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Topic:Reproduction

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.
Placental alterations in structure and function in intra-uterine growth-retarded horses.
Equine veterinary journal    November 6, 2017   Volume 50, Issue 3 405-414 doi: 10.1111/evj.12761
Robles M, Peugnet PM, Valentino SA, Dubois C, Dahirel M, Aubrière MC, Reigner F, Serteyn D, Wimel L, Couturier-Tarrade A, Chavatte-Palmer P.Following embryo transfer (ET), the size and breed of the recipient mare can affect fetal development and subsequent post natal growth rate and insulin sensitivity in foals. Objective: To investigate placental adaptation in pregnancies where increased or restricted fetal growth was induced through ET between Pony, Saddlebred and Draught horses. Methods: In vivo experiment. Methods: Control Pony (P, n = 21) and Saddlebred (S, n = 28) pregnancies were obtained by artificial insemination. Increased pregnancies were obtained by transferring Pony (P-D, n = 6) and Saddlebred (S-D, n = 8) emb...
Hemodynamics of the corpus luteum in mares during experimentally impaired luteogenesis and partial luteolysis.
Theriogenology    November 4, 2017   Volume 107 78-84 doi: 10.1016/j.theriogenology.2017.10.045
Ferreira JC, Filho LFN, Boakari YL, Canesin HS, Thompson DL, Lima FS, Meira C.The aim of the current project was to characterize the luteal vascularity and the plasma concentrations of progesterone (P4), prolactin (PRL) and 13,14-dihydro-15-keto-PGF (PGFM) in mares with luteal disturbances during early and mid-diestrus. In Experiment 1, twenty-one mares were treated with 2 mL of 0.9% NaCl, or 1 mg Dinoprost, or 10 mg Dinoprost on day two after ovulation (Control-D2, 1/10PGF-D2 and PGF-D2 groups, respectively; n = 7 mares/group). In Experiment 2, similar treatments were performed eight days post-ovulation using a different cohort of 21 mares (Control-D8, 1/10PGF-D8 ...
Half a century of equine reproduction research and application: A veterinary tour de force.
Equine veterinary journal    November 3, 2017   Volume 50, Issue 1 10-21 doi: 10.1111/evj.12762
Allen WR, Wilsher S.Over the past 50 years, per season pregnancy rate in Thoroughbred mares have risen from 70 to >90% and foaling rates from 55 to >80%. Some of the significant pharmacological treatments and diagnostic methods which have driven this marked improvement in breeding efficiency are reviewed. They include the application of artificial lighting to hasten the onset of ovulatory oestrous cyclicity in early Spring, rapid steroid hormone assays to aid in determining the stage and normality of the mare's cycle, prostaglandin analogues, synthetic progestagens and Gonadotrophin-releasing Hormone (GnRH) anal...
Equine Endometrial Explants Undergo Significant Degenerative Changes in Culture.
Anatomical record (Hoboken, N.J. : 2007)    November 2, 2017   Volume 301, Issue 1 148-153 doi: 10.1002/ar.23701
Schwinghamer R, Massolo A, Knight C, Klein C.This study evaluated equine endometrial explants following 12, 24, and 48 hr in culture. Measurement of an indicator of cell death in explant supernatant, light microscopy, and gene expression of biomarkers of endometrial function and cellular stress were used to compare the effect of six different media on explant viability and morphology. Viability of explants was assessed indirectly through measuring LDH activity in the culture supernatant. Regardless of culture medium composition, a significant increase in LDH activity was observed within 12 hr of culture, indicating occurrence of cell dam...
Sex-steroid receptors, prostaglandin E2 receptors, and cyclooxygenase in the equine cervix during estrus, diestrus and pregnancy: Gene expression and cellular localization.
Animal reproduction science    October 31, 2017   Volume 187 141-151 doi: 10.1016/j.anireprosci.2017.10.018
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...
A fast, low-cost and efficient method for the diagnosis of sperm DNA fragmentation in several species.
Reproduction in domestic animals = Zuchthygiene    October 30, 2017   Volume 53, Issue 1 171-175 doi: 10.1111/rda.13087
Rui BR, Angrimani D, Bicudo LC, Losano J, Nichi M, Pereira R.Sperm DNA fragmentation is a condition that interferes directly in the reproductive efficiency. Currently, there are several methods for assessing the sperm DNA integrity, such as Alkaline Comet, TUNEL and Sperm Chromatin Structure Assay. However, many of these techniques are laborious and require high-precision equipment. Thus, the development of new techniques can optimize the evaluation of sperm DNA damage. Therefore, the aim of this study was to standardize the toluidine blue (TB) stain technique for the analysis of DNA fragmentation of dog, cat, bull, stallion and ram spermatozoa. For thi...
Extreme breedingin horses.
The Veterinary record    October 28, 2017   Volume 181, Issue 17 458 doi: 10.1136/vr.j4910
Denny DJB.No abstract available
Steroidogenic enzyme activities in the pre- and post-parturient equine placenta.
Reproduction (Cambridge, England)    October 24, 2017   Volume 155, Issue 1 51-59 doi: 10.1530/REP-17-0472
Legacki EL, Corbin CJ, Ball BA, Scoggin KE, Stanley SD, Conley AJ.Steroidogenic enzymes in placentas shape steroid hormone profiles in the maternal circulation of each mammalian species. These include 3β-hydroxysteroid dehydrogenase/Δ5-4 isomerase (3βHSD) and 17α-hydroxylase/17,20-lyase cytochrome P450 (P450c17) crucial for progesterone and androgen synthesis, respectively, as well as aromatase cytochrome P450 (P450arom) that converts Δ4-androgens to estrogens. 5α-reductase is another important enzyme in equine placentas because 5α-dihydroprogesterone (DHP) sustains pregnancy in the absence of progesterone in the second half of equine pregnancy. DHP a...
A Summary of New Findings on the Biological Effects of Selenium in Selected Animal Species-A Critical Review.
International journal of molecular sciences    October 21, 2017   Volume 18, Issue 10 doi: 10.3390/ijms18102209
Hosnedlova B, Kepinska M, Skalickova S, Fernandez C, Ruttkay-Nedecky B, Malevu TD, Sochor J, Baron M, Melcova M, Zidkova J, Kizek R.Selenium is an essential trace element important for many physiological processes, especially for the functions of immune and reproductive systems, metabolism of thyroid hormones, as well as antioxidant defense. Selenium deficiency is usually manifested by an increased incidence of retention of placenta, metritis, mastitis, aborts, lowering fertility and increased susceptibility to infections. In calves, lambs and kids, the selenium deficiency demonstrates by WMD (white muscle disease), in foals and donkey foals, it is associated with incidence of WMD and yellow fat disease, and in pigs it cau...
First evidence for the presence of aquaporins in stallion sperm.
Reproduction in domestic animals = Zuchthygiene    October 21, 2017   Volume 52 Suppl 4 61-64 doi: 10.1111/rda.13059
Bonilla-Correal S, Noto F, Garcia-Bonavila E, Rodríguez-Gil JE, Yeste M, Miro J.Aquaporins (AQPs) are transmembrane proteins found in all cells and are responsible for the transport of water and small solutes. While these proteins have been found in the spermatozoa of humans, rodents, pigs and cattle, where not only do they play a role for the regulation of sperm volume but are also related with the sperm resilience to withstand freeze-thawing procedures, their presence in stallion sperm is yet to be reported. Therefore, the objectives of this work were as follows: (i) to determine whether AQP3, AQP7 and AQP11 are present in stallion sperm and (ii) to investigate whether ...
An interview with Katrin Hinrichs.
Biology of reproduction    October 19, 2017   Volume 97, Issue 5 657-659 doi: 10.1093/biolre/iox126
Hansen PJ.No abstract available
Effects of inbreeding and other systematic effects on fertility of Black Forest Draught horses in Germany.
Acta veterinaria Scandinavica    October 18, 2017   Volume 59, Issue 1 70 doi: 10.1186/s13028-017-0338-4
Müller-Unterberg M, Wallmann S, Distl O.The Black Forest Draught horse (BFDH) is an endangered German coldblood breed with its origin in the area of the Black Forest in South Germany. In this retrospective study, the influence of the inbreeding coefficient on foaling rates was investigated using records from ten breeding seasons. Due to the small population size of BFDH, the level of inbreeding is increasing and may have an effect on foaling rates.The data of the present study included all coverings reported for 1024 BFDH mares in the years 2001-2009. These mares were covered by 32 BFDH stallions from the State Stud Marbach. Data fr...
Comparison of McMaster and Mini-FLOTAC fecal egg counting techniques in cattle and horses.
Veterinary parasitology, regional studies and reports    October 10, 2017   Volume 10 132-135 doi: 10.1016/j.vprsr.2017.10.003
Dias de Castro LL, Abrahão CLH, Buzatti A, Molento MB, Bastianetto E, Rodrigues DS, Lopes LB, Silva MX, de Freitas MG, Conde MH, Borges FA.The aim of this study was to compare two fecal egg count (FEC) techniques; McMaster (McM) and Mini-FLOTAC (mF), for the detection of cattle and horse gastrointestinal nematode eggs, in different locations. Experiment 1: feces were collected from 16 cattle and FEC was performed individually, using mF with the sensitivity of 5 eggs per gram of feces (EPG) and McM with a sensitivity of 50 EPG at Empresa de Pesquisa Agropecuária de Minas Gerais - EPAMIG and the Laboratory of Parasitic Diseases of the University of Parana - LDP/UFPR. Experiment 2: Fecal samples from 30 horses were analyzed with mF...
Reliability of histopathological examination and immunohistochemistry of a single biopsy for evaluation of endometrial health in Icelandic mares.
Folia histochemica et cytobiologica    October 10, 2017   Volume 55, Issue 3 168-175 doi: 10.5603/FHC.a2017.0017
Sikora M, Nowak M, Racheniuk H, Wojtysiak K, Kozdrowski R.Endometritis and endometrosis have been and still are the major reasons for infertility in mares. The diagnosis of endometritis can be based on cytology and microbiology, but endometrial biopsy is still the only way to diagnose endometrosis in the mare. Our study attempted to determine if a single biopsy using his-topathology and immunohistochemistry is sufficient to ascertain reasons for infertility in Icelandic mares. The objectives of this study were to examine the relationship between deviations in endometrial biopsies in terms of prostaglandin-endoperoxide synthase-2 (PTGS-2) and fibronec...
A comparison of progesterone assays for determination of peripheral pregnane concentrations in the late pregnant mare.
Theriogenology    October 7, 2017   Volume 106 127-133 doi: 10.1016/j.theriogenology.2017.10.002
Wynn MAA, Esteller-Vico A, Legacki EL, Conley AJ, Loux SC, Stanley SD, Curry TE, Squires EL, Troedsson MH, Ball BA.During the latter half of gestation in mares, there is a complex milieu of pregnanes in peripheral blood. Progesterone concentrations are often assessed by immunoassay during late gestation as a measure of pregnancy well-being; however, interpretation of results is complicated by the numerous cross-reacting pregnanes present in high concentrations during late gestation. Further, many mares are supplemented with an exogenous progestin, altrenogest, which may also cross-react with existing assays and further confound interpretation. The objectives of this study were: 1) to compare differences in...
Equine chlamydiosis-An emerging infectious disease requiring a one health surveillance approach.
Zoonoses and public health    October 5, 2017   Volume 65, Issue 1 218-221 doi: 10.1111/zph.12391
Taylor KA, Durrheim D, Heller J, O'Rourke B, Hope K, Merritt T, Freeman P, Chicken C, Carrick J, Branley J, Massey P.Psittacosis is a rare but potentially fatal zoonosis caused by Chlamydia psittaci, an organism that is typically associated with bird contact. However C. psittaci is capable of infecting other non-avian hosts, such as horses, sheep, cattle and goats. Stud staff and veterinarians have significant exposure to parturient animals and reproductive materials in their routine work. To investigate the zoonotic potential associated with the emergence of C. psittaci as an abortifacient agent in horses, we established a programme of joint human and animal surveillance in a sentinel horse-breeding regio...
Equine Arteritis Virus Elicits a Mucosal Antibody Response in the Reproductive Tract of Persistently Infected Stallions.
Clinical and vaccine immunology : CVI    October 5, 2017   Volume 24, Issue 10 doi: 10.1128/CVI.00215-17
Carossino M, Wagner B, Loynachan AT, Cook RF, Canisso IF, Chelvarajan L, Edwards CL, Nam B, Timoney JF, Timoney PJ, Balasuriya UBR.Equine arteritis virus (EAV) has the ability to establish persistent infection in the reproductive tract of the stallion (carrier) and is continuously shed in its semen. We have recently demonstrated that EAV persists within stromal cells and a subset of lymphocytes in the stallion accessory sex glands in the presence of a significant local inflammatory response. In the present study, we demonstrated that EAV elicits a mucosal antibody response in the reproductive tract during persistent infection with homing of plasma cells into accessory sex glands. The EAV-specific immunoglobulin isotypes i...
Serum anti-Müllerian hormone dynamics in mares following immunocontraception with anti-zona pellucida or -GnRH vaccines.
Theriogenology    October 5, 2017   Volume 106 214-220 doi: 10.1016/j.theriogenology.2017.10.004
Joonè CJ, Schulman ML, Fosgate GT, Claes ANJ, Gupta SK, Botha AE, Human A, Bertschinger HJ.Circulating anti-Müllerian hormone concentration (AMH) is positively correlated to the number of small growing follicles in the mare and may reflect ovarian function. Dynamics of AMH during immunocontraception have not previously been investigated. This study aimed to compare serum AMH in mares following treatment with native porcine zona pellucida (pZP), recombinant pZP3 and pZP4 (reZP) or gonadotrophin releasing hormone (GnRH) vaccines, and saline-treated controls. Stored sera collected during two previous studies examining ovarian activity in mares during zona pellucida (ZP) or GnRH immuno...
Stallion spermatozoa surviving freezing and thawing experience membrane depolarization and increased intracellular Na.
Andrology    October 3, 2017   Volume 5, Issue 6 1174-1182 doi: 10.1111/andr.12419
Ortega Ferrusola C, Anel-López L, Ortiz-Rodriguez JM, Martin Muñoz P, Alvarez M, de Paz P, Masot J, Redondo E, Balao da Silva C, Morrell JM....In order to gain insight of the modifications that freezing and thawing cause to the surviving population of spermatozoa, changes in the potential of the plasma membrane (Em) and intracellular Na content of stallion spermatozoa were investigated using flow cytometry. Moreover, caspase 3 activity was also investigated and the functionality of the Na -K ATPase pump was investigated before and after freezing and thawing. Cryopreservation caused a significant (p < 0.001) increase in the subpopulation of spermatozoa with depolarized sperm membranes, concomitantly with an increase (p < 0.0...
Alpha lipoic acid (ALA) effects on developmental competence of equine preantral follicles in short-term culture.
Theriogenology    September 27, 2017   Volume 105 169-173 doi: 10.1016/j.theriogenology.2017.09.023
Gomes RG, Silva CB, González SM, Oliveira RL, Max MC, Lisboa LA, Barreiros TRR, Santos MM, Sarapião FD, Gastal EL, Seneda MM.The effect of different concentrations of alpha lipoic acid (ALA) on the development and morphology of preantral follicles, as well as the proliferative activity of granulosa cells, was assessed after short-term culture. Ovaries (n = 5) of five seasonal anestrous mares were harvested in a local abattoir. At the laboratory, nine ovarian fragments (5 × 5 × 1 mm) from each animal were used. One fragment was immediately fixed in Bouin and subjected to histological and immunohistochemistry (proliferating cell nuclear antigen, PCNA) analyses (noncultured group; D0 = day 0). The other eigh...
Do olfactory behaviour and marking responses of Konik polski stallions to faeces from conspecifics of either sex differ?
Behavioural processes    September 27, 2017   Volume 155 38-42 doi: 10.1016/j.beproc.2017.09.015
Jezierski T, Jaworski Z, Sobczyńska M, Ensminger J, Górecka-Bruzda A.The aim of the study was to assess whether adult stallions differentiate their olfactory and marking behaviour towards the excreta of their potential male rivals and mares. Four Konik polski stallions were individually exposed simultaneously to their own and each others' faeces as well as faeces of mares in oestrus vs. dioestrus. Five series of 30min observation sessions were conducted in a round pen each on two consecutive days from March to July, totaling in 5h observation per stallion. Stallions sniffed and displayed flehmen reaction towards mares' faeces significantly longer (P<0.01) th...
Phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) and Janus kinase/signal transducer and activator of transcription (JAK/STAT) follicular signalling is conserved in the mare ovary.
Reproduction, fertility, and development    September 26, 2017   Volume 30, Issue 4 624-633 doi: 10.1071/RD17024
Hall SE, Upton RMO, McLaughlin EA, Sutherland JM.The mare ovary is unique in its anatomical structure; however, the signalling pathways responsible for physiological processes, such as follicular activation, remain uncharacterised. This provided us with the impetus to explore whether signalling molecules from important folliculogenesis pathways, phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) and Janus kinase/signal transducer and activator of transcription (JAK/STAT), are conserved in the mare ovary. Messenger RNA expression of six genes important in follicle development was measured using quantitative polymerase chain reaction and pr...
Morphological evaluation of Day 8 embryos developed during induced aluteal cycles in the mare.
Theriogenology    September 25, 2017   Volume 105 178-183 doi: 10.1016/j.theriogenology.2017.09.029
Leisinger CA, Medina V, Markle ML, Paccamonti DL, Pinto CRF.A novel in vivo model utilizing serial administrations of PGF2α to induce aluteal cycles in the mare was used to evaluate the effects of progesterone-deprivation on the morphology of in vivo preimplantation embryos. We hypothesized that equine embryos produced during induced aluteal cycles (AL) would be developmentally affected, characterized by earlier embryo stage at collection, smaller embryo diameter, and lower quality grade, compared with those collected on the same day post-ovulation from control cycles during diestrus (high progesterone; > 4 ng/mL). Seven cyclic mares with a median ...
A Brief Account of the Discovery of the Fetal/Placental Unit for Estrogen Production in Equine and Human Pregnancies: Relation to Human Medicine.
The Yale journal of biology and medicine    September 25, 2017   Volume 90, Issue 3 449-461 
Raeside JI.The role of steroids in human medicine is well recognized, but the major contributions made by the large domestic animals as a source of material in the discovery, isolation, and determination of the structure of the steroid hormones is less well appreciated. After a brief reminder of the early efforts to obtain a reliable source of steroids for clinical use, the narrative here is to outline one example where success was ultimately achieved for estrogen replacement therapy. Whereas knowledge of the high concentrations of estrogens in urine of pregnant women and mares dates from the late 1920s,...
An improved vitrification protocol for equine immature oocytes, resulting in a first live foal.
Equine veterinary journal    September 21, 2017   Volume 50, Issue 3 391-397 doi: 10.1111/evj.12747
Ortiz-Escribano N, Bogado Pascottini O, Woelders H, Vandenberghe L, De Schauwer C, Govaere J, Van den Abbeel E, Vullers T, Ververs C, Roels K....The success rate for vitrification of immature equine oocytes is low. Although vitrified-warmed oocytes are able to mature, further embryonic development appears to be compromised. Objective: The aim of this study was to compare two vitrification protocols, and to examine the effect of the number of layers of cumulus cells surrounding the oocyte during vitrification of immature equine oocytes. Methods: Experimental in vitro and in vivo trials. Methods: Immature equine oocytes were vitrified after a short exposure to high concentrations of cryoprotective agents (CPAs), or a long exposure to l...
Comparison of physical body growth and metabolic and reproductive endocrine functions between north and south climates of Japan in trained Thoroughbred yearling horses.
Journal of equine science    September 20, 2017   Volume 28, Issue 3 77-86 doi: 10.1294/jes.28.77
Tangyuenyong S, Sato F, Nambo Y, Murase H, Endo Y, Tanaka T, Nagaoka K, Watanabe G.This study aimed to compare body growth, metabolic, and reproductive hormonal changes in trained Thoroughbred yearling horses under different climate conditions with and without light supplementation (LS). Thoroughbred yearlings raised at research centers of the Japan Racing Association in Hokkaido (north) or Miyazaki (south) were divided into control and LS groups. In the LS groups, 44 colts and 47 fillies from Hokkaido and 11 colts and 11 fillies from Miyazaki were exposed to LS with an extended photoperiod of 14.5 hr of daylight and 9.5 hr of darkness. One week before and once a month after...
Exposure to follicular fluid during oocyte maturation and oviductal fluid during post-maturation does not improve in vitro embryo production in the horse.
Zygote (Cambridge, England)    September 20, 2017   Volume 25, Issue 5 612-630 doi: 10.1017/S096719941700048X
Douet C, Parodi O, Martino NA, Lacalandra GM, Nicassio M, Reigner F, Deleuze S, Dell'Aquila ME, Goudet G.Most wild equids and many domestic horse breeds are at risk of extinction, so there is an urgent need for genome resource banking. Embryos cryopreservation allows the preservation of genetics from male and female and is the fastest method to restore a breed. In the equine, embryo production in vitro would allow the production of several embryos per cycle. Intracytoplasmic sperm injection (ICSI) is used to generate horse embryos, but it requires expensive equipment and expertise in micromanipulation, and blastocyst development rates remain low. No conventional in vitro fertilization (IVF) techn...
Lipidomics of equine amniotic fluid: Identification of amphiphilic (O-acyl)-ω-hydroxy-fatty acids.
Theriogenology    September 20, 2017   Volume 105 120-125 doi: 10.1016/j.theriogenology.2017.09.012
Wood PL, Ball BA, Scoggin K, Troedsson MH, Squires EL.Amniotic fluid is essential for the growth and maturation of the fetus prior to parturition. While our knowledge of human amniotic fluid is extensive, current data for equine amniotic fluid is limited. We therefore undertook a detailed lipidomics analysis of equine amniotic fluid. Using a non-targeted high-resolution mass spectrometric lipidomics analysis of equine amniotic fluid, we were able to characterize a diverse array of individual lipids. This non-biased analytical approach detected, for the first time, the presence of (O-acyl)-ω-hydroxy-fatty acids (OAHFA) with up to 52 carbon chain ...
Measurement of urinary concentrations of the mycotoxins zearalenone and sterigmatocystin as biomarkers of exposure in mares.
Reproduction in domestic animals = Zuchthygiene    September 18, 2017   Volume 53, Issue 1 68-73 doi: 10.1111/rda.13054
Takagi M, Uno S, Kokushi E, Sato F, Wijayagunawardane M, Fink-Gremmels J.Mycotoxins may affect animal health, including reproduction. Little is known about the clinical relevance of exposure of horses to contaminated feed. This study aimed at (i) monitoring the levels of the mycotoxins zearalenone (ZEN), with its metabolites α- and β-zearalenol (α- and β-ZOL), and sterigmatocystin (STC) in urine samples from thoroughbred mares in Japan and (ii) relating these findings to the potential effects on reproductive efficacy of breeding mares. Sixty-three urine samples of breeding mares from 59 breeding farms were used. Urine samples and reproductive records were colle...
Pre-ovulatory follicle affects corpus luteum diameter, blood flow, and progesterone production in mares.
Animal reproduction science    September 8, 2017   Volume 187 1-12 doi: 10.1016/j.anireprosci.2017.09.003
Ishak GM, Bashir ST, Gastal MO, Gastal EL.Color Doppler ultrasonography was used to study the temporal relationships between pre-ovulatory follicle (POF) and corpus luteum (CL) diameter and blood flow, with systemic progesterone (P4) concentration during two transitional ovulatory seasons in mares. Variables of POF and CL/P4 were evaluated for 6days before and 17days after ovulation, respectively. Evaluations were performed during two consecutive estrous cycles in spring and fall seasons, and during the last estrous cycle of the season. There were significant correlations among POF and CL variables, and P4 concentration that ranged fr...
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