Animals最新文献

{"title":"CGS-BR: Construction and Benchmarking of a Respiratory Behavior Dataset for the Chinese Giant Salamander.","authors":"Dingwei Mao, Yan Zhou, Maochun Wang, Chenyang Shi, Yuanqiong Chen, Qinghua Luo","doi":"10.3390/ani16081272","DOIUrl":"10.3390/ani16081272","url":null,"abstract":"<p><p>The Chinese giant salamander (<i>Andrias davidianus</i>) is a nationally protected species in China, and its respiratory behavior serves as a key indicator of its physiological state, health status, and biological rhythm. However, research on intelligent monitoring of its respiratory behavior remains limited due to several challenges, including the species' nocturnal habits, resulting in low image contrast and poor quality in dark environments; extremely subtle breathing movements; and high-cost manual annotation, leading to a scarcity of high-quality annotated visual data. These factors severely constrain the application of deep learning techniques in this field. To support research on respiratory behavior monitoring in the Chinese giant salamander, this study constructs and releases the CGS-BR dataset, which is the first vision-based dataset dedicated specifically to respiratory behavior detection in this species. The dataset was collected under controlled simulated breeding conditions and consists of 1732 images extracted from 215 high-definition video clips. Following a standardized procedure, each complete respiratory cycle is manually annotated into four stages: head-up, diving, exhalation, and inhalation. To validate the effectiveness of this dataset, this study selects YOLOv8n as the baseline model, which balances detection accuracy, speed, and parameter count, enabling efficient giant salamander respiratory detection under limited resources. By comparing it with several representative models, we provide a reliable evaluation of the dataset's applicability. CGS-BR aims to provide fundamental data support for research on respiratory monitoring in the Chinese giant salamander, laying the foundation for subsequent applications in conservation management, captive breeding, health monitoring, and early disease warning.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"16 8","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13114148/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147809775","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Reducing Protein Content with and Without Yeast Probiotic Actisaf Sc 47 Supplementation in the Diet of Dairy Cow: Effects on Nitrogen Use, Digestibility, and Rumen Microbial Protein.","authors":"Nizar Salah, Brigitte Gestes, Pauline Ly, Axel Blancou, Kheira Hadjeba, Julie Schulthess, Julie Duclos, Eric Pinloche","doi":"10.3390/ani16081277","DOIUrl":"10.3390/ani16081277","url":null,"abstract":"<p><p>This study aimed to evaluate the effects of lowering crude protein (CP) levels in the diet of dairy cows alone or in combination with a yeast probiotic on nitrogen utilization, digestion, and rumen microbial protein synthesis. In total, six cows were included in a 3 × 3 Latin square design. Each study period consisted of 23 days of diet adaptation and 5 days for sampling and data collection. Cows were randomly assigned to one of the three diets according to protein level and supplementation: control diet with 16.5 CP%DM (CTR), a diet with 14.5 CP%DM without Actisaf Sc 47 supplementation (LCP), and a diet with 14.5 CP%DM with Actisaf Sc 47 supplementation at 5 g/cow/day (LCPActisaf). Reducing protein content from 16.5 to 14.5 CP%DM alone or in combination with Actisaf Sc 47 significantly reduced nitrogen intake. Regarding fecal nitrogen (FN), no difference was observed between the CTR and LCP groups. However, FN was significantly lower in the LCPActisaf group compared with that of the CTR group. There was a tendency for lower FN in the LCPActisaf group compared with the LCP group. Compared with CTR, urinary nitrogen (UN) was not impacted by LCP but tended to decrease with LCPActisaf (<i>p</i> = 0.1). Compared with CTR and LCP, supplementation with Actisaf Sc 47 increased nitrogen use efficiency (NUE) (<i>p</i> < 0.05). A tendency for higher NUE was observed in the LCP group compared with the CTR group (<i>p</i> = 0.07). Crude protein digestibility was similar between the CTR and LCP groups and tended to increase with Actisaf Sc 47 supplementation. Compared with CTR, neutral detergent fiber (NDF) digestibility was not impacted by LCP but increased by LCPActisaf (<i>p</i> = 0.05); LCPActisaf did not impact NDF digestibility compared with LCP. Organic matter (OM) digestibility was similar among treatments. Although there was no difference between the three groups on rumen microbial protein synthesis (RMP), supplementation with Actisaf Sc 47 increased RMP. Reducing protein content in combination with Actisaf Sc 47 appears to be a good strategy, enabling both the environmental footprint linked to nitrogen losses to be reduced, and fiber digestibility and rumen microbial protein synthesis to increase.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"16 8","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13113429/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147809735","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrating Environmental Drivers and Trophic Interactions to Predict Spatial Distribution of High-Risk Marine Organisms at Nuclear Power Plant Cooling Water Intake.","authors":"Yunlei Zhang, Xinyue Hu, Linquan Cao, Guize Liu, Changchun Song, Yuan Jin","doi":"10.3390/ani16081275","DOIUrl":"10.3390/ani16081275","url":null,"abstract":"<p><p>Marine organisms that episodically aggregate near coastal nuclear power plant water intakes pose a substantial risk to cooling water security. Predicting the spatial distribution of such high-risk species remains challenging because their occurrence is shaped not only by environmental conditions but also by complex trophic interactions. In this study, we model the habitat distribution of three high-risk nektonic species, Dotted gizzard shad (<i>Konosirus punctatus</i>), Japanese swimming crab (<i>Charybdis japonica</i>) and squid (<i>Loligo</i> sp.), in the cooling water intake area of a coastal nuclear power plant in eastern Liaodong Bay using generalized linear models (GLMs) and joint species distribution models (JSDMs). Based on summer surveys conducted in 2024-2025, we explicitly incorporated trophic linkages among target species, their prey, and predators within JSDMs. Model performance was evaluated using cross-validation based on AUC, RMSE, and coefficient of determination (R²). Our results indicate that water depth was the dominant environmental driver for all three species, while chlorophyll-<i>a</i> concentration and distance to the intake exerted species-specific effects. By incorporating interspecific trophic associations and environmental responses, JSDMs showed consistently improved predictive performance relative to GLMs, with approximately 1.5-fold higher R² values and 10-30% lower RMSE, while offering enhanced ecological interpretability. The models revealed strong positive associations between target species and both lower-trophic prey and higher-trophic predators, suggesting that top-down and bottom-up processes jointly regulate aggregation dynamics. This study demonstrates that integrating trophic interactions into species distribution modeling substantially improves predictions of high-risk marine species near coastal infrastructure and provides an ecological basis for proactive management of cooling water intake systems.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"16 8","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13113618/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147809741","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a Highly Sensitive ELISA for Detecting Antibodies Against a Novel Variant Avian Reovirus Based on Dual σC and σB Antigens.","authors":"Fuxi Zhao, Wanyi Zhou, Yilin Yuan, Qiuyun Peng, Weibin Wang, Weisheng Cao","doi":"10.3390/ani16081273","DOIUrl":"10.3390/ani16081273","url":null,"abstract":"<p><p>This study developed a dual-antigen enzyme-linked immunosorbent assay (ELISA) based on σB protein and genotype 5-specific σC protein of avian reovirus (ARV). First, σB and σC proteins were expressed and purified using recombinant technology. Through optimization of coating conditions, the optimal antigen combination was determined to be a mixture of the two proteins at a 1:3 molecular ratio (total concentration: 0.8 μg/mL). Key parameters of the indirect ELISA were optimized via checkerboard titration. Validation confirmed that the dual-antigen ELISA exhibited a sensitivity of 1:3200 against genotype 5 ARV-positive sera, with no cross-reactivity and a coefficient of variation of 2.9-8.6%, demonstrating excellent reproducibility. In application testing, the method specifically detected serum antibodies against genotype 5 ARV variant strains, achieving a 100% positive detection rate in experimental chickens within the first week post-challenge and effectively monitoring dynamic antibody changes in infected flocks. Furthermore, the detection rate for genotype 5-positive serum samples (100%) was significantly higher than that of a commercial kit (75%). This dual-antigen indirect ELISA overcomes the sensitivity limitations associated with conventional genotype 5 ARV detection methods and provides a reliable tool for epidemiological surveillance and infection monitoring.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"16 8","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13113393/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147809635","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Effect of Pellet Diameter on the Growth Performance, Nutrient Digestibility, and Intestinal Health of Piglets During the Creep Feeding Stage.","authors":"Lingao Kong, Fangxing Ou, Shuang Dong, Nan Zhang, Yongxi Ma","doi":"10.3390/ani16081260","DOIUrl":"10.3390/ani16081260","url":null,"abstract":"<p><p>This experiment evaluated the effects of pellet diameter on growth performance and intestinal health of piglets during the creep feeding stage. A total of 144 7-day-old suckling piglets (body weight of 2.2 ± 0.3 kg) were randomly assigned to four groups and fed the same formula as meal feed and pellets of 2 mm, 4 mm, and 8 mm in diameter, respectively. Each treatment consisted of six replicates of six piglets. The trial was divided into two phases by weaning time: 7-21 days (breast milk + creep feed) and 21-35 days (creep feed only). After the feeding trial, piglets from the meal feed group and the 8 mm pellet group were selected for slaughter and sampling. The results showed that before weaning, average daily feed intake (ADFI) increased significantly with increasing pellet diameter (<i>p</i> < 0.001). Post-weaning, piglets fed 8 mm pellets presented significantly higher final body weight (FBW) and average daily gain (ADG) than those in the meal group (<i>p</i> < 0.05). Apparent nutrient digestibility (ATTD) in pellet groups was significantly higher than that in the meal feed group and rose with increasing pellet diameter (<i>p</i> < 0.001). The organ indices of the stomach and large intestine in the 8 mm group of piglets were significantly lower than those of the meal group. The jejunal villus height (VH) in the 8 mm group showed a trend toward an increase (<i>p</i> = 0.066), and the ileal crypt depth (CD) was significantly lower (<i>p</i> = 0.004), with significantly higher digestive enzyme activities in the jejunum and ileum (<i>p</i> < 0.05). In the 8 mm group, the relative abundances of Bacteroidetes in the jejunum and Actinobacteriota in the cecum and colon increased, while those of Pseudomonadota decreased; jejunal microbial relative richness increased significantly, while the ileal microbial operational taxonomic unit (OTU) richness decreased obviously. In conclusion, pellets improved the growth performance of creep feeding piglets. Compared with meal, 8 mm pellets can significantly enhance intestinal health level and nutrient digestion and absorption capacity by optimizing intestinal morphology, boosting digestive enzyme activities, and improving flora structure.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"16 8","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13113964/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147809957","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transcriptional and Alternative Splicing Regulation of Autophagy and Vesicle Transport Pathways in Large Yellow Croaker Cells During Megalocytivirus Infection.","authors":"Zaiyu Zheng, Hongshu Chi, Xiaodong Liu, Xiuxia Chen, Ying Pan, Hui Gong","doi":"10.3390/ani16081259","DOIUrl":"10.3390/ani16081259","url":null,"abstract":"<p><p>Infection of the large yellow croaker (<i>Larimichthys crocea</i>) embryo cell line YCE1 with megalocytivirus strain FD201807 leads to accumulation of capsid-deficient viral intermediates within intracellular vesicles at 48 h post-infection (a phenotype associated with non-lytic egress), which coincides with the initial peak of viral genomic copies. To characterize the host molecular response during this critical stage, we performed time-course RNA sequencing at 24, 48, 96, and 144 hpi. Integrated analysis identified 6661 differentially expressed genes (DEGs) and 1138 differential alternative splicing (DAS) events affecting 892 genes, with DAS event abundance peaking at 48 h. DAS genes in autophagy and Golgi vesicle transport pathways, both integral to animal innate immunity, were significantly enriched exclusively at this timepoint, featuring novel mutually exclusive exon (MXE) isoforms in <i>gopc</i> (Golgi-associated PDZ and coiled-coil motif containing) and <i>rint1</i> (RAD50 interactor 1). Weighted gene co-expression network analysis (WGCNA) of DEGs identified <i>mapk9</i> (mitogen-activated protein kinase 9) and <i>map1lc3a</i> (microtubule-associated protein 1 light chain 3 alpha) as hub genes within modules enriched for autophagy-related functions. Separate co-expression analysis of DAS genes revealed <i>rnf5</i>, <i>rimoc1</i>, and <i>golga4</i> as hub genes, with <i>gopc</i> exhibiting only a single linkage to <i>rnf5</i>. These findings implied concurrent transcriptional and virus-induced host splicing regulation of vesicle-associated innate defense pathways and suggest that splicing-derived features may serve as potential candidates for diagnostics or prevention against megalocytivirus disease in <i>L. crocea</i>.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"16 8","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13113295/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147810098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Malic Acid Supplementation on Rumen Fermentation, Nutrient Digestibility, Performance and Carcass Traits in Lambs: A Meta-Analysis and Meta-Regression Considering Dietary Moderators.","authors":"Leonardo Tombesi da Rocha, Fernando Skonieski, Tiago Antonio Del Valle, Francine Basso Facco, Paola de Oliveira Selau, Kamily Pech Oliveira, Amanda de Vasconcelos Zucheto, Julio Viégas","doi":"10.3390/ani16081263","DOIUrl":"10.3390/ani16081263","url":null,"abstract":"<p><p>This meta-analysis and meta-regression evaluated how malic acid supplementation modulates rumen fermentation and its consequences for growth performance, nutrient digestibility, and carcass traits in lambs. Effect sizes (ES) were estimated using a random-effects model. Dietary composition was explored by meta-regression as a key source of heterogeneity, and subgroup analyses were used to compare free malic acid (FMA) and malate. Ruminal pH was not affected by malic acid supplementation. In contrast, total volatile fatty acid concentration increased with malic acid supplementation, particularly in studies using FMA. No effects were detected for propionate concentration, whereas acetate concentration increased (ES = 0.502; <i>p</i> = 0.036). A tendency toward a reduced ruminal acetate proportion was observed (ES = -0.683; <i>p</i> = 0.072). Malic acid supplementation tended (<i>p</i> = 0.057) to increase body weight gain (BWG; ES = 0.325) and final body weight (FBW; ES = 0.234). Malic acid supplementation did not affect carcass traits or overall nutrient digestibility. Meta-regression consistently identified fiber intake-related variables as major moderators of the effects of malic acid. Overall, the effects of malic acid supplementation on lamb performance appear to be primarily driven by its modulation of rumen fermentation and strongly conditioned by dietary context.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"16 8","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13113034/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147809801","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analysis of the Composition and Phylogenetic Relationships of the <i>Acanthosaura coronata</i> Complex Including Molecular Identification of Historical Specimens.","authors":"Natalia B Ananjeva, Maryia I Matsiushova, Anton O Svinin, Olga S Bezman-Moseyko, Luan Nguyen Thanh, Nikolai L Orlov","doi":"10.3390/ani16081261","DOIUrl":"10.3390/ani16081261","url":null,"abstract":"<p><p>The genus <i>Acanthosaura</i> is characterized by a high level of cryptic species diversity and is subdivided into several species complexes. The phylogenetic relationships within the <i>A. coronata</i> complex remain unresolved due to the presence of cryptic lineages and limited molecular data for several species. In this study, these relationships are clarified using a molecular genetic analysis that integrates newly collected field samples and historical museum specimens with previously uncertain identification. Three mitochondrial genes (<i>cyt b</i>, <i>COI</i>, and <i>ND2</i>) from samples, including fresh collections of <i>A. murphyi</i> from Phu Yen Province (Vietnam) and museum specimens from Vietnam and Myanmar, were analyzed. In addition, morphological characters of the examined specimens with diagnostic traits of known species were compared. Phylogenetic analyses confirmed the distinct species status of <i>A. murphyi</i> and enabled the taxonomic reassignment of previously undetermined museum specimens to this species. Specimens from Vietnam and Myanmar formed a single, well-supported clade, suggesting a broader distribution for <i>A. murphyi</i> than previously recognized. It is demonstrated for the first time that <i>A. murphyi</i> belongs to the <i>A. coronata</i> complex, together with <i>A. coronata</i> and <i>A. cuongi</i>, a result consistently supported by both genetic distances and phylogenetic tree topology.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"16 8","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13113640/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147809723","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of Supplementation with Dimethylglycine Sodium Salt on Immunity, Intestinal Tissue Morphology, and Antioxidant Function in IUGR Lambs.","authors":"Yuwei Wang, Mengfei Li, Lin Ma, Yurong Lin, Cheng Zhang, Zhiqiang Cheng, Yong Chen, Changjiang Zang","doi":"10.3390/ani16081258","DOIUrl":"10.3390/ani16081258","url":null,"abstract":"<p><p>This study investigates the regulatory effects of dietary supplementation with DMG-Na on growth performance, immunity, and intestinal development in IUGR lambs. A total of 45 lambs were used: thirty IUGR (3.10 ± 0.16 kg) lambs were randomly assigned to IUGR or IUGR + DMG-Na (0.1% in milk replacer from days 7-56) groups, with fifteen normal birth weight lambs as CON (4.32 ± 0.17 kg). At 56 days of age, eight lambs per group were slaughtered for sample collection. Compared to CON, IUGR lambs showed a significantly lower final body weight and average daily gain (ADG) (<i>p</i> < 0.01); IUGR also severely compromised intestinal structure, markedly decreasing villus height and villus height-to-crypt depth ratio across all small intestinal segments (<i>p</i> < 0.01); immune function was impaired, with highly significantly lower jejunal secretory IgA (sIgA) (<i>p</i> < 0.01); and antioxidant capacity was diminished, evidenced by reduced jejunal GSH, catalase (CAT) and glutathione reductase (GR) activities (<i>p</i> < 0.05) and increased jejunal MDA content (<i>p</i> < 0.01). Compared to IUGR, IUGR + DMG-Na group had highly significant increased final body weight and significant increased ADG (<i>p</i> < 0.01); it enhanced intestinal morphology, notably increasing villus height and villus height-to-crypt depth ratio in the duodenum and jejunum (<i>p</i> < 0.01); immune markers improved, with elevated jejunal sIgA (<i>p</i> < 0.05); and antioxidant status was restored, demonstrated by increased jejunal GSH and CAT activities (<i>p</i> < 0.05) and decreased jejunal MDA content (<i>p</i> < 0.01). In conclusion, DMG-Na effectively counteracted IUGR-induced deficits by promoting intestinal development, immunity, and antioxidant capacity, ultimately improving growth performance.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"16 8","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13113172/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147809050","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extracellular Vesicle-Associated miR-222-3p and miR-186-5p as Potential Hypoxic Markers in Canine Osteosarcoma: A Preliminary In Vitro Study.","authors":"Raffaella De Maria, Manuela Poncina, Sara Divari, Lorenza Parisi, Sonia Capellero, Luiza Cesar Conti, Eugenio Mazzone, Federica Fratini, Luca Aresu, Lorella Maniscalco","doi":"10.3390/ani16081265","DOIUrl":"10.3390/ani16081265","url":null,"abstract":"<p><p>The hypoxic microenvironment plays a critical role in the progression of canine osteosarcoma (OSA) by promoting different cellular responses, including the release of extracellular vesicles (EVs). Given the clinical aggressiveness of canine OSA, the aim of this study was to evaluate the miRNAome profile in EVs released in vitro by four canine OSA cell lines under hypoxic conditions. In particular, for this study we used two commercial canine osteosarcoma cell lines (D17 and D22) and two primary osteosarcoma cell lines obtained in our laboratory (Penny and Wall). D17, D22, Penny, and Wall cell lines were cultured under normoxic and hypoxic conditions (200 µM CoCl₂) for 24 h. EVs were isolated by size-exclusion chromatography and characterized by nanoparticle tracking analysis and Western blotting. miRNAs extracted from EVs were then sequenced and analyzed using bioinformatics approaches. The most representative miRNAs were identified and validated by qPCR using the miRCURY LNA miRNA PCR assay. miRNome profiling identified 233 miRNAs differentially expressed in EVs across all analyzed cell lines. Among these, 94 miRNAs were detected exclusively under hypoxic conditions. From this subset, 43 miRNAs were selected for further validation by qPCR. The qPCR results showed that miR-222-3p and miR-186-5p were significantly downregulated in the Wall cell line under hypoxia (<i>p</i> ≤ 0.05). TargetScan and pathway enrichment analyses demonstrated that miR-186-5p regulates target genes involved in different cellular processes. In human osteosarcoma, low serum levels of miR-222-3p are associated with poor prognosis, while miR-186-5p is recognized as a key hypoxia-responsive miRNA. Collectively, these results suggest the potential of EV-associated miRNAs as biomarkers in canine OSA and support their relevance in translational and comparative oncology.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"16 8","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13113573/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147809649","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}