Cryobank of Mediterranean Brown Trout Semen: Evaluation of the Use of Frozen Semen up to Six Hours Post-Collection

The aim of this study was to evaluate the effects of different cold-storage time intervals between collection and semen-freezing on both fresh and cryopreserved semen motility parameters and the post-thaw fertilizing ability of Mediterranean brown trout semen. The ejaculates were split into six aliquots and stored on ice from 1 to 6 h, until freezing. Fresh and post-thaw sperm motility was evaluated by a Computer-Assisted Sperm Analysis system, whilst the fertilizing ability was assessed by in vivo trials. In fresh semen, at 3 h of storage, a significant decrease of total motility, linear movement (STR, LIN) and beat cross frequency (BCF) was recorded, whilst the amplitude of lateral displacement of the spermatozoon head (ALH) underwent a significant increase. In frozen semen, no significant difference was observed for all the motility parameters evaluated, except for the total motility between 1 and 6 h of storage and the duration of sperm movement between 1 and 5 h. Cold-storage time did not significantly affect the percentage of live embryos following the use of frozen semen. In conclusion, our results showed that, if necessary, the Mediterranean brown trout semen can be frozen even until 6 h post-collection without losing its fertilizing ability.

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Could the Storage Time Post-collection Affect the Post-thaw Motility and Fertilizing Ability of Mediterranean Brown Trout Semen?

10.21203/rs.3.rs-442986/v1 ◽

2021 ◽

Author(s):

Giusy Rusco ◽

Michele Di Iorio ◽

Roberta Iampietro ◽

Alessandra Roncarati ◽

Stefano Esposito ◽

...

Keyword(s):

Sperm Motility ◽

Brown Trout ◽

Storage Time ◽

Lateral Displacement ◽

Computer Assisted ◽

Cool Storage ◽

Fertilizing Ability ◽

Linear Movement ◽

Motility Parameters

Abstract The aim of this study was to evaluate the effect of different cool storage time intervals between collection and semen freezing on both fresh and cryopreserved semen motility parameters and the post-thaw fertilizing ability of Mediterranean brown trout semen. The ejaculates were split into six aliquots and stored on ice for 1 to 6 hours, until freezing. Fresh and post-thawing sperm motility were evaluated by Computer-Assisted Sperm Analysis system, whilst the fertilizing ability was assessed by in vivo trials. In fresh semen, at 3 h of storage, a significant decrease of total motility, linear movement (STR, LIN) and beat cross frequency was recorded, whilst the amplitude of lateral displacement of the spermatozoon head underwent a significant increase. Velocity parameters (VCL, VAP and VSL) were not affected by the cold storage time, whilst the duration of sperm movement was significantly higher at 1h compared to the other times tested. Freezing procedure overall decreased almost all post-thaw sperm motility parameters, however no significant differences was observed over time, both in term of fast and linear movement. Cool storage time did not significantly affect the percentage of post-thaw eyed embryos. Our results showed that Mediterranean brown trout semen can be stored on ice even up to 6 hours before freezing, without decreasing its post-thawing quality and fertilizing ability.

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67 EFFECTS OF ANTIOXIDANTS LACTOFERRIN AND CATALASE ON STALLION FROZEN SEMEN

Reproduction Fertility and Development ◽

10.1071/rdv27n1ab67 ◽

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.

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24 BUFFALO (BUBALUS BUBALIS) SOMATIC CELL NUCLEAR TRANSFER EMBRYOS PRODUCED FROM FROZEN–THAWED SEMEN-DERIVED SOMATIC CELLS: EFFECT OF TRICHOSTATIN A ON THE IN VITRO AND IN VIVO DEVELOPMENTAL POTENTIAL, QUALITY, AND EPIGENETIC STATUS

Reproduction Fertility and Development ◽

10.1071/rdv27n1ab24 ◽

2015 ◽

Vol 27 (1) ◽

pp. 104

Author(s):

N. L. Selokar ◽

M. Saini ◽

H. Agrawal ◽

P. Palta ◽

M. S. Chauhan ◽

...

Keyword(s):

Somatic Cell ◽

Somatic Cell Nuclear Transfer ◽

Nuclear Transfer ◽

Trichostatin A ◽

Somatic Cells ◽

Developmental Potential ◽

Reconstructed Embryos ◽

Frozen Semen ◽

Significant Difference

Cryopreservation of semen allows preservation of somatic cells, which can be used for the production of progeny through somatic cell nuclear transfer (SCNT). This approach could enable restoration of valuable high-genetic-merit progeny-tested bulls, which may be dead but the cryopreserved semen is available. We have successfully produced a live buffalo calf by SCNT using somatic cells isolated from >10 year old frozen semen (Selokar et al. 2014 PLoS One 9, e90755). However, the calf survived only for 12 h, which indicates faulty reprogramming of these cells. The present study was, therefore, carried out to study the effect of treatment with trichostatin A (TSA), an epigenetic modifier, on reprogramming of these cells. Production of cloned embryos and determination of quality and level of epigenetic markers in blastocysts were performed according to the methods described previously (Selokar et al. 2014 PLoS One 9, e90755). To examine the effects of TSA (0, 50, and 75 nM), 10 separate experiments were performed on 125, 175, and 207 reconstructed embryos, respectively. The percentage data were analysed using SYSTAT 12.0 (SPSS Inc., Chicago, IL, USA) after arcsine transformation. Differences between means were analysed by one-way ANOVA followed by Fisher's least significant difference test for significance at P < 0.05. When the reconstructed buffalo embryos produced by hand-made clones were treated with 0, 50, or 75 nM TSA post-electrofusion for 10 h, the cleavage percentage (100.0 ± 0, 94.5 ± 2.3, and 96.1 ± 1.2, respectively) and blastocyst percentage (50.6 ± 2.3, 48.4 ± 2.7, and 48.1 ± 2.6, respectively), total cell number (274.9 ± 17.4, 289.1 ± 30.1, and 317.0 ± 24.2, respectively), and apoptotic index (3.4 ± 0.9, 4.5 ± 1.4, and 5.6 ± 0.7, respectively) in Day 8 blastocysts were not significantly different among different groups. The TSA treatment increased (P < 0.05) the global level of H4K5ac but not that of H3K18a in embryos treated with 50 or 75 nM TSA compared with that in controls. In contrast, the level of H3K27me3 was significantly lower (P < 0.05) in cloned embryos treated with 75 nM TSA than in embryos treated with 50 nM TSA or controls. The ultimate test of the reprogramming potential of any donor cell type is its ability to produce live offspring. To examine the in vivo developmental potential of the 0, 50, or 75 nM TSA treated embryos, we transferred Day 8 blastocysts, 2 each to 5, 6, and 5 recipients, respectively, which resulted in 2 pregnancies from 75 nM TSA treated embryos. However, one pregnancy was aborted in the first trimester and the other in the third trimester. In conclusion, TSA treatment of reconstructed embryos produced from semen-derived somatic cells alters their epigenetic status but does not improve the live birth rate. We are currently optimizing an effective strategy to improve the cloning efficiency of semen-derived somatic cells.

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3D culture model and computer-assisted videomicroscopy to analyze migratory behavior of noninvasive and invasive bronchial epithelial cells

AJP Cell Physiology ◽

10.1152/ajpcell.00319.2005 ◽

2005 ◽

Vol 289 (6) ◽

pp. C1547-C1552 ◽

Cited By ~ 19

Author(s):

Salma Hazgui ◽

Noël Bonnet ◽

Jérôme Cutrona ◽

Béatrice Nawrocki-Raby ◽

Myriam Polette ◽

...

Keyword(s):

Cell Migration ◽

Cell Line ◽

3D Model ◽

Migratory Behavior ◽

Computer Assisted ◽

Data Set ◽

Significant Difference ◽

Kinetics Of ◽

Invasive Cell

To date, most of the studies in the field of cell migration have been applied to two-dimensional (2D) models. To mimic the three-dimensional (3D) conditions similar to those observed in vivo during tumor invasion, we developed a 3D model of cell migration in which cells were embedded in a collagen I matrix placed in a double-compartment chamber. Using time-lapse videomicroscopy and interactive cell tracking in a four-dimensional data set, we determined the cell trajectories and their migration kinetics. We compared the 2D and 3D migratory behavior of a noninvasive cell line (16HBE) with the migratory behavior of an invasive cell line (BZR). Our results show that the 3D migration kinetics of the noninvasive cell line were lower than the migration kinetics of the invasive cell line. In contrast, in 2D models, no significant difference was observed between the two cell lines. To validate our 3D model, we further investigated the effect of epidermal growth factor (EGF), a promoter of tumor cell motility and invasion on the noninvasive cell line (16HBE). EGF increased significantly the migration kinetics of the noninvasive cell line. Our results show that the 3D model of cell migration allowed us to differentiate the migratory behavior of invasive and noninvasive cells and that such a model can help in the development of molecular targeted therapy as it approaches the in vivo conditions.

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Effect of cryopreservation on sperm motility parameters and fertilizing ability of brown trout semen

Aquaculture ◽

10.1016/j.aquaculture.2014.05.037 ◽

2014 ◽

Vol 433 ◽

pp. 62-65 ◽

Cited By ~ 46

Author(s):

J. Nynca ◽

G.J. Dietrich ◽

S. Dobosz ◽

J. Grudniewska ◽

A. Ciereszko

Keyword(s):

Sperm Motility ◽

Brown Trout ◽

Fertilizing Ability ◽

Motility Parameters

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Capacitation-Induced Mitochondrial Activity Is Required for Sperm Fertilizing Ability in Mice by Modulating Hyperactivation

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.767161 ◽

2021 ◽

Vol 9 ◽

Author(s):

María Milagros Giaccagli ◽

Matías Daniel Gómez-Elías ◽

Jael Dafne Herzfeld ◽

Clara Isabel Marín-Briggiler ◽

Patricia Sara Cuasnicú ◽

...

Keyword(s):

Mitochondrial Activity ◽

Computer Assisted ◽

Specific Staining ◽

Sperm Analysis ◽

Functional Changes ◽

Fertilizing Ability ◽

Sperm Fertilizing Ability

To become fully competent to fertilize an egg, mammalian sperm undergo a series of functional changes within the female tract, known as capacitation, that require an adequate supply and management of energy. However, the contribution of each ATP generating pathway to sustain the capacitation-associated changes remains unclear. Based on this, we investigated the role of mitochondrial activity in the acquisition of sperm fertilizing ability during capacitation in mice. For this purpose, the dynamics of the mitochondrial membrane potential (MMP) was studied by flow cytometry with the probe tetramethylrhodamine ethyl ester (TMRE). We observed a time-dependent increase in MMP only in capacitated sperm as well as a specific staining with the probe in the flagellar region where mitochondria are confined. The MMP rise was prevented when sperm were exposed to the mitochondrial uncoupler carbonyl cyanide m-chlorophenyl hydrazine (CCCP) or the protein kinase A (PKA) inhibitor H89 during capacitation, indicating that MMP increase is dependent on capacitation and H89-sensitive events. Results showed that whereas nearly all motile sperm were TMRE positive, immotile cells were mostly TMRE negative, supporting an association between high MMP and sperm motility. Furthermore, CCCP treatment during capacitation did not affect PKA substrate and tyrosine phosphorylations but produced a decrease in hyperactivation measured by computer assisted sperm analysis (CASA), similar to that observed after H89 exposure. In addition, CCCP inhibited the in vitro sperm fertilizing ability without affecting cumulus penetration and gamete fusion, indicating that the hyperactivation supported by mitochondrial function is needed mainly for zona pellucida penetration. Finally, complementary in vivo fertilization experiments further demonstrated the fundamental role of mitochondrial activity for sperm function. Altogether, our results show the physiological relevance of mitochondrial functionality for sperm fertilization competence.

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

Reproduction Fertility and Development ◽

10.1071/rdv32n2ab121 ◽

2020 ◽

Vol 32 (2) ◽

pp. 187

Author(s):

M. Felix ◽

I. Ortiz ◽

H. Resende ◽

J. Brom-de-Luna ◽

C. Love ◽

...

Keyword(s):

Sperm Motility ◽

Sperm Quality ◽

Phosphodiesterase Inhibitor ◽

Petri Dish ◽

Poor Quality ◽

Computer Assisted ◽

Progressive Motility ◽

Frozen Semen ◽

Significant Difference ◽

The Right

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

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15 QUANTIFICATION OF BULL SPERM TRAITS AS ASSESSED BY COMPUTER-ASSISTED SEMEN ANALYSIS AND THE RELATIONSHIP TO PREGNANCY RATE FOLLOWING CONTROLLED BREEDING

Reproduction Fertility and Development ◽

10.1071/rdv29n1ab15 ◽

2017 ◽

Vol 29 (1) ◽

pp. 115

Author(s):

M. L. Mphaphathi ◽

M. M. Seshoka ◽

F. V. Ramukhithi ◽

Z. C. Raphalalani ◽

T. R. Netshirovha ◽

...

Keyword(s):

In Vitro Fertilization ◽

Pregnancy Rate ◽

Semen Analysis ◽

Computer Assisted ◽

Vitro Fertilization ◽

Significant Difference ◽

Bull Sperm ◽

Sperm Traits

The bull’s contribution through artificial insemination to reproductive efficiency is of great biological importance. The objectives were (1) to compare the oestrous synchronization response of Bonsmara and Nguni cows; and (2) to find the relationship between cow’s conception rate (in vivo and in vitro fertilization) and bull sperm motility rate assessed by computer-assisted semen analysis (CASA) following AI. For the in vivo sperm fertility test, 100 Bonsmara and 482 Nguni cows were randomly selected and subjected to oestrous synchronization protocol and AI with frozen–thawed assessed semen by CASA before AI. Briefly at Day 0, cows were inserted with an intravaginal CIDR® (1.9 g), which was removed on Day 7. Prostaglandin was then administered (2 mL) on Day 8 and a heatmount detector was placed on the hindquarter of each cow. For the in vitro sperm fertility test, collected oocytes from slaughterhouse were in vitro matured (n = 360) and in vitro fertilized (sperm/mL) in 100-µL droplets (final volume) of BO-IVF medium per treatment bulls (Bonsmara or Nguni bull). The frozen/thawed semen straws of Bonsmara and Nguni bulls were randomly selected and used under the same IVF conditions. The thawed bull’s sperm characteristics were examined by CASA before in vitro fertilization. Data were analysed using ANOVA. Treatment means were compared using the Fisher’s protected least significant difference t-test. There was no significant difference in oestrous response for the Bonsmara (83.0%) and Nguni (90.8%) cows, respectively. The Bonsmara cows recorded a significantly higher pregnancy rate (59.0%) compared with the Nguni (37.1%) cows (P < 0.05). Sperm traits such as total motility (TM), progressive motility and rapid were found to be positively correlated with conception rate (r = 0.06, 0.03, and 0.08, respectively; P < 0.01), although correlations were low. There was no difference in the average frozen–thawed sperm TM rate of Nguni (92.2%) and Bonsmara (81.0%). There was a lower fertilization rate following IVF with Bonsmara and Nguni bull sperm. In conclusion, Nguni cows had similar oestrous response as Bonsmara cows. The sperm traits from Bonsmara and Nguni bulls were found to be related to in vivo conception and in vitro fertilization rate when sperm cells were assessed by CASA technology. However, the pregnancy rate was lower in Nguni cows.

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

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab84 ◽

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

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Computer assisted sperm analysis: Relationship between the movement characteristics of buffalo spermatozoa and sire fertility

Indian Journal of Animal Research ◽

10.18805/ijar.10768 ◽

2016 ◽

Author(s):

Raushan K Singh ◽

A. Kumaresan ◽

M. A. Mir ◽

P. Kumar ◽

S. Chhillar ◽

...

Keyword(s):

Computer Assisted ◽

Progeny Testing ◽

Kinematic Parameters ◽

Murrah Buffalo ◽

Sperm Analysis ◽

Head Displacement ◽

Movement Characteristics ◽

Frozen Semen ◽

Lateral Head

The present study was undertaken to identify the differences in sperm kinematics between buffalo bulls with different fertility ratings. Murrah buffalo bulls (n=9) that were routinely used for breeding purpose under progeny testing programme were utilized for the study. Bull fertility was determined based on in vivo fertility trials and the conception rates (CR) were adjusted for different non-genetic parameters. Based on the adjusted CR, bulls were classified into high, medium and low fertile group. Frozen semen samples of these bulls were obtained and sperm kinematic parameters were assessed using a computer assisted sperm analyzer. The kinematic parameters analyzed included the curvilinear velocity (VCL), the linear velocity (VSL), the average path velocity (VAP), the amplitude of lateral head displacement (ALH), the linearity (LIN), the straightness coefficient (STR) and the beat cross frequency (BCF). In high fertile bulls, the proportion of motile spermatozoa was higher (p<0.001) than the medium and low fertile bulls. The VAP and VCL of sperm motion were significantly higher (P<0.05) in high fertile bulls compared to either medium or low fertile bulls. The VSL was significantly lower in low fertile bulls (P<0.005) compared to either high or medium fertile bulls. Spermatozoa from high fertile bulls had significantly higher (P<0.05) BCF, STR, ALH and LIN compared to either medium or low fertile bulls. Buffalo bull fertility was significantly and positively correlated with sperm motility, VAP, VSL, VCL and ALH.

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