Immunoassays are analytical techniques used to detect and quantify specific proteins, hormones, or other molecules in biological samples from horses. These assays rely on the specificity of antibodies to bind to target antigens, allowing for the measurement of various substances that can indicate health status or disease. In equine research, immunoassays are employed to assess biomarkers related to metabolic, infectious, and inflammatory conditions. Common applications include measuring hormone levels, detecting infectious agents, and monitoring immune responses. This page compiles peer-reviewed research studies and scholarly articles that explore the development, application, and validation of immunoassays in the context of equine health.
Yamamoto K, Hashimoto K, Chiba J, Simizu B.To analyze the biological activities of the alphavirus glycoproteins, eight different monoclonal antibodies against the two glycoproteins of western equine encephalitis virus were isolated. Five of the eight monoclonal antibodies were shown to be specific for E1 and three for E2 protein by an enzyme-linked immunosorbent assay and by radioimmunoprecipitation. Three of the five anti-E1 and all of the anti-E2 monoclonal antibodies inhibited hemagglutination by purified virions. One anti-E1 and two anti-E2 monoclonal antibodies possessed high virus-neutralizing activity.
Haddad NS, Pedersoli WM, Ravis WR, Fazeli MH, Carson RL.Gentamicin (GT) was administered IM to 6 healthy mature mare ponies at a dosage of 5 mg/kg of body weight every 8 hours for 7 consecutive days (total, 21 doses). Two venous blood samples were collected before (trough) and at 1 hour (peak) after the 5th, 10th, 14th, and 19th doses. An endometrial biopsy was done of each mare on days 4 and 7. On the 7th day, just before the 21st administration of GT, base-line blood samples were collected, and 22 blood samples were collected over a period of 48 hours after GT was given. The mares were catheterized on the 7th day, and urine was collected for 24 h...
Galan JE, Timoney JF.Mucosal nasopharyngeal immunoglobulin A (IgA) and IgG responses to proteins of Streptococcus equi were studied in horses after the experimental production of strangles. S. equi-specific IgA and IgG titers in nasopharyngeal mucus were much higher in samples from animals 1 to 2 weeks after challenge than in samples from control animals. Although IgA was the major immunoglobulin in nasal mucus, there was more antibody activity associated with IgG as measured by radioimmunoassay. Great differences between the specificities of antibodies in nasal mucus and in serum were detected. IgA and IgG of muc...
Benoit E, Garnier F, Courtot D, Delatour P.Antiserum has been raised in rabbits treated with a 19 nor testosterone-hemisuccinate-bovine-serum-albumin conjugate and used for the development of a specific RIA of plasma 19 nor testosterone. Plasma samples are drawn from testicular and jugular veins of stallions during castration under general anesthesia. Results demonstrate a testicular secretion of 19 nor testosterone and a stress inhibition of this secretion correlatively with stress inhibition of testosterone secretion.
Bottoms GD, Johnson MA, Lamar CH, Fessler JF, Turek JJ.Dispersed equine vascular endothelial cells grown in tissue culture, and freshly isolated neutrophils were used to determine direct effects of endotoxin on cyclooxygenase and lipoxygenase products. Endothelial cells (10(7)/ml) or neutrophils (2 X 10(6)/ml) were incubated with (a) buffer, (b) endotoxin (10 micrograms/ml), (c) endotoxin + flunixin meglumine (10 micrograms/ml), or (d) calcium ionophore, A23187 (10 micrograms/ml). Thromboxane (TxB2), prostacyclin (6-keto-PGF1 alpha), and leukotriene C4 (LTC4) were determined in the incubation fluid by radioimmunoassay. Thromboxane and prostacyclin...
Glaze MB, McGuire TC, Schmidt GM, Leid RW.A quantitative investigation of equine tear and aqueous humor immunoglobulins was done using normal horses and ponies as well as horses and ponies infected with Onchocerca cervicalis. The equine immunoglobulin isotypes IgGa, IgM, IgA and IgG(T) were quantitated by either single radial immunodiffusion (SRID) or radioimmunoassay (RIA). Tear immunoglobulin levels for IgGa (128 +/- 151 micrograms/ml), IgA (1,664 +/- 1,038 micrograms/ml) and IgM (106 +/- 74 micrograms/ml) were measured, while IgG(T) was not detectable. In horses with ocular inflammation the IgGa was 18-fold higher in the tears, 2,2...
Bishop AE, Hodson NP, Major JH, Probert L, Yeats J, Edwards GB, Wright JA, Bloom SR, Polak JM.In recent years, distinct changes in regulatory peptides have been found in a number of gastrointestinal diseases. Grass sickness is a fatal disease of horses for which the etiology has yet to be fully ascertained. In this study, the peptide-containing nerves and ganglionic and mucosal endocrine cells of the ileum, colon and rectum were investigated in horses with sub-acute or chronic grass sickness and compared with normal controls using immunocytochemistry, at both the light and electron microscopical levels, and radioimmunoassay. A substantial loss of both peptide-containing cells and nerve...
Montelaro RC, West M, Issel CJ.The immunogenic contributions of the carbohydrate and peptide portions of the major envelope glycoprotein of equine infections anemia virus, EIAV gp90, were analyzed by measuring the effects of specific glycosidase and protease digestions on the reactivity of the glycoprotein with immune sera from infected horses. The results of both direct and competitive radioimmunoassay demonstrated that immune sera contained antibodies reactive with both the carbohydrate and protein moieties of EIAV gp90, with the predominant reactivity apparently against the gp90 peptide epitopes. These results contrast w...
Silberzahn P, Zwain I, Martin B.Blood testosterone levels were measured by RIA and gas chromatography-mass spectrometry in the pregnant mare. They were found to increase from the very beginning of pregnancy, reaching peak values 10 times higher than the basal values at the seventh month and then to return to basal values by the week after parturition. Testosterone binding by plasma proteins was investigated in nonpregnant and pregnant mares throughout gestation. Equilibrium dialysis and gel equilibration methods did not reveal any blood specific testosterone-binding activity at any gestational stage. Hence, blood testosteron...
Higgins AJ, Lees P, Taylor JB.In a two part cross-over experiment, acute inflammatory exudates were induced in 7 ponies by subcutaneous implantation of 3 sterile carrageenin-soaked polyester sponge strips. Treatment comprised a single therapeutic geenin-soaked polyester sponge strips. Treatment comprised a single therapeutic dose of 4.4 mg/kg phenylbutazone (PBZ) administered intravenously at the time of sponge implantation. Exudates were harvested at 6, 12 and 24 hours and examined for leukocyte and erythrocyte numbers using the improved Neubauer technique; for eicosanoids by radioimmunoassay and by high performance liqui...
Hart PJ, Squires EL, Imel KJ, Nett TM.Seasonal changes in the hypothalamic-hypophyseal axis were investigated using tissue from 49 light-horse mares, of mixed breeding. Hypothalamic and pituitary tissues were collected at 5 intervals throughout the years 1981 and 1982, representing midbreeding season (July, n = 10), transition out of the breeding season (October, n = 11), midanestrus (December, n = 8), transition into the breeding season (March, n = 10), and again in the following midbreeding season (July, n = 10). The hypothalamic region was dissected into preoptic area, body and median eminence. Gonadotropin-releasing hormone (G...
Wichmann U, Wichmann G, Krause W.Blood levels of testosterone precursors, i.e. pregnenolone, progesterone, 17 alpha-hydroxyprogesterone, androstendione, DHEA, and delta 5-androstendiol as well as testosterone and estradiol are measured in 10 animals each of 10 different species. The determination is done by radioimmunoassay with steroidspecific antibodies. Precursors of the delta 5-pathway (DHEA, androstendiol) are low in the red deer, dog, cat, rat and guinea pig. Precursors of the delta 4-pathway (progesterone, 17-hydroxprogesterone, androstendione) are lower in the bull, boar, ram, stallion and rabbit thus indicating a pre...
Roser JF, Chang YS, Papkoff H, Li CH.A specific and sensitive homologous radioimmunoassay has been developed for equine prolactin, suitable for measuring prolactin concentrations in serum of horses. The sensitivity of the assay ranged from 0.4 to 0.6 ng/ml and the intra- and inter-assay coefficients of variation averaged 6.9 and 15.4%, respectively, for five doses of hormone. Cross-reactivity with other mammalian and nonmammalian prolactins and growth hormones was less than 20 and 0.3%, respectively. Cross-reactivity with equine growth hormone was less than 0.07%. Equine serum and pituitary extracts showed parallel dilution-respo...
Higgins AJ, Lees P.The presence of cyclooxygenase products of arachidonic acid metabolism in carrageenin-induced inflammatory exudate was investigated in ponies using two models. In the first model, an inflammatory response was stimulated by injecting carrageenin into subcutaneously implanted polypropylene tissue cages and exudates were collected at five predetermined times between 3 and 48 h. In the second model, exudates were harvested at 6, 12 and 24 h from carrageenin-impregnated polyester sponges which had also been inserted beneath the skin. Prostaglandin (PG) E2, thromboxane (TX) B2 and the stable breakdo...
Gibson SJ, Polak JM, Anand P, Blank MA, Morrison JF, Kelly JS, Bloom SR.The distribution of VIP-immunoreactivity was studied in the spinal cord and dorsal root ganglia of 6 mammalian species. Immunoreactive fibres and cell bodies were most apparent in the dorsal horn, dorsolateral funiculus, intermediolateral cell columns and the area around the central canal. The distribution of VIP immunoreactivity was similar in all species studied, mouse, rat, guinea pig, cat, horse and the marmoset monkey. There were fewer VIP fibres in the dorsal horn of cervical and thoracic segments than in lumbosacral segments. Using radioimmunoassay this gradient increase was quantitativ...
Altmeyer P, Holzmann H, Stöhr L, Koch HJ.White horses are subject to age-dependent coat depigmentation. They are dark gray or black at birth and lose their coloring between their second and fourth year. Beginning at about age 10 their coat takes on a characteristic silver-gray coloring. The purpose of this paper was to find out to what extent the endogenic alpha-MSH level changes with the change in pigmentation. alpha-MSH plasma levels were determined by radioimmunologic analysis in 3 age groups of white Camarque horses: age group 1 consisted of dark horses with a mean age of 1.2 years and a mean alpha-MSH level of 106.4 pg/ml +/- 18...
Urwin V.Heterologous double-antibody radioimmunoassay were developed for the measurement of FSH and LH concentrations in the serum of both horses and donkeys. The FSH assay employed a rabbit anti-ovine FSH serum which showed a complete lack of cross-reaction with equine chorionic gonadotrophin (eCG) and negligible cross-reaction with equine LH. The LH assay utilized an antiserum raised against highly purified eCG. This similarly showed negligible cross-reaction with equine FSH but its high cross-reactivity with eCG prevented the measurement of equine LH concentrations in serum when eCG was also presen...
Bossut DF, Leshin LS, Malven PV.Radioimmunoassay procedures were developed and validated for the quantification of beta-endorphin (beta-EP)-like immunoreactivity in equine plasma. beta-EP could be quantitatively extracted from plasma with silicic acid powder and subsequently assayed, however, valid estimates of this hormone could also be obtained on unextracted plasma. Although beta-lipotropin (beta-LPH) cross-reacted in the assay, it was not necessary to correct for beta-LPH activity when assaying unextracted plasma because chromatographic analyses showed that 92% of the immunoreactivity in plasma extracts was similar in mo...
Makawiti DW, Allen WE, Kilpatrick MJ.A simple and rapid (less than 2 h) immunoassay method has been developed based upon a novel separation technique called LIDIA (Ligand Differentiation Immunoassay), enabling direct estimation of the concentration of oestrone sulphate in ethanolic extracts of blood plasma. An antiserum raised against oestrone-3-glucuronyl-BSA was used which showed a higher cross-reaction with the sulphate than the glucuronide metabolite. The assay had a sensitivity of 5.2 pg/tube and acceptable inter-(less than 18%) and intra-(less than 8.5%) assay precision. Analysis of samples of peripheral venous plasma obtai...
van Leeuwen W, Noden PA, Dieleman SJ.Three experiments were conducted to test the abortifacient effects of PGF2 alpha analogues on mares during midgestation (average gestation length 141.5 days). The progesterone concentration was measured by radioimmunoassay. In experiment 1. five mares received an injection of PGF2 alpha analogue (fluprostenol: 500 micrograms intramuscularly) and a second injection either at 24, 48, of 72 h. Although the progesterone concentration decreased (P less than 0.05) an average of 44 per cent in 24 h, none of the pregnancies were terminated. In experiment 2, beginning at least 10 days after experiment ...
Thompson DL, Reville SI, Walker MP, Derrick DJ, Papkoff H.To study the possible role of ovarian androgens in regulation of follicle stimulating hormone (FSH) secretion in the cycling mare, five mature, intact mares were treated with testosterone (20 micrograms/kg of body weight) daily during estrus; five control mares received safflower oil on the same schedule. Mares were teased for estrus and samples of jugular blood were drawn daily through one full estrous cycle. Concentrations of FSH in plasma were measured by a newly developed radioimmunoassay based on anti-ovine FSH serum and radioiodinated equine FSH. Testosterone treatment during estrus had ...
Silberzahn P, Dehennin L, Zwaïn IH, Leymarie P.Testosterone has been identified by mass spectrometry in blood and follicular fluid aspirated from mature Graafian follicles of mares. Quantitative measurements made by gas chromatography-mass spectrometry have validated the determination of plasma testosterone made by radioimmunoassay. However, because of high levels of epitestosterone (17 alpha-hydroxyandrost-4-en-3-one) in the follicular fluid, radioimmunoassay overestimates the true concentrations of testosterone. The occurrence of testosterone in mare follicular fluid at a concentration which is two orders of magnitude higher than that in...
Roberts MC.Vitamin B12 and folate concentrations were determined by radioimmunoassay in groups of horses in Queensland. Highest serum vitamin B12 levels were found in supplemented performance horses. These, together with pastured horses that included pregnant and lactating mares, had significantly greater serum folate activity than permanently stabled animals. The range of red cell folate concentrations was much narrower in horses in training than from any other group. Red cell folate may be a better indicator of a horse's folate status than the serum folate value. Vitamin B12 and folate concentrations w...
Tamanini C, Giordano N, Chiesa F, Seren E.Plasma cortisol variations have been determined by radioimmunoassay in 5 stallions during mating and in 2 teasers during oestrous female exposure. In all the animals, cortisol plasma levels consistently increase (71.1 ng/ml vs 44.0 and 63.0 ng/ml vs 35.1, in the stallions and in the teasers, respectively) 7-30 min after female exposure; 120 min after exposure, cortisol concentrations are again low.
Morris DD, Garcia M.Adult horses showed a mild diurnal variation in equine plasma thyroxine (T4) concentrations, but not triiodothyronine (T3). Plasma T4 concentrations tended to be higher between 5 PM and 8 PM than at 8 AM. Increases in plasma T4 and T3 were similar in adult healthy horses given 5, 10, or 20 IU of thyroid-stimulating hormone (TSH). The T4 peaked at approximately twice (2.0 +/- 0.4 times) as high as the base line at 6 to 12 hours after the TSH was given. The greatest change from base line T3 occurred at 1 to 3 hours after the TSH was given, but the magnitude of increase was widely variable (4.36 ...
Montelaro RC, West M, Ivey M.Representative glycoproteins including fetuin, protein A, ovalbumin, alpha 1 acid glycoprotein, and the major glycoprotein of equine infectious anemia virus were labelled with 125I by the chloramine-T or Bolton-Hunter procedure and their binding to immobilized Con A or lentil lectin compared to untreated samples of each glycoprotein. Glycoprotein modification was no greater than one substituted residue per protein molecule. Yet the radioiodinated glycoproteins typically displayed only 0-50% of the lectin binding observed with untreated samples. These results indicate that lectin glycoprotein b...
Latif ZA, Fletcher MA.Immunochemical comparisons were made of the reactivity of membrane glycoproteins from horse, bovine, sheep and goat erythrocytes with heterophile antibodies of infectious mononucleosis. The four receptors were tested as competitive inhibitors of a sandwich-type solid-phase radioimmunoassay and of agglutination of glycoprotein-latex reagents by infectious mononucleosis serum. The results of this study showed that the bovine glycoprotein had a superior reactivity with this heterophile antibody system and sheep erythrocyte glycoprotein was the least reactive. The latter had negligible ability to ...
Harkiss GD, Brown DL, Smith DJ, Nagington J.Circulating immune complexes were isolated from the sera of cardiac allograft recipients by bovine conglutinin/anti-conglutinin co-precipitation, or by gel filtration and protein A-Sepharose affinity chromatography. The antibody moieties within these isolated immune complexes were tested for specificity against heterologous anti-thymocyte globulins by solid phase radioimmunoassay, and bacterial and viral antigens by indirect immunofluorescence. The results showed that in addition to possessing specific anti-equine anti-thymocyte globulin antibodies, immune complexes also contained cross-reacti...
Thompson DL, Reville SI, Derrick DJ.Five mature Quarterhorse mares were bled every 30 min for 25 h on day 50 of pregnancy to determine the short-term mode of secretion of equine chorionic gonadotropin (eCG). Three other mares with persistent endometrial cups after abortion were administered gonadotropin releasing hormone (GnRH; 1.0 mug/kg of body weight) and were bled immediately prior to and at 15, 30, 45, 60, 90, 120, 180 and 240 min after GnRH. Concentrations of eCG in plasma of pregnant mares were constant over the 24-h period; the variation of each mare's individual values was no greater (P>.05) than the predicted random...
McKeever KH, Hinchcliff KW, Cooley JL, Lamb DR, Muir WW.Six nontrained mares were subjected to steady-state, submaximal treadmill exercise to examine the effect of exercise on the plasma concentration of atrial natriuretic peptide (ANP) in arterial, compared with mixed venous, blood. Horses ran on a treadmill up a 6 degree grade for 20 minutes at a speed calculated to require a power equivalent to 80% of maximal oxygen uptake (VO2MAX). Arterial and mixed venous blood samples were collected simultaneously from the carotid and pulmonary arteries of horses at rest and at 10 and 20 minutes of exercise. Plasma was stored at -80 C and was later thawed; A...
Janowski T, Skolimowska A, Zdunczyk S, Baranski W.Total of 30 pregnant mares were used in this study: 17 Thoroughbred and 13 Primitive Polish Horse. The concentration of total unconjugated oestrogens was determined by radioimmunoassay in faecal samples obtained every second week after mating until delivery or interruption of pregnancy. Concentration of oestrogens in pregnant healthy mares ranged from 40 to 200 ng/g and showed typical profiles for pregnancy. In 5 of 30 mares (16.6%) of either breed abortions and cases of embryonic death occurred. Abortions were preceded by sharp or gradual decrease in oestrogen concentration, whereas prior to ...
Secor EJ, Womack SJ, Ysebaert MP, Colville MJ, Reesink HL.Synovial fluid (SF) is an ideal sentinel fluid for osteoarthritis (OA) diagnosis and prognostication due to its critical homeostatic role, proximity to articular tissues and immune cell composition. Untargeted proteomics enable identification of soluble markers for diagnostic and therapeutic applications while minimising bias. Objective: To use liquid chromatography-tandem mass spectrometry (LC-MS/MS) to define the SF proteome in horses with and without carpal OA. The goal was to identify differentially regulated proteins in mild-moderate carpal joint disease compared with healthy joints. Meth...
Otto P, Elschner M, Schulze P, Prudlo J, Schrader R.The commercially available immunoassay "OnSite Rotavirus" was used for the detection of animal rotaviruses in 113 faecal samples. The sensitivity of the test was 88% and the specificity 96% compared with reference methods (EIA, EM). This test would detect approximately 4.4 x 10(6) to 1.8 x 10(7) virus particles per ml. The presence of virus could be demonstrated in fresh faecal samples from cattle, horses and pigs within a few minutes. The rotaviruses of group A were identified independently of the virus serotype. Further results and additional problems of using this test kit are described.
Allen AL, Scott WM, Cook SJ, Fretz PB, Doige CE.Studies were conducted to determine the effects of delaying the separation of serum from the clot and of long-term storage of serum samples on the measurement of thyroid hormones in blood from horses using a fluorescence polarization immunoassay. The measured concentrations of T3 and T4 were not affected by leaving serum on the clot for as long as 24 hours at room temperatures. Storage of serum for 19 to 22 months at -20 degrees C resulted in significant increases of measured T4, but not T3. These studies support previous work demonstrating that thyroid hormones are resistant to degradation, i...
McKellar QA, Nolan AM, Galbraith EA.Blood collected from calves, sheep, goats, pigs, dogs, horses, ponies and donkeys, was allowed to clot under standard conditions. Thromboxane B2 generated during the clotting process was measured by radioimmunoassay in serum harvested from each sample. Highly significant differences were found between species and also between genera within a species. Highest concentrations of thromboxane B2 were detected in the dog samples (887.7 +/- 123.7 ng/ml) and lowest concentrations in samples from sheep (2.7 +/- 0.2 ng/ml). The amount of thromboxane produced per unit number of circulating platelets or p...
Hafs HD.Often in developing hormone assays, hormones that may interfere with the assay by cross-reaction are not available for testing the validity of the assay. For example, horse TSH was unavailable to test for cross-reaction in an LH radioimmunoassay (RIA). The authors devised an indirect means of accomplishing the same goal, and the evidence from the indirect test of cross-reaction was at least as persuasive as a direct test might have been. Other examples are given of experiments where extensive effort was devoted to validation of steroid RIA, but there were substantial quantitative differences i...
Lindner A, Will Y, Chrispeels J.Reference values for Cortisol, T4 and T-Uptake, determined with the Fluorescence-Polarization-Immunoassays (FPIAs) in blood-plasma of different horse groups were established. The highest Cortisol values were measured in blood samples from thoroughbred racehorses and riding horses taken between 7 and 8 a.m. (181 +/- 37 and 268 +/- 43 nmol/l), the lowest gained between 5 and 6 p.m. (69 +/- 45 and 85 +/- 32 nmol/l respectively). Peak values for T4 in riding horses were found in blood samples collected between 1 and 2 p.m. (28.2 +/- 5.7 nmol/l) followed by the samples taken at 5-6 p.m. and 7-8 a.m...
Kowalik S, Kedzierski W.The effect of interval vs. continuous exercise on plasma leptin and ghrelin concentration in young Standardbred horses was studied. The experiment was conducted on 27 trotters, in the age between 2 and 3 years. They were divided into two groups according to the type of exercise. Blood samples were collected through jugular venipuncture in the following experimental conditions: at rest, immediately after exercise and 30 minutes after the end of the effort. Plasma leptin and ghrelin concentrations were determined using RIA tests. The continuous exercise induced an increase in plasma leptin conce...
King SS, Evans JW.Follicular growth, circulating estradiol concentrations and endometrial prostaglandin F2 alpha (PGF2 alpha) production were measured to determine whether there is an interrelationship among these factors associated with luteolysis. Follicular growth was monitored by rectal palpation every other day during diestrus in 16 mares. Plasma estradiol was determined for daily samples during all estrous cycles. Endometrial tissue was removed for PGF2 alpha analysis by radioimmunoassay on d 10, 12, 14 or 16 during several normal cycles and after d 30 during spontaneously prolonged cycles. Circulating es...
Chapman DI, Whiteside J.A radioimmunoassay method has been developed that enables the administration of therapeutic doses of synthetic corticosteroids to be detected in horse urine. Fourteen proprietary preparations of these steroids have been given by intramuscular injection to ponies and thoroughbreds. The administration of some preperations could still be detected six days after a single intramuscular injection of a therapeutic dose. The route of injection of dexamethasone-21-sodium phosphate, whether intramuscular, intravenous or intra-articular, did not appear to alter the length of time over which the steroid o...
Rearden TP, Sprouse RF, Garner HE.A radioimmunoassay was developed to discriminate immunoglobulin (Ig) classes specific for the J-5 mutant of Escherichia coli (serotype O:111-B4). Adult horses were periodically inoculated IM with a nonviable suspension of the J-5 mutant emulsified in Freund's incomplete adjuvant. Before and after the horses were inoculated, sera were collected sequentially and examined by radioimmunoassay. Rabbit anti-(horse) Ig and [125I]protein A served as the indicator system. Antigen-specific IgM, IgG, and IgA were observed to follow a classic immune response. The radioimmunoassay offers a valuable tool fo...
Gunther JD, Foley CW, Gaverick HA, Plotka ED.Progesterone concentrations were measured in milk and blood plasma for 15 mares throughout a normal estrous cycle and early pregnancy to determine the feasibility of utilizing progesterone in milk as an indicator of pregnancy. Samples were obtained daily from foaling until diagnosis of pregnancy by rectal examination at 30 to 35 days of gestation. Progesterone in milk and blood plasma was quantified by radioimmunoassay. Mean progesterone concentrations (+/- SE) in plasma from foaling to foal heat and during estrus, luteal phase and pregnancy were .51 +/- .09 ng/ml, .53 +/- .08 ng/ml, 3.88 +/- ...
Mathieu HP, Mathieu-Nast C, Vrignaud C.A rapid and low cost radioimmunologic procedure for progesterone assay in mare plasma is proposed. Radioimmunoassay is performed directly on 10 microliter of unextracted plasma. Free progesterone is adsorbed on dextran-charcoal, then the aqueous phase is decanted and extracted by 1 ml of scintillation fluid. Counting is performed directly on this two-phase system. Results are comparable to those obtained with radioimmunoassays using extracted plasma.
Solter PF, Farner S.The purpose of this study was to compare 2 different nonradioactive assay methods with a conventional radioimmunoassay (RIA) measuring the concentration of serum thyroxine (T4) in horses. Serum was obtained from 85 adult standardbred horses. The T4 concentration of each sample was analyzed by RIA, chemiluminescent enzyme immunoassay (CEI), and homogeneous enzyme immunoassay (HEI). The correlation between the HEI method and RIA method was significantly greater (r = 0.89) than the correlation between the CEI and the reference method (r = 0.53). In addition, the precision of the HEI method was si...
Seeger K, Thurow H, Haede W, Knapp E.A simple enzyme immunoassay (EIA) for the measurement of progesterone is described. Antibody against 11-OH-hemisuccinate-BSA is bound to polystyrene tubes. 11-OH-hemisuccinyl-beta-D-galactosidase is used as enzyme-coupled antigen and methylumbelliferyl-beta-D-galactoside as substrate. Concentrations down to 0.156 ng/ml plasm or amounts of 93 pg/tube are detectable. Probit analysis gave a linear relationship between log concentration and percentage of binding. A comparison of EIA and radioimmunoassay gave a correlation coefficient of 0.81. The assay is sufficiently sensitive to estimate progest...
Perkins NR, Threlfall WR, Ottobre JS.Existence of ultradian variation in serum progesterone concentration and the relation between progesterone and luteinizing hormone (LH) secretory patterns were investigated in nonpregnant and pregnant mares. Blood samples were taken every 15 minutes for a 24-hour period on day 8 of the estrous cycle and day 18 of pregnancy, respectively. Progesterone and LH concentrations were determined by radioimmunoassay. Progesterone was secreted in pulsatile manner in nonpregnant and pregnant mares. Luteinizing hormone also was secreted in a pulsatile manner in both groups of mares. There was little tempo...
Floris B, Bini PP, Nuvole P.The blood testosterone changes in three Giara stallions, average age 8 years old and average weight 183 Kgs, were checked weekly for a year using a radioimmunological method. The horses showed a highly seasonal hormonal activity concentrated in the first half of the year (winter and spring). In the second half of the year (summer and fall) the stallions showed low levels of testosterone. The highest level of the androgen hormone occurred in April, the lowest in October. The absolute levels of this hormone agreed substantially with the relative literature.
Sharma OP.The concentrations of PGF-2alpha in the peripheral blood of five foaling mares were measured by radioimmunoassay. Low levels of PGF-2alpha were detected as early as 1 week before foaling in two of the mares. These levels increased steadily, reaching a peak (1-74 +/- 0-44 ng/ml) during fetal expulsion. A relatively high PGF-2alpha level was found in samples collected 60 min after foaling.
Rossdale PD, McGladdery AJ, Ousey JC, Holdstock N, Grainger L, Houghton E.Maternal plasma progestagen concentrations increase about 20 days before parturition. The major contributors to the increase are reduced metabolites (ie 5 alpha-pregnanes). Precocious increases (ie less than 310 days of gestation) in these metabolites may occur in abnormal pregnancies. The effects of CRH, ACTH or betamethasone administered to the foetus at gestational ages ranging from about 250 to 320 days were examined. Sixteen healthy pony mares were used for foetal injection employing aseptic techniques. Water or normal saline were used as controls. Maternal plasma progestagen concentratio...
Bacigalupo MA, Ius A, Meroni G, Dovis M, Petruzzelli E.A time-resolved fluoroimmunoassay (TR-FIA) for the direct determination of clenbuterol residues in horse urine using a highly specific monoclonal antibody has been compared with an immunoenzymometric assay (IEMA). The sensitivity of both methods was 10 pg; the calibration curve was linear between 10 and 10(5) pg for the TR-FIA and between 10 and 10(4) pg for the IEMA.
Taylor PM, Luna S, Dye A.To determine the effects of surgery, hypoxia, hypercapnia and flunixin administration on plasma β-endorphin immunoreactivity (BEI) in anaesthetized horses. Methods: Prospective crossover study. Methods: Six healthy adult Welsh Mountain ponies and seven healthy adult Thoroughbreds. Methods: Ponies were anaesthetized with thiopentone and halothane or with pentobarbitone and the horses with guaiphenesin, thiopentone and halothane. Ponies were anaesthetized for 2 hours and on separate occasions underwent a period of hypoxia, hypercapnia, anaesthesia only, or were given flunixin at induction. The ...
Roser JF, Lofstedt RM.Blood and urine samples collected from 12 mares at frequent intervals from 25 to 210 d of pregnancy were analyzed for equine chorionic gonadotropin (eCG). Blood and urine samples were collected daily through two consecutive ovulatory periods from five cyclic mares for comparative purposes. Separate radioimmunoassays (RIA) were developed to detect eCG in the urine and plasma. A simple and quick commercial dipstick enzyme-linked immunospecific assay (ELISA), developed for eCG in the blood, was also utilized in this study to detect eCG in the urine. In the 12 pregnant mares, eCG concentrations in...
Rance TA, Park BK.Oestrone, equilin and dehydroepiandrosterone were measured by radioimmunoassay in peripheral plasma from five pony mares; samples were taken from day 60 of pregnancy at approximately weekly intervals through to parturition. Of the three steroids measured, dehydroepiandrosterone showed the most consistent pattern of secretion. DHA concentrations remained low until day 105, increased rapidly from day 120 and attained maximum values between days 168–210; levels declined to day 300, but there was considerable variation between mares in DHA patterns prior to parturition. There was great variation...
Bossut DF, Leshin LS, Malven PV.Radioimmunoassay procedures were developed and validated for the quantification of beta-endorphin (beta-EP)-like immunoreactivity in equine plasma. beta-EP could be quantitatively extracted from plasma with silicic acid powder and subsequently assayed, however, valid estimates of this hormone could also be obtained on unextracted plasma. Although beta-lipotropin (beta-LPH) cross-reacted in the assay, it was not necessary to correct for beta-LPH activity when assaying unextracted plasma because chromatographic analyses showed that 92% of the immunoreactivity in plasma extracts was similar in mo...
Benoit E, Garnier F, Courtot D, Delatour P.Antiserum has been raised in rabbits treated with a 19 nor testosterone-hemisuccinate-bovine-serum-albumin conjugate and used for the development of a specific RIA of plasma 19 nor testosterone. Plasma samples are drawn from testicular and jugular veins of stallions during castration under general anesthesia. Results demonstrate a testicular secretion of 19 nor testosterone and a stress inhibition of this secretion correlatively with stress inhibition of testosterone secretion.
Altmeyer P, Holzmann H, Stöhr L, Koch HJ.White horses are subject to age-dependent coat depigmentation. They are dark gray or black at birth and lose their coloring between their second and fourth year. Beginning at about age 10 their coat takes on a characteristic silver-gray coloring. The purpose of this paper was to find out to what extent the endogenic alpha-MSH level changes with the change in pigmentation. alpha-MSH plasma levels were determined by radioimmunologic analysis in 3 age groups of white Camarque horses: age group 1 consisted of dark horses with a mean age of 1.2 years and a mean alpha-MSH level of 106.4 pg/ml +/- 18...
Higgins AJ, Lees P, Taylor JB.In a two part cross-over experiment, acute inflammatory exudates were induced in 7 ponies by subcutaneous implantation of 3 sterile carrageenin-soaked polyester sponge strips. Treatment comprised a single therapeutic geenin-soaked polyester sponge strips. Treatment comprised a single therapeutic dose of 4.4 mg/kg phenylbutazone (PBZ) administered intravenously at the time of sponge implantation. Exudates were harvested at 6, 12 and 24 hours and examined for leukocyte and erythrocyte numbers using the improved Neubauer technique; for eicosanoids by radioimmunoassay and by high performance liqui...
