Search Results (238)

Weaning represents a major developmental milestone for foals, especially when 4- to 7-month-old foals are abruptly separated from their dams. Studies have shown that the post-weaning period is associated with physiological and psychological stress, which may affect immune function. The present study aimed to describe and analyze the pattern of blood inflammatory biomarkers related to the innate immune system. A gradual foal–dam separation procedure was chosen, which included post-weaning social buffering by a familiar adult horse. Twelve Thoroughbred foals (six fillies, six colts) aged 4–6 months were enrolled and divided into a colt and a filly group, and after weaning, were kept as such. Blood samples were collected before and up to 7 days after weaning. Serum cortisol and cytokines (IL-2, IL-6, IL-10, TNF-α, IFN-γ) were quantified using equine-specific ELISA kits. Routine descriptive statistics were used to present the data, and inferences were drawn using a two-way repeated measures ANOVA. The level of significance was set at p < 0.05. Results showed significant differences in the concentrations of IL-10 (p < 0.001), IFN-γ (p < 0.01), and TNF-α (p < 0.01) on various days post-weaning. An apparent significant sex effect was not shown, but IL-10 peaked on day 3 in fillies. IFN-γ decreased significantly by day 5 in both sexes. Conversely, TNF-α levels increased significantly by day 5. No significant changes were observed for cortisol. Full article

African horse sickness (AHS) is a lethal vector-borne disease caused by African horse sickness virus (AHSV) and represents a major threat to equine health and the horse industry. In 2020, outbreaks of AHS caused by AHSV serotype 1 (AHSV-1) were reported in Thailand, increasing the risk of AHS introduction into China. Given the safety issues associated with currently available live attenuated AHS vaccines, the development of safer and more effective vaccination strategies is urgently needed. In this study, we constructed a recombinant fowlpox virus (rFPV) expressing the AHSV-1 VP2 protein as a candidate vaccine, designated rFPV-VP2. The recombinant virus was verified by PCR and Western blot analysis, which confirmed the successful expression of VP2. Preliminary immunization trials were conducted in both mice and horses, and immune responses were evaluated via an indirect enzyme-linked immunosorbent assay (iELISA) and immunofluorescence assay (IFA). The results revealed that VP2-specific antibodies were successfully induced in the serum of rFPV-VP2-immunized animals. Notably, serum from immunized horses showed specific reactivity with AHSV-1, confirming the induction of AHSV-1-specific immune responses. Therefore, these results demonstrate that rFPV-VP2 is a promising candidate vaccine for AHSV-1 and provide a scientific basis for the development of safer preventive strategies. Full article

Thyroid diseases in horses are rare and pose challenges for veterinarians due to the complexity of clinical and diagnostic approaches. This retrospective case series describes cases of equine thyroid disease in a referral hospital population. Files of horses examined for suspected thyroid disease at an equine university clinic between 2009 and 2024 were reviewed. Data from nine horses examined for suspected thyroid disease, including signalment, clinical presentation, laboratory results, imaging findings, fine needle aspirates, biopsies, diagnoses, and treatments, were summarised. Diagnoses included thyroid adenoma (n = 6), multiple thyroid cysts (possibly thyroid adenoma (n = 1)), non-thyroidal illness syndrome (NTIS, n = 1), and iodine deficiency goitre (n = 1). Diagnostic modalities varied, with, besides manual palpation performed in every patient, basal thyroid hormone measurement and ultrasonography being the most common. Low serum iodine concentrations were noted in four horses, yet iodine supplementation was inconsistently applied. Hemithyroidectomy was performed in four horses, enabling histopathological diagnosis (three out of four). Despite being rare, thyroid diseases in horses require consistent and thorough diagnostic approaches, including imaging, laboratory, and histopathological examinations. Improved screening tools and research could enhance the diagnostic accuracy and management of equine thyroid disorders. Full article

The aim of this study was to identify differentially expressed microRNAs (miRNAs) in serum and synovial fluid (SF) samples of control horses and those with osteoarthritis (OA) to identify potential candidates for biomarkers of disease. Total RNA was extracted from serum and SF samples of control (n = 4) and OA (n = 9) horses and sequenced. Differential expression analysis, pathway analysis and miRNA target prediction were performed. A group of six miRNAs (eca-miR-199a-3p, eca-miR-148a, eca-miR-99b, eca-miR-146a, eca-miR-423-5p and eca-miR-23b) was selected for validation in an independent cohort (serum, n = 46; SF, n = 88). The effect of clinical variables on miRNA expression was also assessed. Sequencing analyses found 43 and 23 differentially expressed miRNAs in serum and SF samples, respectively. Pathway analysis showed miRNAs were involved in inflammatory disease/response and associated with OA pathways. miRNA expression in serum was strongly associated with the horses’ workload, while age had a pronounced influence on miRNA expression in SF. Distinct patterns of miRNA differential expression were observed in serum and SF samples from horses with OA compared to controls. miR-199a-3p and miR-148a warrant further investigation as potential biomarkers of equine OA. Further characterization of these molecular changes could provide novel insights into the mechanisms of early OA. Full article

West Nile fever is a mosquito-borne zoonotic disease caused by West Nile virus (WNV), maintained in an enzootic cycle between avian hosts and Culex mosquitoes. While birds are the principal reservoirs, WNV also infects a wide range of mammals, including humans, horses, and wildlife species. In this study, we assessed WNV seroprevalence in wild ungulates, wild boars, golden jackals, and the invasive rodent nutria in Serbia. A total of 522 serum samples from wild animals were tested. Antibodies against WNV were detected across all tested species, with seroprevalence rates of 37% in wild boars, 11.9% in nutrias, 32.4% in golden jackals, 50.6% in red deer, and 9.1% in roe deer. Detection of antibodies in both adults and juveniles provides evidence of recent transmission during the study period. These findings confirm widespread circulation of WNV in Serbian wildlife and suggest that wild ungulates, carnivores, and invasive rodents may serve as useful sentinel species for monitoring WNV prevalence and geographic spread in natural ecosystems. Full article

Horses maintained under traditional management systems and dependent on natural forages are often exposed to seasonal and compositional variations that can affect metabolic homeostasis. This study examined associations between forage nutrient composition and metabolic–morphometric indicators in horses from four agroecologically distinct regions of northwestern Romania. Eighty-eight horses managed under semi-extensive rural conditions underwent clinical examination, body condition scoring (BCS), cresty neck scoring (CNS), and fasting blood sampling. Forage samples (n = 34) from daily rations were analyzed for fermentable carbohydrate content, while serum insulin, leptin, and adiponectin were quantified using validated equine-specific ELISA assays. Forage composition varied substantially among regions, influencing both endocrine and morphometric outcomes. Horses consuming carbohydrate-rich forages exhibited higher insulin (0.95–219 μIU/mL) and leptin concentrations (925–28,190 pg/mL), accompanied by elevated BCS and CNS scores, whereas adiponectin levels tended to decrease with increasing carbohydrate content. These findings demonstrate that naturally occurring variation in forage quality can significantly influence metabolic regulation in horses managed under low-input, traditional systems. Integrating forage nutrient evaluation with clinical and endocrine assessments provides a practical framework for identifying animals at risk of metabolic dysfunction and guiding nutritional strategies to mitigate the incidence of laminitis and related disorders. Full article

This study presents the first investigation of West Nile virus (WNV) seroprevalence among farmed horses in Kazakhstan. In 2024, a total of 368 serum samples were collected from horses across 106 settlements in 10 regions of the Republic of Kazakhstan. Using an enzyme-linked immunosorbent assay (ELISA), antibodies to WNV were detected in 32 horses (8.7%; 95% CI: 6.2–12.0%) from six regions. Among the seropositive animals, 26 (81.25%) were females and 6 (18.75%) were males, ranging in age from 1 to 19 years. No statistically significant association between sex and the presence of antibodies to WNV was found in any of the six regions. Significant differences between age groups were observed in Aktobe (χ² = 12.16; p = 0.002) and Turkestan (χ² = 4.20; p = 0.040). In the remaining regions (Almaty, Zhetisu, West Kazakhstan, and Atyrau), no significant age-related differences were recorded (p > 0.05). These findings confirm the circulation of WNV among horse populations in Kazakhstan and highlight the practical value and effectiveness of using horses as sentinel indicators for WNV surveillance. Full article

Mosquito-borne viruses like West Nile virus (WNV) and Usutu virus (USUV) present growing public health concerns, especially with climate change and expanding vector ranges. This study describes the development and validation of a duplex Real-Time RT-PCR assay targeting β-actin (ACTB) mRNA as an endogenous control and a conserved 92 bp region shared by WNV and USUV genomes. Degenerate primers for ACTB ensure RNA extraction quality and PCR performance while enabling simultaneous detection of both viruses. A total of 1002 mosquito pools collected in Piedmont, Italy, during the 2024 vector season under the National Surveillance Plan for Arboviruses (PNA), were tested. The assay showed 100% accuracy—ACTB mRNA was detected in all pools, and six pools tested positive for WNV or USUV (three each). Diagnostic specificity was confirmed on 40 horse and bovine serum samples. Sanger sequencing confirmed ACTB identity across multiple mosquito species. The assay also demonstrated reproducibility across different operators and thermocyclers. The limit of detection (LOD) evaluation showed that the assay is capable of detecting viral RNA at very low concentrations, confirming its high analytical sensitivity. The duplex RT-PCR here developed is a reliable, sensitive, and specific tool for arbovirus surveillance, combining pathogen detection with internal quality control of RNA extraction and amplification, thus improving early warning and rapid response to mosquito-borne disease threats. Full article

Disuse-induced bone loss during stall confinement and immobilization is a major concern in horses because it impairs recovery and increases susceptibility to further injury. Experimental models are needed to evaluate therapeutic options, but most available equine models rely on cast immobilization, which is technically demanding and may be associated with complications. This study aimed to assess a simpler and less restrictive model to induce a quantifiable decrease in bone density in horses. Six French Standardbred horses underwent eight weeks of stall confinement, with a wooden wedge fitted to one front foot to elevate the heels during the last four weeks. Bone density was assessed using computed tomography (CT) examinations of both forelimbs performed at the beginning (M0) and after the confinement period (M2). Serum markers of bone metabolism (CTX-I, osteocalcin, bone-specific alkaline phosphatase, and hydroxyproline) were analyzed monthly from baseline to 2 months post confinement. Statistical analysis used Wilcoxon signed-rank tests and mixed models as appropriate. Computed tomography revealed a significant decrease in bone density after confinement (p < 0.05), more pronounced distally in the wedge limb. CTX-I levels varied with physical activity. This model provides a practical and reproducible alternative to cast immobilization for inducing equine bone demineralization. Full article

Some correlations between serum Cu, Zn and Se and cytokines have been reported in humans. Especially, the Cu:Zn ratio corresponded with inflammation. To date, relationships between microelements and proinflammatory proteins are poorly understood in horses. The aim of the study was to evaluate whether Cu, Zn and Se may influence turnover of IL-6, IL-8 and tissue factor (TF) in breeding and working horses. Blood samples obtained from 66 horses were analysed. There were 37 pregnant broodmares of different breeds, 13 barren broodmares and 16 race Thoroughbred horses. Serum Cu, Zn and Se concentration was determined using the gas flame atomic absorption spectrometry (GFAAS) method. Plasma IL-6, IL-8 and TF concentration was determined by the ELISA method. A coefficient correlation was carried out to compare the values of microelements studied with IL-6, IL-8 and TF using Pearson’s test. The values of IL-6 correlated significantly positively with Se and Cu:Zn ratio, IL-8 correlated positively with Cu and Cu:Zn ratio and negatively with Zn, and TF correlated positively with Cu, Cu:Zn ratio and Se. The Cu:Zn ratio varies significantly between horses, with high values occurring in horses with high levels of proinflammatory proteins, which may indicate the presence of a subclinical inflammatory process. The high variability of TF in the studied groups gives hope for the use of its determination in laboratory diagnostics of horses. Full article

Mycoplasma hyorhinis is an important respiratory pathogen in swine, yet optimal culture conditions for high-yield propagation remain undefined. This study compared horse serum (HS) and chicken serum (CS) at graded concentrations (10%, 20%, 30%) for their ability to support in vitro growth of four clinical M. hyorhinis isolates (strains A, B, C, and D). Cultures were prepared in modified Friis medium, and growth performance was assessed by final titer (color changing unit, CCU/mL) and time-to-detection at 10² and 10⁴ CCU/mL. All media supported growth, but HS consistently outperformed CS in both yield and growth kinetics. The highest titers (10⁹ CCU/mL) and shortest detection times (3.6–6 days) were observed in 20% HS for most strains. Increasing HS concentration to 30% reduced yield for several strains, suggesting a concentration-dependent inhibitory effect. CS demonstrated limited but strain-dependent growth support, with comparable performance to HS for strain B at lower thresholds. These findings identify 20% HS as an optimal supplement for efficient M. hyorhinis cultivation, while highlighting the potential of CS as a cost-effective alternative under certain conditions, with implications for diagnostic reagent production and vaccine development. Full article

Background: Streptococcus equi subspecies equi (S. equi) is the cause of strangles, one of the most prevalent diseases of horses worldwide. The disease is characterised by fever and the formation of abscesses in the lymph nodes of the head and neck, which can restrict the airway. A multicomponent subunit vaccine, Strangvac, has been shown to effectively reduce clinical signs of strangles and to reduce its incidence. Objective: The aim of this study was to determine the immune response against the immunoglobulin-cleaving endopeptidase IdeE, a key protective component within the vaccine and the ability of antibodies to neutralize the proteolytic activity of IdeE. Methods: An in vitro assay was developed to measure the functional inhibition of recombinant IdeE by horse sera pre- and post-vaccination. The IdeE-neutralising titres were compared to the corresponding IdeE-specific antibody titres measured by iELISA (indirect Enzyme-Linked Immunosorbent Assay). Results: A significant IdeE-specific antibody response in blood serum collected from ponies was induced after Strangvac vaccinations. Concomitantly, significant increases in the neutralising activity of IdeE occurred, persisting for at least 12 months post-second vaccination. IdeE-neutralising activity was further increased significantly after a third vaccination, even when the third dose was administered 12 months after the second dose, demonstrating that immunological memory to the vaccine persisted for 12 months. There was a significant correlation between the IdeE-neutralising activity of blood sera and the level of IdeE-specific antibodies. Conclusions: These data provide insights into one potential mechanism by which this vaccine protects Equids against or during S. equi infection. Full article

Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne zoonotic pathogen of growing public health concern in southeastern Europe. This study provides the first serological evidence of CCHFV circulation in Croatia, based on testing 1473 serum samples from farm and companion animals, including sheep, horses, cattle, goats, dogs, and cats. A total of 109 samples (7.4%) tested positive for CCHFV antibodies using a commercially available enzyme-linked immunosorbent assay (ELISA) kit. The highest seroprevalence was recorded in sheep (28.3%), followed by horses (4.3%) and a single cat (0.5%), with no antibodies detected in cattle, goats, or dogs. Almost all seropositive animals originated from coastal and subcoastal Croatia, where Hyalomma ticks are present. Only two seropositive cases were detected in continental areas. Sheep samples from several farms in Zadar County showed intra-farm seropositivity rates of up to 85.7%, suggesting localised virus circulation likely influenced by vector distribution and farm-level practices. No viral ribonucleic acid (RNA) was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR), consistent with the transient nature of viremia in most animal hosts. These findings confirm the silent circulation of CCHFV in Croatia and reinforce the need for targeted, regionally adapted surveillance strategies that integrate multiple hosts and support early warning systems aligned with the One Health concept. Full article

This study investigated the presence of antibodies to human adenovirus type 36 (HAdV-D36) in horses with different metabolic statuses, including normal, overweight, and those diagnosed with equine metabolic syndrome (EMS). A total of 151 serum samples were tested, of which 47.6% were positive for anti-HAdV-D36 antibodies. Although the horses were confirmed to be susceptible to HAdV-D36 infection, there was no significant correlation between infection and blood glucose or cholesterol levels. However, the triglyceride levels showed significant differences—they were particularly elevated in the seropositive horses with EMS. These findings suggest that the virus may act by a different mechanism in horses than in other species, and highlight the need for further research to understand its role in horses. Full article

Equine herpesvirus 1 (EHV-1) and Equine herpesvirus 4 (EHV-4) are major viral pathogens of horses that contribute to respiratory disease, abortion, and neurological disorders, leading to economic losses in the equine industry. Accurate serological diagnosis is critical for disease surveillance and control. This study aimed to develop and validate an in-house enzyme-linked immunosorbent assay (ELISA) for detecting antibodies against EHV-1 and EHV-4 in horse sera. Serum samples previously confirmed by virus neutralization test (VNT) and a commercial ELISA were used to optimize the assay. Cut-off values were established using negative controls, and 155 serum samples were tested. The developed ELISA demonstrated 80.64% positivity for EHV-1 and 79.35% for EHV-4. When compared to VNT, the assay showed 85% specificity and 100% sensitivity. A high similarity (99%) was also observed when compared with the commercial ELISA. The overall seroprevalence was found to be 54.19% for EHV-1 and 75.48% for EHV-4, with an SN₅₀ value of 1/10 for both. The validation results confirmed high repeatability and reliability across plates. These findings suggest that the developed in-house ELISA is a practical, cost-effective, and accurate tool for the serological monitoring of EHV-1 and EHV-4 infections in horse populations. Full article