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.
Ware VC.In an attempt to understand more fully processes that control the selection or recruitment of follicles for ovulation, the superovulation paradigm in combination with the androgen, 5 alpha-dihydrotestosterone, or the antiandrogens, cyproterone or cyproterone acetate, was used in the immature mouse to alter the developmental potential of follicles destined to ovulate or to become atretic. Quantitative analysis of the numbers of eggs ovulated after one or more rounds of stimulation by pregnant mare's serum gonadotrophin followed by human chorionic gonadotrophin (PMSG-hCG), revealed a dose-depend...
Fletcher MA, Caldwell KE, Saez L, Latif Z.A sialoglycoprotein from horse erythrocytes was isolated in essentially homogeneous form and found to contain the neuraminidase-sensitive determinant of the horse erythrocyte for Paul-Bunnell heterophile antibodies of infectious mononucleosis. This reactivity was retained after covalent coupling of the antigen to latex particles. The latex reagent has greater stability (greater than 3 years) than either fresh or preserved horse erythrocytes. It can be used in a direct slide test; no absorption of the serum is necessary. The new test compared favorably with some standard tests for infectious mo...
Chapman DI, Close PM, Moss MS, Snow DH.A radioimmunological method was used as a screening procedure to determine the period of detection or "clearance time", for the horse, of therapeutic doses of the synthetic anabolic steroid nandrolone phenylpropionate. Seven horses, either at rest or being exercised, were given a course of weekly intramuscular injections of the steroid. On the separate occasion, some of the horses were given a single intramuscular injection of the same compound. The weekly injections maintained a high plasma concentration of nandrolone and/or metabolites. The mean (+/- sd) period of detection in plasma of thes...
Wilson MG, Nicholson WE, Holscher MA, Sherrell BJ, Mount CD, Orth DN.Using RIAs for six regions within proopiolipomelanocortin (proOLMC), gel filtration, and electrophoresis, we studied pituitary peptides in a normal horse and one with Cushing's disease caused by a pars intermedia adenoma. Almost all immunoreactive (IR) ACTH (78%) was 4,500 mol wt (4.5K) ACTH in normal pars distalis, but it was almost 100% corticotropin-like intermediate lobe peptide (CLIP) in normal pars intermedia. alpha MSH and beta MSH were found mainly in pars intermedia: equal concentrations of the beta MSH precursors, beta-lipotropin (beta LPH) and gamma LPH, were found in pars distalis....
Frauenfelder HC, Blevins WE, Page EH.Twelve periocular fibrous connective tissue sarcomas in 11 horses were treated with 222Rn. Follow-up periods ranged from 1 to 6 years; the overall nonrecurrence rate at 12 months after therapy was 92%. Two lesions recurred 2 years after treatment, and 1 after 3 years. One of the former lesions has not recurred after a 2nd 222Rn treatment.
Baker HW, Baker ID, Epstein VM, Hudson B.Cortisol and testosterone were measured by radioimmunoassay in plasma samples from race horses. None of 6 stressed male horses had low cortisol levels but testosterone levels (0.81 +/- 0.15 nmol/l) were significantly lower than in healthy horses (1.86 +/- 0.31 nmol/l). The conclusion was made that adrenocortical insufficiency is not a common association of stress in race horses. The reduction in testosterone levels is probably a nonspecific response similar to that seen in other species.
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...
Stewart DR, Stabenfeldt GH, Hughes JP.Plasma relaxin concentrations were measured hourly by radio immunoassay in 4 pregnant mares from 11 days before until 4 days after natural foaling. Pre-partum levels ranged from 4 to 7 ng/ml without any surge until the second stage of labour when they increased rapidly to about 11 ng/ml. In 3 of these mares, relaxin activity declined immediately after the expulsion of the placenta and was below detectable levels within 36 h. In the other mare relaxin activity did not fall until after the mechanical removal of the placenta 7 h after foaling. Eight mares were induced to foal by the administratio...
Alexander S, Irvine CH.Mares were bled once daily throughout a cycle, or 3 times daily from the first day of oestrus to the 2nd day after ovulation. LH was measured by heterologous radioimmunoassay and by an in-vitro bioassay based on LH-stimulated testosterone production by mouse Leydig cells. The patterns of bio- and immuno-active LH during the oestrous cycle were similar but not identical, so that in both groups of mares the ratio of biological: immunological (B:I) activity during the LH surge was significantly higher before than after ovulation (P less than 0 . 001). Considerable individual variation in cycle me...
Kindahl H, Knudsen O, Madej A, Edqvist LE.Blood samples from 4 mares during the late luteal phase, oestrus, early pregnancy and up to about 150 days of gestation were analysed for 15-keto-13,14-dihydroprostaglandin F-2 alpha (PGFM), progesterone, PMSG and oestrone sulphate by radioimmunoassays. During the late luteal phase, at the time of corpus luteum regression and decreasing progesterone levels, PGFM peaks were recorded. During early pregnancy (i.e. from mating and up to about Day 30) no such peaks were detected. After mating the progesterone levels increased and remained high throughout the observation period. During the oestrous ...
Irvine CH, Alexander S.Serum LH was measured by radioimmunoassay in 5 long-term (greater than or equal to 3 years) castrated male horses bled at 30-min intervals for 2 h twice a month from March to February. There was no significant effect of month on LH levels; however, mean levels were significantly lower in spring than autumn (P less than 0.05). By contrast, stallions in the same environment showed a markedly seasonal pattern of LH secretion, with LH rising at the onset of the breeding season to reach levels in late spring 3-4 times those in early winter. Despite differences in seasonal patterns of secretion, ann...
Evans JW, Roser JF, Mikuckis GM.Human chorionic gonadotrophin (hCG) can be used to study horse luteinizing hormone (LH) receptors in stallion testicular tissue. hCG was more stable than horse LH during radioiodination when compared by their abilities to bind to testicular receptor sites. During incubation, neither hormone lost binding activity at 4 degrees C. Horse LH lost binding activity during incubation at 25 degrees C and both hormones lost binding activity at 37 degrees C. Both hormones bound to the same receptor sites which are specific for the hormones. The receptor sites were not degraded when incubated at 4 degrees...
Williams DL, Issel CJ, Steelman CD, Adams WV, Benton CV.Biological and mechanical transmission trials with Psorophora columbiae (Dyar and Knab) and Aedes sollicitans (Walker) and ponies acutely infected with equine infectious anemia virus (EIAV) were negative. The EIAV antigen was detected by radioimmunoassay in Ae sollicitans immediately after the mosquitoes had fed on an acutely ill pony, but not 14 days after feeding. Psorophora columbiae mosquitoes had detectable EIAV antigen as determined by radioimmunoassay 24 hours after they fed on an acutely ill pony; this antigen was not detected again until 6 days after feeding and was still detected 14 ...
Chen CL, Riley AM.Serum thyroxine (T-4) and triiodothyronine (T-3) concentrations were assayed in neonatal foals (1.5 to 4 months) and mature horses (2 to 25 years old) by a modified radioimmunoassay procedure. Blood was collected from 52 clinically healthy foals and horses of various breeds (Thoroughbreds, Quarter Horses, American Saddle Horses, and a single cross-bred horse). Neonatal foals had high serum concentrations of T-4 (mean, 4.02 microgram/dl) and T-3 (192.9 ng/dl) as compared with the values in mature horses (T-4, mean of 1.76 microgram/dl; T-3, mean of 98.69 ng/dl). Stallions had slightly higher T-...
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 +/- ...
Fillion G, Beaudoin D, Rousselle JC, Jacob J.Glial cell membrane fractions were prepared using glial cells preparations isolated from horse brain striatum. [3H]5-HT binding was measured by the filtration technique and the adenylate cyclase activity determined by measuring the cAMP production using a radioimmunoassay. Serotonin binds to glial membrane fractions with an affinity corresponding to a dissociation constant Kd = nM. The corresponding site is serotoninergic specific: [3H]5-HT binding is inhibited by 5-HT agonists (5 OH NM-DMT, 5-MeOHT, 5-MeOH-DMT, NN-DMT) or antagonists (cinanserine, cyproheptadine, methysergide, LSD) and not (o...
Chapman DI, Moss MS, Tomlinson PW, Harrison MP, Simmons PJ.1. Following single intramuscular doses of [14C]fluprostenol (0.5--2.4 micrograms/kg) to three female horses and to three gelded male horses, radioactivity was present in the plasma within 5 min; peak concn. (0.32--1.30 ng/ml fluprostenol equiv.) occurred 5 to 90 min after injection. Radioactivity was still present in the plasma of the females after three days. About 88% of fluprostenol is bound to plasma proteins. 2. Radioactivity was present in the parotid saliva of the gelded male horses within 10 min. Peak concn. (45--91 pg/ml fluprostenol equiv.) occurred from 5 min to 1 h after injection...
Button C, Gross DR, Johnston JT, Yakatan GJ.The pharmacokinetics of IV administered digoxin and the bioavailability of intragastrically administered powdered digoxin tables suspended in water were investigated in 6 clinically normal adult horses by 125I radioimmunoassay. The effect of 3 to 5 sequential IV doses of 5 micrograms of digoxin/kg of body weight at 2-hour intervals on a left ventricular index of contractility (Vmax) was assessed in 5 clinically normal horses. Standard pharmacokinetic equations and mean pharmacokinetic variables were used to derive parenteral and oral (loading and maintenance) doses for digoxin in horses. The c...
Aggarwal BB, Farmer SW, Papkoff H, Stewart F, Allen WR.Serum of the pregnant donkey, like that of the mare, contains a gonadotrophin of chorionic origin. The chorionic gonaditrophin of the donkey (dCG) has been isolated in purified form from the serum of pregnant donkeys using methodology previously employed for the purification of pregnant mare chorionic gonadotrophin (eCG). Unlike eCG, dCG is predominatly an LH in biological tests. In the in-vitro rat Leydig cell assay, dCG was as active as eCG, but in the in-vitro rat seminiferous tubule assay for FSH and in the augmentation assay, dCG was considerably less potent than eCG (1-10%). Specific rat...
Pedersoli WM, Belmonte A, Purohit RC, Ravis WR.The pharmacokinetics of gentamicin were studied in six healthy mature horses of mixed breeding and of both sexes. A parenteral preparation of gentamicin sulfate (5% aqueous solution) was administered rapidly (IV) at the dosage level of 5 mg/kg of body weight. Venous blood samples were taken at 0 (base line), 0.083, 0.25, 0.5, 0.75, 1, 2, 4, 8, 12, 24, 36, 48, 72, 96, and 120 hours after gentamicin administration. Serum gentamicin was measured by a radioimmunoassay technique. The gentamicin concentration data was fitted to a one- and two-compartment open model with first-order elimination from ...
Purohit RC, Nachreiner RF, Humburg JM, Norwood GL, Beckett SD.Circulating concentrations of angiotensin I and plasma renin activity (PRA) were determined in 45 Thoroughbreds before prerace medication (at rest), 1 to 1 1/2 hours before racing but after prerace medication (prerace), within 15 minutes after racing (immediately postrace), and between 1 to 2 hours after racing. Angiotensin I was measured by radioimmunoassay, and PRA was determined by the rate of generation of angiotensin I in plasma incubated at 37 C. Irrespective of prerace medication, there was a marked increase in angiotensin I (0.478 +/- 0.034 to 0.848 +/- 0.051 ng/ml) and PRA (0.191 +/- ...
Moore JN, Steiss J, Nicholson WE, Orth DN.In the horse, a syndrome of hirsutism, hyperglycemia, glucosuria, polydipsia, polyuria, polyphagia, and progressive debilitation has been recognized. Most often the syndrome has been associated with adenomas of the pars intermedia of the pituitary and bilateral adrenal hyperplasia or nodular hyperplasia involving primarily the zona fasciculata. Previously, the syndrome has been ascribed to compression of the hypothalamus by an expanding but functionally inactive pituitary neoplasm. In the present case, with RIA determination of plasma ACTH concentrations, the syndrome was ascribed to pituitary...
Menzer C, Schams D.A double-antibody radioimmunoassay for PMSG, especially for meauring PMSG in cattle blood after exogenous application, has been developed. A rabbit antiserum against PMSG and pure PMSG for radioiodination were used. There was a strong cross-reaction against equine LH and FSH, but the slight cross-reaction against bovine LH and FSH could be eliminated by adding bovine LH to each tube during the assay. Unspecific, interfering influences of equine or cow serum could be eliminated by adding a constant amount of PMSG-free serum to each tube. PMSG added to 200 microliter of serum could be recovered ...
Charman H, Long C, Coggins L.Three structural proteins of equine infectious anemia virus were purified, labeled with 125I, and utilized in radioimmunoassays with horse sera and antisera to heterologous retroviruses. Whereas radioimmunoassay titers for the major protein, p25, were 500- to 1,000-fold higher than titers in immunodiffusion, for clinical purposes these two procedures were equivalent. Antibodies to two low-molecular-weight proteins, p12 and p10, were also found in infected horses, but with a lower frequency and lower titers. As a rule, only sera positive for p25 also contained antibody to p12 and p10. Antisera ...
Zavy JT, Mayer R, Vernon MW, Bazer FW, Sharp DC.Uterine flushings were collected from 30 non-pregnant Pony mares on Days 8, 12, 14, 16, 18 or 20 after oculation. Mares were allowed a recovery period of one oestrous cycle and were mated at the next oestrus. They were then ovario-hysterectomized on days which corresponded to the day of the oestrous cycle to which they were assigned. Uterine flushings were analysed for total recoverable protein and acid phosphatase activity. Least squares analysis indicated a status X day interaction for total protein (P less than 0.10) and acid phosphatase activity (P less than 0.005) in which the latter was ...
Raeside JI, Liptrap RM, McDonell WN, Milne FJ.Plasma levels of total oestrogens and dehydroepiandrosterone (DHA) were measured by radioimmunossay in samples taken from various blood vessels in both maternal and fetal compartments in 11 Pony mates. High concentrations of oestrogens (greater than 100 ng/ml of plasma), expressed as oestrone equivalents, were found in the fetal circulation. On both the fetal and maternal sides, oestrogen concentrations were lower in blood going to than from the placenta. DHA concentrations, on the other hand, were higher in blood flowing to the placenta from the fetus. The fetal gonads were seen as the source...
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...
Terqui M, Palmer E.Plasma total (conjugated + unconjugated) oestrogens were measured from Day 0 to 100 of pregnancy and compared with the levels found during the oestrous cycle. From Day 0 to 35 of gestation, the concentrations were similar to those during dioestrus. An increase in total oestrogens between Days 35 and 40 was followed by a plateau of 3 ng/ml between Days 40 and 60 which was slightly higher than preovulatory concentrations. This first increase in total oestrogen level was produced by the ovaries since values were suppressed after ovariectomy; stimulation may be due indirectly to PMSG causing folli...
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...
Aggarwal BB, Farmer SW, Papkoff H, Stewart F, Allen WR.Serum of the pregnant donkey, like that of the mare, contains a gonadotrophin of chorionic origin. The chorionic gonaditrophin of the donkey (dCG) has been isolated in purified form from the serum of pregnant donkeys using methodology previously employed for the purification of pregnant mare chorionic gonadotrophin (eCG). Unlike eCG, dCG is predominatly an LH in biological tests. In the in-vitro rat Leydig cell assay, dCG was as active as eCG, but in the in-vitro rat seminiferous tubule assay for FSH and in the augmentation assay, dCG was considerably less potent than eCG (1-10%). Specific rat...
Tai CL, Wang C, Weckman TJ, Popot MA, Woods WE, Yang JM, Blake J, Tai HH, Tobin T.To improve the sensitivity and specificity of screening for etorphine in horses, an 125I-labeled etorphine analog was synthesized and an antibody to etorphine was raised in rabbits. A radioimmunoassay (RIA) for etorphine was developed, using these reagents. Bound and free 125I-labeled etorphine was separated by a double-antibody method that reduced interference from materials associated with equine urine. The 125I-labeled etorphine binding was rarely greater than 250 pg of background etorphine equivalents/ml in raw urine and was 100 pg/ml in hydrolyzed urine. The 125I-RIA was capable of detect...
Woods WE, Weckman T, Wood T, Chang SL, Blake JW, Tobin T.A commercially available radioimmunoassay kit was used to screen for the presence of etorphine in post-race urines from horses racing in Kentucky. Most horse urines contained small amounts of materials which reacted positively in this immunoassay. These materials are apparently endogenous to the horse and were called apparent etorphine equivalents. The levels of these apparent etorphine equivalents in post-race urines from 70 horses were estimated. Their modal level averaged 0.1 ng/ml, the population distribution was log normal, and individual horses showed levels of up to 0.8 ng/ml.
Rasbech NO, Koefoed-Johnsen HH, Holm G.On the basis of progesterone determination on plasma or blood after RIA and the kit method respectively and consecutive scanning performed on a total number of 31 mares the following features were demonstrated: The overall material shows that in 20 mares (64.5%) embryonic vesicles were demonstrated. Of these mares 16 have conceived after service in the 1st estrous cycle and 3 mares in the 2nd estrous cycle. 18 mares were scanned in the time interval 13th-26th day after the latest service, while 2 mares were scanned on day 46 and 41 respectively. A total of 14 scanning positive mares were exami...
Carstanjen B, Sulon J, Banga-Mboko H, Beckers JF, Remy B.This study describes for the first time the development and validation of a sensitive and specific radioimmunoassay (RIA) for equine osteocalcin (OC) quantification using purified equine OC as standard, tracer, and immunogen for antibody formation in rabbits. The assay allowed to measure equine serum OC levels with a sensitivity of 0.2 ng/mL. Immunoreactive serum OC values of clinically normal, different-aged horses ranged from 3.68 to 127.31 ng/mL. Intra- and inter-assay coefficients of variation (CV) were 6.2 and 8.2%, respectively. Serial equine serum sample dilutions were linear. The recov...
Tangyuenyong S, Nambo Y, Nagaoka K, Tanaka T, Watanabe G.Most thyroid hormone determinations in animals are based on immunoassays adapted from those used to test human samples, which may not reflect the actual values of thyroid hormone in horses because of the presence of binding proteins. The aims of the present study were i) to establish a novel radioimmunoassay (RIA) using a more simple and convenient method to separate binding proteins for the measurement of total thyroxine (T4) in horses and ii) to validate the assay by comparing total T4 concentrations in yearling horses raised in different climates. Blood samples were collected from trained y...
Cesar FB, Stewart AJ, Boothe DM, Ravis WR, Duran SH, Wooldridge AA.Nine horses received 20 mg/kg of intravenous (LEV ); 30 mg/kg of intragastric, crushed immediate release (LEV ); and 30 mg/kg of intragastric, crushed extended release (LEV ) levetiracetam, in a three-way randomized crossover design. Crushed tablets were dissolved in water and administered by nasogastric tube. Serum samples were collected over 48 hr, and levetiracetam concentrations were determined by immunoassay. Mean ± SD peak concentrations for LEV and LEV were 50.72 ± 10.60 and 53.58 ± 15.94 μg/ml, respectively. The y-intercept for IV administration was 64.54 ± 24.99 μg...
Young DW, Smyth GB.The predominant form of gastrin in the antral mucosa of the stomach of virtually all species previously examined is the 17 amino acid peptide little gastrin (G17). This report describes the occurrence in equine antral mucosa of an immunoreactive form of gastrin with elution properties on Sephadex G-50 superfine similar to human unsulfated big gastrin (G34-I). This putative equine big gastrin was a major component of the gastrin immunoreactivity present. A second peak of activity in equine antral mucosa eluted in an identical manner to human little gastrin (hG17-I). Inhibition curves of equine ...
Raeside JI, Christie HL.C(18) neutral steroid formation by cytochrome P450 aromatase has been recorded for several equine and porcine tissues. High activity of P450 aromatase is reflected in the quantities of estrogens in yolk-sac (y-s) fluid of early equine conceptuses. In a previous study of y-s fluid we detected large amounts of androgens by radioimmunoassay (RIA), using an antiserum for androstenedione (A(4)). Here, we report that RIA, following chromatography, gave tentative identification of the major peak as norandrostenedione (19-norA) not as A(4). Furthermore, even greater quantities of 19-norA seemed to be ...
Thompson DL, McNeill DR, Wiest JJ, St George RL, Jones LS, Garza F.Sequential samples of blood were drawn via jugular catheters every 15 min for 24 h from four mares in each of five reproductive states: intact anestrous mares in winter, intact diestrous mares in summer, intact estrous mares in summer, ovariectomized mares in winter and ovariectomized mares in summer. Estrous mares were sampled on d 4 or 5 of estrus and diestrous mares on d 10 or 11 of diestrus. Each sample of plasma was assessed for concentrations of luteinizing hormone (LH) and follicle stimulating hormone (FSH) in two independent radioimmunoassays. A computer program was developed that dete...
McKeever KH, Hinchcliff KW, Cooley JL, Lamb DR.The purpose of this study was to test the hypothesis that furosemide administration before exercise would cause greater increases in plasma arginine vasopressin (AVP) concentration in exercising horses than exercise alone. Six adult, clinically normal, unfit mares underwent three randomly ordered 60 minute standard exercise tests on an equine treadmill to examine the effect of furosemide administration on plasma AVP concentration. In one trial, furosemide (1 mg kg-1) was infused four hours before exercise (FUR-4) and a placebo (10 ml saline) was infused two minutes before exercise; in another ...
Morris DD, Crowe N, Moore JN.The purpose of this study was to determine if a structurally novel dual inhibitor of arachidonic acid metabolism, SK & F 86002, would inhibit the endotoxin-induced production of tumor necrosis factor (TNF) activity by equine peritoneal macrophages. Equine peritoneal macrophages were variously pretreated for 0, 0.5 and 2 h with SK & F 86002 at 10(-9) to 10(-4) molar final concentrations or were left untreated. Then, the macrophages were cultured in vitro in the presence of endotoxin (5 ng/mL). Supernatant media were collected after 4 h and stored at -70 degrees C until assayed for TNF a...
Carstanjen B, Amory H, Youssao I, Remy B.The purpose of this study was to validate an equine-specific osteocalcin (OC) radioimmunoassay (RIA) for use in donkeys and to establish age-related changes in serum OC concentrations in healthy donkeys. Serial dilutions of donkey serum showed parallelism with standard curves obtained with the equine-specific OC RIA. There was a tight linear regression between donkey serum OC values obtained with the equine specific OC RIA and a commercially available bovine-specific OC RIA. Serum OC levels of 27 healthy donkeys, analysed with the equine-specific OC RIA, showed a tight negative logarithmic reg...
Alexander S, Irvine CH.Mares were bled once daily throughout a cycle, or 3 times daily from the first day of oestrus to the 2nd day after ovulation. LH was measured by heterologous radioimmunoassay and by an in-vitro bioassay based on LH-stimulated testosterone production by mouse Leydig cells. The patterns of bio- and immuno-active LH during the oestrous cycle were similar but not identical, so that in both groups of mares the ratio of biological: immunological (B:I) activity during the LH surge was significantly higher before than after ovulation (P less than 0 . 001). Considerable individual variation in cycle me...
Wood T, Tai CL, Taylor DG, Woods WE, Wang CJ, Houtz PK, Tai HH, Weckman TJ, Yang JM, Sturma L.Detomidine is a potent non-narcotic sedative agent which is currently in the process of being approved for veterinary clinical use in the United States. Since no effective screening method in horses is available for detomidine, we have developed an 125I radioimmunoassay for detomidine in equine blood and urine as part of a panel of tests for illegal drugs in performance horses. Our 125I radioimmunoassay has an I-50 for detomidine of approximately 2 ng/ml. Our assay shows limited cross-reactivity with the pharmacodynamically similar xylazine, but does not cross-react with acepromazine, epinephr...
Thompson DL, Johnson L.Prolactin concentrations in stallion serum were measured by radioimmunoassay based on antiserum generated against equine prolactin and radioiodinated canine prolactin. Prolactin concentrations in serum collected from 152 stallions at a slaughterhouse were higher (P less than .01) in summer than in winter (5.7 +/- .15 vs 2.0 +/- .17 ng/ml). Moreover, there was an effect of age (P less than .02) in the analysis of variance; there was no interaction between age and season. In general, prolactin concentrations increased with age up to 3 to 5 years. Samples of serum collected from five control and ...
Hoque S, Senba H, Tsunoda N, Derar RI, Watanabe G, Taya K, Osawa T, Miyake Y.To clarify the endocrinological characteristics of the mares with granulosa theca cell tumor (GTCT), peripheral plasma samples from the 6 mares affected with GTCT were collected before and after the surgical removal of the affected ovary. Concentrations of testosterone (T), follicle stimulating hormone (FSH), luteinizing hormone (LH), immunoreactive-inhibin (ir-INH), progesterone (P) and estradiol-17beta (E(2)) in the plasma samples were measured by radioimmunoassay. Before removal of GTCT in all cases, the concentrations of T were significantly higher than those of normal mares at the breedin...
Roussel AJ, Lin YC, Strait JR, Modransky PD.Radioimmunoassay for parathyroid hormone (PTH) in equids was performed on blood samples from healthy equids and equids with hypercalcemia and hypophosphatemia. The assay was validated for equine carboxy-terminal PTH. Manipulation of serum ionized Ca in healthy equids by infusing Na2 EDTA and CaCl2 produced an expected increase and decrease, respectively, in measurable immunoreactive PTH. Intra-assay and interassay coefficients of variation were 2.6% and 11.7%, respectively. The range of PTH valves for healthy mature horse mares and geldings maintained on pasture was less than 0.27 ng/ml to 0.9...
Sigler DH, Ericson DE, Gibbs PG, Kiracofe GH, Stevenson JS.Lactating mares were assigned as controls or fed altrenogest (.044 mg.kg body wt-1.d-1) for 15 d after foaling. Mares (n = 6) fed altrenogest were inseminated during the first estrus after treatment and mares (n = 6) in the control group were inseminated during the second postpartum estrus. Ovulation during the estrus in which mares were inseminated occurred 26 +/- 1 d postpartum for treated mares and 36 +/- 1 d postpartum for control mares. The percentage of mares conceiving was not different for control (67%) and alternogest-treated (100%) mares. No differences were observed in tone and size...
Karsai F, Szaniszló F, Pethes G.The levels of the two primary bile acids, cholic acid (CA) and chenodeoxycholic acid (CDCA), were determined by radioimmunoassay in cattle, horse, pig and dog serum. The mean serum cholic acid (SCA) and deoxycholic acid (SCDCA) levels of cows varied with their reproductive status, being 7.8 (+/- 3.3) and 1.5 (+/- 1.0) mumol/l in dry cows, 17.8 (+/- 6.9) and 2.3 (+/- 1.0) mumol/l in freshly calved dams, and 15.8 (+/- 5.7) and 2.3 (+/- 0.8) mumol/l, respectively, in lactating cows. The SCA level found in the immediate prepartal period and also on the day of calving corresponded to those found du...
Doff SC, Wenderlein J, Wiesinger A, Hiereth S, Ulrich S, Straubinger RK.Lyme borreliosis is a vector-borne disease in humans and animals caused by bacteria from the sensu lato complex (sl). The possible transmission of sl from companion animals to humans via ticks makes this disease important in terms of One Health approaches. Thus, early and accurate diagnosis and treatment are of utmost importance. Today's standard for the detection of specific antibodies against sl is a two-tiered test system based on an ELISA for screening combined with a line immunoassay (LIA) for confirmation. In this study, 200 canine and 200 equine serum samples with known antibody status...
Geschwind II, Dewey R, Hughes JP, Evans JW, Stabenfeldt GH.Plasma levels of LH were determined by heterologous radioimmunoassay utilizing highly purified equine LH as standard. Samples were taken regularly from eleven mares for twenty-six oestrous cycles over a period of 10 weeks. The mean cycle length was 20-5 +/- 3-1 (S.D.) days, and ovulation occurred on average 4-3 +/- 1-6 (S.D.) days from the time heat was first detected. Levels of LH were persistently low from Days 5 to 16 of the cycle (ovulation = Day 0). They then increased slowly over a number of days and continued to rise beyond the levels observed at any time during the immediate preovulato...
Tucker KE, Henderson KA, Duby RT.Twenty-three follicles were collected from 14 mares on specific days and grouped to represent follicles from early (Group 1; n = 6), mid (Group 2; n = 11) and late (Group 3; n = 6) oestrus, as described previously (Tucker et al., 1988). Isolated granulosa cells (GC) from each follicle were cultured in multiwell plates containing either Eagle's Minimum Essential Medium (MEM) alone, eLH (300 ng/ml), eFSH (300 ng/ml) or eLH + eFSH (300 ng/ml each), in the presence or absence of 0.5 microM testosterone. Media were collected and replaced at 24 h of culture, and 24 h later, media were again collecte...
Motton DD, Roser JF.Recent evidence in our laboratory suggests that the cause of idiopathic subfertility/infertility in breeding stallions may originate in the testes at the luteinizing hormone (LH) receptor or postreceptor level. The objective of this research was to determine if LH receptor binding activity is altered in subfertile and infertile stallions. Six fertile, three subfertile, and three infertile stallions, ages 11-23 years, were classified according to normal semen parameters and pregnancy rates and then castrated in the breeding season. Blood was collected prior to castration, and plasma was stored ...
Champion ZJ, James EA, Vickers MH, Breier BH, Casey PJ.The effect of intramuscularly administered recombinant bovine growth hormone (rbGH) on insulin-like growth factor-I (IGF-I) and white and red blood cell indices was studied in Thoroughbred geldings. An insulin-like growth factor binding protein (IGFBP)-blocked radioimmunoassay was modified and validated for the measurement of IGF-I in equine blood plasma. Baseline values of IGF-I and blood indices were determined over a 48 h period and then a single dose of 5 microg/kg, 10 microg/kg or 50 microg/kg of rbGH was administered. Insulin-like growth factor-I levels increased in a dose-dependent mann...
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 ...
Sayegh AI, Anderson NV, Harding JW, Cerpovicz P, DeBowes RM, Ritter RC, Baker GJ, Reeck G.To purify and characterize pepsinogens in equine gastric mucosa. Methods: Stomachs collected from 2 healthy horses at necropsy. Methods: After collection, stomachs were placed immediately in ice before storage at -48 C. After slow thawing, the mucosa was scraped off while the tissue was immersed in 0.1M potassium phosphate (pH 7.4) at 4 C, then was homogenized. The filtered extract was subjected to anion-exchange chromatography. Fractions that were found to contain pepsin or pepsinogen were further chromatographed. Individual fractions were tested for pepsinogen or pepsin content by monitoring...
Lanci A, Mariella J, Ellero N, Canisso IF, Dondi F, Castagnetti C.This study aimed to determine alpha-fetoprotein (AFP) concentrations in amniotic fluid, plasma of mares and respective foals: carrying normal pregnancies and delivering healthy foals (n = 20; Group 1); carrying apparently normal pregnancies and delivering sick foals (n = 15; Group 2); carrying high-risk pregnancies and delivering sick foals (n = 14; Group 3). High-risk pregnancy was defined by a history of premature udder development/lactation or increased of the combined thickness of the uterus and placenta, or vulvar discharge and/or mares' systemic illness. Sick foals were affected by...
Ohya T, Kondo T, Yoshikawa Y, Watanabe K, Orino K.In mammal circulation, various ferritin-binding proteins (FBPs) are thought to be involved in the clearance of circulating ferritin after complex formation with it. However, horse FBPs are known to cause inhibitory effects on ferritin immunoassay due to the concealment of the ferritin molecule to anti-ferritin antibodies used in the ferritin immunoassay. These inhibitory effects are eliminated by heat treatment of horse serum at 75°C for 15 min. The inhibitory effects on ferritin immunoassay in the sera of ten foal sera (5 females and 5 males) from 1 to 18 months were detected during all peri...
