P–126 A comparison of the efficacy of sperm freezing using high security tubes versus high security straws

Abstract Study question Are there any advantages in using High security tubes rather than High Security straws for conventional slow sperm freezing? Summary answer Freezing sperm in High Security tubes (HST) improved post-thaw recovery rate and motility, and also reduced processing and handling compared to High Security straws (HSS). What is known already The use of High Security freezing consumables (HSFC) in an IVF setting is a safe and effective way of eliminating concerns related to viral cross-contamination during storage. The lower diameter of HSS does make them susceptible to warming during handling. The HSFC used in this study is the only CE marked products that are made of resin, leak-proof and shatter-proof in all cryogenic temperatures even in LN2. No previous studies have compared the use of HST with HSS for conventional human sperm freezing. This study sets out to investigate the performance of HST compared to HSS. Study design, size, duration The study was designed as a controlled split-sample study with blind post-thaw analysis. Following the routine WHO analysis of 20 semen samples, the remainder of each of the samples was evenly divided and cryopreserved by conventional slow freezing in each of the two different HSFC. The freeze was conducted simultaneously by the same practitioner, employing the same freezing protocol and cryoprotectant. The pre-freeze and post-thaw concentration, total and progressive sperm motility were recorded. Participants/materials, setting, methods At one IVF clinic, semen samples with sperm density ≥15million/ml, ≥40% motility, ≥1.5ml were included. Cryoprotectant (SpermFreeze, Fertipro) was added dropwise to unprepared semen and kept at room temperature for 10 minutes before loading into HSFC (0.5ml CBS™HSS; CBS™HST). HSFC were heat-sealed (SYMS; SYMSIII sealers) and placed in vapour for 30 minutes before plunging into LN2. Samples were thawed by immersion in a 37Cº water bath for 5 minutes and analysed using WHO methods. Main results and the role of chance Paired-t test was used to compare the percentage motility between the different HSFC. All analysis was considered statistically significant when p < 0.01. We demonstrated that the sperm recovery rate (Percentage total motility post-thaw/ Percentage total motility pre-freeze) in HST was 66.63 ± 14.94 (mean ± standard deviation) compared to 40.80 ± 14.69 in HSS. In the HSS, the percentage post-thaw total motility was 19.99 ± 7.21 and the percentage post-thaw progressive motility was 12.26 ± 2.59. In the HST, the percentage post-thaw total motility was 32.57 ± 8.33 and the percentage post-thaw progressive motility was 23.08 ± 5.53. The overall improvement when using HST against HSS was 12.53 ± 5.69, 10.44 ± 5.29 for the total motility and the progressive motility respectively. Comments were recorded regarding the handling and the condition of the HSS and HST for each freeze event. Neither device displayed any leakage of LN2 or any explosion during the warming. The freezing process was easier and faster using HST rather than HSS. It was also noted that the entire sample can be recovered from the HST, unlike the HSS. Limitations, reasons for caution The study looked at sperm recovery in terms of motility only. DNA damage was not considered as a parameter of sperm quality. Also, fertilization, pregnancy rates, live birth rates and the use of poorer quality sperm samples have not been investigated. Wider implications of the findings: For conventional sperm freezing, the use of HST resulted in improved sperm motility and progression post-thaw, when compared to HSS. This finding supports the use of HST to improve the post thaw quality of sperm, benefitting patients with own frozen samples, recipients of donor sperm and donor sperm banks. Trial registration number Not applicable.

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85 THE COMPARISON OF SPERM FREEZABILITY USING TWO EGG YOLK-FREE DILUENTS IN ZANDI RAM

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab85 ◽

2010 ◽

Vol 22 (1) ◽

pp. 201

Author(s):

R. Ardebili ◽

A. Towhidi ◽

S. Zeinodini ◽

M. Bahreini ◽

E. Dirandeh

Keyword(s):

Least Squares ◽

Liquid Nitrogen ◽

Sperm Motility ◽

Recovery Rate ◽

Sperm Quality ◽

Egg Yolk ◽

Freezing And Thawing ◽

Automatic Equipment ◽

Progressive Motility ◽

Motility Of Sperm

The aim of this study was to investigate the effect of 2 different commercial egg yolk-free extenders, Bioxcell and AndroMed, on post-thaw ram sperm motility and recovery rate. A total of 10 ejaculates were collected from 5 adult Iranian Zandi rams using artificial vagina during 3 weeks, once per week. Preexamination was conducted on semen, and semen motility >70% were selected. Semen samples were pooled and diluted with Bioxcell or AndroMed. Pooled semen was split into 2 treatments and extended to a final concentration of 200 × 108 spermatozoa per mL with extenders. The extended semen was manually packed into 0.25-mL mini-straws and sealed with PVC powder. After 4 h of equilibration in a waterbath at 5°C, the straws were dried with paper tissue, placed horizontally on a rack, and transferred to an isothermal box to be frozen in vapor 5 cm above liquid nitrogen. After 10 min, the straws were plunged into liquid nitrogen. Freezing was conducted using IMV semi-automatic equipment. The percentage of motility and progressive motility of sperm were evaluated before freezing (at 37°C), after refrigeration (at 5°C) and thawing, and recovery rate was calculated. To evaluate indices of sperm motility including the percentage of motile sperm and the progressive forward motility, semen was diluted with 0.9% NaCl w/v (1:100). Ten μL of diluted semen was placed on the prewarmed (37°C) microscope slide (76.2 mm × 24.5 mm; Pearl, China) covered with a cover slip (24 × 24 mm; Menzel-Glaser, Germany) and examined 10 different fields under a phase contrast microscope (Nikon, Tokyo, Japan), at ×200. Dilution factor 1:5 was used. The microscope was equipped with warm plate set at 37°C. Data were analyzed using Proc GLM of SAS. The effect of extender on sperm qualitative characteristics after thawing was significant. Least squares mean percent motility and progressive motility of AndroMed (36.05% ± 0.75; 29.0% ± 0.84) was significantly higher (P < 0.01) than Bioxcell (8.02% ± 0.35; 4.0% ± 0.60); least squares mean recovery rate in AndroMed (34.23% ± 1.05) also was higher (P < 0.01) than Bioxcell (10.0% ± 0.92). Results suggested that AndroMed in comparison with Bioxcell had more ability to preserve sperm quality during the freezing and thawing process.

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117. THE COMPARISON OF SPERM FREEZABILITY USING TWO EGG YOLK-FREE DILUENTS IN ZANDI RAM

Reproduction Fertility and Development ◽

10.1071/srb09abs117 ◽

2009 ◽

Vol 21 (9) ◽

pp. 36

Author(s):

R. Ardebili ◽

A. Towhidi ◽

S. Zeinodini ◽

M. Bahreini

Keyword(s):

Recovery Rate ◽

Sperm Quality ◽

Calculated Data ◽

Egg Yolk ◽

Freezing Process ◽

Automatic Equipment ◽

Progressive Motility ◽

Motility Of Sperm ◽

Qualitative Characteristics

The aim of this study was to investigate the effect of two kinds of commercial egg yolk-free extenders: Bioxcell and AndroMed on in vitro sperm function of sheep. A total of 10 ejaculates were collected from 5 adult Iranian Zandi rams using artificial vagina. Semen samples were mixed and diluted with Bioxcell or AndroMed. Freezing was conducted using imv semi-automatic equipment. The percentage of motility and progressive motility of sperm were evaluated before freezing (at 37°C), after refrigeration (at 5°C) and thawing, and recovery rate was calculated. Data was analyzed using proc GLM of SAS. The effect of extender on sperm qualitative characteristics after thawing was significant. Mean percentage of motility, progressive motility and recovery rate were higher (P≤0.01) in AndroMed than Bioxcell. Results suggested that AndroMed in comparison with Bioxcell had more ability to preserve sperm quality during freezing process.

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The Hydrogen Storage Nanomaterial MgH2 Improves Irradiation-induced Male Fertility Impairment by Suppressing Oxidative Stress

10.21203/rs.3.rs-1207725/v1 ◽

2022 ◽

Author(s):

Jing Ma ◽

Suhe Dong ◽

Hongtao Lu ◽

Zhongmin Chen ◽

Huijie Yu ◽

...

Keyword(s):

Particle Size ◽

Hydrogen Storage ◽

Sperm Motility ◽

Male Fertility ◽

Sperm Quality ◽

Hormone Secretion ◽

Phase Cell ◽

Sperm Density ◽

Radiation Induced

Abstract ObjectiveThis study aimed to reveal the protective effect of hydrogen storage nanomaterial MgH2 on radiation-induced male fertility impairment.MethodsThe characterization of MgH2 were analyzed by scanning electron microscopy (SEM) and particle size analyzer. The safety of MgH2 were evaluated in vivo and in vitro. The radioprotective effect of MgH2 on the reproductive system were analyzed in mice, including sperm quality, genetic effect, spermatogenesis, and hormone secretion. ESR, flow cytometry and western blotting assay were used to reveal the underlying mechanisms.ResultsMgH2 had an irregular spherical morphology and a particle size of approximately 463.2 nm, and the content of Mg reached 71.46%. MgH2 was safe and nontoxic in mice and cells. After irradiation, MgH2 treatment significantly protected testicular structure, increased sperm density, improved sperm motility, reduced deformity rates, and reduced the genetic toxicity. Particularly, the sperm motility were consistent with those in MH mice and human semen samples. Furthermore, MgH2 treatment could maintain hormone secretion and testicular spermatogenesis, especially the generation of Sertoli cells, spermatogonia and round sperm cells. In vitro, MgH2 eliminated the [·OH], suppressed the irradiation-induced increase in ROS production, and effectively alleviated the increase in MDA contents. Moreover, MgH2 significantly ameliorated apoptosis in testes and cells and reversed the G2/M phase cell cycle arrest induced by irradiation. In addition, MgH2 inhibited the activation of radiation-induced inflammation and pyroptosis.ConclusionMgH2 improved irradiation-induced male fertility impairment by eliminating hydroxyl free radicals.

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121 Effect of pentoxifylline on motility of good- and poor-quality frozen-thawed equine sperm

Reproduction Fertility and Development ◽

10.1071/rdv32n2ab121 ◽

2020 ◽

Vol 32 (2) ◽

pp. 187

Author(s):

M. Felix ◽

I. Ortiz ◽

H. Resende ◽

J. Brom-de-Luna ◽

C. Love ◽

...

Keyword(s):

Sperm Motility ◽

Sperm Quality ◽

Phosphodiesterase Inhibitor ◽

Petri Dish ◽

Poor Quality ◽

Computer Assisted ◽

Progressive Motility ◽

Frozen Semen ◽

Significant Difference ◽

The Right

Equine semen used for intracytoplasmic sperm injection (ICSI) is typically frozen-thawed and may be of poor quality. To prepare sperm for ICSI, semen is typically centrifuged to remove freezing extender. However, centrifugation can cause damage to sperm, which is especially meaningful if sperm quality is already poor. We evaluated a method for selection of sperm without centrifugation, using a “swim-over” technique, and assessed the effect of pentoxifylline, a phosphodiesterase inhibitor that increases sperm motility in other species. To mimic poor-quality semen, we thawed frozen semen (1×) and re-froze it three additional times (4×). Aliquots (0.25 µL; 50,000 sperm) of 1× or 4× semen were placed at the bottom of the right leg of an “H,” made using 15µL of medium by tracing a template placed below a Petri dish. The medium used (Hanks’ balanced salt solution with 40mg mL BSA and added lactate and pyruvate) contained different concentrations of pentoxifylline (0, 0.5, 1, 2 or 4mgmL−1). One µL of medium was removed from the tip of the left arm of the H after 15 and 30min incubation, and the number of sperm were counted. In a second study, we evaluated the effect of pentoxifylline on sperm motility parameters using computer-assisted sperm motility analysis. After thawing, 1× and 4× semen was washed to remove freezing extender and resuspended in the same medium but with 7mgmL−1 bovine serum albumin (BSA), containing the different pentoxifylline concentrations. In Study 1, the number of collected sperm did not differ significantly for 1× sperm exposed to 0 to 4mgmL−1 pentoxifylline (means of 15 to 23 sperm at 15min, and 18 to 25 sperm at 30min). Similarly, in 4× frozen semen, there was no significant difference in number of collected sperm between 0mgmL−1 and 2 or 4mgmL−1 pentoxifylline concentrations (<1 to 6 at 15 min; 5 to 6 at 30min). In Study 2, at 0min,% total motility was significantly higher in 1 and 2mgmL−1 pentoxifylline than in 0mgmL−1 for 1× sperm (47.8±1.7 and 49.3±1.9, vs. 32.1±3.9, respectively; P=0.018) and significantly higher for 1, 2, and 4mgmL−1 pentoxifylline than for 0mgmL−1 for 4× sperm (3.9±0.9, 5.7±0.4, and 8.2±0.5, vs. 1.2±0.4; P=0.0001). Similar results were found at 15 and 30min for 1×, and at 15min for 4×. Pentoxifylline at 1 to 4mgmL−1 significantly increased the percentage of progressive motility in 1× sperm at 30min (17.8±1.3, 21.8±2.7, and 20.3±1.2, vs. 10.0±0.4; P=0.002) and, at 4mgmL−1, increased the percentage of progressive motility in 4× sperm at 0min (1.43±0.1 vs. 0.2±0.1; P=0.005) and 15min (1.4±0.2 vs. 0.1±0.0; P=0.0001). Exposure of poor-quality semen to pentoxifylline at 4mgmL−1 improved total and progressive motility but did not increase the recovery of motile sperm in a swim-over collection preparation.

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Mouse sperm delivery via cauda epididymis without transport medium

10.7287/peerj.preprints.2542v1 ◽

2016 ◽

Author(s):

Man-Xi Jiang ◽

Mei-Shan Wang ◽

Xiang-Hong Ou ◽

Xue-Jin Chen ◽

Yan Zhu

Keyword(s):

Embryo Development ◽

Sperm Motility ◽

Room Temperature ◽

Sperm Quality ◽

Mouse Sperm ◽

Spermatozoa Motility ◽

Transport Medium ◽

Fertilization Rates ◽

Progressive Motility

This study was aimed to investigate the effects of room temperature (RT, 20-25 oC) and absence of medium during cauda epididymis transport on spermatozoa quality, fertility and embryo development. In the first experiment, fresh sperm from one side of cauda epididymis was used for in vitro fertilization, and another side was delivered at RT or 4-8 oC either with or without M2. In the second experiment, each side of cauda epididymis obtained from the same mouse was individually delivered at RT or 4 oC with or without M2. Finally, sperm motility, progressive motility scores and fertility of fresh spermatozoa or those from transported cauda epididymis, and IVF embryo development were evaluated. Progressive motility scores and fertilization rates were higher in fresh spermatozoa than transported sperm; sperm motility of transported cauda epididymis at 4-8 oC was comparable to fresh spermatozoa, but spermatozoa motility of transported cauda epididymis at RT was inferior to fresh spermatozoa. Spermatozoa motillty of transported cauda epididymis at 4-8 oC with transport medium was much higher than that without transport medium; absence of transport medium did not affect sperm motility of transported cauda epididymis at 4-8 oC but affected sperm motility of transported cauda epididymis at RT. Sperm quality from transported cauda epididymis can be efficiently kept at 4-8 oC, and cauda epididymis transport at 4-8 oC without M2 is more beneficial on keeping their fertility. Moreover cauda epididymis transport at RT without medium could sufficiently produce embryos for obtainning live offsprings.

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Effect of warming temperatures on donkey sperm vitrification in 0.5 mL straws in comparison to conventional freezing

Spanish Journal of Agricultural Research ◽

10.5424/sjar/2019173-14153 ◽

2019 ◽

Vol 17 (3) ◽

pp. e0406 ◽

Cited By ~ 3

Author(s):

Maria Diaz-Jimenez ◽

Jesus Dorado ◽

Cesar Consuegra ◽

Blasa Pereira ◽

Isabel Ortiz ◽

...

Keyword(s):

Plasma Membrane ◽

Sperm Motility ◽

Membrane Integrity ◽

Slow Freezing ◽

Kinematic Parameters ◽

Plasma Membrane Integrity ◽

Progressive Motility ◽

Acrosome Integrity ◽

Sperm Freezing ◽

Conventional Freezing

Aim of study: There is little information about vitrification of sperm in large volumes (up to 0.5 mL). This study aimed to develop the vitrification technique in 0.5 mL straws in donkey sperm, evaluating the effect of three warming temperatures.Area of study: Cordoba, Spain.Material and methods: Ejaculates from five donkeys were divided in four groups: one control subjected to conventional slow freezing (C) and three vitrified in 0.5 mL straws and warmed using different protocols (W1: 37ºC/30s, W2: 43ºC/20s and W3: 70ºC/8s+37ºC/52s). Sperm motility, kinematic parameters, plasma membrane and acrosome integrity were evaluated. Conventional freezing resulted in significantly higher values for total (42.7±19.6%), and progressive motility (30.3±16.7%), plasma membrane (49.1±10.4%) and acrosome integrity (39.6±14.5%) respect to vitrification method.Main results: Values after warming ranged between 0.2-2.8% for total motility; 0.2-2.1% for progressive motility; 5.5-20.0% for plasma membrane integrity and 14.5-29.8% for acrosome integrity in all warming protocols after sperm vitrification. However, no differences were found between W3 and C for kinematic parameters; and W3 resulted in significantly higher values for membrane integrity (20.0±11.0%) in comparison to W1 (5.5±3.6%) and W2 (9.3±8.4%).Research highlights: High warming rates seem to be better for donkey sperm vitrification in large volumes; but this methodology is still not an alternative to conventional sperm freezing.

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Effects of Double Layer Centrifugation on the Improvement of Sperm Quality in Dogs: A Comparative Note among Different Breeds

Annals of Veterinary Science ◽

10.31829/2691-5502/avs2021-4(1)-003 ◽

2021 ◽

pp. 1-12

Keyword(s):

Double Layer ◽

Sperm Motility ◽

Semen Quality ◽

Sperm Quality ◽

Sperm Concentration ◽

Single Layer ◽

Progressive Motility ◽

Prolonged Contact ◽

And Storage ◽

Fresh Semen

The evaluation of sperm quality in the laboratory is essential to improve efﬁciency in assisted reproduction. As in other species, for the dog there are reports that prolonged contact of sperm with some components of seminal plasma is associated with decreased motility and sperm viability. Thus, the centrifugation is a technique widely used to concentrate the spermatozoa and eliminate the supernatant. The purpose of this study was to evaluate the effect of double layer centrifugation on the percentages of total sperm motility and progressive sperm motility of the dog’s semen submitted to the dilution, single layer centrifugation, cooling and storage at 5 °C for 24 and 48 hours. For this purpose, ejaculates of 30 healthy male dogs were evaluated, by taking into account the comparison among the conventional sperm parameters (ejaculate volume, sperm concentration, total sperm motility and sperm progressive motility). The semen samples were examined in standard baseline condition of fresh semen (FS), after dilution (AD), after dilution and single layer centrifugation (SLC), after double layer centrifugation (DLC). According to the different time points, the semen samples were evaluated in baseline conditions, immediately after their collection at (T0), at 24 h (T24) and at 48 h (T48), to evaluate the effect of different treatments on the semen’s quality. Results showed a significant effect of double layer centrifugation on the improvement of total sperm motility and progressive sperm motility percentages of dogs. The use of cooling fresh semen soon after the double layer centrifugation will improve the semen quality up to 48h, with a special emphasis for the percentages of total sperm motility and sperm progressive motility, adding an alternative technical approach to reproductive performance in male breeding dogs.

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Influence of Seasons and Extenders on Quality and Freezability of Gir Bull Semen under Middle Gujarat Climate

THE INDIAN JOURNAL OF VETERINARY SCIENCES AND BIOTECHNOLOGY ◽

10.21887/ijvsbt.v13i03.10632 ◽

2018 ◽

Vol 13 (03) ◽

Cited By ~ 1

Author(s):

D. V. Chaudhari ◽

J. A. Patel ◽

K. K. Hadiya ◽

A. J. Dhami

Keyword(s):

Plasma Membrane ◽

Sperm Motility ◽

Sperm Quality ◽

Membrane Integrity ◽

Winter Season ◽

Egg Yolk ◽

Quality Parameters ◽

Summer Season ◽

Progressive Motility ◽

Bull Semen

The study was conducted to evaluate the seasonal influence (peak winter and summer) and the efficacy of three extenders (egg yolk based TFYG extender and egg yolk free soya bean based commercial extenders Optixcell and Andromed) on quality and freezability of Gir bull semen in Middle Gujarat. Semen ejaculates (6/bull/season, total36) revealed mean ejaculate volume 6.49±0.30 ml, sperm concentration1212.36±58.10 million/ml, progressive motility 74.17±0.78 %, live sperm 81.39±0.80 %, abnormal sperm 7.36±0.31 %, and sperm with intact plasma membrane 81.31±0.98 % and intact acrosome 94.81±0.24 %. Only the progressive sperm motility was significantly (P<0.05) higher(76.39±0.97 % vs. 71.94±1.00 %) with lesser sperm abnormality(6.17±0.37 % vs. 8.56±0.30 %) during winter than in summer. Semen samples split diluted with TFYG, Optixcell and Andromed extenders recorded the overall mean values of progressive sperm motility, livability, abnormality, plasma membrane integrity and acrosomal integrity during winter season as 77.87±0.51, 77.50±0.45, 5.56±0.20,76.02±0.81 and 94.35±0.29 on dilution; 72.41±0.51, 70.50±0.64, 5.96±0.26, 71.20±0.79 and 93.09±0.32 at pre-freeze stage; 41.30 ±0.94, 50.28±1.03, 9.15±0.31, 29.89±0.40 and 90.65±0.40 at post-thaw stage, respectively. The respective values in summer season were 72.13±0.60, 75.50±0.60, 7.48±0.25, 75.61 ±0.55 and 94.09±0.30 on dilution; 65.46±0.66, 69.41±1.05, 8.89±0.28, 69.70±0.66 and 92.63 ±0.33 at pre-freeze stage; 31.48±0.52, 45.09±0.85, 13.48±0.33, 26.85±0.71 and91.26±0.38 at post-thaw stage. The overall mean sperm post-thaw motility/longevity at 0, 30, 60 and 120 min of incubation at 37°C was 41.20±1.51, 35.19±1.47, 28.80±1.75 and 17.50±1.47 % during winter season and 31.57±0.89, 26.20±0.77, 20.37±0.83 and 13.80±0.77% in summer season, respectively. The initial quality as well as freezability of semen in terms of motile, live, normal and HOS reactive sperm including post thaw longevity were better in winter season than in summer season. Further, the values of all the five semen quality parameters studied were comparatively better in Optixcell than TFYG and Andromed extenders with significant differences only in sperm progressive motility in both the seasons.The season x extender interaction was not significant for any of the sperm quality parameters studied.

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Centrifugal force assessment in ram sperm: identifying species-specific impact

Acta Veterinaria Scandinavica ◽

10.1186/s13028-021-00609-8 ◽

2021 ◽

Vol 63 (1) ◽

Author(s):

Marta Neila-Montero ◽

Marta F. Riesco ◽

Mercedes Alvarez ◽

Rafael Montes-Garrido ◽

Juan Carlos Boixo ◽

...

Keyword(s):

Sperm Motility ◽

Assisted Reproductive Technologies ◽

Sperm Quality ◽

Beat Frequency ◽

Movement Pattern ◽

Reproductive Technologies ◽

Centrifugal Forces ◽

Progressive Motility ◽

Species Specific ◽

Ram Sperm

Abstract Background Centrifugation is routinely employed in handling the ejaculates of some species, but it is not part of the commonly used protocols in ram. However, the development and implementation of new assisted reproductive technologies, alternative preservation models based on washing sperm from a cellular ageing-accelerating substance such as the seminal plasma, and basic studies in spermatology is associated with the use of centrifugation. This requires a specific evaluation of the centrifugation protocols considering the species-specific relationship with the potential damage produced by this procedure. No previous studies have determined the effect of different centrifugation forces on ram sperm. Therefore, we aimed to assess the performance of three centrifugal forces (600×g, 3000×g, and 6000×g for 10 min at room temperature) and their effects on ram sperm motility and functionality. Results Sperm motility and functionality parameters were assessed at 0 h and after 2 h of incubation at 37 °C. As expected, a higher cell packaging degree was obtained at high centrifugation forces (P ≤ 0.0001). Cell packaging was unstable at all centrifugal forces. Thus, there was a high cell resuspension rate after less than 2 min. Regarding sperm quality, there was a change in movement pattern of 3000×g and 6000×g centrifuged sperm after 2 h of incubation at 37 °C, characterized by an increase in rapid progressive motility, linearity, straightness, and beat frequency, and a decrease in medium progressive motility, curvilinear velocity, path velocity, and head lateral amplitude. Non-significant differences were obtained among the different treatments concerning the total viability. However, we observed a significant increase (P ≤ 0.05) in the percentage of viable apoptotic sperm in the samples centrifuged at 6000×g at 0 h. Conclusions Centrifugal forces equal to or greater than 3000×g induced some deleterious effects in ram sperm quality, and lower forces did not provide a successful cell packaging degree.

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