Reproduction in horses encompasses the biological processes and mechanisms involved in the breeding and development of equine offspring. This includes the study of reproductive anatomy, physiology, and endocrinology in both mares and stallions. Key areas of interest include the estrous cycle, ovulation, conception, gestation, and parturition. Researchers also examine factors influencing fertility, reproductive technologies such as artificial insemination and embryo transfer, and management practices that impact reproductive success. This page compiles peer-reviewed research studies and scholarly articles that explore the physiological, genetic, and environmental aspects of equine reproduction.
Vagnoni KE, Ginther OJ, Lunn DP.Chorionic girdle cells are a highly invasive subpopulation of trophoblastic cells of the horse conceptus that adhere to the uterine epithelium and begin to invade the endometrium on Days 34-36 (Day 0 = day of ovulation). Just prior to and during invasion (Days 32-36), chorionic girdle cells express high levels of major histocompatibility complex (MHC) I, but expression of this antigen decreases by Days 40-45 and is lost by Day 55. The mechanisms involved in the control of chorionic girdle cell invasion and altered MHCI expression over time are not known. The objective of this study, therefore,...
Heitland AV, Jasko DJ, Squires EL, Graham JK, Pickett BW, Hamilton C.Five experiments were conducted to evaluate damage incurred in each processing step for cryopreservation of stallion spermatozoa. In Experiment 1, semen was centrifuged for 9 centrifugation times and the percentage of spermatozoa recovered after each treatment was calculated and spermatozoal motion characteristics analysed. Recovery of spermatozoa was > or = 80% when spermatozoa were centrifuged for > or = 10 min. Experiment 2 evaluated spermatozoa cryopreserved at 5 different concentrations in each of 2 extenders (skim milk-egg yolk-glycerol, SM-EYG; and lactose-EDTA, LAC). In SM-EYG, T...
Huhtinen M, Reilas T, Katila T.The pregnancy rate is lower in mares inseminated at the first post-partum (p.p.) oestrus (40-50%) compared with pregnancy rates in subsequent oestrous cycles (55-65%). The causes of the lowered pregnancy rate are not fully understood. The aim of the present study was to examine if embryonic defects could be one of the reasons for lowered pregnancy rate. A total of 23 p.p. and 14 non-lactating control mares were flushed 7 days after detection of ovulation. Embryo recovery rate was 48% and 71% in p.p. and control mares, respectively (p = 0.16). Embryos were photographed, measured, graded and sta...
Nambo Y, Nagata S, Oikawa M, Yoshihara T, Tsunoda N, Kohsaka T, Taniyama H, Watanabe G, Taya K.Plasma concentrations of immunoreactive (ir)-inhibin were measured in seven pregnant mares from around Day 140 of gestation to Day 2 after parturition using a heterologous bovine-based radioimmunoassay (RIA). Concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), oestradiol-17 beta, progesterone and relaxin were also measured in the same samples. A marked increase in plasma concentrations of ir-inhibin, FSH and LH occurred between Day 220 and Day 300 of gestation but the concentrations of all three hormones returned to baseline by about Day 320 (three weeks before part...
Bleumink-Pluym NM, ter Laak EA, Houwers DJ, van der Zeijst BA.The ability of Taylorella equigenitalis, the causative agent of contagious equine metritis, to invade and replicate in equine derm cells was studied. The kinetics of invasion and replication were determined for four T. equigenitalis strains. On the basis of these experiments, a simpler assay in which the invasive as well as the replicative properties of a particular strain could be determined was developed. This assay was used to characterize 32 strains, which had previously been typed by field inversion gel electrophoresis of genomic restriction fragments. The invasiveness of T. equigenitalis...
Thomas PG, Ball BA.To facilitate the study of interactions between equine spermatozoa and homologous oviduct epithelial cells, we developed an assay to count labelled spermatozoa bound to oviduct epithelial cell (OEC) monolayers and used the assay to compare the binding ability of spermatozoa from different stallions. Washed spermatozoa from three stallions were incubated with the fluorochrome Hoechst 33342 (5 micrograms/ml) for 1 min. Spermatozoa were then layered over confluent monolayers of oviduct epithelial cells in 2 cm2 culture wells. Coculture treatments comprised five concentrations of spermatozoa (10(5...
Koskinen E, Huhtinen M, Katila T.Weekly blood samples were collected for serum progesterone determination from 68 mares during the period from September 1990 to June 1991. A total of 78% (53/68) of the mares became anoestrous: 88% (15/17) of mares with foals and 75% (38/51) of dry mares (p = 0.323). Six mares of the 15 which continued cycling showed persistent corpus luteum for 77 days on average (range 42-106 days). Anoestrus began between September and February, most frequently in December, and ended in January to June, most frequently in March. Mares with foal at foot became anoestrous in 2 separate groups, in September an...
Huhtinen M, Reilas T, Katila T.The pregnancy rate is lower in mares inseminated at the first post-partum (p.p.) oestrus (40-50%) compared with pregnancy rates in subsequent oestrous cycles (55-65%). The causes of the lowered pregnancy rate are not fully understood. The aim of the present study was to examine if embryonic defects could be one of the reasons for lowered pregnancy rate. A total of 23 p.p. and 14 non-lactating control mares were flushed 7 days after detection of ovulation. Embryo recovery rate was 48% and 71% in p.p. and control mares, respectively (p = 0.16). Embryos were photographed, measured, graded and sta...
Engelhardt H, King GJ.The epitheliochorial placenta represents the least intimate association between maternal and fetal tissues. The best known examples of this form of placentation are the domestic livestock species. Current information on the nature and proposed functions of uterine lymphocyte populations in ruminants (sheep and cattle), horses and pigs is presented. In ruminants unusual gamma delta T cells may play a role in mid to late gestation. During normal horse pregnancy, fetally derived endometrial cup cells invade the uterine stroma and are destroyed by maternal leukocytes midway through gestation. Natu...
Turner RM, Love CC, McDonnell SM, Sweeney RW, Twitchell ED, Habecker PL, Reilly LK, Pozor MA, Kenney RM.An 8-year-old stallion was evaluated because of recurrent urinary tract infections and chronic intermittent urospermia. After extensive diagnostic testing, it was hypothesized that the stallion had a reflex dyssynergia of the bladder and urethral sphincter. Initial attempts to manage the urospermia included semen fractionation, semen collection after voluntary urination, and use of semen extenders. None of these efforts reliably yielded a quality ejaculate. Administration of imipramine hydrochloride (1.2 mg/kg of body weight, PO, 4 hours prior to semen collection) was initiated in an attempt t...
Broad TE, Forrest JW, Lewis PE, Pearce PD, Phua SH, Pugh PA, Stewart-Scott IA.A DNA repeat element, revealed initially by digestion of horse DNA with TaqI, was cloned and characterized by Southern and in situ hybridization studies and nucleotide sequencing. The clone, e4/1, consisted of 32 tandem reiteration of a unit repeat of 21-22 bp, and produced multilocus DNA fingerprinting profiles that were useful for parentage analysis in horses. The tandem repeat element was shown by in situ hybridization to be localized in the centromeres of the acrocentric but not metacentric classes of horse chromosomes.
Moslemi S, Silberzahn P, Gaillard JL.19-Norandrostenedione was synthesized in vitro from dehydroepiandrosterone by explants of equine full-term placenta. The synthesis of 19-norandrostenedione was inhibited by two specific aromatase inhibitors, 4-hydroxyandrostenedione and fadrozole.
Daels PF, Mohammed HO, Odensvik K, Kindahl H.To determine the relative role of endogenous prostaglandin F2 alpha (PGF2 alpha) secretion in cloprostenol-induced abortion in mares that no longer require luteal progesterone secretion for maintenance of pregnancy, and to evaluate the ability of a prostaglandin cyclooxygenase inhibitor (flunixin meglumine) to prevent cloprostenol-induced abortion. Methods: The effect of flunixin meglumine on PGF2 alpha secretion and outcome of pregnancy was compared between mares treated with cloprostenol only and mares treated with cloprostenol plus flunixin meglumine. Methods: Five pregnant mares, aged 4 to...
Thomas PG, Ignotz GG, Ball BA, Brinsko SP, Currie WB.Adhesion of equine spermatozoa to homologous oviduct epithelial cells (OEC) in vitro results in specific changes in spermatozoa and OEC function. To test the hypothesis that adhesion of spermatozoa affects protein synthesis and secretion by OEC, the following treatment groups were established in culture: OEC with culture medium only; control spermatozoa in culture medium only; OEC in coculture with spermatozoa; and OEC and spermatozoa in coculture, but physically separated by a microporous membrane. The experiment was replicated within each of 4 ejaculates from 3 stallions. De novo protein sec...
Paweska JT, Volkmann DH, Barnard BJ, Chirnside ED.Two in a group of five naturally seropositive donkey stallions were found to shed equine arteritis virus (EAV) in their semen as demonstrated by virus isolation. Direct intramuscular inoculation of sonicated semen from one virus-shedding stallion (S3) caused clinical disease in two donkeys from which virus was recovered and in which seroconversion was detected. Sexual transmission was confirmed in two mares mated to S3 when after a febrile response during which EAV was isolated from huffy coats and nasal and ocular exudates, both mares were found to have seroconverted. In-contact transmission ...
Simonsen U, Prieto D, Sánez de Tejada I, García-Sacristán A.1. The involvement of nitric oxide (NO) and the signal transduction mechanisms mediating neurogenic relaxations were investigated in deep intracavernous penile arteries with an internal lumen diameter of 600-900 microns, isolated from the corpus cavernosum of young horses. 2. The presence of nitric oxide synthase (NOS)-positive nerves was examined in cross and longitudinal sections of isolated penile arteries processed for NADPH-diaphorase (NADPH-d) histochemistry. NADPH-d-positive nerve fibres were observed in the adventitia-media junction of deep penile arteries and in relation to the trabec...
Ijaz A, Ducharme R.Stallion semen was diluted in five different extenders (dimitro-poulus onze (Dimitro's), Kenney's modified tryode (Kenney's), modified INRA82 (INRA82), egg yolk-citrate-taurine (Citrate) and EZ-Mixin) and evaluated for motility after cooling and storage at 5 degrees C for 0, 24, 48, 72 and 96 h. EZ-Mixin extender was used as control while 70 and 100 mM of taurine were added to Dimitro's, Kenney's and INRA82 to study its effect under conditions of storage at 5 degrees C and varying processing modifications. Motility in INRA82 was 57.0, 58.4, 61.1, and 56.1% after 24, 48, 72 and 96 h, respective...
Allen WR, Mathias S, Lennard SN, Greenwood RE.Rapid enzyme-based immunoassays were used to measure concentrations of oestradiol-17 beta and progesterone in daily blood samples recovered throughout oestrus and for a few days after ovulation from 34 Thoroughbred and 8 pony-type maiden, barren and foaling mares. The first detectable fall in oestradiol-17 beta levels occurred in 88% of the mares within the interval -72 to 0 h with respect to ovulation and in 65% of mares within the interval of -48 to 0 h. The results indicated that serial daily hormone assays of this type could, in a high proportion of animals, predict a correct time for a si...
Re G, Badino P, Novelli A, Di Renzo GF, Severino L, De Liguoro M, Ferone MR.The distribution of oestrogen and progesterone receptors in the equine genital tract was investigated by means of a modified dextran-coated charcoal method on samples collected from the vagina, the cervix and the uterus of 30 healthy adult Polish mares, divided into two groups on the basis of their serum progesterone levels. The concentrations of oestrogen and progesterone receptors were significantly (P < 0.05) lower in the vagina and the cervix than in the uterus, in agreement with data from human beings, cattle and pigs, which showed that the highest concentrations of oestrogen and proge...
Brinsko SP, Ball BA, Ignotz GG, Thomas PG, Currie WB, Ellington JE.The time of activation of the embryonic genome (maternal-embryonic transition) in equine embryos was investigated by assessing incorporation of 3H-uridine and nucleolar development. In Experiment 1, embryos were recovered from the oviduct (n = 15) and the uterus (n = 3). Recovered embryos were assessed for morphologic development and quality score. Recovered embryos with less than 8 cells (two cells, n = 4; four cells, n = 5; five cells, n = 2) were incubated with 3H-uridine (560 microCi/ml) for 10 hr, while eight-cell embryos (n = 2), morulae (n = 2), and blastocysts (n = 3) were incubated wi...
Aurich C, Daels PF, Ball BA, Aurich JE.The aim of the present study was to investigate the role of ovarian steroids in the opioid regulation of LH and prolactin release in mares. Effects of the opioid antagonist naloxone on LH and prolactin secretion were determined in ovariectomized pony mares. The animals were pretreated with either progesterone (500 micrograms kg-1) or oestradiol benzoate (10 micrograms kg-1) for 8 days and subsequently with a combination of progesterone and oestradiol for an additional 8 days. Naloxone administration (0.5 mg kg-1 i.v.) resulted in a significant release of LH as well as prolactin in mares after ...
Gastal EL, Kot K, Ginther OJ.A technique for intrafollicular treatment with a transvaginal ultrasound-guided injection needle was developed using equine chorionic gonadotropin (eCG) as the test substance. An injection was made into one growing follicle of a wave when the follicles were 20 to 23 mm. The treated follicles were injected with 1000 iu of eCG in 0.2 ml saline solution and control follicles were injected with 0.2 ml of the saline vehicle (10 mares per group, 1 follicle per mare). The injection system used an inner 25-gauge needle and an outer 20-gauge needle inserted together through the needle-guide channel of ...
Hinshelwood MM, Liu Z, Conley AJ, Simpson ER.Conversion of androgens to estrogens is catalyzed by aromatase P450 (P450arom; the product of the CYP19 gene). Regulation of tissue-specific expression of P450arom in humans is due, in part, to alternative transcriptional start sites that arise as a consequence of the use of granulosa cells and placental tissue from cows, horses, and pigs (ungulates) in order to determine whether these species, like the human, utilize tissue-specific promoters to drive P450arom expression. The majority of transcripts in the placenta have 5'-termini that differ from those in the ovary upstream of a common site ...
Dobrinski I, Thomas PG, Ball BA.Two bioassays were used to evaluate the interaction of fresh and cryopreserved equine semen with oviductal epithelial cells (OEC) and with the zona pellucida (ZP). Split ejaculates were either stored at room temperature or frozen and thawed. In experiment 1, progressive motility and membrane integrity were evaluated for each treatment. Fluorescent labeled spermatozoa were cocultured with monolayers of OEC for 30 minutes, and the number of sperm attached to OEC was counted by fluorescence microscopy and analysis of digitized images. Motility of spermatozoa attached to OEC was observed at 0.5, 3...
Lennard SN, Stewart F, Allen WR.In situ hybridization, Northern blotting, and immunohistochemical techniques were used to study the expression of transforming growth factor beta 1 (TGF beta 1) in the endometrium of the mare during the first 150 days of pregnancy (term = 330-340 days). In situ hybridization using an oligonucleotide (45mer) probe, based on a homologous region within all known mammalian TGF beta 1 DNA sequences, demonstrated TGF beta 1 mRNA accumulation in the glandular and lumenal epithelial cells of the endometrium from day 33 onwards which corresponds to the time of implantation (day 33-45). Expression in th...
Glatzel PS, Belz JP.Because of the mare's ability to conceive very soon post partum a thoroughly performed control of the puerperal period is of particular importance. In addition to cytological and histological examinations of the uterus, rectal palpation and vaginoscopic inspection as well as microbiological examinations of uterine swabs and the evaluation of the mare's hormonal status during the puerperal period provide useful information which can be used either prognostically or to initiate a specific therapy. 55 mares were examined on days 3, 6 and 9 post partum. Rectal palpation, vaginoscopic inspection, m...
Long MT, Goetz TE, Kakoma I, Whiteley HE, Lock TE, Holland CJ, Foreman JH, Baker GJ.Fetal infectivity of Ehrlichia risticii was investigated in 19 ponies that were E risticii negative on the basis of results of an indirect fluorescent antibody (IFA) test. Thirteen pregnant ponies were infected by IV administration of E risticii between 90 and 180 days of gestation. Six pregnant ponies served as noninfected controls. Each infected pony had clinical signs of equine monocytic ehrlichiosis, was confirmed to be ehrlichemic, and developed an IFA titer to E risticii. Two infected ponies became recumbent, were unresponsive to supportive care, and were euthanatized. After recovery fro...
Peippo J, Huhtinen M, Kotilainen T.A rapid and reliable method for sex determination of preimplantation-stage equine embryos has not been available. The aim of the present study was to find an enzyme which would distinguish sexes in the horse by finding a polymorphic restriction site between the ZFY and ZFX homologues amplified by the polymerase chain reaction (PCR). Altogether, 38 different restriction enzymes were tested using female and male DNA extracted from blood. The primers used for amplification were selected from conserved sequences between human ZFY and ZFX genes and mouse Zfy-1 and Zfy-2 genes. Nine enzymes cut the ...
Fazeli AR, Steenweg W, Bevers MM, van den Broek J, Bracher V, Parlevliet J, Colenbrander B.The hemizona assay (HZA) has been developed as a diagnostic test to predict the fertilisation potential of human spermatozoa. The aim of this study was to develop an HZA for stallion spermatozoa and to investigate a possible relationship between fertility and the outcome of the HZA in this species. Equine oocytes were obtained from ovaries collected at a slaughterhouse and by transvaginal, ultrasound-guided follicle aspiration. They were then denuded from cumulus cells and stored in salt solution at 4 degrees C until use. On the day of the experiments the oocytes were bisected, thus providing ...
Silva DM, Holden SA, Lyons A, Souza JC, Fair S.The aim of the present study was to assess the effect of the addition of docosahexaenoic acid (DHA) on the in vitro quality of cooled and frozen-thawed stallion semen. In Experiment 1, semen from 10 stallions was collected (three ejaculates per stallion). Semen was diluted to 100×106 spermatozoa mL-1 with 0.02mM vitamin E (VE) and 0, 1, 10 or 20ng mL-1 DHA and frozen. Semen was thawed and total motility (TM), rapid progressive motility (PM), acrosome integrity, membrane fluidity and morphology were assessed. In Experiment 2, semen from three stallions was collected (three ejaculates per stall...
Ginther OJ, Griffin PG.The extent and nature of fetal mobility (presentation and location changes) were studied in 10 pony mares by ultrasonic examinations each week during the fetal stage (Day 40 to term). The percentage of examinations with cranial fetal presentation was 35 to 43% during Months 2 to 5 (no significant differences among months); a significant increase occurred between Months 5 and 6 reaching 100% at Month 7. With only 3 transient exceptions, the fetus was in cranial presentation during all examinations from Month 7 to term. The frequency of presentation changes between successive examinations decrea...
Basrur PK, Kanagawa H, Podliachouk L.An intersex horse exhibiting cell types of different sex chromosome constitution was subjected to further studies in order to determine whether the house was a mosaic or a chimera. Cultures of gonadal tissue and peripheral blood revealed mainly 64/XX and 64/XY cells, the former predominating in both tissues. The frequency of drumstick-bearing poly-morphonuclear neutrophils in the intersex horse was similar to that noted in normal mares. Blood type analysis using 17 naturally occurring agglutinins and hemolysins revealed partial agglutinations with three antibodies for the factors of the A syst...
Long MT, Goetz TE, Kakoma I, Whiteley HE, Lock TE, Holland CJ, Foreman JH, Baker GJ.Fetal infectivity of Ehrlichia risticii was investigated in 19 ponies that were E risticii negative on the basis of results of an indirect fluorescent antibody (IFA) test. Thirteen pregnant ponies were infected by IV administration of E risticii between 90 and 180 days of gestation. Six pregnant ponies served as noninfected controls. Each infected pony had clinical signs of equine monocytic ehrlichiosis, was confirmed to be ehrlichemic, and developed an IFA titer to E risticii. Two infected ponies became recumbent, were unresponsive to supportive care, and were euthanatized. After recovery fro...
Fitzgerald BP, Reedy SE, Sessions DR, Powell DM, McManus CJ.The seasonal nature of reproductive activity in mares is widely accepted and considerable attention has focused on the mechanisms that lead to the initiation of the breeding season. In contrast, considerably less information is available about the termination of the breeding season. It is interesting to note that each winter a sub-population of mares continues to undergo oestrous cyclicity during the non-breeding season. Continuation of reproductive activity during the winter occurs most frequently in mares that maintain a non-pregnant condition in successive years. The maintenance of a non-pr...
Moussa M, Bersinger I, Doligez P, Guignot F, Duchamp G, Vidament M, Mermillod P, Bruyas JF.Vitrification using open pulled straw (OPS) has provided encouraging results with embryos from other species. The aim of this study was to compare the survival of 6.5- and 6.75-day-old equine embryos after OPS vitrification and slow-cooling. Eighteen embryos were frozen using a slow-cooling method. Embryos were placed in modified PBS with increasing glycerol concentration (2.5%, 5%, 7.5% and 10% (v/v) 5 min each). Embryos were loaded into 0.25 ml straws then placed in a programmable freezer and subsequently plunged into liquid nitrogen. After thawing, cryoprotectant was removed by five steps w...
Quartuccio M, Marino G, Mannarino C, Cristarella S.Congenital abnormalities of the uterus and the cervix are rare in horses, often associated with infertility, despite normal ovarian activity and sexual behaviour. An abnormality of the cervix in a 5-year-old jennet with a history of infertility was described. Clinical and ultrasound examination of the genital tract showed a normal development of the uterus with regular cyclic activity in both the ovaries. Vaginoscopy revealed a normal development of the vagina and a cervix deviated on the left side of the vagina by a thick dorsoventral fold. The lateral deviation was permanent in any stage of ...
Fernandes CB, Loux SC, Scoggin KE, Squires EL, Troedsson MH, Esteller-Vico A, Ball BA.The cervix is a dynamic structure that undergoes dramatic changes during the estrous cycle, pregnancy and parturition. It is well established that hormonal changes, including estrogens, progestogens and prostaglandins, regulate the expression of key proteins involved in cervical function. The arachidonic acid cascade is important in the remodeling and relaxation of the cervix in the days preceding parturition. Despite the complexity of this mechanism, regulation of cervical function has received little study in the mare. Therefore, the objective of this study was to compare the expression of e...
Jellyman JK, Valenzuela OA, Allen VL, Holdstock NB, Fowden AL.Pancreatic β cells are responsive to a range of stimuli during early post natal life in healthy pony foals. However, little is known about whether these responses are sex-linked. Objective: To determine pancreatic β cell responses to the insulin secretagogues, glucose, arginine and tolbutamide, in fillies and colts during the first 3 months after birth. Methods: In vivo experiment examining sex differences in pancreatic β cell function in foals. Methods: Female (n = 8) and male (n = 5) pony foals were infused i.v. with glucose (0.5 g/kg bwt 40% dextrose), arginine (100 mg/kg bwt) or tol...
Nes N, Lømo OM, Bjerkås I.In 4 female foals of the Norwegian horse breed, "Fjord Horse", congenital arthrogryposis of the limbs are described. The disorder was mainly limited to the hind limbs and associated with polydactylia and partly with brachygnathia superior and cleft palate. The defective foals were paternal halfsisters of 4 normal foals (1 female + 3 males), sired by the stallion "Bingo" 1804, which was phenotypically quite normal. The disorder being lethal, is possibly caused by a sex-limited or a strongly sex-influenced dominant gene.
Newcombe JR, Paccamonti D, Cuervo-Arango J.Data were analysed retrospectively from fourteen breeding seasons at an Equine Fertility Clinic for the effect of interval between pre- and postovulatory examinations for immediate postovulatory insemination on pregnancy rate (PR) and embryo loss rate (ELR). Mares of various breeds and ages were examined at intervals which varied from 0.5 to 15h between the pre- and postovulatory period over 867 cycles. When ovulation was detected they were inseminated with a single dose of commercial frozen-thawed semen. All mares were treated in the post-insemination period with intrauterine antibiotics and ...
Siddiqui MA, Gastal EL, Gastal MO, Beg MA, Ginther OJ.Follicle blood flow, follicular-fluid and plasma hormone concentrations, and oocyte quality were studied 30 h after an ovulation-inducing hCG treatment when the pre-ovulatory follicle was 32 mm. Mares were grouped as positive (n = 16) and negative (n = 44) for hCG antibodies before the experimental hCG treatment. Percentage of the follicle wall with blood flow signals was less (p < 0.05) in the antibody positive group than in the negative group. The concentrations of follicular-fluid oestradiol and free IGF1, and plasma oestradiol were greater (p < 0.001), and follicular-fluid progesterone (p ...
Senra RL, Ramírez-López CJ, Magalhães-Júnior MJ, Neves JGDS, Barros E, Waddington B, Guimarães SEF, Guimarães JD, Baracat-Pereira MC.Horses are seasonal polyoestrous animals, and the photoperiod is the main factor modulating their reproductive activity. There is no consensus on the andrological and biochemical factors that influence breeding seasonality. To assess the involvement of climate in reproduction, Mangalarga Marchador stallions were monitored over 1 year regarding semen quality and seminal plasma proteome. Here, we show that kallikrein (KLKs) proteoforms in seminal plasma are involved in climate conditioning of reproduction. During the breeding season, greater abundance and different types of KLKs occurred simult...
Morris LH, Allen WR.The need for relatively high numbers of spermatozoa for artificial insemination limits our application of recently available technologies such as sex-sorted semen. The fertility of two different methods of low dose insemination using fresh, frozen and sex-sorted semen are compared in this overview. Satisfactory conception rates are described using very low doses of spermatozoa inseminated by either hysteroscopic or deep uterine insemination methods, proving the stallion is fully fertile. The hysteroscopic method appears to give higher conception rates when inseminating fewer than 5 x 10(6) spe...
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...
McCue PM, Fleury JJ, Denniston DJ, Graham JK, Squires EL.A technique was developed for oviductal insemination of mares, in which a small number of motile spermatozoa are deposited directly into the oviduct. Pregnancy rates in mares inseminated by traditional intrauterine artificial insemination were compared with rates in mares inseminated by oviductal insemination. Fifteen mares were inseminated with 5 x 10(8) progressively motile spermatozoa by intrauterine artificial insemination, and 14 mares were inseminated with 5 x 10(4) progressively motile spermatozoa by oviductal insemination. Pregnancy rates in mares inseminated by intrauterine artificial...
Neves AP, Keller A, Trein CR, Möller G, Jobim MI, Castilho LF, Cardoso MR, Leibold W, Zerbe H, Klug E, Gregory RM, Mattos RC.This study compared four treatments for bacterial endometritis in mares experimentally infected with Streptococcus zooepidemicus. Twenty-five mares were used, 20 resistant and five susceptible to endometritis. Mares would be in estrus when infected. Twenty-four hours after inoculation, clinical, bacteriological and cytological examinations were performed and repeated until the first occurrence: negative cytology and no Streptococcus growth or the seventh day post-infection. All mares showed clinical signs of endometritis and were assigned to one of the following treatments: (1) intrauterine in...
Townsend J, Westcott K, Tortonese DJ.In the horse, pronounced changes in fertility occur annually in response to photoperiod. However, the mechanisms regulating gonadotrophin synthesis and release in this species remain unclear. Here, we investigated the expression of gonadotrophin subunits and GnRH receptor (GnRH-R) mRNA in the pituitary glands of Thoroughbred horses during the breeding (BS) and non-breeding (NBS) season. Seasonal effects on the prevalence of gonadotrophs in the pars distalis were also examined. GnRH-R and common alpha-, LHbeta- and FSHbeta-subunit mRNA contents were determined by Northern analysis and the preva...
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...
Marshall DE, Dumasia MC, Wooding P, Gower DB, Houghton E.Maternal endometrial and fetal allantochorionic tissues were separated manually from the placentae of seven healthy thoroughbred and three pony mares, ranging in gestational age from 100 to 318 days. The homogeneity of subcellular fractions prepared from these tissues was assessed initially using the marker enzymes, succinate dehydrogenase, NADPH cytochrome C reductase and lactate dehydrogenase for the mitochondrial, microsomal and cytosolic fractions, respectively. Light microscopy and histochemical analysis demonstrated that the separated fetal allantochorionic membrane, which is made up of ...
Viafara JAS, Portilho RV, Maculan R, Souza Júnior JF, Silva CM, Rodrigues PG, El Azzi MS, Souza JC.This study evaluated the presence of polyunsaturated fatty acids in circulating blood and in the ovarian follicular fluid of mares, after supplementation of the diet with linseed oil. Six Mangalarga Marchador mares, weighing 397.00±31.89 kg, were kept on native pasture, and assigned to the current study. In a switch over design, mares were randomly allocated to receive 150 ml of vegetable oil daily, containing polyunsaturated fatty acids n3 (62.23 g ALA, 20.34 g LA, 2.27 g EPA, 2.32 g DHA), (n=3) or no supplementation (n=3) in two replicates. Blood and follicular fluid samples were taken on t...
Pruß D, Oldenhof H, Wolkers WF, Sieme H.The aim of this study was to establish an alginate encapsulation procedure for stallion sperm, and investigate if sperm encapsulation enhances longevity during cold storage and survival after cryopreservation. First, biocompatibility of the compounds needed for encapsulation was tested and factors determining capsule structure were identified. Sperm encapsulation was realized either by depositing droplets (20 µL) of sperm solution supplemented with barium or calcium chloride (10 mM) in alginate solution (0.25%, w/v), or by adding sperm-alginate droplets in solution containing barium or calc...
Dongus H, Beelitz P, Schöl H.The female reproductive system of Thelazia lacrymalis (Nematoda: Thelaziidae) was investigated by light and scanning electron microscopy (SEM) with regard to the developmental stages and the stage deposited by the gravid nematode. Female T. lacrymalis have a didelphic and opisthodelphic type of reproductive system with paired ovaries, oviducts and uteri and a single vagina and vulva. Round and spindle-shaped primary oocytes are documented within the ovaries and oviducts, respectively. The distal part of each uterus provides a fertilization chamber filled with spermatozoa, followed by a sphinct...
De Lille AJ, Silvers ML, Cadario ME, Tran TQ, Cage CL, LeBlanc MM.The effects of the sedatives xylazine (predominantly alpha2 agonist) and acepromazine (predominantly alpha1 antagonist) and their influence on intrauterine pressure were measured in four reproductively normal mares and four mares with delayed uterine clearance. Intrauterine pressure was measured in each mare during oestrus for three 100 min periods using an 8-F catheter with three discrete pressure sensors. After 10 min of baseline recording, xylazine, acepromazine or saline was administered i.v. Oxytocin was administered i.v. after a further 30 min, and pressure recordings were continued for ...
McDonnell SM, Love CC, Martin BB, Reef VB, Kenney RM.Two mature breeding stallions were evaluated because of specific ejaculatory dysfunction, and each was found to have aortic-iliac thrombosis occluding 60 to 70% of the aortic lumen. In each case, the stallion had strong libido, normal mounting, and vigorous initial thrusting. With continued exertion, thrusting became weak and dismount was awkward. Treatment aimed at maximizing sexual arousal before mounting and reducing hind limb pain and exertion during breeding allowed these stallions to continue breeding. A program of gradually increasing daily exercise also was associated with improved bre...
Adeyefa CA.The effects of the various growth factors with regard to the nutritional physiology of zoophilic dermatophytes isolated from horses are reported. The optimum temperature, pH and growth period for the fungal isolates were found to be 30 degrees-35 degrees C, 5-6 and 7-12 days respectively while the carbon and nitrogen sources utilised by the organisms were sucrose, fructose, maltose, L-arginine and calcium nitrate. The use of these factors in preparation of efficacious fungicides used in the treatment of ringworm infections in man and animals is discussed.
McDonnell SM, Oristaglio Turner RM.Imipramine-induced ex copula ejaculates (11) and fractionated in copula ejaculates were collected from each of 5 pony stallions for freezing in 5-ml straws (6), using a modified Kenney glucose skim-milk extender (2). Initial post-thaw total and progressive motilities and daily post-thaw total and progressive motilities, as well as the number of days to reach 0 progressively motile spermatozoa, were also similar for the 2 methods of collection. The percentage of morphologically normal spermatozoa both before freezing and after thawing were also similar for in copula and ex copula ejaculates. Co...
Lea RG, Stewart F, Allen WR, Ohno I, Clark DA.Endometrial tissue from the gravid uterine horn of pregnant mares was examined by northern analysis and in situ hybridization for mRNA that hybridized to cDNA and RNA probes generated from a mouse TGF-beta 2 1.2 kb cDNA clone. The mouse cDNA probe hybridized to characteristic TGF-beta 2 mRNA transcripts on a northern blot of total RNA isolated from horse endometrium collected at day 45 of gestation. Two major 4.0 and 3.5 kb transcripts and possibly a minor 1.6 kb transcript were observed, consistent with specific hybridization to equine TGF-beta 2 mRNA. By in situ hybridization, riboprobes tra...
Hansen PJ, Asbury AC.The release of opsonins into the uterine lumen of mares susceptible or resistant to endometritis was examined after intrauterine inoculation of a filtrate of Streptococcus culture fluid or vehicle. Uterine flushings were collected at 0.5 hour before and 2, 4, 6, 8, and 24 hours after inoculation on day 2 or 3 of estrus and on day 7 or 8 after ovulation. Amounts of opsonins in flushings were quantified as the H2O2 produced by leukocytes incubated with flushings-opsonized bacteria, compared with H2O2 produced by leukocytes incubated with nonopsonized bacteria. Opsonin values in flushings increas...
