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Topic:Enzyme-Linked Immunosorbent Assay (ELISA)
Molecular and Serological Survey of Severe Fever with Thrombocytopenia Syndrome Virus in Horses from the Republic of Korea. Severe fever with thrombocytopenia syndrome (SFTS) is an emerging zoonotic tick-borne disease in East Asia caused by the SFTS virus (SFTSV). It is to investigate the presence of SFTSV RNA and antibodies in horses from a slaughterhouse and equestrian centers in the Republic of Korea (ROK). A prevalence study of SFTSV-specific RNA and antibodies was designed from 889 horses in the ROK. Serum samples were collected from horses at a slaughterhouse and equestrian centers from 2018 to 2020. To detect the presence of SFTSV, RNA was extracted from the serum samples, and a nested reverse transcriptio...
Plasma melatonin concentration during the early post-partum period in Thoroughbred mares and their foals. The authors aimed to determine the plasma melatonin concentration in mares and their new-born foals in the early post-partum period. Blood samples were collected from the jugular vein of 53 mare-foal pairs within twelve hours after parturition. Plasma melatonin levels were measured by ELISA. The melatonin concentration, adjusted for the moment of parturition using a generalised linear model, was 34.58 pg mL-1 in mares. It was significantly lower (27.63 pg mL-1) in the new-born foals. However, the melatonin concentration declined differently by the end of the twelve hours, it decreased less in...
Fasciola hepatica infection in horses in three governorates in northern Egypt: prevalence and risk factors. Fasciola hepatica is a trematode parasite that affects wide range of mammalian hosts including horses. There are no epidemiological data on fasciolosis in Egyptian horses. Hence, the present study aimed to evaluate the seroprevalence of F. hepatica in horses using ELISA as well as assess the associated risk factors for fasciolosis infection. A total of 400 sera samples of horses from three Egyptian governorates were examined. The overall seroprevalence rate in horses was 13% and the disease was more prevalent in Qalubia (14.17%) and Gharbia (16.67%) which are located at Nile Delta. The likelih...
Regulatory T lymphocytes and selected cytokines in horses with mild to severe equine asthma and different inflammatory cytology profiles. This study determined Tregs and inflammatory cytokines in BALF and peripheral blood (PB) of adult horses with mild and severe asthma and different BALF cytological inflammation profiles. Horses of diverse breeds with asthma (age range: 2-20 years, n = 24) were divided into groups according to the number of points obtained in a standardized clinical scoring system (mild-moderate equine asthma - MEA, severe equine asthma - SEA) and according to the inflammation type based on cytological finding. Plasma levels of IFN-γ, IL-4, IL-10, IL-17 and MMP-9 in the BALF were determined by ELISA. Tregs ...
Validation of a serum ELISA test for cyathostomin infection in equines. Cyathostomins are ubiquitous equine nematodes. Infection can result in larval cyathostominosis due to mass larval emergence. Although faecal egg count (FEC) tests provide estimates of egg shedding, these correlate poorly with burden and provide no information on mucosal/luminal larvae. Previous studies describe a serum IgG(T)-based ELISA (CT3) that exhibits utility for detection of mucosal/luminal cyathostomins. Here, this ELISA is optimised/validated for commercial application using sera from horses for which burden data were available. Optimisation included addition of total IgG-based calibr...
The serological detection of Bovine papillomavirus’s E5 oncoprotein antibodies in horses. Equine sarcoids (ES) are known globally as the most frequent skin tumour affecting horses. These tumours affect the horse's monetary value, they can affect the horse's welfare and can be difficult and expensive to treat. Bovine papillomavirus (BPV) is considered to be the aetiological agent of this tumour, as BPV 1, 2 and 13 have been detected in ES. This is the only known natural cross species infection by a papillomavirus. The BPV genome can be divided into two coding regions: The early region E which encodes the transforming proteins E5, E6 and E7 as well as the replication and transcriptio...
Botulinum neurotoxin type A does not exert concentration-dependent effects on equine articular cartilage in vitro. To determine whether Botulinum neurotoxin type A (BoNT-A) ameliorates the effects of interleukin 1 (IL-1) on equine articular cartilage, or exerts negative effects on normal equine articular cartilage homeostasis in vitro. Methods: Articular cartilage explants from 6 healthy femoropatellar joints of 3 adult horses. Methods: Explants were allocated to the IL-1 challenged or unchallenged group, then exposed to 1 of 6 concentrations of BoNT-A (0, 1, 10, 50, 100, or 500 pg/mL) for 96 hours. To assess BoNT-A's effects on inflammation, prostaglandin E2 (PGE2) was measured in media via ELISA. Matrix ...
Exposure levels of animal allergens, endotoxin, and β-(1,3)-glucan on a university campus of veterinary medicine. The study aimed to determine the allergen, endotoxin and β-(1,3)-glucan concentrations at various areas on a university campus of veterinary medicine. Methods: Dust samples were collected four times a year for three years using electrostatic dust collectors (EDC) at 25 different locations on a campus of veterinary medicine and in laboratories of inorganic chemistry as a control area representing animal-free environment. Major animal allergens from dog, cat, horse, cattle and mouse, domestic mite (DM) allergens, and β-(1,3)-glucan were measured using enzyme immunoassays and endotoxin using th...
Screening and confirmation of recombinant human follistatin in equine plasma for doping control purposes. Recombinant human follistatin (rhFST) is a potential performance-enhancing agent owing to its stimulating effect on muscle growth. Administration of rhFST to athletes is prohibited in human sports by the World Anti-Doping Agency (WADA) and in horseracing according to Article 6 of the International Agreement on Breeding, Racing and Wagering published by the International Federation of Horseracing Authorities (IFHA). For effective control of the potential misuse of rhFST in flat racing, methods for screening and confirmatory analysis are required. This paper describes the development and validat...
Evaluation of the effects of medium-term (57-day) omeprazole administration and of omeprazole discontinuation on serum gastrin and serum chromogranin A concentrations in the horse. Rebound gastric hyperacidity (RGH) secondary to hypergastrinemia has been suggested to contribute to the rapid recurrence of equine squamous gastric disease (ESGD) in horses after discontinuation of omeprazole. Objective: To evaluate changes in serum gastrin and chromogranin A (CgA) concentrations in response to medium-term (57-day) omeprazole treatment and after omeprazole discontinuation. Methods: Fourteen mature Thoroughbred racehorses in simulated race training. Methods: Horses received 2.28 g of oral omeprazole PO q24h for 57 days within a 61-day period, excluding a withholding period...
Antibodies against Borrelia burgdorferi Sensu Lato in Clinically Healthy and Sick Horses: First Report from the Czech Republic. Lyme disease, caused by some strains of bacterial spirochetes Borrelia burgdorferi sensu lato (Bbsl), affects humans but also domestic animals including horses. The primary pathogens in horses in Europe are B. afzelii, B. garinii and B. burgdorferi sensu stricto. To our knowledge, there are no data available on the seropositivity of B. burgdorferi s.l. in horses from the Czech Republic. In this country, horses are mainly used for sport, breeding, and recreational riding in areas where vectors of B. burgdorferi s.l. are present, which is why they are frequently at risk of infection. The aim of ...
Serological and molecular surveillance of West Nile virus in domesticated mammals of peninsular Malaysia. West Nile virus is a mosquito-borne neurotropic pathogen with a wide host range that constitutes a significant risk to public and animal health. There is limited information on WNV infection in domesticated mammals in Malaysia; however, current reports indicate infections in birds, macaques, bats and pigs from Malaysia. In this study, 203 serum samples from cattle, goats, and horses were tested for the presence of anti-WNV IgG using a competitive enzyme-linked immunosorbent assay (c-ELISA). Additionally, using one-step RT-PCR, nasopharyngeal swabs were analyzed for WNV RNA from all 203 animals...
Serological and Bacteriological Surveillance of Glanders Among Horses in Central Region of Iran. Glanders is the oldest and very contagious disease among horses caused by Burkholderia mallei. The disease occurs as a chronic form in horses. Hence, because of the prolonged shedding, numerous horses can potentially get infected by one horse with glanders. Glanders is endemic in Iran and this causes occasional occurrence in horse population of the country. The aim of this study was to determine the incidence of B.mallei infection in horses in two central provinces of Iran. A total of 517 serum samples were collected from stable horses in Tehran and Alborz provinces. Among the studied horses, ...
Validation of an ELISA kit to measure allopregnanolone in human and equine hair. In humans, allopregnanolone plays important roles in a number of different neurodegenerative disorders, and it has been proposed for use in some therapies. Horses are commonly used as animal models for human neurodegenerative diseases, mental and behavioral disorders, and neuropsychiatric disorders, and there is interest in using hair as a biological sample to study hormones in these conditions. We validated the use of a commercial ELISA kit (DetectX allopregnanolone kit; Arbor Assays), which was designed for serum, plasma, feces, urine, and tissue samples, to assess allopregnanolone in hair s...
Serum nerve growth factor in horses with osteoarthritis-associated lameness. Nerve growth factor (NGF) is a neurotrophin that is increased in osteoarthritic joints of horses. In humans, NGF has been associated with pain, and both synovial and serum NGF concentrations are increased in osteoarthritic patients. Studies in humans also have shown that serum NGF concentration can increase with stress. Serum NGF concentration should be evaluated in horses with osteoarthritis-associated lameness. Objective: Quantify and compare serum NGF concentration in horses with osteoarthritis-associated lameness and sound horses. Additionally, the impact of short-term stress on serum NGF ...
Development of a promising antigenic cocktail for the global detection of Babesia caballi in horse by ELISA. In this study, we designed novel truncated Babesia caballi (B. caballi) recombinant proteins from the previously used B. caballi proteins; 134-Kilodalton Protein (rBC134) and Merozoite Rhoptry 48 Protein (rBC48). Then, we evaluated the diagnostic performance of the newly designed proteins when used as a single antigen or when used as cocktail antigen consists of rBC134 full length (rBC134f) + newly designed rBC48 (rBC48t) or newly designed rBC134 (rBC134t) + rBC48t for the detection of B. caballi infection in horse using indirect enzyme-linked immunosorbent assay (iELISA). We used one dose and...
Development and evaluation of a blocking ELISA for serological diagnosis of equine infectious anemia. Equine infectious anemia (EIA) is an important viral disease characterized by persistent infection in equids worldwide. Most EIA cases are life-long virus carriers with low antibody reactions and without the appearance of clinical symptoms. A serological test with high sensitivity and specificity is required to detect inapparent infection. In this study, a B-cell common epitope-based blocking ELISA (bELISA) was developed using a monoclonal antibody together with the EIAV p26 protein labelled with HRP. The test has been evaluated against the standard and with field serum samples globally. This ...
Plasma UCHL-1 as a Biomarker of Brain Injury in Hospitalized Foals With Neonatal Encephalopathy. A plasma biomarker such as ubiquitin carboxyl-terminal hydrolase L1 (UCHL-1) to distinguish neonatal encephalopathy (NE) from other disorders and provide prognostic information would be useful for equine practitioners. In this prospective study, plasma UCHL-1 was measured in 331 hospitalized foals ≤4 days of age. Clinical diagnoses of neonatal encephalopathy only (NE group, n = 77), sepsis only (Sepsis group, n = 34), concurrent sepsis and NE (NE+Sepsis group, n = 85), or neither sepsis nor NE (Other group, n = 101) were made by the attending veterinarian. Plasma UCHL-1 concentration...
Variation in salivary cortisol responses in yearling Thoroughbred racehorses during their first year of training. Thoroughbred horses are bred for competitive racing and undergo intense training regimes. The maintenance of physical soundness and desirable behavioural characteristics are critical to the longevity of a racing career. Horses intended for Flat racing generally enter training as yearlings and undergo introductory training prior to exercise conditioning for racing. This period requires rapid adjustment to a novel environment. As a prey animal, a horse's 'fight-or-flight' response is highly adapted, in which a well-understood component of this response, the hypothalamic-pituitary-axis, is activa...
Relationship between plasma dopamine concentration and temperament in horses. Dopamine (DA) is a neurotransmitter associated with animal behaviors. Along with other neurotransmitters such as oxytocin (OXT) and serotonin (5-HT), DA is also involved in determining the temperament of animals. However, the involvement of DA in horse temperament has not been well elucidated. Therefore, in this study, we aimed to determine the correlation between plasma DA concentration and OXT and 5-HT concentrations and behavioral temperament (eg, docility and friendliness, fearfulness, dominance, and trainability) of horses. Blood samples were collected from 31 horses and the concentration...
Seroprevalence and Risk Factors for Toxoplasma gondii Infection in Horses. is classified as intracellular protozoa and is one of the major zoonotic parasites. Most warm-blooded intermediate hosts, including humans, are commonly infected by this parasite. The epidemiology of infection in Egyptian horses is currently poorly understood. Methods: 420 blood samples were randomly collected from horses raised in four governorates in Northern Egypt (110 each from Giza and Kafr El Sheikh, and 100 each from Qalyubia and Gharbia) to investigate the existence of antibodies against using a commercial ELISA kit, and to ascertain the risk factors for the infection. Results: the ...
Characterization of serum and tissue oxytocinase and tissue oxytocin in the pregnant and non-pregnant mare. Oxytocin is a hormone with functions in: reproduction, maternal bonding, milk ejection, and feeding/social behavior, and is reported to be present in a variety of tissues. Our goal is to characterize oxytocin and leucyl and cystinyl aminopeptidase (LNPEP/oxytocinase), a key regulator of oxytocin in mares. We measured serum and tissue LNPEP by ELISA from ovulation (D0) until D21-22 in non-pregnant (n = 5) and pregnant mares (n = 6); and in periparturient and postpartum mares (n = 18). Placenta (n = 7) and homogenized tissue of diestrus mares (n = 6) were evaluated using prot...
Interleukin-1β and methylprednisolone acetate demonstrate differential effects on equine deep digital flexor tendon and navicular bone fibrocartilage cells in vitro. To investigate the effects of interleukin-1β (IL-1β) and methylprednisolone acetate (MPA) on equine intrabursal deep digital flexor tendon (DDFT) and navicular bone fibrocartilage (NBF) cells in vitro. Methods: Third passage DDFT and NBF cells from 5 healthy donor horses ages 11-17 years euthanized for reasons unrelated to musculoskeletal conditions. Methods: Aggregate cultures were incubated with culture medium alone (control), 10 ng/mL IL-1β, 10 ng/mL IL-1β + 0.05 mg/mL MPA, or 10 ng/mL IL-1β + 0.5 mg/mL MPA for 24 hours. Extracellular matrix (ECM) gene expressions were assessed via rea...
Strangles in equines: An overview. Strangles, caused by Streptococcus equi subspecies equi, is a highly infectious respiratory disease affecting horses and other equines. The disease is economically important and compromises the productivity of equine farm significantly. The disease is characterized by pyrexia, mucopurulent nasal discharge, and abscess formation in the lymph nodes of the head and neck of horses. The disease transmission occurs either directly by coming in contact with infectious exudates or indirectly via fomite transmission. Besides this, carrier animals are the primary and most problematic source of disease i...
Hypoxia signaling in the equine small intestine: Expression and distribution of hypoxia inducible factors during experimental ischemia. Hypoxia inducible factors (HIF) are widely researched in human medicine for their role in different disease processes. The aim of this study was to investigate the expression and distribution of HIF in experimental small intestinal ischemia in the horse. Unassigned: In 14 horses under general anesthesia, segmental jejunal ischemia with 90% reduction in blood flow was induced. The horses were randomly divided into two groups of seven horses, one subjected to ischemic postconditioning (IPoC) by delayed reperfusion, and a control group (group C) undergoing undelayed reperfusion. Intestinal sample...
T Helper 17-Like Regulatory T Cells in Equine Synovial Fluid Are Associated With Disease Severity of Naturally Occurring Posttraumatic Osteoarthritis. Infiltration of cluster of differentiation (CD) 3 (CD3) T cells into the synovium and synovial fluid occurs in most patients with posttraumatic osteoarthritis. During disease progression, proinflammatory T helper 17 cells and anti-inflammatory regulatory T cells infiltrate the joint in response to inflammation. This study aimed to characterize the dynamics of regulatory T and T helper 17 cell populations in synovial fluid from equine clinical patients with posttraumatic osteoarthritis to determine whether phenotype and function are associated with potential immunotherapeutic targets. An imbala...
Evaluation of a mimotope of the Rickettsia outer membrane protein A (OmpA) as an antigen in enzyme-linked immunosorbent assay to detect rickettsiosis in capybaras (Hydrochoerus hydrochaeris), horses (Equus caballus), and opossums (Didelphis sp.). Rickettsia rickettsii is the etiological agent of Rocky Mountain spotted fever, which is an important tick-borne zoonosis and, in Brazil, it causes Brazilian spotted fever, which has high lethality rate. This study aimed to evaluate a synthetic peptide corresponding to a segment of the outer membrane protein A (OmpA) as an antigen in a serological test for the diagnosis of rickettsial infections. The amino acid sequence of the peptide was selected by predicting B cell epitopes using B Cell Epitope Prediction (Immune Epitope Database and Analysis Resource) and Epitopia and OmpA sequences of Ric...
Comparison of Seroprevalence and Identification of Risk Factors for Theileria equi in Horses From Vector-Free and Infested Areas in Southern Brazil. The apicomplexan hemoprotozoan Theileria equi is a tick-borne pathogen that causes disease in equids, and together with Babesia caballi, causes equine piroplasmosis (EP). Many ticks are associated with EP around the world, and in South America three species may be related: Dermacentor nitens, Amblyomma sculptum, and Rhipicephalus microplus, as they are commonly found in horses. Among the species cited above, only R. microplus is found in Rio Grande do Sul state, Southern Brazil. In addition, this state has the only area legally free of R. microplus in Brazil. This study aimed to compare the se...
Acute phase proteins levels in horses, after a single carbohydrate overload, associated with cecal alkalinization. Horses submitted to carbohydrate overload can develop laminitis due to changes in cecal pH and microbiota, followed by an increase in transmural absorption of luminal content, including bacterial toxins. In response to acute injury there is hepatic overproduction of several proteins known as acute phase proteins (APP). Few studies have evaluated protein fractionation to characterize the inflammatory response in acute laminitis. The aim of this study was to test the viability of an experimental model to induce acute laminitis, using a single carbohydrate overload, and the influence of a bufferi...
The effects of cannabidiol on immune function and health parameters in senior horses. Cannabidiol (CBD) has potential to reduce pain and inflammation in humans leading to the interest of use in equine. The purpose of this study was to determine the effects of CBD on immune function by measuring inflammatory cytokines and antibody responses to vaccination, as well as other health parameters in senior horses. Horses were orally-dosed with CBD (2 mg/kg: 13 horses) or control (soy oil: 14 horses) daily for 90 days, from July 2021 to November 2021. Peripheral blood samples were collected on days 0, 30, 60, and 90 before administering treatments. On day 90 all horses were kept on tr...
