (EPA). Collectively, these findings suggest that pharmacological inhibition of Sirt7 is associated with upregulation of SREBP1, disruption of lipid homeostasis, triglyceride accumulation, and altered fatty acid composition in GMECs. These results provide preliminary evidence for a potential regulatory role of Sirt7 in milk fat synthesis and lay a foundation for future mechanistic studies.

Animals doi: 10.3390/ani16182885

Authors: Ángel Sebastián Rodríguez-Pazmiño Sarita Doménica Bautista-Arcentales Miguel Angel Garcia-Bereguiain Solon Alberto Orlando

University-based animal welfare organizations are grassroots actors involved in managing free-roaming dog and cat populations, yet their role remains poorly characterized, particularly in low- and middle-income countries. This study aimed to identify and describe animal welfare organizations operating within universities in Ecuador and to analyze their potential contributions within a One Health framework. A cross-sectional survey of 63 universities was conducted using publicly available information from institutional websites and social media platforms, complemented by semi-structured interviews with representatives of identified organizations. Organizations with evidence of activity through 2026 and a focus on dogs or cats were included. A total of 14 animal welfare organizations from 12 universities were identified, representing 19.0% of the universities analyzed. Most organizations were located in public universities (75.0%), and 28.6% were associated with veterinary-related programs. Thematic analysis of interview data (12 out of 14 organizations interviewed) revealed four key dimensions of organizational functioning: capacity and governance, animal care and population management, public health and community engagement, and institutional integration. Reported activities included animal rescue, sterilization campaigns, adoption initiatives, and community outreach, aligning with internationally recommended strategies for humane population management. Several organizations have extended their reach beyond campus boundaries through free veterinary intervention campaigns in rural communities, partnerships with municipal authorities, and collaboration with civil society networks, reflecting a broader public health role consistent with One Health principles. Despite their potential, these organizations face structural challenges, including high volunteer turnover, the absence of direct institutional funding, and limited formal integration into governmental frameworks. Institutional support, where present, is predominantly material rather than financial, placing the burden of sustainability on self-management mechanisms. Nonetheless, their embeddedness within academic environments positions them as strategic platforms for interdisciplinary collaboration, experiential training, and community engagement. This study highlights the need to recognize and integrate university-based animal welfare organizations into broader animal population management and public health strategies. Strengthening partnerships between academia, government, and civil society could enhance both animal welfare outcomes and zoonotic disease surveillance efforts in Ecuador and other low- and middle-income countries.

Animals doi: 10.3390/ani16182884

Authors: Zhihua Diao Zhichao Huang Jiangbo Li Jinpeng Cheng Suna Zhao Baohua Zhang

To better analyze the health and welfare of individual dairy cows, this study proposes BR-Tracker, a multi-object tracking method designed for dense monitoring environments to address missed detections, tracking failures, and frequent identity switches. In the object detection stage, a Receptive Field Attention Downsampling (RFADown) module is introduced into the neck network of YOLOv10s to effectively process the locally visible regions of occluded cows by dynamically adjusting the receptive field. An improved Partial Bi-Level Routing Attention (PBRA) module is incorporated into the backbone network to simultaneously extract global and local features, while a Spatial Pyramid Pooling with Efficient Layer Aggregation Network (SPPELAN) module is adopted to enhance multi-scale feature aggregation. In the object tracking stage, an MPDIoU-based matching algorithm is designed to improve matching accuracy and the reliability of trajectory association. The dataset contains 15,420 images for object detection and 130 independent videos for multi-object tracking, of which 40 videos are selected for the final tracking evaluation. Experimental results show that BR-YOLOv10s achieves a precision, recall, and mean average precision (mAP) of 95.7%, 90.3%, and 95.2%, respectively. Compared with the YOLOv10s-ByteTrack baseline, the proposed method improves HOTA, MOTA, MOTP, and IDF1 by 4.4, 5.7, 3.0, and 6.1 percentage points, respectively, while reducing identity switches by 23.19%. In addition, the tracker achieves a processing speed of 43.2 FPS. These results demonstrate that the proposed method can effectively perform real-time detection and tracking of densely distributed dairy cows in complex intensive farming environments.

Animals doi: 10.3390/ani16182883

Authors: Yunfei Wang Yang Zhou Liuxuan Qiao Haoran Zhang Lijing Long Jian Su Chaowei Zhou Zhaofang Han Haiping Liu

Schizothoracinae represent an endemic and dominant teleost group in the Qinghai–Xizang Plateau and adjacent regions. Despite their rich species diversity, closely related Schizothoracinae species exhibit remarkable external morphological similarity, which hinders traditional morphological discrimination. In this study, lapillus otoliths from 360 individuals representing 12 Schizothoracinae species collected across major Tibetan water systems were systematically examined. We evaluated and compared the performance of traditional scalar-based shape indices and Elliptic Fourier Analysis (EFA, 60 normalized harmonic coefficients following size-allometry correction) in otolith contour quantification and species identification. Statistical analyses including one-way ANOVA, principal component analysis (PCA), non-linear manifold learning (t-SNE and UMAP), and linear discriminant analysis (LDA) were executed. One-way ANOVA revealed highly significant differences among all 12 species across all shape indices and Fourier coefficients. The overall LDA classification accuracy was 35.3% for the shape index method, whereas EFA achieved a higher overall accuracy of 68.3%, representing a 33.0 percentage point improvement. Under EFA, Schizothorax lhasaensis (93.3%) and Schizothorax nukiangensis (90.0%) demonstrated the highest single-species discrimination success. Interspecific otolith morphological divergence was substantially higher in the riverine genus Schizothorax than in lacustrine genera (Gymnocypris and Schizopygopsis), the latter two of which show extensive morphological overlap consistent with their low genomic differentiation. Fishes of Gymnocypris displayed significantly higher otolith roundness (p < 0.05), which correlates with their adaptation to lacustrine or sluggish water habitats. Our findings demonstrate that EFA effectively captures multi-scale otolith contour harmonics, serving as a valuable morphometric approach for Schizothoracinae species identification and morphological discrimination, although challenges remain for species with incomplete lineage sorting. The morphometric database established herein provides valuable morphological baselines for the conservation and management of endemic QXP fish resources.

Animals doi: 10.3390/ani16182882

Authors: Xue Yang Xuebing Huang Weiguang Xia Kaichao Li Yanan Zhang Shuang Wang Wei Chen Shenglin Wang Chuntian Zheng

Selenium (Se) is essential in poultry nutrition for antioxidant defense through selenoproteins. While selenomethionine (Se-Met) is commonly used, L-selenomethylselenocysteine (L-SeMC) may offer more favorable responses for selected measured variables. This study compared Se-Met and L-SeMC effects on growth, antioxidant status, and meat quality in broilers. Six hundred and sixty one-day-old male Cobb 500 broiler chickens were randomly assigned to 11 dietary treatments, with six replicate pens of 10 birds per treatment, comprising a basal control diet and 10 selenium-supplemented diets arranged in a 2 × 5 factorial design (Se-Met or L-SeMC at 0.05, 0.10, 0.15, 0.20, or 0.25 mg Se/kg) for 42 days. Selenium supplementation significantly improved average daily gain, final body weight, and feed conversion ratio (p < 0.05). Model-estimated stationary points ranged from 0.137 to 0.142 mg/kg for Se-Met and 0.080–0.126 mg/kg for L-SeMC. Using source-specific model-estimated stationary points, L-SeMC showed 1.85- to 1.93-fold higher biological potency than Se-Met. Birds fed L-SeMC exhibited higher plasma glutathione peroxidase activity, lower malondialdehyde, improved meat color (higher a*_24h), higher ultimate pH, and lower drip loss (p < 0.05). L-SeMC is more effective than Se-Met for enhancing broiler growth, antioxidant capacity, and meat quality.

Animals doi: 10.3390/ani16182881

Authors: Xiaoyu Chen Xiaohong Chu Jing Huang Liang Zhang Fuzeng Lu Nana Yang Lihe Dai Ruhai Xu

The long non-coding RNA XIST is a key regulator of X-chromosome inactivation (XCI) and contributes to epigenetic regulation during cell reprogramming. However, whether XIST suppression influences endogenous pluripotency-associated transcriptional programs in porcine somatic cells remains unclear. In this study, we investigated the effects of XIST knockdown on endogenous pluripotency-associated transcription factors (pTFs) in IBRS-2 cells. Chemically modified antisense oligonucleotides (ASOs) targeting XIST were designed and transfected into IBRS-2 cells. XIST expression was efficiently suppressed for up to 120 h after ASO treatment. RT-qPCR analysis showed that XIST knockdown induced sustained upregulation of KLF4 and LIF expression, prolonged intermittent activation of NANOG, and transient activation of G6PD. In contrast, OCT4, SOX2, and c-MYC were significantly upregulated only during the early stage after transfection. KLF4 immunofluorescence was detectable in a subset of ASO-2-treated cells. These findings demonstrate that ASO-mediated XIST knockdown partially facilitates the activation of a subset of endogenous pluripotency-associated genes in porcine female somatic cells. This study provides new insights into the role of XIST as an epigenetic regulator during porcine somatic reprogramming and suggests that optimized XIST interference may represent a potential strategy for enhancing endogenous pluripotency-associated gene activation without relying on exogenous transcription factor delivery.

Animals doi: 10.3390/ani16182880

Authors: Sara Busechian Dania Cingottini Micaela Sgorbini Simona Orvieto Irene Nocera Francesco Zappulla Fabrizio Rueca

Equine Gastric Ulcer Syndrome (EGUS) and its related diseases, Equine Squamous Gastric Disease (ESGD) and Equine Glandular Gastric Disease (EGGD), can be found in horses worldwide, with increased prevalences in racehorses and animals subjected to heavy training loads. Clinical signs of the disease are vague and nonspecific, and most horses affected by the disease are asymptomatic. In this retrospective study, our aim was to determine the percentages of pleasure and breeding horses showing signs compatible with gastric ulcers, and to correlate these signs with the presence of ESGD, EGGD, and both diseases at the same time. The effects of signalment and activity on disease development and symptoms were also investigated. In total, 1125 adult horses were enrolled in the study. Within this group, only 12% showed clinical signs related to the gastrointestinal tract (GIT, 6%), performance and behavior (PER, 1%), or weight loss (WL, 5%). ESGD was diagnosed in 63% (grade 2/4 22%, 3/4 11%, and 4/4 30%) of animals, while EGGD was diagnosed in 32%. Warmblood horses and geldings were more likely to show clinical signs compared to the other breeds and sexes. WL was associated with the presence and severity of ESGD and the presence of EGGD; GIT was associated only with the presence and severity of ESGD; and PER was associated with the severity of ESGD and the presence of EGGD. Activity was the main risk factor for the development of all diseases, modulating the effect of signalment. EGGD was the only disease with a limited effect on the manifestation of clinical signs when compared with the effect of signalment only. Despite the relatively high prevalence of the disease in horses considered at low risk, clinical signs were present in only a small percentage of the study group, with their expression possibly related to the presence and severity of gastric ulcers. Although the limited effect of EGGD on the manifestation of clinical signs needs to be confirmed, it highlights the limited knowledge we currently have on the clinical significance of lesions in the glandular mucosa, especially when only hyperemia is present.

Animals doi: 10.3390/ani16182879

Authors: Belal S. Obeidat Hadil S. Subih Linda Alyahya

Using industrial waste in livestock feed is crucial for lowering production costs while reducing the environmental impact. Accordingly, the goal of this study was to evaluate the effects of feeding coffee pulp (COP) on the Awassi ewes’ milk production, nutrient digestibility, and blood parameters. Thirty 5-year-old Awassi ewes with an initial body weight of 56.7 ± 3.72 kg were selected for the experiment. Three dietary rations were used, and ten ewes were randomly assigned to each diet for nine weeks, including one week of adaptation. The diets were: (1) 0 COP (CON), (2) 100 COP (COP100), and (3) 200 g/kg COP of dietary dry matter (COP200). Coffee pulp was included at 100 and 200 g/kg of dietary DM; as a result, barley grain increased whereas soybean meal and wheat straw decreased as COP inclusion increased, while the diets remained approximately isonitrogenous. We evaluated effects on intake, performance, milk production and composition, digestibility, nitrogen balance, and selected serum biochemical parameters. Nutrient intake did not differ among dietary groups (p > 0.05). Milk lactose concentration was significantly higher in COP100 and COP200 than in CON (p = 0.015). No adverse changes were detected in the measured productive responses or selected serum biochemical parameters. The cost of milk production decreased with coffee pulp inclusion, from approximately US$ 0.359/kg milk in CON to US$0.325 and US$ 0.300/kg milk in COP100 and COP200, respectively (p = 0.002), corresponding to reductions of approximately 9.5% and 16.4%. Blood urea nitrogen was higher (p = 0.017) in ewes that consumed the COP diets than in those that consumed the CON diet. These results indicated that substituting conventional feedstuffs in ewes’ diets with up to 200 g/kg of COP was effective and reduced milk production costs.

Animals doi: 10.3390/ani16182878

Authors: Fu Tan Liu Zhang Zhuang An Fan

Gangrenous mastitis is a severe form of mastitis in dairy goats that causes extensive tissue damage and has a poor prognosis, thereby substantially affecting the dairy goat industry; however, its molecular pathogenesis remains unclear. In this study, Staphylococcus aureus was used to establish a model of gangrenous mastitis in dairy goats. Mammary gland tissues were collected at 72 h post-inoculation for transcriptomic and proteomic sequencing, followed by integrated multi-omics analyses to systematically identify key regulatory pathways and candidate molecules associated with S. aureus-induced gangrenous mastitis. The results showed that clinical mastitis was characterized mainly by activation of pathways related to the acute inflammatory response, pathogen recognition, neutrophil chemotaxis, and phagocytic defense. In contrast, gangrenous mastitis involved broader molecular reprogramming, with significant enrichment of the TNF, IL-17, and NF-κB signaling pathways, complement and coagulation cascades, platelet activation, and extracellular matrix (ECM) remodeling. Protein–protein interaction (PPI) analysis, gene set enrichment analysis (GSEA), and validation of key molecules indicated that IL6, S100A8, THBS1, SERPINE1, and MMP9 may represent important nodes in disease progression. Collectively, these findings indicate that gangrenous mastitis is a complex infectious tissue-injury process driven by inflammatory amplification, aberrant immune-cell activation, complement–coagulation dysregulation, and tissue structural disruption. The identified molecules and pathways may facilitate the development of biomarker panels for early diagnosis and risk stratification and inform preventive and adjunctive therapeutic strategies targeting excessive inflammation, microcirculatory dysfunction, and ECM damage in dairy goats.

Animals doi: 10.3390/ani16182877

Authors: Margarida Pereira Sofia Alves-Pimenta Carlos Venâncio Maria de Lurdes Pinto Bruno Colaço

Olfactory dysfunction is increasingly recognized as an important consequence of exposure to environmental toxicants, owing to the anatomical and functional characteristics of the olfactory system, which allow inhaled toxicants to reach the central nervous system via the olfactory pathway. Experimental rodent models provide valuable tools for investigating the mechanisms underlying toxicant-induced olfactory injury under controlled conditions. In particular, intranasal vanadium exposure has emerged as a relevant experimental approach because it reproduces an environmentally relevant route of exposure while inducing pathological changes throughout the olfactory system. This review integrates current knowledge derived from experimental rodent models of intranasal vanadium exposure, covering the anatomy and physiology of the olfactory system, intranasal delivery strategies, the cellular and molecular mechanisms underlying vanadium-induced injury, and the associated histopathological, biochemical, behavioural and functional alterations. Vanadium-induced injury is characterized by oxidative stress, mitochondrial dysfunction, neuroinflammation, dysregulation of intracellular signalling pathways, and neuronal degeneration, leading to structural and functional alterations throughout the olfactory pathway. Finally, the review identifies current knowledge gaps regarding olfactory epithelial regeneration and functional recovery following toxic injury and highlights priorities for future research to improve experimental models of environmentally induced olfactory dysfunction.

Animals doi: 10.3390/ani16182873

Authors: Rongyu Yao Fadi Li Xiuxiu Weng Wanhong Li

α-linolenic acid (ALA) has been reported to benefit testicular function. However, dietary ALA is extensively metabolized in the rumen, whereas isolated cell models do not retain seminiferous tubule structure or interactions among testicular cells. This pilot study examined the effects of ALA on seminiferous tubule morphology, UCHL1-positive cells, and gene expression in cultured sheep testicular tissue fragments. Tissue from one 3-month-old Hu ram (n = 1) was cultured for 50 d in control medium (CK) or medium containing 5 μM ALA. Three independently cultured fragments per condition were analyzed. Histology, UCHL1 immunofluorescence, RNA sequencing, and qRT-PCR were performed. ALA-treated fragments showed greater seminiferous tubule diameter, a lower proportion of empty space, and more UCHL1-positive cells per seminiferous tubule. Gene sets related to ATP-dependent chromatin remodeling, DNA replication, and male meiotic nuclear division were enriched in ALA-treated fragments, whereas immune gene sets were enriched in CK fragments. The relative expression levels of SYCP3 and MKI67 were higher in ALA-treated fragments than in CK fragments. Overall, these preliminary findings suggest that ALA treatment was associated with morphological and transcriptional changes in cultured sheep testicular tissue. Further studies using tissues from multiple rams are needed to confirm these findings.

Animals doi: 10.3390/ani16182876

Authors: Luis Barros Sofia Anastácio Ricardo Cabeças Carolina Abrantes Gilberto Fernandes Andreia Gil Catarina Coelho Madalena Vieira-Pinto

Coxiella burnetii, the etiological agent of Q fever, is a globally distributed zoonotic pathogen and a One Health priority. Despite increasing recognition of wildlife as potential reservoirs, data on C. burnetii circulation in European wild ungulates remain scarce. This study estimated exposure to C. burnetii in free-ranging wild boar (Sus scrofa) and red deer (Cervus elaphus) from two major hunting regions in east-central and northeastern Portugal and evaluated the performance of a commercial multi-species ELISA in these hosts. Between 2011 and 2021, 279 serum samples (162 wild boar; 117 red deer) from legally hunted animals in Portugal were tested using an indirect multi-species ELISA using an empirical species-specific threshold derived from a Gaussian mixture model. Overall seroprevalence was 6.8% (CI 95%; 3.4–11.8) in wild boar and 13.7% (CI 95%; 8.0–21.3) in red deer, with higher exposure in wild boar from Beira Baixa (east-central Portugal) than from Trás-os-Montes (northeastern Portugal). The manufacturer’s ruminant cut-off classified almost no animals as seropositive (0.6% and 2.6%, respectively) and showed slight to fair agreement with the species-specific thresholds, underscoring the need for validated, wildlife-appropriate diagnostic criteria. These findings confirm exposure to C. burnetii in both species, suggesting their possible involvement in local maintenance of the pathogen at the wildlife–livestock–human interface. The results reinforce the value of integrating wildlife surveillance into national Q fever surveillance and control strategies to safeguard both public health and livestock production within a One Health context.

Animals doi: 10.3390/ani16182874

Authors: Wanli Li Chen Zhang Pinhui Wu Kangfei Feng Guozhi Zhang Lin Yuan Wei Jin Bingxun Wang Shengli Li Wei Liu Wenqing Li

Indoleamine 2,3-dioxygenase 2 (IDO2) and tryptophan 2,3-dioxygenase (TDO2) are rate-limiting enzymes of the kynurenine pathway, well-characterized in mammals. Unlike all other vertebrates, chickens lack the IDO1 gene and exclusively rely on IDO2/TDO2 for tryptophan (Trp) catabolism; however, their biochemical features and physiological regulatory functions remain poorly defined. Here, we integrated dietary Trp intervention, multi-omics profiling, recombinant enzyme kinetics, and in vitro/inflammatory animal models to characterize chicken IDO2 and TDO2. Supplementary dietary Trp maintained circulating Trp homeostasis while activating hepatic kynurenine metabolism being associated with reduced PPAR signaling and altered lipid and energy balance. Hepatic combined IDO2/TDO2 catalytic activity increased markedly with Trp supplementation; this increase was accompanied by only modest transcriptional upregulation of kynurenine pathway genes (including IDO2, TDO2, and KYNU) and no major changes in their protein abundance, indicating post-translational functional modulation. Recombinant enzyme assays demonstrated optimal activity at chicken physiological pH of 7.2 and temperature of 42 °C; chicken IDO2 displayed measurable catalytic efficiency, which was lower than that of chicken TDO2, whereas its human ortholog showed no detectable activity under the same conditions. Furthermore, LPS and APEC infection significantly suppressed the combined total IDO2/TDO2 activity during inflammatory stress, revealing an avian-specific metabolic–immune regulatory pattern. This work defines the unique biochemical and physiological functions of avian IDO2 and TDO2, offering mechanistic references for poultry nutritional regulation and disease control.

Animals doi: 10.3390/ani16182875

Authors: Xiaoyang Wu Xibao Wang Yongquan Shang Yao Chen Ying Li Ying Gao Guanglin Zhi Weilai Sha Honghai Zhang

Asian black bears (Ursus thibetanus) and brown bears (Ursus arctos) are commonly kept in captivity in zoos, where captive conditions may alter fecal pathogenic bacterial communities and affect host health, although systematic studies remain limited. This study used 16S rRNA sequencing to characterize the composition, diversity, and community assembly mechanisms of fecal pathogenic bacteria in captive brown bears and black bears. Brown bear samples clustered relatively closely, whereas black bear samples were more dispersed, with juveniles and adults clearly separated; the Shannon index of adult black bears was significantly higher than that of brown bears (p < 0.001). Animal pathogenic bacteria (3.37–80.30%) and zoonotic potentially pathogenic bacterial taxa (18.00–96.60%) were the dominant types in both species, with Escherichia coli showing the highest abundance in most samples, and zoonotic potentially pathogenic taxa abundance was higher in adults (83.67%) than in juveniles (74.77%). These findings indicate that captivity and host age shape fecal MBPD-annotated potentially pathogenic bacterial communities in ursids, with enrichment of MBPD-annotated zoonotic potentially pathogenic taxa, including E. coli–assigned sequences, providing a scientific basis for health management of captive bears and potential public health risk assessment.

Animals doi: 10.3390/ani16182872

Authors: Liqin Zhao Mingzhu Wang Yuan Feng Tao Li Cangning Zhang Liang Qu Dan Shao Chengmin Li Manman Shen Weiguo Zhao

In broiler farming, high-fat (HF) diets are frequently incorporated, often leading to hepatic steatosis, oxidative stress, and gut damage. 1-Deoxynojirimycin (DNJ) has emerged as a promising feed additive in animal husbandry due to its significant role in enhancing animal health. This study assessed the protective effects of DNJ-enriched mulberry extract (DNJ) on liver and gut health in broilers subjected to an HF diet, with particular emphasis on the liver-gut interaction. A total of 360 one-day-old Arbor Acres broilers were initially raised on a basal diet for 20 days, after which they were assigned to one of three dietary treatments from days 21 to 42: a control diet (CON), an HF diet, and an HF diet supplemented with 80 mg/kg DNJ (HF + DNJ). By day 42, broilers on the HF diet exhibited increased body weight, feed intake, and body weight gain, alongside reduced feed conversion efficiency. In contrast, DNJ supplementation reduced body weight without affecting feed intake in the HF diet group. It lowered liver T-CHO and LDL-C levels, increased SOD and CAT activities, and decreased MDA. Histological analyses showed that DNJ preserved intestinal mucosal structure and epithelial tight junctions. DNJ supplementation enriched microbes like Actinobacteriota and Baceroidota and reversed the HF-induced Firmicutes-to-Bacteroidota ratio. Furthermore, Bifidobacterium pullorum and Bacteroidota were positively correlated with antioxidant markers. In conclusion, dietary DNJ-enriched mulberry extract serves as an effective functional feed substance that mitigates HF-induced metabolic syndromes in broilers by balancing lipid metabolism, enhancing antioxidant defenses, safeguarding intestinal barrier integrity, and restoring healthy microbial community structures along the liver-gut axis.

Animals doi: 10.3390/ani16182871

Authors: Zeynep Yerlikaya Burak Karabulut Eren Cankaya Muhammed Girgin Pelin Fatos Polat Dincer Hatice Eroksuz

Ovine pneumonia is a multifactorial respiratory disease, yet bacterial community alterations associated with pathologically confirmed pneumonia in lambs remain poorly characterised. This study investigated the lung bacterial microbiota of non-pneumonic and pneumonic Akkaraman lambs by integrating gross and histopathological examination with 16S rRNA gene amplicon sequencing. The final dataset comprised lung samples from 15 non-pneumonic and 14 pneumonic lambs. Alpha diversity did not differ significantly between groups, whereas bacterial community composition differed significantly for both Bray–Curtis (R2 = 0.093, p = 0.001) and weighted UniFrac (R2 = 0.117, p = 0.006) distances, with no significant differences in multivariate dispersion. At the genus level, Mannheimia showed significantly higher relative abundance in pneumonic lungs after false-discovery-rate correction and was the only genus also identified as differentially abundant by ANCOM. Mycoplasma, Fusobacterium, and Trueperella showed greater relative representation in pneumonic lungs, whereas Acinetobacter and Psychrobacter showed the opposite pattern. Prevalence analysis further demonstrated contrasting taxon distributions between groups. Overall, naturally occurring pneumonia was associated with distinct alterations in lung bacterial community composition rather than a generalised loss of within-sample diversity, with Mannheimia representing the strongest pneumonia-associated genus-level signal.

Animals doi: 10.3390/ani16182870

Authors: Ziluo Chen Yaya Zhao Mingwei Bao Fangyi Zhou Qingzhong Shen Xianming Guo Li Zhang

Asian elephants (Elephas maximus) face substantial thermoregulatory constraints because of their large body size, low relative surface area, sparse hair, and lack of functional sweat glands. Reliable body temperature measurement is essential for assessing thermal status and evaluating welfare in both wild and managed populations, but conventional rectal thermometry requires close physical contact, animal training, and repeated manual handling, making high-frequency, continuous, large-scale monitoring impractical. This study developed a non-invasive framework for automatically detecting the outer ear and extracting auricular surface temperature from infrared thermograms. Rectal temperature, regional surface temperatures, ambient temperature, and relative humidity were measured synchronously in eight semi-captive Asian elephants, yielding 425 matched observations. The associations between rectal temperature and the surface temperatures of three anatomical regions (head, outer ear, torso and limbs) were analyzed using repeated-measures correlation accounting for the non-independence of repeated measurements. Mean outer-ear temperature showed the strongest within-individual association with rectal temperature (rrm = 0.395, p < 0.001), identifying the outer ear as the optimal thermal window for subsequent automated monitoring. Eight lightweight YOLO models—YOLOv5n, YOLOv5s, YOLOv8n, YOLOv8s, YOLO11n, YOLO11s, YOLO26n, and YOLO26s—were trained on 2178 annotated infrared images and evaluated on an independent 194-image test set from extra elephants. Model performance was assessed using detection metrics, inference speed, Bland–Altman agreement, Taylor diagram statistics, and a weighted multi-criteria score with Monte Carlo sensitivity analysis. YOLO11n achieved the best overall performance, with an mAP50 of 0.933 and an inference speed of 164 frames per second. The proposed framework provides an efficient method for automated auricular temperature monitoring and has potential applications in elephant welfare management and remote physiological surveillance.

Animals doi: 10.3390/ani16182868

Authors: Liping Gan Yifeng Zhao Shiya Li Yingying Sheng Dongtian Zhao Yuan Yuan Hanzhen Qiao Peng Wang Erzhen Duan Tengpeng Liu Jinrong Wang

Enterotoxigenic Escherichia coli (ETEC) infection severely impairs post-weaning piglet health, causing intestinal dysfunction, systemic inflammation, and significant economic losses in the swine industry. This study aims to evaluate a synthesized palmatine-intercalated montmorillonite composite (MP) for improving growth performance, immune response, and intestinal health in ETEC-challenged piglets. Following structural characterization, an in vivo trial was conducted using 30 crossbred piglets (Yorkshire × Landrace × Duroc; 15.12 ± 2.95 kg BW) challenged with ETEC K88 and supplemented with 0.02%, 0.5%, and 1.5% MP. Structural analysis confirmed successful intercalation of palmatine into montmorillonite, with enhanced antibacterial and anti-inflammatory properties. In vivo, dietary inclusion of the 1.5% MP significantly alleviated ETEC-induced growth depression, as evidenced by increased average daily gain and improved apparent total tract digestibility of crude protein. These effects were associated with improved jejunal morphology and upregulation of PEPT1, B0AT1, and SGLT1. Moreover, MP supplementation attenuated systemic inflammation and hepatic stress, as indicated by reduced serum AST, TNF-α, and IL-1β levels. Microbiota analysis showed that MP modulated ETEC-associated alterations in the cecal microbial community. In conclusion, the MP represents a promising nutritional approach to improve intestinal resilience, nutrient utilization, and growth performance of ETEC-challenged piglets.

Animals doi: 10.3390/ani16182869

Authors: Hao Dong Xiaofan Fang Yuefan Zhang Shubo Jin Wenyi Zhang Yiwei Xiong Hui Qiao Sufei Jiang

This study evaluated the genetic diversity and population structure of wild M. nipponense populations in the Pearl River Basin based on mitochondrial DNA (mtDNA) D-loop sequences. A total of 14 wild populations were collected from the West River, North River, and Pearl River Delta, and genetic diversity and population differentiation were assessed using haplotype analysis, analysis of molecular variance (AMOVA), pairwise FST, and principal component analysis (PCA). A total of 351 variable sites were identified. The overall haplotype diversity (h) and nucleotide diversity (π) were 0.947 and 0.03665, respectively, indicating high genetic diversity across the Pearl River Basin. Significant genetic differentiation was detected among populations (FST = 0.3785, p-value < 0.001), with 37.85% of genetic variation occurring among populations. PCA revealed distinct genetic relationships among populations, supporting the presence of population genetic structure within the Pearl River Basin. Populations from Tian’e County, Xilin County, Dahua Yao Autonomous County, and other upper tributaries of the West River exhibited distinct genetic characteristics compared with populations from Zhaoqing City, Guiping City, Guangzhou City, and the middle and lower reaches of the West River and the Pearl River estuary. Among the 14 populations, those from Zhaoqing City, Guiping City, and Guangzhou City exhibited relatively high genetic diversity and distinctive genetic characteristics. These findings suggest that the complex hydrological structure and historical connectivity of the Pearl River Basin have contributed to the observed population genetic structure. The genetically diverse populations identified in this study represent valuable wild germplasm resources for the conservation and selective breeding of M. nipponense.

Animals doi: 10.3390/ani16182867

Authors: Paul D. McGreevy David J. Mellor Dan O’Neill Rowena M. Packer Cathrynne Henshall Cristina L. Wilkins

The domestic dog is the most morphologically diverse domesticated mammalian species. That diversity has arisen from deliberate, sustained artificial selection over the most recent 1% of the domestic dog’s evolutionary history. However, artificial selection has pushed many individual breeds so far from the ancestral canine form that many dogs within these breeds can no longer perform the critical biological functions their bodies evolved to serve. Indeed, much of the popularity of many dog breeds is predicated on them no longer being functional dogs; rather, they are representations or caricatures of human aesthetic desires. The result is breeds in which the teleonome of individual dogs, i.e., the integrated biological system by which they maintain viability has been structurally dismantled in the pursuit of appearance. This article applies teleonomic and allostatic load theory to a multi-domain analysis of extreme brachycephaly as a primary case study, drawing on representative examples from other extreme phenotypes to examine their welfare costs across conformational disorder, innate health, allostatic load, and veterinary costs. A detailed Five Domains analysis of extreme brachycephalic morphotypes, where the epidemiological evidence is most complete, reveals that every major welfare domain is simultaneously compromised from birth because artificial selection has structurally compromised teleonomic function across morphological, physiological, physical, and behavioural traits. Allostatic load theory then provides the mechanistic explanation for the resulting shorter lifespans. The article concludes by addressing the veterinarian’s emerging role as arbiter of animals’ fitness for purpose under consumer and welfare laws, making the case for an Innate Health Assessment to become a standardised functional assessment framework grounded in teleonomic principles.

Animals doi: 10.3390/ani16182866

Authors: Saleem Ahmad Jinyoung Park Jin-ho Jeong Seung-Bum Kang Kyung-Duk Min Dae Sung Yoo

Lumpy skin disease (LSD) is a rapidly expanding transboundary vector-borne disease threatening livestock systems across Asia, with increasing risk of introduction into previously unaffected regions such as South Korea. This study developed an integrated, spatially explicit framework to assess the plausibility, timing and source-region structure of LSD virus (LSDV) introduction via windborne dispersal of infected arthropod vectors. Maximum Entropy models based on climatic, environmental, and topographic variables were used to determine habitat suitability for four major vector families (Culex spp., Musca domestica, Stomoxys calcitrans, and Culicoides spp.). All four were run via the transport simulation; S. calcitrans had the best experimental transmission efficiency, whereas Culicoides spp. provides the strongest evidence for passive long-distance transport. Livestock density and vector suitability were used to calculate the risk of LSD incidence in the adjacent source regions. A fourth-order Runge–Kutta trajectory model driven by ERA5 reanalysis winds at 850 hPa (about 1458 m) was used to predict windborne dissemination. This model included degradation of mechanically transmitted virus as well as survival limitations based on temperature, humidity, and wind. The simulated trajectories arrived in South Korea within the retention period of mechanically transported LSDV, with a median transit time of 18 h (interquartile range 10.0–30.3 h), 61.4% arriving within 24 h, and 87.1% within 48 h. North Korea accounted for 76.5% of weighted arrivals, with a median transit time of 11 h. Transit durations varied significantly by source location. For all four vector species, the predicted infectious arrival mass peaked in September and decreased thirteen-fold by November. Infected farms were found in regions of greater anticipated exposure than non-infected controls when compared to the October 2023 Korean epidemic (58.1% vs. 34.3% in the two highest risk groups; median exposure 0.214 versus 0.045; p < 0.001), although randomly relocating the dispersal surface reproduced comparable agreement in 23.6% of permutations, indicating that outbreak locations provide only limited validation of a pathway-specific introduction model. Random permutation of the dispersion surface produced a comparable result in 23.6% of permutations, indicating limited robustness of the observed spatial relationship to spatial randomization. With pairwise geographical and temporal distances significantly associated (Mantel r = +0.136, p = 0.013), spatiotemporal analysis revealed that the 74 reported outbreaks originated from a maximum of 25 introduction events over a 13-day period, consistent with significant farm-to-farm dissemination after introduction. The seasonal window, source-region structure, and transit timeframes of windborne LSDV introduction into South Korea are described in this paper. Reported epidemic sites only partially validate a pathway-specific introduction model since they reflect both the point of introduction and subsequent local transmission. This evaluation is pathway-specific and does not evaluate overall incursion risk; windborne transport is one of numerous possible introduction paths to consider.

Animals doi: 10.3390/ani16182865

Authors: Mahmoud M. Shaaban Min Gao Mohamed El-Sherbiny Ahmed M. Abd El Tawab Mostafa S. A. Khattab Mohamed A. Radwan Fatma I. Hadhoud Yongbin Liu

This study evaluated a rumen-derived Streptococcus macedonicus isolate in Barki lambs using in vitro and in vivo approaches. The isolate was characterized for antibacterial activity and antimicrobial susceptibility. In vitro, a control diet was incubated for 24 h with 0, 0.002, 0.004, 0.008, or 0.016 g of S. macedonicus, and fermentation and digestibility were determined. In vivo, 40 Barki lambs were assigned to four dietary treatments for 210 d: control, control plus 1 g/d commercial probiotic, or control plus 1 or 2 g/d S. macedonicus. Feed intake, nutrient digestibility, growth performance, carcass traits, and meat quality were evaluated. The isolate showed antibacterial activity against Escherichia coli, Pseudomonas aeruginosa, and Bacillus cereus. In vitro, S. macedonicus increased dry matter digestibility (52.9% to 56.7%; p = 0.042) and total gas production (p < 0.0001). In vivo, dry matter digestibility increased from 66.3% to 75.2% (p = 0.002). Supplemented lambs had greater final body weight, average daily gain, and an improved feed conversion ratio (p < 0.001). Supplementation also improved carcass yield, dressing percentage, longissimus muscle area, lean proportion, water-holding capacity, and meat redness while reducing carcass fat percentage. Overall, rumen-derived S. macedonicus improved nutrient utilization, growth performance, and carcass and meat quality traits.

Animals doi: 10.3390/ani16182864

Authors: Maxat Toishimanov Gulmira Bekova Dastanbek Baimukanov Dinara Begaliyeva Anastasiya Yanina Rakhim Kanat Dias Daurov Ainash Daurova Zagipa Sapakhova Gemingguli Muhatai Zhaniya Assanbek Alzhan Shamshidin Indira Beishova Malika Shamekova

Mare’s milk is recognized for its nutritional value, but information on the relationships of mare age and number of foalings with body conformation, udder morphology, milk yield, and milk composition in the indigenous Kushum breed remains limited. Forty-one clinically healthy lactating Kushum mares were evaluated in June and classified by age (4–7, 8–11, and 12–16 years) and number of foalings (1–3, 4–7, and 8–11). Body traits were measured once; udder morphometry was available for 30 mares. Milk yield was recorded at 08:00, 10:00, 12:00, 14:00, and 16:00 h on two consecutive days. On Day 2, a 100-mL aliquot was collected after each milking, and the five aliquots were pooled into one 500-mL composite sample per mare; physicochemical composition was available for 40 mares. Age and number-of-foalings effects were evaluated in separate models. Continuous morphometric and physicochemical traits were analyzed by one-way ANOVA with Tukey-adjusted comparisons, with Benjamini–Hochberg false discovery rate (FDR) correction across trait families; two-day milk yield was additionally evaluated with a repeated-measures mixed model. After FDR correction, cannon bone circumference, body weight, udder length, and udder width differed among both age and number-of-foalings groups. Teat shape also differed among groups. The two-day mean daily milk yield did not differ by age (p = 0.483) or number of foalings (p = 0.195), although significant group-by-day interactions were detected. In pooled Day-2 milk, SNF, lactose, glucose, freezing point, acidity, lactic acid, density, and urea differed among both classification factors after FDR correction (q < 0.05), whereas fat, protein, total solids, galactose, citric acid, and casein did not. PCA explained 60.9% of the variance in body/udder traits plus mean milk yield and 68.8% of the variance in physicochemical composition. These findings indicate selective associations of age and number of foalings with Kushum mare phenotype and milk composition, while emphasizing the limitations of small udder subgroups and equal-volume pooled sampling.

Animals doi: 10.3390/ani16182863

Authors: Slawomir Gonkowski Manolis Tzatzarakis Elena Vakonaki Liliana Rytel

Triclosan (TCS) and triclocarban (TCC) are antimicrobial compounds widely used in numerous industries, making them significant environmental pollutants that infiltrate living organisms and exhibit endocrine-disrupting activity. However, knowledge regarding the exposure of wild terrestrial mammals to these substances remains virtually non-existent. Therefore, this study analyzed, for the first time, TCS and TCC levels in the hair of wild boars using liquid chromatography-tandem mass spectrometry (LC-MS). Despite widespread environmental contamination with these compounds, concentrations above the limit of detection (LOD: 2.3 pg/mg for TCS and 0.8 pg/mg for TCC) were detected in only a small percentage of the analyzed samples: 14.81% for TCC and 3.70% for TCS. Higher concentrations and a greater frequency of TCC occurrence were observed in animals from highly urbanized and industrialized regions. In contrast, TCC and TCS concentrations in animals from other areas only sporadically exceeded the LOD. These results suggest that wild boars are primarily exposed to TCC in highly urbanized areas. In less-urbanized regions, animal exposure to both compounds is usually below the LOD and likely depends on poorly defined local factors. Nonetheless, fully clarifying the exposure of wild terrestrial mammals to TCC and TCS requires further comprehensive research.

Animals doi: 10.3390/ani16182862

Authors: Guang Yi Xilin Wang Songyu Jiang Jianfeng Zhao Tengfei He Zhaohui Chen

Continuous multi-parameter monitoring is important for characterizing environmental conditions in commercial beef cattle facilities, but sensor performance and system reliability require evaluation under production conditions. This study developed a LoRa-based multi-sensor node for air temperature, relative humidity, CO2, NH3, air speed, and illuminance. Field performance was evaluated at one commercial farm by synchronous comparison with commercial instruments using 1-min paired observations and regression- and agreement-based analyses. Nine prototype nodes were subsequently deployed at three sites for 14-day monitoring periods. Temperature and relative humidity showed the strongest linear relationships with the comparison instruments (R2 = 0.995 and 0.994), followed by illuminance, air speed, and CO2 (R2 = 0.990, 0.863, and 0.773). NH3 showed a weaker relationship (R2 = 0.604), improving after 10-min aggregation; most application-monitoring estimates below 5 ppm were extrapolated rather than validated absolute concentrations. Overall data availability was 99.66%, and continuous monitoring captured temporal patterns in thermal conditions, air quality, local airflow, and illuminance. The evaluation focused on environmental monitoring performance rather than animal-based outcomes. These findings support parameter-specific environmental monitoring and multi-site deployment in commercial beef cattle facilities, with potential for future anomaly detection and environmental control.

Animals doi: 10.3390/ani16182861

Authors: Zhiyuan Xu Xuanlin Sun Yan Liang Xuan Tao Bo Ran Pengliang Liu Yiren Gu

Tibetan pigs exhibit remarkable adaptability to hypoxic high-altitude conditions. However, metatranscriptomics has not been used to investigate the transcriptional activity and functional responses of the gastrointestinal microbiota as part of their adaptation to high altitude. We collected 57 samples of the contents of the stomach, jejunum, cecum, colon, and rectum of six high-altitude Tibetan pigs (n = 6, elevation 3750 m) and six low-altitude black pigs (n = 6, elevation 500 m) for metatranscriptomic analysis. We identified 79 differentially abundant bacterial genera and 69 differentially abundant bacterial species. In particular, Lactobacillus crispatus and Lactobacillus delbrueckii were significantly more abundant in the gastrointestinal microbiota of the high-altitude Tibetan pigs than in that of the low-altitude black pigs. Functional analysis revealed differences in transcripts associated with energy, carbohydrate, and amino acid metabolism. The microbial correlation network of Tibetan pigs contained more nodes and edges than that of black pigs. Consistent with this, within the CAZyme database, the CBM32 family of carbohydrate-binding modules and GH27 glycosidases were significantly enriched in the Tibetan pig samples, suggesting that these enzymes may play a crucial role in energy acquisition and carbon source metabolism in these pigs. These findings provide insights into the gastrointestinal microbial characteristics of Tibetan pigs and black pigs. In summary, these changes may enhance the stability of the gut ecosystem of Tibetan pigs and improve their energy utilization in nutrient-deficient high-altitude environments. These findings suggest that the gut microbiota may contribute to metabolic regulation in Tibetan pigs at high altitude and provide important clues regarding the gastrointestinal adaptations of high-altitude animals.

Animals doi: 10.3390/ani16182860

Authors: Alberto Prieto Silvia Valverde Pablo Díaz José Manuel Díaz-Cao Gonzalo López-Lorenzo David García-Dios Renato Barrionuevo Gonzalo Fernández

Canine parvovirosis is a fatal disease state traditionally caused by canine parvovirus (CPV), which evolved from feline panleukopenia virus (FPV). While FPV was historically restricted to felids, FPV-like strains were recently reported to be causing clinical disease in dogs. This study aims to characterize an FPV-like strain detected in an eight-week-old unvaccinated female dog in Spain exhibiting hemorrhagic diarrhea and severe leukopenia. After fecal DNA isolation, a 1745-base-pair fragment of the VP2 gene was sequenced for phylogenetic and molecular analysis. Our results revealed that this strain shared high homology with local feline FPV strains; the sequence was clustered near to other FPV strains detected in dogs elsewhere. Molecular analysis identified an isoleucine-to-threonine substitution at position 101 (I101T) of the VP2 protein. This mutation, common in contemporary FPV and CPV variants, has been described as a structural modification that optimizes interaction with the canine transferrin receptor, suggesting a potential role in canine host adaptation. In conclusion, this study represents the first report in Spain of FPV associated with clinical gastroenteritis in dogs. These findings underscore the necessity of continuous genomic surveillance of these highly evolving viruses for monitoring host-switching events.

Animals doi: 10.3390/ani16182859

Authors: Yaxin Di Xiaona Jiang Chitao Li Xuesong Hu Yanlong Ge Xiaodan Shi Xinyu Zhao Zhiying Jia

Tumor necrosis factor receptor-associated factors (TRAFs) are key signal transduction molecules in innate immunity and are involved in multiple immune pathways, such as TNF receptor and Toll-like receptor signaling. In this study, 16 CcTRAFs were identified from the common carp genome, assigned to six subfamilies (TRAF1/2/3/4/6/7), with no TRAF5 found. Significant paralogous expansion was observed in the TRAF2/4 subfamilies. All CcTRAF proteins were predicted to be unstable and hydrophilic by ProtParam analysis; most contain conserved RING, TRAF and MATH domains, whereas CcTRAF7 carries seven WD40 domains. Gene structure and conserved motif analysis revealed high conservation of exon numbers and motif compositions within each subfamily, while the TRAF7 subfamily displayed distinct characteristics. Collinearity analysis further indicated that segmental duplication promotes CcTRAF family expansion. qRT-PCR showed that CcTRAF genes have tissue-specific expression patterns and are preferentially expressed in the liver, spleen, and kidney. Under CyHV-3 infection, the expression levels of CcTRAF3/6/7 in these tissues first increased and then decreased, peaking at 48 h post-challenge. In summary, CcTRAF genes, especially CcTRAF3/6/7, play important roles in the immune response of common carp against CyHV-3. This study provides a molecular basis for understanding innate immune regulation in teleost fish and the functional roles of TRAF genes.

Animals doi: 10.3390/ani16182858

Authors: Xiaotong Luo Hongliang Liu Xuanyu Li Chunyu Li Congcong Zhang Dong Li Yongsheng Yu Jian Wu

Intramuscular fat (IMF) deposition drives premium beef value, but phenotypic selection is costly and slow. Although genome-wide studies have linked the KIRREL3 region to meat quality, whether intronic variants directly affect carcass traits and retain predictive power across physiological stages remains unproven. We genotyped two intronic SNPs (g.29756010C>T and g.29756149C>T) in Woking cattle, a high-quality beef breed developed in Jilin Province, China (n = 473; 226 and 206 heifers at 36 and 48 months, respectively, plus an independent validation cohort of n = 41), assessed for fatty acid composition, meat quality, and multi-tissue gene expression. Both loci were polymorphic and in Hardy–Weinberg equilibrium. At 36 months, TT individuals showed higher dressing percentage, rib thickness, and meat grade than CC at both loci. By 48 months, the 63-bp locus retained broader associations with carcass traits, whereas the 203-bp locus effects weakened. In the validation group, CC animals had higher IMF and muscle KIRREL3 expression than TT, yet received lower meat grades. Five fatty acids differed significantly among genotypes after Benjamini–Hochberg correction (q < 0.05), whereas the dominant fatty acids (C16:0, C18:1n9c, C18:0, C18:2n6c) showed no proportional differences. KIRREL3 was most highly expressed in the kidney among eight tissues examined. These findings identify the 63-bp site as a candidate marker for meat quality in local Chinese cattle, with potential for marker-assisted selection pending confirmation in larger cohorts and functional studies.

Animals doi: 10.3390/ani16182857

Authors: Batya G. Jaffe

Animal-Assisted Crisis Response (AACR) is a specialized form of crisis intervention in which trained handlers and therapy dogs provide emotional support following crises and disasters. Although previous research has examined AACR, less is known about the contribution of the human–canine partnership. This study explored the human–canine partnership and its perceived role in effective crisis response and canine welfare. This qualitative study used semi-structured interviews and demographic surveys. Twenty-one crisis responders participated, and interview data were analyzed using Charmaz’s grounded theory approach. Findings suggest that participants perceived a highly trained handler, a carefully selected dog, and the quality of their partnership as important components of effective AACR practice. Canine welfare emerged as a fundamental component of the human–canine partnership, with participants emphasizing the handler’s role in recognizing canine stress cues, safeguarding welfare, and cultivating a strong partnership with the dog. Although bringing a dog to a crisis scene presents practical challenges, participants also described benefits for the team and improved engagement with trauma survivors. These findings support conceptualizing AACR as a welfare-centered human–canine partnership in which handler expertise, canine suitability, and attention to welfare are perceived as important components of effective and ethical crisis-response practice.

Animals doi: 10.3390/ani16182856

Authors: Anna Michalska Jakub Kaczmarek Daniel Kaup Annika Lehmbecker Magdalena Morawska

Aneurysmal bone cysts (ABCs) are benign but locally aggressive osteolytic lesions that are rarely reported in dogs and may progress or recur following surgical treatment. This case report describes the diagnosis, surgical management, postoperative progression, and long-term outcome of an ABC affecting the left ulna of a four-month-old Labrador Retriever. Radiography and computed tomography revealed a rapidly progressive, expansile osteolytic lesion with marked cortical thinning. Histopathological examination supported the diagnosis of ABC, with no evidence of malignancy. Initial treatment consisted of corticotomy, intralesional curettage, autologous bone grafting, and stabilization of the cortical segment with lag screws. Despite complete clinical resolution of lameness, surveillance radiographs six weeks postoperatively demonstrated progression of the lesion proximal to the original surgical site. Revision surgery involving a larger cortical window and more extensive curettage was subsequently performed, followed by bone grafting. Histopathology again confirmed ABC without malignant transformation. The dog regained normal limb function, and serial radiographic and computed tomographic examinations demonstrated progressive osseous remodeling with no evidence of further progression or recurrence through 35 months of follow-up. This case highlights the importance of adequate surgical excision and scheduled postoperative imaging, as ABC progression may occur despite apparent clinical recovery.

Animals doi: 10.3390/ani16182855

Authors: Min Jae Kim Jae-Ok Kim Hee-Jung Choi Mun-Gyeong Kwon Chan-Il Park Do-Hyung Kim Kwang-Il Kim

In pond-based shrimp culture, seawater exchange may introduce white spot syndrome virus (WSSV) from coastal waters, but infection risk at a given seawater viral concentration may vary with temperature, stocking density, and exposure duration. This study evaluated waterborne WSSV susceptibility in whiteleg shrimp (Penaeus vannamei) under these conditions. Shrimp were reared at 25 or 30 °C and at low (70), medium (140), or high (210 shrimp/m2) stocking densities; physiological responses were assessed, followed by short-term and continuous immersion challenges and a cohabitation challenge. Higher stocking-density conditions were associated with stress-related physiological changes and greater WSSV susceptibility. Under short-term exposure, 107 WSSV genome copies/L caused infection only in the high-density group at 25 °C. Under continuous exposure, the minimum infective dose (MID) values were 105 and 104 WSSV genome copies/L at low and high stocking densities at 25 °C, and 106 and 105 WSSV genome copies/L, respectively, at 30 °C. Cohabitation transmission occurred at seawater viral loads below those associated with short-term immersion infection. By linking seawater viral concentrations with experimentally defined MID values and cohabitation-transmission outcomes, these results indicate that waterborne WSSV transmission risk should be assessed in relation to viral concentration, exposure duration, water temperature, and stocking-density conditions.

Animals doi: 10.3390/ani16182854

Authors: Zhending Gan Jiawei He Qiyue Jin Qiuqin Ma Chuanlong Wang Xiang Zhong

Acute inflammatory challenge can impair intestinal barrier function and bile acid homeostasis in weaned piglets. Glutamate serves as an important energy substrate and functional amino acid for intestinal epithelial cells, yet the epitranscriptomic mechanisms underlying its potential protective effects remain poorly defined. This study investigated whether dietary glutamate supplementation is associated with m6A-dependent regulation of SLC10A2 and bile acid signaling in weaned piglets subjected to an acute LPS challenge, with supporting validation in IPEC-J2 cells. LPS challenge (100 μg/kg body weight) impaired ileal morphological structure, activated the TLR4/NF-κB signaling pathway, and aggravated intestinal inflammatory responses in piglets. In contrast, glutamate supplementation was associated with restored ileal mucosal morphology, up-regulated expression of intestinal tight junction proteins, and suppressed TLR4/NF-κB pathway activation. Further metabolomic analysis indicated that LPS stimulation and glutamate intervention altered ileal bile acid metabolism: LPS decreased total bile acid (TBA) content in ileal tissue while increasing TBA accumulation in ileal chyme, whereas glutamate treatment partially reversed this pattern, by elevating ileal tissue TBA and reducing chyme TBA, suggesting a facilitatory effect of glutamate on intestinal bile acid reabsorption. Glutamate supplementation was also associated with increased ileal ASBT (SLC10A2) and FXR (NR1H4) expression. The SLC10A2 transcript exhibited detectable m6A modification; glutamate supplementation rescued the LPS-induced downregulation of m6A demethylases FTO and ALKBH5, which was associated with reduced global and SLC10A2-specific m6A modification. In vitro experiments in IPEC-J2 cells suggested that inhibition of FTO or glutamate dehydrogenase attenuated the effects of glutamate on m6A modification and SLC10A2 expression, and that YTHDF2 may contribute to the degradation of hyper-m6A-modified SLC10A2 mRNA. Glutamate-derived α-KG may serve as a cofactor supporting demethylase expression and activity after LPS stimulation. Collectively, these findings suggest that glutamate may alleviate LPS-induced intestinal inflammation in weaned piglets through a pathway involving α-KG-dependent maintenance of m6A demethylase expression, reduced m6A modification of SLC10A2, and preserved ASBT/FXR signaling. This study provides evidence for a nutritional–epitranscriptomic axis that may contribute to intestinal protection under acute inflammatory stress.

Animals doi: 10.3390/ani16182853

Authors: Yu Shi Bingang Shi Youpeng Qi Changze Cui Meixian Zhang Zhihao Luo Yuwei Ma Jiang Hu

The molecular mechanisms underlying testicular development in the yak remain poorly understood. In this study, RNA sequencing was performed on testicular tissues from Tianzhu white yaks at three postnatal developmental stages: 0.5 (prepubertal), 2.5 (pubertal), and 4.5 (adult) years, with six animals in each group. Differential expression analysis was integrated with weighted gene co-expression network analysis (WGCNA) to characterize transcriptomic changes associated with testicular maturation. A total of 9646 differentially expressed genes (DEGs) were identified between the 0.5- and 2.5-year groups, whereas only 126 DEGs were detected between the 2.5- and 4.5-year groups, indicating that the transition to puberty represents the most dynamic phase of transcriptional reprogramming during testicular development. Functional enrichment analysis revealed that focal adhesion and extracellular matrix–receptor interaction were the most significantly enriched Kyoto Encyclopedia of Genes and Genomes pathways, suggesting a potential role for extracellular matrix (ECM) remodeling in the establishment of the blood–testis barrier and testicular maturation. WGCNA identified seven age-associated co-expression modules. The turquoise module showed the strongest positive correlation with developmental stage and was significantly enriched for genes involved in spermatid differentiation and maturation, including SPATA6, SPMIP11, TNP1, and ODF1. Collectively, these findings provide a comprehensive transcriptomic characterization of yak testicular development.

Animals doi: 10.3390/ani16182852

Authors: S. Ferrando-Juan J. Gómez-Aguilera A. Tomás-Vidal M. Rodilla M. Jover-Cerdá J. Calanche S. Martínez-Llorens D. S. Peñaranda

Fishmeal (FM) reduction can improve shrimp feed sustainability, but low FM formulations remain insufficiently validated across production stages, scales, and post-harvest quality traits. This study evaluated alternative protein strategies for Penaeus vannamei reared under super-intensive biofloc technology (BFT) at different production scales. In a medium-scale grow-out trial, a Control diet containing 15% FM was compared with diets containing 7.5% FM using animal-derived proteins (A7.5) or combined animal- and plant-derived proteins (AP7.5). AP7.5 maintained growth and apparent feed conversion comparable to those of the Control diet, whereas A7.5 showed poorer performance. Post-harvest quality was assessed in commercial-size shrimp during refrigerated storage without antimelanosis treatment. Sensory differences were limited shortly after harvest but became more evident during storage, with AP7.5 maintaining a comparatively stable sensory profile and showing lower panel-assessed melanosis after seven days. Subsequently, the AP7.5 diet was validated against the Control in a pilot large-scale trial covering the juvenile and grow-out phases, maintaining comparable growth, economic conversion ratio, and profitability index. Sensitivity analysis further showed that its relative economic performance was maintained across simulated FM and shrimp price scenarios and became more favorable as FM prices increased. In conclusion, the consistent productive and economic performance of AP7.5 across production stages and scales, together with maintained sensory quality and lower panel-assessed melanosis, supports its potential as a scalable low-FM strategy for BFT shrimp production.

Animals doi: 10.3390/ani16182851

Authors: Jun Bai Yiru Wang Xinhang Li Jiajun Li Zhenlong Wu Xinjian Li Kejun Wang Ruimin Qiao Tong Fu Yanpeng Li

During swine gestation, the placenta serves as the sole interface for nutrient, gas, and signaling molecule exchange between the dam and fetuses, and its function critically determines pregnancy outcomes. Branched-chain amino acids (BCAAs)—leucine, isoleucine, and valine—are substrates for protein synthesis, precursors for glutamate and glutathione, and signaling molecules that activate the mechanistic target of rapamycin (mTOR) pathway. This review synthesizes current evidence on BCAA transport, catabolism, and signaling in the porcine placenta, organized around four mechanistic axes: transporter-mediated transplacental transfer, local catabolism supporting energy production and antioxidant defense, mTORC1-dependent regulation of growth and differentiation, and modulation of endocrine and immune homeostasis. Both deficiency and excess of BCAAs compromise placental function, contributing to intrauterine growth restriction and metabolic programming abnormalities through a U-shaped dose–response relationship. Critical knowledge gaps remain, including the reliance on rodent and in vitro models, the absence of standardized supplementation protocols and dose–response data, unaddressed confounders (parity, litter size, environment), and the lack of formal power analyses or meta-analytic synthesis. We propose precision dietary strategies to optimize BCAA supplementation during gestation for improved reproductive performance in swine.

Animals doi: 10.3390/ani16182850

Authors: Ueli Braun Karl Nuss Christian Gerspach

The laboratory and ultrasonographic findings in 1163 cattle with one of 11 types of ileus, including caecal dilatation, compression, haemorrhagic bowel syndrome, mesenteric torsion, paralytic ileus, incarceration, intussusception, obstruction, strangulation, volvulus, and torsion of the spiral colon were compared. The prevalences of the individual laboratory and ultrasonographic findings varied greatly among the different types of ileus; hypochloraemia ranged from 98.4 to 12.4%, hypokalaemia from 98.4 to 42.4%, azotaemia from 91.9 to 38.3%, acidosis from 47.8 to 3.3%, and haemoconcentration from 71.7 to 23.2%. Ultrasonographically, dilated small intestine varied from 100 to 43.3%, intestinal atony from 98.4 to 0%, free abdominal fluid from 66.7 to 16.0%, abomasal dilatation from 44.4 to 0%, and empty poststenotic small intestine from 34.9 to 0%. The cause of the ileus was detected in only 12.0% of the cattle examined. Ultrasonography is ideally suited for the visualisation of typical signs characteristic of ileus (intestinal dilatation and intestinal atony) but less so for determining the cause. The differences among the variables can be explained primarily by the type of ileus, the duration of illness, and the localisation of the intestinal ileus.

Animals doi: 10.3390/ani16182849

Authors: Jing Huang Teng Zhang Dongmei Xie Zhe Zheng Chuangye Yang Yongshan Liao Qingheng Wang Yuewen Deng

The offspring of domesticated Pinctada maxima exhibited various physiological and microbial adjustments to the complex and dynamic conditions of coastal environments. To support the restoration of P. maxima genetic resources and explore the molecular mechanisms underlying these phenotypic responses, we conducted a comparative analysis of the intestinal transcriptome and microbiota of wild parental and domesticated generations. Each sample generated an average of 43,600,525 clean reads, which were mapped to the P. maxima reference genome with mapping rates ranging from 63.91% to 79.48%, and a total of 3007 differentially expressed genes (DEGs) were subsequently identified. Gene Ontology analysis revealed that the DEGs were enriched in organic acid metabolism, and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that the DEGs were enriched in glycosphingolipid biosynthesis and xenobiotic metabolism via cytochrome P450. Microbiota profiling revealed significant compositional shifts at phylum and genus levels, with increased alpha diversity in F1; dominant phyla transitioned towards Spirochaetota and Bacteroidota, and functional predictions pointed to enhanced metabolic and immune capacities. Quantitative validated the up-regulation of immune genes (PmHR96h, PmIAP) and down-regulation of calcium-signaling genes (PmCaM, PmHSP90), consistent with RNA-seq data. Collectively, these coordinated transcriptomic and microbial alterations reflect a multifaceted host–microbiome adaptive response to nearshore conditions. Our findings provide valuable molecular markers and microbial indicators for selective breeding and health monitoring, offering a scientific basis for improving the resilience of P. maxima aquaculture under changing environmental conditions.

Animals doi: 10.3390/ani16182848

Authors: Leo J. Fleishman

Visual ecology explores how animal visual systems are related to light environments and important visual tasks. Anolis is a species-rich genus of small lizards that rely extensively on vision. Different species occupy habitats with different vegetation structure. The spectral properties of the habitats are similar, but total light intensities vary widely. They possess laterally positioned eyes with broad monocular visual fields, and a small region of binocular overlap toward the front. They possess a high-resolution central fovea associated with analysis of important images. There is a second, smaller, temporal fovea located where the visual fields of the two eyes overlap, which is associated with distance perception during prey capture. Color and brightness vision depends on four classes of single cones with different spectral absorbance curves, including one sensitive to the ultraviolet and a set of double cones. Most species studied possess similar sets of cones. Cone responses are modified by oil droplet filters, which are more variable among species and may play some role in adaptations to habitat light. Anoles communicate with a colorful expandable throat fan called the dewlap. Dewlap visibility depends on contrast with the natural background. Red dewlaps are most visible in bright, unshaded habitats. Yellow dewlaps are most visible in darker, shaded habitats. Visual displays include motion patterns of the head, body, and dewlap. The most highly visible movements are rapid up-and-down, start-and-stop patterns. These often occur at the beginning of visual displays and draw the attention of conspecifics to the displaying animal. Most Anolis species share similar visual-system properties. Differences in light environments interact with visual-system responses to influence the physical properties of communication displays and other visually-based behaviors.

Animals doi: 10.3390/ani16182847

Authors: Yunqing Guo Rongrong Zhang Tengfei Zhang Wenting Zhang Qin Lu Qiao Hu Qingping Luo

The increasing detection of Riemerella anatipestifer in chickens poses severe challenges to the poultry industry. However, the genomic differences among R. anatipestifer isolates from different poultry hosts remain poorly understood. Here, we investigated host-associated genomic differences among 158 R. anatipestifer strains isolated from different hosts in China, using pan-genome, phylogenetic, virulence factor, and antimicrobial resistance analyses. The core-genome phylogeny showed no apparent separation according to host, sampling period, or geographic origin. Conserved genes including groEL, tufA, and katB were detected in all strains, whereas tviB and ureG displayed host-associated distribution patterns. More than 40 putative antimicrobial resistance (AMR) genes were detected across 158 R. anatipestifer strains, indicating the presence of multiple putative AMR determinants. Among these, blaRAD-1, blaRASA-1, and dfrA36 exhibited host-associated distribution patterns. Pan-genome analysis further identified host-associated genomic differences in R. anatipestifer. These included genes contributing to cell-surface modification pathways mediated by pglH, and genes involved in toxin–antitoxin system-dependent stress regulation via parD1/parE1 and yefM-yoeB. Overall, our comparative genomic characterization of R. anatipestifer characterizes host-associated genomic differences and provides a basis for further investigation of their biological significance.

Animals doi: 10.3390/ani16182846

Authors: Mutsumi Furukawa Minami Yanagihara Miyuko Ohta Akihiro Kakuzen Nagomi Takeuchi Eiki Asai Ryota Hirakawa Jahidul Islam Tomonori Nochi

Type I (slow-twitch) muscle fibers differ in proportion among skeletal muscles, but their local spatial organization remains poorly characterized. This preliminary cross-sectional study investigated whether muscle-specific characteristics were reflected not only in the proportion of slow myosin-positive fibers but also in their clustering architecture. The longissimus, psoas major, semitendinosus, and triceps brachii muscles were collected from pigs at 4 weeks and 6 months of age. All six pigs were female, and live body-weight records were unavailable. Slow myosin-positive muscle fibers (slow fibers) were identified by immunohistochemistry, and adjacent positive fibers were defined as clusters. In pairwise tests at 6 months, the proportion of slow fibers was significantly higher in the psoas major than in the other three muscles, although the main effect of muscle across both ages was not significant (p = 0.0565). Slow fibers occurred as either isolated fibers or contiguous clusters in all muscles, but their clustering patterns differed markedly. Notably, the psoas major muscle had a significantly higher proportion of slow fibers in large clusters than the longissimus and semitendinosus muscles at 6 months of age. These findings suggest that muscle-specific characteristics are reflected not only in the proportion of slow fibers but also in their local spatial arrangement. Thus, clustering architecture may provide a complementary histological parameter for characterizing muscle-specific tissue organization in porcine skeletal muscle.

Animals doi: 10.3390/ani16182845

Authors: Amr Kchikich Jaber Maataoui Raoya Khachani Anass Ben Moula Nathalie Kirschvink Bouchra El Amiri Mehdi El Mokaddem Youssef Chebli Samira El Otmani Said Barrijal Mouad Chentouf

Growing concerns about antibiotics in semen extenders have driven interest in natural antimicrobial alternatives. The primary objective was to evaluate whether Thymus satureioides essential oil (EO) could replace antibiotics in a skim-milk extender while controlling bacterial growth and maintaining the post-thaw quality of Beni Arouss buck semen. Eighty ejaculates from eight bucks were pooled per collection and assigned to 12 treatments: EO at 0–0.05% (v/v), each with or without penicillin–streptomycin. After cryopreservation, extenders were assessed for oxidative balance, and semen was evaluated for sperm quality, lipid peroxidation, and aerobic bacterial load. Anti-oxidant activity increased and lipid peroxidation in the extender decreased as EO concentration rose whereas antibiotic-containing treatments showed higher lipid peroxidation. In semen, EO at 0.01–0.02% caused no significant changes in motility, viability, morphology, or membrane integrity compared to the control. Antibiotics better preserved sperm quality and reduced bacterial growth, but within 0–0.02% EO they were linked to higher lipid peroxidation. At ≥0.03%, EO showed significant toxicity, sharply reducing sperm quality while bacterial growth correlated negatively with motility, viability, and membrane integrity and positively with abnormalities. These findings suggest that oxidative stress may not have been the primary driver of sperm damage, as much of the measured MDA likely originated from extender lipids. In addition, significant EO × antibiotic interactions for total motility, abnormalities, and bacterial growth indicate that the response to EO depended partly on antibiotic supplementation. Overall, the tested EO concentrations did not reproduce the combined bacterial control and post-thaw sperm preservation observed with antibiotics and therefore did not support replacing antibiotics under these conditions.

Animals doi: 10.3390/ani16182843

Authors: David E. Samata-Flores Daniel P. Cáceres Apaza Mauricio Soto-Gamboa Juan C. Marin Contreras

The suri, or lesser rhea (Rhea pennata), is a flightless Andean bird facing multiple anthropogenic threats. Despite these pressures, its reproductive ecology remains poorly documented near its distributional limits. This study provides the first comprehensive evaluation of the breeding ecology, nesting distribution, and threats for wild lesser rhea populations at their northern geographic limit in Moquegua, Peru. Across four consecutive breeding seasons (2021–2024), we combined systematic surveys, kernel density analysis, camera trapping, and micro-behavioral nest monitoring. We identified 14 nesting sites comprising 193 nests: 18 old with eggshells remains, 110 old without eggshells, 53 nests under construction, and 12 actives. Kernel density analysis indicated a total nesting area of 195.8 km2, with a low density of 0.15 nests/km2. Nests were primarily established on moderately sloping hillsides utilizing high-Andean desert vegetation for camouflage. The reproductive period extended from mid-July to mid-December, displaying marked interannual variation in breeding phenology. Camera trap records and field encounters confirmed nest predation and interactions by a native predator guild composed of mammals (Lycalopex culpaeus, Puma concolor) and raptors, while free-ranging domestic dogs act as localized anthropogenic disturbance factors and linear infrastructure represents potential movement constraints. These findings establish an essential ecological baseline, highlighting that the long-term persistence of this marginal population depends on standardized monitoring to detect subtle demographic shifts and on strategic connectivity conservation across high-Andean landscapes.

Animals doi: 10.3390/ani16182844

Authors: Piotr Baranowski Wilhelm Grzesiak

Fluctuating asymmetry (FA) is commonly used as a phenotypic indicator related to developmental stability, reflecting variation in an organism’s ability to maintain bilateral correspondence during development. Although pelvic morphology has been extensively investigated in the domestic cat (Felis catus), little is known about fluctuating asymmetry of the pelvic girdle and its associations with biological variables such as age, sex, and body mass. This study quantitatively evaluated fluctuating asymmetry of the pelvic girdle in 45 domestic cats using 14 bilateral osteometric traits. Measurement repeatability was assessed using the technical error of measurement (TEM), and traits exhibiting significant directional asymmetry were excluded prior to FA analysis. An individual FA_index was subsequently calculated from the remaining traits and examined in relation to age, sex, and body mass. Six traits exhibited significant directional asymmetry and were therefore excluded from further analyses. Measurement error was low (%TEM: 0.34% to 2.20%) and consistently lower than the corresponding FA values, indicating that the observed asymmetry was greater than the estimated measurement error and was therefore unlikely to be explained primarily by measurement imprecision. The mean FA_index was 0.041 ± 0.032 and showed a positively skewed distribution. Adult cats displayed significantly higher FA_index values than subadults (p = 0.007, effect size: r = 0.40), whereas males also exhibited significantly higher FA_index values than females (p = 0.009, effect size: r = 0.39). No significant association was detected between FA_index and body mass (R2 = 0.014, p = 0.432). These findings demonstrate measurable fluctuating asymmetry of the pelvic girdle and indicate that its magnitude varies according to age group and sex within the studied population. The observed difference between subadult and adult cats is consistent with the use of FA as a phenotypic indicator related to developmental stability but does not demonstrate a decline in developmental stability with age. The observed asymmetry may also reflect cumulative functional and biomechanical influences acting on the pelvic skeleton throughout life.

Animals doi: 10.3390/ani16182842

Authors: Carla Zamora-Perarnau Mauro Malvè Patricia Sebastián-Marcos Rocío Fernández-Parra

Pulmonic stenosis (PS) is one of the most common congenital cardiac diseases in dogs. Although echocardiography and computed tomographic angiography (CTA) provide valuable diagnostic and anatomical information, the hemodynamic consequences of PS remain incompletely characterized. This study describes patient-specific pulmonary artery hemodynamics in dogs with and without PS using computational fluid dynamics (CFD). Contrast-enhanced CTA and echocardiographic data from five dogs with PS and five control dogs without PS were used to reconstruct three-dimensional pulmonary artery geometries and generate patient-specific CFD models. Pressure drop (Δp), blood flow velocity, mean wall shear stress (WSS), time-averaged WSS (TAWSS), oscillatory shear index (OSI), relative residence time (RRT), and pulmonary artery resistance were quantified and compared between groups. Dogs with PS exhibited markedly increased transvalvular pressure gradients, peak flow velocities, and pulmonary artery resistance, together with high-velocity jets and localized post-stenotic recirculation that were absent in control dogs. Mean WSS and TAWSS tended to be higher in the PS group, whereas OSI values were similar between groups. In contrast, RRT was significantly lower (p = 0.032) in dogs with PS. These findings demonstrate that patient-specific CFD enables detailed characterization of pulmonary artery hemodynamics in canine PS, providing information that complements conventional imaging. Although its clinical utility in veterinary medicine has yet to be fully established, CFD can provide valuable insight into flow behavior and disease pathophysiology; however, its ability to yield precise, clinically applicable quantitative values remains limited. This approach shows promise for future applications in the clinical evaluation of PS and other cardiovascular diseases, although further studies are warranted to determine its clinical relevance in veterinary medicine.

Animals doi: 10.3390/ani16182841

Authors: Seong Il Baek Yong-Woon Ryu Kang-Hee Im Kang-Woong Kim Jin Woo Park

To evaluate the overwintering performance of threeline grunt (Parapristipoma trilineatum), field experiments were conducted on 3000 subadults and 5000 juveniles reared in sea cages along the southern coast of Korea (SC group), and 600 subadults and 700 juveniles reared in inland tanks on Jeju Island (IT group), under thermally suitable overwintering conditions. Fish were sampled from January to April. The SC group maintained acceptable survival rates of 77.2% in subadults and 75.8% in juveniles. Body weight and standard length in the SC group did not significantly change during overwintering; condition factor and hepatosomatic index varied at specific sampling points. The SC group juveniles exhibited lower plasma glucose levels than the IT group ones. Overwintering in sea cages influenced body composition, characterized by reduced crude protein and increased crude lipid content. Plasma cortisol levels in the SC group juveniles increased significantly in January, whereas hepatic heat shock protein-70 levels showed no distinct changes. Principal component analysis revealed no clustering among groups. Physiological variations in the SC group reflected seasonal adaptation, evidenced by stable survival and no strong coordinated changes among physiological responses. These findings highlight the potential of threeline grunt to overwinter in sea cages along the southern coast of Korea.

Animals doi: 10.3390/ani16182840

Authors: Yongliang Zhang Keyuan Wang Yue Yang Nan Geng

Reliable dairy goat detection in barns is challenging because uneven illumination, railings, and animal overlap weaken RGB boundaries. We developed DRG-LiteStar-YOLO, a lightweight RGB-camera-compatible detector that integrates monocular pseudo-depth with RGB features without requiring a dedicated depth sensor. DA3-Small was used to generate relative pseudo-depth maps, which were fused with RGB features through reliability-guided fusion and boundary-enhanced multi-scale aggregation. The dataset comprised 1521 images of 134 lactating Saanen goats and 13,985 annotated instances. On the held-out test set, DRG-LiteStar-YOLO achieved a precision of 0.961, a recall of 0.941, an mAP@0.5 of 0.978, and an mAP@0.5:0.95 of 0.735. Compared with RGB-only LiteStar-YOLO, the proposed method improved mAP@0.5 and mAP@0.5:0.95 by 1.6 and 3.8 percentage points, respectively. The five-seed cumulative ablation further showed that the full configuration achieved 0.734±0.002 mAP@0.5:0.95 compared with 0.697±0.004 for RGB-only LiteStar-YOLO. The detector contains 4.20 M parameters and 12.60 GFLOPs and achieves 86.2 FPS on an RTX 4090 with precomputed pseudo-depth maps. These results demonstrate that reliability-guided pseudo-depth fusion improves goat localization under complex barn conditions while retaining a compact detector design. DRG-LiteStar-YOLO provides an effective perception framework for RGB-camera-based dairy goat monitoring and supports future counting, tracking, and behavior-analysis applications.

Animals doi: 10.3390/ani16182839

Authors: Tao Chen Meijuan Bai Lizhi Wang

γ-aminobutyric acid (GABA), an inhibitory neurotransmitter, plays an important role in regulating stress responses through modulation of the hypothalamic–pituitary–adrenal axis, neuroendocrine activity, antioxidant defense, and immune function. Therefore, this study evaluated the effects of dietary rumen-protected γ-aminobutyric acid (RP-GABA) supplementation on alleviating long-distance transport-induced stress in beef cattle. Forty-eight healthy Simmental bulls were randomly assigned to four treatment groups: CON, G-L, G-M, and G-H, receiving RP-GABA at 0, 300, 500, and 700 mg/kg of concentrate dry matter, respectively. The experiment lasted 53 days, including a 21-day pre-transport feeding period, a 2-day transportation period, and a 30-day post-transport feeding period. Samples were collected on experimental Days 21, 23, and 54. The results showed that long-distance transport impaired antioxidant status, immune function, and stress hormone secretion in beef cattle. The responses to different dietary RP-GABA supplementation levels varied among serum parameters. Long-distance transport reduced triiodothyronine and thyroxine (T4) concentrations, while the G-L group alleviated these reductions and maintained higher levels than the CON group after transport (Day 23). The G-M group enhanced glutathione peroxidase (GSH-Px) activity after transport (Day 23) (p < 0.001) and promoted the elevation of interleukin (IL)-4 levels both before and after long-distance transport (Days 21 and 23). The G-H group significantly decreased serum cortisol concentration after transport (Day 23), while reducing epinephrine levels and increasing glutathione peroxidase activity (all p < 0.001). Moreover, the G-H group further enhanced the increases in IL-4 and total antioxidant capacity before and after transport (Days 21 and 23), and increased IL-6 concentrations before and after transport (Days 21 and 54). Regarding dose-specific effects, the G-L group showed limited effects on stress-related hormones, immune function, and antioxidant capacity, whereas the G-H group enhanced some antioxidant and immune-related responses but had higher pre-transport stress hormone concentrations. In contrast, the G-M group exhibited a relatively balanced response across stress hormone levels, antioxidant status, and immune regulation, characterized by higher T4 (p = 0.003), GSH-Px (p = 0.001), and IL-4 (p = 0.006) concentrations and lower alkaline phosphatase (p = 0.005) and tumor necrosis factor-alpha concentrations. Therefore, under the conditions of this study, dietary supplementation with 500 mg/kg RP-GABA in concentrate may represent a potential level for maintaining balanced physiological responses in beef cattle.

Animals doi: 10.3390/ani16182837

Authors: Zhihui Huang Xin’an Wang Benhe Ma Ruiyu Deng Yunyi Gao Guohong Yu Chuanduan Ke Haihua Wang Aijun Ma

To distinguish the direct regulatory effect of nano-selenium on selenoprotein pathways from its indirect induction of glutathione metabolism, we measured the expression of eight genes representing these two core pathways (selenoo1, trnau1apl, selenop, and gpx1 for selenoprotein synthesis; mgst1, cha2, hagh, and glo1 for glutathione metabolism) in Paramisgurnus dabryanus at 20, 40, and 60 days of feeding, enabling the characterisation of short-, medium- and long-term transcriptional responses. Additive main effects and multiplicative interaction (AMMI) and genotype environment interaction (GGE) biplots were used to analyse the genetic basis of expression of the two types of selenium-related genes. The AMMI analysis of the expression of two types of selenium-related genes in P. dabryanus showed that, for selenoprotein-synthesis-related genes, the effect of selenium concentration decreased slightly and then gradually increased over time. For glutathione metabolism-related genes, the effect of selenium concentration decreased linearly over time. For both gene types, as time progressed, a clear antagonistic relationship was observed between the genotype effect and the genotype × selenium concentration interaction. GGE biplot analysis indicated that, overall, for selenoprotein-synthesis-related genes, the top two based on comprehensive evaluation of both stability and high expression were gpx1 and selenop. gpx1 may serve as a potential candidate for assessing nano-selenium bioavailability, selenium nutritional status, and antioxidant responses, warranting further validation. Overall, the top two glutathione metabolism-related genes in terms of comprehensive evaluation were glo1 and hagh. glo1 may be a potential indicator of nano-selenium-induced oxidative stress or glutathione metabolic load.

Animals doi: 10.3390/ani16182838

Authors: Bert Driessen Lore Pellens Béke Nivelle Kirsten Van Rossem Johan Buyse

Foie gras production is based on the force-feeding (gavage) of waterfowl to induce hepatic steatosis and subsequent liver enlargement. This practice has been widely debated due to concerns regarding animal welfare. This review provides a comprehensive overview of foie gras production, with a primary focus on the welfare implications of force-feeding in ducks, while contextualising historical and alternative practices involving geese. Evidence was evaluated across behavioural, physiological, pathological and health-related outcomes, together with procedural, housing and environmental factors. Endocrine findings are mixed, but repeated restraint and tube insertion can cause injuries to the upper digestive tract, while progressive feed administration produces marked liver enlargement, altered hepatic function and increased metabolic heat load. Reduced activity, locomotor impairment, lesions and elevated mortality have also been reported, although study designs and production methods and conditions vary. Alternatives under investigation include spontaneous fattening, microbiome-based interventions, cultivated-cell products and plant-based or processed analogues. Evidence concerning their welfare benefits, product quality, scalability and economic feasibility remains limited. Overall, the accumulated scientific evidence raises significant concerns regarding the welfare of force-fed birds and underscores the need for continued exploration of viable alternative production strategies.

Animals doi: 10.3390/ani16182836

Authors: Mengyu Wang Mingyang Zou Chenghao Zhou Wei Han

Southwestern local chicken breeds in China have long been recognized for their strong adaptability and disease resistance, shaped by unique selective pressures during domestication. In this study, we combined Fst and π ratio analyses to detect selection signatures from RAD-seq data of four southwestern local breeds, Red Junglefowl, and 17 other Chinese local breeds. We identified 460 candidate genes under strong selection potentially associated with domestication, affecting behaviors, physiology, reproduction, immunity, and adaptability. Importantly, 34 genes showed strong and unique selection signals when comparing southwestern breeds with other local breeds. Among these, CCR6, CD40LG, and CSF3R represent candidate genes that have been previously associated with immune processes, while MAPK10, FGFR4, and STAT3 represent candidate genes potentially related to disease resistance. Furthermore, HTR1B, HSPB9, and RYR2 represent candidate genes potentially associated with environmental adaptability. Notably, ALX1, the only gene detected across all comparison groups, may reflect morphological adaptation related to feeding efficiency. Our findings provide valuable insights into the genetic basis of unique adaptations in southwestern local chicken breeds and highlight potential targets for breeding and conservation.

Animals doi: 10.3390/ani16182835

Authors: Aipeng Mao Zexin Su Jieying Jing Junning Pu Xiaohong Guo Shufen Xue Jing Fu Jingyi Cai Gang Jia Gang Tian

Rapeseed meal (RSM) is a promising protein ingredient for animal feed; however, its utilization is limited by poor digestibility and the presence of anti-nutritional factors. In this study, we first investigated the effects of extrusion and fermentation on the structural characteristics and chemical composition of RSM. Subsequently, 18 growing pigs (6 pigs per treatment, 1 pig per replicate) were used. In period 1, pigs received diets with 20% RSM or fermented extruded rapeseed meal (FERSM) for nutrient digestibility determination. After a 7-day washout, the same pigs were re-assigned to 30% RSM or FERSM diets (6 pigs per treatment) for amino acid digestibility measurement. The results showed that extrusion combined with fermentation resulted in a more porous and loosened structure of RSM. Compared with RSM, FERSM exhibited a significant reduction in glucosinolate (GLs) content, whereas trichloroacetic acid-soluble protein (TCA-SP) content was significantly increased (padj < 0.001). Furthermore, pigs fed the FERSM diet showed significantly higher standardized ileal digestibility (SID) and standardized ileal digestible content of methionine (Met) than those fed the RSM diet (padj < 0.05). In conclusion, extrusion combined with fermentation, when wheat bran was included in the substrate, effectively modified the physicochemical properties and chemical composition of RSM and was associated with improved Met utilization in growing pigs.

Animals doi: 10.3390/ani16182834

Authors: Caijuan Tian Jixuan Zhu Xuanjian Chen Wanxin Ma Panpan Niu Peiyi Bian Huan Gao Yuquan Li

We previously found that period (per) may exert a positive regulatory effect on ovarian development under the modulation of circadian rhythms. This study further explores the function of the core clock gene timeless. Thus, we cloned the tim gene (Ec-tim) from the ridgetail white prawn, Exopalaemon carinicauda. The full-length Ec-tim cDNA sequence is 6102 bp in length and encodes a protein of 1543 amino acids with a conserved TIM domain. Quantitative real-time PCR (qPCR) analysis revealed that Ec-tim displays prominent eyestalk-specific expression in both adult and sexually mature individuals. Under different light–dark (L:D) cycles, Ec-tim exhibited regular oscillations under various L:D ratios within a 24 h period, and even under constant darkness (0 L:24 D) (p < 0.05), suggesting that tim is an endogenous clock gene. After RNAi-mediated silencing of Ec-tim, the transcript levels of key circadian clock genes, including per and cryptochrome 1 (cry1), were significantly downregulated relative to the control group (p < 0.01). In contrast, the mRNA level of the ecdysone receptor gene (EcR) was markedly upregulated (p < 0.01). These findings suggest that tim may regulate the expression of per and cry1 to form a trimeric complex, synergistically modulating circadian rhythms to further affect ovarian development in shrimp. Collectively, this study provides valuable references for further research on circadian regulation and reproductive development in crustaceans.

Animals doi: 10.3390/ani16182833

Authors: Daniele Zambelli Giulia Ballotta Diego Bucci Lucia Pagliuca Marco Cunto

Benign prostatic hyperplasia (BPH) affects more than 95% of intact male dogs, yet most remain asymptomatic until prostatic enlargement causes clinical signs. Standardised assessment of BPH severity is therefore needed to improve clinical decision-making and enable comparable research outcomes. This study aimed to investigate current diagnostic and therapeutic approaches to canine BPH and to develop and assess a web-based application implementing the Zambelli Symptom Index (ZSI), an established symptom-based scoring system for BPH severity. A voluntary questionnaire was completed by veterinary practitioners to assess current clinical practice, and agreement between the web-based and reference paper-based ZSI classifications was evaluated. The survey revealed considerable variability in the recognition of clinical signs, assessment of BPH severity, and selection of treatment strategies, despite broad agreement on the diagnostic value of ultrasonography and the need for further investigation of prostatic enlargement. The web-based ZSI showed very high agreement with the reference classification. These findings demonstrate that the ZSI can be implemented digitally without materially affecting severity classification. The web-based tool may improve the standardisation and reproducibility of BPH assessment in clinical practice and research. Further studies are warranted to evaluate interobserver variability, clinical validity, and responsiveness to treatment.

Animals doi: 10.3390/ani16182832

Authors: Insu Choi Beoul Kim You-Jeong Lee Hyo-Min Woo Jae-Woo Choi Man Hee Rhee Yong-Myung Kang Dongmi Kwak Min-Goo Seo

Rodent-associated Trypanosoma spp. are widely distributed in wild rodents, but their epidemiology in the Republic of Korea remains incompletely characterized. This study investigated the prevalence, spatiotemporal distribution, and genetic characteristics of Trypanosoma spp. in 1169 wild micromammals representing 12 taxa, including 1010 Apodemus agrarius, collected from 18 sites across the Republic of Korea between 2022 and 2025 using molecular methods. Trypanosoma spp. DNA was detected in 58 (5.7%) Apodemus agrarius; all other micromammal species examined were negative. Fifty-seven samples were identified as T. grosi and one sequence was designated as a T. lewisi-like genotype. No significant regional differences in prevalence were observed. Annual prevalence estimates were presented descriptively because the seasonal and geographic composition of sampling differed among study years. When the seasonal comparison was restricted to 2022, 2023, and 2025, prevalence was significantly higher in autumn than in spring. The single T. lewisi-like genotype showed 98.5–99.7% nucleotide identity with reference T. lewisi sequences and clustered within the T. lewisi clade. Because species assignment was based on a single genetic marker, the sequence was conservatively designated as a T. lewisi-like genotype. To the best of our knowledge, this study provides the first molecular evidence of a T. lewisi-like genotype in a wild micromammal in the Republic of Korea. These findings provide updated epidemiological information on rodent-associated Trypanosoma spp. in the Republic of Korea.

Animals doi: 10.3390/ani16182831

Authors: Jingmiao Yang An Li Zhenyi An Xiaowei Zhou Qiang Chen Lijuan Zhang Kai Feng Fen Zeng Xiaojuan Chen Bin Sun

Unlike mammalian erythrocytes, teleost erythrocytes are nucleated and transcriptionally active, yet their immune functions remain not fully understood. The leopard coral grouper (Plectropomus leopardus) is an economically important marine fish and is highly susceptible to Vibrio harveyi infection. In this study, we aim to investigate the immune role of erythrocytes during V. harveyi infection. Flow cytometry analysis showed that erythrocytes actively internalize V. harveyi in a dose-dependent manner. Transcriptome profiling of erythrocytes from control and V. harveyi-infected groups was performed to reveal the host immune responses after bacterial internalization. A total of 5995 differentially expressed genes (DEGs) were identified and significantly enriched in immune-related pathways, with the MAPK signaling pathway, apoptosis, and NF-κB signaling pathway as the three core pathways. Heatmap analysis further revealed transcriptional downregulation of MAPK kinases, apoptotic components, and cytokines (interleukins, chemokines, TNFs, and interferons). These findings provide transcriptional evidence for the immune involvement of teleost erythrocytes and offer insights into host–pathogen interactions during V. harveyi infection.

Animals doi: 10.3390/ani16182830

Authors: Cheryl Travers Robert Sparrow Angie Sassano Megan Frances Moss Chris Degeling

Robots are being developed to perform the day-to-day operational tasks of animal agriculture, affecting the way in which humans and farm animals interact. There is the prospect of automation increasing to the point where humans would hardly set foot on farms, which, aside from the consumption of animal-based products, could entirely disconnect humans and farmed animals from each other. Yet the literature on public attitudes towards the introduction of agricultural robots into the human–animal relationship is sparse. Here, we consider the public acceptability of introducing robots into the human–animal relationship in agriculture. Between 2023 and 2025, we conducted 12 dialogue groups and two community juries with members of the Australian public on the social and ethical impacts of automation in agriculture. We draw on insights from these empirical studies to highlight the underlying values that inform different attitudes towards robots taking on tasks previously performed by humans in animal farming. Across our empirical work, we consistently found three key factors influencing the willingness of members of the public to accept agricultural robots into the human–animal relationship: moral responsibility for farm animals; human intuition and empathy; and a natural life as a condition of animal welfare. Our data suggested that the removal of humans from all contact with farmed animals by adopting highly roboticized farming will likely be rejected by the Australian public as “a step too far”, largely due to its moral and social implications. We identify a set of conditions that are likely to be important to public acceptability of the use of robots in animal agriculture.

Animals doi: 10.3390/ani16182829

Authors: Yumeng Huang Yuting Guo Wei Feng Lijuan Gao Yiran Fu Jiale Bai Feng Liu

Microplastics (MPs) widely distribute in marine environments and generate severe hazards to marine biota, yet little is known regarding the toxic impacts of aged MPs on benthic invertebrates and their surrounding sediment microbiota. This work aimed to fill this research gap by exploring the comprehensive toxicity of aged polymethylmethacrylate microplastics (PMMA-MPs) toward Urechis unicinctus and its sediment habitat. A 21-day laboratory exposure trial was performed with two PMMA-MP treatments: environmentally realistic low concentration (10 μg/L) and extreme high concentration (1000 μg/L). Aged PMMA-MPs accumulated in the body wall and intestine of U. unicinctus, with markedly higher body wall MPs loads in the high-dose group (p < 0.05). High-concentration PMMA-MPs significantly raised intestinal total protein and triggered intense oxidative stress (p < 0.01), and the Integrated Biomarker Response value rose dose-dependently. Sediment microbial β-diversity was obviously reshaped (p < 0.05), with enriched pathogens including Klebsiella and Enterobacteriaceae under high MP exposure, while the worm’s intestinal α-diversity and core microbiome remained stable, showing host adaptability. This study reveals multi-level toxic outcomes of aged PMMA-MPs on benthic worms and their sediment microhabitat, offering fundamental data to evaluate the ecological risks of weathered MPs in marine benthic systems.

Animals doi: 10.3390/ani16182828

Authors: Rochelle D. Moore Dorsa Amirkhany Kelsey Clark Behrad Noudoost

Chronic neurophysiology studies in Rhesus macaques (Macaca mulatta) require implantation of cranial hardware including headposts, recording chambers, and fixation screws. A common complication is persistent picking, scratching and grooming around implant margins, which can result in skin retraction, chronic inflammation, infection, repeated surgical repairs, and premature removal of the animal from research studies. Existing implant-care protocols rely heavily on topical therapies that may inadvertently increase grooming behavior. We developed a removable carbon fiber 3D-printed cranial cap that attaches directly to the headpost, covering the implant margins while allowing airflow and rapid removal during recording sessions. Three macaques received the protective 3D-printed cap. One animal tolerated the device for 836 days, during which implant manipulation ceased, surgical repair frequency decreased, and an additional 1056 h of electrophysiological data were collected. A second animal successfully used the 3D-printed cap during postoperative healing for 61 days following implant revision surgery. One animal did not tolerate the 3D-printed cap, and it was removed. This case study suggests that a simple, low-cost 3D-printed cap has the potential to improve wound healing, reduce implant manipulation, enhance animal welfare, and prolong the functional lifespan of chronic cranial implants in appropriately selected animals.

Animals doi: 10.3390/ani16182827

Authors: Stefano Spada Gaia Pagani Johannes Herbel Bianca Lourdes Frehner Joana Rodrigues Carvalho Hélène Jainek Martina Gavezzoli Andrea Major Sebastian Patrick Arlt

Prostatitis is one of the most common prostatic disorders in dogs, although its ultrasonographic features remain poorly characterized. The aim of this retrospective study was to describe the ultrasonographic findings of canine prostatitis and prostatic abscessation and to investigate their association with clinical and laboratory abnormalities. Medical records of 35 client-owned dogs with cytologically or histologically confirmed acute prostatitis were reviewed. Clinical, ultrasonographic, laboratory, bacteriological, treatment, and follow-up data were collected. The most common ultrasonographic findings were inhomogeneous parenchymal echotexture, intraparenchymal cystic lesions, capsular edema, periprostatic steatitis, and asymmetry of the prostatic lobes. Prostatic abscesses were detected in 25 dogs and showed marked variability in size and morphology. Abscess diameter was positively correlated with body weight (p = 0.001; rho = 0.6) and negatively correlated with the percentage of neutrophils with toxic granulation (p = 0.02; rho = −0.4). Most dogs recovered following medical or combined medical–surgical treatment. Follow-up examinations demonstrated resolution of the inflammatory changes surrounding the prostate, whereas parenchymal inhomogeneous echotexture frequently persisted. These findings provide a comprehensive ultrasonographic description of canine prostatitis and support the use of ultrasonography as a valuable tool for diagnosis and follow-up.

Animals doi: 10.3390/ani16182826

Authors: John Sebit Benansio Gift Simon Damaya Idika E. Okorie Saralees Nadarajah

Wetlands support fisheries and other livelihoods but can also bring people into close contact with crocodilians. Human–crocodile conflict has been documented across Africa, Asia, Australia and the Americas, yet indirect economic costs, including damage to fishing gear, remain less studied than attacks on people or livestock. We analysed questionnaire data from 379 respondents in purposively selected crocodile-exposed fishing communities in the southern Sudd Wetlands, South Sudan. The study quantified seasonal patterns of Nile crocodile (Crocodylus niloticus) damage to fishing gear, reporting pathways, conflict-resolution perceptions, responses to damaged gear and likely annual damaged-net categories. Reporting pathways were defined as the social and institutional channels through which respondents said gear-damage incidents were communicated; handling responses were defined as actions taken after gear was damaged. As expected from seasonal variation in floodplain use, the water level and fishing activity, reported damage was concentrated from July to December, with September the being most frequently selected main damage month (157 respondents; 41.4%). Quarterly ordinal responses differed among seasons (Friedman test: χ2=721.54, df = 3, p<0.001). Cumulative link models that included both quarters and the broader survey area showed the largest odds-ratio contrasts for July–September and October–December, whereas broader-area contrasts were smaller, and their confidence intervals overlapped one. Reporting was dominated by informal and community pathways: fishing-camp leadership, family members and other fishers each reached 100% agreement, whereas wildlife authorities received no affirmative reporting. Respondents also expressed little confidence in formal conflict resolution: 100% agreed that crocodile-related gear-damage conflicts were difficult to resolve, and 0% agreed that government would compensate for damaged gear. Damaged nets were mainly abandoned and replaced; repair was rare (1.6%). The most likely annual damaged-net category was 1–5 nets/year/participant (275 respondents; 72.6%). Because the survey was purposive, the results should be interpreted as patterns among sampled respondents and locations, not as prevalence estimates for all fishers in the Sudd Wetlands or South Sudan. The findings indicate that seasonal gear damage, weak formal reporting to wildlife institutions and limited institutional confidence should be central considerations in crocodile-conflict management.

Animals doi: 10.3390/ani16182825

Authors: Yongqing Li Shihao Liu Juan Geng Weiwei Wu

The Hetian Gray Donkeys (China_HTQD) and Xinjiang Donkeys (China_XJD) are representative local donkey breeds in Xinjiang, China, that show significant differences in production phenotypes, such as body size and coat color. However, their genetic background and the molecular mechanisms influenced by artificial selection have not been systematically elucidated. This study integrated whole-genome data from 60 newly sequenced China_HTQD and China_XJD individuals, along with 155 global reference datasets from the NCBI database, to construct a modern donkey whole-genome variation set covering domestic and overseas populations (including Africa, West and Central Asia, and Europe). The results of genetic diversity analysis indicated that China_XJD has slightly higher genetic diversity than China_HTQD, while the China reference population (China_Ref) and European population (Europe) have the lowest genetic diversity. China_HTQD and China_XJD have greater genetic variation. Both PCA and NJ phylogenetic analyses showed that China_HTQD and China_XJD are genetically very close, clustering within the Chinese donkey population background, and exhibit clear geographic genetic isolation from overseas reference populations. ADMIXTURE analysis (with K = 2 showing the lowest cross-validation error) revealed that the two breeds share a highly similar genetic structure, with only minor differences in the proportions of specific ancestral components, while both are distinctly different from the other four reference populations. A whole-genome differentiation scan (Fst) showed limited overall genetic differentiation between China_HTQD and China_XJD populations, but significant differentiation peaks were observed in some genomic regions. Based on the joint screening of windowed Fst (Fixation Index) and XP-CLR (Cross-Population Composite Likelihood Ratio), this study identified two candidate genomic regions containing the TBX3 and ZFAT genes. Among these, TBX3 emerged as a strong candidate with a robust selection signal and a well-established role in body size regulation. In contrast, ZFAT showed a weaker signal, and its potential involvement in coat color remains speculative and requires further validation. In summary, this study not only clarified the close genetic relationship and the level of interpopulation differentiation between China_HTQD and China_XJD at the whole-genome level but also identified candidate genes potentially associated with phenotypic divergence, providing gene-level confirmation of TBX3 and the novel identification of ZFAT as a candidate for coat-color variation, thereby providing a scientific foundation for elucidating the genetic mechanisms underlying phenotypic differences in Xinjiang’s local donkey genetic resources.

Animals doi: 10.3390/ani16182824

Authors: Andrea del Rincon Michael J. S. Duggan Milda Baltrimaite Aoife O’Kane John Mark O’Leary Pieter A. J. Brama Clodagh Kearney

This prospective, randomised, ex vivo study investigated the effects of manual double-spin centrifugation protocols on the cellular and growth factor composition of equine platelet-rich plasma (PRP). Blood from 20 healthy horses was evaluated in two phases. In Phase A (n = 10), nine first-spin protocols were compared, and two were selected based on platelet enrichment and leukocyte reduction. In Phase B (n = 10), these were combined with nine second-spin conditions, generating 18 PRP preparations. Platelet (PLT), leukocyte (WBC) and erythrocyte (RBC) concentrations and PDGF-BB and TGF-β1 concentrations were measured. No significant differences in cellular composition were identified among first-spin protocols (p > 0.05). From Phase B, all final preparations achieved platelet enrichment and leukocyte depletion, with negligible RBC contamination. Second-spin force significantly affected PDGF-BB and TGF-β1 concentrations (p < 0.001). PDGF-BB increased from 2629 pg/mL (95% CI: 2111–3274) at 200× g to 3467 pg/mL (95% CI: 2784–4317) at 1000× g, while TGF-β1 increased from 13,364 pg/mL (95% CI: 11,286–15,824) to 16,998 pg/mL (95% CI: 14,355–20,127). TGF-β1 also increased with second-spin duration (p = 0.01). Platelet concentration was positively correlated with PDGF-BB (ρ = 0.604) and TGF-β1 (ρ = 0.691; both p < 0.001), whereas WBC concentration showed no significant association with either growth factor. Thus, both preparation methodology and donor-related variability influence equine PRP composition, indicating that protocol standardisation alone is unlikely to ensure biologically consistent PRP products.

Animals doi: 10.3390/ani16182818

Authors: Jiajun Zhang Fangyuan Xiong Mengyao Li Shijie Wang Xiaoya Wang Li Wang Yuansheng Zhu Miao Li Zechen E Xuemei Li Wanjie Cai Hongfei Li

Hydropower regulation can alter riverine habitats and potentially influence fish diet, gut microbiota, and nutritional characteristics. In this study, upstream and downstream populations of Hemibagrus guttatus (H. guttatus) were investigated in a dam-regulated reach of the Hongshui River associated with the Datengxia Water Conservancy Project using 12S rRNA and COI metabarcoding, 16S rRNA gene sequencing, and muscle nutritional analyses. Fish from the two habitats exhibited distinct dietary compositions, accompanied by significant differences in gut microbial diversity, community structure, and predicted microbial functions. Upstream fish harbored higher microbial diversity and greater abundances of Firmicutes-associated taxa, including several putatively beneficial bacteria, whereas downstream fish showed enrichment of Proteobacteria-associated taxa and functions related to environmental adaptation. Muscle composition also differed between habitats, with upstream fish containing higher moisture and crude lipid contents and downstream fish exhibiting higher crude protein content. These habitat-associated differences suggest that natural dietary resources and intestinal microbiota are closely linked to host nutritional characteristics. These findings highlight the importance of considering upstream–downstream habitat origin when evaluating the trophic, microbial, and nutritional characteristics of H. guttatus and provide a biological reference for the management and conservation of fish populations in dam-regulated rivers.

Animals doi: 10.3390/ani16182823

Authors: Lei Xu Xingzhi Zhang Yafang Li Quehui Tang Jiajia Ning Lianggen Wang Shuangshuang Liu Delian Huang Dapeng Wang Pinyuan Wei Feiyan Du Jinxia Peng

The tri-spine horseshoe crab, Tachypleus tridentatus, is an evolutionarily ancient marine chelicerate of considerable ecological and conservation importance in the Indo-West Pacific. Although several mitochondrial genomes have been reported, comprehensive comparative mitogenomic analyses encompassing all extant limulid genera remain limited. In this study, we sequenced and characterized the complete mitochondrial genome of an F1 Tachypleus tridentatus individual derived from wild-caught broodstock from the Beibu Gulf, northern South China Sea, and subsequently maintained at an aquaculture facility, using high-throughput sequencing, and performed an integrated comparative mitogenomic analysis across the three extant genera of Limulidae. The circular mitochondrial genome was 15,006 bp in length and contained the typical set of 37 mitochondrial genes, including 13 protein-coding genes (PCGs), 22 transfer RNA genes, and two ribosomal RNA genes. The newly sequenced Tachypleus tridentatus mitogenome showed highly conserved genome organization and nucleotide composition relative to previously published conspecific mitogenomes, confirming established mitochondrial characteristics of the species. The principal contribution of this study is the addition of a complete Beibu Gulf mitogenome and its integration with 12 additional complete mitochondrial genomes representing all extant limulid species, together with four mitochondrial markers, K2P genetic-distance analyses, and independent BI and ML phylogenetic reconstruction. The concordance among these mitochondrial datasets provides an integrated assessment of genetic divergence and mitochondrial phylogenetic relationships within Limulidae and expands the geographic representation of complete mitochondrial genomic resources for Tachypleus tridentatus.

Animals doi: 10.3390/ani16182822

Authors: Yikai Lan Zengqiang Yin Lin Zhang Quan Yu Jiahang Wei Fan Du Lei Chen Jun Yang Tao Tian Hongmei Li

Sebastes schlegelii is a commercially significant species in the northern Yellow Sea. Recently, Sebastes schlegelii has shown a reduced stock density and a shift toward smaller body sizes in the northern Yellow Sea area off Liaoning Province. To comprehend the population dynamics and to formulate management tactics, we established a mathematical formula describing the length–weight relationship, as well as the corresponding growth equations for the body length and body weight. The biomass of Sebastes schlegelii in the northern Yellow Sea area off Liaoning Province was evaluated using biological survey data from 2019 to 2022. The findings reveal that the length–weight formula is expressed as W = 4.98 × 10−5L2.8878 (R2 = 0.9108), with L∞ = 407.6 mm, W∞ = 1717.04 g, K = 0.21 a−1, and t0 = −0.65 a. The critical age was 3.34 years, and the stock biomass was 13,141.35 tonnes. The current exploitation rate of this resource is below the maximum yield, indicating sustainable utilization. A fishing closure period from April to June is considered suitable. A minimum landing size of 240.48 mm is recommended for Sebastes schlegelii.

Animals doi: 10.3390/ani16182819

Authors: Mahmut Şamil Şamlı Kemal Kırıkçı

This study investigated the effects of short periods of incubation during egg storage (SPIDES) on egg quality, hatching performance, and chick quality in Alectoris chukar eggs subjected to extended storage durations. A total of 800 eggs collected from a 60-week-old chukar partridge breeder flock were allocated to eight experimental subgroups in a 4 × 2 factorial design, with storage period (35, 42, 49, and 56 days) and treatment (control or SPIDES) as the main factors (100 eggs/subgroup). All eggs were stored at 9–12 °C and 75% relative humidity in a pointed-end-up position with 24 rotations per day for their assigned storage duration (35, 42, 49, or 56 days). In the SPIDES groups, eggs were incubated for 2 h at 37.7 °C and 59% relative humidity in 7-day intervals, commencing on day 28 of storage. As storage duration increased, Haugh unit, albumen index, and yolk index decreased significantly (p < 0.001), while yolk ratio (p < 0.05) and yolk-to-albumen ratio (p < 0.01) increased. The overall hatchability decreased at 49 and 56 days of storage, while the hatchability of fertile eggs was lowest at 56 days (p < 0.05). Early embryonic mortality increased significantly at 56 days of storage (p < 0.05). SPIDES had a limited effect on most egg quality traits, including Haugh unit, albumen index, and egg component weights, but significantly improved the hatchability and hatchability rate of fertile eggs, reduced early embryonic mortality, and enhanced the overall chick quality score (p < 0.05). These findings indicate that chukar partridge eggs exhibit considerable resistance to long-term storage; however, SPIDES represents a beneficial management strategy for maintaining embryonic viability when storage duration exceeds practical limits.

Animals doi: 10.3390/ani16182821

Authors: Xingbo Liu Ziqi Zhao Jianmin Yuan Lei Yan Wei Nie

This study investigated the effects of dietary nano-selenium (nano-Se) levels and Se sources on growth performance, blood parameters, meat quality, and tissue Se deposition in broilers. In Experiment 1, 480 broilers were assigned to five treatments (0, 0.10, 0.20, 0.30, and 0.40 mg/kg nano-Se) to assess the dose-dependent effects of nano-Se, with 0.20–0.22 mg/kg identified as the optimal dose through quadratic regression. In Experiment 2, 480 broilers received either a control diet or diets supplemented with 0.20 mg/kg Se from nano-Se, sodium selenite (SS), or two organic Se sources (Se-enriched yeast 1 and 2, SY1 and SY2). Supplementation with 0.20–0.30 mg/kg nano-Se improved (p < 0.05) antioxidant status, carcass yield, muscle redness at 45 min post-slaughter, and Se concentration in serum. At 0.20 mg/kg, nano-Se, SY1, and SY2 showed greater improvements in carcass traits, and antioxidant status than SS (p < 0.05), with comparable efficacy between nano-Se and organic sources. Nano-Se, organic Se sources, and SS increased Se deposition in serum and breast muscle compared to the control (p < 0.05). In conclusion, 0.20 mg/kg nano-Se is recommended for broiler diets, demonstrating superior bioefficacy to inorganic Se and comparable effects to organic Se sources.

Animals doi: 10.3390/ani16182820

Authors: Anna Breithaupt Jacquelyn Jacobs

Nationally, there is a growing population of shelter dogs that are experiencing longer lengths of stay as euthanasia rates decline. Well-documented shelter-specific stressors can induce physiological and behavioral changes correlated with immediate and future welfare concerns. Dogs vary in their responses to the shelter environment based on genetics, past experiences, and context, leaving some individuals unable to adapt. Those that struggle may display maladaptive coping behaviors perceived as less desirable by adopters, perpetuating the stress response cycle. To address welfare concerns, behavioral interventions, including enrichment and training programs, may mitigate stress; however, there remains a population that could benefit from pharmaceutical interventions by supporting individuals at a physiological level. Shelter resources vary significantly, and differences in training, experience, and clinical staff can create challenges to accessing anxiolytic medications. Most shelter behavior assessments focus on temperament testing to predict future behavior at home or identify individuals at risk for aggression, but few exist that can help identify generalized stress related to their current environment. There remains a need for a validated behavior assessment that measures coping ability in a clinically meaningful way to facilitate partnerships between shelters and veterinarians, addressing immediate shelter dog welfare concerns.

Animals doi: 10.3390/ani16182817

Authors: Brandie Morgan-Jack Martin L. Katz Grace R. Kick Joan R. Coates

Canine degenerative myelopathy (DM) is a progressive, adult-onset neurodegenerative disease, with similarities to superoxide dismutase 1 (SOD1)-associated ALS. Clinical signs of DM begin with asymmetric pelvic limb general proprioceptive ataxia and spastic paresis that progresses to flaccid tetraplegia and respiratory failure. Although pelvic limb muscle atrophy and peripheral neuropathy have been reported in DM, intramuscular SOD1 accumulation and muscle spindle pathology remain incompletely characterized. This study evaluated pathological changes in the tibialis cranialis muscle of Pembroke Welsh Corgis with early- and late-stage DM and age-matched unaffected controls. Histologic stains and immunohistochemistry were used to evaluate myofiber morphology, fiber type distribution, muscle spindle integrity, and SOD1 immunoreactivity. Compared to unaffected dogs, DM-affected dogs exhibited increased SOD1 immunolabeling, greater variability in myofiber size, altered myofiber shape and packing, and fibrosis. A progressive reduction in the percentage of type I myofibers was observed without evidence of fiber type grouping. These changes were evident during early disease and became more pronounced with disease progression. Muscle spindles exhibited late-stage disruption of intrafusal myofiber organization and marked increase in SOD1 immunoreactivty. The observed alterations suggest that the muscle pathology was, at least in part, intrinsic to the muscle, rather than being secondary to neurodegeneration. Our findings demonstrate early and progressive pathological changes in the tibialis cranialis muscle, expanding our understanding of skeletal muscle involvement in DM.

Animals doi: 10.3390/ani16182815

Authors: Jingzhao Zhang Kaiyou Deng Xueyu Zhu Weijun Wang Yanwei Feng Xiaohui Xu Zan Li Cuiju Cui Jianmin Yang Guohua Sun

Apostichopus japonicus is a temperate marine echinoderm whose growth and reproduction are strongly influenced by temperature. We compared adult sea cucumbers exposed to stepwise warming (10–12, 12–14, and 14–16 °C across three successive 20-day stages) with animals maintained at 15 °C. Growth indices, gonadosomatic index (GSI), gonadal histology, and energy-budget components were evaluated in females and males. The 60 d experiment used three replicate tanks for each treatment-by-sex combination. Compared with the constant 15 °C treatment, the stepwise-warming regime showed a higher female GSI at stage III; representative gonadal sections documented stage-related structural changes in both treatments. No significant treatment effects were detected for final wet weight, specific growth rate, daily growth rate, or feed conversion ratio. Excretory energy decreased during stages II and III, while the proportion allocated to gonadal growth increased. Stage-specific within-sex comparisons showed higher gonadal growth energy under stepwise warming in both sexes during stage II, while the clearest GSI difference occurred in females during stage III; treatment × sex interactions were not significant. The clearest differences between the two temperature regimes were observed during stages II and III (12–16 °C). The findings provide an integrative physiological basis for temperature management during broodstock conditioning of A. japonicus.

Animals doi: 10.3390/ani16182816

Authors: Xunyu Guo Lijia Li Shuhang Yin Zhibin Ban Wei Nie

This study aimed to determine the apparent metabolizable energy (AME), nitrogen-corrected AME (AMEn), and net energy (NE) values of corn for Arbor Acres (AA) broilers aged 12–14 and 26–28 days and to develop prediction equations. Ten corn samples with varying chemical compositions were selected. Test diets were formulated using the substitution method, in which corn replaced 40% of the energy-supplying components in a basal diet. A total of 396 male AA broilers at 9 days of age and another 396 at 23 days of age were used across the two metabolism trials. AME, AMEn and NE were determined using the substitution method combined with respiratory calorimetry. Correlations between energy values and chemical components were analyzed. Multiple linear stepwise regression in SPSS was used to establish prediction equations for AME, AMEn, and NE. The results showed that for 12–14-day-old broilers, AME, AMEn, and NE ranged from 13.84–15.66 MJ/kg, 13.39–15.35 MJ/kg, and 8.61–12.56 MJ/kg, respectively. For 26–28-day-old broilers, the corresponding values ranged from 14.62–16.30 MJ/kg, 14.42–16.05 MJ/kg, and 10.42–13.06 MJ/kg. Notably, NE values and AME/GE (gross energy) ratios were significantly higher in 26–28-day-old broilers than in 12–14-day-old broilers (p < 0.05), which increased by 9.33% and 3.96%, respectively. Correlation analysis showed that phytic acid (PA) was significantly negatively correlated with AME, AMEn, NE, AME/GE, and AMEn/GE in the younger group, whereas ash content was significantly negatively correlated with AME and AMEn in the older group. The prediction equations exhibited good fit, with the coefficient of determination (R2) of the best equation reached 0.97. These findings support the precise application of corn in feed formulation and provide a scientific basis for its efficient utilization in poultry production.

Animals doi: 10.3390/ani16182814

Authors: Jane Yu Craig Ruaux

Proteomics, the study of the structure and function of proteins, has been used to investigate candidate biomarkers and study the pathophysiology of various diseases. Biomarkers may be useful in diagnosing, monitoring and assessing treatment responses. This review provides an overview of proteomic techniques, describes the workflow and pipeline for biomarker discovery, and evaluates the current literature on proteomic studies in veterinary gastroenterology in dogs and cats. Current studies in this field are advancing our understanding of disease pathophysiology in conditions such as chronic enteropathy (CE). For biomarker development, proteomic research in veterinary gastroenterology is still in the early discovery phase. Substantial work on verification and validation is required before proteomic biomarkers can be reliably translated into clinical practice in companion animals.

Animals doi: 10.3390/ani16182813

Authors: Burcu Bayramoğlu Yahya Tuncay Tuna

Morphometric information on ambling horses raised in Türkiye remains limited. This study aimed to characterize the morphometric traits of ambling horses according to breeding origin, age, and sex and to provide baseline phenotypic information on the studied population. A total of 102 naturally ambling horses were evaluated, including indigenous horses raised in Bursa (n = 28), indigenous horses raised in Nazilli, Aydın Province (n = 65), and Afghan-origin horses raised in Bursa (n = 9). Morphometric measurements of the head, body, and limbs were recorded, and the effects of breeding origin, age, and sex were analyzed using a general linear model. Significant differences among breeding origins were observed for several morphometric traits (p < 0.05). Afghan-origin horses generally exhibited higher values for several body measurements, particularly withers height, body length, chest depth, chest width, and heart girth, reflecting morphometric differences within the studied sample. Age- and sex-related differences were also observed for several morphometric traits. The strongest correlation was observed between CH and BH (r = 0.920, p < 0.001), followed by WH and BH (r = 0.873, p < 0.001) and WH and CH (r = 0.860, p < 0.001). ITH was also positively correlated with BH (r = 0.825), CH (r = 0.808), and WH (r = 0.744) (all p < 0.001). Overall, this study provides baseline morphometric information on ambling horses from different breeding origins raised in Türkiye and contributes to the phenotypic characterization of the studied population. However, because the Afghan-origin group included only nine horses and the distribution of animals among some groups was unbalanced, the observed differences should be interpreted with caution. Further studies involving larger and more balanced populations are needed to confirm these findings.

Animals doi: 10.3390/ani16182812

Authors: Eleonora Uber Giacomo Riggio Marta Becchetti Maria Luisa Marenzoni Laura Menchetti Daniel Lolem Mesfin Mekonnen Moliso James Ocaa Carlo Ruspantini Pier Giorgio Lappo Silvana Diverio

Pastoralist communities in Karamoja maintain close, multifaceted relationships with animals from early childhood, yet little is known about how children conceptualise animal emotions and cognitive abilities. This pilot study explored these conceptions among Kasimeri Primary School students in Karamoja, Uganda, within the AICS All-In-One Project (AID 012590/04/1). A cross-sectional survey was conducted using a structured questionnaire administered to 326 students (mean age 13 ± 2 years). Data were analysed using descriptive statistics and generalised linear models (GLMs). Most children attributed a wide range of emotions to animals, with anger (79.6%) and happiness (78.2%) being the most widely recognised and disgust the least attributed (63.2%). Emotion ascription differed significantly by emotion type (p < 0.001) and species (p = 0.004), with significant interactions (p = 0.022). Species differences emerged for several emotions (all p ≤ 0.006), with ruminants and chickens receiving higher attribution than dogs, reflecting different species’ roles in pastoral life. Students from pastoralist households showed lower emotion scores than those with non-farmer fathers (p = 0.025). Cognitive ability attribution also differed significantly across species (all p < 0.001). These findings challenge Western-pet-centred frameworks, highlighting the need for culturally grounded, One Health-oriented educational approaches integrating local animal knowledge with formal welfare and health curricula.

Animals doi: 10.3390/ani16172811

Authors: Samuel Joseph M. Castro Roderick T. Salvador Romeo S. Gundran Janice S. Garcia Supitchaya Siriyakhun Jakkawat Pongsumpan Kannika Na-Lampang

African swine fever (ASF) remains a major constraint on swine production in the Philippines, but interpretation of its national trajectory is complicated by changes in surveillance effort and regional heterogeneity. We analyzed domestic-pig surveillance records from August 2019 through December 2023. After exact-row deduplication and validity screening, 50,852 laboratory records were retained, including 8929 ASF-positive records and 324,741 tested samples. Monthly positive-record counts, annual and surveillance-stream-specific sample positivity, geographic and regional trajectories, year-standardized seasonality indices, Bayesian Information Criterion-selected joinpoint regression, and sliding-window Estimated Dissemination Ratios (EDRs) were evaluated. Positive laboratory records peaked at 733 in September 2020. Annual sample positivity reached 20.6% in 2020 and declined to 3.9% in 2023, despite tested-sample volume increasing from 41,035 to 102,333. Regional peaks were staggered from 2020 through 2023, demonstrating asynchronous detected activity. September and October had the highest mean seasonality indices, although their 95% confidence intervals included 1. The selected joinpoint model identified breakpoints in June, September, and December 2020 and August 2022. A first-order autoregressive correlation structure (AR(1)) sensitivity model supported the 2020 changes but not the August 2022 change. The primary 14-day EDR identified 25 sustained expansion and 28 sustained contraction episodes; 7-day estimates were more volatile, whereas 14- and 21-day estimates were more concordant. These findings indicate a transition from intense nationally detected activity in 2020 to lower sample positivity but persistent, recurrent, and regionally heterogeneous activity through 2023. Integrating record counts, testing denominators, regional patterns, joinpoint regression, and EDR provides a reproducible surveillance framework, but does not estimate population prevalence, causal policy effects, or future transmission.

Animals doi: 10.3390/ani16172810

Authors: María Ezquerra-Durán María Esther Durán-Flórez Luis Javier Ezquerra-Calvo Pablo Cardenal-Morales Massimo Santella

Primary tracheal neoplasia is uncommon in domestic animals and humans, and most reported tracheal tumours are malignant. To the authors’ knowledge, this case represents the third confirmed case of tracheal leiomyoma reported in dogs. In human medicine, tracheal leiomyomas have also only been reported sporadically and account for approximately 1% of tracheal tumours. A 12-year-old spayed female Pit Bull was presented as an emergency because of an acute worsening of recurrent respiratory signs that were refractory to medical treatment. On initial evaluation, reduced mobility of both arytenoid cartilages was observed, and left arytenoid lateralisation was therefore performed. However, four days after the procedure, the dog continued to experience severe episodes of respiratory distress requiring orotracheal intubation. Subsequent bronchoscopy revealed an intraluminal tracheal mass occupying approximately 90% of the airway lumen. Because of this obstruction, an emergency temporary tracheostomy was performed caudal to the mass. Once the dog had been clinically stabilised, computed tomography was performed for surgical planning. Surgical treatment consisted of segmental tracheal resection and end-to-end anastomosis, with removal of a segment corresponding to 24% of the total tracheal length. Histopathological examination confirmed the diagnosis of tracheal leiomyoma. Complete surgical excision was achieved, supporting a favourable prognosis.

Animals doi: 10.3390/ani16172809

Authors: Ruthaiporn Ratchamak Khomsan Buathalad Maharach Matra Eakapol Wangkahart Wuttigrai Boonkum Vibuntita Chankitisakul

Bull semen cryopreservation is essential for preserving valuable genetics and supporting cattle breeding programs; however, freeze–thaw stress compromises post-thaw sperm quality through oxidative damage and membrane injury. This study evaluated the effects of L-serine supplementation on post-thaw semen quality, susceptibility to induced lipid peroxidation, and antioxidant enzyme activity in bull semen. Semen collected from four Brahman bulls was cryopreserved in a Tris-based extender supplemented with 0, 2, 4, or 6 mM L-serine. Post-thaw total motility, progressive motility, viability, plasma membrane integrity, MDA concentration, and the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) were assessed. Supplementation with 2 and 4 mM L-serine resulted in higher total motility and progressive motility, and greater plasma membrane integrity than the control and 6 mM treatments (p < 0.05), while viability was higher with 4 mM than with the control and 6 mM treatments (p < 0.05). MDA concentration following induced lipid peroxidation was lower in all L-serine-supplemented treatments than in the control (p < 0.05). SOD activity was higher with 2 mM than with the control and 6 mM treatments (p < 0.05), whereas GPx activity was highest with 2 mM, followed by 4 and 6 mM, with the lowest activity in the control (p < 0.05). CAT activity was unaffected by treatment (p = 0.1192). Overall, supplementation with 2–4 mM L-serine improved selected post-thaw sperm quality parameters and was associated with reduced susceptibility to induced lipid peroxidation and changes in apparent extracellular antioxidant enzyme activities. These findings support the potential of L-serine as a functional additive to improve bull semen cryopreservation.

Animals doi: 10.3390/ani16172808

Authors: Patcharaporn Kaewmong Piyarat Khumraksa Tatsawan Suttiboon Nuttapong Limusunno Domechai Kaewnoi Sasibha Jantrakajorn Peerapon Sornying Narissara Keawchana Pornphutthachat Sota Pimwarang Sukkarun Apinya Arnuphapprasert Watcharapol Suyapoh

Polyparasitism is frequently documented in cetaceans; however, the pathological consequences of concurrent multispecies helminth infections and their associated tissue responses remain incompletely understood. This study characterized multispecies helminth infections in two live-stranded dolphins from the Andaman coast of Thailand, a spinner dolphin (Stenella longirostris) and a pantropical spotted dolphin (Stenella attenuata), using integrated postmortem, parasitological, molecular, histopathological, and immunohistochemical approaches. Both dolphins harbored helminths involving multiple organ systems. Pulmonary nematodes were identified as Halocercus delphini, while larval and intestinal cestodes were identified as Clistobothrium sp. and Tetrabothrius sp., respectively. The most prominent pathological alterations occurred in the respiratory tract and included chronic pyogranulomatous bronchopneumonia, airway epithelial injury, fibrosis, and pulmonary congestion associated with Halocercus infection. Additional parasite-associated lesions included chronic enteritis, cholangitis with biliary epithelial injury and ductular reaction, pancreatic inflammation, and localized inflammatory responses surrounding larval cestode cysts in the blubber and testis. Immunohistochemistry demonstrated extensive interleukin-4 (IL-4) immunoreactivity in affected pulmonary and intestinal tissues, whereas interleukin-10 (IL-10) was undetectable. Nitrotyrosine and heat shock protein 70 (HSP70) immunoreactivity were widely distributed across parasite-associated tissues, while caspase-3 immunoreactivity showed tissue-dependent patterns. Collectively, these findings demonstrate that multispecies helminth infections in stranded dolphins were associated with multisystem pathological alterations accompanied by inflammatory, oxidative/nitrosative stress, cellular stress, and apoptotic responses. These findings highlight the potential pathological significance of polyparasitism in stranded dolphins and support consideration of parasite burden and associated tissue responses during cetacean postmortem investigations.

Animals doi: 10.3390/ani16172807

Authors: Ah-Young Kim BokKyung Ku Kyunghyun Lee

Mass companion animal mortality scenes pose major veterinary forensic and animal welfare challenges, particularly when remains are commingled, variably decomposed, or skeletonized. In March 2023, the remains of 1256 dogs and cats were recovered from a private residence in Yangpyeong-gun, Gyeonggi-do, Republic of Korea, after prolonged deprivation of food and water. Because whole-carcass enumeration was not feasible across the entire site, a standardized skull-based minimum number of individuals (MNI) approach was used to derive the scene-wide count by combining direct counts of intact, non-duplicated carcasses with counts of isolated skulls from disarticulated remains. A representative subset of 22 specimens, consisting of 16 carcasses submitted by the investigating police and 6 isolated skulls collected during the scene investigation, underwent detailed forensic examination including postmortem radiography, full necropsy, and toxicological screening. Body condition scoring was possible in 5 of the 16 carcasses, and all five scored 1 to 3 on a 9-point scale, consistent with severe to extreme emaciation. Toxicological screening of all 16 carcasses was negative for all tested substance classes. Gross and Radiographical examination of the 6 isolated skulls identified external traumatic fractures and scavenging punctures in 3 specimens, breed-typical persistent open fontanels without evidence of trauma in 2, and an indeterminate suture-associated lesion in 1. This report proposes a practical and reproducible veterinary forensic workflow for large-scale companion animal mortality scenes in which extensive decomposition prevents reliable direct enumeration. The combined use of skull-based MNI estimation, structured necropsy, whole-body postmortem radiography, toxicological exclusion testing, and standardized skull morphology assessment supports evidentiary documentation and courtroom prosecution in mass animal cruelty investigations.

Animals doi: 10.3390/ani16172806

Authors: Catalin Micsa Maria Roxana Turcu Ioana-Raluca Prunean Alexandru Diaconescu

Background: Uterine torsion is rare in domestic cats and, when diagnosis is delayed, is associated with high maternal and fetal mortality. Conventional B-mode ultrasonography cannot reliably distinguish viable from devitalised uterine tissue, and Doppler ultrasonography, despite its capacity to assess uterine, ovarian, and fetal perfusion, has not been systematically applied to feline uterine torsion. Case Description: A 4-year-old queen in late gestation presented with acute abdominal pain, lethargy, abdominal distension, and tachypnoea. B-mode imaging showed an enlarged, fluid-distended uterine segment with thickened walls. Colour, spectral, and power Doppler revealed an absent to markedly reduced arterial flow distal to the torsion, elevated resistive and pulsatility indices in the uterine and ovarian arteries, and the loss of peripheral perfusion. The absence of fetal heartbeats and umbilical flow confirmed fetal nonviability. Laparotomy confirmed the torsion of a congested, devitalised uterine horn, with asymmetric fetal loading (two kittens in the torted horn, one fetal resorption in the contralateral horn) as the likely biomechanical cause. Ovariohysterectomy was performed without derotation. Histopathology confirmed haemorrhagic necrosis, and haematology showed severe regenerative anaemia and marked leucocytosis, which resolved progressively after surgery. Conclusions: Doppler ultrasonography supplied vascular and fetal viability information unavailable from B-mode imaging alone, supporting ovariohysterectomy over conservative detorsion. Based on the feline case reports identified during the preparation of this report, this appears to be among the first descriptions of a systematic, multi-vessel Doppler protocol combining colour, power, and spectral modalities with resistive and pulsatility index measurements in feline uterine torsion; as this observation derives from a single case, it should be regarded as preliminary.

Animals doi: 10.3390/ani16172805

Authors: Yanan Zhong Runbo Luo Kexin Li Na Li Chenjun Jiang Zixuan Li Jiayan Huang Kongwang He Libin Wen Qi Xiao Sizhu Suolang

Although mixed viral infections are increasingly recognized as contributors to bovine diarrhea syndrome, diagnosing such co-infections remains challenging, particularly in high-altitude regions where surveillance is limited. In July 2024, a calf presenting with severe diarrhea and respiratory distress was identified on a cattle farm in Linzhi, Xizang, China. Using unbiased whole-genome sequencing (WGS) of the fecal sample, we assembled near-complete genomes of three distinct RNA viruses: two bovine viral diarrhea virus type 1 (BVDV-1) strains (subtypes 1v and 1q, designated BVDV-1/XZ87 and XZ87), one bovine enterovirus (genotype EV-E, designated BEV/XZ87), and one caprine parainfluenza virus type 3 (CPIV3/XZ87). The CPIV3/XZ87 genome exhibited 99.9% nucleotide identity to the goat-derived GS2017-2 strain from Jiangsu, China, raising the possibility of viral spread through livestock trade. Quantitative real-time PCR (RT-qPCR) confirmed the presence of all three pathogens (Ct values: 24.78 for BEV, 25.98 for CPIV3, and 31.28 for BVDV). This study provides the genomic evidence of a triple co-infection involving BVDV-1, BEV, and CPIV3 in Xizang. It illustrates the potential of WGS for unbiased pathogen detection in complex clinical specimens. The near-complete genomes generated here fill critical gaps in the virological surveillance of this epidemiologically under-sampled high-altitude region.

Animals doi: 10.3390/ani16172804

Authors: Emma Quade Madison Serrate Vageeswari Nathapettai Thanikaivelan Fnu Joshua Lei Zhai Frank Logiudice Linda J. Walters

Feral and domestic felines are continuously exposed to meso- (polymer length: 5–25 mm) and microplastics (polymer length < 5 mm) from environmental or household contaminants. Ingestion and inhalation are the primary pathways for internalization of plastic, non-plastic anthropogenic (e.g., rayon, AP), and natural cellulose particles. However, contamination information is limited for companion animals. This study focused on reproductive tissue of felines by obtaining samples from spay (ovary + uterus) and neuter (testes) procedures. The tissue was chemically digested and then analyzed via microscopy and Fourier-Transform Infrared Spectroscopy (FTIR) to detect and identify contamination. Overall, 71% of 100 individuals contained particles (meso- and microplastics, non-plastic anthropogenic particles, cellulose), of which 97.3% were fibers. Foreign particles were present in the uterus and ovaries of 100% of domestic females, while only 36% of feral males had particles in their testes. Overall, 25% of the FTIR-confirmed particles were plastic, 54% were AP, and 21% were cellulose. The most common plastic found was polyethylene terephthalate (PET), a common polymer used in packaging food and beverages, including manufactured cat food and bottled water. This study provides a baseline for understanding plastic and AP concentrations in the reproductive tissues of both feral and domestic felines.

Animals doi: 10.3390/ani16172801

Authors: Yuqing Mu Xuezheng Shang Jie Yang Xuezhuang Wu

Cyclic heat stress compromises egg production and antioxidant defenses in laying hens. The present study employed a short-term, 14-day cyclic heat-stress model to evaluate whether dietary supplementation with a chlorogenic acid-rich green coffee bean extract could mitigate these effects. Before initiation of the heat-stress experiment, all hens were maintained under thermoneutral conditions and received the basal diet without the extract. Following 1 week of adaptation and 4 weeks of thermoneutral feeding, 240 approximately 50-week-old Jingfen No. 8 hens were allocated to a thermoneutral control, a heat-stress control, or heat-stressed groups receiving 100 or 200 mg/kg extract containing 70% chlorogenic acid, with four independent replicates of 15 hens each (60 hens per treatment). Heat-stressed hens were subjected to a 14-day cyclic heat-stress challenge at 32–35 °C for 9 h/day. Heat stress decreased average egg weight and laying rate, increased serum malondialdehyde, and reduced total antioxidant capacity (p < 0.05). Both supplemented groups improved average egg weight and laying rate, reduced malondialdehyde and total cholesterol, and increased total antioxidant capacity relative to heat-stressed controls (p < 0.05). Supplementation altered hepatic expression of genes associated with proliferation, apoptosis, and autophagy. Exploratory metabolomics detected changes in the 100 mg/kg group but not the 200 mg/kg group relative to heat-stressed controls. The extract partially mitigated production and antioxidant impairments during cyclic heat stress.

Animals doi: 10.3390/ani16172803

Authors: Alexandro Fritzen Maisa Damo Ana Lara Amaral da Veiga Jean Carlos Martinotto Piaceski Letícia Lescano Neves Renato Santos de Jesus Suelyn de Oliveira Marques Miklos Maximiliano Bajay Luiz Eduardo Lobo e Silva Roger Wagner Gilnei Bruno da Silva Daiane Manica Margarete Dulce Bagatini Aleksandro Schafer da Silva

High-starch diets can impair gastrointestinal homeostasis in growing cattle by promoting dysbiosis, inflammation, and oxidative stress. This study evaluated the effects of a multi-active phytogenic blend containing Macleaya cordata extract, essential oils, live yeast, silymarin, bentonite, and beta-glucans on performance, inflammatory and oxidative biomarkers, volatile fatty acids (VFA) production, and microbiota composition in calves fed a high-starch diet. Twenty-eight 5-month-old Holstein males were assigned to a control diet (control group) or the same diet with addition of 15 g/animal/day of the additive (Phytogenic group) for 70 days. Calves fed with additives had greater weight gain and average daily gain (p = 0.05), whereas dry matter intake and feed efficiency were unchanged. The additive reduced total leukocytes and lymphocytes number, decreased of biomarker in serum as TNF-α, IL-1, C-reactive protein, ferritin, haptoglobin, creatine kinase, cholinesterase, TBARS, and reactive oxygen species. These animals of the Phytogenic group also have increased IL-10, IgA, IgG, total protein, and globulin concentrations compared to the control group. Ruminal and fecal VFA concentrations were higher in animals of Phytogenic group, including acetate, propionate, and butyrate. In addition, intake of additives increased ruminal microbial richness and altered beta diversity, indicating a marked shift in rumen microbial structure, while altering the relative abundance of specific taxa associated with high-starch feeding. These findings indicate that this phytogenic blend improves growth performance and exerts immunomodulatory, antioxidant, and microbiota-modulating effects, supporting its use as a nutritional strategy to mitigate the metabolic challenges associated with high-starch diets in growing cattle.

Animals doi: 10.3390/ani16172802

Authors: Chentong Wang Manyang Jin Yufan Li Yilong Lv Ju Chai Haitao Shi Xue Bai Yanling Huang Xi Wang

Mannan oligosaccharides (MOS) are functional prebiotics that selectively promote the proliferation of beneficial bacteria in the chicken intestinal tract. This study evaluated the effects of in ovo MOS injection on early growth, intestinal morphology, and cecal microbiota of broilers at 7, 14, and 21 days post-hatch (dph). In total, 700 Cobb 500 fertilized eggs received either no injection or 200 μL of PBS diluent, low (0.5 mg/egg), medium (5 mg/egg), or high (10 mg/egg) doses of MOS in the amnion. At hatch, approximately 280 male hatchlings from injection groups were distributed into 28 cages and raised until 21 dph. Our results indicated that MOS injection linearly increased feed intake and body weight during 0–7 dph and improved duodenal villus height and width, with no persistent improvements observed at older age. Moreover, microbiota shifts triggered by MOS were only observed at 7 dph. The MOS injection at 0.5 mg/egg reduced genus-level Sobs, and PCoA coupled with ADONIS analysis confirmed microbial separation across treatments (p = 0.017). MOS selectively enriched obligate anaerobic taxa, including Lachnospiraceae, Pseudoflavonifractor, and Collidextribactor (0.5 mg/egg), and Ruminococcus gauvreauii (5 mg/egg). Conversely, control-enriched genera (Enterococcus and Litchfieldia) were negatively correlated with body weight gain. In conclusion, in ovo administration of MOS at 0.5 mg/chick transiently improved the abundance of beneficial anaerobes, and injecting 10 mg MOS could improve villus morphology and growth performance in chicks in the first week, suggesting a potential role for MOS in shaping early microbial establishment in broilers.

Animals doi: 10.3390/ani16172800

Authors: Lu Jin Chunhua Zhang Shengli Li Chula Sa Min Nuo Ding Yang Wenting Li Le Fu Panliang Chen Yaxing Zhao Bo Wang Haizhou Sun

This study investigated the effects of graded dietary barley replacement for corn on growth performance, rumen fermentation, carcass traits, meat quality, fatty acid profiles, and the rumen microbiome in Arbas White Cashmere goats, and further applied untargeted muscle metabolomics to elucidate the metabolic mechanisms underlying meat quality alterations. Growth performance was monitored for all goats (n = 10 per group); rumen fermentation, carcass traits, serum biochemistry and meat quality were determined in six randomly selected goats per group (n = 6); and rumen microbiota profiling and muscle metabolomics were performed on three selected goats per group (n = 3). Partial barley substitution (33% and 67% of starch from barley) significantly improved final body weight and average daily gain (p < 0.05), whereas total replacement (100% barley starch) did not confer additional growth advantages. Moderate barley inclusion increased ruminal propionate concentration (p = 0.001) and enriched the fiber-degrading bacterium Prevotella, while total barley replacement markedly reduced rumen microbial diversity (Sobs, Chao1, ACE, and Shannon indices; p < 0.01) and depleted multiple fibrolytic and hydrogenotrophic genera, including Christensenellaceae_R-7_group, NK4A214_group, and Methanobrevibacter. The 33% barley group exhibited elevated serum total bile acids, FGF-19, and CYP27A1 levels (p < 0.05), suggestive of modulated bile acid metabolism, along with decreased glucose-6-phosphatase (p < 0.05), pointing to suppressed hepatic gluconeogenesis. Muscle metabolomics revealed that moderate barley substitution predominantly affected glycerophospholipid metabolism, with upregulation of PE(P-18:1/20:4) and PC(16:0/16:0), whereas total replacement induced extensive metabolic reprogramming characterized by downregulation of glycine and glutathione-related metabolites, indicative of compromised antioxidant defense. The 33% barley group also exhibited the highest muscular C18:3n-3 content (p < 0.01), while the 67% and total replacement groups showed increased C22:6n-3 deposition (p < 0.05). Correlation analysis confirmed that PE(P-18:1/20:4) and PC(16:0/16:0) were significantly positively correlated with ruminal propionate and ADG (r = 0.841–0.986; p < 0.05), and Prevotella abundance showed a strong negative trend with G-6-Pase (r = −0.771). Collectively, these findings demonstrate that replacing one-third of dietary corn starch with barley starch optimally balances productivity, rumen health, and meat quality through enrichment of Prevotella, enhanced propionate production, and modulation of bile acid metabolism, while excessive barley inclusion destabilizes the rumen ecosystem and triggers muscle oxidative stress, providing a theoretical basis for precision grain formulation in intensive cashmere goat production.

Animals doi: 10.3390/ani16172799

Authors: Sarah McCulloch Jan-Olaf Meynecke Jasper de Bie Madeleine J. Brasier

Humpback whales traditionally fast during migration and on breeding grounds, relying on reserved energy within blubber to sustain them. However, occurrences of feeding along migration routes and within breeding grounds have been documented for several populations, suggesting an intrinsic ability to adapt to changing conditions and prey variability. This systematic quantitative literature review yielded 77 publications, demonstrating that humpback whales feed outside their polar and sub-polar feeding grounds globally. We identified that the feeding documented within this review could generally be attributed to either opportunistic/supplementary feeding, or the identification of new or expansion of current feeding grounds. Key findings included substantial feeding behaviour in productive regions with high prey availability, including off eastern Australia, South Africa, and along the coasts of North and South America. Ontogenetic drivers may also encourage younger whales to forego migration to breeding grounds and remain along migratory routes during winter to exploit seasonally available prey. The definition of this feeding behaviour lacks specificity and may lead to underrepresentation in the literature, underscoring the need to investigate its prevalence and contribution to the behavioural budgets of these populations. Our study improves current understanding of the scope of feeding away from polar and sub-polar regions and encourages future work to further understand the drivers of the behaviour.

Animals doi: 10.3390/ani16172798

Authors: Tian Hua Simin Zhao Hao Bai Yulin Bi Wenming Zhao Guobin Chang

Cryopreservation plays a critical role in the long-term preservation of biological materials and has become an indispensable technology in biomedicine, agriculture, animal breeding, and biodiversity conservation. Despite substantial advances in cryobiology, the preservation of cells and tissues at ultra-low temperatures remains limited by cryoinjuries associated with ice crystal formation, osmotic stress, oxidative damage, and cellular dehydration. This review provides an overview of the historical development and fundamental principles of cryopreservation and critically summarizes the key factors affecting cryopreservation efficiency, including cryoprotective agents, antioxidants, osmotic balance, freezing and thawing protocols, and emerging ice-controlling materials. In addition, recent advances in the cryopreservation of diverse biological materials—such as sperm, oocytes, embryos, stem cells, and gonadal tissues from different animal species—are critically discussed, with particular attention to species-specific differences and the distinction between post-thaw survival and functional recovery. Finally, the current limitations of existing cryopreservation technologies and future research directions are highlighted to facilitate the development of safer and more effective preservation strategies for animal genetic resource conservation and biomedical applications.

Animals doi: 10.3390/ani16172797

Authors: Eun-Yeong Bok Chae-Rim Kwak Han Gyu Lee Duhyun Kim Eun-Do Lee Kwan-Woo Kim Ara Cho Younghun Jung Seog-Jin Kang Bock-Gie Jung

Coxiella burnetii is the causative agent of Q fever, a zoonotic disease of public health concern. Despite the recent expansion of the domestic goat industry in Korea, epidemiological data regarding C. burnetii exposure in Korean goat populations remain limited. This pilot study aimed to assess the serological distribution of C. burnetii exposure in the southwestern region of Jeonnam Province, the major goat-breeding region of Korea. Goat serum samples were originally collected for routine Foot-and-Mouth Disease monitoring in 2025. Using a two-stage random sampling approach, 92 samples were selected from 16 farms across five southwestern counties of Jeonnam Province. Specific antibodies against C. burnetii were detected using a commercial ELISA. The overall unadjusted apparent seropositivity was 44.6% (41/92). By county, seropositivity was 61.1% in both Haenam and Wando, 38.9% in Gangjin, 36.8% in Yeongam, and 26.3% in Jangheung. Furthermore, the overall herd-level apparent seropositivity was 68.8% (11/16). At the herd level within the surveyed farms, intra-herd seropositivity ranged from 0.0% to 100.0%, showing wide variation among the tested farms. The results indicate high serological exposure to C. burnetii within the surveyed southwestern counties. While constrained by a small sample size, these findings show that targeted longitudinal surveillance is required in this region. Expanding systematic monitoring efforts nationwide is recommended to establish effective veterinary and public health control measures.

Animals doi: 10.3390/ani16172796

Authors: Keqiang Li Teng Chen Dianzuo Yue Chuanlong Guo Sifeng Deng Xianglong Li

Early in ovo sex identification is important for improving hatchery resource utilization; however, sex-related optical differences are weak during early embryonic development and exhibit clear stage dependence. In this study, longitudinal visible–near-infrared transmission spectra were acquired from 1600 fertilized pink-shelled eggs of Bashang Long-tailed chickens across four independent incubation batches during incubation days 1–7. Before model development, 200 eggs were reserved as a fixed internal hold-out test set, while the remaining 1400 eggs were used for exploratory incubation-day and feature screening, five-fold cross-validation, and model comparison. Among all 21 dual-day combinations, D2 + D5 achieved the highest exploratory validation AUC of 0.951. The selected scheme required separate measurements on D2 and D5, with the final sex classification completed on D5. Further incorporation of the signed difference spectrum ΔS and relative change rate R increased the AUC to 0.965, demonstrating the incremental discriminative value of longitudinal developmental-change information. After the input scheme had been fixed, DSSF-Net achieved the numerically highest performance among the evaluated models under unified egg-level stratified five-fold cross-validation, with an accuracy of 0.915 ± 0.015, an AUC of 0.974 ± 0.012, and an F1-score of 0.915 ± 0.014. The final model achieved an accuracy of 0.915 and an empirical AUC of 0.966 (95% CI: 0.940–0.987) on the fixed internal hold-out test set. Complete four-round leave-one-batch-out validation yielded mean accuracy and AUC values of 0.846 ± 0.014 and 0.926 ± 0.012, respectively, indicating that the longitudinal spectral information from D2 + D5 maintained relatively stable discriminative performance across incubation batches within the investigated population. These results demonstrate the feasibility of longitudinal spectral modeling for early sex identification in fertilized pink-shelled eggs of Bashang Long-tailed chickens. Systematic offline validation was completed under laboratory conditions, including evaluation on the fixed internal hold-out test set and four-round leave-one-batch-out validation, supporting the effectiveness and batch-level stability of D2 + D5 longitudinal developmental spectra. Post hoc wavelength-level interpretation further identified 594–620 nm and 660–675 nm as two candidate spectral regions with relatively high predictive value. Collectively, these findings provide a reliable longitudinal spectral modeling framework for early non-destructive in ovo sex identification and establish an experimental and methodological foundation for wavelength selection, device development, and subsequent engineering validation of multispectral detection systems under hatchery conditions.

Animals doi: 10.3390/ani16172794

Authors: Sam Willems Amélie Canon Hanne Coppens Niels Demaître Nathalie Sleeckx Tomas Norton

Effective welfare monitoring in laying hens is increasingly challenged by growing flock sizes, declining farm numbers, and the practical limitations of assessor-based protocols under commercial conditions. Precision Livestock Farming (PLF) technologies offer opportunities to support large-scale welfare assessment, yet their application in laying hens remains predominantly limited to small-scale or prototype systems. This study addresses three complementary objectives aimed at informing future computer-vision-based PLF research and development in commercially housed laying hens. First, a systematic literature review was conducted to identify welfare-related categories monitored in laying hens between 2005 and 2025, the methods used to assess them, and the extent to which automated monitoring approaches have been applied. Second, outcomes of a TransRegional Expert Panel (TREP) within the OMELETTE project were synthesised to rank priority welfare challenges, evaluate the feasibility of different monitoring approaches, and compare expert perspectives with trends identified in the literature. Third, two pan-tilt-zoom (PTZ) camera setups implemented in semi-commercial aviary systems were described as proof-of-concept examples illustrating how multiple welfare challenges can be monitored using a single, multipurpose camera system. The literature review revealed a pronounced imbalance in monitoring frequency, with feather pecking dominating the literature while several other welfare challenges, including piling, disturbed sleep, and toe-pecking, remain comparatively underrepresented. TREP outcomes confirmed feather pecking as the highest-priority welfare challenge but also highlighted the importance of integrated monitoring approaches that combine time-intensive assessments, shorter checklists, and automated systems rather than relying on single indicators. Experts further considered the use of digital devices during routine barn inspections to be practically feasible. The PTZ proof-of-concept demonstrates how priority welfare challenges identified through both literature and expert input can be operationalised through automated, scheduled, and location-specific monitoring under commercial conditions. Together, these findings highlight the need for future PLF research to move beyond isolated measurements and small-scale trials towards integrated, cost-effective, and farm-specific welfare-monitoring systems that support adaptive, data-informed management strategies—for example, within a Plan–Do–Check–Act framework—and enable the development of digital standard operating procedures that generate actionable insights under real-world commercial constraints.

Animals doi: 10.3390/ani16172793

Authors: Zhannur Niyazbekova Cai-Yue Gao Nursultan Makhanbetuly Kuanysh Kassen Yuan Xu Bekzat Baimirzayev Huanhuan Zhang Zhadyra Muslimova Kanat Orynkhanov Yessengali Ussenbekov Fengting Bai Junyan Wang Hasan Baneh Yelaman Serikov Pilong Liu Yu Jiang

Understanding the genomic basis of environmental adaptation is essential for the conservation and genetic improvement of domestic camels. In this study, we investigated the population structure, genetic diversity, and genomic variation potentially associated with environmental adaptation of Kazakh dromedary and Bactrian camels using whole-genome sequencing. Whole-genome sequencing data were generated for Kazakh camels (15 dromedaries and 16 Bactrian camels) and integrated with 131 publicly available genomes representing camel populations from the Arabian Peninsula, Iran, Xinjiang, Inner Mongolia, and Mongolian wild camels. Population structure, genetic diversity, and genome-wide selection were evaluated using principal component analysis, ADMIXTURE, nucleotide diversity, linkage disequilibrium, runs of homozygosity, genomic inbreeding (FROH), and selection scans based on FST, θπ ratio, and XP-EHH. Population genomic analyses revealed clear differentiation between dromedary and Bactrian camels, whereas Kazakh camel populations exhibited higher nucleotide diversity (θπ = 1.307–1.551 × 10−3), and lower genomic inbreeding (median FROH: 0.037–0.056) than Arabian populations. Genome-wide selection analyses identified MC4R as the prominent candidate gene in Kazakh dromedaries and RYR1 as a prominent candidate gene in Kazakh Bactrian camels. Functional enrichment analyses highlighted pathways related to energy metabolism, thermogenesis, calcium signaling, skeletal muscle function, mitochondrial activity, and oxidative stress response. These findings provide new insights into genomic variation potentially associated with environmental adaptation in Kazakh camels and offer valuable genomic resources for future conservation, breeding, and evolutionary studies.

Animals doi: 10.3390/ani16172795

Authors: Daowen Qiu Yuwei Zhang Lirong Chang Yanchun Wang Zan Li Xiaokai Bao Xuebo Cui Cuiju Cui Guohua Sun Yanwei Feng Qiang Wang Xiaohui Xu Jianmin Yang Weijun Wang

Ammonia nitrogen represents a critical environmental stressor in aquaculture and exerts adverse impacts on the growth, reproduction and survival of shellfish. In the present study, diploid Pacific oysters (Magallana gigas) were exposed to ammonia nitrogen stress for 0, 6, and 48 h. We combined histopathological examination, antioxidant enzyme measurements, transcriptome profiling, and metabolome profiling to characterize time-dependent hepatopancreatic responses. Tissue injury became progressively more severe with prolonged exposure. Superoxide dismutase activity increased throughout the experiment, whereas catalase and glutathione peroxidase activities and malondialdehyde content showed an increase followed by a decrease, with no statistically significant differences in malondialdehyde content among the three time points. Transcriptome data revealed alterations in genes associated with ATP-binding cassette transporters, lysosomal function, endocytosis, and autophagy. Metabolic alterations were mainly associated with nucleotide, purine, pyrimidine, and sphingolipid metabolism. Cross-omics integration linked these molecular shifts to transmembrane transport, glutathione metabolism, sulfur-containing amino acid metabolism, and amino acid biosynthesis. The consistency between transcriptomic and metabolomic signals further highlights the central roles of transport processes, antioxidant defense, and metabolic adjustment under ammonia nitrogen exposure. These findings indicate that diploid Pacific oysters respond to ammonia nitrogen stress through coordinated regulation of oxidative stress responses, transmembrane transport, intracellular degradation and clearance, and metabolic reorganization. These findings provide insights into the adaptive mechanisms of bivalves exposed to ammonia nitrogen stress.