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Theriogenology.

Periodical
Reproductive Medicine
Veterinary Medicine
Animal Population Groups
Animals
Pregnancy Complications
Reproduction
Publisher:
Geron-X.. [New York, N.Y.?] : Elsevier
Frequency: Sixteen no. a year
Country: United States
Language: English
Start Year:1974 -
ISSN:
0093-691X (Print)
1879-3231 (Electronic)
0093-691X (Linking)
Impact Factor
2.8
2022
NLM ID:0421510
(OCoLC):02504829
(DNLM):T08780000(s)
Coden:THGNBO
Classification:W1 TH677K
Harvesting, processing, and evaluation of in vitro-manipulated equine preantral follicles: A review.
Theriogenology    July 2, 2020   Volume 156 283-295 doi: 10.1016/j.theriogenology.2020.06.044
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...
Embryo technologies in donkeys (Equus Asinus).
Theriogenology    July 1, 2020   Volume 156 130-137 doi: 10.1016/j.theriogenology.2020.06.041
Panzani D, Fanelli D, Camillo F, Rota A.In industrialized countries, the donkey population had a dramatic decrease during the last century and this has brought almost all European donkey breeds to risk. Embryo technologies have already been employed as a tool for the conservation of endangered equid species (e.g. Equus Przewalskii). Today it is possible to obtain pregnancies after the transfer of donkey embryos in synchronized recipients, even though embryo transfer is not widely used as a reproductive technique in this species. So far, very few foals are born after transfer of cryopreserved embryos. To date, no pregnancies have bee...
Supportive techniques to investigate in vitro culture and cryopreservation efficiencies of equine ovarian tissue: A review.
Theriogenology    July 1, 2020   Volume 156 296-309 doi: 10.1016/j.theriogenology.2020.06.043
Aguiar FLN, Gastal GDA, Alves KA, Alves BG, Figueiredo JR, Gastal EL.During the reproductive lifespan of a female, only a limited quantity of oocytes are naturally ovulated; therefore, the mammalian ovary possesses a substantial population of preantral follicles available to be handled and explored in vitro. Hence, the manipulation of preantral follicles enclosed in ovarian tissue aims to recover a considerable population of oocytes of high-value animals for potential application in profitable assisted reproductive technologies (ARTs). For this purpose, the technique of preantral follicle in vitro culture (IVC) has been the most common research tool, achievin...
Effect of Mycobacterium cell wall fraction on endometrial histomorphometry of mares resistant and susceptible to persistent breeding-induced endometritis.
Theriogenology    June 30, 2020   Volume 156 2-10 doi: 10.1016/j.theriogenology.2020.06.034
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...
Equine seminal plasma and sperm membrane: Functional proteomic assessment.
Theriogenology    June 14, 2020   Volume 156 70-81 doi: 10.1016/j.theriogenology.2020.06.014
Guasti PN, Souza FF, Scott C, Papa PM, Camargo LS, Schmith RA, Monteiro GA, Hartwig FP, Papa FO.During ejaculation, a large amount of seminal plasma proteins interact with the sperm membrane, leading to a series of biochemical and structural changes implicated in sperm function and gamete interaction. However, the roles of the majority of these proteins remain unknown. This study aimed to investigate the proteome and functionality of the major equine proteins of seminal plasma and the sperm membrane. Seminal plasma and enriched-membrane proteins (150 μg) were separated by two-dimensional gel electrophoresis, and the respective maps were analyzed. Protein identification was performed by...
Assessment of the anti-inflammatory and engraftment potential of horse endometrial and adipose mesenchymal stem cells in an in vivo model of post breeding induced endometritis.
Theriogenology    June 14, 2020   Volume 155 33-42 doi: 10.1016/j.theriogenology.2020.06.010
Navarrete F, Saravia F, Cisterna G, Rojas F, Silva PP, Rodríguez-Alvarez L, Rojas D, Cabezas J, Mançanares ACF, Castro FO.Horse mesenchymal stem cells (MSC) are potential anti-inflammatory tools for post-breeding induced endometritis (PBIE). In this research MSCs isolated from the endometrium or subcutaneous fat of the same donors were infused iu into mares with PBIE for assessment of their anti-inflammatory action and engraftment. PBIE was induced in nine gynecologically healthy mares by iu infusion of 500 million dead sperm in saline. Inflammatory markers were analyzed in uterine lavages and biopsies immediately before (phase I) and 3 h after infusion of sperm (phase II). Measurements: polymorph nuclear cells ...
Equine endometrial development during late fetal and postnatal periods.
Theriogenology    June 10, 2020   Volume 156 155-161 doi: 10.1016/j.theriogenology.2020.06.006
Fraser NS, Wilborn RR, Johnson AK, Braden TD, Wiley AA, Canisso IF, Bartol FF.Equine uterine development, including endometrial histogenesis, begins prenatally and is completed postnatally. Little is known about this process in the horse. Uterine tissue was acquired from 38 foals, ranging in developmental age from gestational day (GD) 300 to postnatal day (PND) 180, for assessment of endometrial histogenesis. Patterns of endometrial cell proliferation were evaluated by multispectral imaging of uterine tissue sections stained immunofluorescently for Ki-67. Labeling index (LI, % labeled cells) for Ki-67 was calculated for each endometrial cell compartment (luminal epithel...
Heat SHOCK proteins in equine spermatozoa: Expression and correlation to kinetic and environmental parameters.
Theriogenology    June 6, 2020   Volume 155 185-196 doi: 10.1016/j.theriogenology.2020.05.042
Albrizio M, Lacalandra GM, Volpe S, Nicassio M, Cinone M.Heat Shock Proteins are chaperones primary involved in the repair of cellular damages induced by temperature. The harmful effect of temperature on the male gonad is well known, on the contrary knowledge on the effects of the environment on semen quality are still insufficient. The aim of this paper was to learn more about the role of HSPs and the environment in modulating the physiology of equine male gonads. We showed a detailed analysis of equine semen characteristic and the expression level of three HSPs (60-70-90) over a one-year period analyzing the effects of temperature and humidity and...
Serum concentrations and testicular expressions of insulin-like peptide 3 and Anti-Müllerian hormone in normal and cryptorchid male horses.
Theriogenology    May 23, 2020   Volume 154 135-142 doi: 10.1016/j.theriogenology.2020.05.026
Tsogtgerel M, Komyo N, Murase H, Hannan MA, Watanabe K, Ohtaki T, Tsumagari S, Kawate N, Nambo Y.Insulin-like peptide 3 (INSL3) is an important hormone for testicular descent during embryonic development and a factor for assessing functional status of Leydig cells of testes, but there is limited number of equine studies. Anti-Müllerian hormone (AMH) is a useful diagnostic marker for cryptorchidism in horses. This study aimed to compare serum concentrations and testicular expression intensity of INSL3 and AMH in intact and cryptorchid male horses. Serum INSL3 concentrations in intact (n = 9; mean ± SEM, 19.9 ± 5.9 ng/mL) and noncastrated unilateral cryptorchid (UC) male horses (n...
The advantages of using cytospins of uterine lavage fluid for the diagnosis of equine endometritis.
Theriogenology    May 21, 2020   Volume 154 143-151 doi: 10.1016/j.theriogenology.2020.05.008
Marcos R, Ribeiro T, Santos M, Correia-Gomes C, Macedo S, Guimarães T, Blielbernicht M, Caniatti M, Rocha A.Uterine lavage (UL) is a routine diagnostic procedure for endometritis. In UL the fluid is centrifuged and the sediment smeared. Samples prepared in cytocentrifuges, the so-called "cytospins", are useful for evaluating cells in fluids, but never been used in UL. The aim of this study was to assess the usefulness of cytospins after UL, comparing automatic versus manual cytocentrifuges, and to determine their value for the diagnosis of endometritis. The study was divided in two parts. Firstly, UL was performed in 16 mares and a small part of the retrieved fluid was cytocentrifuged in an automati...
Erratum to “Power Doppler can detect the presence of 7-8 day conceptuses prior to flushing in an equine embryo transfer program” [Theriogenology 145C (15 March 2020) 1-9].
Theriogenology    May 19, 2020   Volume 151 159-161 doi: 10.1016/j.theriogenology.2020.04.012
Nieto-Olmedo P, Martín-Cano FE, Gaitskell-Phillips G, Ortiz-Rodríguez JM, Peña FJ, Ortega-Ferrusola C.No abstract available
The influences of cycle stage and pregnancy upon cell glycosylation in the endometrium of the mare.
Theriogenology    May 19, 2020   Volume 154 92-99 doi: 10.1016/j.theriogenology.2020.05.007
Jones CJP, Aplin JD, Allen WRT, Wilsher S.From Day 6.5-7 post-conception until its loss around Day 22, the equine embryo is enclosed in a mucinous capsule that prevents direct intercellular interaction between the trophectoderm and uterine epithelium. The embryo is, however, bathed in glycoprotein-rich secretions. In this study, lectin histochemistry was used to characterise the distribution and glycan composition of uterine glycoproteins destined for secretion, and to ascertain the local effect of an embryo on glycosylation in the endometrium. Endometrial biopsies were taken from mares in estrus, on Days 5, 8, 12 and 15 of diestrus, ...
Prostaglandins effect on matrix metallopeptidases and collagen in mare endometrial fibroblasts.
Theriogenology    May 11, 2020   Volume 153 74-84 doi: 10.1016/j.theriogenology.2020.04.040
Szóstek-Mioduchowska AZ, Baclawska A, Rebordão MR, Ferreira-Dias G, Skarzynski DJ.An increasing number of studies have shown that prostaglandins (PGs) exert multiple regulatory actions in the processes associated to tissue remodeling and fibrosis. Extracellular matrix (ECM) turnover is mediated by matrix metallopeptidases (MMPs). The knowledge about the regulation of their expression in mare endometrium is still limited. Thus, the aim of this study was to investigate whether: (i) profibrotic transforming growth factor (TGF)-β1 modulates PG production in equine endometrium; and (ii) PGE and PGF modulate MMPs, their tissue inhibitors (TIMPs), and collagen 1 (COL1) expression...
A practical guide to estimate the age of the early CL by ultrasonography in mares examined for the first time to be used as recipients in a commercial embryo transfer program.
Theriogenology    May 6, 2020   Volume 153 48-53 doi: 10.1016/j.theriogenology.2020.05.003
Cuervo-Arango J, Martín-Peláez MS, Claes AN.The echogenicity of the early CL undergoes obvious changes during the first 5 days of luteal development. This phenomenon could potentially be used to estimate the age of the CL in mares which have not been checked frequently for the diagnosis of ovulation. The objective of this retrospective study was to determine the accuracy of estimating the age of the early CL (Day 0 to Day 4; Day 0 = Day of ovulation) in recipient mares examined for the first time in the breeding season to be used in a commercial embryo transfer (ET) program. The post-transfer pregnancy rate and embryo loss of 28 recip...
Luteinizing hormone and ovarian steroids affect in vitro prostaglandin production in the equine myometrium and endometrium.
Theriogenology    May 6, 2020   Volume 153 1-8 doi: 10.1016/j.theriogenology.2020.04.039
Piotrowska-Tomala KK, Jonczyk AW, Skarzynski DJ, Szóstek-Mioduchowska AZ.Prostaglandins (PGs) play crucial roles in the regulation of the oestrus cycle and establishment of pregnancy in animals. Luteinizing hormone (LH) and ovarian steroids are involved in regulating endometrial PG production in many species. Their effects on PG production and associated pathways in the mare myometrium and endometrium are the subjects of our interest. This study aimed to evaluate the specific effects of LH and ovarian steroids on equine myometrial and endometrial tissues on (i) PGE and PGF secretion and (ii) transcription of genes encoding specific enzymes responsible for PG synthe...
Kisspeptin/GnRH1 system in Leydig cells of horse (Equus caballus): Presence and function.
Theriogenology    April 15, 2020   Volume 152 1-7 doi: 10.1016/j.theriogenology.2020.04.006
Petrucci L, Maranesi M, Verini Supplizi A, Dall'Aglio C, Mandara MT, Quassinti L, Bramucci M, Miano A, Gobbetti A, Catone G, Boiti C, Zerani M.The objectives of this study were to evaluate in horse testes the expression of kisspeptin (KiSS) and GnRH1 neuropeptides and their cognate receptors, KiSS1R and GnRH1R, as well as their action on testosterone, GnRH1, prostaglandin F2α (PGF2α), and PGE2 synthesis and cyclooxygenase 1 (COX1) and COX2 activity by Leydig cells in vitro. Testes were obtained from 9 sexually mature horses by surgical castration. Immunohistochemistry, evidenced the presence of KiSS, KiSS1R, GnRH, and GnRH1R in Leydig cells, whereas germinal and Sertoli cells were positive only for GnRH1. Transcripts for both neur...
Harem stallion changes are not associated with diminished reproductive performance of females in semi-feral Konik polski horses (Equus caballus).
Theriogenology    March 26, 2020   Volume 151 1-6 doi: 10.1016/j.theriogenology.2020.03.033
Jaworska J, Jaworski Z, McDonnell SM, Górecka-Bruzda A.Infanticide, related to a stallion's aggression toward a foal sired by another stallion, and feticide related to a new stallion's aggression and/or pheromonal influence (the Bruce effect) inducing loss of a fetus sired by another stallion, a female's counteraction to infanticide, have been proposed for domestic horses (Equus caballus) in human-managed conditions. The aim of the present study was, in conditions close to natural, to investigate the influence of the natural succession of a harem stallion on the mares' subsequent reproductive performance. In a population of semi-feral Konik polski...
Effect of warming method on embryo quality in a simplified equine embryo vitrification system.
Theriogenology    March 19, 2020   Volume 151 151-158 doi: 10.1016/j.theriogenology.2020.03.012
Canesin HS, Ortiz I, Rocha Filho AN, Salgado RM, Brom-de-Luna JG, Hinrichs K.Equine embryo vitrification is still not a well-established technique in equine practice. Notably, little work has been done on the effect of the warming system on viability of vitrified embryos. Our goal was to evaluate the effect of warming without cryoprotectants on in vitro - produced (IVP) embryo viability in culture, quality assessment parameters, and pregnancy after transfer. Equine IVP blastocysts were vitrified using commercial embryo vitrification media and a semi-closed vitrification device. In Exp. 1, we evaluated two warming temperatures (room temperature, RT, ∼22 °C; and 38Â...
Immunological memory and tolerance at the maternal-fetal interface: Implications for reproductive management of mares.
Theriogenology    February 28, 2020   Volume 150 432-436 doi: 10.1016/j.theriogenology.2020.02.043
Antczak DF.The development of placentation that coincided with the evolution of mammals presented new challenges to the transmission of life from one generation to the next, particularly with regard to the possibility of maternal immunological recognition and destruction of the developing conceptus. The balance between immunity and tolerance dominates the immunological relationship between mother and fetus during mammalian pregnancy, and the focal point of this relationship lies at the interface between the trophoblast cells that comprise the outermost layer of the placenta and the maternal endometrial t...
Seasonal variations in sperm DNA fragmentation and pregnancy rates obtained after artificial insemination with cooled-stored stallion sperm throughout the breeding season (spring and summer).
Theriogenology    February 21, 2020   Volume 148 89-94 doi: 10.1016/j.theriogenology.2020.02.032
Crespo F, Quiñones-Pérez C, Ortiz I, Diaz-Jimenez M, Consuegra C, Pereira B, Dorado J, Hidalgo M.The aim of this study was to assess seasonal variations during different periods of the breeding season (spring and summer) on stallion sperm DNA fragmentation and in vivo fertility associated with cooled-stored semen samples. Ejaculates were collected from eleven stallions and assessed for sperm motility (assessed by computer-assisted sperm analysis) and plasma membrane integrity (evaluated under fluorescence microscopy). Sperm DNA fragmentation (evaluated by the Sperm Chromatin Dispersion test) was assessed in cooled-stored semen at 5 °C for up to 24 h. Artificial insemination was perfor...
Prediction of the onset of parturition in horses and cattle.
Theriogenology    February 19, 2020   Volume 150 308-312 doi: 10.1016/j.theriogenology.2020.01.072
Nagel C, Aurich J, Aurich C.Economic losses due to dystocia or neonatal death as well as animal welfare and ethical concerns demand a reliable prediction of parturition with the aim to improve survival of the dam and her offspring. In this review, estimation of gestational age by ultrasound and prediction of parturition based on clinical signs, behaviour changes and changes in body temperature, composition of mammary gland secretions as well as hormonal changes are discussed in comparison between horses and cattle. Based on the physiological changes associated with the end of gestation and onset of labor, several systems...
Impact of the corpus luteum on survival of the developing embryo and early pregnancy in mares.
Theriogenology    February 19, 2020   Volume 150 374-381 doi: 10.1016/j.theriogenology.2020.02.011
Pinto CRF.It has been more than a hundred years that studies aiming to elucidate the processes involved in cyclicity and pregnancy pointed out the requirement of ovaries and corpora lutea for embryo survival and pregnancy establishment. For horses, luteal progesterone is essential for pregnancy only during the first trimester. This progestational support is complex among domestic animals as ovarian luteal function is further enhanced by the LH-action role of equine chorionic gonadotropin (eCG) starting ∼ on Day 35 of pregnancy. Increased eCG secretion leads to the formation of supplementary corpora ...
Effects of glucose concentration in semen extender and storage temperature on stallion sperm quality following long-term cooled storage.
Theriogenology    February 11, 2020   Volume 147 1-9 doi: 10.1016/j.theriogenology.2020.02.007
Hernández-Avilés C, Love CC, Serafini R, Ramírez-Agámez L, Friedrich M, Ghosh S, Teague SR, LaCaze KA, Brinsko SP, Varner DD.In Experiment 1, the effects of glucose concentration in extender (0 mM, 67 mM, 147 mM, 270 mM; G0, G67, G147, and G270, respectively) and storage temperature of extended semen (5, 10, 15 and 20 °C) were evaluated after storage for up to 5 days (T0h to T120h). For all time points tested, mean total (TMOT) and progressive (PMOT) sperm motility were lower in G0 than all other treatment groups (P < 0.05). Mean curvilinear velocity (VCL) was lower in G0 than other treatment groups at all time points tested except T0h (P  0.05). Mean TMOT and PMOT, were lower for semen stored at 20 °...
X chromosome aneuploidy and micronuclei in fertile mares.
Theriogenology    February 10, 2020   Volume 147 34-38 doi: 10.1016/j.theriogenology.2020.02.020
Pieńkowska-Schelling A, Kaul A, Schelling C.Abnormalities of chromosomes are an important and well documented cause of disorders of sexual development, fertility problems and congenital anomalies in mammals. Detection of low-level 63,X/64,XX mosaicism during routine cytogenetic evaluation is a challenge because its clinical significance is not yet fully clear. This study describes the prevalence and levels of 63,X mosaicism for a cohort of fertile mares and compares the results with eight problem mares for which no clinical cause of sub-fertility was found. The study design allowed for the analysis of micronuclei which are biomarkers of...
Biomarkers for placental disease in mares.
Theriogenology    January 30, 2020   Volume 150 302-307 doi: 10.1016/j.theriogenology.2020.01.073
Canisso IF, Loux SC, Lima FS.Placentitis is an important cause of abortion, stillbirth, and neonatal death in horses. The diagnosis of placentitis is based on occurrence of clinical signs (premature mammary gland development and vulvar discharge) and ultrasonography of the caudal placental pole. However, early and subtle cases can be missed. In the last few years, several studies have provided objective means of diagnosing placentitis in mares with single or serial measurements of blood markers. Among the markers evaluated the steroids produced by the fetoplacental unit have been shown to change in association with placen...
Expression of genes involved in the NF-κB-dependent pathway of the fibrosis in the mare endometrium.
Theriogenology    January 29, 2020   Volume 147 18-24 doi: 10.1016/j.theriogenology.2020.01.055
Domino M, Jasinski T, Kautz E, Juszczuk-Kubiak E, Ferreira-Dias G, Zabielski R, Sady M, Gajewski Z.Equine endometrosis is a multifactorial chronic degenerative condition, considered to be one of a major causes of equine infertility. The formation of periglandular fibrosis seems to be linked to chronic inflammation of the mare endometrium in a paracrine way and in a response to numerous forms of inflammatory stimuli elicit the net deposition of extracellular matrix (ECM) around the endometrial glands and stroma. We hypothesized some of these stimuli, such as monocyte chemoattractant protein-1 (MCP-1), interleukin-6 (IL-6), and hyaluronan synthases (HASs), may share the nuclear factor-κB (NF...
Neutrophils, monocytes and other immune components in the equine endometrium: Friends or foes?
Theriogenology    January 13, 2020   Volume 150 150-157 doi: 10.1016/j.theriogenology.2020.01.018
Skarzynski DJ, Szóstek-Mioduchowska AZ, Rebordão MR, Jalali BM, Piotrowska-Tomala KK, Leciejewska N, Łazarczyk M, Ferreira-Dias GM.The innate and adaptive immune mechanisms are key components of regulation of reproductive physiological function and uterine disorders in equine uterus. The predominant immunological response in equine endometrium, characterized by an innate immune response, occurs under estrogens influence, in the follicular phase. Although, the increase in immune-related genes in equine endometrium during estrus has been suggested to play a role in uterine clearance after mating, immune cells and their product, i.e. cytokines play also mandatory role in the luteal development and maintenance, regression of ...
Power Doppler can detect the presence of 7-8 day conceptuses prior to flushing in an equine embryo transfer program.
Theriogenology    January 9, 2020   Volume 145 1-9 doi: 10.1016/j.theriogenology.2020.01.015
Nieto-Olmedo P, Martín-Cano FE, Gaitskell-Phillips G, Ortiz-Rodríguez JM, Peña FJ, Ortega-Ferrusola C.In order to determine whether differences in uterine blood flow between pregnant and non-pregnant mares can be used to predict the presence of the equine embryo prior to flushing in an embryo transfer program, power Doppler ultrasonography was used on a total of 52 mares on days 7 or 8 post-ovulation. Computer analysis of Doppler images was subsequently performed using ImageJ v1.48 software. Vascular perfusion of the endometrium was analyzed using spot meter techniques, measuring mean pixel intensity and area of blood flow. Mares with positive flushings presented a higher uterine blood flow ar...
Does semen quality change after local treatment of seminal vesiculitis in stallions?
Theriogenology    December 27, 2019   Volume 144 139-145 doi: 10.1016/j.theriogenology.2019.12.010
Sancler-Silva YFR, Monteiro GA, Ramires-Neto C, Freitas-Dell'aqua CP, Crespilho AM, Franco MMJ, Silva-Junior ER, Cavalero TMS, Scheeren VFC, Papa FO.Inflammation of the seminal vesicle interferes with fertility and is a persistent problem that is difficult to treat. The aim of this study was to evaluate the semen quality of 5 stallions with seminal vesiculitis before and after local treatment. All stallions were endoscopically treated for seminal vesiculitis during 10 consecutive days. The glandular lumen was accessed and flushed with a Ringer Lactate solution prior to antibiotic infusion. The antibiotic was selected based on the antibiogram from bacterial culture of samples previously collected from the seminal vesicles. The kinetic param...
Seasonal changes in ROS concentrations and sperm quality in unfrozen and frozen-thawed stallion semen.
Theriogenology    December 24, 2019   Volume 144 89-97 doi: 10.1016/j.theriogenology.2019.12.016
Mislei B, Bucci D, Malama E, Bollwein H, Mari G.Oxidative stress is regarded as an important cause of sperm damage during cryopreservation. However, seasonal changes in oxidative status in unfrozen and frozen-thawed stallion sperm have not been well established. We tested the hypothesis that sperm ROS concentrations and lipid peroxidation change between breeding and non-breeding seasons and influence quality of unfrozen and frozen-thawed sperm. Eighteen ejaculates from six Warmblood stallions (8-21 y) known to be fertile, were collected in winter and summer and processed for freezing. After 90 min at +4 °C, some straws from each ejacu...
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