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.
Lemazurier E, Moslemi S, Sourdaine P, Desjardins I, Plainfosse B, Seralini GE.The steroid content of semen from a total of 11 mature fertile stallions was studied during two breeding seasons and one winter. The levels of free and conjugated substrates (testosterone and androstenedione), and products (estradiol and estrone), of aromatase were measured by radioimmunoassay with a validated method. The results were seasonally and monthly highly variable with characteristic peaks. The concentrations of free and conjugated estrogens were always higher in the gel-free ejaculate than in the gel except in one subfertile stallion used as comparison. Furthermore, the steroid produ...
Champion ZJ, Breier BH, Ewen WE, Tobin TT, Casey PJ.A survey of standardbred horses was conducted to build up a normal population profile for insulin like growth factor-I (IGF-I) concentrations in racing standardbreds and to ascertain how age, sex and geographic location affect IGF-I. Blood samples were drawn by jugular venepuncture from 202 racing standardbred horses aged one to eight years located in five different geographic regions of New Zealand. IGF-I concentrations were determined by insulin like growth factor-I binding protein (IGFBP)-blocked radioimmunoassay validated for the horse. As described in other species, age played a significa...
Duggan JM, Coates DM, Ulaeto DO.Venezuelan equine encephalomyelitis (VEE) virus is an important human and veterinary pathogen of Central and South America. The virus can cause widespread epidemics, affecting hundreds of thousands of horses, and thousands of humans. Detection of the virus early in infection and in mosquito populations may allow epidemics to be predicted such that suitable prophylaxis, such as vaccination, can be used to reduce disease severity and transmission. The sensitivity and specificity of current immunoassays, based on conventional monoclonal and polyclonal antibodies, needs to be improved for the diag...
Donadeu FX, Ginther OJ.The role of the number of follicles and circulating immunoreactive inhibin in the decrease in plasma FSH concentrations that occurs during development of a follicular wave was studied in mares. All follicles > or = 6 mm in diameter were ablated by ultrasound-guided transvaginal aspiration of follicular fluid on day 10 after ovulation. During the subsequent wave, all follicles, the three largest follicles (three follicle group), the largest follicle (single follicle group) or no follicles were retained and the remaining follicles were ablated before they reached > 10 mm in diameter (n = 1...
Robion FC, Doizé B, Bouré L, Marcoux M, Ionescu M, Reiner A, Poole AR, Laverty S.In vivo the effects of intra-articular (IA) corticosteroids on articular cartilage remain controversial. This study was designed to examine this issue using synovial fluid (SF) markers of cartilage metabolism. Paired radiocarpal joints, without clinical or radiographic signs of joint disease, were studied in 10 adult horses. Aseptic arthrocentesis was performed weekly for 13 weeks. IA injections of methylprednisolone acetate (MPA) into the treatment joint and the vehicle into the control joint were performed at weeks 3, 5 and 7. We used radioimmunoassays on SF samples which measure a keratan s...
Henderson K, Stewart J.A dipstick, competitive immunoassay for rapidly measuring serum oestrone sulfate (OS) concentrations in horses was developed to distinguish mares 100 or more days pregnant from non-pregnant animals. 6-Ketoestrone 6-carboxymethyloxime conjugated to bovine serum albumin (oestrone CMO-BSA) was 'dotted' 25 mm from the bottom edge of 45 x 5 mm strips of polyester-film-supported cellulose nitrate membrane, pore size 3 microm. The strips were blocked, dried and a 15 x 5-mm cellulose absorbent sink attached 10 mm from the top of each strip. The manufactured dipsticks were stored with desiccant at room...
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...
Kirker-Head CA, Chandna VK, Agarwal RK, Morris EA, Tidwell A, O'Callaghan MW, Rand W, Kumar MS.To correlate substance P content of synovial fluid with prostaglandin E2 content, radiographic evidence of joint abnormality, and anatomic location of the joint for normal and osteoarthritic joints of horses. Methods: Synovial fluid from 46 normal joints in 21 horses and 16 osteoarthritic joints in 10 horses. Methods: Normal and osteoarthritic joints were identified by clinical and radiographic examination, by response to nerve blocks, during scintigraphy or surgery, or by clinicopathologic evaluation. Substance P and prostaglandin E2 contents of synovial fluid were determined by radioimmunoas...
Kearns CF, Lenhart JA, McKeever KH.Erythropoietin (EPO) is the primary hormone of erythropoiesis. Administration of recombinant human erythropoietin (rhuEPO) to improve racing performance in the horse represents a new form of blood doping, which has been associated with increased mortality. While immunoassay kits have become plentiful, very few commercial hormone assays are made specifically for equine research. There is a strong degree of sequence homology reported for EPO among species, which has allowed antibodies designed for human EPO research to be used to determine EPO concentration in other species. The objective of the...
Chiba S, Kanematsu S, Murakami K, Satoh A, Asahina M, Numakunai S, Goryo M, Ohshima K, Okada K.Serum parathyroid hormone (PTH) and calcitonin (CT) levels in fractured racehorses were measured by radioimmunoassay. Racehorses with fracture of large bone such as the radius, third metacarpus, third carpus, digital bone or tibia, showed normal PTH level and elevated CT level in the serum. Serum PTH level was slightly higher in racehorses with sesamoid bone fracture compared to that of healthy racehorses, but not statistically significant. Moreover, serum CT level of racehorses with sesamoid bone fracture was significantly higher than that of healthy racehorses. Racehorses with sesamoid bone ...
Frean SP, Lees P.To investigate effects of the anti-arthritic agents hyaluronan and polysulfated glycosaminoglycan (PSGAG) on inflammatory metabolism in cultured equine synoviocytes. Methods: Synoviocytes cultured from samples obtained from the metacarpophalangeal joints of 4 horses. Methods: Equine synoviocytes were grown in monolayer culture. Synoviocytes were stimulated with lipopolysaccharide (LPS) and simultaneously treated with various concentrations of hyaluronan or PSGAG for 48 hours. Three hyaluronan preparations were compared. Prostaglandin E2 (PGE2) concentrations in culture medium were measured, us...
Lohmann KL, Roussel AJ, Cohen ND, Boothe DM, Rakestraw PC, Walker MA.To evaluate the correlation between halftime of liquid-phase gastric emptying (T50), determined with nuclear scintigraphy using technetium Tc 99m pentetate, and absorption variables of orally administered acetaminophen. Methods: 6 mature horses. Methods: Technetium Tc 99m pentetate (10 mCi) and acetaminophen (20 mg/kg of body weight) were administered simultaneously in 200 ml of water. Serial left and right lateral images of the stomach region were obtained with a gamma camera, and T50 determined separately for counts obtained from the left side, the right side and the geometric mean. Power ex...
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...
McKeever KH, Malinowski K.Six young (mean + s.e., 5.3 +/- 0.8 years, 445 +/- 13 kg bwt) and 6 old (22.0 +/- 0.4 years, 473 +/- 18 kg bwt) Standardbred and Thoroughbred mares were used to test the hypothesis that age would alter the endocrine response to exercise. All of the mares were unconditioned but accustomed to the laboratory, to standing quietly and running on a treadmill, and to the standardised incremental exercise test (SET) used in the experiment. Two weeks prior to the experiment, each horse underwent a SET to determine maximal oxygen uptake (VO2max) and the speeds to be used in the actual experiment. A seco...
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...
Faber NF, Roser JF.Testicular biopsy was performed as a standing procedure on fertile, mature stallions (n=7) under sedation and local anaesthesia. The 'Biopty' instrument was used to collect three samples from the left testis of each stallion. The oestradiol, testosterone and inhibin content in each testis were assessed by validated radioimmunoassay (RIA); protein concentrations were determined and the testes were prepared histologically for haematoxylin and eosin staining. The fertility status of each stallion was assessed before and after testicular biopsy by semen evaluation, changes in total scrotal width, ...
Watson ED, Buckingham J, Björkstén T, Nikolakopoulos E.The presence of oxytocin in the mare uterus was determined by radioimmunoassay of tissue extracts and uterine lavage fluid, and by immunohistochemistry. Samples were obtained from mares during the oestrous cycle and on day 14 of pregnancy. Immunoreactive oxytocin was detected in extracts of endometrium and myometrium, and in uterine lavage fluid. In tissue sections, the luminal epithelium, the epithelium of the superficial, but not deep, glands, blood vessel walls and myometrium all stained positively for oxytocin, whereas only the luminal epithelium and epithelium of the superficial glands st...
Behrendt-Adam CY, Adams MH, Simpson KS, McDowell KJ.In this study, the roles of oestrogen and progesterone in the regulation of oxytocin gene expression in equine endometrium were examined. Anoestrous mares (n=19) were assigned randomly to one of the following treatment groups: control (vehicle control for 1 day; n=3); progesterone (250 mg progesterone per day for 6 days; n=4); oestradiol (5 mg beta-oestradiol 17-valerate per day for 6 days; n=4); oestradiol plus short duration progesterone (5 mg beta-oestradiol 17-valerate per day for 6 days followed by 250 mg progesterone per day for 6 days; n=4); and oestradiol plus long duration progesteron...
Lebelt D, Zanella AJ, Unshelm J.The aim of the present study was to obtain information on the possible mechanisms underlying cribbing behaviour in horses. To investigate the horse's responsiveness to an external stimulus, a device for telemetric measurement of thermal threshold, using the forelimb withdrawal reflex, was developed and validated. Measurements of thermal threshold took place in cribbing horses (n = 11) before and during cribbing periods. Heart rate was monitored continuously in the same horses. Blood samples were collected before and during cribbing periods as well and in age- and sex-matched control horses (n ...
Yamaguchi K, Sawada T, Naraki T, Igata-Yi R, Shiraki H, Horii Y, Ishii T, Ikeda K, Asou N, Okabe H, Mochizuki M, Takahashi K, Yamada S, Kubo K....The prevalence of Borna disease virus (BDV)-specific antibodies among patients with psychiatric disorders and healthy individuals has varied in several reports using several different serological assay methods. A reliable and specific method for anti-BDV antibodies needs to be developed to clarify the pathological significance of BDV infections in humans. We developed a new electrochemiluminescence immunoassay (ECLIA) for the antibody to BDV that uses two recombinant proteins of BDV, p40 and p24 (full length). Using this ECLIA, we examined 3,476 serum samples from humans with various diseases ...
Chiappe A, Gonzalez G, Fradinger E, Iorio G, Ferretti JL, Zanchetta J.In this study, we assessed the potential value of free serum osteocalcin or bone gla protein (BGP), the most abundant non collagenous matrix protein found in bone and dentin, to reflect changes of bone turnover in thoroughbred horses. Levels of osteocalcin were analyzed in serum samples of 54 clinically normal animals divided into three groups (A, B, C) according to age: 8, 16-18 and 24-36 months, in order to determine the standard for young horses of different age and sex. Serum BGP was measured by an in-house developed double antibody radioimmunoassay using bovine antigen. The mean BGP level...
Paccamonti DL, Pycock JF, Taverne MA, Bevers M, Van Der Weijden GC, Gutjahr S, Schams D, Blouin D.We investigated the half-life of oxytocin in reproductively normal mares and the prostaglandin response after oxytocin administrations. Mares were given oxytocin, 10 or 25 iu, i.v., on the day of, or 2 days after, ovulation, and frequent jugular blood samples were collected for analysis of oxytocin and Prostaglandin F metabolite (PGFM) by RIA. Neither dose of oxytocin nor day of treatment affected the half-life of the exogenous oxytocin, which was determined to be 6.8 min. A significant increase in PGFM was observed within 6 min of oxytocin administration and peak values were observed within 1...
Spiess BM, Nyikos S, Stummer E, Sahin A, Naegeli H.To determine concentrations of dexamethasone in serum and urine of horses treated repeatedly with a topically administered ophthalmic dexamethasone preparation. Methods: 4 clinically normal horses (2 mares, 2 geldings). Methods: 0.1% dexamethasone ophthalmic ointment was administered to the left eye of each horse every 5 to 9 hours for 8 consecutive days, yielding an estimated cumulative dexamethasone dose of 6.4 microg/kg of body weight. Serum and urine samples were obtained before the first dexamethasone treatment, on days 4 and 8 of treatment, and 24, 48, and 96 hours after cessation of tre...
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.
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...
Armstrong S, Lees P.To examine effects of carprofen (enantiomers and a racemic mixture) on the metabolism of equine chondrocytes. Methods: Cartilage from clinically normal horses. Methods: Effects of carprofen on proteoglycan neosynthesis, glycosaminoglycan (GAG) release and prostaglandin (PG) E2 production by unstimulated chondrocyte monolayers and cartilage explants were examined, as were similar variables in monolayers and explants exposed to carprofen and recombinant human interleukin 1beta (IL-1). Carprofen (enantiomers and racemic mixture) was used alone or along with IL-1 on monolayers and explant cultures...
Deby-Dupont G, Grülke S, Caudron I, Mathy-Hartert M, Benbarek H, Deby C, Lamy M, Serteyn D.The strangulated intestinal pathologies of horses are accompanied by a local activation of the neutrophils, that can be revealed by measuring the tissular enzymatic activity of the granulocytic enzyme myeloperoxidase (MPO). To estimate the possible spreading of this neutrophil activation to the systemic circulation, we designed a radioimmunoassay (RIA) for equine neutrophil myeloperoxidase (MPO) (EC 1.11.1.7) using a specific rabbit antiserum. MPO was labeled with 1 mCi 125I by a technique of self-labeling in the presence of 10(-4) M hydrogen peroxide. The RIA was performed by incubation of 10...
McFarlane D, Sellon DC, Gaffney D, Hedgpeth V, Papich M, Gibbs S.To compare hematologic and serum biochemical variables and plasma ACTH concentration between healthy horses 5 to 12 years old and those more than 20 years old. Methods: 30 healthy horses 5 to 12 years old and 30 healthy horses more than 20 years old. Methods: Venous blood was collected from all horses, and CBC and serum biochemical analysis were performed for each horse. Plasma ACTH concentration was determined by radioimmunoassay. Student's paired t-test or the Mann-Whitney rank sum test was used to compare values between control and aged horse groups. Results: Compared with values for contro...
Nagy P, Solti L, Kulcsár M, Reiczigel J, Huszenicza G, Abaváry K, Wölfling A.Several assay systems (3H radioimmunoassay (RIA) with and without extraction; microplate enzyme-linked immunoassay (ELISA); qualitative ELISA (tube test)] were used to measure plasma progesterone concentration in mare plasma. The direct RIA showed a close correlation (R = 0.94) with the extraction RIA. The direct RIA and the microplate ELISA were compared in two different studies. In the first study 1155 samples of postpartum mares were used for progesterone determination with both assays. The ELISA resulted in more elevated values both in oestrus and dioestrus (0.19+/-0.3 and 2.44+/-3.62 nmol...
Kamerling S, Wood T, DeQuick D, Weckman TJ, Tai C, Blake JW, Tobin T.Narcotic analgesics produce pharmacological effects by interacting with specific opiate receptors. At least five major types of opiate receptors have been recognised. These include mu (morphine) and kappa (ethylketazocine) receptor types. Narcotic analgesics which interact with mu receptors produce locomotor and autonomic stimulation at doses that produce little or no analgesia. Therefore, use of these drugs as analgesics in equine medicine has not been very satisfactory. Theoretical considerations suggested that the role of kappa agonists in equine analgesia be investigated. Using a pure kapp...
Allen WE, Porter DJ.Milk and plasma samples were obtained every 48 hours from eight pony mares for 40 days after foaling. Progestogen concentrations in milk and plasma were measured using an enzyme-linked immunoassay (ELISA) and compared with radioimmunoassay of the plasma. In general the three assays showed similar trends in progestogen concentration changes but absolute values varied considerably. Difficulty could occur in interpreting the results from single samples taken at times when progestogen concentrations were either rising (ie, after ovulation) or falling. ELISA could be used on plasma obtained by allo...
Moore RM, Charalambous AC, Masty J.To measure colonic arterial (CA) and colonic venous (CV) plasma neuropeptide concentrations during low-flow ischemia and reperfusion of the large colon in horses. Methods: 10 adult horses. Methods: CA and CV plasma samples collected from anesthetized horses during experimentally induced low-flow colonic ischemia and reperfusion were assayed for vasoactive intestinal polypeptide (VIP), calcitonin gene-related peptide (CGRP), and substance P (SP), using radioimmunoassays. In 6 anesthetized horses, colonic ischemia (20% of baseline (BL]) was maintained for 3 hours, then blood flow was restored an...
McKeever KH, Malinowski K.Six young (mean + s.e., 5.3 +/- 0.8 years, 445 +/- 13 kg bwt) and 6 old (22.0 +/- 0.4 years, 473 +/- 18 kg bwt) Standardbred and Thoroughbred mares were used to test the hypothesis that age would alter the endocrine response to exercise. All of the mares were unconditioned but accustomed to the laboratory, to standing quietly and running on a treadmill, and to the standardised incremental exercise test (SET) used in the experiment. Two weeks prior to the experiment, each horse underwent a SET to determine maximal oxygen uptake (VO2max) and the speeds to be used in the actual experiment. A seco...
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...
Clark B, Steel C, Vokes J, Shan JR, Gedye K, Lovett A, Sykes BW.Rebound gastric hyperacidity (RGH) secondary to hypergastrinemia has been suggested to contribute to the rapid recurrence of equine squamous gastric disease (ESGD) in horses after discontinuation of omeprazole. Objective: To evaluate changes in serum gastrin and chromogranin A (CgA) concentrations in response to medium-term (57-day) omeprazole treatment and after omeprazole discontinuation. Methods: Fourteen mature Thoroughbred racehorses in simulated race training. Methods: Horses received 2.28 g of oral omeprazole PO q24h for 57 days within a 61-day period, excluding a withholding period...
Khalil AM, Nakahara K, Tokuriki M, Kaseda Y, Murakami N.On Misaki peninsula, Japan, fecal samples were collected from 14 Misaki stallions at monthly intervals for 12 consecutive months. The fecal testosterone concentration was measured by radioimmunoassay. We examined monthly fecal testosterone hormone patterns and the relationship between fecal testosterone concentration and breeding season and later harem size. Marked monthly variations in fecal testosterone concentration were observed. The fecal testosterone concentration began rising in March; the highest mean monthly concentration, 2.87 +/- 0.18 ng/g, was found in April, and the level remained...
McKeever KH, Antas LA, Kearns CF.The objective of the present study was to measure plasma endothelin-1 (ET-1) at rest and during exercise in the horse. Six healthy, Standardbred and Thoroughbred mares (5.3+/-0.8 years; 445.2+/-13.1 kg) which were unfit, but otherwise accustomed to running on the treadmill, were used in the study. Plasma ET-1 concentrations were measured using a commercially available radioimmunoassay kit. Horses performed three trials: a standing control (CON) trial where blood was collected from the jugular vein every minute for 5 min; a graded exercise test (GXT) where blood samples were collected at the en...
Lemazurier E, Toquet MP, Fortier G, Séralini GE.We used radioimmunoassay (RIA) to measure monthly serum levels of unconjugated and conjugated sex steroids (testosterone T, androstenedione A, estradiol E(2), and estrone E(1)) in 4 male and 4 female foals during their first year of life. Maximal production of sex steroids was detected from April to August with hormonal peaks, corresponding to the natural breeding season in adults. In males, only A levels were more steady. Total estrogens (unconjugated plus conjugated E(2) and E(1)) were the major steroids in immature males in contrast to adults. Estrogens generally peaked in young females bef...
Bailly-Chouriberry L, Chu-Van E, Pinel G, Garcia P, Popot MA, André-Fontaine G, Bonnaire Y, Le Bizec B.Since the Australian commercialisation of the recombinant equine growth hormone (reGH) in 1998 (EquiGen-5), Bresagen), this reGH, which differs only from eGH by an additional methionine at the N-terminal end (met-eGH), is worldwide suspected to be administered to racehorses as a doping agent. Indeed, the use of this biological drug is considered as a threat to horseracing since it acts both on growth, development or reproductive functions, and on the improvement of performances. In this work, we describe two reliable techniques based on surface plasmon resonance biosensor immunoassay (SPR-BIA)...
Haddad NS, Pedersoli WM, Ravis WR, Fazeli MH, Carson RL.Healthy mature pony mares (n = 6) were given a single dose of gentamicin (5 mg/kg of body weight) IV or IM 8 days apart. Venous blood samples were collected at 0, 5, 10, 20, 30, and 45 minutes and at 1, 1.5, 2, 2.5, 3, 4, 6, 8, 10, 12, 18, 24, 30, 36, 40, and 48 hours after IV injection of gentamicin, and at 10, 20, 30, and 45 minutes and at 1, 1.5, 2, 2.5, 3, 4, 6, 8, 10, 12, 18, 24, and 30 hours after IM injection of gentamicin. Gentamicin serum concentration was determined by a liquid-phase radioimmunoassay. The combined data of IV and IM treatments were analyzed by a nonlinear least-square...
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...
Tang PW, Crone DL.A new method for hydrolyzing steroid conjugates (both sulfates and glucuronides conjugates) that is efficient, effective, and inexpensive is described. This method comprises incubation of the conjugates--after salting-out into ethyl acetate or elution from a C18 cartridge--with anhydrous methanolic hydrogen chloride (methanolysis) for 10 min. It has been successfully applied to our routine radioimmunoassay screening and GC/MS confirmation studies of steroids in prerace and postrace equine urine samples. Comparative GC/MS studies on entire (male horse) urine samples showed that methanolysis gav...
Jöchle W, Irvine CH, Alexander SL, Newby TJ.Nine mares received cannulae to collect blood from the pituitary venous outflow in the intercavernous sinus (ICS) and the jugular vein; in 4 mares, only jugular cannulae were used. Those 4 mares and 3 of the mares with cannulae in both positions received 7.5 mg luprostiol i.m. and 1 mare with both cannulae was treated with 3.75 mg uprostiol i.v. Blood samples were kept before and after treatment at 2-, 5- or 10-min intervals and concentrations of LH, FSH and GnRH were determined by RIA. Treatments resulted in an immediate sharp rise of LH and FSH in ICS and jugular blood samples within 2-10 mi...
Smyth GB, Young DW, Hammond LS.Gastrin is the only hormone known to stimulate secretion of hydrochloric acid. It also has trophic effects on specific parts of the mucosa of the gastrointestinal tract. Using radioimmunoassay techniques, postprandial serum gastrin and insulin concentrations were measured in six adult horses to establish effects of different diets on gastrin concentrations. Insulin concentrations were measured to provide support to the patterns of gastrin secretion because patterns of insulin secretion were already known. The horses were fed coastal bermuda hay, or twice daily 5 kg of a complete pelleted ratio...
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...
Panzani S, Villani M, McGladdery A, Magri M, Kindahl H, Galeati G, Martino PA, Veronesi MC.Information regarding the plasma hormone profiles of prostaglandins (PGs), cortisol (C), and progesterone (P4) during pathologic processes in newborn foals is scarce. The aim of this study was to determine the plasma concentrations of these hormones in diseased foals (n=40) and healthy at-term foals (n=24) (Equus caballus) during the first 2 weeks of life. Blood samples were collected daily, before any treatment with nonsteroidal drugs in diseased foals, and plasma was analyzed by radioimmunoassay. 15-Ketodihydro-PGF(2alpha) (PGM) was consistently higher in diseased foals than in healthy foals...
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...
Yoon MJ, Roser JF.Localization of IGF-I and IGF-IR were observed in Leydig cells of horses using immunohistochemistry (IHC), suggesting IGF-I may play a role in equine Leydig cell steroidogenesis. Previous studies in other species have indicated that IGF-I increases basal and/or LH/hCG-induced testosterone production. The objectives of this study were to (1) test the synergistic effect of IGF-I on eLH-induced testosterone production in cultured equine Leydig cells and (2) determine if this effect is reproductive stage-dependent. Testes were collected from five pubertal (1.1±0.1 year; 1-1.5 year) and eight post...
Kinsella HM, Hostnik LD, Rings LM, Swink JM, Burns TA, Toribio RE.The endocrine pancreas and hypothalamic-pituitary-adrenal axis (HPAA) are central to energy homeostasis, but information on their dynamics in response to energy challenges in healthy newborn foals is lacking. Objective: To evaluate glucagon, insulin, ACTH, and cortisol response to fasting and carbohydrate administration in healthy foals. Methods: Twenty-two healthy Standardbred foals ≤4 days of age. Methods: Foals were assigned to fasted (n = 6), IV glucose (IVGT; n = 5), PO glucose (OGT; n = 5), and PO lactose (OLT; n = 6) test groups. Blood samples were collected frequently for 210 min...
Toishi Y, Tsunoda N, Nagata SI, Kirisawa R, Nagaoka K, Watanabe G, Yanagawa Y, Katagiri S, Taya K.Testosterone (T) concentration is a useful indicator of reproductive function in male animals. However, T concentration is not usually measured in veterinary clinics, partly due to the unavailability of reliable and rapid assays for animal samples. In this study, a rapid chemiluminescent enzyme immunoassay system (CLEIA system) that was developed for the measurement of T concentration in humans use was validated for stallion blood samples. First, serum T concentrations were measured using the CLEIA system and compared with those measured by a fluoroimmunoassay that has been validated for use i...
Hale J, Hughes K, Hall S, Labens R.Autologous conditioned serum (ACS) is a common intra-articular treatment for osteoarthritis in horses. The objective of this study was to investigate the influence of ACS preparation method on product contamination and concentrations of relevant cytokines and the influence of multiple freeze/thaw cycles. Blood was obtained from 10 healthy Thoroughbred horses and processed in parallel using a commercial and a non-commercial method to obtain ACS. Fluorescent microsphere immunoassay (FMIA) analysis was performed to quantify Interleukin 1 receptor antagonist (IL-1Ra), Interleukin-10 (IL-10), Inter...
Ferreira CSC, Morais RCL, de Andrade ABP, Balaro MFA, Ribas JAS, Gomes GM, Pinna AE.The aim of this study was to evaluate whether the RI and PI values would help in choosing the best embryo recipient, and observe whether CL vascularization would influence P4 production. During the breeding season 2018/2019, the study was conducted using 35 mares, which is used for reference to collect data for the project on the day of embryo transfer. The utilized mares were divided into five groups followed by the day after ovulation, with D0 being the day of ovulation. Therefore, the five groups are as follows: D4-mares that were on the 4th post-ovulation day; D5-mares that were on the 5th...
Meyers-Brown GA, Loud MC, Hyland JC, Roser JF.The most common equine breeding practice to decrease the time to the first ovulation of the year is to use artificial lights starting December 1 in the Northern Hemisphere. It can take 60-90 d for this lighting regimen to induce a fertile ovulation. The success rate for pharmaceutical compounds to carry out the same process has been variable. One compound that did induce an early ovulation was recombinant equine follicle stimulating hormone (reFSH), but neither pregnancy nor cyclicity was established in that study. Starting on December 1, 20 deep-anestrous mares of light horse breeds (4-15 y...
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...
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...
Ryan PL, Christiansen DL, Hopper RM, Bagnell CA, Vaala WE, Leblanc MM.Placental insufficiency is regarded as the primary factor contributing to late-term abortion and perinatal death of foals. Often when problems associated with late-term pregnancy in the horse are manifest the condition is well-advanced and therapeutic intervention may not be effective in rescuing the pregnancy. If a compromised pregnancy due to placental insufficiency could be identified early, the pregnancy might be sustained through medical intervention. Because the placenta is the sole source of circulating relaxin in the mare, we hypothesized that systemic relaxin may serve as a biomarker ...
Jablonka-Shariff A, Roser JF, Bousfield GR, Wolfe MW, Sibley LE, Colgin M, Boime I.To study structure-activity relationships and the role of equine gonadotropins in the normal and pathophysiology of equine reproduction, the availability of purified hormones is essential. Previous expression studies in transfected CHO cells showed inefficient assembly of the human and bovine alpha and beta subunits, resulting in low levels of recombinant LH. The ability to express a single chain bearing genetically linked alpha and beta subunits bypasses this rate-limiting assembly step. A chimera was constructed by overlap PCR in which the carboxy terminal end of the eLHbeta subunit was gene...
Go YY, Hazard NW, Balasuriya UBR, Chapman AM, Fitton NS, Kenéz Á, Andrews FM.Accurate quantitative analysis of equine insulin in blood samples is critical for assessing hyperinsulinemia in horses. Although there are various laboratory methods for evaluating equine serum insulin, different immunoassays show significant discrepancies between the determined insulin concentrations and are often not comparable. The aim of this study was to evaluate the Immulite® 1000 chemiluminescent immunoassay (CLIA) to establish independent laboratory and assay-specific cut values to provide an accurate diagnosis of hyperinsulinemia in horses. Thus, the analytical and clinical performan...
Saad B, Corradin G, Bosshard HR.The antigenic determinants for two monoclonal antibodies directed against horse apo-cytochrome c, a protein of disordered structure, as judged by spectroscopic and hydrodynamic criteria, have been studied by a combination of methods: antigen competition in solution by radio immunoassay and enzyme-linked immunoassay, and differential acetylation of free and antibody-bound antigen. In the latter method the accessibility of lysine residues of the antigen in the antigen-antibody complex is compared to the accessibility in the free antigen. The two antibodies against the heme-free protein do not re...
