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.
Takai S, Morishita T, Nishio Y, Sasaki Y, Tsubaki S, Higuchi T, Hagiwara S, Senba H, Kato M, Seno N.We recently generated a monoclonal antibody immunoglobulin G1 (MAb 10G5), which can recognize 15- to 17-kDa antigens, virulence-associated antigens of Rhodococcus equi, and developed a colony blot enzyme-linked immunosorbent assay with MAb 10G5 for the rapid identification of virulent R. equi. In this epidemiologic study, we evaluated the results of the colony blot test in the identification of virulent isolates of R. equi from feces of horses and soil and compared them with those from a conventional procedure (plasmid profiles of isolates by agarose gel electrophoresis). Environmental isolate...
Scudamore CL, Pemberton AD, Miller HR, McDonnell AM, Thomson SR, Dawson A, Watson ED.An enzyme linked immunosorbent assay (ELISA) was developed and used to estimate the concentrations of the serine proteinase inhibitor, alpha-1 proteinase inhibitor (API), in uterine flushings recovered from mares at different stages of the oestrous cycle and before and after the induction of experimental endometritis. There was a significant increase in the concentrations of API and albumin relative to total protein in flushings recovered during oestrus compared with dioestrus but no difference was observed in the concentrations of these proteins relative to total protein before and after the ...
Jaspers U, Thiele D, Krauss H.A competitive ELISA system for the detection of antibodies against Coxiella (C.) burnetii in cattle, sheep, goats, horses and humans is described. The ELISA is based on a biotinylated monoclonal antibody with specificity for C. burnetii lipopolysaccharide in combination with streptavidin peroxidase. For evaluation and statistical analysis, 413 sera from cattle, sheep, goats, horses and humans were tested in parallel in the indirect immunofluorescence test (IFT). Furthermore, a total of 448 bovine and human sera were also tested with an indirect ELISA and 47 sheep sera were investigated using t...
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 ...
Kihurani DO, Nantulya VM, Mbiuki SM, Mogoa E, Nguhiu-Mwangi J, Mbithi PM.Equines are particularly susceptible to infection with Trypanosoma evansi and T. brucei, but rarely is natural T. congolense and T. vivax infection seen in horses. An outbreak of trypanosomosis occurred in a herd of horses used for patrolling the pineapple fields on the Del Monte Farm, Thika, Kenya initially involving 6 horses. On subsequent screening of the entire group, T. brucei, T. congolense and T. vivax infections were detected in 16 of the 35 horses. The tests used for diagnosis included microscopic examination of stained blood smears, buffy coat technique, mouse inoculation and antigen...
Böse R, Peymann B.From Babesia caballi in vitro cultures a preparation of 100% infected erythrocytes was obtained. From this, B. caballi antigens were extracted with the detergent 3-[(3-Cholamidopropyl)-dimethylammonio]-1-propane-sulfonate (CHAPS) and used as ELISA antigens. A control antigen of normal erythrocytes from the same donor horse was prepared in an identical manner. The ELISA and Western blot were validated by testing of sera from horses experimentally infected with B. caballi or B. equi or not infected with Babesia spp. ELISA and Western blot results were compared with those obtained by the immunofl...
Pompa G, Caloni F, Montana M, Pasqualucci C.1. Isoxsuprine [1-(4-hydroxyphenyl)-2-(1-methyl-2-phenoxyethylamino)-1- propanol] serum concentrations after single- and multiple-dose administration to horse were investigated using immunoenzymatic ELISA, HPLC-UV and thermospray HPLC-MS methods. 2. Using HPLC-MS, isoxsuprine was detected up to 72 h after a single administration (1.2 mg/kg by gastric probe) and up to 96 h after the end of serial administration (1.2 mg/kg every 12 h for 7 days). 3. ELISA detected the drug up to 96 h after a single dose and up to 6 days after the end of prolonged administration. 4. Isoxsuprine is present in hors...
Böse R, Peymann B, Barbosa IP.Sera from 60 horses held in breeding herd in Brazil were examined monthly by ELISA, immunofluorescence antibody test (IFAT) and Western blot. All foals had maternal antibodies detectable by ELISA and IFAT, and sero-conversion took place between the 2nd and 5th month of age. The 48 and 50 kDa antigens were recognized first in the course of infection. Of 79 sera taken after sero-conversion 78 reacted with the 48 kDa antigen, 76 with the 50 kDa, 50 with the 70 kDa, 54 with the 112 kDa, 72 with the 141 kDa antigen. In general, sera from horses older than 1 year reacted with all 5 diagnostic antige...
Stanley S, Wood T, Goodman JP, Henry PA, Woods WE, Chang SL, Tai HH, Watt D, Kwiatkowski S, Blake JW.We have raised antibodies and developed one-step enzyme-linked immunosorbent assays (ELISA) for the diuretics ethacrynic acid and bumetanide as part of a panel of pre- and post-race tests for high potency drugs in racing horses. These ELISA tests are rapid (completed within one hour), sensitive, and can be read by eye. The ELISA detects ethacrynic acid at a drug concentration for half-maximal inhibition (I-50) of about 2.5 ng/mL for the parent drug. After dosing horses intravenously with 5 mg ethacrynic acid per horse, the parent drug or its metabolites are detectable in urine for at least 8 h...
Hagedorn HW, Schulz R, Jaeschke G.An enzyme linked immunosorbent assay (ELISA) was developed to detect the anabolic steroid boldenone in equine blood and urine. The polyclonal antiserum was raised in rabbits, employing boldenone-17-hemisuccinate-bovine serum albumin as antigen. Boldenone-17-hemisuccinate-horseradish peroxidase served as enzyme conjugate. Sensitivity of the assay was 26.0 +/- 3.0 pg/well. Among the endogenous steroids tested only progesterone and testosterone exhibited moderate cross-reactivities, 3.4 and 2.5%, respectively. These cross-reactivities are of no importance for the boldenone assay. For the reductio...
Jakob HP, Eckert J, Jemmi T, Gottstein B.For many decades trichinellosis has not been reported among Swiss domestic pigs. Considering the fact that Trichinella occurs in a sylvatic cycle in Switzerland, a study was designed to reevaluate the present epidemiologic situation by investigating 10,904 fattening pigs, 218 pigs with free access to pasturage or being kept on an alp, 104 domestic boars, 106 horses, 44 wild boars and 538 foxes using a direct and an indirect diagnostic technique (digestion method and serology with ELISA and an excretory/secretory antigen, respectively). The digestion method was performed according to EC-guideli...
Sugiura T, Nakamura H.The zinc content in platelets from rabbits, humans and horses was determined, and the levels of zinc were found to be significantly higher (3 micrograms/10(10) cells) than those in other peripheral blood cells. About 70% of the zinc in the supernatants of platelet lysates could be detected. From the results of gel filtration analysis, the zinc in platelet lysates was found to be bound with a low-molecular-weight protein (MW 6,000-8,000) detected as metallothionein (MT) on the basis of antigenic properties determined by enzyme-linked immunoassay and immunoblotting analysis using monoclonal anti...
Schumacher J, Spano JS, Wilson RC, DeGraves FJ, Duran SH, Ruffin DC.The pharmacokinetic properties of intravenously administered caffeine were studied in 10 horses using a commercially available automated enzyme immunoassay. The harmonic mean for the distribution half-life was 5.2 min (range 1.4-18.7). The harmonic mean for the elimination half-life was 10.18 h (range 6.82-20.92). The harmonic mean of the volume of distribution was 0.32 L/kg (range 0.22-0.53). There was no correlation between the dose of caffeine/kg body weight and the elimination half-life (Spearman's coefficient of rank correlation = 0.19).
Sahu SP, Alstad AD, Pedersen DD, Pearson JE.Immunoglobulin M (IgM) and G (IgG) capture enzyme-linked immunosorbent assays (ELISAs) were used as possible adjuncts to hemagglutination inhibition (HI) and virus neutralization (VN) tests to differentiate between reaction to recent exposure to eastern equine encephalomyelitis (EEE) virus and those due to prior vaccination. Serum samples were evaluated by the IgM-capture ELISA, and the results were compared with those of HI and VN tests. Of 381 serum samples, 51% (195 samples) were positive by HI test (> or = 1:40) and 54% (205 samples) were positive by VN test (> or = 1:10), but only 3...
Livesay GJ, O'Neill T, Hannant D, Yadav MP, Mumford JA.In July 1989 influenza A/equine-2 (H3N8) was isolated from a nasopharyngeal swab taken from a non-thoroughbred horse exhibiting acute clinical respiratory disease. This was the first isolation of equine influenza virus in the United Kingdom since 1981. Subsequent investigations of acute respiratory disease in horses indicated that the infection was dispersed throughout the UK. However, unlike the previous epidemic of 1979, the first horses from which the virus was isolated had been vaccinated. This outbreak of influenza provided an opportunity to evaluate an antigen capture ELISA, directed aga...
Halliwell RE, McGorum BC, Irving P, Dixon PM.An enzyme-linked immunosorbent assay (ELISA) was used to quantify isotype-specific antibody to Micropolyspora faeni and to Aspergillus fumigatus in the sera and bronchoalveolar lavage fluid (BALF) of normal horses, horses with chronic obstructive pulmonary disease (COPD) and horses with other chronic respiratory diseases. Elevated antibody levels were not detected in the sera of affected horses. However, both IgE and IgA antibody to both allergens was significantly elevated in BALF in COPD affected horses sampled both when symptomatic and asymptomatic. Elevated levels were also found in animal...
Orino K, Yamamoto S, Watanabe K.Lower apparent concentrations of ferritin were observed in horse plasma than in serum using the enzyme-linked immunosorbent assay (ELISA). However, the ferritin concentrations in plasma and serum were increased to the same level on heating the samples at 75 degrees C for 15 min. These results suggest that horse plasma has specific ferritin-binding protein(s) which inhibit(s) the ferritin assay. The apparent ferritin concentrations in horse serum were markedly decreased by adding horse fibrinogen to the serum. It was also found that fibrinogen bound to spleen ferritin and inhibited the immunoas...
Nishita T, Goto T, Kimura H, Asari M.1. In this study, carbonic anhydrase III (CA-III) content in 18 equine muscles was determined by enzyme immunoassay. 2. It was found to differ in several muscles. 3. That in external intercostal muscle, rectus abdominis muscle and splenius muscle from four horses was very high. 4. Although the masseter muscle had only type I fibers, CA-III content was similar to that in mixed-fiber type muscles such as the biceps femoris muscle. 5. It thus appear that equine type I fibers can be further subgrouped.
Richa , Grover YP, Charan S.The authors describe a rapid and simple dot immunobinding assay (DIA) for detection and identification of equine herpesvirus-1 antigen in field samples from cases of abortion, stillbirth, perinatal foal mortality and paralysis. The assay employs a nitrocellulose membrane to which antigen is adsorbed as a dot. Antigen is identified as a coloured dot using a procedure based on the principle of enzyme-linked immunosorbent assay (ELISA). In all, 61 samples were tested by DIA and the test was compared with conventional agar gel immunodiffusion (AGID). With DIA, 44 (72%) samples gave positive result...
Fikrig E, Magnarelli LA, Chen M, Anderson JF, Flavell RA.Enzyme-linked immunosorbent assays (ELISA) and immunoblots using either whole-cell lysates of Borrelia burgdorferi or an antigenic region of flagellin (41-G) as the antigen were performed, and the abilities of the two assays to detect antibodies to this spirochete in dog, cottontail rabbit, and horse sera were compared. Assays using whole-cell B. burgdorferi lysates as the antigen were more sensitive for detecting antibodies. ELISA with 41-G as the antigen were specific for Borrelia antibodies but were not as sensitive as the assays with whole-cell lysates coated to the solid phase. Use of rec...
Laegreid WW, Skowronek A, Stone-Marschat M, Burrage T.There are three clinicopathologic syndromes associated with African horsesickness (AHS) virus infection in horses. These different forms of AHS (pulmonary, cardiac, and fever forms) vary in the organs affected, the severity of lesions, time of onset of clinical signs and mortality rates. We have studied the effects of infection with three cell culture passaged variants of AHS virus in naive North American horses. One of these viruses, AHS/4SP, consistently caused the pulmonary form of AHS with rapid onset of severe pulmonary edema and 100% mortality. A second variant, AHS/9PI, resulted in sign...
Delbeke FT, Teale P, Debackere M, Houghton E.A commercially available generic promazine ELISA kit is available which shows cross-reactivity for the tranquilizer chlorprothixene (CPT). The ELISA test readily detects the presence of CPT or its metabolites in equine urine for up to 24 h after the i.v. and i.m. administration of sub-therapeutic doses (4.5 mg) to three horses. Maximum concentrations (CPT equivalents) are obtained 2 h after i.v. dosing. No distinct concentration peak values are observed after i.m. administration. Following solid-phase extraction, confirmation of CPT and its metabolites by electron impact mass spectrometry afte...
Afshar A, Shakarchi NH, Dulac GC.Two competitive (C) enzyme-linked immunosorbent assays (ELISAs) were developed for the detection of antibodies to vesicular stomatitis virus (VSV) in animal sera. The assays are based upon the availability of polyclonal antibodies (PAbs) from mouse ascitic fluids prepared against the New Jersey (NJ) and the Indiana (IN) VSV serotypes. The assays were performed by the immobilization of VSV-NJ and VSV-IN antigens on a solid phase (microtiter plate). Appropriately diluted test serum mixed with an equal volume of serotype-specific PAb was allowed to incubate in the presence of the relevant VSV ant...
Orino K, Saji M, Ozaki Y, Ohya T, Yamamoto S, Watanabe K.The effects of horse serum on the immunoassay of horse ferritin were investigated using two sandwich enzyme-linked immunosorbent assay (ELISA) systems. In System A, affinity-purified antibody to horse spleen ferritin and its conjugate with alkaline phosphatase were used as the first and second antibodies, respectively. In System B, whole antiserum and its conjugate with the enzyme were used. The recoveries of horse spleen ferritin added to horse sera were very low in either system (50-71% in System A; 42-79% in System B). However, heat treatment of the sera at 75 degrees C for 15 min improved ...
Lew AM, Thomas LM, Huntington PJ.Sera of sixteen horses with clinical signs of EIA from six different outbreaks and sera of 100 uninfected horses were used to validate an ELISA for EIA diagnosis. The antigen used was a recombinant protein derived from the amino-terminal portion of the transmembrane envelope protein of EIA (gp45). Reactivity between positive and negative sera could be clearly distinguished. Comparison with the traditional agar gel immunodiffusion test (commonly called the Coggins test) showed that the ELISA was superior in sensitivity. Comparison of this ELISA with the FAST-ELISA system showed that the latter ...
Corstvet RE, Gaunt SD, Karns PA, McBride JW, Battistini RA, Mauterer LA, Austin FW.An enzyme-linked immunosorbent assay (ELISA) was used to detect antigen in plasma and antibody in serum of 3 horses inoculated with Ehrlichia equi. Clinical signs, including rectal temperature, were correlated with the antigen and antibody detection. ELISA was very efficient in detection of serum antibody. Antigen detection using monoclonal antibodies to E. equi and ELISA should be considered as a diagnostic method.
Soulé C, Dupouy-Camet J, Georges P, Fontaine JJ, Ancelle T, Delvigne A, Perret C, Collobert C.Seven mares were infected with 20,000 Trichinella spiralis larvae; 2 of them were reinfected 22 wk later with the same amount of larvae. The course of infection in horses was assessed by serology (ELISA), biochemistry (aldolase activity), parasitology and histopathology. In each animal, infection was followed by a significant rise in specific antibody titers culminating at 5-10 wk post-infection (pi) and decreasing thereafter. Reinfection was followed by a slight rise in antibody levels. Aldolase activity increased during the first infection, but was not modified by reinfection. The parasite b...
Williams R, Du Plessis DH, Van Wyngaardt W.Group-reactive enzyme-linked immunosorbent assays (ELISAs) were developed to selectively detect antibodies to African horsesickness virus (AHSV) and equine encephalosis virus (EEV), 2 orbiviruses that infect equids. In indirect ELISA, guinea pig antisera to all known AHSV or EEV serotypes recognized immobilized AHSV serotype 3 or EEV Cascara, respectively. Antisera from naturally infected animals did not cross-react with their respective heterologous viruses. The ELISA was used in parallel with the complement fixation (CF) and agar gel immunodiffusion tests to detect antibodies in sera from an...
Pelegrino JL, Vázquez S, Morier L, Castillo A, Guzmán MG, Kourí G.We present the results attained in the identification of Eastern equine encephalomyelitis virus isolations in Vero and XL-2 cell systems, using a double-antibody ELISA technique and the indirect immunofluorescence method. The results attained through these two techniques coincided by 100% with identification through neutralization. With the former, the virus was detected within 6-8 hours after inoculation. Better results were attained with XL-2 cells.
Corstvet RE, Gaunt SD, Karns PA, Burgermeister D, McBride JW, Nicholson SM, Battistini RA.Four horses were inoculated with Ehrlichia risticii contained in either infected murine P388 D1 cells or heparinized blood from an infected horse. All 4 horses produced serum antibody, plasma antigen, and clinical signs of the disease. An enzyme-linked immunosorbent assay was used to detect antibody in the serum and was also used in conjunction with an anti-E. risticii monoclonal antibody to detect antigenemia. These laboratory and clinical findings were correlated to determine the efficiency of the antigen detection method for discerning E. risticii infection.
Qi T, Hu Y, Hu Z, Zhao S, Cullinane A, Lyons P, Gildea S, Wang X.Quantitation of virions is one of the important indexes in virological studies. To establish a sensitive and rapid quantitative detection method for equine arteritis virus (EAV), an antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) was developed by using two EAV nucleoprotein monoclonal antibodies (mAbs), 2B9 and 2B3, prepared in this study. After condition optimization, mAb 2B9 was used as the capture antibody, and HRP-labeled 2B3 was chosen as the detecting antibody. The AC-ELISA had a good standard curve when viral particles of the Bucyrus EAV strain were used as a reference stan...
Caloni F, Spotti M, Villa R, Mariani C, Montana M, Pompa G.An investigation was undertaken to demonstrate whether therapeutic treatment with ACTH raises hydrocortisone (cortisol) levels in horse urine above the limit (1000 ng/ml) established by the International Conference of Racing Authorities with the aim of controlling the abuse of cortisol and ACTH in equine sports. ACTH (200 iu) was administered i.m. to 3 Thoroughbred horses; urine and blood samples were collected at intervals afterwards and analysed by an immunoenzymatic system (ELISA) and HPLC-MS. To ascertain post exercise cortisol levels in untreated horses, 101 urine and 103 serum samples we...
Hall JA, Hoyt D, Zuver C, Skinner MM, Schlipf JW.The criteria used to diagnose recurrent airway obstruction (RAO) in affected horses include demonstration of reversible lower airway obstruction and greater than 25% neutrophils in bronchoalveolar lavage fluid (BALF). Additional objective laboratory tests are needed to improve diagnostic accuracy and to monitor response to treatment. The goal of this study was to determine if neutrophil chemoattractant activity of BALF could be measured by using a previously described, rapid, multiwell colorimetric assay for chemotaxis. In this assay, neutrophils that have migrated through a membrane filter ar...
Woods WE, Wang CJ, Houtz PK, Tai HH, Wood T, Weckman TJ, Yang JM, Chang SL, Blake JW, Tobin T.A one step enzyme-linked immunosorbent assay (ELISA) and a particle concentration fluorescent immunoassay (PCFIA) test for furosemide were evaluated as part of a panel of pre- and post-race tests for illegal medication of racing horses. These tests are very sensitive to furosemide with an I-50 for furosemide of about 20 ng/ml. The test is also rapid; an average pre-race complement of 10 samples can be analyzed in 90 minutes or less. The ELISA test results can be read with an inexpensive spectrophotometer, or even by eye. Both the PCFIA test and the ELISA test readily detect the presence of fur...
Mulville P.In the late 1970s, a new infectious disease in horses, involving acute enteritis, was recognised in the Potomac River area of Maryland, U.S.A. The causative agent was identified subsequently as a new species of rickettsial organism, later named Ehrlichia risticii. Since then, the disease has been reported in many other states, and in enzootic areas vaccination is common. Signs associated with the clinical disease included depression, fever, anorexia, decreased or absent intestinal sounds, profuse watery diarrhoea and laminitis. However, considerable variation in clinical manifestations has bee...
Barrey E, Valette JP, Jouglin M, Picard B, Geay Y, Robelin J.The content in slow and fast myosin heavy chains (MHC 1 and MHC 2) of 5 equine muscles was determined using an enzyme-linked immunosorbent assay. The results obtained with this immunoenzymatic method were compared with complementary techniques: electrophoresis and immunohistochemistry. Slices of masseter, diaphragm, tensor faciae latae, semitendinosus and cutaneus trunci were obtained from a 12-year-old saddle horse after slaughter. Muscular proteins were specifically extracted to be analysed by ELISA. The technique used 2 complimentary monoclonal antibodies (MAb). MAb 1 was prepared from a hu...
Arias MS, Piñeiro P, Hillyer GV, Francisco I, Cazapal-Monteiro CF, Suárez JL, Morrondo P, Sánchez-Andrade R, Paz-Silva A.The utility of an enzyme-linked immunosorbent assay to determine the sensitization against the trematode Fasciola hepatica in horses from an endemic area (NW Spain) was assessed. Blood samples were collected from 536 horses and tested against a 2.9-kDa recombinant surface protein (FhrAPS) to estimate the presence of IgG antibodies. Data were analysed regarding several intrinsic (age, gender and breed) and extrinsic factors (aptitude and housing). The farm size (number of horses/farm) was also considered. Sixty percent (95% CI 56, 64) of the horses were positive to the FhrAPS-ELISA, with a sign...
Schelp C, Böse R, Micha A, Hentrich B.High-titre equine immune sera were used to screen a lambda gt 11 expression library of Babesia equi cDNA fragments. Two cDNA clones which did not cross-hybridize to each other were studied. Both clones hybridized specifically to DNA from B. equi but not to DNA from B. caballi, B. divergens or B. ovis. Recombinant proteins were expressed as glutathione S-transferase (GST) fusion proteins with apparent molecular weights of 40 kDa and 75 kDa. Polyclonal antibodies directed against the 40 kDa and 75 kDa recombinant proteins detected native antigens of 55 kDa and 50 kDa respectively in crude lysate...
Brown MP, Trumble TN, Merritt KA.To investigate effects of osteochondral injury on high-mobility group box chromosomal protein 1 (HMGB-1) concentrations in synovial fluid (SF) from Thoroughbreds and to compare these results with radiographic and arthroscopic scores of severity of joint injury. Methods: 40 clinically normal rested Thoroughbreds (group 1) and 45 Thoroughbreds with osteochondral injury as a result of racing. Methods: SF was obtained from the metacarpophalangeal (MCP) joints, metatarsophalangeal (MTP) joints, middle carpal joints, and radiocarpal joints. For group 2, radiographic and arthroscopic scores were dete...
Dirikolu L, Mollett BA, Troppmann A, Woods WE, Bratton C, Cashman CP, Schroedter D, Mayer B, Lehner AF, Karpiesiuk W, Hughes C, Boyles J, Harkins JD....Single doses of one, three, and six actuations (120 micro g albuterol/actuation) and multiple daily doses (six actuations per dose four times daily) for 5 days of aerosol albuterol sulfate were sequentially administered to each of six horses using an equine inhaler device (Torpex, 3M Animal Care Products, St. Paul, MN [corrected] and Boehringer Ingleheim Vetmedica, Inc., St. Joseph, MO [corrected]). A 2-week washout period was allowed between each dose. ELISA testing revealed no evidence of albuterol in urine at 24 hours after any single-dose administration. Results indicated that 48 hours or ...
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...
O'Brien MA, Holmes MA, Duffus WP.Anti-tetanus toxoid (TT) antibody (Ig) levels in the supernatant of cultured, pre-immunised equine peripheral blood mononuclear cells (PBMC) were measured by an indirect enzyme-linked immunoabsorbent assay (ELISA). Optimal anti-TT Ig production occurred at concentrations of stimulating, purified TT of between 0.001 and 0.1 micrograms ml-1, which varied depending on the cell concentration. Optimal anti-TT Ig production was most consistently produced when the cell concentration was 5 x 10(6) ml-1. At this cell concentration maximal anti-TT Ig was induced using 0.1 micrograms ml-1 TT. At a cell c...
Smith JM, Meyers KM, Barbee DD, Schott H, Bayly WM.To determine whether plasma von Willebrand factor (vWf) concentration changes in horses during and after treadmill exercise. Methods: 5 mature, fit Thoroughbreds. Methods: A blood sampling catheter was placed in the right jugular vein. A warm-up period was followed by a 3-minute rest period. Horses were galloped at racing pace until fatigued (about 2 minutes). Blood samples were collected prior to warm-up, during the postwarm-up rest period, 1 minute into the run, at cessation of the run, and 5 to 120 minutes after cessation of the run. vWf activity was measured by ELISA and corrected for plas...
Matheson A, Regmi SC, Jay GD, Schmidt TA, Scott WM. Local biological and biomechanical-stimuli modulate proteoglycan-4 secretion within synovial joints. For the horse, changes to proteoglycan-4 concentration and function are notable in acute joint injury and osteoarthritis. Proteoglycan-4 (also known as Lubricin) is present in the blood, however the effect of exercise on equine serum levels is unknown. The overall objective of this study was, therefore, to investigate the effect of intense exercise on serum proteoglycan-4 in thoroughbred horses. Samples of blood were taken from thoroughbreds ( = 12) during a chuckwagon racing event (Alberta, ...
Fu ZF, Denby L, Lien DH, Robinson AJ.An indirect enzyme-linked immunosorbent assay (ELISA) was developed for the detection of antibodies against equine herpesvirus type 2 (EHV-2) in equine sera. The optimal conditions of antigen concentration, and serum and conjugate dilutions were established by chequerboard titrations. When the standard ELISA test was used for titration of test sera, it was found to give titres approximately 1500 times higher than those obtained in the virus neutralization (VN) test, and a correlation coefficient of 0.815 was obtained between these two tests on 42 equine sera. All the positive serum samples by ...
Palme R, Entenfellner U, Hoi H, Möstl E.Non-invasive pregnancy diagnosis in mares by measuring faecal oestrogens has been performed over years with great accuracy. However, results have indicated breed-related differences in the amount of excreted steroids during late pregnancy. Therefore faecal samples were collected during the last 4 months of pregnancy of Thoroughbred (n = 10), New Forest pony (n = 9), Shetland pony (n = 10) and Iceland pony mares (n = 11). Concentrations of oestrogens, 20alpha-hydroxy- and 20-oxopregnanes were measured using enzyme immunoassays. Breed differences concerning both levels (though significant only i...
de Arruda Coutinho LC, de Jesus AL, de Paiva Fontes KF, Coimbra EC, Mariz FC, de Freitas AC, de Cássia Carvalho Maia R, de Castro RS.Equine Infectious Anemia (EIA) is a persistent lentivirus infection of horses which causes a chronic clinical condition with worldwide importance in veterinary medicine. The p26 protein is usually prepared for use as an antigen in serological tests for EIA diagnosis since it is a well-conserved gene sequence and very immunogenic. In view of the ability of yeast to make post-translational modifications of proteins, this study was carried out to allow Pichia pastoris to be used for the expression of a synthetic codon-optimized EIAV p26 gene. The gene was cloned into pPICZαA vector after appropr...
Kranenburg LC, van Ree HE, Calis AN, de Pater M, Buter GJ, van Maanen C, Sloet van Oldruitenborgh-Oosterbaarn MM.Equine proliferative enteropathy caused by Lawsonia intracellularis is an emerging disease of weanling foals and affects their growth and development. The prevalence of Lawsonia intracellularis in The Netherlands is not known. The aim of the study was to investigate the seroprevalence of Lawsonia intracellularis in horses in The Netherlands. Blood samples were taken from healthy foals before and after weaning and from healthy yearlings and mature horses on farms throughout The Netherlands. These samples were analysed for the presence of Lawsonia intracellularis-specific antibodies with a block...
Kaiser A, Meier HP, Doherr MG, Perler L, Zanoni R, Gerber V.Since 1991, no cases of Equine Infectious Anemia (EIA) have been reported in Switzerland. Risk factors for introduction of the virus into Switzerland are still present or have even increased as frequent inapparent infections, large numbers of imported horses, (since 2003) absence of compulsory testing prior to importation, EIA cases in surrounding Europe, possible illegal importation of horses, frequent short-term stays, poor knowledge of the disease among horse owners and even veterinarians. The aim of this study was to provide evidence of freedom from EIA in imported and domestic horses in S...
Drummer HE, Reynolds A, Studdert MJ, MacPherson CM, Crabb BS.Sera from 33 Australian thoroughbred mares were tested during an outbreak of equine herpesvirus 1 (EHV1) abortion with an enzyme-linked immunosorbant assay (ELISA) for the presence of EHV1-specific antibodies. The ELISA used a recombinant EHV1 antigen derived from glycoprotein G (gG) and distinguished antibodies to EHV1 from those of the antigenically related and widespread herpesvirus EHV4. Sera were obtained from most of the mares on three occasions, three, 13 and 67 days after the first abortion. Mares which were negative in the ELISA were kept separate from mares which were positive. A sec...
Topper MJ, Prasse KW, Morris MJ, Duncan A, Crowe NA.To adapt and characterize a human ELISA kit to quantify thrombin-antithrombin III (TAT) complexes in horses, and to evaluate TAT as a marker for hypercoagulation in horses. Methods: 29 clinically normal horses used as controls, and 4 ill horses used to evaluate assay for known causes of hypercoagulation. Methods: A commercially available human sandwich-type ELISA kit with 2 antibodies against human thrombin and antithrombin III that bind selectively to their corresponding TAT antigenic sites was used. Equine TAT standards were made from purified equine thrombin and antithrombin III. Proteins d...
Bidwell LA, Wilson DV, Caron JP.To measure concentrations of lidocaine serum after application of two 5% patches on horses. Methods: Prospective experimental trial Methods: Six client-owned, systemically healthy horses. Methods: The hair was clipped on the medial aspect above the carpus of both fore limbs and 2 patches of 5% lidocaine were applied within 30 minutes of jugular catheter placement and the area was then bandaged. Venous blood was drawn from a jugular vein catheter that was inserted using lidocaine as a local block. Samples were drawn immediately before and at 2, 4, 6, 8, and 12 hours after patch application. The...
Keating DL, Lehman JS, Burk SV.Identification of factors associated with parasite infestation in horses could reduce frequency of anthelmintic administration and slow development of anthelmintic resistance. The study aim was to evaluate management factors, equine characteristics, and equine salivary cortisol concentrations for association with strongyle-type egg shedding levels. As immunocompromised horses appear to be more susceptible to parasite infestation, it was hypothesized that salivary cortisol concentration could be associated with parasite egg shedding. Saliva and fecal samples were collected from 200 horses acros...
Fernandes I, Lima EX, Takehara HA, Moura-da-Silva AM, Tanjoni I, Gutiérrez JM.Horse IgG isotypes and cross-neutralization of two snake antivenoms produced in Brazil and Costa Rica. Toxicon 000-000. This work compared the specificity, ELISA titers and IgG subclass content of the polyvalent antivenom (anti-Bothrops asper, Crotalus durissus durissus and Lachesis muta stenophrys) of Instituto Clodomiro Picado (Costa Rica) and the bothropic antivenom (anti-Bothrops jararaca, B. jararacussu, B. moojeni, B. neuwiedi and B. alternatus) of Instituto Butantan (Brazil). The role of IgG(T) and IgGa subclasses in neutralization of some venom toxic activities and the cross neutraliza...
Scudamore CL, Pemberton AD, Miller HR, McDonnell AM, Thomson SR, Dawson A, Watson ED.An enzyme linked immunosorbent assay (ELISA) was developed and used to estimate the concentrations of the serine proteinase inhibitor, alpha-1 proteinase inhibitor (API), in uterine flushings recovered from mares at different stages of the oestrous cycle and before and after the induction of experimental endometritis. There was a significant increase in the concentrations of API and albumin relative to total protein in flushings recovered during oestrus compared with dioestrus but no difference was observed in the concentrations of these proteins relative to total protein before and after the ...
Hu WG, Alvi AZ, Fulton RE, Suresh MR, Nagata LP.A recombinant gene encoding a single-chain variable fragment (scFv) antibody against Venezuelan equine encephalitis virus (VEE) was cloned into a prokaryotic T7 RNA polymerase-regulated expression vector. A streptavidin-binding peptide gene fused to a 6His tag was attached downstream to the scFv gene. The recombinant fusion protein was expressed in bacteria as inclusion bodies that were subsequently solubilized with 8 M urea and renatured by an arginine system. Purification of the fusion protein was achieved by immobilized metal affinity chromatography. Enzyme-linked immunosorbent assay (ELISA...
Chen J, Guo X, Li L.The nucleocapsid (N) protein is the most conserved structural protein in equine arteritis virus (EAV). This study aimed to identify the minimal conserved B cell epitope on the EAV N protein. The purified N protein was used to immunize mice for preparing monoclonal antibody (mAb). The reactivity of mAb was evaluated by Western blot and immunofluorescence assay. Moreover, 11 overlapping peptides (named MBP-N1 to MBP-N11) were designed to localize the linear antigenic epitope within the N protein. The peptides were identified by indirect enzyme-linked immunosorbent assay (ELISA) and Western blot....
Dirikolu L, Lehner AF, Harkins JD, Woods WE, Karpiesiuk W, Gates RS, Fisher M, Tobin T.Pyrilamine is an antihistamine used in human and veterinary medicine. As antihistamines produce central nervous system effects in horses, pyrilamine has the potential to affect the performance of racehorses. In the present study, O-desmethylpyrilamine (O-DMP) was observed to be the predominant equine urinary metabolite of pyrilamine. After intravenous (i.v.) administration of pyrilamine (300 mg/horse), serum pyrilamine concentrations declined from about 280 ng/mL at 5 min postdose to about 2.5 ng/mL at 8 h postdose. After oral administration of pyrilamine (300 mg/horse), serum concentrations p...
Tobin T, Kwiatkowski S, Watt DS, Tai HH, Tai CL, Woods WE, Goodman JP, Taylor DG, Weckman TJ, Yang JM.We have developed and evaluated a one step enzyme-linked immunosorbent assay (ELISA) test for sufentanil and a 125I radioimmunoassay test for alfentanil as part of a panel of pre- and post-race tests for narcotic analgesics in racing horses. Our sufentanil ELISA test detects sufentanil with an I-50 of about 0.5 ng/ml. The test is rapid and economical in that it can be read with an inexpensive spectrophotometer, or even by eye. The test readily detects the presence of sufentanil or its metabolites in equine blood and urine from 1 to 24 hours respectively after administration of therapeutic or s...
