Search Results (696)

Mesenchymal stromal/stem cells (MSCs) are increasingly explored for immune-mediated diseases, yet standardized analytical readouts that capture coordinated immunomodulatory output across complementary secretory pathways remain limited. Here, we report the feasibility of an HPLC-based multi-analyte secretome characterization panel that quantifies two small-molecule outputs—adenosine and kynurenine—alongside two immunomodulatory proteins—interleukin-10 (IL-10) and transforming growth factor-beta (TGF-β)—in conditioned media from canine adipose-derived MSCs (cAD-MSCs). Canine immune-mediated hemolytic anemia (IMHA) was used as a disease context to motivate the selection of these analytes, given the pro-inflammatory cytokine environment characteristic of this condition. Three independent cAD-MSC lines were evaluated under baseline conditions and following cytokine stimulation with recombinant interferon-gamma (IFN-γ; 100 ng/mL) and tumor necrosis factor-alpha (TNF-α; 50 ng/mL), referred to herein as inflammatory priming or licensing. Conditioned media were collected at 72 h for metabolite analysis and 48 h for protein analysis, and quantified by HPLC using external calibration and peak integration. Across all three lines, licensing produced directionally consistent increases: mean adenosine increased 2.3-fold, mean kynurenine increased 3.1-fold, mean IL-10 increased 1.6-fold, and mean TGF-β increased 1.7-fold compared with unlicensed controls. Metabolite measurements for adenosine and kynurenine are reported with full chromatographic selectivity data; IL-10 and TGF-β measurements by reversed-phase HPLC with UV detection are presented as exploratory/semi-quantitative outputs and will require orthogonal confirmation (e.g., immunoassay) in future work. These findings are preliminary, derived from three independent donor lines with no comparator group, and are intended to support feasibility of the analytical framework rather than establish definitive performance specifications. Collectively, the data support the potential of a multi-analyte HPLC-based characterization panel to capture licensing-responsive secretory shifts across mechanistically complementary pathways, providing a foundation for expanded development and validation. Full article

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 (p ≤ 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. Full article

Neosporosis, caused by N. caninum, is an emerging protozoan disease responsible for significant economic losses in the global dairy and meat industries, primarily due to abortion in cattle. Dogs serve as both definitive and intermediate hosts and play a key role in the parasite transmission cycle. Currently, effective control strategies remain limited, partly due to insufficient information on infection status. In Vietnam, data on N. caninum infection are scarce and mainly limited to cattle and buffalo. In this study, an indirect enzyme-linked immunosorbent assay (iELISA) based on recombinant NcGRA4 protein was applied and evaluated for the detection of anti-N. caninum antibodies in dogs. A total of 142 shelter dogs from Hanoi, northern Vietnam, were tested to determine seroprevalence. The NcGRA4-based iELISA detected an overall seroprevalence of 28.87% (41/142), whereas the indirect fluorescent antibody test (iFAT) showed a lower prevalence of 14.08% (20/142), indicating substantial exposure to N. caninum among shelter dogs in this region. Using iFAT as the reference method, the NcGRA4-based iELISA demonstrated a sensitivity of 90.00%, a specificity of 81.15%, and an overall accuracy of 82.39%. These findings indicate that the NcGRA4-based iELISA is a suitable screening tool for seroepidemiological surveillance of N. caninum infections in dogs. Univariable and multivariable logistic regression analyses showed no significant associations between N. caninum seropositivity and the investigated variables, including age, sex, breed, and housing conditions. This study also provides the first serological evidence of canine exposure to N. caninum in Vietnam. Full article

This manuscript shows a clinical case of interest that was brought to the Teaching Veterinary Clinical Complex, College of Veterinary Science and Animal Husbandry, Jabalpur, Madhya Pradesh. Based on the clinical findings, infection with canine parvovirus type 2 (CPV-2) was a first-line diagnosis and later confirmed by a rapid antigen detection kit. Disease prognosis is influenced by viral virulence and host response, with early therapeutic intervention being critical for survival. The dog was given intensive supportive management including fluid therapy, antibiotics, antiemetics, antidiarrheals, antacids, and vitamin supplementation for six days. The dog became progressively better clinically, the gastrointestinal signs resolved completely, and the dog recovered fully. Full article

Background/Objectives: This study aimed to develop and evaluate deep learning models for the detection, localization, and classification of impacted maxillary canines, and to compare their performance with cone-beam computed tomography (CBCT) as the reference standard. Methods: This cross-sectional diagnostic accuracy study was conducted at King Abdulaziz University Dental Hospital to develop and validate artificial intelligence (AI) models for detecting and localizing maxillary canine impactions using panoramic and cone-beam computed tomography (CBCT) imaging data. A total of 641 panoramic ra and 158 CBCT scans were collected, of which 158 cases had matched panoramic–CBCT pairs for localization analysis. Images were annotated and validated by expert radiologists and orthodontists, with consensus review ensuring labeling reliability. Data augmentation expanded each panoramic and CBCT category to 500 samples for panoramic and 1000 samples for CBCT, resulting in 1935 panoramic and 5703 CBCT images after preprocessing and normalization. The datasets were divided into (training + validation) (80%) and testing (20%) subsets. MobileNetV2 architectures were used for classification, and whdiographsile, a ResNet-50–based Few-Shot Learning framework, enabled spatial localization of impacted canines. Models were trained using the Adam optimizer with a learning rate of 1 × 10⁻⁴ and evaluated using accuracy, precision, recall, F1 score, and area under the receiver operating characteristic curve (AUC). Cohen’s kappa and 95% confidence intervals were used to assess agreement between AI predictions and expert annotations. Results: Panoramic classification achieved 94% accuracy, demonstrating the highest performance in normal cases and reduced recall for bilateral impactions. The CBCT classifier achieved 98% accuracy across positional categories. Cross-modality prediction reached 93.5% accuracy, with strong agreement compared to CBCT (Cohen’s kappa = 0.91). Expert review confirmed reliable localization of impacted canines on both imaging modalities. Conclusions: Artificial intelligence applied to panoramic radiographs supports the detection, localization, and characterization of impacted maxillary canines with performance comparable to CBCT. This approach may enable lower-radiation decision support for clinical triage. Full article

The purpose of this study was to compare the retinal gene expression profiles in canines with Sudden Acquired Retinal Degeneration Syndrome (SARDS) and Cancer-Associated Retinopathy (CAR) and identify shared and distinct molecular pathways. Previously published SARDS and CAR canine retinal microarray data were used for the purposes of retinal transcriptomic pathway analysis, followed by KEGG and GO pathway enrichment analysis using DAVID and MetaCore tools. Gene expression patterns were analyzed to detect the most important signaling pathways. ProteinBERT deep-learning language model, and large language models (LLM-Grok 4, ChatGPT4o) were used for analytical prediction of possible drug targets. Both diseases showed significant upregulation in T-cell co-stimulation and complement activation pathways, including CD86, DLA-79, and C5AR1. Downregulated genes were enriched in pathways associated with visual perception and cardiomyocyte signaling. CAR exhibited upregulation of tumor-related chemokine signaling (e.g., CCR5, CXCR4), while SARDS showed pronounced enrichment in vascular inflammation pathways. Analysis of drug targets identified different classes of drugs, which could be potentially utilized for SARDS and CAR treatment. SARDS and CAR share immune-related molecular signatures but potentially differ in secondary mechanisms—vascular inflammation and endothelial activation in SARDS versus paraneoplastic mimicry in CAR. These data provide potential insight into the pathogenesis of SARDS as well as CAR, and identify potential diagnostic and therapeutic targets. Full article

Canine parvovirus (CPV) is a major pathogen causing severe gastroenteritis in dogs. Since its emergence, CPV has undergone continuous evolution, leading to the predominance of variants such as CPV-2a, CPV-2b, and CPV-2c. To characterize the genetic features and evolutionary trends of CPV-2 at a regional level, 775 fecal samples were collected from domestic and stray dogs with suspected CPV-2 infection in Shanghai between 2016 and 2025. The overall positivity rate was 23.2% (180/775); incidence was substantially higher in stray dogs (30.2%) than in domestic dogs (15.9%). Thirty-one CPV-2 strains were successfully isolated. Temporal analysis revealed a pronounced genotype shift: isolates from 2016 to 2020 were predominantly New CPV-2a, whereas CPV-2c became the dominant genotype from 2021 through 2025. Sequence analysis identified the polymorphism of VP2 gene and characteristic mutations F267Y, Y324I, N426E, Q370R and A440T in CPV-2c strains. A novel I447M mutation was detected in several isolates. Phylogenetic analysis showed that Shanghai isolates formed distinct clusters; CPV-2c strains were closely related to the Asian lineage. Structural modeling indicated that mutations at residues L87M, T101I, Y267F, A297S, G300A, Y305D, I324Y, Q370R, N426E, A440T, and I447M may alter the tertiary structure of the VP2 protein, potentially affecting antigenicity and receptor recognition. Collectively, these results demonstrate the complete genotype replacement of CPV-2 in Shanghai; CPV-2c is now predominant. Identification of the novel I447M mutation and structural analysis of key amino acid substitutions provide insight into CPV molecular evolution. These findings suggest that vaccines primarily based on older CPV-2 or CPV-2b genotypes offer suboptimal protection, highlighting the need for updated vaccine strategies targeting prevalent CPV-2c variants. Full article

Background/Objectives: Antimicrobial resistance (AMR) in companion animals is an emerging public health threat due to zoonotic potential and limited therapeutic options. Dogs with otitis externa may harbor multidrug-resistant (MDR) bacteria, including Escherichia coli (E. coli), Proteus mirabilis (P. mirabilis), and Enterobacter cloacae complex (E. cloacae complex), some producing extended-spectrum beta-lactamase (ESBL) or AmpC β-lactamases. This study aimed to assess the prevalence, AMR patterns, MDR occurrence, β-lactamase production, and co-infection profiles of these pathogens in canine otitis externa. Methods: Ear canal samples were collected from 592 dogs presenting clinical signs of otitis externa, with one sample per dog included in the analysis. Samples were collected from veterinary clinics in Timiș County, Romania, from 2022 to 2025. Samples were cultured on blood agar and MacConkey agar, followed by biochemical testing and MALDI-TOF mass spectrometry for bacterial identification. Antimicrobial susceptibility testing against 15 agents across six classes was performed using the VITEK^® 2 system. MDR and β-lactamase production (ESBL, AmpC) were determined according to CLSI 2018 veterinary guidelines. Co-isolation with bacterial and fungal species were recorded. Results: E. coli, P. mirabilis, and E. cloacae complex were isolated in 9.12%, 6.25%, and 1.2% of cases, respectively. E. coli exhibited the highest resistance to aminoglycosides (tobramycin 72.2%, gentamicin 61.1%) and full susceptibility to carbapenems. P. mirabilis showed the highest resistance to ampicillin (54%) and trimethoprim + sulfamethoxazole (46%), with complete susceptibility to carbapenems and fluoroquinolones. E. cloacae complex displayed universal resistance to cephalosporins but remained susceptible to non-cephalosporin β-lactams (piperacillin–tazobactam), carbapenems and aminoglycosides. MDR prevalence was 35.2% for E. coli, 18.9% for P. mirabilis, and 14.3% for the E. cloacae complex. ESBL production was detected in 13% of E. coli and 8.1% of P. mirabilis isolates, while all E. cloacae complex isolates were AmpC-positive. Co-isolations were common, primarily involving Staphylococcus pseudintermedius (S. pseudintermedius) and Malassezia pachydermatis (M. pachydermatis). Conclusions: MDR and β-lactamase-producing bacteria were identified in dogs with otitis externa, emphasizing the importance of routine antimicrobial susceptibility testing, targeted therapy based on local resistance profiles, and continuous AMR surveillance to prevent treatment failure and mitigate zoonotic risk. Full article

Canine babesiosis is a vector-borne disease of dogs with a worldwide distribution, presenting varying clinical signs depending on the host, parasite strain and climatic factors. Beyond companionship, dog meat serves as delicacy and is also used for zootherapy in some African communities. In this study, we collated and analyzed molecular biology-based diagnostic data on Babesia species of dogs in Africa in order to elucidate the epidemiological factors of the disease on the continent. Four Babesia species—B. rossi, B. vogeli, B. gibsoni and B. canis—were detected in Africa based on the results from 40 studies that involved the screening of 9435 dog blood samples from 83 study locations. Babesia rossi was the most commonly reported (aggregate detection rate = 7.7%) and was detected in samples from all the African regions except northern Africa. Babesia vogeli was the second most commonly reported (aggregate detection = 4.8%) and was detected in all of the African sub-regions. There were few reports of B. gibsoni (0.6%) in the southern and western African regions, and a single case of B. canis in an untraveled Nigerian dog. So far, there were no reports of Babesia coco, Babesia conradae or Babesia vulpes (Babesia annae, Babesia microti-like) in any of the African countries that have been confirmed by a molecular method. This study presents a synopsis of canine babesiosis in Africa, and provides an overview of common clinical signs, etiologies and risk factors that will serve as a quick guide to veterinarians to achieve timely tentative diagnosis. Full article

Cognitive dysfunction and neurodegenerative disorders represent an increasing clinical challenge in aging dogs and cats, while objective and minimally invasive biomarkers for early detection and disease monitoring remain limited. Tear film is a biologically active fluid reflecting both local and systemic processes and offers a practical, non-invasive source of potential biomarkers in geriatric veterinary patients. Proteomic analyses of canine and feline tear film have revealed a complex protein composition, including molecules involved in inflammation, oxidative stress, immune regulation, and cellular homeostasis—processes implicated in neurodegeneration. However, growing evidence from human and veterinary research emphasizes the importance of CNS-specific and mechanistically informative biomarkers, such as markers of axonal injury, synaptic degeneration, and glial activation, which may provide a more precise framework for interpreting peripheral proteomic alterations. This review summarizes current knowledge on tear film proteomics in dogs and cats and discusses its potential relevance to cognitive dysfunction and neurodegenerative processes. Particular attention is given to the integration of tear-derived proteins with validated blood and cerebrospinal fluid biomarkers, as well as to methodological challenges and future research priorities. With appropriate standardization and clinical validation, tear film proteomics may contribute to the development of novel diagnostic and monitoring strategies for neurodegenerative disorders in companion animals. Full article

Leishmania infantum is endemic in Portugal, where dogs are the main reservoir and human visceral leishmaniosis remains a public health concern. Increasing evidence indicates that cats are also susceptible to infection and may contribute to local transmission, although nationwide data remain limited. This study aimed to estimate the seroprevalence of Leishmania spp. infection in cats in Portugal and to identify potential risk factors. Serum samples collected for unrelated clinical purposes and submitted to a veterinary diagnostic laboratory between December 2024 and March 2025, representing all districts of mainland Portugal and the Azores, were analyzed. Anti-Leishmania spp. antibodies were detected using the direct agglutination test, and associations with explanatory variables were evaluated through multivariable logistic regression. The overall seroprevalence was 8.9% (96/1080; 95%CI 7.3–10.7). One cat from Terceira Island (Azores) tested seropositive (1/10). Region was the only independent predictor of seropositivity, with cats from the Algarve showing higher odds of infection than those from other regions (adjusted OR 1.97; 95%CI 1.24–3.13; p = 0.004). These findings demonstrate widespread feline exposure consistent with known canine and human hotspots, and detection in the Azores suggests possible local transmission, supporting the inclusion of cats in multi-host surveillance within a One Health framework. Full article

Advanced glycation end products (AGEs) such as Nε-carboxymethyllysine (CML) and Nε-carboxyethyllysine (CEL) are implicated in chronic disease processes in humans and may serve as biomarkers of dietary exposure and metabolic health. Urinary measurement of AGEs is of interest due to its non-invasive nature and relevance to biobanking and field-based sample collection; however, AGE stability in urine under common handling conditions has been poorly characterized. This study evaluates the short-term stability of CML and CEL in canine urine stored at room temperature (20 °C) for up to 168 h prior to −80 °C storage. Midstream free-catch urine samples from eight healthy dogs were aliquoted, stored at defined intervals, and analyzed in duplicate using liquid chromatography–mass spectrometry (LC-MS) with isotope-labeled standards. Results demonstrate minimal detectable changes in CML and CEL concentrations, as well as in the CML/CEL ratio, over the ambient storage period. Inter-replicate agreement is high, and regression and non-parametric analyses show no association between storage duration and analyte concentration. These findings indicate that urinary CML and CEL measurements may remain reliable despite delayed processing, supporting field-based sampling and retrospective analyses. Evaluation of additional AGE species and storage conditions will further inform best practices for sample handling in veterinary and comparative biomedical research. Full article

Canine distemper virus (CDV) affects both domestic and wild carnivores and is associated with a high mortality rate. The virus can cross species barriers, infecting a wide range of mammals, which raises concerns for both wildlife conservation and domestic animal health. During our study, we processed a total of n = 552 oral and rectal swab samples from n = 260 red foxes (Vulpes vulpes) and n = 16 golden jackals (Canis aureus). The samples were collected by the National Food Chain Safety Office (NÉBIH) as part of a Rabies monitoring programme from Hungary in 2024. We performed a Real-Time RT-PCR, followed by a CDV-specific amplicon-based sequencing method using Oxford Nanopore Technologies to obtain the complete genome. All golden jackal samples tested negative, while both oral and rectal samples of one red fox tested positive for viral RNA. From this positive sample, we were able to sequence a partial CDV genome. Based on phylogenetic analysis of the haemagglutinin gene, our CDV sequence was assigned to the Europe lineage, one of the endemic lineages in the continent, infecting both threatened and common animals. This finding highlights the ongoing presence of CDV in wildlife populations and illustrates the value of integrated monitoring systems. Full article

Background/Objectives: Following centuries of systematic eradication, grey wolf (Canis lupus) populations across Europe have experienced a significant recovery over recent decades, which leads to concerns regarding, among others, anthropogenic hybridization. In Greece, the genetic status of the wolf population is largely unknown to date. Here, we genetically monitor and test for wolf–dog hybridization events in a recently established wolf population in the Parnitha Protected Area, in close vicinity to the capital city of Greece. Methods: One hundred and twenty-four wolf scat samples were genotyped at 20 canine-specific autosomal microsatellite loci and compared to available reference tissue samples from wolves and free-ranging dogs. Results: A minimum of 31 unique wolf individuals were identified, structured into at least three packs. No wolf–dog hybrids were detected in the study area. To validate the accuracy of the microsatellite analysis, an ancestry informative 93-SNP panel was applied to non-invasive wolf DNA samples from the study area, confirming the absence of hybrids among them. However, a possible wolf–dog hybrid was detected among reference wolf samples collected in Northern Greece, where individuals with atypical morphological traits are observed. The estimated census population size was in accordance with concurrently obtained camera trapping data, while heterozygosity values were low. Conclusions: This research represents the first systematic effort in Greece to genetically monitor wolves recently established in a protected area. It highlights the need for targeted management strategies based on genetic data to ensure balanced long-term conservation of wolves in peri-urban areas. Full article

Human remains detection (HRD) canines are a class of odor detection canines trained with training aids representative of distinctive stages of putrefaction. This is necessary for canines to detect the whole spectrum of decomposition products, which is encountered in operational deployments. Understanding the definition of the cadaveric profile can help better train canine detection teams and assist with technological developments. This study aimed to (1) monitor chemical odor profiles utilizing two containment vessels, and (2) monitor two preservation methods on chemical odor profile changes as a function of freezing to thawing cycles. Instrumental analysis used solid phase microextraction- gas chromatography/mass spectrometry (SPME-GC/MS) for identification of volatile organic compounds (VOCs) from tissue/bone samples of pig carcasses at various decomposition stages. Samples were analyzed weekly for six (6) weeks to monitor chemical odor profiles as a function of time. Clear vials provided slightly better storage stability for the fresh stage compared to amber vials. However, amber vials were more suitable for the advanced decay and skeletal stages, helping to preserve the chemical odor profile. Regarding the preservation methods, a continuous preservation method portrayed better reproducibility of the original odor profile throughout the 6-week period, as depicted from higher Spearman correlation values. This study is the first to explore simulated training aids under freezing conditions, uncovering the dynamic and complex nature of odor over time. The results highlight that understanding these shifting odor profiles is essential for canine handlers aiming to optimize the realism and effectiveness of maintenance training. Full article