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.
Camacho Benítez A, Vasconcellos R, Lombide P, Viotti H, Pérez W, Cazales N, Cavestany D, Martin GB, Pedrana G.Heat shock proteins play a crucial role in cellular development, proliferation, differentiation and apoptosis. Heat shock protein 90 (HSP90) has been localised in the human endometrium, where its immunoexpression changes during the menstrual cycle. Similar studies have not been done for the equid species, so the present study aimed to describe endometrial HSP90 immunoexpression in mare endometrium. Endometrial biopsies were formalin-fixed and paraffin-embedded, and sections were stained with haematoxylin-eosin in preparation for HSP90 immunohistochemistry. Immunostaining and morphometric analy...
Shilton CA, Kahler A, Davis BW, Crabtree JR, Crowhurst J, McGladdery AJ, Wathes DC, Raudsepp T, de Mestre AM.The first 8 weeks of pregnancy is a critical time, with the majority of pregnancy losses occurring during this period. Abnormal chromosome number (aneuploidy) is a common finding in human miscarriage, yet is rarely reported in domestic animals. Equine early pregnancy loss (EPL) has no diagnosis in over 80% of cases. The aim of this study was to characterise aneuploidies associated with equine EPL. Genomic DNA from clinical cases of spontaneous miscarriage (EPLs; 14-65 days of gestation) and healthy control placentae (various gestational ages) were assessed using a high density genotyping arr...
Cavalero TMS, Segabinazzi LGTM, Scheeren VFC, Canuto LEF, Gobato MLM, Papa FF.A 7-year-old Quarter Horse stallion was admitted at the hospital with a history of ejaculatory failure for 12 months. The stallion revealed no physical or psychological abnormalities, as well as, normal libido and erection. In addition, there were no abnormalities in accessory sex glands or the aorta artery detected by transrectal ultrasonography. Based on clinical findings, the stallion was diagnosed with an idiopathic ejaculatory dysfunction; therefore, alternative attempts of semen collection were performed. Thermal compress on the basis of the stallion's penis, semen collection on the gro...
Campos GA, Garcia VFC, Freitas-Dell'Aqua CP, Segabinazzi LGTM, Maciel LFS, Alvarenga MA, Papa FO, Dell'Aqua JA.This study aimed to assess the effects of sodium caseinate and cholesterol to extenders used for stallion semen cooling. Two ejaculates from 19 stallions were extended to 50 million/mL in four different extenders and cooled-stored for 24 hours at 5°C. The extender 1 (E1) consisted of a commercially available skim milk-based extender. The extender 2 (E2) consisted of E1 basic formula with the milk component being replaced by sodium caseinate (20 g/L). The extender 3 (E3) consisted of E1 basic formula added to cholesterol (1.5 mg/120 million sperm). The extender 4 (E4) consisted of a combinat...
Perdomo-González DI, Sánchez-Guerrero MJ, Molina A, Valera M.The aim of this work was to analyze genetic parameters such as the inbreeding coefficient (F), relatedness coefficient (AR) and partial inbreeding coefficient (Fij) of the whole PRE population, and the ancestors which account for 50% of the total genetic variability of the current population, from genealogical information. The average F of the whole PRE population (328,706 animals) has decreased from 8.45% to 7.51% in the least 20 years. The Fij was estimated for the whole PRE population, resulting in a database of 58,772,533 records containing one record for each Fij that each animal receives...
Treulen F, Aguila L, Arias ME, Jofré I, Felmer R.In vitro manipulation of spermatozoa leads to deleterious changes of structure and function that occur mainly due to oxidative stress, therefore, prevention or treatment is a strategy to improve the functions of processed sperm. In the present study, the aim was to evaluate the effects of MnTBAP supplementation, a compound with antioxidant activity, on in vitro capacitation conditions of thawed equine sperm. For this purpose, stallion spermatozoa (2 × 10 cells/mL) were incubated in the sperm-TLP base medium for 4 h in which there were three different conditions: non-capacitating, capacitating...
Todd ET, Thomson PC, Hamilton NA, Ang RA, Lindgren G, Viklund Å, Eriksson S, Mikko S, Strand E, Velie BD.Domestic animal populations are often characterised by high rates of inbreeding and low effective population sizes due to selective breeding practices. These practices can result in otherwise rare recessive deleterious alleles drifting to high frequencies, resulting in reduced fertility rates. This study aimed to identify potential recessive lethal haplotypes in the Thoroughbred horse breed, a closed population that has been selectively bred for racing performance. In this study, we identified a haplotype in the LY49B gene that shows strong evidence of being homozygous lethal, despite having h...
Immunoglobulin type G (IgG) concentration both in jennies' colostrum and in serum of donkey foals are mostly unknown in the first 24 h after delivery. The aims of the present study were to evaluate the IgG concentrations of colostrum during the first 24 h of lactation of Amiata jennies, the absorption of colostrum and the weekly body weight gain of the donkey foals. IgG concentrations were assessed in the jennies' colostrum and in the serum of donkey foals. Colostrum was collected in 9 jennies ready after delivery, and at 6, 12, 24 h after foaling from both halves. Serum was collected at the s...
Al-Kass Z, Guo Y, Vinnere Pettersson O, Niazi A, Morrell JM.Bacteria colonize stallion semen during collection and processing which may cause disease in inseminated females or negatively affect sperm quality during storage prior to insemination. Antibiotics are added to semen extenders to control the growth of these bacteria but may induce antimicrobial resistance. Research into alternatives to antibiotics for this purpose requires knowledge of which bacteria are present in semen. Not all bacteria in semen, however, can be identified by conventional microbiological techniques. The objectives of the study were to: i) determine which bacteria are present...
Günay Uçmak Z, Kurban I, Uçmak M.Endometritis is a major cause of infertility in mares. The aim of this study is to evaluate the preovulatory follicle (POF) vascularization (A mix, A red, A blue), POF diameter, POF wall thickness, and uterine diameters in mares with or without endometritis. Ten healthy mares and 10 mares with endometritis diagnosed by the combination of transrectal palpation, ultrasonographic examination, and cytology brush were enrolled in the study. Data of the groups obtained at 2 days before the ovulation (day -2) were compared with Student's t-test. Correlations of the parameters were determined by the ...
Magee C, Bruemmer JE, Kirkley KS, Sylvester LA, Runyan B, Nett TM, Squires EL, Clay CM.To more clearly understand the equine gonadotrope response to kisspeptin and gonadotropin releasing hormone (GnRH), peripheral LH and FSH were quantified in diestrous mares after treatment with either equine kisspeptide (eKp-10, 0.5 mg iv), GnRH (25 μg iv), or a combination thereof every 4 h for 3 days. The following observations were made: 1) a diminished LH and FSH response to eKp-10 and GnRH was observed by Day 3, but was not different by treatment, 2) a decrease in basal LH concentration was observed from Day 1 to Day 3 for the eKp-10, but not the GnRH treated mares, 3) there was no ch...
Oliveira SN, Andrade LRP, Silva LFMC, Araujo EAB, Rayashi RM, Segabinazzi LGTM, Alvarenga MA, Dell'Aqua CP, Dell'Aqua JA, Papa FO.Treatments for seminal vesiculitis have poor outcomes in stallions; thus, the development of alternative strategies is warranted. This study aimed to evaluate fractionated semen collection as a method to restore the fertility of stallions diagnosed with seminal vesiculitis. Eighteen ejaculates from six stallions (three ejaculates/stallion) diagnosed with seminal vesiculitis were harvested in fractions, as follows: Fraction A (FA), the first two jets; Fraction B (FB), the third and fourth jets; and Fraction C (FC), the fifth and remaining jets of the ejaculate. All fractions were subject to sta...
Vissani MA, Perglione CO, Zabal O, Alvarez G, Thiry E, Barrandeguy M, Parreño V.Equid alphaherpesvirus 3 (EHV-3) is the etiological agent of equine coital exanthema (ECE). Because no vaccines or antiviral therapies are available, prevention consists of clinical examination of mares and stallions before mating or semen collection and resting from breeding activities when lesions are present. However, this methodology does not identify subclinically infected animals. Ganciclovir is the most potent compound known to reduce EHV-3 replication. This study aimed to evaluate the efficacy of topical ganciclovir application to reduce EHV-3 replication in experimentally infected mar...
Rakic R, Bourdon B, Demoor M, Maddens S, Saulnier N, Galéra P.An amendment to this paper has been published and can be accessed via a link at the top of the paper.
Dini P, Bartels T, Revah I, Claes AN, Stout TAE, Daels P.A high degree of inbreeding has been reported to negatively impact semen quality in Friesian horses and Shetland ponies. Both breeds are characterized by a closed studbook, small population size, and high incidence of inbreeding. The Dutch Warmblood studbook (KWPN: Koninklijk Warmblood Paardenstamboek Nederland) is a much larger studbook with two distinct populations: the KWPN-Riding horses, managed as an 'open' studbook, and the KWPN-Harness horses, representing a much smaller subpopulation within the KWPN breed and managed as an 'almost closed' studbook. It was recently reported that the deg...
Toishi Y, Tsunoda N, Kirisawa R.In 2017, two Thoroughbred stallions, A and B in Farms A and B, respectively, in Hokkaido in Japan showed clinical signs of equine coital exanthema (ECE). In 2020, stallion C in Farm B showed clinical signs of ECE. Eighteen mares were mated within five days before stallion A developed ECE. Ten mares that mated within 3 days before onset showed clinical signs of ECE on the external genitalia. Equine herpesvirus 3 (EHV-3) was isolated from vaginal swabs from three mares that mated within 2 days before onset. Swabs from 12 mares that mated within 4 days before onset were real-time PCR (rPCR)-posit...
Wilsher S, Rigali F, Kovacsy S, Allen WT.Vitrification of embryos >300 µm in diameter requires puncture of the glycoprotein capsule, although the size of the hole compatible with embryo survival is unknown. Forty-five day-7 or -8 embryos were punctured using a 30-µm glass biopsy pipette mounted on a micromanipulator (n = 20) or manually with either an acupuncture needle (∼100-µm diameter -hole; n = 10) or a microneedle with a <1 µm tip to produce a ∼30-µm diameter hole (n = 15) before transferring to recipient mares; further 12 embryos were punctured with either the acupuncture needle or microneedle before being cultured ...
Ponthier J, de la Rebière G, Salciccia A, Deleuze S.In mares, endometrial cysts are associated with endometriosis and can cause maternal recognition failure or compromise and delay pregnancy diagnoses. Historical treatments were invasive and had adverse effects on the endometrium. Hysteroscopically guided laser therapy is easy and effective for endometrial cysts resection, with no deleterious effects for the endometrium. A 110 cm long and 1.0 cm wide endoscope is sterilely introduced in the uterus through the open cervix of an estrous mare after vulvar cleaning. The uterus is slowly infused with less than 1 L of physiologic solution and the las...
Melgaard DT, Korsgaard TS, Thoefner MS, Petersen MR, Pedersen HG.Unwanted behaviour in mares is a commonly presenting problem to the veterinarian. This behaviour may range from the mare being uncooperative or aggressive when handled on the ground, kicking, bucking or rearing when ridden or being aggressive towards other horses. This purpose of the study was to evaluate whether bilateral ovariectomy in mares with unwanted behaviour improved the mare's behaviour and/or rideability from the owner's perspective. The mares were grouped and compared statistically based on their histological classification as having either "normal" or neoplastic ovaries. This stud...
Edwards L, Finno CJ.Neurologic disease in horses can be particularly challenging to diagnose and treat. These diseases can result in economic losses, emotional distress to owners, and injury to the horse or handlers. To date, there are 5 neurologic diseases caused by known genetic mutations and several more are suspected to be heritable: lethal white foal syndrome, lavender foal syndrome, cerebellar abiotrophy, occipitoatlantoaxial malformation, and Friesian hydrocephalus. Genetic testing allows owners, breeders, and veterinarians to make informed decisions when selecting dams and sires for breeding or deciding t...
Proverbio D, Perego R, Baggiani L, Ferrucci F, Zucca E, Nobile F, Spada E.A knowledge of the blood groups and alloantibodies present is essential for the safe transfusion of blood products in horses. Pre-transfusion screening and blood typing minimizes the risk of incompatible RBC transfusions and prevents immunization of the recipient against incompatible RBC antigens. The frequencies of blood groups can vary among different breeds. Knowledge of a breed's blood group prevalence can be very useful for identifying the best blood donors during transfusion in clinical practice. The aims of this study were to estimate the prevalence of the Ca blood type in horses from I...
Rieger J, Kaessmeyer S, Al Masri S, Hünigen H, Plendl J.The cardiovascular system is the first functional organ in the embryo, and its blood vessels form a widespread conductive network within the organism. Blood vessels develop de novo, by the differentiation of endothelial progenitor cells (vasculogenesis) or by angiogenesis, which is the formation of new blood vessels from existing ones. This review presents an overview of the current knowledge on physiological and pathological angiogenesis in the horse including studies on equine endothelial cells. Principal study fields in equine angiogenesis research were identified: equine endothelial progen...
Cruz A, Sullivan DB, Doty KF, Hess RA, Canisso IF, Reddi PP.The acrosome plays a critical role in sperm-oocyte interactions during fertilization. SP-10 is an acrosomal matrix protein, which is evolutionarily conserved among mammals. The SP-10 antibody has been shown to be useful for staging the seminiferous cycle in the mouse and human. A canonical acrosomal marker; however, has never been used for staging in the horse. The objectives of the present study were to investigate the presence of SP-10 within the horse acrosome using an anti-mouse SP-10 antibody, to classify spermatids based on the shape of the acrosome, and then to use that information to a...
Sinclair CD, Thompson D, Stevenson JS, Rozell T, Jager A, Kouba J.Introducing males to seasonally anestrous females has been shown to increase LH concentrations and hasten the first ovulation in ewes and goat does. Investigation of this sociosexual phenomenon in mares has not been well studied. The objective of this study was to determine if direct or indirect exposure of anestrous mares to a stallion would result in a similar increase in LH and hasten the first ovulation. In late February, mares were stratified by parity and age and assigned randomly to three treatments (n = 7/treatment): (1) DXP (direct exposure; mares housed adjacent to stallion and all...
Gastal EL, Aguiar FLN, Gastal GDA, Alves KA, Alves BG, Figueiredo JR.The mammalian ovary is responsible for essential stages of folliculogenesis and hormonal production, regulating the female physiological functions during the menstrual/estrous cycles. The mare has been considered an attractive model for comparative studies due to the striking similarities shared with women regarding in vivo and in vitro folliculogenesis. The ovarian follicular population in horses contains a large number of oocytes enclosed in preantral follicles that are yet to be explored. Therefore, the in vitro manipulation of equine preantral follicles aims to avoid the process of atre...
De Amicis I, Bucci R, Giangaspero B, D'Alterio N, Trachtman AR, Robbe D.Orchitis and epididymo-orchitis are inflammatory lesions of the testicle. We herein describe a case of monolateral chronic orchitis which occurred in a Tiro Pesante Rapido (TPR) stallion, born in 2002, with a history of good fertility. The stallion was healthy and asymptomatic although the left testis was found to be smaller as compared with the right one and was hard in consistency. Histopathology examination revealed tubular atrophy and parenchymal sclerosis. Scattered foci of calcification and chronic inflammation, the latter dominated by macrophages and lymphocytes, were also observed. Alt...
Herrera MF, Otermin M, Herrera JM, Simoy MV, Bianchi CP, Aguilar JJ, Fumuso EA.Mycobacterium cell wall fraction (MCWF) is a biological component made up of molecules with immunostimulant properties, which is therapeutically used to modulate persistent breeding-induced endometritis (PBIE). The aim of this study was to analyze the effect of this immunomodulator on the endometrial histological structure during the diestrus of PBIE-resistant and -susceptible mares that either received treatment with MCWF or not. The experiment was conducted with 10 resistant mares (RM) and 9 susceptible mares (SM). In the first estrous cycle of the trial, all mares were inseminated with dead...
Alamaary MS, Haron AW, Hiew MWH, Ali M.Present study aimed to investigate the effect of adding antioxidants, cysteine and ascorbic acid on the levels of glutamic oxaloacetic transaminase (GOT), glutamic-pyruvate (GPT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and γ-glutamyl transpeptidase (GGT) enzymes of post-thawed stallion sperm. Ten ejaculates were collected each from four healthy stallions and cryopreserved using HF-20 freezing extender containing either 0 mg/ml cysteine or ascorbic acid, 0.5 mg/ml cysteine and 0.5 mg/ml ascorbic acid. All samples in freezing extender containing cysteine or ascorbic acid or ...
Camacho CA, Estradé MJ, Cazales N, Caballeros JE, Fiala-Rechsteiner SM, Neves AP, Mattos RC.This experiment aimed to verify if the proteins present in a 13 day conceptus induce changes in the equine endometrial ultra-structure, histology, and vascularization, two days after its infusion. Ten healthy cyclic mares were used. Once estrus was confirmed, mares were examined daily to detect ovulation (day 0). After ovulation, mares were examined daily until day seven by transrectal palpation and B-mode and Doppler ultrasonography. In this first cycle, intrauterine biopsies were collected at day seven after ovulation, constituting the Cyclic group (n = 10). In the second cycle, the same mar...
Donkey placenta does not allow the passage of immunoglobulins; thus, foals are born hypogammaglobulinemic and an adequate intake of high-quality colostrum in the first 24 hours of life is crucial for the surviving. The study aims to assess the relation between colostrum immunoglobulin G (IgG) concentration evaluated by the single radial immunodiffusion (SRID) test and the Brix refractometer in donkeys to establish a cutoff value for high quality of colostrum based on Brix refractometry. Colostrum was collected at foaling, and at 6, 12, and 24 hours after foaling from the left and the right h...
Adams GP, Kastelic JP, Bergfelt DR, Ginther OJ.The incidence of intrauterine fluid collections during dioestrus (12/43, 28%) and uterine cysts throughout the oestrous cycle (11/73, 15%) found in this study indicates that these ultrasonically detectable abnormalities are prevalent in mares. The hypothesis that uterine cysts do not affect pregnancy was not supported. Intrauterine fluid collections at dioestrus represented the presence of an inflammatory process as indicated by a high biopsy score, reduced progesterone concentrations, and a shorter interovulatory interval. Mares with fluid collections at dioestrus had a lower pregnancy rate a...
Roberts EL, Dang T, Lepage SIM, Alizadeh AH, Walsh T, Koch TG, Kallos MS.Equine mesenchymal stromal cells (MSCs) are increasingly investigated for their clinical therapeutic utility. Such cell-based treatments can require cell numbers in the millions or billions, with conventional expansion methods using static T-flasks typically inefficient in achieving these cell numbers. Equine cord blood-derived MSCs (eCB-MSCs), are promising cell candidates owing to their capacity for chondrogenic differentiation and immunomodulation. Expansion of eCB-MSCs in stirred suspension bioreactors with microcarriers as an attachment surface has the potential to generate clinically rel...
Ortved KF, Nixon AJ, Mohammed HO, Fortier LA.To evaluate the long-term clinical outcome after allogeneic chondrocyte and insulin-like growth factor-I (IGF-I) grafting of subchondral cystic lesions (SCLs) of the femoral condyle in horses. Objective: To test the hypothesis that chondrocyte and IGF-I grafts will improve the long-term clinical outcome in arthroscopically debrided SCLs. Methods: Medical records of 49 horses with SCLs of the femoral condyle treated by debridement and implantation of chondrocytes and IGF-I were reviewed. Preoperative radiographs were obtained, and caudocranial radiographic projections were used to establish a r...
Schulman ML, Botha AE, Muenscher SB, Annandale CH, Guthrie AJ, Bertschinger HJ.Active immunisation against gonadotrophin-releasing hormone (GnRH) provides a reversible method for control of oestrous behaviour and fertility in mares. Previous reports failed to demonstrate the interval to resumption of cyclic ovarian activity after GnRH-vaccination. Objective: Administration of the GnRH-vaccine Improvac in a large group of mares of various ages will result in effective, reliably reversible suppression of ovarian activity within a 2 year period. Methods: The mares, subdivided into 3 age categories, were vaccinated twice (with a 35 day interval) using 400 µg Improvac and mo...
Wingfield Digby NJ, Ricketts SW.The results of 4024 concurrent endometrial swab and smear tests taken from maiden, barren and post-parturient mares during early oestrus, before coitus demonstrated the practical value of these tests in routine stud farm practice. The use of cytological examinations provided a more direct diagnostic test for acute endometritis and this increased the accuracy of interpretation of the bacteriological findings.
Leblanc MM.Most equine infertility cases can be solved with a methodical, thorough physical and reproductive examination and appropriate diagnostic laboratory aids. Repeated examinations may be needed in some cases to identify subtle anatomical abnormalities or irregularities between hormonal and physiological relationships of the reproductive tract. For pregnancy to occur, hormonal signaling must be exquisitely synchronized with physical changes of the reproductive tract and deposition of fertile semen in the uterus. Asynchrony of these events, infection, inflammation, previous trauma to the reproductiv...
Swierstra EE, Gebauer MR, Pickett BW.The cycle of the seminiferous epithelium of the stallion was
divided into eight stages, using as criteria the presence of meiotic divi-
sions, shape of the spermatid nuclei and location of spermatids with
elongated nuclei in the tubule. The mean frequencies ofstages 1 to 8 were
16\m=.\9, 14\m=.\9, 3\m=.\2, 15\m=.\8, 7\m=.\4, 13\m=.\5, 12\m=.\6 and 15\m=.\7%, respectively. The
duration of one cycle of the seminiferous epithelium was 12\m=.\2 days
(S.E.\m=+-\0\m=.\1) as determined by injecting a single dose of 700 \g=m\Ciof
[3H]thymidine into each spermatic artery of six stallions and rem...
The paper is a report from a workshop discussing equine endometritis, a condition affecting horse fertility. The event was held in honor of Professor John Hughes and his significant contributions […]
Sevinga M, Vrijenhoek T, Hesselinks JW, Barkema HW, Groen AF.This study was motivated by the hypothesis that the incidence of retained placenta (RP) in Friesian horses is associated with inbreeding. The objectives were to 1) calculate the inbreeding rate in the total registered Friesian horse population; 2) study the association of the inbreeding coefficient of the foal and the mare with the incidence of RP; and 3) study the heritability of RP in Friesian mares after normal foalings. Data from the total registered Friesian horse population from 1879 to 2000 (52,392 individuals) were collected from the registration files of the Friesian Horse Studbook. I...
Roach JM, Foote AK, Smith KC, Verheyen KL, de Mestre AM.Pregnancy loss after Day 70 of gestation manifests as abortion, stillbirth or perinatal death. While previous studies have reported the diagnoses of laboratory submissions, none have quantified the incidence and causes of abortions, stillbirths and perinatal mortality at a population level. Objective: To report the incidence and causes of pregnancy loss after Day 70 of gestation in a cohort of Thoroughbreds. Methods: Retrospective cohort study. Methods: Outcomes of Day 70 pregnancies were collected from eight Thoroughbred farms over the 2013-2017 breeding seasons. Stud, veterinary and laborato...
Hall SE, Aitken RJ, Nixon B, Smith ND, Gibb Z.Oxidative stress is a major determinant of mammalian sperm function stimulating lipid peroxidation cascades that culminate in the generation of potentially cytotoxic aldehydes. The aim of this study was to assess the impact of such aldehydes on the functionality of stallion spermatozoa. The impact of exposure to exogenous acrolein (ACR) and 4-hydroxynonenal (4HNE) was manifested in a highly significant dose- and time-dependent increase in mitochondrial reactive oxygen species (ROS), total cellular ROS, a decrease in sperm motility, and a time-dependent increase in lipid peroxidation. Notably, ...
Gaitskell-Phillips G, Martín-Cano FE, Ortiz-Rodríguez JM, Silva-Rodríguez A, Gil MC, Ortega-Ferrusola C, Peña FJ.Although cryopreservation is widely used in animal breeding, the technique is still suboptimal. The population of spermatozoa surviving the procedure experiences changes attributed to alteration in their redox regulation. In order to expand our knowledge regarding this particular aspect, the proteome in fresh and frozen thawed aliquots of equine spermatozoa was studied to identify the proteins most severely affected by the procedure. If alteration of redox regulation is a major factor explaining cryodamage, proteins participating in redox regulation should be principally affected. Using a spli...
Overbeck W, Jäger K, Schoon HA, Witte TS.Endometritis constitutes a major problem in managing broodmares. The histological occurrence of polymorphonuclear neutrophils (PMNs) in the stratum compactum of the endometrium is accepted as the reference standard to diagnose endometritis in mares. The objective of this study was to determine the distribution of PMNs within different sampling locations of the uterus by cytological examinations and to compare it with PMN numbers in endometrial biopsies of the corresponding location. Cytological and endometrial samples were obtained from 37 uteri within 2 ± 1 hours after slaughter through smal...
Minervini F, Lacalandra GM, Filannino A, Garbetta A, Nicassio M, Dell'aquila ME, Visconti A.Fumonisin B1 (FB1) is a mycotoxin produced by Fusarium species that exerts its toxic effect through interference with the sphingolipid pathway by inhibiting ceramide synthase. A FB1-dependent sperm toxicity was reported in boars. No information on FB1-related reproductive toxicity in stallions, the most sensitive animal species, has been reported. The aim of the present study was to assess the in vitro toxicity of FB1 on fresh and frozen-thawed equine spermatozoa by analyzing sperm viability, chromatin stability (SCSA) and reactive oxygen species (ROS) production by flow cytometry and sperm mo...
Sevinga M, Barkema HW, Stryhn H, Hesselink JW.During the foaling seasons of 1999 and 2000, the incidence of retained placenta in 495 normal parturitions of 436 Friesian brood mares was studied. Retained placenta was defined as a failure to expel all fetal membranes within 3 h of the delivery of the foal. Furthermore, the sex of the foal, month of breeding, sire and dam's sire, age of the mare, and time of day of foaling, were studied as factors that might be associated with retained placenta in Friesian mares after normal foalings, and with gestational length. The analysis was carried out using marginal logistic regression, and mixed line...
Hess MF, Roser JF.The large amount of testicular estrogens produced by the stallion is unique compared to the amounts found in other domestic species. Although the cellular locale of the cytochrome P450 aromatase (P450arom) enzyme that converts C19 androgens to C18 estrogens has been identified in the Leydig cell of adult equine testis, the location in the immature equine testis is not known. The goal of this work was to localize the enzyme in colts and stallions during sexual development. Testes were obtained from prepubertal (n=7), pubertal (n=6), and postpubertal (n=8) colts and stallions during both the bre...
Almeida J, Conley AJ, Mathewson L, Ball BA.In the mammalian testis, Leydig cells are primarily responsible for steroidogenesis. In adult stallions, the major endocrine products of Leydig cells include testosterone and estrogens. 3β-hydroxysteroid dehydrogenase/Δ(5)-Δ(4)-isomerase (3βHSD) and 17α-hydroxylase/17,20-lyase (P450c17) are two key steroidogenic enzymes that regulate testosterone synthesis. Androgens produced by P450c17 serve as substrate for estrogen synthesis. The aim of this study was to investigate localization of the steroidogenic enzymes P450c17, 3βHSD, and P450arom and to determine changes in expression during dev...
Pickett BW, Sullivan JJ, Byers WW, Pace MM, Remmenga EE.The effect of centrifugation of diluted and undiluted semen on equine and bovine spermatozoan motility and fertility was examined, as was the effect of seminal plasma and dilution on stallion spermatozoa during incubation before and after freezing. Centrifugation at 370 g or 829 g was not detrimental (P greater than 0.05) to prefreeze or postfreeze motility if a final concentration of 10% seminal plasma was present. A reduction of seminal plasma from 10% to 2% significantly (P smaller than 0.05) reduced motility. A centrifugal force of 956 g significantly reduced prefreeze but not postfreeze m...
Dell'Aquila ME, Albrizio M, Maritato F, Minoia P, Hinrichs K.Follicle atresia and granulosa cell apoptosis may be related to oocyte meiotic and developmental competence. We analyzed the relationships among granulosa cell apoptosis, initial cumulus morphology, oocyte nuclear maturation in vitro, and pronucleus formation after intracytoplasmic sperm injection (ICSI) in the horse. For each follicle, the size was measured and granulosa cells were used for DNA laddering analysis. Oocytes were evaluated for cumulus morphology, cultured for in vitro maturation, and submitted to ICSI. Apoptosis was categorized as absent, intermediate, or advanced according to t...
Maxwell WM, Evans G, Hollinshead FK, Bathgate R, De Graaf SP, Eriksson BM, Gillan L, Morton KM, O'Brien JK.Sex-sorting of mammalian spermatozoa has applications for genetic improvement of farm animals, in humans for the control of sex-linked disease, and in wildlife as a captive management strategy and for the re-population of endangered species. Considerable research has been undertaken worldwide on the Beltsville sperm sexing technology, the only effective method for pre-selection of sex of offspring. The combination of this method with assisted reproductive technologies has resulted in the birth of offspring in a wide range of animals, including cattle, the only livestock species in which sperm ...
Varrasso A, Dynon K, Ficorilli N, Hartley CA, Studdert MJ, Drummer HE.To develop and validate specific, sensitive and rapid (< 8 hour) diagnostic tests using polymerase chain reaction (PCR) for the diagnosis of abortion and respiratory disease caused by equine herpesvirus 1 (EHV1; equine abortion virus) and EHV4 (equine rhinopneumonitis virus). Methods: Primer sets based on nucleotide sequences encoding glycoprotein H (gH) of EHV1 and gB of EHV4 were designed and used in single round and second round (seminested) PCRs, and in a multiplex PCR for the diagnosis of EHV1 and EHV4 infections. Methods: Oligonucleotide primers were designed for each virus, PCR condi...
Galvão A, Valente L, Skarzynski DJ, Szóstek A, Piotrowska-Tomala K, Rebordão MR, Mateus L, Ferreira-Dias G.Regulation of immune-endocrine interactions in the equine endometrium is not fully understood. The aims of the present study were to: (1) investigate the presence of tumour necrosis factor alpha (TNF), interferon gamma (IFNG), Fas ligand (FASLG) and their receptors in the mare endometrium throughout the oestrous cycle; and (2) assess endometrial secretory function (prostaglandins), angiogenic activity and cell viability in response to TNF, oestradiol (E2), progesterone (P4) and oxytocin (OXT). Transcription of TNF and FASLG mRNA increased during the early and late luteal phase (LP), whereas IF...
Papas M, Govaere J, Peere S, Gerits I, Van de Velde M, Angel-Velez D, De Coster T, Van Soom A, Smits K.Anti-Müllerian hormone (AMH) reflects the population of growing follicles and has been related to mammalian fertility. In the horse, clinical application of ovum pick-up and intracytoplasmic sperm injection (OPU-ICSI) is increasing, but results depend largely on the individuality of the mare. The aim of this study was to assess AMH as a predictor for the OPU-ICSI outcome in horses. Therefore, 103 mares with a total follicle count above 10 were included in a commercial OPU-ICSI session and serum AMH was determined using ELISA. Overall, the AMH level was significantly correlated with the number...
Jasiński T, Zdrojkowski Ł, Kautz E, Juszczuk-Kubiak E, Ferreira-Dias G, Domino M.Endometrosis is an important mares' disease which considerably decreases their fertility. As classic endometrial classification methods might be insufficient for tissue pathological evaluation, further categorization into active/inactive and destructive/non-destructive types was developed by Hoffmann and others. This study aimed to compare NF-κB pathway genes transcription among histopathological types of endometrosis, following Hoffmann and co-authors' classification. Endometrial samples, collected postmortem from cyclic mares ( = 100) in estrus or diestrus, were classified histologically an...
Macías García B, González Fernández L, Ortega Ferrusola C, Morillo Rodríguez A, Gallardo Bolaños JM, Rodríguez Martinez H, Tapia JA....Fatty acids and plasmalogens were extracted from the phospholipids of the plasma membrane of stallion spermatozoa, to determine their relation with sperm quality after freezing and thawing. Sperm quality was rated using a quality index that combined the results of the analysis of sperm motility and velocity (CASA analysis), membrane status and mitochondrial membrane potential (flow cytometry) post thaw. Receiving operating system (ROC) curves were used to evaluate the value of specific lipid components of the sperm membrane herein studied as forecast of potential freezeability. From all parame...
Schramme AR, Pinto CR, Davis J, Whisnant CS, Whitacre MD.Current therapy protocols to treat persistent post mating endometritis and retained fetal membranes in mares typically include the administration of ecbolic drugs. Evaluation of the pharmacokinetics and tolerability of carbetocin, a long-acting oxytocin analogue, after i.v. administration is required. Objective: To determine the pharmacokinetic parameters (principally half-life) of carbetocin in horses. Methods: Five mature mares and one gelding received 0.175 mg carbetocin i.v. All animals were monitored periodically throughout the study for elevation in rectal temperature, heart rate, respir...
Gonzalez-Castro RA, Carnevale EM.We determined if microfluidic sorting (MF) of frozen-thawed stallion sperm improves sperm population characteristics and results in embryo development after intracytoplasmic sperm injection (ICSI). The efficiency and efficacy of MF sperm separation was evaluated by comparing pre- and post-separation sperm population variables. Procedural comparisons were performed after sorting with MF, single-layer colloidal centrifugation (SLC) or swim-up (SU), and cleavage and embryo development were evaluated after ICSI using MF-sorted sperm. In Experiment 1, when compared to the original sperm sample, MF ...
Hendriks WK, Colleoni S, Galli C, Paris DBBP, Colenbrander B, Stout TAE.Intracytoplasmic sperm injection is the technique of choice for equine IVF and, in a research setting, 18-36% of injected oocytes develop to blastocysts. However, blastocyst development in clinical programs is lower, presumably due to a combination of variable oocyte quality (e.g. from old mares), suboptimal culture conditions and marginal fertility of some stallions. Furthermore, mitochondrial constitution appears to be critical to developmental competence, and both maternal aging and invitro embryo production (IVEP) negatively affect mitochondrial number and function in murine and bovine emb...
