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Animal reproduction science.

Periodical
Reproductive Medicine
Veterinary Medicine
Zoology
Animals
Reproduction
Publisher:
Elsevier Scientific Pub. Co.
Frequency: Twenty eight no. a year, 2008-
Country: Netherlands
Language: English
Start Year:1978 -
ISSN:
0378-4320 (Print)
1873-2232 (Electronic)
0378-4320 (Linking)
Impact Factor
2.2
2022
NLM ID:7807205
(DNLM):A31035000(s)
(OCoLC):04122595
Coden:ANRSDV
LCCN:sc 80000012
Classification:W1 AN228V
Past, present and future of ICSI in livestock species.
Animal reproduction science    January 19, 2022   Volume 246 106925 doi: 10.1016/j.anireprosci.2022.106925
Briski O, Salamone DF.During the past 2 decades, intracytoplasmic sperm injection (ICSI) has become a routine technique for clinical applications in humans. The widespread use among domestic species, however, has been limited to horses. In horses, ICSI is used to reproduce elite individuals and, as well as in humans, to mitigate or even circumvent reproductive barriers. Failures in superovulation and conventional in vitro fertilization (IVF) have been the main reason for the use of this technology in horses. In pigs, ICSI has been successfully used to produce transgenic animals. A series of factors have resulted in...
Advances in sperm cryopreservation in farm animals: Cattle, horse, pig and sheep.
Animal reproduction science    December 3, 2021   Volume 246 106904 doi: 10.1016/j.anireprosci.2021.106904
Yánez-Ortiz I, Catalán J, Rodríguez-Gil JE, Miró J, Yeste M.Sperm cryopreservation is one of the most important procedures in the development of biotechnologies for assisted reproduction. In some farm animals, the use of cryopreserved sperm has so many benefits for which relevance has become more evident in recent decades. Values for post-thaw sperm quality, however, are variable among species and within individuals of the same species. There is no standardized methodology for each of the stages of the cryopreservation procedure (andrological examination, semen collection, dilution, centrifugation, resuspension of the pellet with the freezing medium, p...
Preliminary study of the contraceptive effect of a self-assembling intrauterine device (iUPODs) in mares maintained in a paddock with a fertile stallion.
Animal reproduction science    October 30, 2021   Volume 235 106881 doi: 10.1016/j.anireprosci.2021.106881
Hoopes KH, Gradil CM, Vanderwall DK, Mason HM, Sarnecky BA, Davies CJ.There is an urgent need for practical methods of population control (i.e., contraception and/or sterilization) for free-roaming (i.e., "wild" or "feral") horses and burros on Western Public Lands in the United States. The objective of this study was to evaluate the contraceptive efficacy of a novel self-assembling three-part polymer-coated magnetic intrauterine device termed as an intrauterine POD (self-assembling; iUPOD) when there are natural breeding conditions when iUPOD use was managed by veterinary professionals with no prior experience with the device. Six mares were administered an iUP...
Modification of O-ring intrauterine devices (IUDs) in mares: Contraception without estrus suppression.
Animal reproduction science    September 25, 2021   Volume 234 106864 doi: 10.1016/j.anireprosci.2021.106864
Lyman CC, Baldrighi JM, Anderson CO, Germaine SS, Kane AJ, Holyoak GR.A significant welfare concern for those within the horse industry is that of the overpopulation in free-roaming horses in the western US. The goal of the present study was to identify a humane and inexpensive means of providing contraception and reducing pregnancy rates in a way that could be applied to free-roaming horses on the western U.S. open range lands. A series of studies was conducted utilizing the previously-described silastic O-ring intrauterine devices (IUDs). Throughout these studies pregnancy was successfully prevented in mares with an O-ring IUD, but collective retention rates f...
A stallion spermatozoon’s journey through the mare’s genital tract: In vivo and in vitro aspects of sperm capacitation.
Animal reproduction science    September 14, 2021   Volume 246 106848 doi: 10.1016/j.anireprosci.2021.106848
Maitan P, Bromfield EG, Stout TAE, Gadella BM, Leemans B.Conventional in vitro fertilization is not efficacious when working with equine gametes. Although stallion spermatozoa bind to the zona pellucida in vitro, these gametes fail to initiate the acrosome reaction in the vicinity of the oocyte and cannot, therefore, penetrate into the perivitelline space. Failure of sperm penetration most likely relates to the absence of optimized in vitro fertilization media containing molecules essential to support stallion sperm capacitation. In vivo, the female reproductive tract, especially the oviductal lumen, provides an environmental milieu that appropriate...
Cooled storage of semen from livestock animals (part I): boar, bull, and stallion.
Animal reproduction science    August 12, 2021   Volume 246 106822 doi: 10.1016/j.anireprosci.2021.106822
Wiebke M, Hensel B, Nitsche-Melkus E, Jung M, Schulze M.This review is part of the Festschrift in honor of Dr. Duane Garner and provides an overview of current techniques for cooled storage of semen from livestock animals. The first part describes the current state of the art of liquid semen preservation in boars, bulls, and stallions, including the diluents, use of additives, processing, temperature, and cooling of semen. The species-specific physiology and varying extents of cold shock sensitivity are taken into consideration. In addition, factors influencing the quality of cooled-stored semen are discussed. Methods, trends, and the most recent a...
Culture protocols for horse embryos after ICSI: Effect of myo-inositol and time of media change.
Animal reproduction science    July 27, 2021   Volume 233 106819 doi: 10.1016/j.anireprosci.2021.106819
Brom-de-Luna JG, Salgado RM, Felix MR, Canesin HS, Stefanovski D, Diaw M, Hinrichs K.In vitro production of horse embryos via intracytoplasmic sperm injection (ICSI) is a useful clinical and research technique. Current rates of blastocyst production are typically sub-optimal, and few methods to increase the rate of equine blastocyst development have been reported. Factors that might improve blastocyst production in a horse embryo culture system were explored. Myo-inositol is found in the horse oviduct and improves blastocyst development in other species, thus Experiment 1 was conducted to assess the effect of 10 mM myo-inositol added to Day 0-5 embryo culture medium, using hor...
An integrated overview on the regulation of sperm metabolism (glycolysis-Krebs cycle-oxidative phosphorylation).
Animal reproduction science    July 14, 2021   Volume 246 106805 doi: 10.1016/j.anireprosci.2021.106805
Peña FJ, Ortiz-Rodríguez JM, Gaitskell-Phillips GL, Gil MC, Ortega-Ferrusola C, Martín-Cano FE.An overview of the sperm metabolism is presented; using the stallion as a model we review glycolysis, Krebs Cycle and oxidative phosphorylation, paying special attention to the interactions among them. In addition, metabolism implies a series of coordinated oxidation-reduction reactions and in the course of these reactions reactive oxygen species (ROS) and reactive oxoaldehydes are produced ; the electron transport chain (ETC) in the mitochondria is the main source of the anion superoxide and hydrogen peroxide, while glycolysis produces 2-oxoaldehydes such as methylglyoxal as byproducts; due t...
An intrauterine device with potential to control fertility in feral equids.
Animal reproduction science    June 17, 2021   Volume 231 106795 doi: 10.1016/j.anireprosci.2021.106795
Gradil C, Joone C, Haire T, Fowler B, Zinchuk J, Davies CJ, Ball B.Fertility control of feral equids is difficult. A 4-month pilot study was conducted with a hormone-free intrauterine device (iUPOD). There was evaluation of i) device retention; ii) contraceptive efficacy; iii) fertility following device removal; iv) effects of device on estrous cycle periodicity and; v) abundance of biofilm on devices after removal from the uterus. The iUPODs were inserted trans-cervically in eight mares at random stages of the estrous cycle. Mares were confined in a paddock with a stallion the following day and remained with the stallion for 120 days. Transabdominal detectio...
Evaluation of mare endometrial cytology using the novel cytotape technique.
Animal reproduction science    May 18, 2021   Volume 230 106770 doi: 10.1016/j.anireprosci.2021.106770
Ibrahim M, Ferrer MS, Ellerbrock RE, Rollin E.The cytobrush is considered the method of choice to obtain endometrial samples. Rigid brush fibers, however, may induce endometrial irritation and bleeding, or cell fragmentation, decreasing quality and diagnostic value of the samples. It was hypothesized that samples collected using a novel cytotape would provide sample smears of greater quality and less blood contamination than the cytobrush. Endometrial samples were collected with a cytotape and a cytobrush from ten mares without endometritis. Endometritis was then induced with artificial insemination, and samples were again collected 6 h a...
Serum amyloid A, Serum Amyloid A1 and Haptoglobin in pregnant mares and their fetuses after experimental induction of placentitis.
Animal reproduction science    May 11, 2021   Volume 229 106766 doi: 10.1016/j.anireprosci.2021.106766
Boakari YL, Esteller-Vico A, Loux S, El-Sheikh Ali H, Fernandes CB, Dini P, Scoggin KE, Cray C, Ball BA.Serum amyloid A (SAA) and Haptoglobin (Hp) are acute phase proteins, produced during inflammation, such as placentitis. In horses, SAA and SAA1 are protein coding genes. Objectives were to analyze SAA and Hp concentrations and relative abundance of SAA, SAA1 and Hp mRNA transcript in maternal and fetal tissues after experimental induction of placentitis or mares of a control group. Serum Amyloid A family proteins were in marked abundance in the stroma of the endometrium and chorioallantois associated with inflammatory cells. Maternal plasma SAA concentrations were greater (P = 0.01) in mares w...
Effect of maternal overnutrition on predisposition to insulin resistance in the foal: Maternal parameters and foal pancreas histoarchitecture.
Animal reproduction science    February 12, 2021   Volume 227 106720 doi: 10.1016/j.anireprosci.2021.106720
Bradbery AN, Coverdale JA, Hartz CJ, Millican AA, Goehring MS, Fikes KK, Picking E, Hammer CJ, Dunlap KA, Cardoso RC, Wickersham TA, Leatherwood JL....Results from previous studies indicate that maternal overnutrition during late gestation predisposes foals to metabolic disease, however, specific mechanisms resulting in disease remain unknown. Quarter Horse mares (n = 16), were randomly assigned to dietary treatments, beginning on gestational day 235, and consisted of a control group (CON- diet meeting nutrient requirement; n = 8) or an overfed diet (HIGH; n = 8) where mares received an additional 40 % above CON. On gestational days 285 and 315, an intravenous glucose tolerance test (FSIGTT) was conducted. Following parturition, foals were s...
Changes in circulating concentrations of testosterone and estrone sulfate after human chorionic gonadotropin administration and subsequent to castration of 2-year-old stallions.
Animal reproduction science    December 3, 2020   Volume 225 106670 doi: 10.1016/j.anireprosci.2020.106670
Esteller-Vico A, Ball BA, Bridges JW, Hughes SE, Squires EL, Troedsson MHT.Reproductive steroids testosterone (T) and estrone sulfate (E1S) are used as diagnostic markers for cryptorchidism in horses. The human chorionic gonadotropin (hCG) stimulation test is used as a diagnostic aid because administration of this hormone results in greater incremental differences in circulating steroid concentrations. Thoughts regarding optimal sampling times following hCG administration, however, are inconsistent. Additionally, determination of half-life of these steroids is important in postsurgical samples to confirm complete removal of testicular tissue. Objectives of this study...
Optimal frame rate when there were stallion sperm motility evaluations and determinations for kinematic variables using CASA-Mot analysis in different counting chambers.
Animal reproduction science    November 2, 2020   Volume 223 106643 doi: 10.1016/j.anireprosci.2020.106643
Gacem S, Bompart D, Valverde A, Catalán J, Miró J, Soler C.This study was conducted to determine optimum image capture frame rates (FRO) when there was evaluation of different types of counting chambers used for CASA-Mot determinations of stallion sperm motility. Sperm VCL was determined at frame rates of 25-250 f/s in: 1) Spermtrack® (Spk) 10 and 20 chambers (drop displacement-type chambers 10 and 20 μm-deep respectively; and 2) ISAS®D4C10, ISAS®D4C20 (10 and 20 μm-deep respectively) and ISAS®D4C20 L (20 μm-deep) capillary loaded chambers. Values for different sperm kinematic variables were determined using each chamber at 250 f/s, whi...
Effect of season on individual stallion semen characteristics.
Animal reproduction science    November 1, 2020   Volume 223 106641 doi: 10.1016/j.anireprosci.2020.106641
Crespo F, Wilson R, Díaz-Jimenez M, Consuegra C, Dorado J, Barrado BG, Gosálvez J, Smit RL, Hidalgo M, Johnston S.Horses are long-day seasonal breeding animals, however, with modern stallion reproductive management it is important for collection of semen during periods that are not part of the traditional breeding season. This study was conducted to examine variation in the seminal characteristics of individual stallions in Avila, Spain during 1 year with a particular emphasis on sperm DNA fragmentation. Semen was collected twice per season from a total of 20 stallions. There was a marked seasonal effect on all seminal characteristics, with the greatest on progressive motility, % membrane integrity and le...
Fertilizing capacity of vitrified stallion sperm assessed utilizing heterologous IVF after different semen warming procedures.
Animal reproduction science    October 15, 2020   Volume 223 106627 doi: 10.1016/j.anireprosci.2020.106627
Consuegra C, Crespo F, Dorado J, Diaz-Jimenez M, Pereira B, Sánchez-Calabuig MJ, Beltrán-Breña P, Pérez-Cerezales S, Rizos D, Hidalgo M.The aim of this study was to evaluate the fertilizing capacity of frozen or vitrified stallion sperm after assessing different warming procedures. In Experiment 1, different warming procedures were compared after sperm vitrification: immersion in extender at 43 °C (C), or in a water bath at 37 °C/30 s (W37), 43 °C/10 s (W43) or 60 °C/5 s (W60). With the W60 treatment, there were greater values (P < 0.05) for VCL (83.93 ± 3.6 μm/s) and ALH (3.00 ± 0.2 μm) than freezing and with the C group, and greater values (P < 0.001) for PM (35.33 ± 2.5 %) than with the W43 treatment. In Experiment...
Oral misoprostol does not hasten oviductal transport of day-5 horse embryos.
Animal reproduction science    October 14, 2020   Volume 223 106626 doi: 10.1016/j.anireprosci.2020.106626
Checura CM, Momont HW, Castañeira C, Flores-Bragulat A, Losinno L.In horses, prostaglandin E (PGE) is produced by embryos around Day 5 post-ovulation; PGE functions directly at the oviduct promoting embryo transport into the uterus. Non-surgical collection of horse embryos for cryopreservation is recommended at Day 6.5-7 post-ovulation. It was proposed that misoprostol administered orally will hasten oviductal transport of horse embryos. In Experiment 1 (n = 15) there was comparison of time of embryo recovery (Day 6 and 6.5 post-ovulation) from mares administered misoprostol (Day 5 and 5.5) orally to that of untreated mares. On Day 6, embryo collections were...
A review of in vivo and in vitro studies of the mare endometrium.
Animal reproduction science    September 16, 2020   Volume 222 106605 doi: 10.1016/j.anireprosci.2020.106605
Thompson RE, Premanandan C, Pukazhenthi BS, Whitlock BK.The inner layer of the uterus, the endometrium, is responsible and necessary for many reproductive functions. Normal reproductive cyclicity, maternal recognition of pregnancy, maternal interaction with the embryo, and interaction of the reproductive tract with pathogens are dependent on the endometrium. Although most studies have been conducted in vivo using live animals, recent advances in in vitro approaches could facilitate future research in a laboratory setting with minimal effect on animals. Many reproductive studies have been performed in vivo and in vitro in equids, but new in vitro me...
Optimization of cryopreservation protocols for cooled-transported stallion semen.
Animal reproduction science    August 20, 2020   Volume 221 106581 doi: 10.1016/j.anireprosci.2020.106581
Ferrer MS, Canisso IF, Ellerbrock RE, Podico G, Lister BN, Hurley DJ, Kline K, Palomares RA.Freezing cooled-transported semen allows veterinarians and breeders to collect and process the semen of stallions on farm, and then ship the semen to a semen freezing center. There, however, is a lack of standardization of shipping and freezing protocols. The objectives were to optimize and simplify protocols to freeze cooled-shipped semen. In Experiment 1, cooled-transported semen was centrifuged at room temperature or 5 °C before freezing. Sperm variables (motility, membrane integrity, acrosome integrity, membrane fluidity) were evaluated before and after freezing. Centrifugation temperat...
Chemotactic selection of frozen-thawed stallion sperm improves sperm quality and heterologous binding to oocytes.
Animal reproduction science    August 20, 2020   Volume 221 106582 doi: 10.1016/j.anireprosci.2020.106582
Dominguez EM, Moreno-Irusta A, Rodriguez MB, Salamone DF, de Arruda RP, Losinno L, Giojalas LC.The successful use of assisted reproduction techniques (ART) depends in part on the sperm physiological status. Several sperm selection procedures have been applied to improve quality of sperm population when using the ART. There has previously been development of a Sperm Selection Assay (SSA) for humans which is based on the attraction of capacitated sperm by chemotaxis towards progesterone (P), resulting in an enriched sperm population with an optimal physiological status similar to capacitated spermatozoa, with these cells having very little DNA fragmentation and optimal concentrations of r...
Effect of MnTBAP on in vitro capacitation of frozen-thawed stallion sperm.
Animal reproduction science    August 5, 2020   Volume 221 106570 doi: 10.1016/j.anireprosci.2020.106570
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...
Metagenomic analysis of bacteria in stallion semen.
Animal reproduction science    August 1, 2020   Volume 221 106568 doi: 10.1016/j.anireprosci.2020.106568
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...
High-throughput sperm assay using label-free microscopy: morphometric comparison between different sperm structures of boar and stallion spermatozoa.
Animal reproduction science    May 23, 2020   Volume 219 106509 doi: 10.1016/j.anireprosci.2020.106509
Rubessa M, Feugang JM, Kandel ME, Schreiber S, Hessee J, Salerno F, Meyers S, Chu I, Popescu G, Wheeler MB.The capacity for microscopic evaluation of sperm is useful for assisted reproductive technologies (ART), because this can allow for specific selection of sperm cells for in vitro fertilization (IVF). The objective of this study was to analyze the same sperm samples using two high-resolution methods: spatial light interference microscopy (SLIM) and atomic force microscopy (AFM) to determine if with one method there was more timely and different information obtained than the other. To address this objective, there was evaluation of sperm populations from boars and stallions. To the best of our k...
Effect of oral urea supplementation on the endometrial transcriptome of mares.
Animal reproduction science    April 18, 2020   Volume 216 106464 doi: 10.1016/j.anireprosci.2020.106464
Linhares Boakari Y, El-Sheikh Ali H, Dini P, Loux S, Barbosa Fernandes C, Esteller-Vico A, Scoggin K, Lawrence L, Ball B.An intravenous large dose of protein led to an increased blood urea nitrogen (BUN), resulting in a lesser uterine pH and altered uterine gene expression in mares. The objective of the present study was to evaluate effects of a more physiological methodology to increase BUN on the endometrium of mares. Mares were fed hay and a treatment or control diet (n = 11 mares/treatment) in a crossover design starting at time of ovulation detection (D0) and continuing until D7. Mares of the treated group were fed urea (0.4 g/kg BW) with sweet feed and molasses, and those of the control group were fe...
Effect of relaxin on semen quality variables of cryopreserved stallion semen.
Animal reproduction science    April 2, 2020   Volume 216 106351 doi: 10.1016/j.anireprosci.2020.106351
Elkhawagah AR, Nervo T, Poletto M, Martino NA, Gallo D, Bertero A, Vincenti L.The aim of the study was to ascertain effects of different concentrations of relaxin added to extender medium during the pre-freezing incubation periods on quality variables of stallion frozen-thawed spermatozoa. Semen samples collected from three stallions were filtered, diluted with skim milk, and centrifuged at 600g for 10 min. Sperm pellets were suspended in BotuCrio freezing medium to a final concentration of 50 × 10 sperm/mL. The diluted semen was divided into five experimental groups supplemented with 0 (control), 12.5, 25, 50, or 100 ng/mL of relaxin. The semen samples were transferre...
The role of seminal plasma in the liquid storage of spermatozoa.
Animal reproduction science    January 24, 2020   Volume 220 106290 doi: 10.1016/j.anireprosci.2020.106290
Höfner L, Luther AM, Waberski D.Ongoing progress in proteomic characterization of seminal plasma has stimulated research on the identification of biomarkers for male fertility and sperm preservability. So far, many studies have evaluated the benefits of reconstituting cryopreserved or sex-sorted semen with seminal plasma. Less information is available about the effect of remaining or added seminal plasma in liquid preserved semen. The interaction between seminal plasma and spermatozoa is species -specific, and within species often complex and ambiguous. This article aims to review the action of seminal plasma on sperm functi...
Effects of media and promoters on different lipid peroxidation assays in stallion sperm.
Animal reproduction science    November 1, 2019   Volume 211 106199 doi: 10.1016/j.anireprosci.2019.106199
Ghosh S, Serafini R, Love CC, Teague SR, Hernández-Avilés C, LaCaze KA, Varner DD.Effects of different media and promoters on lipid peroxidation (LPO) in viable stallion sperm have not been reported. Aims of this study were to determine effects of three media (INRA-96™, Equipro CoolGuard™, and Biggers, Whitten and Whittingham [BWW]), and promoters (iron sulfate-Fe; ultraviolet light-UV; or control-no exposure to promoters) on viable sperm LPO using four different flow cytometric assays (i.e., BODIPY, Liperfluo, 4-hydroxylnonenal [4HNE], malonaldehyde [MDA]). Significant media x promoter interactions were detected using the Liperfluo, 4HNE, and MDA assays (P <  0.05)...
Ejaculated compared with epididymal stallion sperm vitrification.
Animal reproduction science    October 22, 2019   Volume 211 106205 doi: 10.1016/j.anireprosci.2019.106205
Álvarez C, González N, Luño V, Gil L.The aim of this study was to evaluate the effect of trehalose and lactose extenders on ejaculated and epididymal stallion sperm vitrification. Ejaculated semen samples were collected from seven fertile stallions, and cauda epididymis samples were collected from ten stallion carcasses after slaughter. Both the ejaculated and the epididymis samples were diluted and vitrified using INRA 96® and bovine serum albumin as well as trehalose or lactose. As a control, ejaculated and epididymal samples were collected and frozen using the conventional method. Vitrification was performed by immersing sper...
Characterization of abortion, stillbirth and non-viable foals homozygous for the Warmblood Fragile Foal Syndrome.
Animal reproduction science    October 17, 2019   Volume 211 106202 doi: 10.1016/j.anireprosci.2019.106202
Aurich C, Müller-Herbst S, Reineking W, Müller E, Wohlsein P, Gunreben B, Aurich J.Warmblood fragile foal syndrome (WFFS) is a monogenetic defect with autosomal recessive inheritance. The WFFS homozygosity is non-compatible with extra-uterine life. Although as many as 15% of Warmblood horses are WFFS carriers, there has been little veterinary focus on this condition. The aim of this study was to determine outcomes and symptoms of clinical signs and pathological abnormalities during pregnancies when there were WFFS homozygous foetuses. Diagnostic material of 15 abortion or stillbirth cases with suspected diagnosis of WFFS was available for this study. Additionally, there were...
Genetic parameters and estimated breeding values for traits of raw and frozen-thawed semen in German Warmblood stallions.
Animal reproduction science    September 23, 2019   Volume 210 106194 doi: 10.1016/j.anireprosci.2019.106194
Greiser T, Sieme H, Martinsson G, Distl O.Objectives of the present study were to estimate genetic parameters for frozen-thawed semen traits of 271 fertile German Warmblood stallions and genetic correlations with raw semen quality traits. Semen samples were collected from stallions utilized for semen collection and artificial insemination (AI) on the Lower Saxon National Stud Celle and the North Rhine-Westphalian National Stud Warendorf. Semen quality variables were analyzed in 63,972 raw (gel-free volume, concentration, progressive motility, number of sperm) and 3681 frozen-thawed samples (motility, DNA fragmentation index (DFI), non...