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.
Cacciolatti C, Meyer-Ficca ML, Southwood LL, Meyer RG, Bertolotti L, Zarucco L.To evaluate effects of poly(ADP-ribose) polymerase-1 (PARP1) inhibitors on the production of tumor necrosis factor-α (TNF-α) by interferon-γ (IFN-γ)- and lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMCs) of horses as an in vitro model of inflammation in horses. Methods: 1,440 samples of PBMCs from 6 healthy research horses. Methods: From heparinized whole blood samples, PBMC cultures were obtained. An initial dose-response trial on 48 PBMC samples from 2 horses (24 samples each) was used to determine concentrations of IFN-γ and LPS for use as low- and high-lev...
Bannai H, Nemoto M, Tsujimura K, Yamanaka T, Kokado H.Getah virus causes fever, skin eruptions, and limb edema in horses. For a high-throughput and time-saving method for serodiagnosis, we explored immunogenic antigens of Getah virus, and established an enzyme-linked immunosorbent assay (ELISA) using a recombinant protein. Western blot analysis using sera from infected horses showed strong reaction with viral antigens around 46 kDa corresponding to E1 or E2 glycoproteins. Recombinant E2 (rE2) protein reacted more strongly with infected horse sera than did rE1 protein in both Western blotting and ELISA. In ELISA using rE2 protein (rE2-ELISA), fo...
Wang P, Song J, Song R, Zhang M, Wu L, Li F, Yan Y, Zhou J, Chahan B, Liao M.Babesia caballi (Nuttal, 1910) is one of the causative agents of equine piroplasmosis which causes economic losses to horse industry in China. There is an urgent need for rapid detection method for B. caballi infection in Xinjiang Province, China. To prepare monoclonal antibodies (mAbs) against Bc48 gene of B. caballi (Xinjiang local strains) and establish colloidal gold-immunochromatographic (ICT) assay for diagnosis of the disease, recombinant Bc48 was expressed and purified from Escherichia coli. With purified Bc48 as immunogen in mice, three hybridoma cells named 11F4, 1H2 and 7F4 secretin...
Rockstroh A, Moges B, Berneck BS, Sattler T, Revilla-Fernández S, Schmoll F, Pacenti M, Sinigaglia A, Barzon L, Schmidt-Chanasit J, Nowotny N....Tick-borne encephalitis virus (TBEV) and West Nile virus (WNV) are important arthropod-borne zoonotic flaviviruses. Due to the emergence of WNV in TBEV-endemic regions co-circulation of both viruses is increasing. Flaviviruses are structurally highly similar, which leads to cross-reacting antibodies upon infection. Currently available serological assays for TBEV and WNV infections are therefore compromised by false-positive results, especially in IgG measurements. In order to discriminate both infections novel diagnostic methods are needed. We describe an ELISA to measure IgG antibodies specif...
Saini S, Singha H, Siwach P, Tripathi BN.Interleukin (IL)-4 and IL-10 activate plethora of immune cells and induce the humoral immune response. However, recombinant version of horse IL-4 and IL-10 has not been investigated to understand their immunomodulating activities. This study aimed to produce recombinant horse mature IL-4 and IL-10 in . Immune-modulating activities of recombinant horse IL-4 and IL-10 were investigated in peripheral blood mononuclear cells (PBMCs). Methods: Equine PBMCs were stimulated with recombinant IL-4 and IL-10. A proliferation of PBMCs was measured by XTT assay and cytokines induction was measured by enzy...
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...
Blommaert D, Sergeant N, Delehedde M, Jouy N, Mitchell V, Franck T, Donnay I, Lejeune JP, Serteyn D.A-kinase anchor protein 4 (AKAP4) is playing a central role in flagellar structure, chemotaxis, capacitation and sperm motility. In mammals, AKAP4 is expressed during spermatogenesis. AKAP4 is synthesized as a precursor, proAKAP4, which is cleaved into mature AKAP4 during fibrous sheath assembly. The proAKAP4 is a good indicator of sperm quality in humans and boars. The aims of this work were to study the expression, the localization and the concentration of proAKAP4 and AKAP4 in equine semen, and to evaluate the possible correlation between the total and progressive motility and the concentra...
Rissanen K, Galat M, Kovalenko G, Rodnina O, Mikharovskyi G, Must K, Jokelainen P.Horses are recognized as important hosts for the zoonotic parasite Toxoplasma gondii, mainly because meat of infected horses can be a source of human T. gondii infections. However, the baseline knowledge on equine T. gondii infections is limited and lacking from many countries. This seroepidemiological study was set in Ukraine, a country where little is known about T. gondii prevalence in any host species. Methods: We investigated 78 serum samples from horses, collected from Kyiv and Lviv regions of Ukraine, using a commercial enzyme-linked immunosorbent assay (ELISA) and a commercial modified...
Cruz I, Vinhas AR, Dubey JP, Cardoso L, Cotovio M, Lopes AP.Neospora spp. are intracellular protozoa with worldwide distribution and closely related to Toxoplasma gondii, which can infect a variety of mammals including horses. From September 2013 to June 2014, 185 horses from northern, central and southern parts of mainland Portugal were randomly sampled and tested for detection of immunoglobulin (Ig) G antibodies to Neospora spp. using an indirect multi-species enzyme-linked immunosorbent assay (ELISA) commercial test (ID Screen® Neospora caninum Indirect Multi-species; ID.vet Innovative Diagnostics, Grabels, France). Two horses (1.1%; CI: 0.1-3.8%),...
Campos JBV, André MR, Gonçalves LR, Freschi CR, Santos FM, de Oliveira CE, Piranda EM, de Andrade GB, Macedo GC, Machado RZ, Herrera HM.Brazilian Pantanal is the world´s largest wetland ecosystem, where cattle's ranching is the most important economic activity. The objective of this study was to compile some epidemiological features on equine piroplasmids from the Nhecolândia sub-region of Pantanal wetland through the evaluation of the patterns of T. equi and B. caballi infections in different groups of horses; identification of the tick species that infest horses; and to study phylogenetic relationships among Theileria equi 18S rRNA gene sequences. During October 2015, blood and serum samples were collected from 170 horses ...
Ekman S, Lindahl A, Rüetschi U, Jansson A, Björkman K, Abrahamsson-Aurell K, Björnsdóttir S, Löfgren M, Hultén LM, Skiöldebrand E.Molecular serum markers that can identify early reversible osteoarthritis (OA) in horses are lacking. Objective: We studied serum concentrations of a novel cartilage oligomeric matrix protein (COMP) neo-epitope in horses subjected to short-term exercise and with acute lameness. The effects of circadian rhythm and age were also evaluated. Methods: Longitudinal studies in healthy horses and cross-sectional comparison of lame and non-lame horses. Methods: Sera were collected from five horses before and after short-term interval exercise and during full-day box rest. Sera from 32 acutely lame hors...
Bonelli F, Rota A, Aurich C, Ille N, Camillo F, Panzani D, Sgorbini M.Salivary cortisol provides information about free plasma cortisol concentration, and salivary sampling is a noninvasive well-tolerated procedure. The aim of this study was to validate a commercial enzyme immunoassay for the determination of salivary cortisol in donkeys. Saliva samples were collected in four donkey stallions on 13 nonconsecutive days at 8:30 AM to avoid circadian variation. Animals were already accustomed to be handled. Saliva was collected using a swab inserted at the angle of the lips, placed onto the tongue for 1 minute and returned into a polypropylene tube. Tubes were cen...
Radwanski NE, Morris DO, Boston RC, Cerundolo R, Lee KW.The long-term effects of allergen specific immunotherapy (ASIT) on concentrations of circulating immunoglobulin E (IgE) and immunoglobulin G (IgG) in horses have not been reported. Objective: To document changes in clinical severity of horses with atopic dermatitis (AD) and to monitor allergen-specific IgE and IgG concentrations during a two-year course of ASIT. Methods: Nineteen client-owned horses with a conditional diagnosis of AD. Methods: Three ASIT groups were randomly assigned based upon results obtained by either intradermal testing (IDT) for regional allergens (n = 7); enzyme-linked...
Rütten S, Schrödl W, Abraham G.Glucocorticoids are important drugs in the treatment of many inflammatory, autoimmune and allergic diseases in humans and animals. We investigated the effects of hydrocortisone and dexamethasone on TNF-α, IL-1Ra and INF-γ release in stimulated whole blood cell culture from healthy horses. Whole blood cell cultures proved to be useful for the characterization of the anti-inflammatory properties of new drugs. Diluted equine whole blood was exposed to lipopolysaccharide (LPS) and PCPwL (a cocktail consisting of phythemagglutinin E, concanavalin A, pokeweed mitogen and lipopolysaccharide) in the...
Victor S, Binnmyr J, Lampa E, Rask-Andersen A, Elfman L.Horses are an important source of allergens, but the distribution of horse allergens is poorly understood. Five horse allergens have been identified, Equ c 1-4 and 6. Equ c 4 seems to be an important allergen, with an IgE-binding frequency of 77% in horse-sensitized individuals. The aim of this study was to investigate levels of horse allergen Equ c 4 in dander, saliva and urine from ten horse breeds. The study population included 170 horses (87 mares, 27 stallions, 56 geldings) from ten breeds. Horse dander, saliva and urine samples were collected. Levels of horse allergen Equ c 4 were quanti...
Cavalcanti RTC, Teixeira PAC, Levy RS, Pereira HMG, Aquino Neto FR.Erythropoiesis-stimulating agents (ESAs) have been used in horses for doping purposes to increase the performance of these animals in endurance sports. Currently, enzyme-linked immunosorbent assay (ELISA) and mass spectrometry methods are used to detect ESA abuse in equines. However, the sarcosyl polyacrylamide gel-electrophoresis (SAR-PAGE) technique could also be used, since its application in human doping control is well established and has proven to be more sensitive. In this work, the SAR-PAGE method was used to detect recombinant human erythropoietin (rHuEPO), novel erythropoiesis stimul...
Beck C, Leparc-Goffart I, Desoutter D, Debergé E, Bichet H, Lowenski S, Dumarest M, Gonzalez G, Migné C, Vanhomwegen J, Zientara S, Durand B....New Caledonia and French Polynesia are areas in which arboviruses circulate extensively. A large serological survey among horses from New Caledonia and French Polynesia was carried out to investigate the seroprevalence of flaviviruses in the horse population. Here, 293 equine sera samples were screened for flaviviruses using a competitive enzyme-linked immunosorbent assay (cELISA). The positive sera were then confirmed using a flavivirus-specific microsphere immunoassay (MIA) and seroneutralization tests. This serosurvey showed that 16.6% (27/163) and 30.8% (40/130) of horses were positive for...
Kummer LL, Govaere J, Egri B.Twenty-eight warmblood mares were monitored during their late pregnancy in the Teaching Hospital of Ghent University. The reliability of two commercial assays (enzyme immunoassay and glutaraldehyde coagulation test) used for determining the IgG concentrations of their newborn foals was tested. Mammary secretions were examined at the time of foaling (T0), and then 4 (T1) and 8 (T2) hours after foaling by refractometry and electrophoresis. The foals' blood IgG levels were measured at T1 and T2 as a routine clinical diagnostic examination using two different commercial test kits (SNAP Foal Ig and...
Naves JHFF, Oliveira FG, Bicalho JM, Santos PS, Machado-de-Ávila RA, Chavez-Olortegui C, Leite RC, Reis JKP.Equine infectious anemia (EIA) is a disease caused by a Lentivirus that is currently controlled exclusively by identification of seropositive animals. In most countries, including Brazil, the official diagnostic test for EIA is the agar gel immunodiffusion test (AGID). Although this assay has a high specificity it can produce false negative reactions or equivocal results due to weak precipitation lines, especially in samples from donkeys, mules or newly infected equids. In this pioneering study, it was used overlapping synthetic peptide pools to map and identify a consensus, widely recognised ...
Bannai H, Nemoto M, Tsujimura K, Yamanaka T, Kokado H, Kondo T.In order to establish an efficient system for serological diagnosis of equine viral arteritis in Japan, we compared enzyme-linked immunosorbent assays (ELISAs) provided by two manufacturers (Nisseiken Co., Ltd., Tokyo, Japan, and VMRD Inc., Pullman, WA, U.S.A.) by testing a series of horse sera. The results revealed that 159 of 160 virus-neutralizing (VN) antibody-positive serum samples were positive in both the Nisseiken-ELISA and VMRD-ELISA. Of the VN-negative sera (n=157), 134 and 154 samples were negative in the Nisseiken-ELISA and VMRD-ELISA, respectively. Sensitivity was 99.4% for both t...
Verdon M, Lanz S, Rhyner C, Gerber V, Marti E.Genetic, epidemiologic, and clinical evidence suggests that, in horses, there are manifestations of hypersensitivity that can occur together. Objective: To investigate whether concurrent insect bite hypersensitivity (IBH) and severe equine asthma (EA) is associated with higher allergen-specific and total serum immunoglobulin E (IgE) concentrations than only EA or IBH. Methods: Healthy control horses (C, n = 40), horses with IBH (IBH, n = 24), severe EA (EA, n = 18), and both conditions (IBH/EA, n = 23) were included. Methods: In our retrospective comparative study, sera from ho...
Jacobsen S, Berg LC, Tvermose E, Laurberg MB, van Galen G.Neutrophil gelatinase-associated lipocalin (NGAL) has been shown to be a useful marker of kidney injury in people and dogs, but has not been described in horses. Objective: The aim of the study was to validate a commercially available porcine-specific ELISA to measure serum concentrations of equine NGAL. Methods: Intra- and interassay imprecisions were evaluated by multiple measurements on equine serum pools. Assay inaccuracy was determined by the linearity under dilution. Overlapping performance was assessed by measuring NGAL concentrations in horses with normal and elevated serum creatinine ...
Hemida MG, Perera RAPM, Chu DKW, Ko RLW, Alnaeem AA, Peiris M.West Nile virus (WNV) is an important emerging zoonotic arbovirus giving rise to clinical syndromes of varying severity in humans and horses. Culex mosquitoes are the main vector. Although WNV has been reported in many countries in the Middle East and Asia, little is known about its prevalence in equine populations in the Arabian Peninsula. We have carried out a serological study on 200 horses to assess WNV infection in the Eastern and Central regions of Saudi Arabia in 2013-2015. Sera were tested for the presence of WNV antibodies in parallel using a commercial enzyme-linked immunosorbent ass...
Halassy B, Kurtović T, Lang Balija M, Brgles M, Tunjić M, Sviben D.The hyperimmune horse plasma (HHP), prepared through active immunisation of horses with an antigen of interest, is the most common starting material for antitoxin (animal antibody-based therapeutics) production. Precise IgG quantification in plasma is a prerequisite for accurate estimation of the purification process efficiency. Although immunoglobulins from HHP have been purified for over a century, there is still no in vitro method for precise and accurate determination of IgG content in HHP. For this reason, the purification process efficiency has been assessed by antibody activity measurem...
Singh S, Dohre SK, Kamthan A, Pal V, Karothia BS, Singha HS, Kumar S.Glanders is a contagious and highly fatal disease of equines with zoonotic potential. It is caused by a Gram-negative, nonmotile bacterium Burkholderia mallei. Complement fixation test (CFT) is one of the most commonly used tests for diagnosis of glanders; however, it has some limitations. A recombinant-truncated Burkholderia intracellular motility A (BimA) protein-based indirect enzyme-linked immunosorbent assay (iELISA) was previously reported by us for glanders diagnosis, which has been re-optimized in this study using a panel of glanders positive (n = 75) and glanders negative (n = 227...
Flanders JA, Wack RF, Pusterla N, Mapes SM, Collins D, Gamble KC.Infection by equine herpesvirus (EHV) strains (EHV-1, EHV-9) in ursid species, including polar bears ( Ursus maritimus), has been associated with neurological disease and death. A serosurvey of captive exotic equid and polar bear populations in US Association of Zoos and Aquaria institutions was performed to determine the prevalence of EHV strains using quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA) tests. Equid species surveyed included zebra ( Equus spp.), Przewalski's wild horse ( Equus ferus przewalskii), Persian onager ( Equus hemionus), and So...
Schnabel CL, Wemette M, Babasyan S, Freer H, Baldwin C, Wagner B.Chemokines are soluble molecules directing immune cell trafficking and homing, mediating inflammation, and initiating immune responses to infection. In horses, the analysis of chemokines has been limited by the lack of specific antibodies. We generated mAbs specific for the equine C-C motif chemokine ligands (CCL) CCL2 (MCP-1), CCL3 (MIP-1α), CCL5 (RANTES) and CCL11 (eotaxin) using hybridoma technology. Antibody specificity was confirmed by intracellular staining of Chinese Hamster Ovary cells transfected with expression vectors encoding for CCL2, CCL3, CCL5, or CCL11. Transfectants were stai...
Dourine is a parasitic venereal disease of equines caused by T. equiperdum. Humoral antibodies are found in infected animals, but diagnosis of dourine must include history, clinical, and pathological findings in addition to serology. Complement Fixation Test (CFT) is the Office International des Epizooties (OIE) recommended test for international trade; however, some uninfected equines may give inconsistent or nonspecific reactions in CFT due to the anticomplementary effects of their sera. In this study an Indirect Enzyme Linked Immunosorbent Assay (iELISA) was developed. This test could be us...
Hemida MG, Alhammadi M, Daleb A, Alnaeem A.African horse sickness virus (AHSV) is one of the most devastating viral diseases of the family Equidae. Infection with AHSV threatens not only the Saudi equine industry but also the equine industry worldwide. This is due to the high morbidity and mortality rates among the infected population of up to 100%. The World Organisation for Animal Health (OIE) lists AHSV among its notifiable diseases; this requires Member Countries to monitor the situation with regard to AHSV very carefully in order to avoid the spread of the virus. The OIE also suggests the systematic monitoring of AHSV in the equin...
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...
Langner KF, Darpel KE, Drolet BS, Fischer A, Hampel S, Heselhaus JE, Mellor PS, Mertens PP, Leibold W.The objective of this study was to compare and analyze three common diagnostic methods for summer eczema (SE) in horses, an allergic dermatitis caused by bites of Culicoides spp. Nine horses with a medical history of SE and nine control animals were intradermally challenged with whole body extracts (WBE) and the saliva of a native (C. nubeculosus) and exotic (C. sonorensis) Culicoides species. Blood and serum samples of the horses were examined for basophil reactivity by a histamine release test (HRT) and for Culicoides-specific serum immunoglobulin E (IgE) and G (IgG) by enzyme-linked immunos...
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...
Joó K, Bakonyi T, Szenci O, Sárdi S, Ferenczi E, Barna M, Malik P, Hubalek Z, Fehér O, Kutasi O.West Nile virus (WNV) mainly infects birds, horses and humans. Outcomes of the infection range from mild uncharacteristic signs to fatal neurologic disease. The main objectives of the present study were to measure serum IgG and IgM antibodies in naturally exposed and vaccinated horses and to compare results of haemagglutination inhibition test (HIT), enzyme-linked immunosorbent assay (ELISA) and plaque reduction neutralisation test (PRNT). Altogether 224 animals were tested by HIT for WNV antibodies and 41 horses were simultaneously examined by ELISA and PRNT. After primary screening for WNV a...
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...
Ozaki H, Sugiura T, Sugita S, Imagawa H, Kida H.Antibodies to the nonstructural protein (NS1) of A/equine/Miami/1/63 (H3N8) influenza virus were detected exclusively in the sera of mice experimentally infected with A/Aichi/2/68 (H3N2) and horses infected with A/equine/Kentucky/1/81 (H3N8) or A/equine/La Plata/1/93 (H3N8), but not in those of the animals immunized with the inactivated viruses, by enzyme-linked immunosorbent assay (ELISA) using a recombinant NS1 as antigen. The results indicate that the present method is useful for serological diagnosis to distinguish horses infected with equine H3 influenza viruses from those immunized with ...
Bartova E, Machacova T, Sedlak K, Budikova M, Mariani U, Veneziano V.The consumption of horse meat has been epidemiologically linked to clinical toxoplasmosis in humans and neosporosis that may cause clinical illness in horses. Here we determined seroprevalence of antibodies against Toxoplasma gondii Nicolle et Manceaux, 1908 and species of Neospora Dubey, Carpenter, Speer, Topper et Uggla, 1988 in horses from Italy. Blood samples were collected from 643 apparently healthy horses from 60 farms of 51 municipalities in southern Italy. The presence of antibodies against T. gondii and Neospora spp. were detected by indirect fluorescence antibody test (IFAT); a titr...
Bhide M, Yilmaz Z, Golcu E, Torun S, Mikula I.The aim of the study was to determine the seroprevalence of anti-Borrelia burgdorferi antibodies in a population of Turkish dogs and horses, as well as to compare the sensitivity of novel flow-cytometry-based borreliacidal antibody test (BAT) with ELISA assay. Serum samples collected from 400 dogs and 300 horses were tested with enzyme-linked protein A/G assay (ELPAGA), using Borrelia whole cell antigens. ELPAGA test showed 93 dogs (23.2%) and 18 horses (6%) serologically positive for anti-Borrelia antibodies. In parallel testing of sera with BAT, we found 27.75% positive dogs and 6.33% positi...
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...
Stefanciková A, Adaszek Ł, Pet'ko B, Winiarczyk S, Dudinák V.In the course of epizootological research on Lyme borreliosis in animals, the serological evidence of this zoonosis in horses and cattle from different voivodships of Poland was screened. We also discussed some diagnostic problems of Lyme borreliosis resulting from, in addition to other factors, genetic and geographical heterogeneity isolates B. burgdorferi s.l. used as antigens. Using ELISA from 395 sera of horses the total mean seroprevalence for anti-Borrelia IgG antibodies 25.6% was observed. In the respective years, significant differences in the mean seroprevalence were not recorded. In ...
Elschner MC, Melzer F, Singha H, Muhammad S, Gardner I, Neubauer H.Glanders, caused by Burkholderia (B.) mallei is a notifiable zoonotic disease in equidae. For international trade and movement of equids, certificates of negative serological test results for antibodies against B. mallei are required. To date, the complement fixation test (CFT) is the mandatory test to issue these health certificates. The CFT is difficult to standardize and, due to its poor specificity, often leads to false-positive reactions resulting in trade restrictions with considerable financial consequences. In the present study, the new ID Screen Glanders Double Antigen Multispecies EL...
Dordas-Perpinyà M, Yanez-Ortiz I, Sergeant N, Mevel V, Bruyas JF, Catalán J, Delehedde M, Briand-Amirat L, Miró J.ProAKAP4 is the precursor of AKAP4 (A-kinase Anchor protein 4), the main structural protein of the fibrous sheath of sperm. The amount of proAKAP4 reflects the ability of spermatozoa to maintain the flagellum activity and functionality up to the site of fertilization and is positively correlated with progressive motility in several mammalian species. The aim of this study was to investigate the relationship between proAKAP4 concentration with horse sperm motility descriptors and spermatic motile subpopulations. For this purpose, a total of 48 ejaculates from 13 different stallions were analyze...
Proudman CJ, Holdstock NB.A novel serological assay which measures IgG(T) specific for a 12/13 kDa protein of the equine tapeworm Anoplocephala perfoliata was used as part of a colic outbreak investigation. A training/rehabilitation yard for Thoroughbreds and Arabs was found to have an increasing incidence of colic over a 5 year period, culminating in a peak incidence of 1.15 episodes/horse year at risk. Four animals suffered from ileal impaction colic which necessitated surgical management. A case-control study design suggested a strong association between tapeworm infection and colic, with evidence of a dose-response...
Eder C, Curik I, Brem G, Crameri R, Bodo I, Habe F, Lazary S, Sölkner J, Marti E.To investigate whether allergen-specific IgE production is influenced by environmental and genetic factors, IgE levels against 2 mould extracts (Alternaria alternata [Alt a] and Aspergillus fumigatus [Asp f]) and against recombinant (r) rAlt a 1, rAsp f 7 and rAsp f 8 were determined by ELISA in sera from 448 Lipizzan horses living in 6 studfarms. Statistical evaluation showed a significant effect of studfarm-specific environment on IgE levels against the different allergens, but genetic factors also influenced allergen-specific IgE production: an heritability of 0.33 was found for IgE levels ...
Hodgkinson JE, Freeman KL, Lichtenfels JR, Palfreman S, Love S, Matthews JB.The efficacy of five daily fenbendazole (FBZ) treatments was tested against benzimidazole-resistant cyathostomins in naturally infected horses (n=13). Horses were treated with pyrantel embonate (PYR) to remove adult strongyles followed, 7 days later, by a 5-day course of FBZ. The PYR treatment produced an average faecal egg count reduction of 98%. All samples were negative by faecal egg count 7 days after the start of the FBZ treatment. Positive egg counts were observed from 28 days after the start of FBZ treatment and all horses displayed positive faecal egg counts by 77 days after treatment....
Wagner B, Robeson J, McCracken M, Wattrang E, Antczak DF.Recombinant cytokines are valuable tools for functional studies and candidates for vaccine additives or therapeutic use in various diseases. They can also be used to generate specific antibodies to analyze the roles of different cytokines during immune responses. We generated a mammalian expression system for recombinant cytokines using the equine IgG1 heavy chain constant region as a tag for detection and purification of the expressed cytokine, demonstrated here using equine interferon-gamma (IFN-gamma), interleukin-2 (IL-2), interleukin-4 (IL4) and transforming growth factor-beta1 (TGF-beta1...
Hussain KA, Issel CJ, Schnorr KL, Rwambo PM, West M, Montelaro RC.Monoclonal antibodies (MCAbs) were used to dissect the antigenic sites of the surface glycoproteins of the prototype cell-adapted Wyoming strain of equine infectious anemia virus (EIAV). Serologic reactivities of these MCAbs were determined by ELISA, additive ELISA, competitive ELISA, and Western blot assays. The results indicated that antigenic reactivity of gp90 was localized on at least four distinct epitopes, two of which were important in neutralization. Our studies also revealed that these epitopes were localized on overlapping antigenic sites on gp90. On the other hand, only two distinc...
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...
Goubran Botros H, Gregoire C, Rabillon J, David B, Dandeu JP.Horse serum albumin is present in the near vicinity of the animal, while dog and cat serum albumins are very common allergens present in house dust. Human patients clinically defined as allergic to horse could react with horse serum albumin by means of IgE or IgG antibodies. Studies regarding the specificities of these antibodies by inhibition enzyme-linked immunosorbent assay (ELISA) and depletion experiments have demonstrated that they are directed against dog serum albumin and cross-react not only with horse serum albumin but with other serum albumins from different origins. To investigate ...
Ball JM, Henry NL, Montelaro RC, Newman MJ.A simple, versatile and very inexpensive procedure for cross-linking synthetic peptides to the polystyrene surfaces of micro-well assay plates for use in ELISA was developed. The method is based on the use of poly-L-lysine (PLL) as the anchor protein for synthetic peptides which were then easily and covalently linked to the PLL using glutaraldehyde. The synthetic peptides used for the study were based on the amino acid sequence of the equine infectious anemia virus (EIAV) envelope sequence and evaluated as antigens in an ELISA designed to detect antibodies in serum of EIAV-infected horses and ...
