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

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.

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 Effect of taurine on turkey (Meleagris gallopavo) spermatozoa viability and motility

2018 ◽  
Vol 63 (No. 4) ◽  
pp. 127-135 ◽  
Author(s):  
T. Slanina ◽  
M. Miškeje ◽  
F. Tirpák ◽  
M. Błaszczyk ◽  
R. Stawarz ◽  
...  
Keyword(s):  
Meleagris Gallopavo ◽  
Lower Percentage ◽  
Computer Assisted ◽  
Viability Assessment ◽  
Spermatozoa Motility ◽  
In Vitro Incubation ◽  
Time Periods ◽  
Casa System ◽  
Motility Parameters

The effect of taurine on the turkey spermatozoa motility and viability during the in vitro incubation was assessed. Experimental samples were prepared by diluting the raw semen in nine different concentrations of taurine – from 10 mg/ml to 0.078125 mg/ml. The motility parameters were evaluated by the CASA system (Computer Assisted Semen Analyser) using the program Sperm Vision<sup>®</sup> and for spermatozoa viability assessment the eosin-nigrosin staining was performed. Selected parameters were evaluated at six time periods: 0, 1, 2, 3, 4, and 5 h at 5°C and 41°C. At 5°C, a significantly lower percentage of motility and progressive motility was detected only in the samples with the highest concentration of taurine (10 mg/ml) at time 0 and 1. After 2 h of incubation a significant preventive effect of taurine on spermatozoa parameters was observed. The tendency of the taurine effect on motility parameters was different during the in vitro incubation at 41°C. Significantly lower values of motility parameters were detected in all experimental samples in comparison to the control after 5 h. The analysed concentrations of taurine did not significantly affect viability of turkey spermatozoa during all time periods. A higher percentage of dead spermatozoa were observed at 41°C (4.87–9.90%) if compared to 5°C (2.12–4.88%). The results indicated that the addition of taurine (from 2.5 to 7.5 mg/ml) to turkey spermatozoa positively affected the monitored spermatozoa parameters incubated at 5°C.

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.

67 EFFECTS OF ANTIOXIDANTS LACTOFERRIN AND CATALASE ON STALLION FROZEN SEMEN

2015 ◽  
Vol 27 (1) ◽  
pp. 126
Author(s):  
H. S. Martins ◽  
M. F. Brito ◽  
I. B. M. Sampaio ◽  
R. Stahlberg ◽  
M. R. Souza ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Skim Milk ◽  
Straight Line ◽  
Frozen Semen ◽  
Significant Difference ◽  
Sperm Membrane ◽  
Positive Effect ◽  
Artificial Vagina ◽  
Motility Parameters

During cryopreservation, the sperm were submitted to an increased generation of reactive oxygen species. Furthermore, because of the large portion of seminal plasma removal, there is a decrease of sperm antioxidant protection. Addition of antioxidants proteins found in seminal plasma, such as lactoferrin (Lf) and catalase (Cat), to the freezing semen extenders could protect the sperm during cryopreservation. Lactoferrin is a transferrin, which prevents the hydroxyl radicals generation, and Cat plays an antioxidant role. The aim of this study was to determine the effects of Lf and Cat supplementation to the INRA 82 freezing extender (Battelier et al. 1997) on sperm motility parameters and membrane functionality of stallion frozen semen. Semen from 6 stallions was collected with an artificial vagina, diluted with Kenney extender (1 : 1), and centrifuged (500 × g, 10 min). The supernatant was discarded, and sperm number per milliliter was calculated. Semen was resuspended with 3 extenders to 100 × 106 sperm mL–1. The treatments were distributed in (F1) control, INRA 82 freezing extender (Battelier et al. 1997), (F2) F1+ 500 μg mL–1 lactoferrin, and F3) F1 + 200 IU mL–1 catalase. Semen samples were packaged in 0.5-mL straws and cooled to 5°C (0.27°C min–1). For semen freezing, the straws were laid over the LN vapor for 20 min and plunged into the LN. The straws were thawed at 37°C for 30 s. Motility parameters of frozen semen were determined using a computer sperm cell analysis, and sperm membrane functionality was assessed by the hyposmotic swelling test (Lagares et al. 1998). The data were analysed using Friedman test using stallion as a block. A probability of P < 0.05 was considered significant. There was no significant difference between the percentage of total sperm motility (median, minimum-maximum value; F1: 29.9, 11.0–82.7; F2: 49.8, 7.7–55.2; F3: 39.8, 5.7–92) and progressive sperm motility (F1: 7.1, 3.2–23.3; F2: 13.4, 2.6-22.4; F3: 15.6, 1.1–29.6), and functional sperm membrane (F1: 26.7, 14.7–56.2; F2: 50.5, 15.7–61.7; F3: 46.6, 13.8–50.9) with regard to the treatment. However, the velocity parameters: velocity average path (F1: 29.3, 22.1–33.80; F2: 34.6, 24.8–44; F3: 35.7, 18.2–42.6), velocity curvilinear (F1: 36.9, 30.5–45.1; F2: 42.5, 34.7–51; F3: 44.6, 25.5–50.9), and velocity straight line (F1: 23.4, 17–3.60; F2: 28.9, 18.8–38.2; F3: 26.6, 13.6–37.2) in the treatment with Lf (F2) were higher compared with the control (F1; P < 0.05). These results corroborate with studies reporting the lack of positive effect on equine sperm motility when antioxidants were added to skim milk-based extenders. Although the addition of Lf or Cat to skim milk-based extenders did not improve the motility sperm characteristics and sperm membrane functionality, more studies about the positive effect of Lf on the velocity parameters are necessary. Lactoferrin could then play an important role on the oxidative metabolism, which provides energy to the sperm movement.Acknowledgments to the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brazil, for the financial support.

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 Effect of the addition of sodium caseinate on the viability of cryopreserved buffalo semen

2020 ◽  
pp. 2209-2218
Author(s):  
Fernando Evaristo da Silva ◽  
Jaqueline Candido Carvalho ◽  
Camila de Paula Freitas Dell'Aqua ◽  
Frederico Ozanam Papa ◽  
Marc Roger Jean Marie Henry ◽  
...  
Keyword(s):  
Cell Damage ◽  
Membrane Integrity ◽  
Sodium Caseinate ◽  
Egg Yolk ◽  
Freezing Process ◽  
Computer Assisted ◽  
Semen Cryopreservation ◽  
Sperm Analysis ◽  
Frozen Semen ◽  
Buffalo Semen

The use of cooled semen in artificial insemination operations results in higher pregnancy rates than the use of frozen semen. This result seems to be related to the more severe damage triggered by the freezing process than that observed during refrigeration. Due to its ability to bind to sperm-binding proteins and calcium ions, sodium caseinate has been studied as a substance capable of preventing early sperm capacitation, a significant cause of the decreased pregnancy rate resulting from the use of frozen semen. The first objective of this study was to evaluate whether a commercial egg yolk diluent developed for frozen bovine semen could be used for buffalo semen cryopreservation; the second objective was to investigate the effect of this diluent in combination with sodium caseinate during the procedures of buffalo sperm cryopreservation using flow cytometry and computer-assisted sperm analysis. In the first part of the study, comparing the results of spermatic kinetics and plasma and acrosomal membrane integrity, it was observed that the freezing process resulted in more cell damage than the cooling process. In the second part of the study, no effects of the addition of sodium caseinate to the egg yolk diluent were observed. From the results of the present study, it was possible to conclude that the egg yolk-based diluent was suitable for buffalo semen cryopreservation and that the addition of sodium caseinate did not decrease the harmful effects related to seminal cryopreservation.

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.

Rapid freezing of the mouse blastocyst: effects of cryoprotectants and of time and temperature of exposure to cryoprotectant before direct plunging into liquid nitrogen

1991 ◽  
Vol 3 (2) ◽  
pp. 175 ◽  
Author(s):  
R Li ◽  
A Trounson
Keyword(s):  
Liquid Nitrogen ◽  
Room Temperature ◽  
Rapid Freezing ◽  
High Concentration ◽  
Freezing And Thawing ◽  
Initial Exposure ◽  
Survival And Development ◽  
High Concentrations

This study investigates the effects of time and temperature of exposure to a high concentration (4.5 M) of dimethyl sulfoxide (DMSO), glycerol, 1,2-propanediol (PROH), or a mixture of DMSO and glycerol (DG) in a solution containing 0.25 M sucrose, on the survival and development of rapidly frozen mouse blastocysts. Embryos had significantly (P less than 0.01) higher rates of survival and development when exposed to cryoprotectant at 0 degree C compared with room temperature. The time of exposure to cryoprotectant at either 0 degree C or room temperature before being plunged into liquid nitrogen significantly (P less than 0.01) affected the survival and development of frozen-thawed embryos. Survival and development of blastocysts in vitro and in vivo was significantly (P less than 0.05) higher when exposed at 0 degree C for 10 min to DG, DMSO and glycerol than to PROH. It is concluded that, unlike early-cleavage stage embryos, blastocysts need to be equilibrated at a low temperature (0 degree C) with high concentrations of cryoprotectant before rapid freezing. Exposure of blastocysts to 4.5 M cryoprotectant and 0.25 M sucrose at room temperature either was toxic or else markedly reduced their viability after freezing and thawing, depending on the duration of the initial exposure.

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