scholarly journals Short-term liquid storage of ram semen in various extenders

1970 ◽  
Vol 48 (4) ◽  
pp. 717-723
Author(s):  
N. Maksimović ◽  
A. Milovanović ◽  
T. Barna ◽  
V. Caro Petrović ◽  
V. Pantelić ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Morphology ◽  
Sperm Quality ◽  
Animal Husbandry ◽  
Field Analysis ◽  
Blue Staining ◽  
Computer Assisted ◽  
Short Term ◽  
Sperm Analysis ◽  
Liquid Storage

The aim of this study was to investigate the effects of three extenders on ram sperm quality after short-term liquid storage (24 hours’ holding time). The study included 20 crossbred rams (Pirot Pramenka x Wurttemberg x Ile de France), 12 months old. Animals were housed at the experimental sheep farm of the Institute for Animal Husbandry in Belgrade, Serbia. Semen was collected through electro ejaculation. The ejaculates were obtained from single services and routine field analysis of the semen was performed immediately after the collection. The semen was split and diluted with three extenders, namely Optidyl®, Andromed® and ultrahigh temperature processed (UHT) milk, in ratios of 1 : 50 or 1 : 100. The ejaculates were examined for sperm motility variables (sperm cell motility percentage, the progressive motility percentage, curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), sperm linearity (LIN), straightness (STR), amplitude of lateral sperm head displacement (ALH), beat cross frequency (BCF) and circular tracks), and sperm morphology (live sperm percentage, percentage of normal sperm forms with intact acrosome, percentage of abnormal sperm forms and total damaged acrosome) by computer-assisted sperm analysis (CASA) and classic sperm cytology after supravital eosin/nigrosine/trypan blue staining, respectively. It was observed that the type of extender used in diluting ram semen is an important factor in the successful short-term liquid preservation (at 4 °C) of ram spermatozoa. In conclusion, this study showed that egg yolk (Optidyl) and soybean (Andromed)-based extenders gave better results of both sperm morphology and sperm motility parameters compared with UHT milk.Keywords: Diluents, morphology, motility, sperm

17 Increased inbreeding levels negatively affect sperm kinetics and motility in Purebred Spanish horses

2021 ◽  
Vol 33 (2) ◽  
pp. 116
Author(s):  
Y. Pirosanto ◽  
A. Molina ◽  
M. Valera ◽  
J. Dorado ◽  
E. Terán ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Reproductive Traits ◽  
Study Groups ◽  
Inbreeding Coefficient ◽  
Computer Assisted ◽  
Effective Population ◽  
Mean Velocity ◽  
Sperm Analysis ◽  
Head Displacement

Reproductive performance is one of the key factors in livestock production. It is well known that reproductive traits are influenced by several genetic factors, such as the increase of individual inbreeding levels, which are associated with changes in sperm motility and shape in several species. In horses, the increase in inbreeding is a common problem because of the reduction in effective population size and the increase in selection intensity observed in several breeds. However, studies assessing the effect of high levels of inbreeding on the sperm quality of stallions are scarce. In the present study, we aimed to determine the effect of increased inbreeding levels and age on the sperm motility patterns of Purebred Spanish horses (PRE). We performed kinetic characterisation of 557 sperm samples of 82 PRE stallions aged between 3 and16 years, using computer-assisted sperm analysis (Androvision™, Minitube). We evaluated 5 parameters in 6 different fields per sample: curved line velocity (VCL, µm/s), velocity average path (VAP, µm/s), velocity straight line (VSL, µm/s), amplitude of lateral head displacement (ALH, µm), and beat-cross frequency (BCF, Hz). We determined the pedigree-based inbreeding coefficient (Fped) based on ∼300,000 PRE pedigree records to evaluate the inbreeding effect. Individuals were separated into 2 groups: highly inbred (n=339) and lowly inbred (n=218) according to an F value of 12.5%. Differences between groups were analysed using a generalized linear model. The analysis did not show significant differences (P>0.05) in the variables analysed with respect to the age of stallions. However, VAP, VCL, and AHL were lower in highly inbred than in lowly inbred animals (P<0.05), suggesting less velocity and amplitude of head displacement. In the case of BCF, no significant differences (P>0.05) were observed between the two study groups. In conclusion, age did not affect sperm quality parameters in the age group of stallions analysed. In addition, we demonstrated that high inbreeding coefficient reduced the mean velocity and trajectory pattern of spermatozoa in PRE.

scholarly journals Effects of chilled storage and pH of activating solution on different motility parameters in burbot (Lota lota) sperm

2018 ◽  
Vol 63 (No. 11) ◽  
pp. 429-434
Author(s):  
Zoltán Bokor ◽  
Balázs Csorbai ◽  
Levente Várkonyi ◽  
Zsolt Szári ◽  
Ferenc Fodor ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Chilled Storage ◽  
Straight Line ◽  
H 26 ◽  
Computer Assisted Sperm Analysis ◽  
Motility Parameters ◽  
Activating Solution

The effects of a simple saline solution prepared using two different pH (4.4 and 8.5) on sperm motility in burbot were investigated. Results were recorded during a 96-hour chilled storage (4°C) in 24-hour intervals. Measurements were focused on the detailed characteristics of motility using 12 parameters obtained from the Computer-assisted Sperm Analysis (CASA). Significantly higher progressive motility (pMOT), distance average path (DAP), distance curved line, distance straight line (DSL), average path velocity (VAP), curvilinear velocity, straight line velocity, and beat cross frequency (BCF) were observed with the activating solution buffered at pH 8.5 in comparison with pH 4.4. Already after 24 h a significant reduction was measured in pMOT (0 h: 49 ± 24%, 24 h: 12 ± 7%). Similar decreasing tendency was recorded only after 72 h in DAP (0 h: 26 ± 4 µm/s, 72 h: 19 ± 9 µm/s), DSL (0 h: 21 ± 5 µm/s, 72 h: 17 ± 8 µm/s), VAP (0 h: 59 ± 9 µm/s, 72 h: 43 ± 21 µm/s), and BCF (0 h: 28 ± 2 Hz, 72 h: 18 ± 10 Hz). The response of different investigated CASA parameters to different treatments varied in our experiments. According to our studies, numerous burbot sperm motility parameters are sensitive to chilled storage and to low pH of the activating solution. Our results could support the effective sperm quality assessment and successful artificial propagation process in burbot.

scholarly journals Metabolite Profiling of Pig Seminal Plasma Identifies Potential Biomarkers for Sperm Resilience to Liquid Preservation

2021 ◽  
Vol 9 ◽  
Author(s):  
Yentel Mateo-Otero ◽  
Pol Fernández-López ◽  
Jordi Ribas-Maynou ◽  
Jordi Roca ◽  
Jordi Miró ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Liquid Storage ◽  
Dependent Behavior ◽  
Downstream Gene Expression ◽  
Cell Signaling Pathways ◽  
C Nmr

Metabolomic approaches allow the study of downstream gene expression events since metabolites are considered as the products of cell signaling pathways. For this reason, many studies in humans have already been conducted to determine the influence of the metabolites present in seminal plasma (SP) on sperm physiology, and to identify putative biomarkers. However, in livestock species, these relationships are yet to be uncovered. Thus, the present study aimed to explore: (i) if concentrations of metabolites in pig SP are related to sperm quality and functionality, and (ii) if they could predict the sperm resilience to liquid storage at 17°C. To this end, 28 ejaculates were individually collected and split into three aliquots: one was used for SP analysis through nuclear magnetic resonance (NMR) spectroscopy; another served for the evaluation of sperm concentration and morphology; and the last one was utilized to determine sperm functionality parameters using computer-assisted sperm analysis (CASA) and flow cytometry after 0 h and 72 h of liquid-storage at 17°C. NMR analysis allowed the identification and quantification of 23 metabolites present in pig SP which, except for fumarate, were not observed to follow a breed-dependent behavior. Moreover, specific relationships between metabolites and sperm variables were identified: (i) glutamate, methanol, trimethylamine N-oxide, carnitine, and isoleucine were seen to be related to some sperm quality and functionality parameters evaluated immediately after semen collection; (ii) leucine, hypotaurine, carnitine and isoleucine were found to be associated to the sperm ability to withstand liquid storage; and (iii) Bayesian multiple regression models allowed the identification of metabolite patterns for specific sperm parameters at both 0 h and 72 h. The identification of these relationships opens up the possibility of further investigating these metabolites as potential sperm functional biomarkers.

16 COMPUTER-ASSISTED SPERM ANALYSIS SPERM PARAMETERS OF BOS TAURUS CRIOLLOS BULL SEMEN v. THOSE OF OTHER BOS TAURUS BREEDS IN ARGENTINA

2008 ◽  
Vol 20 (1) ◽  
pp. 88
Author(s):  
M. G. Lüssenhoff ◽  
G. Larraburu ◽  
R. Cavia ◽  
A. Garcia Guerra ◽  
G. M. Brogliatti
Keyword(s):  
Sperm Motility ◽  
Sperm Morphology ◽  
Bos Taurus ◽  
Hybrid Vigor ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Bull Semen ◽  
Significant Difference ◽  
Computer Assisted Sperm Analysis ◽  
Motility Parameters

Criollos is the oldest cattle breed in America and the world at large. The origin dates back to the first cattle brought by C. Columbus. They are well known for their toughness and longevity. Their genetic variability is another advantage to be taken into account in crossbreeding because it ensures high hybrid vigor. The objectives of the present study were to determine computer-assisted sperm analysis (CASA) motility parameters and sperm morphology of Criollos bull semen (brought from Parque Nacional Los Glaciares, Patagonia, Argentina) v. other Bos taurus semen. A total of 29 ejaculates of different adult bulls (Angus and Hereford) and 25 ejaculates of Criollos bulls were evaluated in an artificial insemination center between December 2004 and December 2005. Semen collection was done by electroejaculation (EE) in all breeds. Fresh semen was diluted in a semi-defined semen extender (Andromed; Minitüb, Tiefenbach, Germany) and frozen in an automatic freezer (Digicool, IMV, Paillette Crista, France). Stain morphology was done using eosin/nigrosin, and evaluated in an optic microscope with phase contrast. Parameters of volume, total concentration (CONC), motility %, motile progressive sperm %, velocity average path (VAP, μm s–1), velocity straight line (VSL, μm s–1), curvilinear velocity (VCL, μm s–1), and linearity (LIN, %) were determined by CASA (HTM-ceros 12.1, Berkley, CA, USA). The results obtained were analyzed statistically with a one-way ANOVA test and are summarized in Table 1. Results from the sperm motility analysis indicate that there were no significant differences (P > 0.05) in CONC, motility %, motile progressive %, or VSL between Criollos and other B. taurus bull semen. VAP and VCL rates were significantly higher in Criollos bull semen than in other B. taurus semen, and LIN was lower. Also, there was a significant difference in volume collected between both breeds. Regarding sperm morphology, Criollos bull semen is at the maximum limit acceptable for head defects (detached heads: 15%). These results suggest that Criollos bull semen is not different from other Bos taurus semen; however, it does show different velocity and linearity rates. Table 1. CASA sperm motility parameters for semen from Criollos v. other Bos taurus bulls This research was supported by Centro Genetico Bovino Eolia S.A.

Probiotic administration improves sperm quality in asthenozoospermic human donors

2017 ◽  
Vol 8 (2) ◽  
pp. 193-206 ◽  
Author(s):  
D.G. Valcarce ◽  
S. Genovés ◽  
M.F. Riesco ◽  
P. Martorell ◽  
M.P. Herráez ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Viability ◽  
Dna Fragmentation ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Bifidobacterium Longum ◽  
Fold Change ◽  
Sperm Chromatin ◽  
Computer Assisted ◽  
Sperm Analysis

The objective of this study is to analyse the effect of the ingestion of two selected antioxidant probiotics strains (Lactobacillus rhamnosus CECT8361 and Bifidobacterium longum CECT7347) on sperm quality parameters in asthenozoospermic males after three and six weeks of administration. Nine asthenozoospermic men without any medical treatment under similar diet conditions participated in the study. The quality of individual sperm samples was evaluated before (previous to ingestion), during (after 3 and 6 weeks of ingestion) and after probiotic administration (3 and 6 weeks after finishing the treatment). Sperm motility was evaluated by computer-assisted sperm analysis system, DNA fragmentation by sperm chromatin structure assay, cell viability by flow cytometry and measurement of intracellular H2O2 (reactive oxygen species; ROS) by flow cytometry using dichloro-dihydrofluorescein diacetate. Sperm motility was drastically improved after the treatment (approximately 6 fold change), DNA fragmentation was statistically reduced after probiotic administration from (approximately 1.2 fold change) and intracellular H2O2 level was decreased (approximately 3.5 fold change). Cell viability was not affected by the treatment. The significant improvement in sperm motility and the decrease in DNA fragmentation reported in this study provide preliminary evidence that probiotics could be administrated to improve motility and decrease DNA fragmentation and ROS levels in asthenozoospermic human males.

scholarly journals Quantitative sperm characteristics of Tankwa goats with special reference to hyperactivated motility

2021 ◽  
Vol 50 (5) ◽  
Author(s):  
Ngcauzele Ngcauzele ◽  
G. Van der Horst ◽  
A. Kotze ◽  
T. Jonker ◽  
L. Maree
Keyword(s):  
Sperm Motility ◽  
Animal Species ◽  
Sperm Quality ◽  
Semen Analysis ◽  
Procaine Hydrochloride ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Sperm Morphometry ◽  
Fertility Potential ◽  
Phosphate Buffered Saline

Computer-assisted semen analysis (CASA) is an automated and objective method of evaluating structural (e.g. morphology) and functional sperm parameters (e.g. motility and hyperactivation). Sperm hyperactivation is essential for successful fertilization and is thus an important aspect in determining the fertility potential of a male. In the current study, CASA was used for standard semen analysis and for comparison of the ability of phosphate buffered saline (PBS), BO sperm wash (10 mM caffeine), 4% lignocaine, and 5 mM procaine hydrochloride to induce hyperactivation in Tankwa goat spermatozoa. Twenty-nine ejaculates were collected from randomly selected male goats by electroejaculation. Although none of the four media affected percentage total sperm motility, lignocaine caused a significant decrease (P >0.05) in percentage progressive motility. Exposure to procaine resulted in an increase in swimming speed (P ≤0.05) and star-spin motility tracks, which are typical of sperm hyperactivation. Using PBS and procaine motility data from individually selected spermatozoa, receiver operator characteristic curves were constructed to distinguish the kinematic parameters employed as cut-off values for sperm hyperactivation. PBS and BO sperm wash did not induce hyperactivation (0.1 + 0.2% and 0.04  0.2% respectively), while lignocaine induced little hyperactivation (3.4 + 3.0%) and procaine hydrochloride had the highest percentage hyperactivation (25.3 + 13.6%). The large variation in hyperactivation (0–54.5%) may reflect inter-individual differences in sperm quality among these males. This study indicated procaine hydrochloride was the most promising hyperactivation-inducing medium for Tankwa goat spermatozoa and should be considered for similar assessments in other animal species Keywords: computer-aided sperm analysis, procaine hydrochloride, sperm kinematics, sperm morphometry, sperm motility

scholarly journals Post-Thaw Sperm Quality and Functionality in the Autochthonous Pig Breed Gochu Asturcelta

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1885
Author(s):  
José Néstor Caamaño ◽  
Carolina Tamargo ◽  
Inmaculada Parrilla ◽  
Felipe Martínez-Pastor ◽  
Lorena Padilla ◽  
...  
Keyword(s):  
Genetic Resource ◽  
Sperm Quality ◽  
Genetic Material ◽  
Mitochondrial Activity ◽  
Computer Assisted ◽  
Kinematic Parameters ◽  
Linear Mixed Effects ◽  
Sperm Analysis ◽  
Computer Assisted Sperm Analysis ◽  
Analysis System

Genetic resource banks (GRB) preserve the genetic material of endangered, valuable individuals or genetically relevant breeds. Semen cryopreservation is a crucial technique to reach these goals. Thus, we aimed to assess the sperm parameters of semen doses from the native pig breed Gochu Asturcelta stored at the GRB of Principado de Asturias (GRB-PA, Gijón, Spain), focusing on intrinsic and extrinsic (boar, season) factors. Two straws per boar (n = 18, 8–71 months of age) were thawed, pooled, and assessed after 30 and 150 min at 37 °C by CASA (computer-assisted sperm analysis system; motility and kinematic parameters) and flow cytometry (viability, acrosomal status, mitochondrial activity, apoptosis, reactive oxygen species, and chromatin status). The effects of age, incubation, and season on post-thawing quality were determined using linear mixed-effects models. Parameters were on the range for commercial boar breeds, with chromatin status (SCSA: fragmentation and immaturity) being excellent. Incubation decreased sperm quality and functionality. The boar age did not have a significant effect (p > 0.05), but the between-boar variability was significant (p < 0.001). The season significantly affected many parameters (motility, kinematics, viability, acrosomal status, mitochondrial activity), especially after 150 min of incubation. In general, samples collected in spring and summer showed higher quality post-thawing, the lowest in winter. In conclusion, the sperm doses from the Gochu Asturcelta breed stored at the GRB-PA showed excellent chromatin status and acceptable characteristics after thawing. Therefore, boar and seasonal variability in this autochthonous breed could be relevant for cryobank management.

Effect of transfection and co-incubation of bovine sperm with exogenous DNA on sperm quality and functional parameters for its use in sperm-mediated gene transfer

Zygote ◽  
2016 ◽  
Vol 25 (1) ◽  
pp. 85-97 ◽  
Author(s):  
María Elena Arias ◽  
Esther Sánchez-Villalba ◽  
Andrea Delgado ◽  
Ricardo Felmer
Keyword(s):  
Gene Transfer ◽  
Sperm Quality ◽  
Computer Assisted ◽  
Exogenous Dna ◽  
Sperm Analysis ◽  
Functional Parameters ◽  
Transgenic Embryos ◽  
Fertilization Capacity ◽  
Motility Parameters

SummarySperm-mediated gene transfer (SMGT) is based on the capacity of sperm to bind exogenous DNA and transfer it into the oocyte during fertilization. In bovines, the progress of this technology has been slow due to the poor reproducibility and efficiency of the production of transgenic embryos. The aim of the present study was to evaluate the effects of different sperm transfection systems on the quality and functional parameters of sperm. Additionally, the ability of sperm to bind and incorporate exogenous DNA was assessed. These analyses were carried out by flow cytometry and confocal fluorescence microscopy, and motility parameters were also evaluated by computer-assisted sperm analysis (CASA). Transfection was carried out using complexes of plasmid DNA with Lipofectamine, SuperFect and TurboFect for 0.5, 1, 2 or 4 h. The results showed that all of the transfection treatments promoted sperm binding and incorporation of exogenous DNA, similar to sperm incorporation of DNA alone, without affecting the viability. Nevertheless, the treatments and incubation times significantly affected the motility parameters, although no effect on the integrity of DNA or the levels of reactive oxygen species (ROS) was observed. Additionally, we observed that transfection using SuperFect and TurboFect negatively affected the acrosome integrity, and TurboFect affected the mitochondrial membrane potential of sperm. In conclusion, we demonstrated binding and incorporation of exogenous DNA by sperm after transfection and confirmed the capacity of sperm to spontaneously incorporate exogenous DNA. These findings will allow the establishment of the most appropriate method [intracytoplasmic sperm injection (ICSI) orin vitrofertilization (IVF)] of generating transgenic embryos via SMGT based on the fertilization capacity of transfected sperm.

242 EFFECTS OF TWO COMMERCIAL BOVINE SEMEN EXTENDERS FOR SHORT-TERM STORAGE ON MOTILITY PATTERN OF CHILLED BISON SEMEN

2011 ◽  
Vol 23 (1) ◽  
pp. 219
Author(s):  
B. M. Toosi ◽  
G. Gratton ◽  
C. Lessard ◽  
G. P. Adams
Keyword(s):  
Sperm Motility ◽  
Repeated Measures ◽  
Test Tube ◽  
Computer Assisted ◽  
San Jose ◽  
Short Term ◽  
Development Fund ◽  
Straight Line ◽  
Wood Bison

Difficulties of adequate cryopreservation of bison semen has limited the success of artificial insemination and in-vitro embryo production in bison. We evaluated the effects of short-term cooling on motility of bison sperm using two commercial semen extenders (Triladyl® and Andromed®; Minitube, Ingersoll, ON, Canada). Semen was collected by electroejaculation of mature wood bison (n = 3) and plains bison (n = 3) twice a week for 2 wk. Upon collection, the ejaculate was divided into 3 equal aliquots, which were then diluted 1:2 (vol/vol) in Triladyl or Andromed, or were not extended (n = 24 samples per treatment). Samples were maintained at 37°C until transfer to the laboratory (≤2 h). One millilitre of each sample was then placed into a test tube (15 mL, BD Falcon, BD Biosciences, San Jose, CA, USA) and were kept in water bath set at 5°C inside a walk-in refrigerator (4°C). Characteristics of sperm motility were evaluated before cooling (Day 0) and every 24 h after cooling for 5 days using a computer-assisted semen analyzer. Total motility (TM), progressive motility (PM), velocity curved line (VCL), velocity average path (VAP), and velocity straight line (VSL) were compared among treatments by ANOVA for repeated-measures. Values are expressed as mean ± SEM. After collection, the PM of the raw semen and semen extended in Triladyl or Andromed were not significantly different (63.1 ± 4.4%, 63.3 ± 3.1%, and 56.9 ± 4.5%, respectively). Cooling semen for a period of 24 h resulted in a decrease (P < 0.05) in PM in all 3 groups (4.4 ± 2.0%, 22.7 ± 2.9%, and 28.7 ± 4.3%, respectively). The PM of semen extended in Tryladyl or Andromed was greater than that of raw semen on Day 1 (P < 0.05). Semen extended in Triladyl and Andromed maintained PM on Day 2 (24.7 ± 3.3% and 21.8 ± 3.8%, respectively), but PM declined progressively to 1.1 ± 0.6% and 6.3 ± 2.1% by Day 5. A similar pattern was observed for the TM. The VCL, VAP, and VSL parameters for semen extended with Triladyl and Andromed decreased gradually between Day 0 and Day 5 (P < 0.05). From Day 1 to 4 after cooling, these velocity parameters were not significantly different between semen extended with Triladyl or Andromed; however, they were greater than those of raw semen (P < 0.05). All sperm velocity parameters for the raw semen declined by more than 60% between Days 0 and 2 (P < 0.05). On Day 0, VCL for semen extended with Andromed (152.2 ± 4.3) was greater than that of semen extended with Triladyl and raw semen (P < 0.05; 122.5 ± 7.0 and 122.4 ± 6.6, respectively). The VCL then decreased to 98.9 ± 12.9, 100.5 ± 10.8, and 18.6 ± 6.8 in Andromed, Triladyl and raw groups respectively on Day 2 (P < 0.05), followed by a further decline to 51.8 ± 14.3, 19.9 ± 10.0, and 9.0 ± 5.0, respectively, observed on Day 5. In conclusion, both Triladyl and Andromed improved characteristics of sperm motility of chilled bison semen. Despite an initial decrease within the first 24 h, bison sperm extended with Triladyl or Andromed maintained an acceptable degree of motility for up to 2 days after chilling to 5°C. Supported by Agriculture and Agri-Food Canada, Agriculture and Development Fund, and Canadian Animal Genetic Resources program.

23 Sperm quality of Pure Spanish stallions is affected by inbreeding coefficient and age

2020 ◽  
Vol 32 (2) ◽  
pp. 137
Author(s):  
Y. Pirosanto ◽  
M. Valera ◽  
A. Molina ◽  
J. Dorado ◽  
S. Demyda-Peyrás
Keyword(s):  
Sperm Motility ◽  
Inbreeding Depression ◽  
Negative Correlation ◽  
Semen Quality ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Inbreeding Coefficient ◽  
Semen Parameters ◽  
Computer Assisted

Inbreeding depression, a genetic condition produced by the mating of close-related individuals, has been associated with a reduction of fertility in several species. However, a loss in sperm quality was also associated with age. In horses, the few existing reports have described a tendency of both parameters to produce a negative effect on sperm quality. However, those reports were performed using a subjective evaluation of sperm motility. In the present study, a total of 692 ejaculates from 86 Pure Spanish stallions (PRE), aged between 3 and 22 years, were evaluated using a computer-assisted methodology to determine the effect of inbreeding in four semen parameters: free-gel volume (V), sperm concentration (C, by haemocytometer), and total (TM) and progressive (PM) sperm motility (by Spermvision sperm class analyser; Minitube). The inbreeding coefficient (F) was estimated using 300 000 PRE pedigree records approximately (minimum pedigree depth, eight equivalent complete generations; range, between 1 and 30.1%). Stallion, age, ejaculate, and season of semen collection were the variables included in the statistical model (general linear model), with ejaculate and season being the variables with a major effect (by variance components analysis). Our results showed that sperm concentration (r=−0.18; P&lt;0.0001) and volume (to a lesser extent) were reduced with advancing age, both showing a major decline after 15 years of age. To the contrary, sperm motility was not affected by age of the stallion. We also found a negative correlation between the inbreeding coefficient and ejaculate volume (r=−0.14; P&lt;0.001), with a marked decrease seen when F was between 7 and 20%. Also, a negative correlation was observed in PM (r=−0.08; P&lt;0.05), although to a lower extent. Conversely, C and TM were not affected by inbreeding depression (P&gt;0.05). In conclusion, our results demonstrated that high levels of inbreeding can compromise severely the sperm quality of the PRE stallion, which, subsequently, may have a negative influence on fertility. Ongoing studies using genomic data will help to detect genetic variants associated with stallion semen quality and how it is influenced by inbreeding in specific genomic regions.

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