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.
Galvin P, Gildea S, Arkins S, Walsh C, Cullinane A.Antibodies against equine influenza virus (EIV) are traditionally quantified by haemagglutination inhibition (HI) or single radial haemolysis (SRH). Objective: To evaluate an ELISA for the detection of antibodies against influenza nucleoprotein in the diagnosis and surveillance of equine influenza (EI). Methods: The ELISA was compared with the SRH and HI tests. Serial serum samples from 203 naturally and 14 experimentally infected horses, from 60 weanlings following primary vaccination with five different vaccines (two whole inactivated vaccines, two ISCOM-based subunit vaccines and a recombin...
Ginel PJ, Hernández E, Lucena R, Blanco B, Novales M, Mozos E.Insect bite hypersensitivity (IBH) is a common cause of pruritus in horses, but there are few controlled studies on the efficacy of allergen-specific immunotherapy (ASIT). Atopic dermatitis and IBH can present with overlapping clinical signs; multiple insect and environmental allergens could be indicated in these horses to achieve effective hyposensitization. Although the success of ASIT using Culicoides spp. whole-body extracts is controversial, there are no controlled studies published that clearly show benefit from this form of therapy. Objective: The objective was to evaluate the efficacy ...
Chung C, Wilson C, Timoney P, Balasuriya U, Adams E, Adams DS, Evermann JF, Clavijo A, Shuck K, Rodgers S, Lee SS, McGuire TC.The objective of the present study was to validate a previously described competitive enzyme-linked immunosorbent assay (cELISA) to detect antibody to Equine arteritis virus (EAV) based on GP5-specific nonneutralizing monoclonal antibody (mAb) 17B7(9) using the World Organization for Animal Health (OIE)-recommended protocol, which includes the following 5 in-house analyses. 1) The assay was calibrated with the OIE-designated reference serum panel for EAV; 2) repeatability was evaluated within and between assay runs; 3) analytical specificity was evaluated using sera specific to related viruses...
Ranjithkumar M, Saravanan BC, Yadav SC, Kumar R, Singh R, Dey S.Trypanosoma evansi infection typically produces wasting disease, but it can also develop into a neurological or meningoencephalitis form in equids. Trypanosomiasis in horses was treated with quinapyramine sulfate, and all the 14 infected animals were recovered clinically. After clinical recovery, four animals developed a neurological form of the disease at various intervals. Two of these animals treated with diminazene aceturate recovered temporarily. Repeated attempts failed to find the parasite in the blood or the cerebrospinal fluid (CSF), but all of the animals were positive in enzyme-link...
Pearl CA, Roser JF.Lactoferrin is one of the most abundant proteins secreted by the stallion epididymis, but its cellular localization and regulation remain unknown. This study was designed to address the following objectives: (1) identify the epididymal cell types producing lactoferrin in pre-pubertal, peri-pubertal and post-pubertal animals; (2) demonstrate that lactoferrin binds to stallion sperm; and (3) determine if testosterone and estradiol regulate lactoferrin secretion in vitro. Using an immunohistochemical method, lactoferrin was localized in the cytoplasm of principal cells in the corpus and cauda of ...
Vieira TS, Vieira RF, Finger MA, Nascimento DA, Sicupira PM, Dutra LH, Deconto I, Barros-Filho IR, Dornbusch PT, Biondo AW, Vidotto O.The objective of this study was to evaluate the seroepidemiological data of Babesia caballi and Theileria equi in horses from a rural settlement and carthorses from urban areas of Paraná State, southern Brazil. A total of 198 horses, including 32 from the rural settlement and 166 carthorses from Colombo (n=48), Pinhais (n=76), Londrina (n=24), and Curitiba city (n=18) was sampled and tested using a commercial competitive inhibition ELISA (cELISA) test. Out of the 198 horses, 193 (97.5%) were seropositive for at least one piroplasm species. Antibodies to T. equi were detected in 155/198 horses...
Pittaway CE, Lawson AL, Coles GC, Wilson AD.Infection of horses with Anoplocephala perfoliata induces a severe inflammatory reaction of the caecal mucosa around the site of parasite attachment adjacent to the ileocecal valve. Lesions show epithelial erosion or ulceration of the mucosa with infiltration by eosinophils, lymphocytes and mast cells leading to oedema, gross thickening and fibrosis of the caecal wall. Despite this evidence of an inflammatory reaction to A. perfoliata within the mucosa of the caecum there is little information about the nature of the local immune response to A. perfoliata. An ELISA which assays serum IgG(T) an...
Pivoto FL, de Macêdo AG, da Silva MV, Ferreira FB, Silva DA, Pompermayer E, Sangioni LA, Mineo TW, Vogel FS.Protozoa from the family Sarcocystidae are agents of reproductive and neurological disorders in horses. The transmission of these protozoa may occur via horizontal or vertical means, and the frequency and potential of the later is not fully elucidated in horses. Thus, the aim of study was to correlation levels of antibodies in mares with pre colostral foals seropositive and assess the level and distribution of antibodies against Neospora spp., Sarcocystis neurona and Toxoplasma gondii, in mares and pre colostral foals at the parturition. The blood samples were collected from mares immediately ...
Mazzei M, Savini G, Di Gennaro A, Macchioni F, Prati MC, Guzmàn LR, Tolari F.West Nile virus (WNV) is a mosquito-borne virus belonging to the family Flaviviridae included in the Japanese encephalitis antigenic complex (JEAC). A seroepidemiological study was carried out in 2011 using 160 horse sera collected from different areas of Bolivia to investigate the presence of WNV antibody. A high proportion (59.4%) of the tested sera were positive to a commercially available WNV competitive enzyme-linked immunosorbent assay (C-ELISA). Sixty-six randomly selected C-ELISA-positive sera were further tested by WNV plaque reduction neutralization test (PRNT), virus neutralization ...
Merl J, Deeg CA, Swadzba ME, Ueffing M, Hauck SM.Most autoimmune diseases are multifactorial diseases and are caused by the immunological reaction against a number of autoantigens. Key for understanding autoimmune pathologies is the knowledge of the targeted autoantigens, both initially and during disease progression. We present an approach for autoantigen identification based on isolation of intact autoantibody-antigen complexes from body fluids. After organic precipitation of high molecular weight proteins and free immunoglobulins, released autoantigens were identified by quantitative label-free liquid chromatography mass spectrometry. We ...
Szóstek AZ, Lukasik K, Galvão AM, Ferreira-Dias GM, Skarzynski DJ.The aim of the study was to characterize endometrial mRNA transcription, immunolocalization, and protein expression of interleukin (IL) 1alpha, IL1beta, IL6, and IL1RI, IL1RII, and IL6Ralpha/beta in the course of endometrosis during the estrous cycle. Additionally, the influence of IL1alpha, IL1beta, and IL6 on prostaglandin (PG) secretion and PG synthase mRNA transcription in endometrial tissue during endometrosis was investigated. The endometrial samples were obtained at the early (n = 12), mid- (n = 12), and late (n = 12) luteal phases and at the follicular (n = 12) phase of the estrous cyc...
Ziegler U, Skrypnyk A, Keller M, Staubach C, Bezymennyi M, Damiani AM, Osterrieder N, Groschup MH.West Nile virus (WNV) is a mosquito-borne virus of global importance. Over the last two decades, it has been responsible for significant numbers of cases of illness in humans and animals in many parts of the world. In Ukraine, WNV infections in humans and birds were first reported more than 25 years ago, yet the current epidemiological status is quite unclear. In this study, serum samples from over 300 equines were collected and screened in order to detect current WNV activity in Ukraine with the goal to estimate the risk of infection for humans and horses. Sera were tested by enzyme-linked im...
Toishi Y, Tsunoda N, Tagami M, Hashimoto H, Kato F, Suzuki T, Nagaoka K, Watanabe G, Tokuyama S, Okuda K, Taya K.Evaluation of a new chemiluminescent enzyme immunoassay, the PATHFAST assay system (PATHFAST), for measurement of circulating progesterone in mares was performed. Five mares at the mid-luteal stage were administrated a single i.m. injection of prostaglandin F2α analog (PGF2α; cloprostenol 250 μg/ml), and then blood samples were collected from the jugular vein at 0, 15, 30 and 45 min, at one-hour intervals until 24 and at 48 hr via a catheter in the jugular vein. To monitor the physiological changes in circulating progesterone in mares after induced luteolysis, concentrations of progesterone...
Grosche A, Morton AJ, Graham AS, Polyak MM, Freeman DE.To determine the effect of large colon ischemia and reperfusion on concentrations of the inflammatory neutrophilic protein calprotectin and other clinicopathologic variables in jugular and colonic venous blood in horses. Methods: 6 healthy horses. Methods: Horses were anesthetized, and ischemia was induced for 1 hour followed by 4 hours of reperfusion in a segment of the pelvic flexure of the large colon. Blood samples were obtained before anesthesia, before induction of ischemia, 1 hour after the start of ischemia, and 1, 2, and 4 hours after the start of reperfusion from jugular veins and ve...
Yeargan MR, Alvarado-Esquivel C, Dubey JP, Howe DK.Equine protozoal myeloencephalitis (EPM) is a debilitating disease of horses caused by Sarcocystis neurona and Neospora hughesi. Sera from 495 horses in Durango State, Mexico were tested for anti-protozoal antibodies using enzyme-linked immunosorbent assays (ELISAs) based on major surface antigens of these two parasites. Antibodies to S. neurona were detected in 240 (48.5%) of the 495 horse sera tested with the rSnSAG2/4/3 trivalent ELISA. Multivariate analysis showed that exposure to S. neurona was associated with age, feeding grains and crops, and small herd size. Antibodies to N. hughesi we...
Kumar S, Kumar R, Gupta AK, Yadav SC, Goyal SK, Khurana SK, Singh RK.Equine piroplasmosis is a tick-transmitted protozoan disease caused by Theileria equi and/or Babesia caballi. In the present study, we expressed a 53kDa protein from the truncated EMA-2 gene of T. equi (Indian strain) and developed EMA-2ELISA using this expressed protein. This ELISA is able to detect T. equi-specific antibodies in experimentally infected animals as early as 9 days post-infection. The assay developed was validated with the OIE recommended competitive ELISA (cELISA) on 120 serum samples and significant agreement (kappa=0.93) was observed between results of both the ELISAs which ...
Ometto T, Durigon EL, de Araujo J, Aprelon R, de Aguiar DM, Cavalcante GT, Melo RM, Levi JE, de Azevedo Júnior SM, Petry MV, Neto IS, Serafini P....West Nile virus (WNV) is an emergent pathogen that is widely distributed in North and Central America. The recent introduction in South America has focused attention on the spread of WNV across Southern American countries. The transmission network involves mosquitoes, birds, horses and humans. Methods: The serological evaluation of sera from 678 equids and 478 birds was performed using a WNV-specific blocking ELISA, and only the positive results were confirmed by plaque reduction neutralisation tests (PRNTs). Molecular analysis was performed on sera from 992 healthy equids and on 63 macerates ...
Reed SM, Howe DK, Morrow JK, Graves A, Yeargan MR, Johnson AL, MacKay RJ, Furr M, Saville WJ, Williams NM.Recent work demonstrated the value of antigen-specific antibody indices (AI and C-value) to detect intrathecal antibody production against Sarcocystis neurona for antemortem diagnosis of equine protozoal myeloencephalitis (EPM). Objective: The study was conducted to assess whether the antigen-specific antibody indices can be reduced to a simple serum : cerebrospinal fluid (CSF) titer ratio to achieve accurate EPM diagnosis. Methods: Paired serum and CSF samples from 128 horses diagnosed by postmortem examination. The sample set included 44 EPM cases, 35 cervical-vertebral malformation (CVM) ca...
Horsington J, Hartley CA, Gilkerson JR.Respiratory infections are a major burden in the performance horse industry. Equine rhinitis B virus (ERBV) has been isolated from horses displaying clinical respiratory disease, and ERBV-neutralizing antibodies have been detected in 50-80% of horses in reported surveys. Current ERBV isolation and detection methods may underestimate the number of ERBV-positive animals and do not identify multiple serotype infections. The aim of the current study was to develop a serotyping ERBV antibody-detection enzyme-linked immunosorbent assay (ELISA) and examine the seroprevalence of ERBV in a group of Aus...
Singha H, Goyal SK, Malik P, Khurana SK, Singh RK.Equine infectious anemia (EIA)-a retroviral disease caused by equine infectious anemia virus (EIAV)-is a chronic, debilitating disease of horses, mules, and donkeys. EIAV infection has been reported worldwide and is recognized as pathogen of significant economic importance to the horse industry. This disease falls under regulatory control program in many countries including India. Control of EIA is based on identification of inapparent carriers by detection of antibodies to EIAV in serologic tests and "Stamping Out" policy. The current internationally accepted test for diagnosis of EIA is the ...
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...
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...
Ziegler U, Angenvoort J, Klaus C, Nagel-Kohl U, Sauerwald C, Thalheim S, Horner S, Braun B, Kenklies S, Tyczka J, Keller M, Groschup MH.West Nile virus (WNV) is a mosquito-borne viral pathogen of global importance and is considered to be the most widespread flavivirus in the World. Horses, as dead-end hosts, can be infected by bridge mosquito vectors and undergo either subclinical infections or develop severe neurological diseases. The aim of this study was to detect WNV specific antibodies in horses in Germany as an indicator for an endemic circulation of WNV. Sera from more than 5,000 horses (primarily fallen stock animals) were collected in eight different federal states of Germany from 2010 to 2012. Sera were screened by a...
Gilliam LL, Ownby CL, McFarlane D, Canida A, Holbrook TC, Payton ME, Krehbiel CR.Rattlesnake bites in horses are not uncommon and the clinical outcomes are widely variable. Treatment of horses with anti-venom is often cost prohibitive and could have negative consequences; therefore, the development of a quantitative test to determine if anti-venom therapy is indicated would be valuable. The objective of this study was to develop an ELISA to detect rattlesnake venom in biological samples from clinically bitten horses. Nineteen horses were enrolled in the study. Urine was available from 19 horses and bite site samples were available from 9 horses. A double sandwich fluoresce...
Barbic L, Vilibic-Cavlek T, Listes E, Stevanovic V, Gjenero-Margan I, Ljubin-Sternak S, Pem-Novosel I, Listes I, Mlinaric-Galinovic G, Di Gennaro A....We report the first serological evidence of Usutu virus (USUV) infection in horses in Croatia. During 2011, 1380 horse serum samples from healthy animals were collected from six northern Croatian counties. All samples were first screened for West Nile virus (WNV) immunoglobulin G (IgG) antibodies using an enzyme-linked immunosorbent assay (ELISA). Sixty-nine WNV ELISA-reactive samples were further tested for WNV antibodies by a virus neutralization assay (VN assay) and plaque reduction neutralization test (PRNT), and USUV by a VN assay and tick-borne encephalitis virus (TBEV) antibodies by PRN...
Yan W, Saleem MH, McDonough P, McDonough SP, Divers TJ, Chang YF.Leptospira immunoglobulin (Ig)-like (Lig) proteins are a novel family of surface-associated proteins in which the N-terminal 630 amino acids are conserved. In this study, we truncated the LigA conserved region into 7 fragments comprising the 1st to 3rd (LigACon1-3), 4th to 7.5th (LigACon4-7.5), 4th (LigACon4), 4.5th to 5.5th (LigACon4.5-5.5), 5.5th to 6.5th (LigACon5.5-6.5), 4th to 5th (LigACon4-5), and 6th to 7.5th (LigACon6-7.5) repeat domains. All 7 recombinant Lig proteins were screened using a slot-shaped dot blot assay for the diagnosis of equine leptospirosis. Our results showed that Li...
Chinikar S, Shah-Hosseini N, Mostafavi E, Moradi M, Khakifirouz S, Jalali T, Goya MM, Shirzadi MR, Zainali M, Fooks AR.This study was undertaken to determine the seroprevalence of West Nile virus (WNV) in human and equine sera in Iran. Blood samples were tested from 300 human samples and 315 equine samples in five geographic zones of north and central parts of Iran between 2010 and 2012. All samples were tested for the immunoglobulin G (IgG) antibody to WNV by using an enzyme-linked immunosorbent assay (ELISA). Of all samples, 4 (1.3%) human and 9 (2.8%) equines were considered to be seropositive for WNV. These results suggest circulation and exposure of the human and equine populations to WNV in Iran.
Diab SS, Rodriguez-Bertos A, Uzal FA.Clostridium difficile is commonly associated with diarrhea and colitis in humans and other mammals, including horses. To this date, the epidemiologic, microbiologic, clinical, and diagnostic aspects of C. difficile-associated disease (CDAD) in horses have been thoroughly described. However, reports describing the enteric pathology of this disease in horses are limited. This study presents a comprehensive description of the pathologic characteristics of CDAD in 21 horses and discusses the criteria for the diagnosis of the disease. Case selection was based on C. difficile A/B toxins detection (e...
Wagnerová P, Sak B, Květoňová D, Maršálek M, Langrová I, Kváč M.A total of 9 (8 stallions and 1 mare) 1 year old ponies were used for the experimental infection caused by Encephalitozoon cuniculi genotype II (10(7) spores per animal). Subsequently, individual horses were slaughtered 7, 14, 21, 28, 35, 42, 49, 56, and 63 days post infection. Immediately after slaughter, tissues samples of stomach, duodenum, jejunum, ileum, caecum, colon, spleen, liver, kidney, bladder, heart, lungs, and brain were sampled. In addition, urine, feces and blood specimens were collected. Enzyme-linked immunosorbent assay was used for determination of humoral immune response and...
Makra Z, Tuboly T, Bodó G.The aim of this study was to describe long-term follow-up and difference in immune reactions in the tear film following penetrating keratoplasty (PK) in horses when differently preserved corneas were utilised. This report describes for the first time the use of corneal grafts preserved in tissue culture media in equine PK. Eight experimental horses with normal eyes were included and freshly harvested, frozen or preserved corneal grafts were used for the PK. The graft-taking technique and storage, PK surgery, postoperative treatments and complications are described. The mean postoperative follo...
Matsushita T, Hesterberg LK, Porter JP, Smith BJ, Newman LE.Two diagnostic tests are approved for detecting antibody to equine infectious anemia virus: the agar-gel immunodiffusion (AGID) test and the competitive enzyme-linked immunosorbent assay (ELISA). A total of 420 sera from National Veterinary Services Laboratories check sets were tested with the AGID and competitive ELISA. A 100% correlation was obtained. The AGID and competitive ELISA were further used to test difficult samples with low levels of equine infectious anemia antibody (weak positives). A third test (Western blot) was also used with these weak positive samples to resolve any discorda...
Berxholi K, Ziegler U, Rexhepi A, Schmidt K, Mertens M, Korro K, Cuko A, Angenvoort J, Groschup MH.Serum samples collected from 167 equines of 12 districts in Albania were tested for West Nile virus-specific antibodies by enzyme-linked immunosorbent assay and virus neutralization assay, using WNV lineage 1 and 2. In addition, 95 bird serum samples from Albania and 29 horse samples from Kosovo were tested in ELISA. An overall seroprevalence rate of 22% was found in horses from Albania, whereas no specific antibodies were found in the equine samples from Kosovo and the bird samples. This is the first report indicating WNV infections in animals in Albania, and the first reported seroprevalence...
Cook RF, Sinclair R, Mumford JA.An antigen capture indirect enzyme linked immunosorbent assay (ELISA) was developed to detect influenza nucleoprotein antigen in nasal secretions from horses infected with A/equine/H3N8 viruses. Results from this assay were compared with conventional virus isolation in embryonated hens eggs.
Wang M, Guo W, Igarashi I, Xuan X, Wang X, Xiang W, Jia H.The objective of this study is to investigate the seroprevalence of equine piroplasmosis in China. A total of 1990 sera were collected from clinically healthy horses in various districts located in ten different provinces of China and examined by using indirect enzyme-linked immunosorbent assays (ELISAs) with recombinant Theileria equi (T. equi) merozoite antigen 2 (rEMA-2) and Babesia caballi (B. caballi) 48-kDa rhoptry protein (rBc48), respectively. The results showed that 1,018 (51.16%) and 229 (11.51%) samples were positive for B. caballi and T. equi infection, respectively. The number of ...
Gordon SJG, Bolwell C, Rogers CW, Musuka G, Kelly P, Guthrie A, Mellor PS, Hamblin C.Sentinel herds and samples submitted by private equine practitioners were used to determine the sero-prevalence and sero-incidence of African horse sickness virus (AHSV) and equine encephalosis virus (EEV) in horse and donkey populations in the Highveld region of Zimbabwe. The sero-prevalence and sero-incidence of antibodies against these viruses were determined using the competitive enzyme-linked immunosorbent assay (ELISA) for the detection of serum antibodies. In donkeys, the median sero-prevalence of AHSV antibodies, across the three rainy seasons under study, was 75% (inter quartile...
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...
Dutta SK, Mattingly BL, Shankarappa B.The antibody response and the antibody reactivity to component antigens of Ehrlichia risticii were studied in horses with induced Potomac horse fever. These horses had no detectable antibodies to E. risticii in their preinoculation (PrI) sera by indirect fluorescent-antibody assay and enzyme-linked immunosorbent assay (ELISA). All the horses exhibited typical disease features following experimental infection and responded with specific antibodies, as measured by ELISA and indirect fluorescent-antibody assay. A primary antibody response was detected in 70% of the horses, while a secondary-type ...
Nielsen MK, Scare J, Gravatte HS, Bellaw JL, Prado JC, Reinemeyer CR.Strongylus vulgaris is the most pathogenic nematode parasite of horses. Its extensive migration in the mesenteric blood vessels can lead to life-threatening intestinal infarctions. Recent work has shown that this parasite is still identified among managed horse populations. A serum enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of migrating larvae of S. vulgaris. Previous work has documented an increase in ELISA values following larvicidal treatment with ivermectin and suggested that the target parasite antigen is primarily produced by the later larval stages. T...
Kirkland PD, Delbridge G.An important consideration in the selection of a vaccine during the Australian equine influenza (EI) outbreak in 2007 was the ability to differentiate between infected and vaccinated animals (DIVA). A blocking enzyme-linked immunosorbent assay (bELISA) targeted for the nucleoprotein of influenza A viruses was developed to differentiate between naturally infected horses and horses vaccinated with the ProteqFlu® vaccine, which only induces a response to viral haemagglutinin. This bELISA assay met the DIVA requirements and was used extensively during the EI control and eradication programs and '...
Carvalho GM, Ramos CP, Lobato FCF, Guedes RMC, Giaretta PR, Silva ROS.Despite the known importance of Clostridioides (Clostridium) difficile infection (CDI) in animals, there are no published guidelines for the diagnosis of CDI. The performance of the available commercial methods, all standardized for human stool samples, can vary according to the animal species. Thus, the aim of the present study was to review the literature on the detection of C. difficile in pigs, horses, and dogs. The detection of toxins A and B using enzyme immunoassays seems to have low performance in piglet and dog samples, while it shows high sensitivity for the diagnosis of CDI in foal...
Fall AG, Diaïté A, Etter E, Bouyer J, Ndiaye TD, Konaté L.Active catches of adult females of Aedes vexans arabiensis Patton, (Diptera: Culicidae) Patton by nets or aspirator, were conducted in 2003 and 2004 in the vegetation at the edge of temporary ponds in Barkedji, Senegalese Ferlo area. Two hundred and forty-one engorged females were captured, dissected and the gut content adsorbed on a Whatman filter paper and analysed using the enzyme-linked immunosorbent assay (ELISA) technique to determinate the bloodmeal origin. Results indicated that Ae v. arabiensis fed primarily on mammals, including horses (35.7% of the bloodmeals), but also on birds (10...
Cruz F, Fores P, Ireland J, Moreno MA, Newton R.No cases of equine infectious anaemia (EIA) have been reported in Spain since 1983. Factors that could increase the risk of reintroducing equine infectious anaemia virus (EIAV) into Spain include the recent occurrence of the disease in Europe and the absence of compulsory serological testing before importation into Spain. Objective: Given the importance of the Spanish Purebred (SP) horse breeding industry in Spain, the aim of this cross-sectional study was to provide evidence of freedom from EIAV in SP stud farms in Central Spain. Methods: Serum samples from 555 SP horses, collected between Se...
Verma RD, Sharma JK, Venkateswaran KS, Batra HV.A dot enzyme-linked immunosorbent assay (dot ELISA) was developed for diagnosis of glanders in equines. The test was based on the detection of IgG antibodies to Pseudomonas mallei antigens bound to nitrocellulose coated on plastic strips (dipsticks), the reaction being amplified by an avidin-biotin system with biotinylated anti-horse IgG and horseradish peroxidase-avidin D. Sera from 810 normal, six naturally infected and 48 sensitized equines were tested by this assay, and results were compared with complement fixation, indirect haemagglutination and counter-immunoelectrophoresis tests. Dot E...
House C, Mikiciuk PE, Berninger ML.Five serological methods of diagnosing African horse sickness were evaluated, using a battery of serum samples from experimental horses vaccinated and challenged with each serotype of African horse sickness virus (AHSV1 through AHSV9): agar gel immunodiffusion (AGID), indirect fluorescent antibody (IFA), complement fixation (CF), virus neutralization (VN), and enzyme-linked immunosorbent assay (ELISA). The 5 tests were also compared using a panel of field samples, convalescent equine sera with antibodies to domestic equine viral diseases, and sera from horses awaiting export. The ELISA describ...
Ziegler U, Skrypnyk A, Keller M, Staubach C, Bezymennyi M, Damiani AM, Osterrieder N, Groschup MH.West Nile virus (WNV) is a mosquito-borne virus of global importance. Over the last two decades, it has been responsible for significant numbers of cases of illness in humans and animals in many parts of the world. In Ukraine, WNV infections in humans and birds were first reported more than 25 years ago, yet the current epidemiological status is quite unclear. In this study, serum samples from over 300 equines were collected and screened in order to detect current WNV activity in Ukraine with the goal to estimate the risk of infection for humans and horses. Sera were tested by enzyme-linked im...
Prescott JF, Fernandez AS, Nicholson VM, Patterson MC, Yager JA, Viel L, Perkins G.An enzyme-linked immunosorbent assay (ELISA) was developed against Rhodococcus equi using Triton X-114 detergent extracted whole cell material, in which the virulence associated protein (VapA) predominated. Enzymelinked immunosorbent assay titres corresponded to antibody reacting with VapA on Western blots. There was considerable variation in antibody titres of nonimmunised mares and in the time when the colostrally derived antibody of their foals had declined to low or undetectable titres. In general, antibodies in foals declined to their lowest levels at age 4-8 weeks. Seroconversion occurre...
Parma AE, Fernández AS, Santisteban CG, Bowden RA, Cerone SI.An antigenic relationship between Leptospira interrogans and equine cornea was previously described by us. An enzyme-linked immunosorbent assay was employed in the present work to investigate the existence of anti-leptospira and anti-cornea antibodies in tears, aqueous humor and serum from horses inoculated i.m. with those antigens. Ten days after a booster by the same route, antibodies that bind to microtiter plates, coated with an homogenate of either equine cornea or leptospira, were detected in those fluids and in the sera. At the same time, the corneas of the horses began to develop a dif...
Pearl CA, Roser JF.Lactoferrin is one of the most abundant proteins secreted by the stallion epididymis, but its cellular localization and regulation remain unknown. This study was designed to address the following objectives: (1) identify the epididymal cell types producing lactoferrin in pre-pubertal, peri-pubertal and post-pubertal animals; (2) demonstrate that lactoferrin binds to stallion sperm; and (3) determine if testosterone and estradiol regulate lactoferrin secretion in vitro. Using an immunohistochemical method, lactoferrin was localized in the cytoplasm of principal cells in the corpus and cauda of ...
Nielsen MK, Vidyashankar AN, Gravatte HS, Bellaw J, Lyons ET, Andersen UV.Strongylus vulgaris is regarded as the most pathogenic helminth parasite infecting horses. Migrating larvae cause pronounced endarteritis and thrombosis in the cranial mesenteric artery and adjacent branches, and thromboembolism can lead to ischemia and infarction of large intestinal segments. A recently developed serum ELISA allows detection of S. vulgaris-specific antibodies during the six-month-long prepatent period. A population of horses has been maintained at the University of Kentucky without anthelmintic intervention since 1979, and S. vulgaris has been documented to be highly prevalen...
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...
Kosinska MK, Eichner G, Schmitz G, Liebisch G, Steinmeyer J.The current limitations in evaluating synovial fluid (SF) components in health and disease and between species are due in part to the lack of data on normal SF, because of low availability of SF from healthy articular joints. Our study aimed to quantify species-dependent differences in phospholipid (PL) profiles of normal knee SF obtained from equine and human donors. Knee SF was obtained during autopsy by arthrocentesis from 15 and 13 joint-healthy human and equine donors, respectively. PL species extracted from SF were quantitated by mass spectrometry whereas ELISA determined apolipoprotein ...
Grosche A, Morton AJ, Graham AS, Polyak MM, Freeman DE.To determine the effect of large colon ischemia and reperfusion on concentrations of the inflammatory neutrophilic protein calprotectin and other clinicopathologic variables in jugular and colonic venous blood in horses. Methods: 6 healthy horses. Methods: Horses were anesthetized, and ischemia was induced for 1 hour followed by 4 hours of reperfusion in a segment of the pelvic flexure of the large colon. Blood samples were obtained before anesthesia, before induction of ischemia, 1 hour after the start of ischemia, and 1, 2, and 4 hours after the start of reperfusion from jugular veins and ve...
Smith MR, Wright IM, Minshall GJ, Dudhia J, Verheyen K, Heinegård D, Smith RK.To evaluate digital flexor tendon sheath (DFTS) synovial fluid cartilage oligomeric matrix protein (COMP) concentrations as a molecular marker for intrathecal pathology. Methods: Case control study. Methods: Horses (n=46) with DFTS tenosynovitis; 23 fresh cadaver horses. Methods: DFTS synovial fluid samples were collected from clinical cases with noninfected DFTS tenosynovitis and from control DFTS. Clinical and surgical findings were recorded, and dissection of control limbs was performed to confirm the DFTS to be grossly normal. Synovial fluid COMP was quantified using a homologous competiti...
Warnken T, Delarocque J, Schumacher S, Huber K, Feige K.Insulin dysregulation (ID) with basal or postprandial hyperinsulinemia is one of the key findings in horses and ponies suffering from the equine metabolic syndrome (EMS). Assessment of ID can easily be performed in clinical settings by the use of oral glucose challenge tests. Oral glucose test (OGT) performed with 1 g/kg bodyweight (BW) glucose administered via naso-gastric tube allows the exact administration of a defined glucose dosage in a short time. However, reliable cut-off values have not been available so far. Therefore, the aim of the study was to describe variations in insulin respo...
Alkharsah KR, Al-Afaleq AI.This study was designed to investigate the seroprevalence of WNV antibodies in humans, horses, and pigeons in the Eastern Province of Saudi Arabia. Methods: Blood samples were collected from 323 humans, 147 horses, and 282 pigeons from two regions, Al-Ahsa and Al-Qatif, in East of Saudi Arabia. Serum samples were tested for anti-WNV antibodies by ELISA. Results: The percentage of anti-WNV antibodies in the human population was found to be 9.6% (3.1% in females and 6.5% in males). This percentage was much higher in horses, as 71.4% (105/147) of the horses had anti-WNV antibodies. However, no st...
Hartley CA, Wilks CR, Studdert MJ, Gilkerson JR.To compare methods of detecting equine herpesvirus type 1 (EHV1)- and EHV4-specific antibodies in horse sera. Methods: 33 acute and convalescent serum samples from experimentally or naturally infected horses after confirmed EHV1 or EHV4 infection. Methods: For each sample, serum antibody titers against EHV1 and EHV4 were determined by use of virus neutralization (VN) and complement fixation (CF) assays. The ELISA absorbance values for each serum sample were determined against the EHV1 and EHV4 recombinant ELISA antigens. Values obtained for acute and convalescent sera in each assay were compar...
Aguilar-Montes de Oca S, Montes-de-Oca-Jiménez R, Carlos Vázquez-Chagoyán J, Barbabosa-Pliego A, Eliana Rivadeneira-Barreiro P....Peptides constitute an alternative and interesting option to develop treatments, vaccines, and diagnostic tools as they demonstrate their scope in several health aspects; as proof of this, commercial peptides for humans and animals are available on the market and used daily. This review aimed to know the role of peptides in the field of veterinary diagnosis, and include peptide-based enzyme-linked immunosorbent assay (pELISA), lateral flow devices, and peptide latex agglutination tests that have been developed to detect several pathogens including viruses and bacteria of health and production ...
Huang X, Xuan X, Verdida RA, Zhang S, Yokoyama N, Xu L, Igarashi I.An immunochromatographic test for the simultaneous detection of Babesia caballi- and B. equi-specific antibodies (BceICT) was developed using a recombinant B. caballi 48-kDa rhoptry protein (rBc48) and a recombinant truncated B. equi merozoite antigen 2 (rEMA-2t). An evaluation of the ability of the BceICT to detect antibodies in sera from uninfected horses and experimentally infected horses showed high sensitivities and specificities of 83.3% (10/12 sera) and 92.9% (52/56 sera), respectively, for the anti-B. caballi antibody and 94.1% (16/17 sera) and 88.2% (45/51 sera), respectively, for the...
Rodríguez M, Joseph S, Pfeffer M, Raghavan R, Wernery U.African horse sickness (AHS) is a serious viral disease of equids resulting in the deaths of many equids in sub-Saharan Africa that has been recognized for centuries. This has significant economic impact on the horse industry, despite the good husbandry practices. Currently, prevention and control of the disease is based on administration of live attenuated vaccines and control of the arthropod vectors. Results: A total of 29 horses in 2 groups, were vaccinated. Eighteen horses in Group 1 were further divided into 9 subgroups of 2 horses each, were individually immunised with one of 1 to 9 AHS...
Divers TJ, Grice AL, Mohammed HO, Glaser AL, Wagner B.Changes in ELISA serology are frequently used to determine antibiotic treatment success for Lyme disease in horses. This concept was based upon a previous report showing a marked decline in ELISA values in experimentally infected and antibiotic-treated ponies. Changes in Lyme serology following antibiotic treatment in naturally infected horses have not been reported. The objective of this study was to compare Borrelia ELISA antibody concentrations in naturally exposed horses both before and following antibiotic treatment for Lyme disease. A retrospective study was performed comparing oxytetrac...
