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Topic:Experimental Methods

Experimental methods in equine research encompass a variety of techniques and procedures used to investigate physiological, behavioral, and health-related aspects of horses. These methods are designed to collect data that can enhance the understanding of equine biology and inform veterinary practices. Common experimental approaches include controlled trials, observational studies, and laboratory-based experiments. Techniques may involve the use of non-invasive imaging, biochemical assays, and physiological monitoring to assess parameters such as metabolic function, locomotion, and disease progression. This page compiles peer-reviewed research studies and scholarly articles that explore the application, development, and evaluation of experimental methods in the study of equine science.
In vitro development of day-2 equine embryos co-cultured with oviductal explants or trophoblastic vesicles.
Theriogenology    March 1, 1991   Volume 35, Issue 3 669-682 doi: 10.1016/0093-691x(91)90462-m
Ball BA, Altschul M, Ellington JE.This study compared the in vitro development of Day-2 equine embryos co-cultured with either trophoblastic vesicles or oviductal explants. Embryos were collected surgically from the oviducts of pony mares 2 d after ovulation and assessed for stage of development. Culture medium was Ham's F12 and Dulbecco's Modified Eagle's Medium (50:50 v/v) in a humidified atmosphere of 5% CO(2) in air at 38.5 degrees C with either trophoblastic vesicles or oviductal explants. The quality score of embryos was assessed daily. After 4 d in culture, embryos were stained (Hoechst 33342) and evaluated with epifluo...
[Year-round antibody profile of groups of horses of a herd kept in isolation after differently terminating use of an experimental viral combination vaccine].
DTW. Deutsche tierarztliche Wochenschrift    March 1, 1991   Volume 98, Issue 3 82-89 
Bürki F, Nowotny N, Hofer A.The commercial vaccine "Resequin F Konz." devised against viral respiratory infections of horses contains the abortigenic Equine Herpesvirus-1 (EHV-1). Therefore we had used it in our protection project of the Austrian Lipizzaners+ primarily to prevent abortions. Taking into account the recent perception that for young horses the respiratory-pathogenic EHV-4 type is essential Behringwerke Marburg added this particular virus to their market product to produce a multicomponent experimental vaccine. We examined this vaccine for its antibody induction as well as their persistence against each of i...
Role of antibody to extracellular proteins of Rhodococcus equi in protection against R. equi pneumonia in foals.
Veterinary microbiology    February 15, 1991   Volume 26, Issue 4 323-333 doi: 10.1016/0378-1135(91)90026-c
Machang'u RS, Prescott JF.Rhodococcus equi produces two exoenzymes (REE), a cholesterol oxidase in large amounts and a phospholipase C, which cause lysis of sheep red blood cells (SRBC) sensitized with Staphylococcus aureus beta toxin. Two immunization studies were done in foals to determine the role of antibody to REE in protection against R. equi pneumonia. In the first study, three foals (mean age 10 days) were vaccinated four times at 2-week intervals with over 1 million units of partially purified exoenzymes (PREE). In the second study, three foals (mean age 19 days) were administered plasma from an adult horse va...
Effects of hydroxyflutamide on rats treated with a superovulatory dose of pregnant mare serum gonadotropin.
Canadian journal of physiology and pharmacology    February 1, 1991   Volume 69, Issue 2 185-190 doi: 10.1139/y91-027
Yu FH, Yun YW, Yuen BH, Moon YS.Immature female rats treated with superovulatory doses of pregnant mare serum gonadotropin (PMSG) were used to study the effects of the antiandrogen hydroxyflutamide on steroid production, particularly the biologically active androgens, in two experiments. In the first experiment, animals were given either 5 mg hydroxyflutamide or vehicle alone at 30 and 36 h following 40 IU PMSG. Compared with the vehicle group, hydroxyflutamide treatment significantly reduced the percentage of degenerate oocytes recovered from oviducts (p less than 0.05). Serum levels of testosterone and androstenedione, and...
Evaluation of progesterone deficiency as a cause of fetal death in mares with experimentally induced endotoxemia.
American journal of veterinary research    February 1, 1991   Volume 52, Issue 2 282-288 
Daels PF, Stabenfeldt GH, Hughes JP, Odensvik K, Kindahl H.The role of decreased luteal activity in embryonic loss after induced endotoxemia was studied in mares 21 to 35 days pregnant. Fourteen pregnant mares were treated daily with 44 mg of altrenogest to compensate for the loss of endogenous progesterone secretion caused by prostaglandin F2 alpha (PGF2 alpha) synthesis and release following intravenous administration of Salmonella typhimurium endotoxin. Altrenogest was administered daily from the day of endotoxin injection until day 40 of gestation (group 1; n = 7), until day 70 (group 2; n = 5), or until day 50 (group 3; n = 2). In all mares, secr...
Biochemical study of repair of induced osteochondral defects of the distal portion of the radial carpal bone in horses by use of periosteal autografts.
American journal of veterinary research    February 1, 1991   Volume 52, Issue 2 328-332 
Vachon AM, McIlwraith CW, Keeley FW.Periosteal autografts were used for repair of large osteochondral defects in 10 horses aged 2 to 3 years old. In each horse, osteochondral defects measuring 1.0 x 1.0 cm2 were induced bilaterally on the distal articular surface of each radial carpal bone. Control and experimental defects were drilled. Periosteum was harvested from the proximal portion of the tibia and was glued into the principal defects, using a fibrin adhesive. Control defects were glued, but were not grafted. Sixteen weeks after the grafting procedure, the quality of the repair tissue of control and grafted defects was asse...
Pulsed radio frequency therapy of experimentally induced arthritis in ponies. Crawford WH, Houge JC, Neirby DT, Di Mino A, Di Mino AA.The effect of pulsed radio frequency therapy (PRFT) was evaluated on seven ponies with no arthritis and in 28 ponies in which arthritis was created using intra-articular amphotericin B to induce synovitis in the right middle carpal joint. The ponies were divided into five treatment and two control groups. Two levels of arthritis were created and two dosage levels of PRFT were evaluated. The effect of PRFT on arthritic and nonarthritic joints was measured by comparing synovial fluid parameters, the degree and duration of lameness, the range of carpal motion, and carpus circumference, for treate...
The corpus luteum: source of oestrogen during early pregnancy in the mare.
Journal of reproduction and fertility. Supplement    January 1, 1991   Volume 44 501-508 
Daels PF, DeMoraes JJ, Stabenfeldt GH, Hughes JP, Lasley BL.Thirty pregnant mares were assigned to 3 groups: Group 1 (n = 10) mares served as controls; Group 2 (n = 10) mares were treated with altrenogest (44 mg/day) from Day 16 to 80 and Group 3 (n = 10) mares were treated with a luteolytic dose of PGF2 alpha on Day 16 followed by altrenogest (44 mg/day) until Day 80. Concentrations of progesterone and chorionic gonadotrophin (CG) in plasma and oestrogen conjugate (OC) in urine were determined between Days 16 and 80 of gestation. In Group 3, complete luteolysis occurred in all 10 mares following administration of PGF2 alpha. Six of the 10 mares did no...
A preliminary study of immunological castration in colts.
Journal of reproduction and fertility. Supplement    January 1, 1991   Volume 44 183-190 
Dowsett KF, Pattie WA, Knott LM, Jackson AE, Hoskinson RM, Rigby RP, Moss BA.This study tested the effectiveness of a conjugated GnRH vaccine for stimulating antibody production, suppressing testosterone secretion and depressing testicular development in yearling colts. Two colts were allocated to each of three groups, (1) control, (2) subcutaneous and (3) intramuscular vaccinations. Two injections of the vaccine were given 11 weeks apart. Liveweight gain was not affected by vaccination but plasma testosterone concentrations in the treated colts were suppressed and their antibody titres to GnRH were greater than 1:1000. Testicular development in the treated colts also ...
Effect of interval between doses on response of the pony to sodium bicarbonate.
The Cornell veterinarian    January 1, 1991   Volume 81, Issue 1 59-66 
Kowalski J, Roberts A, Williams J, Hintz HF, Daniluk P, Schryver HF.Three pony geldings were given sodium bicarbonate orally in order to study the effect on blood pH and bicarbonate and to determine if frequency of dosing influences the response. In a preliminary study, it appeared that a carry-over effect might occur if the interval between dosing was only 2 days. The ponies received 2 doses of sodium bicarbonate (400 mg/kg) 7 days apart in trial one and then in trial two they received 2 doses of sodium bicarbonate 4 days apart. The sodium bicarbonate was mixed with 2 liters of warm water and given through a nasogastric tube on each trial day. Blood samples w...
Effect of constant light exposure on circulating gonadotrophin levels and hypothalamic gonadotrophin-releasing hormone (GnRH) content in the ovariectomized pony mare.
Journal of reproduction and fertility. Supplement    January 1, 1991   Volume 44 259-266 
Cleaver BD, Grubaugh WR, Davis SD, Sheerin PC, Franklin KJ, Sharp DC.Melatonin is thought to play a role in relaying photic information to the central nervous system as part of the seasonal reproductive cycle of the mare. However, the mechanisms by which melatonin may act are unknown. Therefore, this study was designed to determine whether exposure to constant light would, by reducing circulating melatonin concentrations, have any effect on hypothalamic gonadotrophin-releasing hormone (GnRH) content and circulating levels of luteinizing hormone (LH) and follicle-stimulating hormone (FSH). Blood samples were collected for 12 h at 15-min intervals from 8 ovariect...
Establishment of equine oviduct cell monolayers for co-culture with early equine embryos.
Journal of reproduction and fertility. Supplement    January 1, 1991   Volume 44 393-403 
Battut I, Bézard J, Palmer E.A culture for equine oviduct epithelial cells is described. Primary cultures reached confluence in 5-8 days, forming a monolayer of polygonal cells and remaining morphologically intact for about 20 days. Subcultures were obtained by collecting cells detached spontaneously from the monolayers, and confluence was reached again after 5-7 days. Cells frozen before primary culture were confluent 10-15 days after thawing. Dishes containing confluent cells also were frozen, and some cohesive monolayers formed after thawing. Equine embryos, collected 2 days after ovulation, were cultured alone or with...
Experimental models of endotoxaemia related to abortion in the mare.
Journal of reproduction and fertility. Supplement    January 1, 1991   Volume 44 509-516 
Kindahl H, Daels P, Odensvik K, Daunt D, Fredricksson G, Stabenfeldt G, Hughes JP.Three different routes of administering Salmonella typhimurium endotoxin to mimic naturally occurring endotoxaemia were tried in the mare. Bolus injection, repeated bolus injections and continuous low-dose infusion were compared with prostaglandin F2 alpha release, leucocyte count and clinical response. A biphasic prostaglandin release and a pronounced leucopenia of almost identical patterns were seen in all models. Repeated bolus injections showed that the second injection initiated only a small prostaglandin release indicating the development of refractoriness to the treatment. A similar ref...
Experimental cutaneous free flap transfers in the horse.
Microsurgery    January 1, 1991   Volume 12, Issue 2 130-135 doi: 10.1002/micr.1920120214
Lees MJ, Fretz PB, Bowen CV, Leach DH.Equine limb wounds often heal slowly by epithelialization, and large scars are a frequent end result. In some ways, they resemble the wound associated with human tibial injuries. The literature indicates that previous investigators have failed to transfer free skin flaps successfully in the horse. In this paper, we review our experimental work with the deep circumflex iliac flap in the horse. Dissections of 20 cadavers confirmed the anatomical consistency of the flap. Four flaps survived well when elevated as island flaps, but five orthotopic and nine heterotopic free flap transfers all failed...
Effect of oral melatonin on the date of the first ovulation after ovarian inactivity in mares under artificial photoperiod.
Journal of reproduction and fertility. Supplement    January 1, 1991   Volume 44 249-257 
Guillaume D, Palmer E.Two experiments tested the hypothesis that exogenous melatonin (12 mg) given 4 h before dusk (evening melatonin), or near dawn (morning melatonin), would mimic a prolongation of the night and suppress stimulation of the ovaries induced by long days. Experiment 1 consisted of a non-stimulated control group, a control group stimulated by 14.5 h of light, a treated group stimulated with 14.5 h of light plus evening melatonin, a control group given 17.5 h of light and a treatment group given 17.5 h of light plus morning melatonin. The mean (+/- s.e.m.) intervals from the start of treatment to the ...
Effects of human alpha interferon on experimentally induced equine herpesvirus-1 infection in horses.
American journal of veterinary research    December 1, 1990   Volume 51, Issue 12 2006-2010 
Seahorn TL, Carter GK, Martens JG, Crandell RA, Martin MT, Scrutchfield WL, Cummins JM, Martens RJ.The immunotherapeutic effect of low-dose human alpha interferon on viral shedding and clinical disease was evaluated in horses inoculated with equine herpesvirus-1 (EHV-1). Eighteen clinically healthy weanling horses, 5 to 7 months old, were allotted to 3 equal groups. Two groups were treated orally with human alpha-2a interferon (0.22 or 2.2 U/kg of body weight), on days 2 and 1 before inoculation with EHV-1, the day of inoculation, and again on postinoculation day 1. The horses of the remaining group were given a placebo orally on the same days. The horses were monitored daily for changes in...
Effect of endotoxin administration on equine digital hemodynamics and starling forces.
American journal of veterinary research    November 1, 1990   Volume 51, Issue 11 1703-1707 
Hunt RJ, Allen D, Moore JN.Using a pump-perfused extracorporeal isolated digital preparation, the effects of a 30-minute infusion of either saline solution (control) or endotoxin on equine digital hemodynamics and microvascular function were determined. Digital blood flow and arterial, venous, and capillary pressures were recorded at 15-minute intervals for 150 minutes. From these data, total vascular resistance and pre- and postcapillary resistances were calculated. Isogravimetric capillary filtration coefficient, vascular compliance, and the osmotic reflection coefficient were determined after the last hemodynamic mea...
Left ventricular volume determination in the horse by two-dimensional echocardiography: an in vitro study.
Equine veterinary journal    November 1, 1990   Volume 22, Issue 6 398-402 doi: 10.1111/j.2042-3306.1990.tb04303.x
Vörös K, Holmes JR, Gibbs C.This study was designed to evaluate the accuracy of two-dimensional echocardiography (2DE) in determining the left ventricular volume (LVV) of the horse in vitro. After examining the shape of the left ventricular silicon rubber casts of four equine hearts, two modified Simpson's rule methods (Model A and Model B) as combinations of conical shapes and one biplane area-length method as a single cone (Model C) were chosen for volume calculations. One long axis and three short axis planes were used for linear and area 2DE measurements, respectively. The ventricular length (L) was calculated from t...
Cultivation of tissue from the matrix of the stratum medium of the equine and bovine hoof walls.
American journal of veterinary research    November 1, 1990   Volume 51, Issue 11 1852-1856 
Ekfalck A, Rodriguez-Martinez H, Obel N.Explants from the matrix of the stratum medium of the wall of the equine and bovine hoof each were cultured on a microporous membrane, using a standard culture medium. After incubation at 37 C, the outgrowth was a mixture of keratinocytes and fibroblasts, with predominance of the latter. After incubation at 34 C, the keratinocytes dominated, covering the lateral surfaces of the explant as well as the basal surface. Lateral outgrowth of keratinocytes was observed at the borderline of the original epidermis and at the borderline of the explant's contact with the membrane. Epithelial outgrowth fr...
Effects of active immunization against GnRH on LH, FSH and prolactin storage, secretion and response to their secretagogues in pony geldings.
Journal of animal science    October 1, 1990   Volume 68, Issue 10 3322-3329 doi: 10.2527/1990.68103322x
Rabb MH, Thompson DL, Barry BE, Colborn DR, Hehnke KE, Garza F.Six pony geldings were actively immunized against GnRH conjugated to bovine serum albumin (BSA) to study 1) the relative dependency of LH and FSH storage, secretion and response to GnRH analog on GnRH bioavailability and 2) the effects of reduced GnRH bioavailability on GnRH storage in the hypothalamus. Five geldings were immunized against BSA. Geldings were immunized in December and 4, 8, 14, 20, 26 and 32 wk later. Ponies immunized against GnRH had increased (P less than .01) GnRH binding in plasma within 6 wk. By June, plasma concentrations of LH and FSH in ponies immunized against GnRH had...
Phagocytosis of opsonized fluorescent microspheres by equine polymorphonuclear leukocytes.
Zentralblatt fur Veterinarmedizin. Reihe B. Journal of veterinary medicine. Series B    September 1, 1990   Volume 37, Issue 7 481-490 doi: 10.1111/j.1439-0450.1990.tb01087.x
Foerster RJ, Wolf G.Equine blood polymorphonuclear leukocytes (PMN) were isolated by buffy coat and hypotonic lysis of residual erythrocytes. A highly reproducible method is described for measuring the uptake of opsonized latex microspheres by equine PMN using flowcytometry. The use of cytochalasin D allowed for differentiation of ingested from attached particles. The kinetics of phagocytosis in vitro is shown for different experimental conditions. We developed an assay for evaluation of phagocytic capacity of PMN which allows the assessment of drugs for their influence on phagocytosis in vivo as well as in vitro...
Questions experimental design in study on effects of furosemide on racing times of Thoroughbreds.
American journal of veterinary research    September 1, 1990   Volume 51, Issue 9 1505-1506 
Vulliet R.No abstract available
Attempted Ehrlichia risticii transmission with Dermacentor variabilis (Acari: Ixodidae).
Journal of medical entomology    September 1, 1990   Volume 27, Issue 5 931-933 doi: 10.1093/jmedent/27.5.931
Levine JF, Levy MG, Nicholson WL, Gager RB.Larval Dermacentor variabilis (Say) (n = 327) were fed on Balb/C mice inoculated with Ehrlichia risticii, the etiologic agent of equine monocytic ehrlichiosis (Potomac horse fever). All mice displayed clinical signs of E. risticii infection at the time of feeding. After molting, resulting nymphs (n = 74) were fed on susceptible mice. No clinical signs were observed, and the mice remained seronegative for 6 wk after feeding.
Serum lipid and lipoprotein changes in ponies with experimentally induced liver disease.
American journal of veterinary research    September 1, 1990   Volume 51, Issue 9 1380-1384 
Bauer JE, Meyer DJ, Campbell M, McMurphy R.Alterations in serum lipid and lipoprotein concentrations in ponies with experimentally induced liver disease were investigated. Hepatocellular damage was induced, using a nonlethal dose of carbon tetrachloride. In a separate group of ponies, obstructive jaundice was induced by surgical ligation of the common bile duct. Over a 6-day period, blood samples were obtained from ponies after treatment with carbon tetrachloride and for 12 days in ponies subjected to surgery. Serum cholesterol and triglyceride concentrations were unaffected in both groups of ponies, except for significantly (P less th...
Horner’s syndrome in the horse: a clinical, experimental and morphological study.
Equine veterinary journal. Supplement    September 1, 1990   Issue 10 62-65 doi: 10.1111/j.2042-3306.1990.tb04714.x
Simoens P, Lauwers H, De Muelenare C, Muylle E, Steenhaut M.A clinical case of Horner's syndrome is described in a Standardbred horse, and the various symptoms of cranial sympathetic denervation are studied in two ponies after experimental transection of the left cervical sympathetic trunk and vagosympathetic trunk, respectively. The most prominent symptoms of equine Horner's syndrome were ptosis, local sweating and increased cutaneous temperature in the denervated area. Enophthalmos, miosis and increased lacrimation were also observed but these symptoms were mild, variable and difficult to ascertain. Prolapse of the third eyelid was not noticed. Conco...
Attempted transmission of Ehrlichia risticii (Rickettsiaceae) with Stomoxys calcitrans (Diptera: Muscidae).
Journal of medical entomology    September 1, 1990   Volume 27, Issue 5 874-877 doi: 10.1093/jmedent/27.5.874
Burg JG, Roberts AW, Williams NM, Powell DG, Knapp FW.Experimental transmission of Ehrlichia risticii, the causal agent of Potomac horse fever, was attempted with adult stable flies, Stomoxys calcitrans, (L.) using two feeding schedules. In schedule A, a set of 140 flies was allowed to feed once on an experimentally infected donor pony and once 24 h later on a recipient pony. A different set of flies was used each day for a 12-d period. In schedule B, 240 flies were allowed to feed once daily for 12 consecutive d on the donor pony followed by five consecutive daily feedings on the recipient pony. E. risticii was isolated from the blood of the exp...
Synovial fluid pH, cytologic characteristics, and gentamicin concentration after intra-articular administration of the drug in an experimental model of infectious arthritis in horses.
American journal of veterinary research    September 1, 1990   Volume 51, Issue 9 1363-1369 
Lloyd KC, Stover SM, Pascoe JR, Adams P.Chemical and cytologic effects and bactericidal activity of gentamicin in septic synovial fluid were evaluated in an experimental model of infectious arthritis in horses. Septic arthritis was induced by inoculation of approximately 7.5 X 10(6) colony-forming units of Escherichia coli into 1 antebrachiocarpal joint in each of 16 clinically normal adult horses. Clinical signs of septic arthritis were evident 24 hours after inoculation. Horses were allotted to 3 groups: group-1 horses (n = 5) each were given 150 mg of gentamicin (50 mg/ml; 3 ml) intra-articularly (IA); group-2 horses (n = 5) each...
The influence of photoperiod on gonadotrophin-releasing hormone stimulated luteinising hormone release in the anoestrous mare.
Equine veterinary journal    September 1, 1990   Volume 22, Issue 5 356-358 doi: 10.1111/j.2042-3306.1990.tb04289.x
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 ...
Ultrastructural morphology of hypomelanosis in equine cutaneous papilloma.
Journal of comparative pathology    August 1, 1990   Volume 103, Issue 2 199-213 doi: 10.1016/s0021-9975(08)80176-4
Hamada M, Itakura C.The morphology of hypomelanosis occurring in experimentally induced equine papillomas was investigated. Histologically, dopa-positive functioning melanocytes were decreased in number from the basal layer in the epidermis. Electron-microscopically, melanogenic organelles in the melanocytes were degenerate and melanosomes were decreased in number and size. In addition, the melanocytes had some abnormal melanosomes including melanosome complexes and giant melanosomes. Some abnormal melanosomes were also present in the keratinocytes. The hypomelanosis seemed to be related to a disturbance in melan...
Effects of clustered drill holes on the breaking strength of the equine third metacarpal bone.
American journal of veterinary research    August 1, 1990   Volume 51, Issue 8 1242-1246 
Specht TE, Miller GJ, Colahan PT.The breaking strength (stress at failure) of equine third metacarpal bones, with and without clustered drill holes, was determined in vitro. Paired ossa metacarpalia II-IV of 39 horses (n = 39) between 2 and 7 years old were tested in palmarodorsal 3-point bending. Four treatments were compared. Clustered 2.7- or 3.5-mm drill holes, in a 4- or 7-hole pattern, were made in the dorsal cortex of the distal diaphysis of the left third metacarpal bone. Undrilled right third metacarpi were used as controls. Bones with clustered drill holes failed by an oblique fracture through 1 or more drill holes,...
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