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.
Weber JA, Woods GL, Freeman DA, Vanderwall DK.Prostaglandin E2 (PGE2) secreted by Day-6, Day-7, Day-8 and Day-9 equine embryos (ovulation = Day 0) during in vitro incubation was measured by radioimmunoassay. Embryonic PGE2 secretion (ng/embryo/24 hr) was detectable on Day 6 (0.27 +/- 0.39), tended to increase (P less than 0.1) on Day 7 (0.57 +/- 0.88), and increased significantly (P less than 0.05) on Day 8 (2.23 +/- 0.86) and Day 9 (4.13 +/- 0.71). Embryo diameter at the start of the incubation period was linearly correlated (P less than 0.01) to embryonic PGE2 secretion.
Stevenson KR, Parkinson TJ, Wathes DC.Jugular venous blood samples were collected throughout a complete oestrous cycle from 9 mares for measurement of progesterone and oxytocin by radioimmunoassay. Mean oxytocin concentrations remained at approximately 1 pg/ml throughout, with no evidence of cyclic variation in the release pattern. Extracts of corpus luteum and follicles obtained from a further 33 mares at different stages of the cycle all contained oxytocin concentrations of less than 10 pg/g wet weight of tissue. We conclude that the ovaries are not a source of circulating oxytocin during the oestrous cycle in this species. The ...
Whitehair KJ, Parker JE, Smith GN, Adams SB, Bottoms GB.Serum levels of type III procollagen peptide (P-III-P) were measured by radioimmunoassay in clinically normal adult ponies (n = 15) and horses (n = 10). The mean serum levels of P-III-P from the ponies, 10.4 +/- 2.9 (SD) ng/mL, and the horses, 12.2 +/- 2.6 (SD) ng/mL, were not significantly different. Segments of jejunum were made ischemic to induce fibrous peritoneal adhesions in two ponies, and serum P-III-P levels were measured on days 4, 5, 7, 14, and 21. An exploratory celiotomy on day 21 revealed that the ischemic injury had induced fibrosis of the mesentery and bowel, but no adhesions h...
McFarlane JR, Coulson SA, Papkoff H.This study describes the presence of immunoactive and bioactive eCG-like material in full-term placentas of both domestic horses and zebras. Term placental extracts were immunoreactive in an LH monoclonal antibody RIA, and methods successfully used previously for the purification of eCG and eLH were employed to further concentrate the immunoreactive materials to the point where additional characterization studies could be performed. Sufficient equine material was obtained to perform a final fractionation on a concanavalin A Sepharose column yielding an unadsorbed fraction, e17A, and an adsorbe...
Reimers TJ, Lamb SV, Bartlett SA, Matamoros RA, Cowan RG, Engle JS.Veterinary diagnostic endocrinology laboratories frequently receive hemolyzed plasma, serum, or blood samples for hormone analyses. However, except for the previously reported harm done by hemolysis to canine insulin, effects of hemolysis on quantification of other clinically important hormones are unknown. Therefore, these studies were designed to evaluate effects of hemolysis on radioimmunoassay of thyroxine, 3,5,3'-triiodothyronine, progesterone, testosterone, estradiol, cortisol, and insulin in equine, bovine, and canine plasma. In the first experiment, hormones were measured in plasma obt...
McFarlane JR, Czekala NM, Papkoff H.Six samples of pregnant zebra (z) serum from the first and second trimesters of pregnancy were analyzed by RIA and shown to have chorionic gonadotropin levels comparable to that of the mare (0.9-5.3 micrograms/ml); first trimester levels in most cases were higher than second trimester levels. A pool of the sera (10 ml) was fractionated by methods previously employed for the purification of equine (e) and donkey (d) chorionic gonadotropin to achieve a concentration of the zebra chorionic gonadotropin (zCG). A yield of 1.0 mg of glycoprotein was obtained. HPLC analysis of the material indicated ...
Stewart F, Tuffnell PP.A 514 bp cDNA transcript coding for 78% of horse (Equus caballus.) GH has been cloned and sequenced. The deduced amino acid sequence corresponded precisely to that previously obtained by protein sequencing and, in addition, provided new sequence information for the signal peptide. The missing 3' fragment of the cDNA was reconstructed using synthetic oligonucleotides and site-specific directed mutagenesis. The complete cDNA sequence was then inserted into an expression vector (PIN-III-lppp-5) which utilizes a bacterial signal peptide to secrete the expressed product into the periplasmic space o...
Teale P, Houghton E.A screening procedure for anabolic steroid residues in horse urine has been developed based upon solid-phase extraction and gas chromatographic/mass spectrometric analysis in the selected ion mode. For moderate sample throughput the method provides a viable alternative to radioimmunoassay screening and has advantages over the latter technique due to its flexibility, specificity and ability to detect a number of steroids in a single analysis. Full automation of the gas chromatographic/mass spectrometric analysis is an additional feature of the methodology.
Hines KK, Affleck KJ, Barrows SP, Murdoch WL, Fitzgerald BP, Loy RG.The relationship between daily mean FSH concentrations in serum and the pattern of FSH detected by frequent sampling for 12-h periods (samples every 15 min) was examined in five mares during the transition into the breeding season. The five mature anestrous mares were exposed to a natural increase in daylength. Blood samples were collected daily from February 1 until the first ovulation of the breeding season (April 14 +/- 3.7 days, Mean +/- SEM). Periods of frequent blood collection were performed every two weeks. Blood samples were obtained daily by jugular venipuncture or jugular cannula (f...
Nostrandt AC, Pedersoli WM, Marshall AE, Ravis WR, Robertson BT.Ototoxicosis was evaluated in 6 healthy ponies given 5 mg of gentamicin/kg of body weight, q 8 h, IM. Ponies 1, 2, and 3 were dosed for 7 days and ponies 4, 5, and 6 were dosed for 14 days. Serum peak and trough concentrations of gentamicin were measured by radioimmunoassay at regular intervals. Brain stem auditory-evoked responses were recorded every 5 days up to 60 days after the first dose to monitor auditory function. Although serum gentamicin concentrations were within or above the accepted clinical therapeutic range, loss of auditory function was not observed at the frequency range (1 to...
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...
Johnson L, Varner DD, Thompson DL.To determine the age at which adult values for hormonal and testicular characteristics are established, testes and blood from 123 slaughtered horses, aged 1-5 years, were compared. Serum concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and testosterone (T) and intratesticular testosterone (ITT) were determined by radio-immunoassay. Specimens were obtained in the non-breeding (n = 46) or breeding (n = 43) season in Experiment 1 (Exp. 1) and in the breeding (n = 34) season in Experiment 2 (Exp. 2). Testicular parenchyma was fixed in glutaraldehyde and homogenized to...
Santschi EM, LeBlanc MM, Weston PG.Plasma cortisol, oestrone sulphate and progestagens were measured in 22 stressed pregnant mares (gestation length 17-336 days) as indicators of fetal viability. Mares were bled every 12 h from time of admission, and plasma was stored at -70 degrees C until assayed. Four normal mares were bled twice weekly from Day 270 to parturition to provide baseline endocrine data. Cortisol and progestagen concentrations were measured by radioimmunoassay and oestrone sulphate was measured by enzyme immunoassay. Mares were grouped according to clinical diagnosis: surgical colic (Group 1, n = 11), medical col...
Baldwin DM, Roser JF, Muyan M, Lasley B, Dybdal N.Enzymatically dispersed anterior pituitary cells from donor mares were cultured for 48 h in alpha-modified Eagles' medium containing 10% steroid-free horse serum. The cells were then incubated for 24 h in fresh medium oestrogen followed by a 4-h incubation with or without GnRH. Media and cell extracts were analyzed for LH by radioimmunoassay. In the first series of experiments, pituitary cells from Day-3 dioestrous mares were preincubated with ethanol (control) or different concentrations of E2 (10(-11) to 10(-7) M) for 24 h prior to a 4-h incubation without (basal) or with 1.0 nM GnRH. E2 inc...
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...
Silver M, Fowden AL, Knox J, Ousey J, Cash R, Rossdale PD.Pre- and post partum changes in plasma T3 have been examined in relation to plasma cortisol in 23 newborn foals (12 full term, 6 premature and 5 'twilight'), and in 5 fetuses catheterized in late gestation. Blood samples were collected daily from the fetuses and from the neonates at 30-min intervals for 2 h after birth; the plasma was assayed for T3 and cortisol by standard radioimmunoassay methods. In the full-term foals, plasma cortisol and T3 concentrations were high at birth (67.4 +/- 6.1 and 4.3 +/- 0.3 ng/ml respectively) and rose to a maximum during the following 2-h period to 141.0 +/-...
Houghton E, Holtan D, Grainger L, Voller BE, Rossdale PD, Ousey JC.Radioimmunoassay (RIA) and gas chromatography-mass spectrometry (GC-MS) were used to determine plasma progestagen concentrations in the normal and premature foal. Radioimmunoassay provides a profile of plasma progestagens with respect to time but, due to the non-specific nature of the technique and without prior chromatographic purification, quantitative data based on RIA analysis must be interpreted with caution. In contrast, the greater specificity of GC-MS allows identification of specific plasma progestagens and measuring of multiple analytes in a single analysis. Both techniques demonstra...
Pantke P, Hyland J, Galloway DB, MacLean AA, Hoppen HO.Equine plasma luteinizing hormone (LH) possesses both biological (in vitro bioassay, B) and immunological (radioimmunoassay, I) activities and the ratio of B:I varies with stage of the oestrous cycle. To estimate the contribution made by pituitary secretion and peripheral metabolism to changes in the B:I ratio, pituitary venous effluent and circulating plasma from 5 dioestrous and 2 oestrous mares were analyzed using both an in vitro bioassay and a radioimmunoassay. During dioestrus, LH was released in a pulsatile fashion with a frequency of 1.4 pulses/24 h and a pulse duration of 20-40 min (c...
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...
Perryman LE, O'Rourke KI, Mason PH, McGuire TC.Equine-murine xenohybridoma cells were produced using SP2/0 murine myeloma cells and splenic lymph node cells obtained from horses infected with 10(6) TCID50 of single cloned variants of equine infectious anaemia virus (EIAV). The xenohybridomas secreted equine IgG monoclonal antibodies reactive with EIAV in enzyme immunoassays employing purified virus. Seven antibodies were studied in detail. They bound to viral glycoproteins (gp90 or gp45) in radioimmunoprecipitation assays, and reacted with homologous EIAV as well as five other cloned variants of EIAV. When evaluated against a single cloned...
Hyland JH, Langsford DA.Foetal death was induced in 10 Standardbred mares at day 45 of gestation by injecting 20 to 45 ml of hypertonic (24% W/V) saline into the conceptus at surgery. Ten mares underwent sham treatment and acted as controls. Blood and urine samples were collected every other day between days 30 and 45 post ovulation and at 0, 3 and 6 h relative to the infusion of saline in the treated mares, or sham treatment in control mares. Blood and urine samples were then collected daily between days 46 and 55 post ovulation. Urine oestrone sulphate (E1S) concentrations, measured by radioimmunoassay, increased b...
Meinecke B, Gips H.The aim of the present investigations was to characterize the endocrine changes in the peripheral plasma during the periparturient phase of mares with a known history of obstetrical disorders. Blood plasma samples from 9 mares (8 mares during parturition, 1 mare during abortion) were collected and the following steroid hormones were radioimmunologically determined: progesterone (P4), dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEA-S), estrone (E1), estrone sulfate (E1-S), estradiol-17 beta (E2), estriol (E3), cortisol and transcortin. In general, with the exception of corti...
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...
Nequin LG, King SS, Matt KS, Jurak RC.The transition from anoestrus to oestrus in mares is controlled by photoperiod. The present study examined whether additional daylength would accelerate the mares' response to gonadotrophin-releasing-hormone (GnRH). Nine anoestrous mares were placed under ambient or artificial long lighting on 7th January. The four month experimental period was divided into a three-day sequence which was repeated at 21 day intervals. Ovaries were palpated rectally on Day 1; saline was injected (1 ml intravenously [iv]) on Day 2; GnRH was administered (0.59 microgram/kg bodyweight iv) on Day 3. Blood was taken ...
Adolf GR, Traxler E, Maurer-Fogy I.Equine interferon-beta 1 (EqIFN-beta 1) was purified from extracts of recombinant Escherichia coli by sequential chromatography on hydroxylapatite, anion-, and cation-exchangers. The resulting protein was greater than 98% pure as determined by sodium dodecylsulfate gel electrophoresis, gel permeation HPLC, and reverse-phase HPLC. Amino-terminal amino acid sequencing revealed that essentially all molecules contained an additional amino-terminal methionine. The specific antiviral activity of EqIFN-beta 1 determined on equine dermal fibroblasts challenged with vesicular stomatitis virus (VSV) was...
Tulamo RM, Saari H, Konttinen YT.Concentration of hyaluronate (HA) in equine serum was determined by a recently developed specific radioassay. The mean +/- SD HA concentration in equine serum was 288 +/- 145 micrograms/L, was age dependent, and varied widely between horses (range, 190 to 760 micrograms/L). Light or moderate exercise increased serum HA concentration from baseline values by 1.5- to 3-fold. In all horses, serum HA concentration returned to or below the original resting values 1 and 2 hours after exercise.
Lepage OM, Marcoux M, Tremblay A.Levels of alkaline phosphatase and osteocalcin or bone Gla-protein, a new marker of bone metabolism, were analyzed in blood samples of 50 clinically normal female Standardbred horses between four months and twenty years of age. Samples were collected in the morning before exercise. Serum osteocalcin was measured by radioimmunoassay using bovine antibodies. There was a significant inverse correlation between alkaline phosphatase, osteocalcin and the age of the animals up to 48 months. The decrease in osteocalcin levels in serum was very marked during the first 30 months of life. The mean osteoc...
Adam A, Ong H, Sondag D, Rapaille A, Marleau S, Bellemare M, Raymond P, Giroux D, Loo JK, Beaulieu N.A monoclonal antibody was synthesized in mouse against the O-(3-carboxypropionyl) derivative of albuterol linked to bovine serum albumin. Isotyping of this material revealed the IgG1 class characterized by an affinity constant of 1.03 nM-1 and a density of sites of 0.55 nM. This antibody was found specific as its cross-reactivity to structurally related molecules was less than 1% except for clenbuterol (75%). A radioimmunoassay was set up with culture supernatant (final dilution 1/1000) and [3H] albuterol. The calibration curve was characterized by a maximum binding of 28%, an ED50 of 1.15 pmo...
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 ...
Melrose PA, Walker RF, Douglas RH.Catecholamine concentrations (pg/ml) in the cerebrospinal fluid (CSF) of prepubertal (n = 9) and adult (n = 18) horses were determined by radioenzymatic assay. Norepinephrine was low or non detectable in all CSF samples. In contrast, measurable CSF dopamine concentrations were effected by age, reproductive status and exogenous steroid treatments. The concentration of dopamine in the CSF of prepubertal females (733 +/- 92) was greater (p less than 0.05) than the concentration in the CSF of prepubertal males (117 +/- 67). Prepubertal male horses which were treated with testosterone for 5 days (5...
May SA, Hooke RE, Lees P.Although an E-series prostaglandin has previously been identified in equine inflammatory exudate, the identity of this eicosanoid as PGE2 has not been confirmed. The objective of this study was the specific characterisation of the prostaglandin produced by equine cells in the presence of an inflammatory stimulus. By using two radioimmunoassays, a relatively non-specific PGE2 assay and a more specific PGE1 assay, it has been possible to identify the E-series prostaglandin produced by equine chondrocytes and synovial cells, in response to a variety of stimuli, as PGE2.
Nett TM, Holtan DW, Estergreen VL.The levels of LH, prolactin and oestrogen were measured by radioimmunoassay in serum obtained from six mares during the post-partum period, and LH was also assayed in blood samples collected at 6-hr intervals from four normal cyclic mares during oestrus and at daily intervals during dioestrus. The concentrations of oestrogen and LH suggested that the mares were having ovulatory cycles during the post-partum period irrespective of behavioural oestrus, and the levels of prolactin seemed to rise in association with probable ovulatory LH peaks. In these mares levels of LH had increased significant...
Dembek K, Timko K, Moore C, Johnson L, Frazer M, Barr B, Toribio R.Sepsis is a major cause of morbidity and mortality in neonatal foals. Relative adrenal insufficiency (RAI), defined as an inadequate cortisol response to stress, has been associated with sepsis, prematurity, and poor outcome in newborn foals. In addition to cortisol, the adrenal gland synthesizes several biologically important steroids and steroid precursors, including aldosterone, androgens, and progestogens. However, concentration of these hormones during hospitalization and their association with the severity of disease and mortality in critically ill foals have not been completely evaluate...
Hoffman LS, Adams TE, Evans JW.Blood samples were obtained from 5 mixed-breed, long-term castrated geldings during five 24-h periods between May 1984 and April 1985. Blood samples were collected, beginning at 09:00 h, at 15-min intervals for 8 h and hourly for the remaining 16 h. Plasma concentrations of LH and FSH were determined by RIA. Seasonal changes in hormone concentrations and frequency and amplitude of secretory pulses were evaluated. No diurnal variation in either LH or FSH secretion was observed: however, marked circhoral fluctuations in LH and FSH secretion were noted. Mean LH and FSH concentrations in these lon...
Stewart DR.Equine relaxin (eRlx) immunoactivity has previously been measured in the mare during pregnancy using the porcine relaxin (pRlx) RIA (pRlx-RIA). This was not the optimal system for measurement of eRlx because the dose-response curve obtained with equine plasma was not parallel to the pRlx standard curve. A homologous eRlx-RIA has been developed and used to measure relaxin immunoactivity during pregnancy and parturition in the mare. Highly purified eRlx was used for the generation of antiserum in rabbits, preparation of tracer, and as assay standards. A double antibody eRlx RIA (eRlx-RIA) was de...
Schumacher SA, Toribio RE, Lakritz J, Bertone AL.The objective of this study was to characterize the pharmacokinetics and pharmacodynamics of intravenous administration of magnesium sulfate to horses using a novel radio-telemetry system for physiologic signal capture. Five Horses were surgically implanted with a radio-telemetric carotid catheter. Implants were paired with a non-invasive telemetric unit which acquired a six lead ECG and 3-axis acceleration to assess activity acquired wirelessly in real-time for future analysis. Horses were exposed to a new stall environment before (baseline) and after 60 mg/kg (30 mL) of magnesium sulfate (Mg...
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...
Mendoza FJ, Buzon-Cuevas A, Toribio RE, Perez-Ecija A.Insulin dysregulation (ID) is diagnosed in horses and ponies using oral glucose (OGTT) and oral sugar (OSTT) tolerance tests. The enteroinsular axis plays a major role in postprandial glucose disposal and insulin response in horses, ponies and foals. The insulin and incretin response to oral carbohydrate challenges has not been characterised in donkeys. Objective: (a) To characterise OGTT and OSTT, and (b) to assess the plasma incretin response to OGTT and OSTT in healthy donkeys. Methods: In vivo experiments. Methods: Six healthy adult female Andalusian donkeys were challenged with OGTT (1 g...
Satué K, Domingo R, Redondo JI.The coordinated interaction of the components of the renin angiotensin aldosterone system (RAAS) with reproductive hormones such as progesterone, oestrogens and cortisol during pregnancy has been widely reported to play a vital role in foetal and placental development in various species, significantly influencing the proper achievement of pregnancy and foetal viability at birth. These interactions have not yet been clarified in mares. Thus, the purpose of the present research was to analyse the relationship between cortisol (CORT), progesterone (P4) and oestrone sulphate (OESTRONE), and the co...
Bonnaire Y, Plou P, Pages N, Boudene C, Jouany JM.A highly sensitive procedure for GC/MS determination of etorphine in horse urine is described. This assay provides both specificity and reliability and is particularly well suited for the confirmation of radioimmunoassay screening procedures usually used for etorphine. After solvent extraction and purifications, the etorphine is characterized as a pentafluoroacetic derivative (PFAA) by using mass fragmentography. The detection limit is 0.1 ng/mL in urine; the coefficient of variation of the estimations is 10.9%. The procedure has been validated after on-field administration of 5 to 90 microgra...
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...
Roser JF, Tarleton M, Belanger JM.Recent studies in our laboratory investigating plasma gonadotrophin concentrations before and after castration indicate that the hypothalamic-pituitary axis is not the original site of dysfunction in stallions with idiopathic infertility. In the present study, fertile, subfertile and infertile stallions were subjected to replacement therapy with oestradiol and testosterone 1 year after castration to investigate hypothalamic-pituitary axis function further. Fertile (n=5), subfertile (n=3) and infertile (n=2) castrated stallions of Light horse breeds, aged 11-23 years, were given either oestradi...
Aurich C, Sieme H, Hoppe H, Schlote S.To investigate the involvement of endogenous opioids in the regulation of gonadotrophin release in male horses, effects of the opioid antagonist naloxone (0.5 mg kg-1 i.v.) on plasma LH and testosterone concentrations and the possible influence of season and of gonadal steroids were investigated. To determine quantitative as well as qualitative changes in gonadotrophin release, LH concentrations were measured by radioimmunoassay and by an in vitro bioassay. Experiments were performed in May, August and December. In stallions, basal LH secretion in May and August was significantly higher than i...
Aurich C, Schlote S, Hoppen HO, Klug E, Hoppe H, Aurich JE.To investigate an involvement of endogenous opioids in the regulation of circannual changes in reproductive activity, effects of the opioid antagonist naloxone on the concentration of immunoreactive and bioactive luteinizing hormone (LH) in plasma were measured in mares during the anovulatory season. Naloxone (0.5 mg/kg i.v.) caused a significant increase (P < 0.05) in immunoreactive as well as bioactive LH concentration in plasma. The amplitude of the increase in LH concentrations measured with an in vitro bioassay was more pronounced than the amplitude of the increase in LH secretion dete...
Leme DP, Papa FO, Roser JF.The objective of this study was to investigate reproductive characteristics of stallions at a tropical zone in the breeding and non-breeding seasons. The following parameters were assessed: testicular volume; semen quality; and serum concentrations of LH, FSH, and testosterone; in addition to the percentages of germ cells and proportions of germ cells/Sertoli cells by testicular cytology in stallions. Semen was collected from eight adult stallions twice a week during a 12-week period in both seasons (6 weeks before and 6 weeks after the summer and winter solstices). Jugular blood samples were ...
Nourian AR, Mills PC, Pollitt CC.Microdialysis (MD) was used for continuous monitoring of the lamellar extracellular fluid (ECF) in six mature healthy Standardbred horses. MD probes were introduced into the lamellar tissue under local anaesthesia. Following intravenous injection of gentamicin (5mg/kg), MD and serum samples were collected for 24h and analysed using a sensitive ELISA test for gentamicin and fluorescence polarization immunoassay for urea concentrations. Calibration of probes was performed through in vivo urea recovery and in vitro gentamicin and urea recovery. Data obtained from different body compartments were ...
Stewart F, Goode JA, Allen WR.A heterologous radioimmunoassay was developed and validated for the measurement of horse GH in plasma. It utilized recombinant-derived bovine GH as the radiolabelled ligand, a guinea-pig anti-porcine GH serum as first antibody and pituitary-derived horse GH as standard. Cross-reactivities were high with all of the pituitary and recombinant-derived GH preparations tested (49-140%) and very low (< 0.3%) with horse FSH, LH and prolactin. A synthetic analogue of GH-releasing factor(1-29) stimulated the expected pattern of GH release in foals. Plasma GH concentrations in foals were low at birth (< ...
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...
Cambridge H, Reynoldson JA, Dunsmore JD.Plasma thromboxane B2 (TXB2) the stable inactive metabolite of thromboxane A2 (TXA2), was measured daily by specific radioimmunoassay in three groups of animals before and after experimental infection with Strongylus vulgaris. Infection of four 'parasite naive' foals produced a typical acute syndrome with intermittent but statistically insignificant rises in TXB2 levels. Interpretation of results was complicated by the presence of a non-septic peritonitis associated with implantation of the foals with electrodes for recording myoelectrical activity. In two foals of similar age, with some natur...
McFarlane JR, Czekala NM, Papkoff H.Six samples of pregnant zebra (z) serum from the first and second trimesters of pregnancy were analyzed by RIA and shown to have chorionic gonadotropin levels comparable to that of the mare (0.9-5.3 micrograms/ml); first trimester levels in most cases were higher than second trimester levels. A pool of the sera (10 ml) was fractionated by methods previously employed for the purification of equine (e) and donkey (d) chorionic gonadotropin to achieve a concentration of the zebra chorionic gonadotropin (zCG). A yield of 1.0 mg of glycoprotein was obtained. HPLC analysis of the material indicated ...
Coyne CP, Hornof WJ, Kelly AB, O'Brien TR, DeNardo SJ.Two methods were analyzed for the rapid extraction of equine fibrinogen from fresh plasma, using ammonium sulfate-sodium phosphate buffer. Fibrinogen from each of these 2 methods was then radiolabeled with 125I (half-life = 60.2 days, gamma = 35 keV), using monochloroiodine reagent. Mean protein-bound activity was 98.5% and mean clottable radioactivity was 94.1%. Radiolabeled fibrinogen administered IV to 15 horses had an overall mean (+/- SD) plasma half-life of 4.95 +/- 0.44 days.
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 ...
Czerwinski SL, Lyon AW, Skorobohach B, Léguillette R.Ophthalmic antibiotic therapy in large animals is often used empirically because of the lack of pharmacokinetics studies. The purpose of the study was to determine the pharmacokinetics of topical tobramycin 0.3% ophthalmic solution in the tears of normal horses using an automated immunoassay analysis. Results: The mean tobramycin concentrations in the tears at 5, 10, 15, 30 minutes and 1, 2, 4, 6 hours after administration were 759 (±414), 489 (±237), 346 (±227), 147 (±264), 27.6 (±28.4), 14.8 (±66.6), 6.7 (±18.6), and 23.4 (±73.4) mg/L. Mean tobramycin concentration was maintained abo...
Chen CL, Goldberg J, Gronwall RR.A radioimmunoassay was developed for prednisolone using IgG purified from rabbit antiserum. The assay was employed to determine the pharmacokinetics of prednisolone following intravenous administration of 450 mg of prednisolone sodium succinate (Solu Delta Cortef) to five adult Thoroughbred horses. The RIA had a sensitivity of 2 ng/ml and was relatively specific. It had cross-reactivity with 21-deoxycortisol (83.3%) cortisol (27.8%), 11-beta-hydroxyprogesterone (39.2%) and 17-hydroxyprogesterone (50%). However, it did not cross-react with naturally occurring steroids (cholesterol, testosterone...
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 ...
Silberzahn P, Pouret EJ, Zwain I.Androgen (testosterone and androstenedione) and oestrogen (oestradiol -17 beta and oestrone) concentrations were measured by radio-immunoassay in the peripheral plasma of two geldings (five-years-old), three bilateral cryptorchids (two, two and a half, and five-years-old) and three normal intact stallions (four, five and five and a half-years-old) before and after a single injection of 10,000 iu human chorionic gonadotrophin (hCG). In the stallions, hCG administration resulted in an immediate sharp increase of conjugated oestrogens and a more gradual increase of unconjugated androgens. In the ...
Raz T, Gray A, Hunter B, Card C.Superovulatory treatment may potentially increase the embryo recovery rate and the per-cycle pregnancy rate in normal or subfertile mares that are managed properly. However, some studies suggest a possible negative effect of superovulatory treatment on ovarian follicular maturation and embryo viability. Objectives of the present study were to investigate the early effects of eFSH treatment in reproductively normal mares in terms of: folliculogenesis, pregnancy rate, early embryonic development, reproductive tract parameters (tone and edema), and serum estradiol-17beta and progesterone concentr...
Rigoleto Júnior WL, Dias de Camargo Neto W, de Paula Nogueira G, Ferrari TA, Bertan Membrive CM, Giometti IC, Castilho C.The objective of this study was to quantify serum progesterone levels, uterine features, and pregnancy rates in acyclic, embryo recipient mares using a bovine progesterone-releasing intravaginal device in a commercial embryo transfer (ET) program. The study included 73 recipient mares of unknown breed, aged 3-10 years, weighing 350-500 kg, and kept under an intensive management system on Tifton 85 (Cynodon spp.) pastures with water and mineral salt ad libitum. The horses were divided into two groups: a group with a progesterone-releasing intravaginal device (1 g progesterone, G-IVP4, n = 24...
Trachsel DS, Schwarzwald CC, Grenacher B, Weishaupt MA.Measurement of atrial/A-type natriuretic peptide (ANP) concentrations may be of use for assessment of cardiac disease, and reliable data on the analytic performance of available assays are needed. To assess the suitability for clinical use of commercially available ANP assays, intra-assay and inter-assay coefficient of variation and dilution parallelism were calculated for three immunoassays (RIAPen, RIAPhoen, and an ELISAPen) using blood samples from healthy and diseased horses to cover a wide range of ANP concentrations. Further, agreement between assays was assessed using linear regression ...
Teale P, Houghton E.A screening procedure for anabolic steroid residues in horse urine has been developed based upon solid-phase extraction and gas chromatographic/mass spectrometric analysis in the selected ion mode. For moderate sample throughput the method provides a viable alternative to radioimmunoassay screening and has advantages over the latter technique due to its flexibility, specificity and ability to detect a number of steroids in a single analysis. Full automation of the gas chromatographic/mass spectrometric analysis is an additional feature of the methodology.
