76 COMPUTER-ASSISTED SPERM ANALYSIS OF FROZEN SPERM MOTILITY AFTER DIFFERENT TIMES OF EXPOSURE AT 15°C

2008 ◽  
Vol 20 (1) ◽  
pp. 119
Author(s):  
A. Garcia Guerra ◽  
M. P. Etcheverry ◽  
D. Rodriguez ◽  
G. Larraburu ◽  
G. M. Brogliatti
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Room Temperature ◽  
Cell Damage ◽  
Semen Analysis ◽  
Beat Frequency ◽  
Computer Assisted ◽  
Frozen Semen ◽  
Significant Difference ◽  
Casa System

One of the key factors for successful long-term cryopreservation in liquid nitrogen is maintaining the samples at –130°C or lower at all times to avoid cell damage (Barth 1991 Proc. 10th Ann. Conv. Am. Embr. Transf. Assoc., 20–26). Previous data indicated that exposure of the semen straw to ambient temperature for more than 15 s can raise the temperature above –130°C and reduce sperm motility, as determined by subjective evaluation (Berndtson et al. 1976 Proc. 6th NAAB Tech. Conf. Artif. Insem. Reprod., 51–60). The computer-assisted semen analysis (CASA) system provides an opportunity to assess multiple motility characteristics on a semen sample objectively and with high repeatability. An experiment was designed to evaluate the effect of exposing frozen semen in 0.5-mL straws to room temperature for 15, 30, 60, or 120 s on motility characteristics assessed by CASA system. Twenty-eight ejaculates from different bulls (19 Angus, 7 Hereford, 1 Brangus, 1 Shorthorn) were diluted using a chemically semi-defined media (Andromed, Minitüb, Tiefenbach, Germany) and frozen in an automatic freezer (Digicool, IMV, Paillette Crista, France). Five frozen straws per bull were used, one for each time of exposure and one as control (0 s = 0 time). Straws were exposed to room temperature (15°C ± 0.78) for different times and then placed back into liquid nitrogen. Semen thawing was conducted in a water bath at 37°C for 1 min. Motility characteristics were evaluated by the IVOS SpermAnalyzer (Hamilton Thorne Research, Beverly, MA, USA). Two chambers of 20-μm depth and 5 fields per chamber were analyzed (30 frames/0.5 s for each field). Seven motility parameters were evaluated: motile sperm (%), progressive sperm (%), VAP (path velocity, μm s–1), VCL (track speed, μm s–1), ALH (lateral amplitude, μm), BCF (beat frequency, Hz), and LIN (Linearity, %). The Kruskal–Wallis test was used to compare variables among groups, and results are shown in Table 1. There is a significant difference (P < 0.05) in the % of motile and progressive sperm when time of exposure was increased. There was a drastic and significant reduction in the percentage of motile and progressive sperm when exposure to 15°C was longer than 30 s. The live cells had similar motile characteristics: VAP, VCL, ALH, BCF, and LIN. In conclusion, sperm motility would be affected if straws are exposed for more than 30 s. More research should be done to test higher room temperatures, detect viability effects, and determine pregnancy rates after AI. Table 1. CASA of frozen sperm motility characteristics at different times of exposure at 15°C This research was supported by Centro Genetico Bovino Eolia S.A.

84 COMPUTER-ASSISTED SPERM ANALYSIS OF FROZEN SPERM MOTILITY AFTER REPEATED EXPOSURES TO ROOM TEMPERATURE

2010 ◽  
Vol 22 (1) ◽  
pp. 200
Author(s):  
A. Alvaro Garcia Guerra ◽  
G. M. Brogliatti
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Room Temperature ◽  
Cell Damage ◽  
Beat Frequency ◽  
Computer Assisted ◽  
Initial Exposure ◽  
Frozen Semen ◽  
Casa System ◽  
Motility Parameters

The key factorin long-term cryopreservation is the very low temperature of liquid nitrogen. Several studies suggest temperatures should be maintained at -130°C or less to avoid cell damage. Damage due to initial exposure may not be overt; however, after repeated exposures a reduction in postthaw viability may become evident (Barth A 1991 Proc. 10th Annu. Conv. Am. ET Assoc, 20-26). The CASA system provides an opportunity to assess multiple motility characteristics on a semen sample objectively and with high repeatability. An experiment was designed to evaluate the effect that repeated exposure of frozen semen in 0.5-mL straws during 15 s to room temperature produces on motility characteristics assessed by CASA system. Groups were formed according to the number of exposures per straw; groups were as follows: 0, 3, 5, and 10 times of exposure during 15 s. Thirty-two ejaculates from different bulls (15 Angus, 3 Hereford, 8 Brangus, 3 others) were diluted using a chemically semi-defined media (Andromed, Minitub, Germany) and frozen in an automatic freezer (Digicool, IMV, Paillette Crista, France). Four frozen straws per bull were used, one for each group. Straws were exposed to a room temperature (15°C ± 1.28) and then placed back into liquid nitrogen. Semen thawing was conduced in a water bath at 37°C during 1 min. Motility characteristics were evaluated by the IVOS Sperm Analyzer (Hamilton Thorne Research). Two chambers of 20 μm depth and 5 fields per chamber were analyzed (30 frames/0.5 s for each field). Seven motility parameters were evaluated: % of motile sperm; % of progressive sperm; VAP (path velocity, μms-1); VCL (track speed, μm/s); ALH (lateral amplitude, μm); BCF (beat frequency, Hz); and LIN (linearity, %). The Kruskal Wallis test was used to compare variables among groups, and results are shown in Table 1. The average temperature inside the straw after 15 s of exposure was of -122.6°C. No difference (P > 0.05) was found among the groups for any of the 7 motility parameters. In conclusion, sperm motility seems not to be affected if straws are exposed up to 10 times during 15 s to room temperature. More research should be done to test higher room temperatures and pregnancy rates after AI. Table 1.CASA parameters of frozen sperm after different numbers of exposures at 15°C

91 MOTILITY (CASA) AND VIABILITY OF FROZEN SPERM AFTER DIFFERENT TIMES OF EXPOSURE AT 15°C

2010 ◽  
Vol 22 (1) ◽  
pp. 204
Author(s):  
A. Garcia Guerra ◽  
M. G. Lüssenhoff ◽  
G. M. Brogliatti
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Room Temperature ◽  
Bos Taurus ◽  
Cell Damage ◽  
Subjective Evaluation ◽  
Beat Frequency ◽  
Bos Indicus ◽  
Frozen Semen ◽  
Live Sperm

One of the key factors for successful long-term cryopreservation in liquid nitrogen is maintaining the samples at -130°C or lower at all times to avoid cell damage (Barth A 1991 Proc. 10th Annu. Conv. Am. ET Assoc., 20-26). Previous data reported that exposure of semen straw to ambient temperature for more than 15 s can raise the temperature above -130°C and could reduce sperm motility by subjective evaluation (Berndston et al. 1976 Proc. 6th NAAB Tech. Conf. Artif. Insem. Reprod., 51-60). The CASA system provides an opportunity to assess multiple motility characteristics on a semen sample objectively and with high repeatability. Two experiments were designed to evaluate the effect of exposing frozen semen in 0.5-mL straws for 15, 30, or 60 s to room temperature on motility characteristics assessed by CASA and viability parameters by vital stain, HOS test, and acrosome integrity. Thirty-three ejaculates from different bulls (88% British breeds) were used for CASA evaluation, and 12 ejaculates were from other bulls (7 Bos indicus and 4 Bos taurus) were used for viability evaluation. All ejaculates were diluted using a chemically semi-defined media (Andromed, Minitub, Germany) and frozen in an automatic freezer (Digicool, IMV, France). Five frozen straws per bull were used, one for each time of exposure and one as control (0 s = 0 time). Straws were exposed to room temperature (15°C ± 1.78) for different times and then placed back into liquid nitrogen. Semen thawing was conduced in a water bath at 37°C during 1 min. Motility characteristics were evaluated by the IVOS Sperm Analyzer (Hamilton Thorne Research). Two chambers of 20 μm depth and 5 fields per chamber were analyzed (30 frames/0.5 s for each field). Six motility parameters were evaluated: % of motile sperm; % of progressive sperm; VAP (path velocity, μm s-1); ALH (lateral amplitude, μm); BCF (beat frequency, Hz); and LIN (linearity, %). Viability characteristics were evaluated by % of live sperm (eosin-nigrosin); % positive to HOS test, and % of intact acrosome (Giemsa stain). A nonparametric AOV (Kruskal Wallis) test was used to compare variables among groups, and results are shown in Table 1. There was a reduction (P < 0.05) in the percentage of motile and progressive sperm when exposure to 15°C was longer than 30 s. The alive cells have similar motile characteristics as VAP, VCL, ALH, BCF, and LIN. The viability of spermatozoa was reduced (P < 0.05) when they were exposed to room temperature beyond 30 s. Also, a lower proportion of positive spermatozoa for HOS test was detected for exposures beyond 15 s. In conclusion, these results suggest sperm motility and viability would not be affected if straws are exposed up to 30 s to 15°C. Further study should be done regarding viability tests. Table 1.Motility and viability parameters of exposed frozen semen

scholarly journals 12PREDICTION OF BULL FERTILITY BY COMPUTER ASSISTED SEMEN ANALYSIS

2004 ◽  
Vol 16 (2) ◽  
pp. 128 ◽  
Author(s):  
S. Cseh ◽  
T. Polichronopoulos ◽  
L. Solti
Keyword(s):  
Sperm Motility ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Reproductive Process ◽  
Return Rates ◽  
Frozen Semen ◽  
Chi Square Analysis ◽  
Fertilizing Capability ◽  
Weighted Values

Sperm motility is clearly essential for fertilization both in vivo and in vitro. Motility is necessary for successful sperm transport, a step that is bypassed with in vitro fertilization. Recently, increasing attention has been paid to the objective evaluation and characterization of sperm motility more than simply determining the total proportion of motile spermatozoa. The purpose of computerassisted semen analysis (CASA) is to provide values for sperm concentration and sperm motility more rapidly and accurately than those obtained with traditional semen analyses methods. The objective of our experiment was to investigate the effect of specific aspects of sperm movement, such as the velocity of progression and the actual pattern of movement, to the fertilizing capability of sperm. Frozen semen samples of 10 HF breeding bulls were used in the study. For the motility analyses, Medealab CASA system (Medealab, Germany, Ver. 4.1) was used, and the velocity parameter of VCL (curvalinear velocity, μms−1), VSL (straight line velocity, μms−1), and VAP (average path velocity, μms−1) were evaluated and compared with the Day 30 and 75 non−return rates (NR30 and NR75). For every sample, a total of 10 fields were examined for 8s using a disposable 20 micron capillary chamber (CellVision, USA) giving a total of 1165 to 2831 cells evaluated. Chi square analysis, analyses of variance and linear correlation coefficient was applied to the statistical evaluation and comparison of the results. Data are based on weighted values. From the same batch of the analyzed frozen semen, a total of 8099 females were inseminated in more than 100 farms with a total of 6590 animals being positive for pregnancy at Day 30 and 4525 animals at Day 75. Within the bulls, differences were found in the values of NR30 and NR75 (P&lt;0.05). Our data indicate very strong differences between the males’ NR30 and NR75 values (NR30: 65.6%±13.04 to 79.6%±11.17; P&lt;0.001 and NR75: 37.8%±10.38 to 58.3%±15.53; P&lt;0.001) reflecting the individual differences in the fertilizing capability of the males. All velocity parameters show very high correlation with strong significance both non−return rates but the best values belong to VAP (NR30 and NR75; P&lt;0.02). Our data indicate that the bulls with lower VCL (25.51±33.04 to 79.54±58.03), VSL (11.35±19.45 to 36.36±35.71), and VAP (12.67±19.06 to 41.75±34.45) values showed lower fertilization rates both at NR30 and NR75. Computer and video technologies have advanced rapidly in recent years; thus the capability and accuracy of the latest versions of CASA systems are considerably better and they give more information about the different motion characteristics of spermatozoa. Because of the vital role of sperm motility in the reproductive process, such systems will enable us to move into a new era of diagnostic andrology and predict the fertilizing capability of semen. Supported by NKFP-Grants 4/040/2001 and 4/031/2001.

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

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 (&lt;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.

scholarly journals Assessment of an iPad-based Sperm Motility Analyzer for Determination of Canine Sperm Motility

2021 ◽  
Author(s):  
Evelyn Bulkeley ◽  
Christine Collins ◽  
Azarene Foutouhi ◽  
Kris Gonzales ◽  
Heather Power ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Progressive Motility ◽  
Alternative Measurement ◽  
Positive Correlations ◽  
Casa System ◽  
Progressive Assessment

Abstract The objective of this study was to evaluate the repeatability and accuracy of canine sperm motility (total and progressive) assessment with a tablet-based Canine iSperm ® instrument compared to computer-assisted sperm analysis (CASA). The experiment used fresh and frozen/thawed canine semen samples for comparisons of semen analysis parameters (concentration, total motility, and progressive motility) between a CASA system, iSperm ®, and NucleoCounter ® SP-100 ™ (concentration) instruments. Spearman’s Rho correlational analysis was used to identify significant associations between motility assessment methods. Significant positive correlations were found between CASA assessment and iSperm ® for both progressive and total motility measurements. We also determined the coefficient of variation (CV) for repeatability of sample analysis for iSperm ® and CASA for fresh sperm, wherein each sample was assessed 10 times on both devices. For fresh and frozen-thawed samples, concentration assessment by iSperm ® showed high variability (CV= 19.9 ± 1.5%). For iSperm ® assessment of total and progressive motility, the CV’s were 6.3 ± 0.5% and 10.7 ± 0.8%, respectively. The results indicate that the iSperm ® application offers an accurate and alternative measurement of motility to traditional CASA analysis, though caution should be taken when assessing concentration due to the high CV observed in this study.

Effect of separate and combined exposure of selenium and diazinon on rat sperm motility by computer assisted semen analysis

2016 ◽  
Vol 38 ◽  
pp. 144-149 ◽  
Author(s):  
Robert Toman ◽  
Svatoslav Hluchy ◽  
Michal Cabaj ◽  
Peter Massanyi ◽  
Shubhadeep Roychoudhury ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Combined Exposure

Impact of supplementing commercial rabbit pellets with fresh Moringa oleifera leaves on semen characteristics in rabbit bucks under tropical climate in Trinidad

2021 ◽  
Author(s):  
Krishna Mohan Kumar
Keyword(s):  
Sperm Motility ◽  
Moringa Oleifera ◽  
Sperm Count ◽  
Control Group ◽  
Computer Assisted ◽  
Group Iii ◽  
Group I ◽  
Significant Difference ◽  
Moringa Oleifera Leaves ◽  
The Impact

Objective This study aimed to evaluate the impact of the dietary supplement of Moringa oleifera leaves (MOL) on semen quality and characteristics in rabbits. Methods Eighteen (n=18) breeding bucks of New Zealand white, of similar age group, were used for the study. Three feeding regimes, (i) 100% commercial rabbit pellets (CRP)-Group I (ii) 90% CRP + 10% fresh MOL on a dry matter (DM) basis – Group II and (iii) 80% CRP + 20% fresh MOL on a DM basis – Group III, were adopted and the trial continued for 21 days. After adaptation to the diet, semen was collected from each buck and subjected to evaluation using a computer-assisted semen analyser. Results In Group III, the sperm count, normal sperm morphology, and sperm motility increased (52.0%) in comparison with the control (Group I; 50.1%). The inclusion of 20% Moringa oliefera in the diet (Group III) caused a significant increase (P<0.05) in semen concentration (Control =136.2 M/mL; Group III=297.2 M/mL). There was no significant difference (P>0.05) in sperm motility and semen volume among the groups. Conclusion The results suggest that supplementing commercial rabbit pellets with 20% fresh Moringa oliefera leaves on a DM basis can improve the quality and characteristics of semen in breeding bucks.

84 EVALUATION OF BULL SEMEN AT 1 H VS. 48 H OF POST-FROZEN STABILIZATION TIME

2006 ◽  
Vol 18 (2) ◽  
pp. 150
Author(s):  
G. M. Brogliatti ◽  
G. Larraburu ◽  
R. Cavia ◽  
M. E. Carini
Keyword(s):  
Liquid Nitrogen ◽  
Artificial Insemination ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Stabilization Time ◽  
Bull Semen ◽  
Straight Line ◽  
Semen Extender ◽  
Lateral Head

The process of cryopreservation of bull semen in liquid nitrogen at −196°C is usually carried out after 3 to 6 h of refrigeration at 4°C post-collection. To guarantee the quality of the final product, the frozen straws are evaluated after cryopreservation. The seminal samples are usually stabilized during 48 h before being analyzed (Hafez, Reproduction and Artificial Insemination in Animals, 1989); this would retard the possible commercialization. The objective of the present study was to determine motility parameters and viability of semen doses stabilized by 1 h or more than 48 h in liquid nitrogen at −196°C. A total of 122 ejaculated from 23 different adult bulls (Angus, Brangus, Braford, and Hereford) were evaluated in an artificial insemination center between January and April 2005. The semen was diluted in a semi-defined semen extender (Andromed, Minitub, Germany) and frozen in an automatic freezer (Digicool, IMV, France). Parameters of velocity average path (VAP, μm/s), velocity straight line (VSL, µm/s), amplitude lateral head (ALH, µm), linearity (LIN, %), percentage of rapid cells (RAPID, %), and viability (VIA, %) were determined by Computer Assisted Semen Analysis (CASA, HTM-ceros 12.1, Berkeley, CA, USA). The obtained results were analyzed statistically with T Student and are summarized in Table 1. The results indicate that there is no difference in the velocity of the spermatozoa evaluated 1 h or 48 h post-frozen. There is no difference in VAP, VSL, movement of amplitude lateral head (ALH), or linearity (LIN). The percentage of viable spermatozoa was not affected in either group. Statistical analysis indicates that there is no difference (P > 0.05) in any of the evaluated parameters. The results demonstrate that spermatic motility and viability of frozen bull semen could be evaluated before 48 h post-frozen. This allows reduction of the time between freezing and evaluation and immediate availability of the bull straws. Table 1. Parameters of motility and viability at 1 h vs. 48 h of post-frozen stabilization time This research was supported by Centro Genético Bovino EOLIA S.A.

77 ADDITION OF CHOLESTEROL IN FRESH GOAT SPERM IMPROVES CRYOSURVIVAL RATES

2013 ◽  
Vol 25 (1) ◽  
pp. 186
Author(s):  
B. G. Silva ◽  
E. A. Moraes ◽  
C. S. Oliveira ◽  
W. D. Ferrari Junior ◽  
W. C. G. Matos ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Phase Contrast ◽  
Room Temperature ◽  
Petri Dish ◽  
Egg Yolk ◽  
Irreversible Damage ◽  
Hypoosmotic Swelling Test ◽  
Contrast Microscopy ◽  
Working Solution

Cryopreservation causes irreversible damage to goat sperm membranes, measured by a loss of motile and functional normal cells, compared with fresh sperm. The objective of this study was to determine if the addition of cholesterol-loaded cyclodextrin (CLC) to goat semen improved sperm cryosurvival. The CLC was prepared as described by Purdy and Graham (2004 Cryobiology 48, 36–45) with some modifications: 200 mg of cholesterol were dissolved in 1 mL of chloroform and 1 g of methyl-beta-cyclodextrin was dissolved in 2 mL of methanol. A 0.45-mL aliquot of the cholesterol solution was added to the cyclodextrin solution, after which the mixture was poured into a glass Petri dish and the solvents allowed to evaporate on a warm plate for 24 h. The resulting crystals were removed from the dish and stored at 22°C. A working solution of the CLC was prepared by adding 50 mg of CLC to 1 mL TALP at 37°C. Thirty ejaculates from 5 bucks were collected, diluted 1 : 1 in Tris diluent, divided into 7 equal aliquots, and centrifuged at 800g for 10 min. The sperm pellets were resuspended in Tris diluent, to which 0, 0.75, 1.5, 3.0, 4.5, 6.0, or 7.5 mg of CLC/120 million sperm were added. All treatments were incubated for 15 min at room temperature and then cooled to 4°C over 2 h. The samples were then diluted with Tris-egg-yolk diluent containing 2% glycerol, and the sperm were packaged into 0.5-mL straws, frozen in static liquid-nitrogen vapour for 20 min, and plunged into liquid nitrogen. Straws were thawed in 37°C water for 30 s, extended in Tris, and analyzed using optic microscopy. To test thermal resistance, after thawing, 0.5 mL of semen from each treatment were placed in 1.5-mL Eppendorf tubes in a water bath at 37°C for 3 h. At 0, 60, 120, and 180 min, subsamples were evaluated for sperm progressive motility. A hyposmotic test was also conducted by adding 10 µL of sperm to 2 mL of each solution and incubating them for 1 h/37°C. Sequentially, 20 µL of sperm was diluted in hypoosmotic solution (150 mOsm), and the samples were evaluated using phase-contrast microscopy. A total of 100 spermatozoa were counted in at least 5 different fields, and sperm tails were classified as either noncoiled or coiled. Data were analyzed using ANOVA, and treatment means were separated, using the SNK test at 5% probability. The sperm motility (50.4, 33.8, and 22.5%) was significantly higher for sperm treated with 0.75 mg of cholesterol after 0, 60, and 120 min of incubation after thawing, when compared with other treatments. No treatment differences in the hypoosmotic swelling test were observed. The addition of 0.75 mg of cholesterol to fresh goat semen improved sperm motility after cryopreservation for up to 3 h. Supported by FACEPE and CAPES.

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