Enzyme-Linked Immunosorbent Assay (ELISA) is a widely used analytical technique in equine research for detecting and quantifying specific proteins, hormones, and antibodies in horse biological samples. This method relies on antigen-antibody interactions and employs enzyme-linked antibodies to produce a measurable signal, typically a color change, indicating the presence and concentration of the target molecule. ELISA is applicable in various areas of equine health, including the diagnosis of infectious diseases, monitoring of immune responses, and assessment of physiological conditions. It is valued for its specificity, sensitivity, and ability to process multiple samples simultaneously. This page compiles peer-reviewed research studies and scholarly articles that explore the applications, methodologies, and advancements of ELISA in equine science.
Lee G, Yoon M.Oxytocin (OXT) and vasopressin (AVP) have been associated with social interaction and affiliative behavior in animals. Additionally, AVP is known to affect socially aggressive behavior. In addition, serotonin has an association with aggressive behaviors. The objectives of this study were (1) to evaluate OXT, AVP, and serotonin concentrations in the plasma of horses of different breeds, sexes, and ages and (2) to determine if the neurotransmitters are associated with horse docility and friendliness. This study was performed at Sangju International Equestrian Center. Blood samples were collected...
Quigley A, Sekiya M, Garcia-Campos A, Paz-Silva A, Howell A, Williams DJL, Mulcahy G.Fasciola hepatica is a common parasite of livestock in Ireland, causing significant economic losses and affecting animal welfare. A previous abattoir study of 200 horses led to an estimated 9.5 % prevalence of infection in horses slaughtered in Ireland. However, the epidemiology and pathogenic significance of this infection in this species is not well-described. The objectives of this study were to determine the susceptibility of horses to oral challenge infection with F. hepatica metacercariae, and to document the course of the infection along with serological and biochemical response. We att...
Riley CB, Jenvey CJ, Baker FJ, Corripio A. To determine if an ELISA for measurement of IgA in equine serum could be used to measure concentrations of IgA in foal faeces and to determine correlations with concentrations in the milk of the dam. Faeces from 20 Welsh Cob and Welsh Pony foals and milk from their dams were collected within 12 hours (Day 0) and at 6 days after parturition (Day 6). On Day 6, faeces could not be collected from 2/20 foals, and milk samples could not be collected from 3/20 mares. An equine IgA ELISA validated for serum and plasma was used to measure concentrations of IgA in all samples in triplicate. The preci...
Smith R, Önnerfjord P, Holmgren K, di Grado S, Dudhia J.The diagnosis of tendon injury relies on clinical signs and diagnostic imaging but imaging is subjective and does not always correlate with clinical signs. A molecular marker would potentially offer a sensitive and specific diagnostic tool that could also provide objective assessment of healing for the comparison of different treatments. Cartilage Oligomeric Matrix Protein (COMP) has been used as a molecular marker for osteoarthritis in humans and horses but assays for the protein in tendon sheath synovial fluids have shown overlap between horses affected by tendinopathy and controls. We hypot...
Adamu SG, Hassan M, Ardo MB.A cross-sectional survey was conducted to determine the seroprevalence and risk factors influencing the presence of Brucella spp. antibodies in donkeys in Yobe south senatorial zone, Nigeria. The study was aimed at determining the importance of Brucella spp. infection in donkeys (Equus asinus). A total of 200 sera samples from of 105 males and 95 female donkeys were collected and screened for brucellosis using the rose bengal plate test (RBPT) and the indirect enzyme-linked immunosorbent assay (iELISA). Data obtained were analyzed to determine associations and risk factors. The analysis reveal...
Braucke AFGV, Frederiksen NL, Berg LC, Aarsvold S, Müller FC, Boesen MP, Lindegaard C.Joint pain and osteoarthritis (OA) are some of the most common causes of lameness in horses, and most of the available treatments focus on symptomatic relief without a disease-modifying effect. TRPV1 is a potential target for treating joint diseases, including OA, and the present study aims to investigate if the TRPV1 receptor is present in equine articular tissue and determine whether the number of receptors is upregulated in joint inflammation. Metacarpo/metatarsophalangeal (MCP/MTP) joints from 15 horses euthanised for reasons unrelated to this study were included. Based on synovial fluid a...
Tzelos T, Geyer KK, Mitchell MC, McWilliam HEG, Kharchenko VO, Burgess STG, Matthews JB.Cyathostomins are ubiquitous parasitic nematodes of horses. These worms spend substantial periods as intestinal wall stage encysted larvae, which can comprise up to 90% of the total burden. Several million larvae have been reported in individuals. Emergence of these larvae from the gut wall can lead to life-threatening colitis. Faecal egg count tests, increasingly used by horse owners to inform anthelmintic treatments, do not correlate with the intra-host burden of cyathostomins; this represents a key gap in the diagnostic toolbox. Previously, a cyathostomin Gut Associated Larval Antigen was i...
Wernery U, Joseph S, Raghavan R, Dyer B, Spendrup S.Our investigation has shown that multiple vaccinations with inactivated African horse sickness (AHS) vaccines containing all 9 serotypes and produced at the Central Veterinary Research Laboratory in Dubai, UAE, protect horses from AHS. However, the immunization did not prevent African horse sickness fever (AHSF) in approximately 10% of the vaccinated horses despite high enzyme-linked immunosorbent assay and virus neutralizing antibodies. African horse sickness fever is a very mild form of AHS with similar clinical signs. From all 6 horses which had developed AHSF, no virus was isolated from ED...
Bruschetta G, D'Ascola A, Medica P, Ferlazzo AM.Serotonin (5-hydroxytryptamine [5-HT]) may induce metabolic effects in different cell types, including leukocytes. In horses, 5-HT is involved in physiological and behavioral functions. Physical exercise is known to increase the amounts of 5-HT both in brain and periphery, but so far, the signal mechanism in response to exercise is not known. The aim of the study was to investigate the effect of a racehorse intensive training session on plasma 5-HT levels, serotonin transporter (SERT), 5HT, 5-HT, 5-HT, 5-HT receptor, interleukin-1 beta, and tumor necrosis factor-alpha expression in horse perip...
Assaid N, Mousson L, Moutailler S, Arich S, Akarid K, Monier M, Beck C, Lecollinet S, Failloux AB, Sarih M.West Nile virus (WNV) is one of the most widely distributed mosquito-borne viruses in the world. In North Africa, it causes human cases of meningoencephalitis with fatalities in Algeria and in Tunisia, whereas only horses were affected in Morocco. The aims of this study were to detect WNV in mosquitoes and to determine seroprevalence of WNV in Moroccan horses by the detection of IgG antibodies. A total of 1455 mosquitoes belonging to four different species were grouped by collection site, date, and sex with 10 specimens per pool and tested for 38 arboviruses using a high-throughput chip based ...
Peal BT, Gagliardi R, Su J, Fortier LA, Delco ML, Nixon AJ, Reesink HL.The objectives of this study were to evaluate temporal changes in lubricin, hyaluronan (HA), and HA molecular weight (MW) distributions in three distinct models of equine joint injury affecting the carpal (wrist), tarsal (ankle), and femoropatellar (knee) joints. To establish ranges for lubricin, HA, and HA MW distributions across multiple joints, we first evaluated clinically healthy, high-motion equine joints. Synovial fluid was collected from high-motion joints in horses without clinical signs of joint disease (n = 11 horses, 102 joints) and from research horses undergoing carpal osteo...
Elata A, Mossaad E, Satti R, Matar N, Ohari Y, Xuan X, Inoue N, Suganuma K.In Sudan, donkeys are important animals, providing transportation and income possibilities. However, the prevalence of parasites in donkeys in Sudan has not been thoroughly characterized. Accordingly, in this study, we aimed to detect selected hemoprotozoan parasites in donkeys in West Omdurman, Khartoum State, Sudan, wherein people depend mainly on donkeys for their daily life. In total, 198 blood samples collected from donkeys in a local market in West Omdurman, were screened using serological and molecular diagnostic techniques. Serologically, 52 (26.3%), 56 (28.3%), and 19 (9.6%) samples w...
Ouslimani SF, Tennah S, Azzag N, Derdour SY, China B, Ghalmi F.The aim of this study was to assess the seroprevalence of the in horses in different parts of Algeria and to determine risk factors for the infection. Methods: A total of 736 blood samples were collected from horses of various breeds, gender, coat colors, and ages. All horses came from various farms, racecourses, and equestrian centers. The seroprevalence was investigated by three different methods: Indirect fluorescent antibody test (IFAT) as reference method, enzyme-linked immunosorbent assay (ELISA), and latex agglutination test (LAT). Results: Out of the 736 sera, 178 (24.18%) were positi...
Bannai H, Nemoto M, Tsujimura K, Ohta M.An enzyme-linked immunosorbent assay (ELISA) using a synthetic peptide for the E2 glycoprotein was developed for the serodiagnosis of Getah virus infection in horses. To identify an immunogenic epitope, a series of 20-mer peptides (n = 22) for the E2 protein was screened with pooled sera from horses infected with Getah virus. Peptide P11 (PTEEEIDMHTPPDIPDITLL) showed the strongest reaction. ELISA using P11 (E2-P11-ELISA) detected increased antibody levels in all seven experimentally infected horses and in five out of nine vaccinated horses. Out of 28 naturally infected horses, 25 were sero...
Jones B, Taylor K, Lucas RM, Merritt T, Chicken C, Heller J, Carrick J, Givney R, Durrheim DN.This report describes the challenges encountered in using serological methods to study the historical transmission risk of from horses to humans. In 2017, serology and risk factor questionnaire data from a group of individuals, whose occupations involved close contact with horses, were collected to assess the seroprevalence of antibodies to and identify risk factors associated with previous exposure. 147 participants were enrolled in the study, provided blood samples, and completed a questionnaire. On ELISA testing, antibodies to the Chlamydia genus were detected in samples from 17 participa...
Hannan MA, Murata K, Takeuchi S, Haneda S, Cheong SH, Nambo Y.The objectives of this study were to determine the plasma profile of equine chorionic gonadotropin (eCG) and its association with the formation of supplementary corpus luteum (CL) and plasma progesterone concentrations in embryo transfer Hokkaido native pony recipient mares. Blood samples and transrectal ultrasound examination of the reproductive tract were carried out weekly from the day of ovulation until week 32 of gestation (n = 4). Plasma concentrations of eCG and progesterone were measured by enzyme immunoassays. The eCG concentration was first detectable at week 5 for 2 mares and at wee...
Rui P, Zhao F, Yan S, Wang C, Fu Q, Hao J, Zhou X, Zhong H, Tang M, Hui W, Li W, Shi D, Ma Z, Song T.Hepatitis E virus (HEV) is the causative agent of acute self-limiting hepatitis in humans in developing countries. Hepatitis E virus RNA was first detected in donkeys in Spain, but little is known about the possible presence of HEV in donkeys in China. Objective: To investigate the prevalence of HEV in donkeys in northern China. Methods: Investigation of the prevalence of HEV in donkeys using serological, molecular and phylogenetic approaches. Methods: A total of 401 donkey serum specimens were tested for serological and molecular detection of HEV via enzyme-linked immunosorbent assay and quan...
Zhao S, Smits C, Schuurman N, Barnum S, Pusterla N, Kuppeveld FV, Bosch BJ, Maanen KV, Egberink H.Equine coronavirus (ECoV) is considered to be involved in enteric diseases in foals. Recently, several outbreaks of ECoV infection have also been reported in adult horses from the USA, France and Japan. Epidemiological studies of ECoV infection are still limited, and the seroprevalence of ECoV infection in Europe is unknown. In this study, an indirect enzyme-linked immunosorbent assay (ELISA) method utilizing ECoV spike S1 protein was developed in two formats, and further validated by analyzing 27 paired serum samples (acute and convalescent sera) from horses involved in an ECoV outbreak and 1...
Camino E, Pozo P, Dorrego A, Carvajal KA, Buendia A, Gonzalez S, de Juan L, Dominguez L, Cruz-Lopez F.Serological analysis of equine piroplasmosis (EP), caused by Theileria equi and Babesia caballi, is included in the export testing requirements for most of the countries worldwide, thus involving a high economic impact on equine industry of EP-endemic countries, such as Spain. A total of 3368 serum samples from healthy horses collected prior to export between 2015 and 2018 in Spain were tested for antibodies against T. equi and B. caballi by using a competitive inhibition enzyme-linked immunosorbent assay (cELISA). The overall seroprevalence results in Spain revealed that almost a quarter of t...
Medica P, Giunta RP, Bruschetta G, Ferlazzo AM.Exercise represents a physical stress that challenges homeostasis affecting central and peripheral serotoninergic systems. The influence of the exercise on circulating serotonin (5-hydroxytryptamine [5-HT]) levels depends on training state as well as the exercise protocol. The purpose of the present research was to determine changes of plasma 5-HT in sport horses in response to training (T) and simulated race (SR) and in addition to assess the possible presence of significant differences on circulating 5-HT between two different sessions of exercise. In particular, the research was carried out...
Elzinga SE, Betancourt A, Stewart JC, Altman MH, Barker VD, Muholland M, Bailey S, Brennan KM, Adams AA.Much of the equine population is obese and therefore predisposed to the development of additional health concerns such as equine metabolic syndrome (EMS). However, pharmacologic treatments for EMS are limited. Omega-3 fatty acid supplementation is a therapeutic strategy in humans with metabolic dysfunction that improves insulin sensitivity and reduces inflammation, but the effects of omega-3 fatty acid supplementation in horses with EMS are unclear. Therefore, in this pilot study, 10 mixed-sex and mixed-breed horses with EMS were fed a docosahexaenoic acid (DHA)-rich microalgae containing 16Â ...
Back H, Weld J, Walsh C, Cullinane A.The aim of this study was to identify respiratory viruses circulating amongst elite racehorses in a training yard by serological testing of serial samples and to determine their impact on health status and ability to race. A six-month longitudinal study was conducted in 30 Thoroughbred racehorses (21 two-year-olds, five three-year-olds and four four-year-olds) during the Flat racing season. Sera were tested for the presence of antibodies against equine herpesvirus 1 and 4 (EHV-1 and EHV-4) and equine rhinitis viruses A and B (ERAV and ERBV) by complement fixation (CF) and equine arteritis viru...
Camino E, Dorrego A, Carvajal KA, Buendia-Andres A, de Juan L, Dominguez L, Cruz-Lopez F.Equine piroplasmosis (EP) is a tick-borne protozoan disease caused by Theileria equi and/or Babesia caballi. Clinical signs (fever, pale mucosal membranes, jaundice), anemia and hyperbilirubinemia have been associated with the disease. EP is widespread, has a significant economic impact on the equine industry and remains endemic in Spain. This study was carried out with samples belonging to 140 horses residing in Spain and showing common clinical signs of EP. A blood smear microscopic examination and a comparison between the different results obtained by competitive Enzyme-Linked Immunosorbent...
Di Rubbo A, McNabb L, Klein R, White JR, Colling A, Dimitrov DS, Broder CC, Middleton D, Lunt RA.Maintenance of Hendra virus (HeV) in pteropid bat populations has been associated with spillover events in horses, humans and dogs. Experimental studies have demonstrated infections for several other species including guinea pigs, cats and ferrets. The criteria of a sensitive and specific serological test that is effective for a range of species, but which does not require use of live virus, has not been satisfactorily addressed by currently available tests. We have evaluated the use of two HeV neutralizing monoclonal antibodies (mAbs) in a blocking format enzyme-linked immunosorbent assay (bE...
Vianna AM, Santos AC, Cunha RC, de Souza Stori de Lara AP, Weege GB, Leivas Leite FP.Equine theileriosis is a severe equine disease caused by the protozoan Theileria equi, which is prevalent in tropical and subtropical areas. In this study, a recombinant equi merozoite antigen-2 (rEMA-2) of T. equi was used as an immunogen. Two groups of 10 mice each were divided into control and vaccinated groups. Sixty mares seronegative for theileriosis were divided in two groups, one vaccinated and another group as a control animal. Mice and mares of the vaccinated groups were inoculated with 150 μL of the vaccine containing 50 μg of rEMA-2 and 2 mL of the vaccine containing 200 ...
Witkowska-PiÅ‚aszewicz O, BÄ…ska P, Czopowicz M, Å»migrodzka M, Szarska E, Szczepaniak J, Nowak Z, Winnicka A, CywiÅ„ska A.Development of an anti-inflammatory state during physical training has been postulated in both human and equine athletes, but it is not completely understood. The aim of this study was to investigate whether endurance training changes pro- and anti-inflammatory cytokine profiles within a 20-week training season in young inexperienced endurance horses. Nine Arabian horses were examined in this prospective 20-week follow-up study. Blood samples were analysed 5 times monthly, at rest and after training sessions. Routine haematological examinations were performed. Cytokine patterns including IL-1Î...
Myagmarsuren P, Sivakumar T, Enkhtaivan B, Davaasuren B, Zoljargal M, Narantsatsral S, Davkharbayar B, Mungun-Ochir B, Battur B, Inoue N, Igarashi I....Equine piroplasmosis caused by and is an economically important disease with a worldwide distribution. The objective of the present study was to investigate the seroepidemiology of and in horses reared in various Mongolian provinces. Serum samples prepared from blood collected from horses in 19 Mongolian provinces were screened for antibodies specific to and using enzyme-linked immunosorbent assays based on recombinant forms of merozoite antigen-2 and the 48-kDa merozoite rhoptry protein, respectively. Of 1,282 horses analyzed, 423 (33%) and 182 (14.2%) were sero-positive for and , re...
Minervino AHH, Torres AC, Moreira TR, Vinholte BP, Sampaio BM, Bianchi D, Portela JM, Sarturi C, Marcili A, Barrêto Júnior RA, Gennari SM....The State of Pará has one of the largest herds of equids (horse, donkey and mule) in Brazil, most of these animals are found on cattle farms. Equine theileriosis is a tick-borne disease caused by the parasite Theileria equi and is characterized by fever, anaemia, icterus, intravascular haemolysis, haemoglobinuria, spleen and hepatomegaly, and even death. The present study aimed to determine the prevalence of antibodies against T. equi in equids in the western region of the State of Pará, Brazil, and to identify potential risk factors associated with parasite infection. A cross-sectional stu...
Ye C, Yan W, Xiang H, He H, Yang M, Ijaz M, Useh N, Hsieh CL, McDonough PL, McDonough SP, Mohamed H, Yang Z, Chang YF.Animal leptospirosis is one of the most common zoonotic diseases in the United States and around the world. In a previous study, we applied four recombinant antigens, rLipL21, rLoa22, rLipL32 and rLigACon4-8 of Leptospira interrogans (L. interrogans) for the serological diagnosis of equine leptospirosis (Ye et al, Serodiagnosis of equine leptospirosis by ELISA using four recombinant protein markers, Clin. Vaccine. Immunol. 21:478-483). In this study, the same four recombinant antigens were evaluated for their potential to diagnose canine leptospirosis by ELISA. A total of 305 canine sera that ...
Höglund J, Ljungström BL, Nilsson O, Uggla A.A scolex antigen of the horse tapeworm Anoplocephala perfoliata containing at least 14 different proteins was employed in an enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to A. perfoliata in equine sera. The assay was applied to sera from 426 slaughtered horses with different numbers of worms and with varying degrees of intestinal lesions. As measured by the ELISA, there was a very strong effect on the antibody levels both from the number of tapeworms present and from the intestinal lesion score. However, considerable individual variation was observed between horses wit...
Hoane JS, Yeargan MR, Stamper S, Saville WJ, Morrow JK, Lindsay DS, Howe DK.Neospora hughesi is a recently identified cause of equine protozoal myeloencephalitis. However, the significance of this parasite is poorly understood. An enzyme-linked immunosorbent assay (ELISA) with a recombinant form of the N. hughesi 29-kDa surface antigen (rNhSAG1) was developed for serodiagnosis of equine N. hughesi infections. Parallel ELISA analysis showed that animals immunized or infected with N. hughesi exhibited greater antibody reactivity with rNhSAG1 than with the Neospora caninum homolog, rNcSAG1. The rNhSAG1 ELISA showed 94.4% sensitivity and 95.0% specificity when compared wi...
Traversa D, Fichi G, Campigli M, Rondolotti A, Iorio R, Proudman CJ, Pellegrini D, Perrucci S.Anoplocephala perfoliata (Cestoda, Cyclophyllidea), the commonest intestinal tapeworm of horses, can cause colic, intussusceptions, ileal impactions and intestinal perforations. Common diagnostic techniques for A. perfoliata infection, i.e. coprology and serology, show inherent limitations in terms of sensitivity and specificity and new approaches are thus required. Hence, the present study compared the reliability of coprological, serological (i.e. ELISA) and molecular (i.e. nested PCR) methods in detecting A. perfoliata infection in naturally infected horses and in horses treated with a comb...
Bosler EM, Cohen DP, Schulze TL, Olsen C, Bernard W, Lissman B.By using paired sera the IgM and IgG host responses were analyzed in dogs with ELISA and Western blot techniques. Antibodies in clinical seropositive dogs bound to 4-25 IgM and up to 40 or more IgG antigenic determinants. Early IgM response to the 41-kDa flagellin persisted for at least 9 months and involved as many as seven other peptides. IgG response expanded later in the disease and involved more immunogens than are currently recognized in late human disease. A percentage of asymptomatic dogs that later developed clinical symptoms were seropositive. Immunoblot studies suggested that B. bur...
Hussain MH, Saqib M, Raza F, Muhammad G, Asi MN, Mansoor MK, Saleem M, Jabbar A.Equine piroplasmosis (EP) caused by intraerythrocytic parasites (Theileria equi and Babesia caballi) is an emerging equine disease of world-wide distribution. In Pakistan, the prevalence and incidence of EP are unknown. In order to obtain the first insights into the prevalence of the disease, a total of 430 equids, including 33 mules, 65 horses and 332 donkeys, aging from ≤ 5 to ≥ 10 years of either sex, from five metropolises of Punjab, Pakistan, were serologically tested for the presence of antibodies directed against B. caballi and T. equi, using a competitive enzyme-linked immunosorben...
Magnarelli LA, Anderson JF.Class-specific and polyvalent ELISA were developed to detect IgM antibody or total immunoglobulins to Borrelia burgdorferi in equine sera. Analyses of 122 serum specimens, collected during 1985 from horses and ponies in tick-infested areas of Connecticut, revealed IgM antibody in 41 (34%) samples; titration end points ranged from 1:160 to 1:2,560. In polyvalent ELISA, 73 (16%) of 454 serum specimens contained IgM and/or IgG antibody. Seropositivity was highest (32%) for blood samples collected during May. Both ELISA procedures had comparable sensitivities.
Morgan EE, Miller WH, Wagner B.Skin hypersensitivities (allergies) in horses are often diagnosed using clinical signs only. Intradermal testing or serological assays are diagnostic options to confirm the allergic nature of the disease and to identify the allergen(s). Our objective was to develop an allergen-specific enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody specific for horse IgE and to examine its potential for allergen detection in serum in comparison to intradermal testing. Intradermal testing with 61 allergen extracts was performed on 10 horses affected with skin hypersensitivity. Their sera ...
Liu IK, Bernoco M, Feldman M.Ten fertile feral mares and 6 domestic horses (4 fertile mares, 1 infertile mare, 1 gelding) were immunized with heat-solubilized pig zonae pellucidae by 4 injections equivalent to 2000 or 5000 zonae each at 2-4-week intervals and a booster injection of 20,000 zonae 6-10 months after the last of the initial inoculations. The immune response was reflected by high antibody levels as measured by an enzyme-linked immunosorbent assay (ELISA) using immobilized pig zona antigen. In-vivo inhibition of fertility occurred in 12 (86%) of the 14 fertile mares studied and persisted for a minimum of 7 month...
Ma TW, Li Y, Wang GY, Li XR, Jiang RL, Song XP, Zhang ZH, Bai H, Li X, Gao L.The study aimed to clarify the changes in the concentration of inflammatory mediators, proteases, and cartilage degradation biomarkers in the synovial fluid of joints in an equine osteoarthritis model. Methods: Osteoarthritis was induced in eight Mongolian horses by a sterile intra-articular injection of amphotericin B, which was injected into the left carpal joint in a dose of 2 mL (25 mg/mL). The control group comprised five horses which were injected with an equal dose of sterile physiological saline into the left carpal joint. Synovial fluid was obtained at baseline and every week after in...
Feeney E, Peal BT, Inglis JE, Su J, Nixon AJ, Bonassar LJ, Reesink HL.The objective of this study was to examine temporal variations in synovial fluid composition and lubrication following articular fracture. Post-traumatic osteoarthritis (PTOA) was induced by creating an osteochondral fracture in the middle carpal joint of four horses while the contralateral limb served as a sham-operated control. Horses were exercised on a high-speed treadmill, and synovial fluid was collected pre-operatively and at serial timepoints until 75 days post-operatively. Lubricin and hyaluronic acid (HA) concentrations were measured using sandwich ELISAs, and the molecular weight di...
Halliwell RE, Brim TA, Hines MT, Wolf D, White FH.An enzyme linked immunosorbent assay was developed for the detection of immunoglobulin class specific antibodies to Leptospira interrogans serovar pomona in the serum and aqueous humor of horses. Serum antibody was also assayed by microscopic agglutination tests. Although higher levels of antibody were found in sera from horses with signs of uveitis, the association was not statistically significant. Antibodies to pomona were detected in the aqueous of 12 eyes from the 101 horses sampled at a slaughterhouse, and in most instances, a comparison of the aqueous/serum antibody level with that of t...
Spolidorio MG, Labruna MB, Machado RZ, Moraes-Filho J, Zago AM, Donatele DM, Pinheiro SR, Silveira I, Caliari KM, Yoshinari NH.Blood samples collected from 201 humans, 92 dogs, and 27 horses in the state of Espirito Santo, Brazil, were tested by polymerase chain reaction, indirect immunofluorescence assays, and indirect enzyme-linked immunosorbent assay for tick-borne diseases (rickettsiosis, ehrlichiosis, anaplasmosis, borreliosis, babesiosis). Our results indicated that the surveyed counties are endemic for spotted fever group rickettsiosis because sera from 70 (34.8%) humans, 7 (7.6%) dogs, and 7 (25.9%) horses were reactive to at least one of the six Rickettsia species tested. Although there was evidence of ehrlic...
Wagner B, Childs BA, Erb HN.Skin hypersensitivity is an allergic disease induced in horses by allergens of Culicoides midges. The condition is typically diagnosed by clinical signs and in some horses in combination with allergy testing such as intradermal skin testing or serological allergen-specific IgE determination. Here, we describe an alternative method for allergy testing: a histamine release assay (HRA) that combines the functional aspects of skin testing with the convenience of submitting a blood sample. The assay is based on the principle that crosslinking of allergen-specific IgE bound via high-affinity IgE rec...
Ghazy AA, Shaapan RM, Abdel-Rahman EH.A total of 420 serum samples collected from horses of different ages, sexes and breeds, located at some horse farms in Egypt, were used for serological studies. A crude antigen of the locally isolated Toxoplasma gondii tachyzoites from horse tissues (LA) was used for the detection of T. gondii antibodies in horses. It showed good diagnostic efficiency (38.1%) by Enzyme Linked Immunosorbent Assay (ELISA). To increase this efficiency, an affinity purification process was performed. Two fractions were obtained from LA by CNBr-Sepharose 4B affinity column chromatography named; unbound (LAunb) and ...
Palmer DG, Lyon J, Palmer MA, Forshaw D.To determine the specificity and sensitivity of a commercial copro-antigen ELISA for the detection of Fasciola hepatica infection in cattle and sheep and to assess the suitability of the test for use in horses. Methods: Testing was done on more than 100 negative faecal samples from each of sheep, cattle and horses and on at least 40 positive faecal samples from each species. Positive samples were selected based on a positive sedimentation test for liver fluke eggs. Faecal samples of animals from Western Australia, which is free of liver fluke infection, served as negative controls. Specificity...
Peeters LM, Janssens S, Goddeeris BM, De Keyser K, Wilson AD, Kaufmann C, Schaffartzik A, Marti E, Buys N.Insect bite hypersensitivity (IBH) in horses represents an immunoglobulin E (IgE)-mediated hypersensitivity to salivary antigens from biting midges (Culicoides spp.). The aim of this study was to evaluate and compare the performances of IgE ELISAs using recombinant Culicoides spp. Obsoletus group salivary gland antigens or crude whole body extracts ('ObsWBE'), C. nubeculosus recombinant proteins (Culn1, 3, 4, 5, 7, 8 and 10) and Obsoletus group recombinant proteins (Culo1 and 2). IgE levels were measured in plasma of 343 Warmblood horses classified as IBH-affected (n=167) and IBH-unaffected (n...
Xuan X, Nagai A, Battsetseg B, Fukumoto S, Makala LH, Inoue N, Igarashi I, Mikami T, Fujisaki K.Serum samples from horses in the States of Sao Paulo and Mato Grosso do Sul, Brazil were examined for diagnosis of equine piroplasmosis by both the latex agglutination test (LAT) and enzyme-linked immunosorbent assay (ELISA) with recombinant antigens. Of the 47 samples analyzed, 38 (81%) and 42 (90%) samples were positive for B. equi infection and B. caballi infection, respectively. In addition, 35 (75%) samples were positive for both B. equi and B. caballi infections. These results indicate that equine piroplasmosis is widespread and therefore a cause for serious concern in the States of Sao ...
Duarte CG, Bonilla C, Guimarães G, Machado de Avila RA, Mendes TM, Silva W, Tintaya B, Yarleque A, Chávez-Olórtegui C.In this work, an anti-loxoscelic serum was produced by immunizing horses with a recombinant dermonecrotic protein from Loxosceles intermedia (rLiD1). Anti-rLiD1 antibodies were able to recognize different species of Loxosceles venoms by Western Blot and ELISA. The efficacy of anti-rLiD1 serum against the toxic effects of Loxosceles laeta (Peru) venom was tested, showing that anti-rLiD1 serum can neutralize those effects. This study confirms that recombinant proteins can be good candidates to replace crude venoms for antivenom production.
Lang A, de Vries M, Feineis S, Müller E, Osterrieder N, Damiani AM.A peptide-based enzyme-linked immunosorbent assay (ELISA) for discrimination between serological responses to equine herpesvirus type 1 (EHV-1) and 4 (EHV-4) was developed. Three and four peptides for EHV-1 and EHV-4, respectively, were designed and studied initially in the ELISA using sera from foals infected experimentally. The most promising peptide pair, derived from EHV-1 glycoprotein E and EHV-4 glycoprotein G, was evaluated further using acute and convalescent sera from horses infected experimentally and naturally as well as a panel of horse field sera. Ten pre- and post-vaccination ser...
Gershwin LJ, Netherwood KA, Norris MS, Behrens NE, Shao MX.Vaccination of horses is performed annually or semi-annually with multiple viral antigens, either in a combination vaccine or as separate injections. While this practice undoubtedly prevents infection from such diseases as rabies, equine influenza, West Nile virus, and equine herpes virus, the procedure is not without repercussions. Hypersensitivity reactions, including fatal anaphylactic shock, after vaccination, although uncommon, have increased in incidence in recent years. Studies reported herein document the development of IgE antibodies against non-target antigen components of equine vir...
Shane BS, Issel CJ, Montelaro RC.A sensitive specific enzyme-linked immunosorbent assay utilizing purified p26 antigen was developed for the detection of antibodies to equine infectious anemia virus in naturally and experimentally infected horses. Generally, antibodies to the virus could be detected by the enzyme-linked immunosorbent assay 3 to 4 days earlier than by the standard agar gel immunodiffusion test, and they could be detected more reliably in horses with weak or equivocal agar gel immunodiffusion test reactions. The enzyme-linked immunosorbent assay was also successfully applied to the detection of p26 antigen in t...
Cursino AE, Vilela APP, Franco-Luiz APM, de Oliveira JG, Nogueira MF, Júnior JPA, de Aguiar DM, Kroon EG.Equine infectious anemia (EIA) has a worldwide distribution, and is widespread in Brazil. The Brazilian Pantanal presents with high prevalence comprising equine performance and indirectly the livestock industry, since the horses are used for cattle management. Although EIA is routinely diagnosed by the agar gel immunodiffusion test (AGID), this serological assay has some limitations, so PCR-based detection methods have the potential to overcome these limitations and act as complementary tests to those currently used. Considering the limited number of equine infectious anemia virus (EIAV) seque...
Chong YH, Ball JM, Issel CJ, Montelaro RC, Rushlow KE.Defined segments of the transmembrane envelope glycoprotein (gp45) of equine infectious anemia virus were expressed as TrpLE fusion proteins and examined for their reactivity in Western immunoblots against a diverse panel of equine immune sera. The most immunogenic region of gp45 was localized to its amino terminus, positioned between the hydrophobic fusion and the transmembrane domains. A series of overlapping synthetic peptides were used in enzyme-linked immunosorbent assays to define an immunodominant epitope within this region. In contrast, the carboxy-terminal half of gp45 displayed both ...
Pozio E, Sofronic-Milosavljevic L, Gomez Morales MA, Boireau P, Nöckler K.We assessed a serological method for detecting Trichinella infection in horses, specifically, an ELISA using three antigens to detect anti-Trichinella IgG (i.e. a synthetic tyvelose glycan-BSA (stg-BSA) antigen, an excretory/secretory (ES) antigen, and a crude worm extract (CWE) antigen). Serum samples were collected from 2502 horses (433 live horses from Romania and 2069 horses slaughtered in Italy and originating from Italy, Poland, Romania, and Serbia). Serum samples were also taken from horses experimentally infected with different doses of T. spiralis and T. murrelli larvae, as controls. ...
Cullinane A, Quinlivan M, Nelly M, Patterson H, Kenna R, Garvey M, Gildea S, Lyons P, Flynn M, Galvin P, Neylon M, Jankowska K.In 2006 there was an outbreak of equine infectious anaemia (EIA) in Ireland. This paper describes the use of the diagnosis of clinical and subclinical cases of the disease. In acute cases the ELISAs and the immunoblot were more sensitive than the AGID. In one mare, fluctuating antibody levels were observed in all the serological assays before it seroconverted by AGID. Viral RNA and DNA were detected by RT-PCR and PCR in all the tissues from the infected animals examined postmortem. The PCR detected viral DNA in plasma regardless of the stage of the disease. In contrast, the RT-PCR detected RNA...
Bullone M, de Lagarde M, Vargas A, Lavoie JP.The identification of serum biomarkers of lung inflammation would facilitate the diagnosis of inflammatory airway disease (IAD) in horses. Objective: Horses with IAD have higher serum concentrations of markers of inflammation compared to controls. Methods: Twelve horses with IAD and 10 control horses. Methods: This was a prospective case-control study. Blood and BALF were collected from horses with IAD and controls. Serum concentration of surfactant protein D (SP-D), haptoglobin, serum amyloid A (SAA) and of the soluble form of triggering receptor expressed on myeloid cells 1 (sTREM-1) was mea...
Cook RF, Gann SJ, Mumford JA.An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of serum antibodies to equine arteritis virus (EAV). Results from this assay produced a good correlation with results from virus neutralisation tests in horses which had not been regularly vaccinated with commercially available mammalian tissue culture-derived viral vaccines. Vaccination of some horses with tissue culture-derived vaccines induced the formation of antibodies to bovine serum. These antibodies reacted with the bovine protein contaminants in the EAV ELISA antigen, producing false-positive results. Non-vir...
