Sorting and cryopreservation of goat sperm with or without phenolic compounds

ABSTRACT The objectives of this study were to evaluate goat sperm sorting in continuous Percoll® density gradients and gamete freezability, in the presence or absence of phenolic antioxidants. For this, semen pools were sorted, frozen, and evaluated. The non-selected group (NSg) presented lower progressive motility (PM), linearity (LIN), straightness (STR), and wobble (WOB) than the selected groups, and straight line velocity (VSL) compared to those with catechin or resveratrol. The amplitude of lateral head displacement (ALH) was higher in NSg, and quercetin reduced the mitochondrial membrane potential (MMP). After thawing, the NSg presented lower PM than the selected groups, VSL and VAP (average path velocity) than the selected group with or without catechin, LIN and WOB than the selected with or without catechin or resveratrol, and STR than the selected with catechin. Moreover, NSg presented higher ALH and BCF than the samples selected with or without catechin. Plasma membrane integrity and intact and living cells were higher in the selected groups, and MMP was lower in the NSg and the selected group with quercetin. Thus, centrifugation in Percoll® continuous density gradients is a viable methodology to select goat sperm compatible with the freezing, especially in the presence of catechin or resveratrol.

Download Full-text

Efecto de la composición racial sobre la calidad espermática de verracos

Agronomía Mesoamericana ◽

10.15517/ma.v29i3.32445 ◽

2018 ◽

Vol 29 (3) ◽

pp. 485 ◽

Cited By ~ 2

Author(s):

Anthony Valverde-Abarca ◽

Mónica Madrigal-Valverde ◽

Marlen Camacho-Calvo ◽

Adones Zambrana-Jiménez ◽

Leonardo López

Keyword(s):

Quality Evaluation ◽

Semen Quality ◽

Sperm Quality ◽

Active Movement ◽

Racial Groups ◽

Swine Farms ◽

Progressive Motility ◽

Head Displacement ◽

Straight Line ◽

Lateral Head

The assessment of the semen quality is an essential tool to improve the reproductive indexes in swine farms. The motility is the most important parameter that used in the sperm quality evaluation because it is associated with the energy of the spermatozoon. The aim of this research was to determine the influence of breed composition on reproductive variables of sperm quality, kinetics and semen motility in boars. During 2016, 240 ejaculates were collected from 63 boars with a mean age of 24.4±10.9 months. Six racial groups were identified: Duroc (D), Yorkshire (Y), Landrace (L), F1 Pietrain*Duroc (PD) and two genetic lines (LA and LB). The breeds with the highest ejaculate volume presented a higher total number of spermatozoa (P<0.05). The effect of the breed was significant (P<0.05) on sperm kinetics variables except for amplitude of lateral head displacement (ALH, μm). The Landrace breed presented the highest (P<0.05) percentage of static sperm (29.30±1.57). The most relevant differences (P<0.05) for total motility (MTOT) and progressive motility (MP), were presented between L and PD with values of 70.71±1.57; 77.48±1.09 and 51.80±1.97; 59.85±1.37% respectively. Adult boars (≥18 months) had higher volumes of ejaculate and total number of sperm than boars in the intermediate and young ages, however, for velocities (μm/s): curvilinear (VCL), straight line (VSL) and average path (VAP), adult boars only were different (P<0.05) of the boars in-between ages. Four sperm subpopulations (SP) were identified, SP1 (46.83%) characterized by a moderate speed, but of very progressive motility, SP2 (14.78%) with active movement, but not progressive, SP3 (8.45%) with low speeds and progressive path and SP4 (29.94%) with quick movement, but without progressivity.

Download Full-text

9 Single Layer Centrifugation of Bull Semen Through Percoll Plus® Before Cryopreservation

Reproduction Fertility and Development ◽

10.1071/rdv30n1ab9 ◽

2018 ◽

Vol 30 (1) ◽

pp. 144

Author(s):

A. Martins ◽

F. N. Marqui ◽

T. E. Cruz ◽

T. I. H. Berton ◽

D. G. Souza ◽

...

Keyword(s):

Semen Quality ◽

Single Layer ◽

Computer Assisted ◽

Treatment Groups ◽

Progressive Motility ◽

Bull Semen ◽

Head Displacement ◽

Straight Line ◽

Artificial Vagina ◽

Lateral Head

We previously reported that single layer centrifugation (SLC) with Percoll® (GE Healthcare, Uppsala, Sweden) of fresh bovine semen resulted in improved sperm progressive motility and movement, as evidenced by computer-assisted sperm analyzer (CASA) after freezing-thawing. However, no report has been found in the literature on the use of Percoll Plus® (PP; GE Healthcare), a nontoxic colloid, for the same purpose. Thus, this study aimed to verify the effects of SLC-PP before bull sperm freezing on sperm kinematics after cryopreservation. Ejaculates were collected from 3 Nellore bulls (6 from each) using an artificial vagina. After collection, the semen was assessed and pooled, and then 1 billion spermatozoa either diluted [D; 1:2 (v/v)] in freezing extender (FE, without glycerol) or undiluted (UD) was layered on top of a 9-mL column of PP (in 15-mL centrifuge tubes) at concentrations of 70% or 90% to form the 70D, 70UD, 90D, and 90UD treatment groups. Following centrifugation for 13 min at 839 × g [except for the control (C) group], the supernatant was removed and the sperm pellet diluted to 50 × 106 sperm mL−1 in FE medium plus glycerol. Then, frozen–thawed sperm samples were analysed by CASA (MMC Sperm, St. Petersburg, Russia) for the following parameters: total motility (TM, %), progressive motility (PM, %), curvilinear velocity (VCL, µm−1), straight line velocity (VSL, µm s−1), average path velocity (VAP, µm s−1), amplitude of lateral head displacement (ALH, µm), beat cross frequency (BCF, Hz), linearity (LIN, %), and straightness (STR, %). For statistical analyses, ANOVA and Student-Newman-Keuls test were used. Data are presented as mean ± SEM with P < 0.05 taken as significant. No difference was found among the groups for TM, VSL, BCF, and STR. However, the percentage of PM was higher (P < 0.05) in the SLC-selected sperm samples (values ranging from 42.0 ± 7.0 to 47.4 ± 11.4) than in C (28.8 ± 5.0), and ALH was lower in 70UD (1.6 ± 0.12) and 70D (1.7 ± 0.10) than in C (1.9 ± 0.2). Moreover, 70UD (49.0 ± 1.0), 90UD (50.0 ± 3.0), and 90D (50.0 ± 4.0) displayed higher percentage of LIN (P < 0.05) compared with C (45.0 ± 2.0) and 70D (48.0 ± 3.0). On the other hand, similar results were obtained for VCL (from 126.3 ± 8.0 to 130.0 ± 20.5) and VAP (from 82.7 ± 14.5 to 85.1 ± 6.9) in C, 70UD, and 70D, but these values differed (P < 0.05) from those for VCL in 90UD (104.6 ± 10.3) and 90D (97.2 ± 22.0) as well as for VAP in 90UD (72.2 ± 11.0) and 90D (71.8 ± 9.6). These are the first data demonstrating favourable influences of SLC with 70% Percoll Plus® to select distinct sperm subpopulations as evidenced by enhanced PM, LIN, and ALH. Thus, SLC-PP could optimize the production of frozen bull semen by decreasing the number of sperm per insemination dose, and help to circumvent limitations associated with the poor semen quality sometimes found in bulls of high genetic merit. This research was funded by FAPESP # 2015/20986-3, MasterFertility and Tairana Artificial Insemination Station, Brazil.

Download Full-text

Computer-Assisted Sperm Analysis of Freezable and Nonfreezable Mithun (Bos frontalis) Semen

Journal of Animals ◽

10.1155/2014/675031 ◽

2014 ◽

Vol 2014 ◽

pp. 1-6 ◽

Cited By ~ 3

Author(s):

P. Perumal ◽

S. K. Srivastava ◽

S. K. Ghosh ◽

K. K. Baruah

Keyword(s):

Objective Analysis ◽

Computer Assisted ◽

Sperm Analysis ◽

Progressive Motility ◽

Head Displacement ◽

Straight Line ◽

Computer Assisted Sperm Analysis ◽

Lateral Head ◽

Motility Parameters

The present study was undertaken to assess the motility and velocity parameters of sperm of freezable and nonfreezable ejaculates by computer-assisted sperm analyser (CASA) such as Hamilton-Thorne Semen Analyser IVOS 11 in mithun semen. Fifty ejaculates (twenty-five ejaculates each for freezable and nonfreezable semen ejaculates) were collected from ten matured mithun bulls. CASA parameters, motility parameters such as forward progressive motility (FPM) (%), nonprogressive motility (NPM) (%), total motility (TM) (%), and static sperms (SM) (%); velocity parameters such as curvilinear velocity (VCL) (μm/sec), straight line velocity (VSL) (μm/sec), average path velocity (VAP) (μm/sec), linearity (LIN) (%), straightness (STR) (%), wobble (WOB) (%), amplitude of lateral head displacement (ALH) (μm), and beat/cross-frequency (BCF) (Hz) were measured by CASA analyser. The result revealed that these parameters varied significantly (P<0.05) between the freezable and nonfreezable ejaculates and freezable ejaculates have significantly (P<0.05) higher value than nonfreezable ejaculates. It was concluded that most of the CASA parameters were significantly lower in nonfreezable ejaculates than in freezable ejaculates in mithun and confirmed that the CASA was effective for a quick and objective analysis of motility and velocity parameters in mithun semen.

Download Full-text

501. BACTERIOSPERMIA AND ITS CONTROL IN BUBALINE SEMEN

Reproduction Fertility and Development ◽

10.1071/srb09abs501 ◽

2009 ◽

Vol 21 (9) ◽

pp. 102

Author(s):

S. M. H. Andrabi ◽

M. Shahab

Keyword(s):

Bacterial Species ◽

Membrane Integrity ◽

Negative Control ◽

Computer Assisted ◽

Fertility Rates ◽

Head Displacement ◽

Straight Line ◽

Frozen Semen ◽

Lateral Head

The present study was designed to investigate the bacterial species incriminated in bubaline semen and to find out the effectiveness of antibiotics (GTLS; gentamycin, tylosin and linco-spectin or SP; streptomycin and penicillin) in cryodiluent on bacterial control and quality of buffalo bull spermatozoa. For this purpose four experiments were conducted. In experiment 1, a total of 11 bacterial species were identified from buffalo ejaculates. The predominant bacteria were Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa in the ejaculates. In experiment 2, total aerobic bacterial counts in post-thaw samples were lower (P<0.05) in GTLS than in SP or NC (negative control). Fewer bacterial genera were identified in semen samples having GTLS than SP. Majority of the bacterial isolates from ejaculates showed more sensitivity towards GTLS than SP. In experiment 3, motilities (visual and computer-assisted), velocities (straight-line, average path and curvilinear), amplitude of lateral head displacement and plasma membrane integrity in post-thaw semen samples did not differ (P>0.05) due to antibiotics. Spermatozoal abnormalities (acrosome, head, mid-piece and tail) were lower (P<0.05) in GTLS and SP than in NC. In experiment 4, the fertility rates for SP-based vs. GTLS-containing frozen semen of buffalo bull were 42.8 and 55.2%, respectively. The results for GTLS were significantly higher than SP. The fertility rates also differed significantly in the first and second batch of inseminations performed with SP or GTLS-based cryopreserved semen of buffalo bull. In conclusion, a number of bacterial species are isolated from bubaline semen. Bacterial and seminal quality measured by standard laboratory tests and field fertility trials indicate that GTLS is more suitable in extender for cryopreservation of buffalo bull spermatozoa.

Download Full-text

Vitamin C ameliorates tetrahydrocannabinol-induced spermatotoxicity in-vitro

10.21203/rs.3.rs-34960/v3 ◽

2020 ◽

Author(s):

Abdullateef Isiaka Alagbonsi ◽

Luqman Aribidesi Olayaki

Keyword(s):

Vitamin C ◽

Cannabinoid Receptors ◽

Group Iv ◽

Group Vi ◽

Spermatozoa Motility ◽

Progressive Motility ◽

Straight Line ◽

Lateral Head

Abstract Background: We investigated the in-vitro effects of vitamin C on delta-9-tetrahydrocannabinol (THC) -induced reduction in spermatozoa motility and kinematics. Methods: Six rats were used for the study. Semen from each of the 6 rats was randomly divided into 6 groups such that each rat’s semen was in all of the groups. Groups I-III received placebo, THC (1 mM), and vitamin C (5 mM) respectively. Group IV was pretreated with cannabinoid receptors’ blockers (CBs-) 1 and 2, followed by THC. Groups V and VI received THC and vitamin C, but group VI was additionally pre-treated with CBs-. Results: The spermatozoa progressive motility, average path velocity (VAP), curvilinear velocity (VCL), straight-line velocity (VSL), amplitude of lateral head (ALH) and beat cross frequency (BCF) were reduced by THC (6.08±1.16%; 5.64±0.82 µm/s; 6.96±0.74 µm/s; 2.75±0.23 µm/s; 0.31±0.02 µm; and 0.78±0.08 Hz respectively) but increased by vitamin C (51.20±1.32 %; 17.90±0.21 µm/s; 25.11±0.96 µm/s; 8.80±0.27 µm/s; 0.75±0.01 µm; and 3.15±0.03 Hz respectively) when compared to control (39.72±0.38 %; 13.70±0.29 µm/s; 18.04±0.58 µm/s; 7.54±0.34 µm/s; 0.65±0.02 µm; and 2.79±0.01 Hz respectively). Vitamin C inhibited the THC-induced reduction in these parameters (37.36±0.73 %; 10.98±0.45 µm/s; 13.58±0.30 µm/s; 7.11±0.22 µm/s; 0.58±0.01 µm; and 2.60±0.01 Hz respectively) in the absence of CBs- 1 and 2, and even caused additional increases in progressive motility (49.54±1.01 %), VAP (15.70±0.38 µm/s) and VCL (22.53±0.29 µm/s) above the control levels with CBs-.Conclusion: Vitamin C ameliorates the THC-induced reduction in spermatozoa motility in-vitro by modulation of their kinematics.

Download Full-text

90 EFFECT OF EGG YOLK ON THE KINEMATICS AND ACROSOME MEMBRANE INTEGRITY OF COOLED-REWARMED CANINE SPERMATOZOA

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab90 ◽

2010 ◽

Vol 22 (1) ◽

pp. 204

Author(s):

J. Dorado ◽

M. J. Galvez ◽

M. R. Murabito ◽

S. Demyda ◽

L. J. De Luca ◽

...

Keyword(s):

Semen Analysis ◽

Membrane Integrity ◽

Egg Yolk ◽

Computer Assisted ◽

Mean Values ◽

Head Displacement ◽

Healthy Dogs ◽

Acrosome Integrity ◽

Digital Manipulation ◽

Lateral Head

Tris-egg yolk-based diluents provide adequate cryoprotection for the sperm of most species. This study was conducted to compare the ability of Tris-glucose extender containing 2 different concentrations of egg yolk to maintain sperm motility and acrosome integrity of canine spermatozoa during 72 h of preservation. For this purpose, a total of 20 ejaculates from 4 clinically healthy dogs (2 Spanish Greyhound, 1 German Pointer, and 1 Crossbreed) were collected by digital manipulation. The sperm-rich fraction of each ejaculate was divided into 2 aliquots. Then, they were diluted in Tris-based extender and centrifuged at 700g for 8 min. Sperm pellets were resuspended in either Tris buffer added to 20% (EY20) or 10% centrifuged egg yolk (EY10) and cooled to 5°C over 72 h. The effects of these extenders on motility and acrosome integrity were assessed objectively using a computer-aided semen analyzer (Sperm Class Analyzer, Microptic SL, Spain) and Spermac® staining, respectively. Each cooled-rewarmed semen sample was evaluated after 24, 48, and 72 h of preservation. Sperm motion parameters shown by computer-assisted semen analysis (CASA) are progressively motile (PMS) and motile spermatozoa (MS), curvilinear velocity (CLV), average path velocity (APV), progressive speed (SLV), and lateral head displacement (LHD). Data were statistically analysed by ANOVA. Dependent variables expressed as percentages were arsine-transformed before analysis. Differences between mean values were evaluated by the Duncan method. Data were presented as mean ± SEM. Differences were considered significant when P < 0.05. Analyses were performed using the statistical package SPSS 12.0. A total of 98 172 motile sperm trajectories were analyzed by CASA: 52 259 in EY20 and 45 913 in EY10. After 24, 48, and 72 h of preservation, MS and PMS were statistically higher (P < 0.01) in EY20. No significant differences were found for LHD using either extender over a 72-h period. No significant differences were observed for CLV using either extender during the first 2 days. At Day 3, CLV data were significantly higher (P < 0.01) in EY20. Similarly, from Day 2, APV was significantly higher (P < 0.001) in EY20. After 24 h of preservation, SLV was statistically higher (P < 0.001) in EY10, whereas the opposite tendency was found at Day 3. No significant differences were observed for SLV using either extender after 48 h of preservation. During the first 2 days, acrosome integrity was statistically higher (P < 0.001) in EY20. At hour 72, higher acrosome integrity (P < 0.001) was observed in EY10. In conclusion, we have observed that the EY20 extender provided higher motility after 72 h of chilled preservation; however, the acrosome membrane integrity was better preserved in EY10.

Download Full-text

Vitamin C Ameliorates Tetrahydrocannabinol-Induced Spermatotoxicity In-Vitro

10.21203/rs.3.rs-34960/v1 ◽

2020 ◽

Author(s):

Abdullateef Isiaka Alagbonsi ◽

Luqman Aribidesi Olayaki

Keyword(s):

Vitamin C ◽

Sperm Motility ◽

Cannabinoid Receptors ◽

Group V ◽

Progressive Motility ◽

Straight Line ◽

Lateral Head

Abstract Background: We investigated the in-vitro effects of vitamin C on delta-9-tetrahydrocannabinol (THC)-induced reduction in sperm motility and kinematics. Methods: Rats semen were randomly divided into 5 groups. Groups I-III received placebo, THC (1 mM), and vitamin C (5 mM) respectively. Groups IV and V received THC and vitamin C, but group V was additionally pre-treated with cannabinoid receptors’ blockers (CBs-) 1 (SR141716) and 2 (AM-630). Results: The sperm progressive motility, average path velocity (VAP), curvilinear velocity (VCL), straight-line velocity (VSL), amplitude of lateral head (ALH) and beat cross frequency (BCF) were reduced by THC but increased by vitamin C when compared to control. Vitamin C inhibited the THC-induced reduction in these parameters in the absence of CBs- 1 and 2, and even caused additional increases in progressive motility, VAP and VCL above the control levels with CBs-. Conclusion: In conclusion, vitamin C ameliorates THC-induced reduction in sperm motility in-vitro by modulation of sperm kinematics.

Download Full-text

Effect of three commercial extenders on sperm motility and fertility in liquid ram semen stored at 15 °C or 5 °C

Acta Veterinaria Hungarica ◽

10.1556/004.2019.043 ◽

2019 ◽

Vol 67 (3) ◽

pp. 430-444

Author(s):

Ander Arando ◽

Juan Vicente Delgado ◽

José Manuel León ◽

Sergio Nogales ◽

Francisco Javier Navas-González ◽

...

Keyword(s):

Sperm Motility ◽

Soybean Lecithin ◽

Egg Yolk ◽

Kinematic Parameters ◽

Progressive Motility ◽

Head Displacement ◽

Liquid Storage ◽

Lateral Head

The effect of different extenders on sperm motility and fertility was evaluated during liquid storage of ram semen at 5 °C and 15 °C. The semen was collected, pooled and diluted in three commercial extenders: Inra 96® (INRA) based on skimmed milk, Biladyl® A fraction (BIL) based on egg yolk, and Ovixcell® (OVIX) based on soybean lecithin. Then, sperm motility was evaluated at 0, 6, 24, 48, 72 and 96 h. In order to evaluate fertility, samples stored at 15 °C were used after dilution in INRA and OVIX. Results showed that progressive motility was significantly higher up to 72 h of storage in sperm samples maintained at 5 °C in comparison with 15 °C, similarly for each tested diluent. When samples were stored at 5 °C in OVIX, kinematic parameters such as velocity (except curvilinear velocity, VCL), trajectory [linearity (LIN), straightness (STR), wobble (WOB)], amplitude of lateral head displacement (ALH) and beat/cross frequency (BCF) were higher than in INRA and BIL. No significant differences in pregnancy rate were detected between INRA (62.6%) and OVIX (58.9%). In conclusion, liquid storage at 5 °C with OVIX extender is an interesting option since non-animal components are used, and this extender offers similar in vitro and in vivo efficacy as other extenders containing animal components.

Download Full-text

72 EFFECT OF SINGLE-LAYER CENTRIFUGATION WITH EQUIPURE™ ON MOTILITY KINEMATICS OF FROZEN - THAWED DONKEY SPERM

Reproduction Fertility and Development ◽

10.1071/rdv25n1ab72 ◽

2013 ◽

Vol 25 (1) ◽

pp. 183 ◽

Cited By ~ 1

Author(s):

I. Ortiz ◽

J. Dorado ◽

D. Acha ◽

L. Ramirez ◽

M. Urbano ◽

...

Keyword(s):

Sperm Quality ◽

Single Layer ◽

Bottom Layer ◽

Computer Assisted ◽

Kinematic Parameters ◽

Sperm Analysis ◽

Progressive Motility ◽

Head Displacement ◽

Straight Line ◽

Computer Assisted Sperm Analysis

Single-layer centrifugation (SLC) with EquipureTM Bottom Layer has been used to enhance the quality of stallion semen samples; however, no studies have been performed on donkeys. The aim of this study was to determine if SLC with EquipureTM Bottom Layer improves kinematic parameters on frozen–thawed donkey sperm. Semen was collected from 4 Andalusian donkeys by artificial vagina. Three ejaculates from each donkey were centrifuged with EquiproTM, supernatant was removed, and pellet was re-extended in the freezing medium GentTM to a final concentration of 200 × 106 spermatozoa per milliliter. Sperm were slowly cooled to 5°C for 2 h, loaded in 0.5-mL plastic straws, and frozen in liquid-nitrogen vapors. After at least one week of storage, straws were thawed in a water bath at 37°C for 30 s. After thawing, semen samples were divided in 2 aliquots: aliquot 1 was used as such (control) and aliquot 2 was processed by SLC using EquipureTM Bottom Layer. Computer-assisted sperm analysis was performed, and sperm kinematics total motility (%), progressive motility (%), curvilinear velocity (VCL; µm s–1), velocity straight line (VSL; µm s–1), velocity average path (VAP; µm s–1), linearity (LIN; %), straightness (STR; %), wobble (WOB; %), lateral head displacement (ALH; µm), and beat cross frequency (BCF; Hz) were statistically compared using GLM model between frozen–thawed semen samples processed or not with EquipureTM. Results were expressed as mean ± standard error. Significant differences (P < 0.05) were found between SLC-selected and unselected semen for total motility (77.44 ± 5.83 v. 58.89 ± 6.07), progressive motility (76.88 ± 4.52 v. 56.59 ± 5.44), VCL (137.50 ± 0.75 v. 133.0 ± 0.99), LIN (69.43 ± 0.31 v. 68.23 ± 0.41), STR (78.45 ± 0.29 v. 76.90 ± 0.37), WOB (85.06 ± 0.18 v. 83.91 ± 0.26), ALH (2.76 ± 0.01 v. 2.44 ± 0.01), and BCF (9.13 ± 0.05 v. 8.53 ± 0.06), respectively. No significant differences were observed for VSL (102.89 ± 0.70 v. 104.32 ± 0.95) and VAP (123.21 ± 0.71 v. 121.50 ± 0.98). Most of the computer-assisted sperm analysis parameters used in the present study have been previously identified as reliable markers of sperm motility in relation to sperm quality and fertility. It has also been reported that VCL appears to be critical for the formation of the sperm reservoir and penetration of the zona pellucida. In addition, other variables improved in the SLC-selected samples have been described as measure of progressivity (LIN, STR) and spermatozoa vigor (BCF, ALH). These preliminary results suggest an additional option for improving sperm quality in donkey semen doses. In conclusion, SLC with EquipureTM can be used to enhance kinematic parameters on frozen–thawed donkey sperm.

Download Full-text

Characterisation of movement pattern and velocities of stallion spermatozoa depending on donor, season and cryopreservation

Acta Veterinaria Hungarica ◽

10.1556/avet.51.2003.3.13 ◽

2003 ◽

Vol 51 (3) ◽

pp. 395-408 ◽

Cited By ~ 6

Author(s):

Christina Warnke ◽

A. Tuchscherer ◽

Hannelore Alm ◽

W. Kanitz ◽

S. Blottner ◽

...

Keyword(s):

Breeding Season ◽

Rank Order ◽

Movement Pattern ◽

Mean Velocity ◽

Sperm Analysis ◽

Head Displacement ◽

Straight Line ◽

Highly Correlated ◽

Casa System ◽

Lateral Head

The aim of the study was to compare different types of movement pattern and velocities of stallion spermatozoa depending on cryopreservation during breeding and non-breeding season. Ejaculates were collected from four stallions during May (n = 24) and December (n = 24). Parameters of sperm movement were evaluated by computer-aided sperm analysis (CASA) system, and included percentages of motile spermatozoa, different patterns of motility, the velocity, linearity (LIN), amplitude of lateral head displacement (ALH) and beat-cross frequency (BCF). In winter the average percentages of motility were slightly higher compared to the breeding season in May (70.8 ± 12.7% vs. 66.8 ± 12.2%, respectively). Cryopreservation and thawing led to a significant decrease in the number of motile sperm to 11.3 ± 5.8% in May and 15.6 ± 7.0% in December. The pattern of motility was also changed. Detailed analysis by CASA demonstrated that cryopreservation resulted in a shift from the proportions of linear to more non-linear motile spermatozoa and to a significant increase of local motile and hyperactivated spermatozoa. Mean velocity of fresh motile spermatozoa differed between May and December (119.1 ± 43.9 vs. 164.4 ± 66.4 µm/sec, respectively; P < 0.05). Cryopreservation and thawing led to a slight increase of curvilinear velocity (VCL) and straight line velocity (VSL). The motility analysis has shown that the parameters BCF and ALH were highly correlated in stallion spermatozoa (r = -0.67; P < 0.001). The BCF of stallion spermatozoa was slightly reduced in the non-breeding season. Altogether, the influence of factors on the motility of stallion spermatozoa has the following rank order: cryopreservation (P < 0.0001) ≯ stallion (P < 0.001) ≯ season (P < 0.05).

Download Full-text