86 MOTILITY AND VIABILITY OF BULL SEMEN DURING 24 HOURS OF STORAGE AT 4°C

2006 ◽  
Vol 18 (2) ◽  
pp. 151
Author(s):  
M. E. Carini ◽  
R. Cavia ◽  
G. Larraburu ◽  
G. M. Brogliatti
Keyword(s):  
Storage Time ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Bull Semen ◽  
Multiple Comparison Test ◽  
Straight Line ◽  
Semen Extender ◽  
One Way Anova ◽  
Velocity Average ◽  
Lateral Head

Currently, cryopreservation process of fresh bull semen is carried out between 3 and 6 hours of refrigeration at 4°C post-collection (Hafez, 1989). However, it is sometimes difficult when the cryopreservation process is not available at the site of collection. The objective of this study was to determine seminal motility and viability in samples maintained at 4°C during 24 hours. A total of 98 ejaculates from 23 adult bulls (Angus, Brangus, Braford and Hereford) were collected and diluted in a semi-defined semen extender (Andromed, Minitüb, Tiefenbach, Germany) and stored at 4°C. 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, %), percentage of slow and static cells (SL/ST, %), and viability (VIA, %) were determined by Computer Assisted Semen Analysis (CASA, HTM-ceros 12.1, Berkeley, CA, USA). Measurements were done at 6, 9, 12, and 24 h. The obtained results were analyzed statistically with one-way ANOVA and Dunnet Multiple Comparison Test and are summarized in Table 1. There were no significant differences (P > 0.05) in the VAP, RAPID, or SL/ST during 24 h of storage at 4°C. Measurements were significantly different (P < 0.01) for VSL and VIA at 24 h. Measurements of ALH were increased from 12 h (P < 0.01) and consequently, LIN decreased at the same time (P < 0.01). These results suggest that there are no differences in velocity, except in VSL at the end of the storage time. The type of movement of the spermatozoa change, because ALH increases and the trajectory loses linearity. A decrease in viability suggests that from 24 h of storage, the membrane of the spermatozoa becomes more susceptible. More research needs to be done to evaluate the competence of this time-storage semen in the artificial insemination trial. Table 1. Parameters of motility and viability of semen maintained at 4°C during 24 h This research was supported by Centro Genético Bovino de EOLIA S.A.

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.

scholarly journals 12 CASA PARAMETERS OF FRESH BULL SEMEN COLLECTED BY ARTIFICIAL VAGINA OR ELECTROEJACULATION IN ARGENTINA

2005 ◽  
Vol 17 (2) ◽  
pp. 156 ◽  
Author(s):  
G. Brogliatti ◽  
F. Garcia Migliaro ◽  
R. Cavia ◽  
G. Larraburu ◽  
A. Albrecht
Keyword(s):  
Semen Analysis ◽  
Glass Tube ◽  
Computer Assisted ◽  
Bull Semen ◽  
Straight Line ◽  
Beef Bulls ◽  
Semen Extender ◽  
Artificial Vagina ◽  
Lateral Head ◽  
Motility Parameters

The latest entry in the field of semen evaluation is computer assisted semen analysis (CASA). Its greatest advantages are elimination of the subjective nature of routine semen evaluation and the addition of detailed motion analysis unquantifiable by visual examination. The objective of this study was to evaluate CASA motility parameters of fresh bull semen collected by artificial vagina (AV) or electroejaculation (EE) from a total of 56 beef different bulls. Semen samples from a total of 45 beef bulls were collected by AV from winter to the end of spring (740 collections), and from 11 beef bulls by EE (120 collections) in the same period. First and second AV collections were analyzed as individual data. EE collection was performed only one. Means and standard deviations for each characteristic were calculated, compared, and statistically analyzed. A sample of the collection was diluted 1:20 in a semi-defined semen extender (Andromed, Minitüb, Tiefenbach, Germany) and held in a glass tube at 36°C for 5 min before analysis. The sample was loaded into 20-μm chambers, and six microscope fields from each chamber were analyzed. The following sperm motility parameters were determined with the Ceros 12.1 sperm analyzer (Hamilton Thorne Biosciences, Inc., Beverly, MA, USA) on at least 1000 spermatozoa:concentration (CONC), velocity average path (VAP), velocity straight line (VSL), curvilinear velocity (VCL), amplitude lateral head (ALH), beat cross frequency (BCF), straightness (STR), linearity (LIN), and percentage of rapid or statics cells. There were no significant differences (P > 0.05) in VAP, VSL, VCL, ALH, STR, LIN, and the percentage of rapid and static cells of semen collected by AV or EE. The concentration (sperm/mL) of the AV-collected sperm was significantly higher than for the sperm collected by EE. Results from the analysis indicate that semen collected by artificial vagina have motility characteristics similar to those collected by electroejaculation. More research needs to be done to evaluate motility parameters of frozen/thawed semen collected by electroejaculation and by artificial vagina. This research was supported by Centro Genético Bovino de EOLIA sa Argentina.

scholarly journals Effects of Apigenin and Astragalus Polysaccharide on the Cryopreservation of Bull Semen

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1506
Author(s):  
Hongtao Wang ◽  
Ping Lu ◽  
Chongshan Yuan ◽  
Jing Zhao ◽  
Hongyu Liu ◽  
...  
Keyword(s):  
Semen Analysis ◽  
Computer Assisted ◽  
Astragalus Polysaccharides ◽  
Bovine Semen ◽  
Bull Semen ◽  
Straight Line ◽  
Frozen Semen ◽  
Path Distance ◽  
Straight Line Distance

The purpose of this study was to determine the effects of apigenin and astragalus polysaccharides on the cryopreservation of bovine semen. Apigenin, astragalus polysaccharides, or their combination were added to a frozen diluent of bovine semen. Afterwards, Computer Assisted Semen Analysis (CASA), membrane functionality, acrosome integrity, mitochondrial integrity, CAT, SOD, GSH-Px, MDA, and ROS detection were conducted. The results showed that adding 0.2 mmol/L AP or 0.5 mg/mL APS could improve the quality of frozen sperm. Compared to 0.2 mmol/L AP alone, the combination of 0.2 mmol/L AP and 0.3 mg/mL APS significantly increased the total motility (TM), average path distance (DAP), straight line distance (DSL), average path velocity (VAP), curvilinear velocity (VCL), wobble (WOB), and sperm CAT and SOD levels (p < 0.05), while reducing the ROS and MDA levels (p < 0.05). These results indicated that the addition of 0.2 mmol/L AP or 0.5 mg/mL APS alone has a protective effect on the freezing of bovine semen. Compared to the addition of 0.2 mmol/L AP, a combination of 0.2 mmol/L AP and 0.3 mg/mL APS could further improve the quality of frozen semen.

162 FREEZING OF SEMEN FROM ENDANGERED ASTURIANA DE LA MONTAÑA BULLS IN ZWITTERONIC LIPIDS-BASED EXTENDERS

2008 ◽  
Vol 20 (1) ◽  
pp. 161 ◽  
Author(s):  
C. Tamargo Miguel ◽  
S. S. Pérez-Garnelo ◽  
P. Beltrán Breña ◽  
A. T. Palasz ◽  
J. De la Fuente ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Semen Analysis ◽  
Egg Yolk ◽  
Semen Parameters ◽  
Computer Assisted ◽  
Progressive Motility ◽  
Bull Semen ◽  
Straight Line ◽  
Before And After ◽  
Arcsine Transformation

This experiment was designed to test the efficacy of 2 different preparation protocols of zwitteronic soyabean-origin lipids for the production of lipidsglycerol liposomes for use in bull semen cryopreservation. Lipids liposomes were prepared at 10% concentration in Tris buffer by 1. highpressure homogenization (Panda 2K, Parma, Italy) and then 8% glycerol were added, extender-1 (E-1); Lipids were homogenized together with glycerol, extender-2 (E-2). Bioxcell extender (E-3) was used as control. Semen was collected 3 times from 3 endangered Asturiana de la Monta�a bulls by the means of an artificial vagina. Ejaculates with at least 70% motility were processed further by a standard freezing protocol used in our AI station. Semen was diluted at 37�C with each of the 4 extenders to a concentration of 92 � 106 spermatozoa per mL, cooled to 4�C over 4 h, aspirated into 0.25-mL plastic straws (IMV Technologies, Aigle, France), frozen in a bio-freezer (IMV Technologies) in 3 steps from 4 to –140�C, and then plunged into liquid N2. Straws were thawed in a water bath at 37�C for 30 s. Sperm motility was analyzed microscopically immediately after collection, after dilution, and after 4, 24, 48, and 72 h of storage at 4�C. Post-thaw semen progressive motility was assessed microscopically, and sperm movement characteristics were analyzed by computer-assisted semen analysis (CASA) (SCA�, Microptic, Barcelona, Spain). Data were compared between extenders and bulls by 2-way ANOVA; percentages were transformed by arcsine transformation before ANOVA. Total and progressive sperm motility at 0 h after dilution ranged from 90 to 70% and was not different between extenders and bulls. There was no difference between bulls in total and progressive motility after 24 h of cold storage; however, both were significantly greater (P < 0.05) for Control (62.4 � 14.7 and 41.4 � 14.9) and E-1 (70.1 � 12 and 33.8 � 7.0) extenders than for the E-2 extender (22.5 � 17 and 1.2 � 1.3). Average post-thaw sperm motility was not different between bulls for either extender, but motility for Bioxcell (Control, 48.1 � 14.6%) and E-1 extenders (43.2 � 13.0%) were significantly greater (P < 0.05) than for E-2 extender (18.7 � 8.8%). There were no differences between bulls for all kinematic semen parameters: curvilinear (VCL), straight line (VSL), average path (VAP) velocities, linearity (LIN) and straightness (STR), evaluated by CASA before and after freezing; however, all were lower (P < 0.05) for the E-2 extender and not different between Control and E-1 extenders. Sperm movements derived from heads (VCL) and linearity of sperm(LIN), both closely related to field fertility, were in the range of 90.9 � 2.1 and 63.0 � 5.5 for E-3 (Control) extender; 99.1 � 3.4 and 49.4 � 3.5 for E-1; and 21.8 � 2.2 and 29.9 � 4.0 for E-2. In summary, zwitteronic soyabean lipid liposomes are an effective egg yolk substitute for the cryopreservation of Asturiana de la Monta�a bull semen; however, the homogenization protocol of the lipids-glycerol mixture must be improved.

33 Caffeine improves equine sperm motility after thawing

2019 ◽  
Vol 31 (1) ◽  
pp. 142
Author(s):  
M. A. Lagares ◽  
N. C. Alves ◽  
A. L. A. Guimaraes ◽  
S. B. Luz ◽  
S. A. Diniz ◽  
...  
Keyword(s):  
Sperm Motility ◽  
The Other ◽  
Semen Sample ◽  
Computer Assisted ◽  
Sperm Transport ◽  
Head Displacement ◽  
Straight Line ◽  
Velocity Average ◽  
Lateral Head ◽  
Motility Parameters

The pattern of sperm transport and survival in the mare’s reproductive tract is different between fresh and frozen-thawed semen. A probable reason for this difference is the biophysiological changes in sperm during cryopreservation of equine semen. These changes can impair motility of stallion sperm after thawing. The aim of the present work was to test the effect of different caffeine concentrations on stallion sperm motility after thawing. One ejaculate of 9 stallions was frozen with the INRA82 frozen extender, and after thawing, different caffeine concentrations were added to the semen samples according to the treatments: control INRA82 without caffeine addition (T1), T1+1mM caffeine (T2), T1+2mM caffeine (T3), T1+3mM caffeine (T4), T1+5mM caffeine (T5), T1+7.5mM caffeine (T6), and T1+10mM caffeine (T7). The analysis of sperm motility parameters was performed with a computer-assisted semen analyser in 4 time periods: immediately after semen samples thawing (t0) and 15min (t15), 30min (t30), and 40min (t40) after semen sample thawing. One semen sample of each treatment was thawed, and an aliquot was analysed for the following computer-assisted semen analysis characteristics: velocity curvilinear (VCL; µm s−1), velocity straight line (µm s−1), velocity average path (µm s−1), linearity (%), straightness (%), wobble (%), amplitude of lateral head displacement (µm), beat cross frequency (BCF; Hz), and percentage of total sperm motility (TM) and progressive sperm motility. The statistical analysis was performed with ANOVA and Duncan’s test. The sperm parameters progressive sperm motility, linearity, wobble, and amplitude of lateral head displacement did not differ among the treatments (P&gt;0.05). Immediately after addition (t0) of 5, 7.5, and 10mM caffeine concentrations, an increase of TM was observed (T5: 53.1%; T6: 45.9%; and T7: 47.4%) compared with the other treatments (T1: 37.5%; T2: 36.0%; T3: 36.6%; and T4: 32.3%; P&lt;0.05). Although after 15min of incubation (t15) the TM decreased compared with t0 in T5, T6, and T7 treatments, the percentage was comparable with the other treatments at t15, t30, and t40. The mean value for TM was higher with 5mM caffeine compared with the control group (38.6% v. 34.7%; P&lt;0.05), whereas for the 10mM caffeine treatment velocity straight line (19.9v. 17.1µm s−1), velocity average path (25.6v. 22.9µm s−1), and straightness (75.4v. 72.3%) were higher than the control (P&lt;0.05). For the 5, 7.5, and 10mM caffeine treatments, VCL and BCF were higher than the control (VCL: 33.9, 34.5, 36.8, and 31.5µm s−1, respectively; BCF: 8.1, 8.6, 9.0, and 7.2Hz, respectively). The remaining motility parameters did not differ until 40min after the treatment (P&lt;0.05). In conclusion, the addition of 5, 7.5, and 10mM caffeine concentrations after semen thawing increased TM and most of the sperm motility characteristics. However, given the complexities of sperm transport, capacitation, and so on, further experiments are needed to test whether caffeine treatments could be used to improve the fertilization rate of frozen-thawed equine semen.

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

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.

scholarly journals Correlation between bull fertility and sperm cell velocity parameters generated by computer-assisted semen analysis

2015 ◽  
Vol 63 (3) ◽  
pp. 370-381 ◽  
Author(s):  
Ádám Nagy ◽  
Tassos Polichronopoulos ◽  
András Gáspárdy ◽  
László Solti ◽  
Sándor Cseh
Keyword(s):  
Structural Integrity ◽  
Semen Analysis ◽  
Objective Assessment ◽  
Computer Assisted ◽  
Bull Semen ◽  
Straight Line ◽  
Frozen Semen ◽  
Analysis Computer ◽  
Kinematic Velocity ◽  
Motion Characteristics

Motility is one of the most important characteristics associated with the fertilising ability of spermatozoa indicating their viability and structural integrity. Therefore, the examination of motility constitutes an integral part of semen analysis. Computer-assisted semen analysis (CASA) allows an accurate and objective assessment of different sperm motion characteristics with high repeatability. The aim of this study was to evaluate the different kinematic (velocity) parameters of frozen/thawed bull semen and determine if any of them could be correlated with their fertilising capability after insemination based on the achieved pregnancy rate. Ejaculates from 10 bulls were collected and frozen. The kinematic/velocity parameters of spermatozoa were measured by CASA and compared to the pregnancy results of almost 9,000 females artificially inseminated (AI) with frozen semen of any of the 10 tested bulls. The data of the experiments are summarised mainly with a focus on the effects of individual velocities (curvilinear velocity: VCL, straight-line velocity: VSL, average path velocity: VAP) on fertility rather than on the influence of progressive motility as a whole. We conclude that VAP is the most useful semen motility characteristic which has clinical relevance in the prediction of fertility.

scholarly journals 149 Effects of managing mature beef bulls on divergent planes of nutrition on motility and kinematic properties of fresh and frozen-thawed sperm

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 115-115
Author(s):  
Carl R Dahlen ◽  
Sarah R Underdahl ◽  
Matthew S Crouse ◽  
Kacie L McCarthy ◽  
Cierrah J Kassetas ◽  
...  
Keyword(s):  
Repeated Measures ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Head Displacement ◽  
Time Treatment ◽  
Straight Line ◽  
Frozen Semen ◽  
Beef Bulls ◽  
Kinematic Properties ◽  
Lateral Head

Abstract Fifteen mature beef bulls (BW = 800.4 ± 17.4 kg) were used in a 112-d experiment to evaluate effects of divergent planes of nutrition on motility and kinematic properties of fresh and frozen-thawed semen. Bulls were ranked by BW and randomly assigned to one of two treatments: 1) managed on a positive plane of nutrition (POS, n = 8), or 2) managed on a negative plane of nutrition (NEG, n = 7). Bulls were fed a common diet, adjusted biweekly to achieve targeted weight loss or gain of 12.5% of original BW. On d 112, electroejaculation was used to collect 2 ejaculates from each bull, which were combined. An aliquot of fresh semen was evaluated via computer-assisted semen analysis (CASA; IVOS II, Hamilton Thorne, Beverly, MA, USA) for motility and kinematic properties. Remaining semen was extended and frozen. Frozen semen was thawed for 40 s and held in a heating block at 37°C, then evaluated via CASA at 0 and 3 h post-thaw. Data were analyzed in the MIXED procedure of SAS, with data for post-thaw analysis evaluated as repeated measures in time. Treatment did not influence ejaculate volume or concentration (P ≤ 0.19). In fresh ejaculates no impacts (P ≤ 0.29) of treatment were observed for motility or kinematic properties. In frozen-thawed ejaculates, however, bulls in the NEG treatment had greater (P ≤ 0.02) proportions of motile and progressively motile sperm compared with POS. In sperm classified as motile or progressively motile, NEG had greater (P ≤ 0.002) average path and straight line velocities, and greater (P ≤ 0.05) amplitude of lateral head displacement than POS. Treatment impacts observed in frozen, but not fresh, indicate that sperm metabolism, mitochondrial function, antioxidant capacity, or other factors may be influenced by plane of nutrition resulting in altered motility and kinematic properties.

scholarly journals Ram Semen Cryopreservation Using Egg Yolk or Egg Yolk-free Extenders: Preliminary Results

2019 ◽  
Vol 50 (2) ◽  
pp. 96-103
Author(s):  
M. Ptáček ◽  
M. Stádníková ◽  
F. Savvulidi ◽  
L. Stádník
Keyword(s):  
Semen Analysis ◽  
Egg Yolk ◽  
Field Conditions ◽  
Computer Assisted ◽  
Semen Cryopreservation ◽  
Preliminary Results ◽  
Straight Line ◽  
Custom Made ◽  
Semen Processing ◽  
Semen Extender

Abstract Kinematic parameters of thawed ram semen frozen under field conditions were analysed with the use of two commercial (egg yolk vs egg yolk free) semen extenders in different year-seasonal periods. The semen was collected from Suffolk (n = 2) and Charollais (n = 1) rams kept in private breeding farm on 3 test days (pre-mating, at mating, post mating) during year 2016. Two commercial semen extenders (egg yolk-based BullXcell® or egg yolk-free AndroMed®) were used for semen processing. Processed semen was frozen in 0.25 ml plastic cryostraws using the custom-made portable freezing box for ram semen cryopreservation under field conditions. Thawed semen characteristics were evaluated using computer-assisted semen analysis (CASA) system immediately after thawing and after 2 h of heat incubation (±38°C). Significantly higher total motility (+12.3%), straight line velocity (+5.6 μm s−1), and average-path velocity (+6.9 μm s−1) were detected for the semen processed and cryopreserved using egg yolk-based semen extender. Year-seasonal variation and introducing the ram to service had shown to have a significant effect on the cryopreserved ram spermatozoa. These preliminary results confirmed the feasibility of ram semen processing and cryopreservation under field conditions.

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

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.

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