Effect of Bioxcell® and Triladyl® Extenders and Removal of Seminal Plasma on Equilibrated and Cryopreserved Semen from South African Unimproved Indigenous Bucks

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Nethenzheni LP ◽  
◽  
Mphaphathi ML ◽  
Madzhie LR ◽  
Negota NC ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Membrane Integrity ◽  
The Other ◽  
Semen Parameters ◽  
Lower Percentage ◽  
Mean Values ◽  
Sperm Analysis ◽  
Progressive Motility ◽  
Frozen Semen ◽  
Motility Rate

The objectives of the study were to evaluate the effect of two extenders (Triladyl® and Bioxcell®) and the removal of seminal plasma on indigenous buck’s semen. Semen was collected from six indigenous bucks using an electro-ejaculator. Raw semen was pooled and randomly allocated into six groups as follows: (i) Raw non-washed, (ii) Raw washed, (iii) Triladyl®-washed, (iv) Triladyl®-non-washed, (v) Bioxcell®-washed and (vi) Bioxcell®-non-washed. Both the Triladyl® and Bioxcell® washed semen samples groups were diluted (1:4 v/v) with Phosphate Buffered Saline (PBS) then centrifuged at 1500x g for ten min and seminal plasma was removed. The groups were analysed for spermatozoa motility rates using Computer-Aided Sperm Analysis (CASA). The spermatozoa viability was assessed using Eosin-Nigrosin, acrosome integrity using Spermac, chromatin structure using Acridine Orange, and mitochondria using JC-1 staining solutions. Semen samples were diluted (1:4 v/v) as follows: Triladyl® (washed and non-washed) or Bioxcell® (washed and non-washed) and then equilibrated at 5°C for 2 hours. Equilibrated semen samples in 0.25 mL French straws were placed 5 cm above a Liquid Nitrogen (LN2) vapour for 10 min, and stored for one month. Frozen semen straws per treatment group were thawed at 37°C for 30 seconds. Significant differences among the mean values of semen parameters were determined by Tukey’s test using ANOVA, GLM procedure of SAS version 12.1 of 2010. The spermatozoa progressive motility rate in non-washed semen extended with Bioxcell® was significantly higher (89.6±7.5a) compared with that of non-washed Triladyl®, washed Bioxcell® and Triladyl® (P<0.05). Live spermatozoa percentage in washed semen extended with Triladyl® extender was reduced (27.7±17.1) significantly compared with the other groups (P<0.05). There was a lower percentage of spermatozoa with high mitochondrial membrane potential in non-washed and washed semen extended with Bioxcell® (39.5±23.2 and 37.9±28.6, respectively) compared with that of non-washed and washed semen extended with Triladyl® (P>0.05). The spermatozoa progressive motility rate in non-washed semen extended with Bioxcell® (58.5±10.0) extender was significantly higher compared with that of the other groups (P<0.05). There was a higher live and normal spermatozoa percentage in non-washed semen extended with Bioxcell® (45.7±21.2) compared with that of the other groups (P<0.05). In conclusion, Washing of seminal plasma in semen extended with Triladyl® was not essential, as it lowered viability, progressive motility and chromatin membrane integrity prior and post-cryopreservation. However, Bioxcell® extender was found to be more suitable for preserving spermatozoa during equilibration and freezing/thawing process of non-washed and washed buck semen.

46 Effect of Bioxcell® and Triladyl® Extenders on Washed Semen of South African Indigenous Bucks

2018 ◽  
Vol 30 (1) ◽  
pp. 162
Author(s):  
L. P. Nethenzheni ◽  
M. L. Mphaphathi ◽  
N. C. Negota ◽  
T. L. Nedambale
Keyword(s):  
Treatment Group ◽  
South African ◽  
Liquid Nitrogen ◽  
Seminal Plasma ◽  
Semen Parameters ◽  
Mean Values ◽  
Treatment Groups ◽  
Frozen Semen ◽  
The Mean

Semen extenders and seminal plasma are vital for cryopreservation of buck semen. The objectives of the study were to evaluate the effect of 2 extenders: Triladyl® (Minitube, Tiefenbach, Bavaria) and Bioxcell® (IMV, L’Aigle, France) and the removal of seminal plasma on buck semen. Six indigenous bucks were used in this study and 6 ejaculates were collected from individual bucks. The semen was pooled and then randomly allocated into 6 groups: (1) raw-washed, (2) raw-non-washed, (3) Triladyl®-washed, 4) Triladyl®-non-washed, (5) Bioxcell®-washed, and (6) Bioxcell®-non-washed. Spermatozoa viability was assessed using Eosin-Nigrosin and morphology using Spermac® (Vitrolife, Göteborg, Sweden) stains. The washed semen samples were all diluted into (1:4 v/v) with PBS and centrifuged at 1500 × g for 10 min. Semen samples were then extended with Triladyl® or Bioxcell® per treatment groups and equilibrated for 2 h at 5°C. The semen samples were loaded into straws per treatment groups and placed 5 cm above a liquid nitrogen vapour for 10 min and then stored at –196°C until use. After 1 month of storage, frozen semen straws per treatment group were thawed at 37°C for 30 s, and spermatozoa parameters were analysed post-thaw. Significant differences among the mean values of semen parameters were determined by Tukey’s test using ANOVA, GLM procedure of SAS version 12.1 of 2010 (SAS Institute Inc., Cary, NC, USA). There was a higher (P < 0.05) live and normal spermatozoa percentage in non-washed semen extended with Bioxcell® (45.7 ± 21.2) than the semen extended with Triladyl® (24.5 ± 22.2%). Live and normal spermatozoa percentages were drastically reduced in the Bioxcell® (5.2 ± 4.9) and Triladyl® (6.9 ± 8.6%) washed semen groups. There was a higher (P < 0.05) percentage of spermatozoa with head abnormalities in non-washed semen extended with Triladyl® (20.4 ± 10.2), compared with the semen extended with Bioxcell® (18.3 ± 12.4%) following freeze-thawing. There was a higher (P < 0.05) percentage of spermatozoa with head abnormalities in washed semen samples extended with Triladyl® (34.0 ± 16.0) compared with the semen extended with Bioxcell® (10.1 ± 7.0%). There were higher (P < 0.05) percentages of spermatozoa with coiled tail abnormalities in washed semen extended with Bioxcell® (65.4 ± 25.0) compared with Triladyl® (35.9 ± 21.6%). In conclusion, the liveability of spermatozoa was negatively affected by washing of semen extended with Bioxcell® and Triladyl® extender. Bioxcell® significantly increased tail abnormalities and Triladyl® gave less protection against head abnormalities following cryopreservation of South African unimproved indigenous bucks’ semen.

65 ARE “BAD FREEZER” STALLIONS ALSO “BAD COOLER” STALLIONS?

2015 ◽  
Vol 27 (1) ◽  
pp. 125
Author(s):  
C. Ramires Neto ◽  
M. M. B. Castro-Chaves ◽  
Y. F. R. Sancler-Silva ◽  
R. C. Uliani ◽  
P. V. L. Oliveira ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Seminal Plasma ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Cooling Process ◽  
Freezing Resistance ◽  
Plasma Membrane Integrity ◽  
Progressive Motility ◽  
Frozen Semen ◽  
Fresh Semen

Several factors can interfere with sperm cryopreservation resistance, especially the genetic factors and those related to the plasma membrane composition of the sperm and seminal plasma. However, it is still unclear if the same factors that confer freezing resistance will perform the same role during the cooling process. Thus, the aim of this study was to determine the relation between the resistance to freezing and cooling processes in stallions. Two ejaculates from each of 75 stallions were used. All animals showed good quality of fresh semen (total motility higher than 60% and plasma membrane integrity higher than 50%). After collection, the semen was diluted 1 : 1 with commercial skim milk-based extender (Botu-SemenTM, Botupharma, Brazil) and then a part was designed to cooling and the another to freezing. The cooled semen was divided into 2 groups: Group PS, in which the semen was diluted with Botu-SemenTM at a concentration of 50 × 106 sperm mL–1, and Group SPS, which was subjected to a centrifugation at 600 × g for 10 min and resuspended with Botu-SemenTM at 50 × 106 sperm mL–1. Semen samples from both groups were placed in the same cooling passive system for a period of 24 h/5°C. To accomplish the freezing process, the semen sample was subjected to centrifugation at 600 × g for 10 min. The supernatant was discarded, and the pellet was re-suspended in a Botu-CrioTM. The straws were frozen according to the manufacture. The sperm parameters from fresh semen, cooled semen for 24 h with and without seminal plasma, and frozen semen were evaluated for kinetics by computer-assisted semen analysis and for plasma membrane integrity (IMP%) by epi-fluorescence microscopy. The animals were classified in relation to their resistance to cooling and freezing processes as follow: “bad coolers” – reduction in sperm total motility and in plasma membrane integrity higher than 35% after 24 h of cooling in samples with seminal plasma; “good coolers” – reduction in sperm total motility and in plasma membrane integrity lower than 35% after 24 h of cooling in samples with seminal plasma; “bad freezer” – sperm total motility lower than 40% and progressive motility lower than 20% in seminal sample after thawing; “good freezer” – sperm total motility higher than 60% and progressive motility higher than 30% in seminal sample after thawing. The comparison between the resistance to cooling and freezing processes was performed by Fisher's exact test. The level of significance was 5%. No difference (P < 0.05) between the resistance to cooling and freezing processes was observed. The percentage of stallions “good freezer” and “good cooler” was 54%, “good freezer” and “bad cooler” was 22.6%, “bad freezer” and “good cooler” was 12%, and “bad freezer” and “bad cooler” was 10.6%. Within stallions classified as “good freezer” and “bad cooler,” 52.9% also were “good cooler” when the seminal plasma was removed before the cooling process, and 47.1% remained as “bad cooler.” The result of this study demonstrates that there is a strong relation between the resistance to cooling and freezing processes in stallions. In stallions categorized as “bad cooler,” the seminal plasma presents a major influence on the quality and longevity of cooled semen.

240 FUNCTIONAL TRAITS OF CAT SPERM DURING DISTINCT MATURATION STATUS

2011 ◽  
Vol 23 (1) ◽  
pp. 218
Author(s):  
E. G. A. Perez ◽  
M. Nichi ◽  
C. A. Baptista Sobrinho ◽  
P. A. A. Góes ◽  
A. Dalmazzo ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Lipid Peroxidation ◽  
Structural Damage ◽  
Membrane Integrity ◽  
Domestic Cat ◽  
Lower Percentage ◽  
Mitochondrial Activity ◽  
Computer Assisted ◽  
Suitable Model ◽  
Sperm Analysis

Sperm recovery from the caudae epididymides can be advantageous for preserving semen of endangered animal species. In this context, the domestic cat is a suitable model for the study of sperm physiology in endangered feline species and the research on epididymal sperm preservation combined with the use of reproductive biotechnologies including intracytoplasmic sperm injection (ICSI). The aim of the present study was to examine the sperm collected from the cauda and caput of the cat epididymis using functional tests. Testicles and epididymides from 5 adult tomcats were collected by orchiectomy and maintained at 4°C for 4 h, until semen collection. Semen samples were collected from the epididymal tail and head by careful dissection. Samples were then analysed for motility by computer assisted sperm analysis (CASA; only for the caudal sperm). The 3-3′ diaminobenzidine stain was used as an index of mitochondrial activity, the eosin nigrosin stain as an index of membrane integrity, the simple stain (fast green/Bengal rose) as an index of acrosome integrity, and the measurement of thiobarbituric acid reactive substances (TBARS) as an index of lipid peroxidation. Statistical analysis was performed using the SAS System for Windows (SAS Institute Inc., Cary, NC, USA; least significant differences test and Spearman correlation; P < 0.05). No motility was observed in samples collected from the epididymal head, whereas samples from the tail showed 50.0 ± 4.2% motile spermatozoa. Surprisingly, more spermatozoa with high mitochondrial activity were found in the epididymal head than in samples from the tail (74.0 ± 3.5 v. 50.0 ± 4.3%, respectively). Similarly, samples collected from the head showed a higher susceptibility against the attack of ROS (31.9 ± 5.5 v. 16.3 ± 7.1 ng of TBARS/106 sperm, respectively). Furthermore, epididymal head sperm showed a lower percentage of sperm with intact membrane and a higher percentage of sperm with intact acrosome (44.9 ± 3.3 and 78.4 ± 1.8 v. 66.4 ± 4.2 and 56.7 ± 4.4%, respectively). Our results demonstrate that, during maturation, feline sperm are subjected to high oxidative stress, as shown by the lipid peroxidation assay, which would lead to structural damage to biomolecules, DNA, lipids, carbohydrates and proteins, as well as other cellular components, such as mitochondria, and acrosomal impairment. Similar results were found in humans, in which higher levels of oxidative stress occurred in the post-testicular environment. The plasma membrane seems to be more resistant to damages. This may be due to the described rearrangement in the lipid profile occurring during maturation, but studies to test this hypothesis are still underway.

69 REACTIVE OXYGEN SPECIES EVALUATION OF DONKEY FROZEN SEMEN ADDED TO HOMOLOGOUS SEMINAL PLASMA ON POST-THAW

2015 ◽  
Vol 27 (1) ◽  
pp. 127
Author(s):  
P. V. L. Oliveira ◽  
J. V. Oliveira ◽  
C. Ramires Neto ◽  
Y. F. R. Sancler-Silva ◽  
C. P. Freitas-Dell'aqua ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Kinetic Parameters ◽  
Seminal Plasma ◽  
Membrane Integrity ◽  
Reactive Oxygen ◽  
Computer Assisted ◽  
Oxygen Species ◽  
Inflammation Response ◽  
Frozen Semen ◽  
Uterine Inflammation

For many years the pregnancy rate of donkey frozen semen presented lower results in donkey jennies; however, a recent study showed an increase in pregnancy rates using frozen semen added to seminal plasma on post-thaw. A hypothesis for this result is the higher uterine inflammation response after breeding when using seminal plasma. The same studies demonstrated higher uterine inflammation in the presence of higher reactive oxygen species concentration. The aim of the present study was to evaluate the content of reactive oxygen species in donkey frozen semen added to homologous seminal plasma on post-thaw. Five ejaculates from each 3 donkeys were used. Semen was diluted (1 : 1) with a skim milk-based extender (Botu-SemenTM, Botupharma, Brazil). The semen was frozen with Botu-CryoTM extender (Botupharma, Brazil) in an isothermal box in straws containing 100 × 106 of total sperm. The samples were thawed at 46°C for 20 s. After this, the straws of each donkey were divided in 2 group: control group (CG), in which the semen was incubated at 37°C for 5 min, and plasma seminal group (PG), in which the semen was incubated at 37°C for 5 min with 70% of homologous seminal plasma. Sperm kinetic parameters were evaluated by computer-assisted semen analysis, and the plasma membrane integrity (propidium iodide and fluorescein isothiocyanate -PSA) and reactive oxygen species (5–6-carboxi-2,7-diclorodihidrofluoresceindiacetate) were evaluated by flow cytometer. Comparison of sperm parameters was performed by t-test. Total motility (%, CG = 75.4 ± 8.2a v. PG = 57.5 ± 16.4b), progressive motility (%, CG = 42.0 ± 8.7a v. PG = 33.3 ± 13.2b), progressive angular velocity (μm/s, CG = 95.8 ± 10.8a v. PG = 88.9 ± 10.9b), and percentage of rapid sperm (%, CG = 58.4 ± 12.5a v. PG = 41.0 ± 17.3b) were higher in CG compare with PG. No difference (P < 0.05) was observed in membrane integrity (%, CG = 20.7 ± 7.4 v. PG = 20.6 ± 7.8); however, reactive oxygen species (%, CG = 12.3 ± 10.6a v. PG = 81.8 ± 32.5b) were higher in PG. The results of this study showed that the addition of homologous seminal plasma on post-thaw decreases the sperm kinetic parameters and viability of donkey frozen semen but increases reactive oxygen species, and this may cause higher uterine inflammation response in donkey jennies and increase their fertility.

267 EFFECT OF SEMINAL PLASMA IN LAMA GLAMA SPERM

2015 ◽  
Vol 27 (1) ◽  
pp. 223 ◽  
Author(s):  
M. Carretero ◽  
F. Fumuso ◽  
M. Miragaya ◽  
C. Herrera ◽  
S. Giuliano
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Membrane Integrity ◽  
South American ◽  
South American Camelids ◽  
Progressive Motility ◽  
Coomassie Blue ◽  
Blue Stain ◽  
Lama Glama ◽  
Motility Of Sperm

In South American camelids, raw semen only presents sperm with oscillatory movements. Therefore, it is necessary to treat these cells to enable them to acquire progressive motility. The effects of raw seminal plasma (SP) on sperm movement patterns (oscillatory, progressive, and hyperactive) have apparently not yet been reported. The objective of this study was to determine effects of raw seminal plasma on sperm motility, viability, and acrosomal status in fresh llama semen. A total of 15 ejaculates were collected (electroejaculation) from 5 llamas (n = 5, r = 3). Each ejaculate was diluted 4 : 1 in 0.1% collagenase in HEPES-TALP (HT) medium and incubated 4 min at 37°C, with the objective of separating spermatozoa from SP. Immediately after incubation, each ejaculate was divided into 2 and centrifuged for 8 min at 800 × g. Pellets were resuspended in either HT or raw SP and maintained at 37°C until evaluation (at 0, 1.5, and 3 h). Sperm motility was evaluated using a phase contrast microscope and a warm stage. Propidium iodide and carboxyfluorescein diacetate were used for assessing membrane integrity (viability). Acrosomal status was evaluated with the Coomassie blue stain. A split-plot design was used with treatment as a factor, with 2 levels (HT and SP) and time as the other factor, with 3 levels (0, 1.5, and 3 h), and blocked by males. There was no significant interaction between treatments (HT and SP) and times (0, 1.5, and 3 h) for each of the seminal characteristics evaluated. Progressive sperm motility was observed after collagenase treatment in all samples. Progressive motility disappeared immediately after the addition of raw SP and showed only oscillatory movements. In contrast, samples incubated in HT maintained progressive motility and became hyperactive. There were no differences (P > 0.05) in total motility of sperm incubated in HT among incubation times (0 h: 30.8 ± 18.9%; 1.5 h: 26.5 ± 11.5%; and 3 h: 21.5 ± 13.5%). However, in samples incubated with SP, a decrease (P < 0.05) in total sperm motility was detected after 3 h of incubation (0 h: 16.5 ± 12.6%, 3 h: 2.3 ± 3.2%). Sperm viability was not different (P > 0.05) between treatments (HT and SP); samples incubated in HT retained 78.4% of the initial viability (32.8/41.8, 3 h/0 h), and samples incubated in SP retained 69.7% of their initial viability (24.4/35.0, 3 h/0 h). The percentage of spermatozoa with intact acrosomes was not different (P > 0.05) between treatments (HT and SP); however, the percentage of sperm with intact acrosomes decreased after 3 h of incubation in both samples (HT and SP). Due to the presence of a high percentage of progressive and hyperactive motile sperm in samples incubated in HT and their absence in samples incubated in SP, we concluded that raw seminal plasma preserved oscillatory sperm motility. Further studies are needed to understand the effects of SP on South American camelid spermatozoa.

20 Assessment of Motion and Kinematic Characteristics of Semen from Four Cattle Breeds Using Computer-Aided Sperm Analysis

2018 ◽  
Vol 30 (1) ◽  
pp. 149
Author(s):  
M. L. Mphaphathi ◽  
M. M. Seshoka ◽  
T. R. Netshirovha ◽  
Z. C. Raphalalani ◽  
N. Bovula ◽  
...  
Keyword(s):  
Objective Evaluation ◽  
Semen Parameters ◽  
Sperm Cells ◽  
Sperm Analysis ◽  
Progressive Motility ◽  
Cattle Breeds ◽  
Bull Semen ◽  
Head Displacement ◽  
Computer Aided ◽  
Sperm Motion

Subjective semen evaluation using standard optical microscopy is the most common practice. Semen parameters routinely assessed are volume, concentration, progressive motility, and morphology. However, computer-aided sperm analysis (CASA) represents an objective evaluation, sperm assessment that are reproducible and reliable. Such semen parameters have not been evaluated in Afrikaner, Brahman, and Bonsmara bulls’ semen. The present study evaluated the sperm motion and kinematics characteristics of semen from stud Afrikaner, Brahman, Bonsmara, and Nguni bulls using CASA technology. The electro-ejaculator was used for semen collection from Afrikaner (n = 11), Brahman (n = 7), Bonsmara (n = 10) and Nguni (n = 16) bulls of known and proven fertility. Semen was collected following 4 days of resting period. The bulls ranged between 5 and 6 years of age. After collection, the semen samples were immediately transferred to a thermo-flask and maintained at 37°C for further evaluation in the mobile laboratory (Nedambale, 2014). The CASA-Sperm Class Analyzer® system (Microptic, Barcelona, Spain) was used to evaluate sperm motion, velocity, and kinematic parameters or characteristics of raw/fresh semen from 4 cattle breeds. Data were analysed using GenStat® statistical programme (VSN International, Hemel Hempstead, United Kingdom). Treatment means were compared using one-way ANOVA. The total sperm motility rate was similar for all breeds: Afrikaner (92.2 ± 4.2), Brahman (90.7 ± 9.0), Bonsmara (93.9 ± 4.0), and Nguni (96.0 ± 2.7). However, Brahman and Afrikaner bull semen had higher sperm cells moving in a progressive motility of 57.3 and 45.6%, respectively, compared with other breeds (P < 0.05). Nguni, Afrikaner, and Bonsmara had the highest sperm cells moving in a rapid movement of 73.7, 72.4, and 67.4% (P > 0.05), respectively. The bulls sperm trajectories had a variation, as they were recorded to be irregular and not linear (P < 0.05). The straight-line sperm velocity (µm s−1), wobbling %, and amplitude of lateral head displacement % was similar for the 4 breeds (P > 0.05). In conclusion, CASA technology was a useful technique for assessing differences in sperm motion and kinematic (motility and velocity characteristics) among different bull breeds.

scholarly journals Comparison of three different media for freezing of epididymal sperm from the African buffalo (Syncerus caffer) and influence of equilibration time on the post-thaw sperm quality

2004 ◽  
Vol 71 (3) ◽  
Author(s):  
F.C. Herold ◽  
K. De Haas ◽  
D. Cooper ◽  
B. Colenbrander ◽  
J.O. Nothling ◽  
...  
Keyword(s):  
South Africa ◽  
Sperm Quality ◽  
The Other ◽  
Semen Parameters ◽  
Equilibration Time ◽  
African Buffalo ◽  
Epididymal Sperm ◽  
Progressive Motility ◽  
Reproductive Techniques ◽  
Buffalo Semen

Assisted reproductive techniques might prove themselves useful tools in producing buffaloes free of specific diseases (BFSD), which are in demand in South Africa. Freezing protocols for African buffalo semen must not only result in good post-thaw qualities, but must also be practical. Epididymal sperm from six mature African buffalo bulls was collected, diluted with three different semen extenders and frozen. Pre-freezing equilibration times between 2 and 9 h were tested. Total and progressive motility, longevity and acrosomal integrity were measured and compared. The use of TriladylTM proved to result in better post-thaw parameters than the other two diluents. Equilibration times of between 4 and 9 h did not influence post-thaw sperm qualities significantly. For some of the treatments, exposure to semen extenders before freezing for less than 4 h resulted in inferior post-thaw semen parameters.

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 How to Increase Post-Thaw Semen Quality in Poor Freezing Stallions: Preliminary Results of the Promising Role of Seminal Plasma Added after Thawing

Animals ◽  
2019 ◽  
Vol 9 (7) ◽  
pp. 414 ◽  
Author(s):  
Jiří Šichtař ◽  
Filipa Bubeníčková ◽  
Jitka Sirohi ◽  
Ondřej Šimoník
Keyword(s):  
Plasma Membrane ◽  
Seminal Plasma ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Control Group ◽  
Computer Assisted ◽  
Kinematic Parameters ◽  
Prolonged Incubation ◽  
Sperm Analysis ◽  
Spermatozoa Motility

The aim of this study was to evaluate the effect of the addition of two types of seminal plasma (SP) after thawing on the functional characteristics of frozen–thawed (F–T) spermatozoa of poor freezing stallions during prolonged incubation periods. Seminal plasma from stallions with 35–40% (standard seminal plasma, (S-SP)) and 60–70% (above standard seminal plasma, (A-SP)) progressively motile spermatozoa after thawing was used. The motility, kinematic parameters (Computer Assisted Sperm Analysis), distribution of spermatozoa into subpopulations, integrity (carboxyfluorescein diacetate/propidium iodide staining), and functionality (hypo-osmotic swelling (HOS) test) of the spermatozoa plasma membrane were evaluated after thawing (T0) and after 30 min (T30) of incubation at 37 °C. There was no effect of SP addition on spermatozoa motility, but there was a significant positive effect on the kinematic parameters at T0 and T30. The addition of SP significantly increased the percentage of spermatozoa in the fast subpopulation at T0 as well as at T30. Plasma membrane integrity was not affected by the treatment, but functionality significantly decreased by 5% compared to the control group when samples were incubated for 30 min with A-SP. In conclusion, generally, the post-thaw addition of seminal plasma positively affected the post-thaw quality of semen from poor freezing stallions.

PSI-9 Evaluation of an Electronically-controlled Floor Cooling Pad on Semen Quality in Boars During Moderate Heat Stress

2021 ◽  
Vol 99 (Supplement_1) ◽  
pp. 222-223
Author(s):  
Hailey M Hedrick ◽  
Larissa K Shirley ◽  
Tyler Fields ◽  
Allan P Schinckel ◽  
Jay S Johnson ◽  
...  
Keyword(s):  
Heat Stress ◽  
Semen Quality ◽  
Sperm Concentration ◽  
Semen Parameters ◽  
Lower Percentage ◽  
Controlled Cooling ◽  
Study Objective ◽  
Progressive Motility ◽  
Motion Parameters ◽  
Kinematic Motion

Abstract Heat stress (HS) decreases semen quality and production in boars. Therefore, the study objective was to evaluate the use of electronically-controlled cooling pads to reduce the negative effects of HS on semen quality. Boars (n=24) were randomly allotted to two treatment groups: boars housed on a non-functional cooling pad (CON) or pads flushing water every 8 minutes or when the pad reached 28.5°C (FLUSH). For 3 d, boars were subjected to cyclical HS (28 to 32°C; &gt;50% relative humidity). Semen was collected for 7 weeks (2 weeks prior to determine baseline semen parameters, the day after HS, and weekly for 4 weeks post HS), and evaluated for volume, sperm concentration, motility, progressive motility, morphology, viability, and kinematic motion parameters. FLUSH boars had higher semen volumes compared to CON (P=0.011) without a corresponding increase in total sperm produced (P=0.852). Boars in FLUSH had higher motility in all weeks (87.0–90.8%) when compared to CON boars. FLUSH boars had higher motility starting in week 4 after HS compared to CON (P=0.017). No differences in progressive motility or kinematic motion parameters were found. There was a tendency for FLUSH boars to have a higher percent normal morphology compared to CON boars (83.1 vs 77.5%, P=0.083) resulting from decreased proximal and distal droplets in the FLUSH boars (P=0.029 and P=.0014, respectively). During week 2 post HS, there was a tendency for FLUSH boars to have a lower percentage of non-viable cells compared to all other weeks for FLUSH and CON boars (P=0.088). Cooling pads were effective at reducing the negative impacts of HS on semen volume, motility, and morphological abnormalities.

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