Understanding motility dynamics of crossbred bull spermatozoa when analyzed by Computer Assisted Semen Analyzer (CASA)

2017 ◽  
Author(s):  
Saroj Rai ◽  
S. Tyagi ◽  
M. Kumar ◽  
M. Karunakaran ◽  
M. Mondal ◽  
...  
Keyword(s):  
Circular Path ◽  
Computer Assisted ◽  
Low Velocity ◽  
Head Displacement ◽  
Semen Collection ◽  
Straight Line ◽  
Slow Moving ◽  
The One ◽  
Kinetics Of ◽  
Lateral Head

The study was conducted to understand sperm kinetics of Frieswal bull spermatozoa using Computer Assisted Semen Analyzer (CASA). Fifty bull ejaculates were collected from ten healthy bulls that were in routine semen collection. The semen samples were diluted in Tris buffer at a concentration of 25x106 spermatozoa/ ml for analysis. Rapidly moving spermatozoa represented the one with better velocity and progressiveness while spermatozoa with medium motility had low velocity with short distance travelled in spite of its ability to move in a straight line (straightness, STR >70 %). The slow moving cells had good head and flagella movement but they followed a circular path with straight line velocity (VSL mm/ sec), linearity (LIN %) and straightenss (STR %) of 10.91, 9.00 and 22.54, respectively. Results indicated that individual sperm cells tracked by CASA as rapid, medium and slow motile were highly variable (p>0.001). However, the sperm motility between bulls varied (p>0.05) only in lateral head displacement (ALH, mm) and beat cross frequency (BCF, Hz).

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

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
P. Perumal ◽  
S. K. Srivastava ◽  
S. K. Ghosh ◽  
K. K. Baruah
Keyword(s):  
Objective Analysis ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Progressive Motility ◽  
Head Displacement ◽  
Straight Line ◽  
Computer Assisted Sperm Analysis ◽  
Lateral Head ◽  
Motility Parameters

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

scholarly journals Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields

Animals ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 1993
Author(s):  
Sabrina Gacem ◽  
Jaime Catalán ◽  
Anthony Valverde ◽  
Carles Soler ◽  
Jordi Miró
Keyword(s):  
High Resolution ◽  
Sperm Motility ◽  
Frame Rate ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Head Displacement ◽  
Straight Line ◽  
Counting Chamber ◽  
Computer Assisted Sperm Analysis ◽  
Lateral Head

In order to optimize the donkey sperm motility analysis by the CASA (Computer Assisted Sperm Analysis)-Mot system, twelve ejaculates were collected from six jackasses. Capillary loaded chamber (CLC), ISAS®D4C depths 10 and 20 µm, ISAS®D4C Leja 20 and drop displacement chamber (DDC), Spermtrack® (Spk) depths 10 and 20 µm were used. Sperm kinematic variables were evaluated using each chamber and a high-resolution camera capable of capturing a maximum of 500 frames/second (fps). The optimum frame rate (OFR) (defined according to curvilinear velocity—VCL) was dependent on chamber type. The highest OFR obtained was 278.46 fps by Spk20. Values for VCL, straight-line velocity (VSL), straightness (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) were high in DDC and 10 µm depth. In both DDC 10 and 20 µm, the sperm velocities (VCL, VSL, VAP) and ALH values decreased significantly from the centre to the edges, while Wobble and BCF increased. No defined behavior was observed along the CLC. However, all the kinematic variables had a higher value in a highly concentrated sample, in both chamber types. In conclusion, analyzing a minimum of nine fields at 250 fps from the centre to the edges in Spk10 chamber using a dilution of 30 × 106 sperm/mL offers the best choice for donkey computerised sperm motility analysis.

14 FREEZING OF DONKEY SEMEN AFTER 24 HOURS OF COOL STORAGE: PRELIMINARY RESULTS

2013 ◽  
Vol 25 (1) ◽  
pp. 154 ◽  
Author(s):  
F. Qeusada ◽  
J. Dorado ◽  
D. Acha ◽  
I. Ortiz ◽  
M. Urbano ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Final Concentration ◽  
Computer Assisted ◽  
Preliminary Results ◽  
Cool Storage ◽  
Head Displacement ◽  
Straight Line ◽  
Negative Effect ◽  
Lateral Head ◽  
Motility Parameters

Several studies on sperm cooling and cryopreservation have been done in horses; however, only a few them have been developed in donkeys. In addition, no studies have been performed to freeze cooled stored donkey semen. Therefore, the aim of this study was to determine if it is possible to freeze donkey sperm after 24 h of cool storage. Semen was collected from 4 Andalusian donkeys by artificial vagina. After collection, each sample was separated into 2 aliquots; one of them was immediately frozen (t0) and the other one was cooled and stored before freezing (t24). The cryopreservation procedure consisted of a previous dilution of semen with EquiPro™. After that, semen was centrifuged and the sperm pellet resuspended with Gent® extender plus ethylene glycol (4%) to achieve a final concentration of 100 × 106 sperm mL–1. Sperm was slowly cooled to 5°C, loaded in 0.5-mL plastic straws and frozen in LN vapours. The second aliquot (t24) was diluted with Gent® extender to a final concentration of 50 × 106 sperm mL–1 and then cooled and stored at 5°C for 24 h. After that, cooled semen samples were cryopreserved following the same procedure as described above. Straws were thawed in a water bath at 37° for 30 s. Computer-assisted sperm motility analysis was performed. Total motility (TM), progressive motility (PM), and the following kinematic parameters: velocity curvilinear (VCL; µm s–1), velocity straight line (VSL; µm s–1), velocity average path (VAP; µm s–1), linearity (LIN; %), straightness (STR; %), wobble (WOB; %), amplitude of lateral head displacement (ALH; µm), and beat cross frequency (BCF; Hz) were compared between treatments by ANOVA. Results were expressed as mean ± standard error. Significant differences (P < 0.05) were found between treatments (t0 v. t24) for TM (63.76 ± 4.75 v. 51.67 ± 3.69), PM (36.01 ± 3.19 v. 27.24 ± 2.72), VCL (77.29 ± 0.65 v. 67.56 ± 0.78), VSL (58.50 ± 0.61 v. 52.11 ± 0.76), VAP (67.82 ± 0.64 v. 59.41 ± 0.79), LIN (57.90 ± 0.33 v. 59.53 ± 0.32), STR (70.39 ± 0.30 v. 72.43 ± 0.41), WOB (75.64 ± 0.22 v. 75.48 ± 0.32), ALH (1.88 ± 0.09 v. 1.69 ± 0.10), and BCF (6.28 ± 0.04 v. 6.51 ± 0.06). These preliminary results showed significant differences between cryopreservation at 0 and 24 h post-cooling; however, understanding that direct freezing is better in terms of sperm motility, cryopreservation of cooled stored semen could still be considered good according to the values obtained for sperm motility parameters after thawing. In our opinion, sperm centrifugation before cooling probably improve the results of cryopreservation 24 h post-cooling, due to the negative effect of seminal plasma on sperm viability during storage. In addition, the analysis of other sperm parameters would be useful to check more accurately differences between treatments. In conclusion, sperm motility parameters were higher in donkey semen samples immediately frozen after collection in comparison to semen samples cryopreserved after 24 h of cooling storage. Further studies are needed to improve cooling and cryopreservation procedures for freezing cooled stored donkey semen.

501. BACTERIOSPERMIA AND ITS CONTROL IN BUBALINE SEMEN

2009 ◽  
Vol 21 (9) ◽  
pp. 102
Author(s):  
S. M. H. Andrabi ◽  
M. Shahab
Keyword(s):  
Bacterial Species ◽  
Membrane Integrity ◽  
Negative Control ◽  
Computer Assisted ◽  
Fertility Rates ◽  
Head Displacement ◽  
Straight Line ◽  
Frozen Semen ◽  
Lateral Head

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

scholarly journals Motion Characteristics and Kinematics of Fresh Spermatozoa of Gir, Surti and Murrah Bulls Assessed By Computer Assisted Semen Analyzer

2018 ◽  
Vol 14 (3) ◽  
Author(s):  
A.J. Dhami ◽  
P.K. Pathak ◽  
D.V. Chaudhari ◽  
K. K. Hadiya
Keyword(s):  
Computer Assisted ◽  
Species Variation ◽  
Sperm Velocity ◽  
Mean Values ◽  
Head Displacement ◽  
Straight Line ◽  
Buffalo Semen ◽  
The Mean ◽  
Motion Characteristics ◽  
Lateral Head

A study was carried out on semen of nine breeding bulls, three each of Gir, Surti and Murrah breeds, to evaluate the comparative motion characteristics and kinematics of their fresh spermatozoa by CASA. The ejaculates (n=72, 24 of each breed) having >75% initial motility were diluted @ 80 million sperm/ml using TFYG extender and were assessed for motion characteristics by CASA. The overall mean values of rapid motile and immotile sperm per cent were observed significantly greater in Gir bulls semen, while total motile and slow motile sperms were apparently higher in buffalo semen. The mean values of sperm velocity/ kinematic parameters observed based on all motile sperms in Gir, Surti and Murrah bulls semen were: average path velocity 50.01±1.25, 48.51±1.03 and 49.14±1.30 μm/s; curvilinear velocity 88.62±1.66, 87.90 ±1.74 amd 88.93±1.69 μm/s, straight line velocity 44.51±1.35, 43.14±1.12 amd 41.73±2.24 μm/s; linearity 50.06±1.42, 48.86±1.32 amd 48.49±1.84 %; straightness 85.17±0.92, 84.97±0.88 and 83.90±1.17 %; wobbling index 57.00±1.17, 55.32±1.05 and 55.30±1.48 %; beat-cross frequency 15.55±0.58, 16.14±0.43 and 14.97±0.54 hz; amplitude of lateral head displacement 2.39±0.18, 2.57±0.12 and 2.31±0.14 μm; dancing frequency 208.34±15.52,225.00 ±10.74 and 211.29±13.03 μm2/s, and dancing mean 5.70±0.46, 6.33±0.35 and 6.33±0.50 μm2/s, respectively. Almost similar trend with little higher values were noted for sperm velocity/kinematics based on only progressively motile sperm, without breed/species variation. The semen of all three breeds behaved identically for sperm kinematics. The bull variation was insignificant for all the traits in all the three breeds.

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.

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.

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