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Topic:Ovulation

Ovulation in horses is a physiological process involving the release of an ovum from the ovary, typically occurring during the mare's estrous cycle. This process is regulated by hormonal changes, primarily involving luteinizing hormone (LH) and follicle-stimulating hormone (FSH). Ovulation is a key component of equine reproduction, influencing breeding management and fertility outcomes. The timing and detection of ovulation are critical for optimizing breeding strategies, often involving techniques such as ultrasound examination and hormone assays. This page compiles peer-reviewed research studies and scholarly articles that investigate the mechanisms, detection methods, and implications of ovulation in equine reproductive health.
Ovarian and Uterine Dynamics During the Estrous Cycle in Criollo Breed Mares.
Journal of equine veterinary science    September 29, 2022   Volume 118 104131 doi: 10.1016/j.jevs.2022.104131
Duval LH, Rechsteiner SMF, Gastal GDA, Gastal MO, Mattos RC, Gastal EL.After having been brought to America 400 years ago, the Criollo horse reproduced freely for centuries in the southern part of the American continent. Roughness, resistance, and endurance are typical characteristics of this breed of horses. Although the natural selection that occurred over the centuries may have positively influenced the fertility and longevity of this breed, information regarding ovarian function and other reproductive aspects of Criollo breed mares under natural or controlled management conditions is scarce. The objectives of the present study were to (1) characterize the fol...
The Correlation of Endogenous Progesterone Concentration in Diestrus on Early Pregnancy Rate in Thoroughbred Mares.
Journal of equine veterinary science    September 15, 2022   Volume 118 104127 doi: 10.1016/j.jevs.2022.104127
Hollinshead FK, Mehegan MK, Gunn A, Nett T, Bruemmer JE, Hanlon DW.The aim of this project was to test the hypothesis that progesterone concentration 5 days after ovulation did not differ between pregnant and nonpregnant Thoroughbred mares on stud farms located in the Waikato region of New Zealand. A prospective cohort study was performed involving five stud farms in the Waikato region of New Zealand during the 2018 breeding season. A total of 275 mares were enrolled in the study. Mares were served by 34 individual stallions. Blood samples were taken from each mare 5 days after ovulation (D0) and measured for progesterone concentration. Early pregnancy was co...
Effect of day of estrus cycle at time of transvaginal follicle aspiration for oocyte recovery on rates of in vitro maturation and blastocyst production after intracytoplasmic sperm injection.
Journal of the American Veterinary Medical Association    July 28, 2022   Volume 260, Issue 13 1683-1689 doi: 10.2460/javma.22.06.0240
Walbornn SR, Felix M, Schnobrich MR, Bradecamp EA, Scoggin CF, Stefanovski D, Hinrichs K.To determine the effect of stage of estrus cycle (day after ovulation) at the time of transvaginal ultrasound-guided follicle aspiration (TVA) on parameters related to the success of in vitro equine embryo production. 14 healthy mares were used; 11 completed the study and were included for analysis. Mares underwent TVA of all follicles ≥ 5 mm diameter at each of 3 timepoints: 7 days after ovulation, 14 days after ovulation, and S-DSF (subordinate to a dominant stimulated follicle), during estrus at 24 hours after gonadotropin administration. The 3 treatments were assigned to each mare in ran...
Luteal Tissue Area and Immunoreactive Concentration of Progesterone in Plasma of Bred and Non-bred Mares.
Journal of equine veterinary science    July 10, 2022   Volume 118 104075 doi: 10.1016/j.jevs.2022.104075
Colombo I, Podico G, Rudolf-Vegas A, Bauersachs S, Canisso IF.Progesterone is pivotal to maintain pregnancy in the first trimester and low concentration ( .05). Multiple ovulations were associated with greater progesterone concentration and luteal tissue area (P = .0001). There was a moderate positive association between the number of ovulations and luteal tissue area (r = 0.54; P = .0001). The lack of change in the progesterone concentration and luteal tissue area between bred and non-bred mares suggests that horse seminal plasma does not affect luteal function in mares. As all mares had progesterone above 4 ng/mL after 5-days post-ovulation; it i...
Follicular-fluid proteomics during equine follicle development.
Molecular reproduction and development    June 27, 2022   Volume 89, Issue 7 298-311 doi: 10.1002/mrd.23622
Ishak GM, Feugang JM, Pechanova O, Pechan T, Peterson DG, Willard ST, Ryan PL, Gastal EL.The complex composition of the follicular fluid (FF), the intimate proximity to the oocyte, and the continual changes in their composition have a major effect on folliculogenesis and oogenesis. To date, the profiling of FF proteomes during follicle selection, development, and ovulation has not been comprehensively investigated. Therefore, a shotgun proteomics approach and bioinformatics analyses were used to profile the proteomes of equine FF harvested in vivo from follicles at the following development stages: predeviation (18-20 mm), deviation (22-25 mm), postdeviation (26-29 mm), preo...
The Kisspeptin analogue C6 induces ovulation in jennies.
Theriogenology    June 15, 2022   Volume 189 107-112 doi: 10.1016/j.theriogenology.2022.06.014
Fanelli D, Beltramo M, Conte G, Cerretini B, Lomet D, Rota A, Aucagne V, Camillo F, Panzani D.Kisspeptins (KPs) are the most potent stimulating neurotransmitters of GnRH release, and consequently KP administration triggers LH and/or FSH release. In small ruminants, KP or its analogs induced an LH surge followed by ovulation in both cyclic and acyclic animals, while in the mare KP only increased LH plasma levels but failed to induce ovulation. This study in jennies compares the endocrinological effects, ovulatory and pregnancy rates of the KP analog C6 and the GnRH analog buserelin acetate. The ovarian activity of nine Amiata jennies was monitored daily by transrectal ultrasound for thr...
Emergence and selection of the dominant follicle and gonadotropin dynamics in postpartum lactating versus non-postpartum cycling mares.
Reproductive biology    May 10, 2022   Volume 22, Issue 2 100618 doi: 10.1016/j.repbio.2022.100618
Pastorello M, Gastal MO, Godoi DB, Gastal EL.Among female livestock, the mare has the shortest interval from parturition to first ovulation. Due to the scarcity of research on postpartum mares, little progress has been made on the characterization of the resumption of ovarian cyclicity after parturition. This study compared follicular and gonadotropin dynamics during follicle emergence and deviation in postpartum lactating (PP Lactating) versus non-postpartum cycling (N-PP Cycling) mares. On the day of parturition, every PP Lactating mare was paired with a N-PP Cycling mare. Comparisons were made by considering the partum-ovulation inter...
Suppression of reproductive behaviour and gonadal function in female horses-An update.
Reproduction in domestic animals = Zuchthygiene    May 1, 2022   Volume 57 Suppl 4, Issue Suppl 4 4-12 doi: 10.1111/rda.14129
Aurich C, Kaps M.The behaviour of mares is often detrimental to their performance resulting in frequent demand for methods to suppress gonadal function. In addition, prevention of unintended reproduction especially in feral horse populations may require methods for suppression of gonadal function. Surgical ovariectomy is a safe method but not an acceptable approach in feral mares and undesired in mares where future breeding is considered. There are different approaches for artificial prolongation of the luteal phase resulting in transient inhibition of oestrus and ovulation. Among those, treatment with natural...
Success rate in a clinical equine in vitro embryo production program.
Theriogenology    April 24, 2022   Volume 187 215-218 doi: 10.1016/j.theriogenology.2022.04.019
Claes A, Stout TAE.In vitro embryo production (IVEP) via Ovum Pick-Up (OPU) and Intracytoplasmic Sperm Injection (ICSI) has become a popular breeding technique in Warmblood mares because of the high success rate and several practical advantages. IVEP offers a solution for a variety of reproductive issues including, but not limited to, sub-fertility in stallions or mares, poor quality or scarce frozen semen, difficulty in synchronizing donor and recipient mares, and inefficient use of recipient mares. In 515 OPU-ICSI sessions performed in our facility in 2021, a mean of 25.9 antral follicles were aspirated yield...
Effect of short-term artificial light and transvaginal progesterone device on first ovulation in late transitional mares.
Journal of equine science    April 19, 2022   Volume 33, Issue 1 1-6 doi: 10.1294/jes.33.1
Bianchi CP, Bruno S, Videla Dorna I, Rodríguez E, Aba MA.In study I, plasma progesterone concentrations were evaluated in anoestrous mares that received an intravaginal progesterone release device (IPRD) for 10 days. Mares were divided into 3 groups based on the dosage of progesterone (0 g, n=3; 1.38 g, n=5; and 1.9 g, n=5). No statistical differences were found in plasma progesterone concentrations between the two doses tested. In study II, the effects of a protocol based on a short program of artificial light combined with an IPRD containing 1.38 g of progesterone on oestrous behaviour and onset of ovulation were evaluated. IPRDs were inserted int...
Ovulatory response to GnRH agonist during early and late fall in mares.
Theriogenology    March 15, 2022   Volume 185 140-148 doi: 10.1016/j.theriogenology.2022.03.003
Spencer KM, Podico G, Megahed AA, Jones KL, Bittar JHJ, Canisso IF.This study aimed to assess the ovulatory response of deslorelin acetate during the fall and the response to PGF2α 8 d post-ovulation. One hundred estrous cycles from 22 mares kept in 40° latitude were evaluated. Mares were checked by transrectal ultrasonography until a preovulatory follicle was detected and ovulation induced with deslorelin acetate. Ovulation was confirmed by ultrasonography performed at 24, 36 h post-induction and then repeated at 2-h intervals post-induction. Serum progesterone concentrations and luteal tissue area were determined daily to assess CL function. A dose of P...
No increase in pregnancy rate of mares after preovulatory deep uterine horn application of misoprostol.
Theriogenology    March 14, 2022   Volume 184 132-139 doi: 10.1016/j.theriogenology.2022.03.005
Donatsch L, Friker B, Sieme H, Kaeser R, Burger D.A potential source of fertility loss in mares is oviductal dysfunction, potentially caused by masses or debris in the lumen, that may prevent either sperm from reaching the fertilization site or the embryo from reaching the uterus. Recently a novel therapeutic method leading to increased pregnancy results was described by infusing misoprostol, a synthetic prostaglandin E, in the uterus of mares with unexplained fertility problems. In this study, we aimed, after examining the compatibility of misoprostol with semen, to evaluate the pregnancy rate after routine preovulatory deep uterine horn app...
Dominant follicle and gonadotropin dynamics before ovulation in postpartum lactating mares.
Molecular reproduction and development    January 9, 2022   Volume 89, Issue 2 113-124 doi: 10.1002/mrd.23552
Gastal MO, Pastorello M, Godoi DB, Gastal EL.The aim of this study was to compare the dominant follicle (DF) and gonadotropin dynamics for 9 days before ovulation in postpartum lactating (PP Lactating) versus non-postpartum cycling (N-PP Cycling) mares. Every PP Lactating mare on the day of parturition was paired with a N-PP Cycling mare, and the data analyses considered the partum-ovulation interval (POI) and the postpartum interovulatory interval (PPIOI) in PP Lactating mares and two interovulatory intervals in N-PP Cycling mares. The results of the present study revealed several novel and unique aspects of DF development and FSH and L...
Assessment of anti-Müllerian hormone in mares’ transitional period and in relation to fertility in elderly mares.
Theriogenology    November 30, 2021   Volume 179 97-102 doi: 10.1016/j.theriogenology.2021.11.025
Fouché N, Gerber V, Bruckmaier RM, Erni-Wespi B, Zander Y, Vidondo B, Sieme H, Claes A, Kaeser R, Burger D.Anti-Müllerian Hormone (AMH) has been linked to reproductive tract abnormalities in mares and stallions. This study aimed at evaluating AMH as a biomarker for two reproductive conditions in mares. In the first part of this study, plasma AMH was evaluated as an early indicator of the onset of cyclicity in mares in the transitional period from the anovulatory phase during winter anoestrus to the cyclic phase during the breeding season. Ten mares between 8 and 17 years old were included in the experiment which lasted from mid-February until the end of April. Ovarian activity was monitored with u...
Interrelationship between reproductive hormones and acute phase proteins during estrous cycle and pregnancy in Spanish purebred broodmares.
Veterinary and animal science    October 26, 2021   Volume 14 100212 doi: 10.1016/j.vas.2021.100212
Satué K, Calvo A, Muñoz A, Fazio E, Medica P.In some species, female steroid hormones modify the profile of acute phase proteins (APPs) during the estrous cycle and pregnancy, according to the ovulation, embryonic implantation and placental development; however, nowadays there's no experimental evidence for equine species. Objectives of this study were: to compare the serum amyloid A (SAA), haptoglobin (Hp) and C-reactive protein (CRP) concentrations between cyclic and pregnant mares, and to analyze the influence of estradiol-17β (E) during estrous cycle or estrone sulfate (E) during pregnancy, and progesterone (P) on these proteins to ...
Variables Affecting Veterinary Students’ Ability to Accurately Interpret Ovulation in Live Mare Palpation.
Journal of veterinary medical education    September 27, 2021   Volume 49, Issue 5 618-624 doi: 10.3138/jvme-2021-0031
Fanelli D, Tesi M, Rota A, Panzani D, Camillo F.In a veterinary medicine curriculum, students' hands-on practice is essential but is still considered one of the major deficiencies in veterinary schools in Europe. After theoretical and basic practical training, students, under the control of experienced veterinarians (supervisors), monitored the reproductive cycle of embryo recipients by transrectal palpation and ultrasound. To evaluate the skills of students, the question "Has she ovulated?" was posed when a dominant follicle ≥ 35 mm was recorded in the previous day's examination and a score of 1 or 0 was assigned in the case of a correct...
Systemic and Intrafollicular Androgen Concentrations in Cycling Mares.
Journal of equine veterinary science    September 11, 2021   Volume 106 103759 doi: 10.1016/j.jevs.2021.103759
Satué K, Fazio E, Cravana C, Medica P.The evidence that androgens regulate the folliculogenesis supports the hypothesis that intrafollicular testosterone (T), androstenedione (A) and dehydroepiandrosterone (DHEA) could be modified along follicular growth. The objective of this study was to evaluate the changes and related relationships between systemic and intrafollicular T, A and DHEA in post-deviation and impending ovulation follicles. Sixty ovaries were taken after the slaughter of 30 clinically healthy mares. In according to the sizes, the follicles were classified in 3 different categories, as small (20-30 mm), medium (31-40 ...
Spatial distribution of Gasterophilus pecorum (Diptera) eggs in the desert steppe of the Kalamaili Nature Reserve (Xinjiang, China).
BMC ecology and evolution    September 6, 2021   Volume 21, Issue 1 169 doi: 10.1186/s12862-021-01897-4
Huang H, Zhang K, Shao C, Wang C, Ente M, Wang Z, Zhang D, Li K.The dominant Gasterophilus species in the desert steppe (Xinjiang, China) Gasterophilus pecorum poses a serious threat to the reintroduced Przewalski's horses. We investigated the distribution pattern of G. pecorum eggs in June 2017. Two sampling methods, transect and grid, were used, and the results were analyzed via geostatistics by semivariance. The nest quadrat was used to determine the optimal quadrat size. Eggs were found in 99 quadrats (63.1%) and 187 clusters (1.5%) of Stipa caucasica on the steppe. The mean oviposition count of a cluster was 3.8 ± 1.6. Three-eggs is the mode of w...
Reproductive patterns and follicular waves in postpartum lactating versus non-postpartum cycling mares.
Journal of equine veterinary science    August 13, 2021   Volume 107 103732 doi: 10.1016/j.jevs.2021.103732
Gastal EL, Pastorello M, Godoi DB, Gastal MO.This comparative study between postpartum lactating (PP Lactating) and non-postpartum cycling (N-PP Cycling) mares aimed to characterize reproductive patterns, types and frequencies of follicular waves, corpus luteum and endometrial echotexture dynamics, and the influence of season and body condition. Mares from each group were paired considering the day of parturition of a PP Lactating mare. The partum-ovulation interval (POI) and the postpartum interovulatory interval (PPIOI) were evaluated for PP Lactating mares, and 2 IOIs were evaluated for N-PP Cycling mares. The following observations w...
Assessment of Age Effects on Ovarian Hemodynamics Using Doppler Ultrasound and Progesterone Concentrations in Cycling Spanish Purebred Mares.
Animals : an open access journal from MDPI    August 8, 2021   Volume 11, Issue 8 2339 doi: 10.3390/ani11082339
Requena F, Campos MJAPM, Martínez Marín AL, Camacho R, Giráldez-Pérez RM, Agüera EI.In equine reproduction, accurate and timely detection of the moment of ovulation is of great importance. Power Doppler ultrasound technology is a non-invasive method that enables to assess the morpho-echogenic features and blood flow changes during the estral cycle in mares. The objective of the present study was to evaluate the influence of age on ultrasonographic parameters (follicular diameter, follicular blood flow-FBF, corpus luteum (CL) area and corpus luteum blood flow-CLBF) and blood plasma progesterone concentrations in cycling Spanish Purebred mares (15 less than 8 years old and 15 e...
The Proteome of Equine Oviductal Fluid Varies Before and After Ovulation: A Comparative Study.
Frontiers in veterinary science    August 5, 2021   Volume 8 694247 doi: 10.3389/fvets.2021.694247
Fernández-Hernández P, Marinaro F, Sánchez-Calabuig MJ, García-Marín LJ, Bragado MJ, González-Fernández L, Macías-García B.Equine fertilization cannot be performed in the laboratory as equine spermatozoa do not cross the oocyte's zona pellucida . Hence, a more profound study of equine oviductal fluid (OF) composition at the pre-ovulatory and post-ovulatory stages could help in understanding what components are required to achieve fertilization in horses. Our work aimed to elucidate the proteomic composition of equine OF at both stages. To do this, OF was obtained postmortem from oviducts of slaughtered mares ipsilateral to a pre-ovulatory follicle ( = 4) or a recent ovulation ( = 4); the samples were kept at -80°...
Anti-Müllerian Hormone and OPU-ICSI Outcome in the Mare.
Animals : an open access journal from MDPI    July 5, 2021   Volume 11, Issue 7 doi: 10.3390/ani11072004
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...
The Combination of hCG and GnRH Analog to Hasten Ovulation in Mares Does not Change Luteal Function and Pregnancy Outcome in Embryo Recipient Mares.
Journal of equine veterinary science    June 21, 2021   Volume 105 103691 doi: 10.1016/j.jevs.2021.103691
Segabinazzi LGTM, Oba E, Alvarenga MA.Equine practitioners often prescribe the combined use of hCG and GnRH to hasten ovulation due to presumed synergistic effects. Therefore, this study aimed to test whether the combination of hCG and deslorelin acetate to hasten ovulation in mares would show any effect in inducing ovulation more efficiently than when either drug is used by itself, and to verify whether this association would affect progesterone concentrations; corpus luteum (CL) diameter and blood flow; and pregnancy outcome in recipient mares after embryo transfer (ET). Seventeen mares had the ovulation hastened (≥35 mm folli...
Sperm migration, selection, survival, and fertilizing ability in the mammalian oviduct†.
Biology of reproduction    June 1, 2021   Volume 105, Issue 2 317-331 doi: 10.1093/biolre/ioab105
Mahé C, Zlotkowska AM, Reynaud K, Tsikis G, Mermillod P, Druart X, Schoen J, Saint-Dizier M.In vitro fertilization (IVF) gives rise to embryos in a number of mammalian species and is currently widely used for assisted reproduction in humans and for genetic purposes in cattle. However, the rate of polyspermy is generally higher in vitro than in vivo and IVF remains ineffective in some domestic species like pigs and horses, highlighting the importance of the female reproductive tract for gamete quality and fertilization. In this review, the way the female environment modulates sperm selective migration, survival, and acquisition of fertilizing ability in the oviduct is being considered...
Deslorelin Slow-Release Implants Delay Ovulation and Increase Plasma AMH Concentration and Small Antral Follicles in Haflinger Mares.
Animals : an open access journal from MDPI    May 28, 2021   Volume 11, Issue 6 doi: 10.3390/ani11061600
Kaps M, Okada CTC, Gautier CM, Aurich J, Aurich C.There is an increasing interest in the manipulation of ovarian follicular populations in large domestic animals because this could prove beneficial for assisted reproductive techniques such as ovum pick-up (OPU). The aim of the present study was to evaluate the effects of deslorelin slow-release implants (SRI) on the interovulatory interval, antral follicle count (AFC), number of follicles of different size ranges and plasma anti-Muellerian hormone (AMH) concentration in mares. To synchronize their estrous cycles, Haflinger mares (n = 12) were treated twice with a PGF2α analogue. One day afte...
Switching of follicle destiny so that the second largest follicle becomes dominant in monovulatory species.
Theriogenology    May 20, 2021   Volume 171 147-154 doi: 10.1016/j.theriogenology.2021.05.002
Ginther OJ.During an ovulatory follicular wave in the monovulatory species of heifers, mares, and women, the two largest follicles deviate in diameter at the end of a common follicle growth phase. The largest follicle before deviation becomes the future ovulatory follicle in most ovulatory waves. In 10-30% of the ovulatory waves, the destiny of the two follicles switches just before or at deviation so that the second-largest follicle becomes the future ovulatory follicle, and the largest follicle becomes a subordinate. In FSH-driven switching in heifers, mares, and women, the wave-stimulating FSH surge d...
hCG is more effective than the GnRH agonist buserelin for inducing the first ovulation of the breeding season in mares.
Equine veterinary journal    May 9, 2021   Volume 54, Issue 2 306-311 doi: 10.1111/evj.13455
Fanelli D, Tesi M, Rota A, Beltramo M, Conte G, Giorgi M, Barsotti G, Camillo F, Panzani D.Human Chorionic Gonadotropin (hCG) and Gonadotropin Releasing Hormone agonists (GnRHa) are routinely used to induce ovulation in mares. However, GnRHa efficacy in transitional mares has been suggested to be low. Objective: The aims of this study were as follows: (a) to compare the efficacy of hCG and GnRHa in inducing the first ovulation of the breeding season and (b) to evaluate the correlation between ovulatory response, uterine oedema and teasing score at the time of treatment during the early or late transitional phase. Methods: Randomised controlled superiority trial. Methods: Mares in wi...
Changes Associated with the Peri-Ovulatory Period, Age and Pregnancy in ACTH, Cortisol, Glucose and Insulin Concentrations in Mares.
Animals : an open access journal from MDPI    March 20, 2021   Volume 11, Issue 3 891 doi: 10.3390/ani11030891
Hicks GR, Fraser NS, Bertin FR.Although there are many hormonal changes associated with reproduction, the effects of ovulation and early pregnancy on adrenocorticotropic hormone (ACTH) and insulin concentrations are poorly described. We hypothesise that both ovulation and early pregnancy will alter ACTH and insulin concentrations in healthy mares. Eighteen mares showing no clinical signs suggestive of, or laboratory findings consistent with, pituitary pars intermedia dysfunction PPID and insulin dysregulation (ID) are enrolled. ACTH, cortisol, insulin and glucose concentrations are measured over their peri-ovulatory period,...
Concentrations of progesterone and a PGF2α metabolite during the interovulatory interval compared to the corresponding days of pregnancy in mares.
Theriogenology    February 11, 2021   Volume 165 10-17 doi: 10.1016/j.theriogenology.2021.02.004
Castro T, Jacob JC, Stefani G, Domingues RR, Ginther OJ.The concentrations of progesterone (P4) and a metabolite of PGF2α (PGFM) in mares were compared between the interovulatory interval (IOI; n = 8) and the corresponding days of pregnancy (n = 9). In daily blood samples, P4 increased between the day of ovulation (Day 0) and ∼Day 6 and then gradually decreased until the beginning of luteolysis in the IOI group. Before the beginning of luteolysis, there were no significant differences in P4 concentrations between the IOI and early pregnancy. In the IOI, PGFM concentration on the day before the beginning of luteolysis began to increase (P <Â...
L-Proline: An Effective Agent for Frozen and Post-thawed Donkey Semen Storage.
Journal of equine veterinary science    February 3, 2021   Volume 101 103393 doi: 10.1016/j.jevs.2021.103393
Li N, Yu J, Yang F, Shao Y, Wu S, Liu B, Li M, Wang T, Li J, Zeng S.The aim of this study was to evaluate the effects of L-proline on the extender quality of frozen and post-thawed jackass semen. Jackass (n = 6) semen samples were collected and cryopreserved in gradient concentrations (0-80 mM) of L-proline in extenders; post-thawed semen samples were cultured in L-proline medium for 10 hours at 37°C. For cryopreservation experiment I, the motile parameters, mitochondrial membrane potential (MMP), and plasma membrane, acrosome, and chromatin structure integrities of post-thawed semen were assessed. For culture experiment II, additional ROS contents were ana...