scholarly journals PENAMBAHAN L – ARGININ DALAM PENGENCER TRIS KUNING TELUR SETELAH EKUILIBRASI 2 JAM TERHADAP MOTILITAS DAN MEMBRAN PLASMA UTUH SPERMATOZOA DOMBA SAPUDI

2020 ◽  
Vol 7 (2) ◽  
pp. 86
Author(s):  
Hastini Suryaningsih ◽  
Emy Koestanty Sabdoningrum ◽  
Suherni Susilowati ◽  
Trilas Sardjito ◽  
Wurlina Wurlina ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Analysis Of Variance ◽  
Sperm Motility ◽  
Egg Yolk ◽  
Equilibration Time ◽  
Sperm Plasma Membrane ◽  
Artificial Vagina ◽  
Plasm Membrane

This research was aimed to determine the addition of L-arginine on motility and sperm plasma membrane of Sapudi sheep after two hour equilibration time in egg yolk tris diluent. This research used fresh samples of sheep Sapudi semen collected by artificial vagina, then were devided into 4 treatments and 6 replications. Analysis of the data using Analysis of Variance (ANOVA) then proceed to the Duncan Test to determine significant differences between treatments. The first treatment P0 was no L – Arginine added as control. P1 was treated with 0,004 M L – Arginine. P2 was treated with 0,005 M L – Arginine, P3 was treated with 0,006 M L – Arginine. Result of two hour equilibration time for the sperm motility showed: (P0) 46,67a   5,16, (P1) 49,16ab   5,84, (P2) 55,00bc   7,74, (P3) 58,33c   6,83. Result of two hour equilibration time for the sperm plasm membrane were: (P0) 39,33a   4,92, (P1) 43,33ab   4,22, (P2) 46,50 bc   3,44, (P3) 50,83 c   3,48. The addition of L-arginine with a concentration as much as 0.006 M, shows the highest result in sperm motility and sperm plasm membrane intact of Sapudi semen.

187 EXPLOITATION OF IN VITRO CAPACITATION FOR NANOPARTICLE INCORPORATION WITHIN MAMMALIAN SPERMATOZOA

2016 ◽  
Vol 28 (2) ◽  
pp. 224
Author(s):  
L. Myles ◽  
C. Durfey ◽  
P. Ryan ◽  
S. Willard ◽  
J. Feugang
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Surface Membrane ◽  
Milk Powder ◽  
Noninvasive Evaluation ◽  
Control Group ◽  
Body Tissues ◽  
Sperm Plasma Membrane

Migration and interactions of mammalian gametes occur in deep body tissues after mating, rendering difficult any in situ noninvasive evaluation of their performances with current methods. In our effort to develop an effective and real-time in vivo imaging approach, we have successfully labelled porcine gametes with self-illuminating bioluminescent and red-shifted quantum dot nanoparticles (QD) in our previous studies (Feugang et al. 2012 J. Nanobiotechnol. 10, 45; Feugang et al. 2015, J. Nanobiotechnol. 13, 38). The present effort aimed at investigating whether QD could be incorporated into spermatozoa through induced in vitro capacitation, which increases sperm plasma membrane fluidity. Fresh extended boar semen was placed on top of a Percoll gradient and centrifuged. Purified motile spermatozoa were collected and washed with pre-warmed PBS. Pelleted spermatozoa were resuspended in the modified Tris-buffered medium with BSA fraction-V (1 mg mL–1; modified Tween medium B with milk powder and BSA). Sperm aliquots (108) were supplemented or not (control) with QD only (QD+; 1 nM), QD+caffeine (2 mM), or QD+heparin (10 µg mL–1); with caffeine and heparin being used as routine capacitant agents in fertilization media. All aliquots were incubated at 38.5°C, under 5% CO2 for 0.5, 1, or 3 h. Spermatozoa were then analysed for motility characteristics and imaged for confirmation of QD-sperm interactions (bioluminescence emission) and localization (transmission electron microscope; TEM). Motility data of 5 replicates were analysed with ANOVA-2, and P < 0.05 was set as threshold of significance. Total sperm motility (TSM) significantly improved with the presence of either or both QDs and capacitant agents after 0.5 and 1 h incubations. With exception of the QD+heparin, all other groups had significantly decreased TSM after 3 h of incubation, when compared with TSM at 0.5 and 1 h. Higher proportions of progressive and rapid (≥45 µm s–1) spermatozoa were observed in the presence of both capacitant agents (P < 0.05), and only QD+heparin maintained greater proportions after 3 h. Sperm straight-line velocity significantly increased in the QD+caffeine at 0.5 h and in both QD+caffeine and QD+heparin thereafter. Sperm straightness data were increased by both caffeine and heparin during incubations. Strong bioluminescence signals were observed in spermatozoa incubated with QDs compared to the background signal seen in the control group. The TEM images revealed consistent surface membrane attachment of QDs in all QD+ groups, whereas transmembrane and intra-spermatic localizations were visible in both QD+caffeine and QD+heparin groups. We concluded that supplementations of medium containing QDs with caffeine or heparin allow the crossing of sperm plasma membrane by QD. No toxic effect of QD on sperm motility was observed, which confirmed our previous report using a similar ratio of QDs over spermatozoa. Exploration of efficient incorporation of QD into spermatozoa as a promising approach for noninvasive molecular imaging is still ongoing, as well as further sperm viability assessments. Supported by the NIH grant #5T35OD010432 and USDA-ARS Biophotonics Initiative grant #58–6402–3-0120.

scholarly journals MIG-23 is involved in sperm migration in Ascaris suum

2021 ◽  
Author(s):  
Xia Wang ◽  
Qiushi Wang ◽  
Ruijun He ◽  
Qi Zhang ◽  
Jin Shan ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Atpase Activity ◽  
Sperm Motility ◽  
Ascaris Suum ◽  
Nucleoside Triphosphate ◽  
Mitochondrial Activity ◽  
Major Sperm Protein ◽  
Sperm Activation ◽  
Sperm Migration ◽  
Sperm Plasma Membrane

Sperm motility acquisition during maturation is essential for successful fertilization.Extracellular adenosine-5'-triphosphate (ATP) level mediation by MIG-23, which is a homolog of human ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase), was required for major sperm protein filament dynamics and sperm motility in the nematode Ascaris suum. MIG-23 was localized on the sperm plasma membrane. During sperm activation, mitochondrial activity was increased dramatically, and a large amount of ATP was produced and stored in refringent granules (RGs). In addition, a portion of the produced ATP was released to the extracellular space through ATP channels, which were composed of innexins and localized on the sperm plasma membrane. Spermatozoa, instead of spermatids, hydrolyzed exogenous ATP and processed ecto-ATPase activity. MIG-23 contributed to the ecto-ATPase activity of spermatozoa. MIG-23 activity was interrupted, spermatozoa also decreased their ATP hydrolysis activity. Blocking MIG-23 activity resulted in an increase in the depolymerization rate of MSP filaments in pseudopodia, which eventually affected nematode sperm migration. Overall, our data imply that MIG-23, which contributes to the ecto-ATPase activity of spermatozoa, regulates sperm migration by modulating extracellular ATP levels.

scholarly journals Effects of the addition of docosahexaenoic acid and ?-tocopherol on quality of equine spermatozoa stored at 5°C

2020 ◽  
Vol 41 (1) ◽  
pp. 167 ◽  
Author(s):  
Breno Fernandes Barreto Sampaio ◽  
Bruno Gomes Nogueira ◽  
Maria Inês Lenz Souza ◽  
Eliane Vianna da Costa-e-Silva ◽  
Carmem Estefânia Serra Neto Zúccari
Keyword(s):  
Lipid Peroxidation ◽  
Plasma Membrane ◽  
Docosahexaenoic Acid ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Membrane Composition ◽  
Artificial Vagina

Plasma membrane composition has impact on phase transition from liquid crystal to gel state of cooled sperm cell. The incorporation of polyunsaturated fatty acids increases its fluidity and can contribute to sperm motility. The aim of this study was to compare the effect of adding docosahexaenoic acid (DHA) and ?-tocopherol (?-Toh) to the cooling extender, singly or combined, to the equine sperm parameters, submitted to cooling, up to 72 hours. Two ejaculates of ten stallions collected with artificial vagina were used, and evaluated for motility, plasma membrane integrity, chromatin fragmentation, mitochondrial activity and lipid peroxidation, according to the following treatments: C; DHA; ?-Toh; DHA/?-Toh; EtOH 100: and EtOH 140 (corresponding to control; 10 ng mL-1 of DHA; 2 mM of ?-Toh; : 10 ng mL-1 of DHA + 2 mM of ?-Toh; 100 µL of ethanol and 140 µL of ethanol respectively). DHA treatment showed higher motility (68.2 ± 12.3; p < 0.05) when compared to control (62.1 ± 16.2), DHA/?-Toh (61.3 ± 12.7) and EtOH (58.1 ± 8.6) groups. In lipid peroxidation assay, the control group showed 2,506.2 ± 796.4 ng of MDA 108 spermatozoa-1, being significantly higher (p < 0.05) than the groups treated with DHA (2,036.0 ± 687.0), ?-Toh (1,890.8 ± 749.5) and DHA/?-Toh (1,821.1 ± 627.2). In conclusion, ?-Toh was effective in diminishing lipid peroxidation of equine sperm subjected to cooling, and DHA improved sperm motility and, in spite of being a polyunsaturated fatty acid with high susceptibility to peroxidation, reduced lipid peroxidation.

scholarly journals Peningkatan Motilitas Spermatozoa Kambing Nubian Setelah Pemberian PGF2α dalam Pengencer Andromed

2020 ◽  
Vol 20 (1) ◽  
pp. 32-37
Author(s):  
Adhea Prestiya ◽  
Tongku Nizwan Siregar ◽  
Husnurrizal Husnurrizal ◽  
Sri Wahyuni ◽  
Eka Meutia Sari ◽  
...  
Keyword(s):  
Analysis Of Variance ◽  
Sperm Motility ◽  
Statistical Tests ◽  
Spermatozoa Motility ◽  
Artificial Vagina

ABSTRAK. Penelitian ini bertujuan mengetahui pengaruh pemberian PGF2α dalam pengencer semen komersial (Andromed) terhadap peningkatan motilitas spermatozoa kambing Nubian. Penelitian ini menggunakan sampel semen yang dikoleksi dari 3 ekor kambing Nubian berumur 2-3 tahun menggunakan vagina buatan dan dievaluasi kualitasnya secara makroskopis dan mikroskopis. Setelah dievaluasi, sampel semen ditambahkan pengencer Andromed lalu dibagi atas 3 kelompok perlakuan, yaitu: P1; P2; dan P3 yang masing-masing ditambahkan NaCl fisiologis; 37,5 µg PGF2α; dan 75 µg PGF2α. Seluruh sampel disimpan dalam refrigerator selama 4 jam dan dilakukan pemeriksaan motilitas spermatozoa. Data yang diperoleh dianalisis dengan sidik ragam pola satu arah (ANOVA) dan dilanjutkan dengan uji Duncan. Hasil penelitian menunjukkan, bahwa motilitas spermatozoa (%) kambing Nubian pada P1; P2; dan P3 masing-masing adalah 26,33±5,5; 62,0±3,5; dan61,8±10,13 (P0,05). Disimpulkan bahwa penambahan PGF2α pada pengencer Andromed dapat meningkatkan motilitas spermatozoa kambing Nubian.  (The improvement of sperm motility in Nubian goat after PGF2α administration in andromed semen diluents) ABSTRACT. The study aims to determine the administration effect of PGF2α in a commercial semen diluents (Andromed) on improvement of Nubian goat sperm motility. This study used semen samples that collected from three Nubian goats aged 2-3 using artificial vagina and their quality evaluated macroscopically and microscopically. After evaluated, semen samples were added with Andromed diluents then divided into three groups (P1, P2, and P3) where each group was then added with 0,9% physiologic NaCl, 37.5 µg PGF2α, and 75 µg PGF2α, respectively and stored in a refrigerator for 4 hours and subsequently spermatozoa motility was examined. The data obtained were analyzed by one-way analysis of variance (ANOVA) and followed by Duncan test. The results showed that the spermatozoa motility (%) of Nubian goats at P1, P2, and P3 were 26.33±5.5, 62.0±3.5, and 61.8±10.13, respectively. Based on the statistical tests showed that the administration of PGF2α at P2 and P3 had a significant effect (P0,05) on the motility of spermatozoa of Nubian goats, but the motility decreased in P1. The conclusion of this study is the addition of PGF2α to Andromed diluents can increase the motility of spermatozoa of Nubian goats.

scholarly journals Efficiency of Commercial Egg Yolk-Free and Egg Yolk-Supplemented Tris-Based Extenders for Dromedary Camel Semen Cryopreservation

Animals ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 999 ◽  
Author(s):  
Ayman Abdel-Aziz Swelum ◽  
Islam M. Saadeldin ◽  
Hani Ba-Awadh ◽  
Mohsen G. Al-Mutary ◽  
Abdullah F. Moumen ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Dna Integrity ◽  
Computer Assisted ◽  
Dromedary Camel ◽  
Plasma Membrane Integrity ◽  
Semen Cryopreservation

This study compared the efficiency of commercial egg yolk-free (AndroMed, OPTIXcell) and egg yolk-supplemented (Triladyl, Steridyl) Tris-based extenders for semen cryopreservation in seven adult dromedary camels. The camel-specific extender SHOTOR was used as control. The collected semen samples were evaluated and diluted with SHOTOR, Triladyl, Steridyl, AndroMed, or OPTIXcell. The diluted semen was gradually cooled and equilibrated for two hours before liquid nitrogen freezing. Semen was evaluated prior to freezing and after freeze-thawing cycles for motility, kinetics, vitality, abnormality, plasma membrane integrity, and DNA fragmentation using computer-assisted sperm analysis. In pre-freezing evaluation, progressive sperm motility was higher in SHOTOR-diluted samples (21.54 ± 1.83) than in samples diluted with Steridyl, OPTIXcell, or AndroMed (15.76 ± 1.80, 17.43 ± 1.10, and 13.27 ± 1.07, respectively). Moreover, Triladyl and SHOTOR resulted in significantly (p < 0.05) better sperm vitality and DNA integrity than all other diluents, but Triladyl resulted in a significantly (p < 0.05) better plasma membrane integrity (87.77 ± 0.31) than SHOTOR (85.48 ± 0.58). In the post-thawing evaluation, Triladyl led to significantly (p < 0.05) higher sperm motility (38.63 ± 0.81%; p < 0.05) when compared to SHOTOR, Steridyl or AndroMed (35.09 ± 1.341%, 34.4 ± 0.84%, and 31.99 ± 1.48%, respectively), with OPTIXcell being the least efficient (28.39 ± 0.86%). Progressive sperm motility was the highest when using Triladyl. Post-thawing curvilinear, straight line and average path sperm velocities were highest with Triladyl and lowest with AndroMed. Triladyl led to the highest linearity coefficient and straightness sperm coefficient, while SHOTOR to the highest DNA and plasma membrane integrity. OPTIXcell and AndroMed resulted in poor post-thawing sperm vitality, while Steridyl was less efficient than Triladyl. The highest rate of sperm abnormalities was recorded with OPTIXcell and the lowest with SHOTOR or Triladyl. In conclusion, SHOTOR, Triladyl, Steridyl, AndroMed, and OPTIXcell can all be used for camel semen cryopreservation; however, SHOTOR and Triladyl provided the best post-thawing sperm quality. Based on our findings, Triladyl is the best commercially available extender for dromedary camel semen cryopreservation to date.

105 COMPARISON BETWEEN TWO EXTENDERS FOR CRYOPRESERVATION OF BACTRIAN CAMEL SEMEN

2006 ◽  
Vol 18 (2) ◽  
pp. 161
Author(s):  
A. Niasari-Naslaji ◽  
S. Mosaferi ◽  
A. A. Gharahdaghi ◽  
A. Abarghani ◽  
A. Ghanbari ◽  
...  
Keyword(s):  
Citric Acid ◽  
Liquid Nitrogen ◽  
Egg Yolk ◽  
Deionized Water ◽  
Bactrian Camel ◽  
Equilibration Time ◽  
Persian Language ◽  
Significant Difference ◽  
Arcsine Transformation ◽  
Artificial Vagina

A Tris-based extender (SHOTOR diluent) has been developed for preserving Bactrian camel semen at 4�C (Niasari-Naslaji et al. 2005 Reprod. Fertil. Dev. 17, 198 (abstr.)). The present study investigated the possibility of utilizing the SHOTOR diluent for the cryopreservation of Bactrian camel semen. A modified bovine artificial vagina (Masaferi et al. 2005 Theriogeology 63, 92-101) was used to collect semen from three fertile bulls. The viscosity of the semen was reduced mechanically (Mosateri et al. 2005) and the homogenized semen was divided equally into two parts. Each part was sequentially diluted with either IMV buffers (Green buffer: first extender; White buffer: second extender; IMV, France) or SHOTOR diluents (without glycerol: first extender; with 12% glycerol: second extender). SHOTOR diluent consists of 2.6 g TIS, 1.35 g citric acid, 1.2 g glucose, and 0.9 g fructose in 100 mL of deionized water, with an osmolality of 330 mOsm/kg and pH of 6.9. All extenders had 20% egg yolk and antibiotics. The semen was diluted at the ratio of 1:1 with the first extender. The diluted semen was then cooled within 2 h to 4�C. At this temperature, the second extender was added at the same volume as the diluted semen in three steps with an equal volume, 10 min apart. After a 30-min equilibration time, beginning after addition of the last fraction of the second extender, the diluted semen was loaded into 0.5-mL straws at a concentration of 50 � 106 sperm per straw. The straws were maintained for 20 min at 4 cm above the liquid nitrogen surface, after which they were plunged into liquid nitrogen. The semen was thawed at 40�C water bath for 20 s. Progressive forward motility of spermatozoa was assessed at the time of dilution and immediately after thawing of the semen. The experiment was replicated four times. Data were analyzed using GLM procedure in SAS/STAT after arcsine transformation. At the time of dilution, there was no significant difference in progressive forward motility of spermatozoa between IMV buffers (51.8%) and SHOTOR diluent (61%; P > 0.05). However, after thawing, there was a significant decrease in progressive forward motility of spermatozoa in IMV buffers (4.2%) compared to SHOTOR diluent (29.9%, P < 0.05). In conclusion, in this experiment, SHOTOR diluent was more efficient for cryopreserving Bactrian camel semen than IMV extender. Shotor means camel in the Persian language.

21 Effect of egg yolk extracted low-density lipoprotein on cryopreserved Nguni bull semen

2019 ◽  
Vol 31 (1) ◽  
pp. 136
Author(s):  
M. M. Tshabalala ◽  
K. A. Nephawe ◽  
M. L. Mphaphathi ◽  
C. M. Pilane ◽  
T. L. Nedambale
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Egg Yolk ◽  
Semen Parameters ◽  
Dna Integrity ◽  
Low Density ◽  
Bull Semen ◽  
Frozen Semen ◽  
Live Sperm

Egg yolk has been reported to have a beneficial effect on sperm quality following cryopreservation, and this led to its widespread use in semen extenders. However, egg yolk contains substances that inhibit respiration of sperm cells and diminish their motility rate. Moreover, it also contains low-density lipoproteins (LDL) that have a protective effect on sperm during the cryopreservation process. The objective of this study was improve cryopreservation of Nguni bull semen using egg yolk low-density protein. A total of 25 ejaculates were collected from 5 Nguni bulls aged 4 to 5 years using an electroejaculator during the natural breeding season. Collected raw semen samples were transported to the laboratory and evaluated for sperm motility, viability, plasma membrane, acrosome, and DNA integrity before dilution. Semen was randomly diluted with a sodium citrate extender supplemented with 20% egg yolk (control) and with 6, 8, 10, and 12% LDL concentrations. The diluted semen sample groups were equilibrated for 4h at 5°C. Following equilibration, semen was loaded into 0.25-mL straws and frozen in a controlled-rate programmable freezer. The groups of semen straws were then plunged into LN and transferred into LN tanks (−196°C) for storage. The frozen semen straws per treatment group were thawed at 37°C and evaluated for sperm motility, viability, plasma membrane, acrosome, and DNA integrity. Data were analysed with ANOVA using Stata V12 statistical software (StataCorp., College Station, TX), and treatment means were separated using Fisher’s protected t-test at the significant level of P&lt;0.05. Sperm motility and membrane integrity were significantly higher (P&lt;0.05) in frozen-thawed semen diluted with 8% LDL compared with the other concentrations. However, 6 and 8% LDL resulted in a significantly higher (P&lt;0.05) live sperm, DNA, and acrosome integrity. Frozen-thawed semen diluted with 10 and 12% LDL resulted in the lowest percentages of sperm motility, live sperm, plasma membrane, acrosome, and DNA integrity following cryopreservation. In conclusion, extender containing 8% LDL resulted in improved Nguni bull semen parameters such as sperm motility, viability, plasma membrane, acrosome, and DNA integrity following cryopreservation. Further studies are required to determine the fertilizing capacity of semen diluted and frozen with LDL in vitro and in vivo.

Effect of different levels of egg-yolk on freezability of Jakhrana buck semen

2016 ◽  
Author(s):  
Narendra Kumar ◽  
B. Rai ◽  
Chetna Gangwar ◽  
S. A. Lone ◽  
Anshuman Kumar ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Egg Yolk ◽  
Osmotic Swelling ◽  
Semen Collection ◽  
Sperm Abnormality ◽  
Before And After ◽  
Different Levels ◽  
Artificial Vagina ◽  
Intensive System ◽  
Acrosomal Integrity

The present study was designed to determine the effect of different levels of egg-yolk on freezability of Jakhrana buck semen. Six healthy Jakhrana bucks (BW=30 ± 2kg, age=12 ± 0.5 month) were used for semen collection. These bucks were maintained under semi-intensive system at Jakhrana Unit of C.I.R.G. Makhdoom, Mathura. A total of 48 ejaculates (6 bucks × 8 replicates) were collected twice a week using artificial vagina. Each ejaculate was divided into 4 groups (G1, G2, G3 and G4). The G1, G2, G3 and G4 were extended with Tris-egg yolk-citric acid- fructose-glycerol (TEYCFG) extenders containing 5, 10, 15 and 20% egg yolk level, respectively. Each ejaculate was evaluated for sperm motility, viability, abnormality, and hypo-osmotic swelling (HOS) response and acrosome integrity before and after freezing. At pre-freeze stage no significant (P>0.05) difference in sperm motility and viability was found among all groups. Sperm abnormality was significantly (P<0.05) higher in G4 as compared to other groups (G1, G2, G3). The HOS response and acrosomal integrity was significantly (P<0.05) higher in G1, G2 and G3 as compared to G4. However, no significant (P>0.05) difference was observed in HOS response and acrosomal integrity among G1, G2 and G3. At post thaw stage, sperm motility, viability and HOS response was significantly (P<0.05) higher in G1 and G2 as compared to G3 and G4. Sperm abnormality was significantly (P<0.05) lower in G2 as compared to other groups. The acrosomal integrity was significantly (P<0.01) higher in G1 and G2 as compared to G3 and G4. It is concluded that 10% egg yolk in Tris based extender may be the best for successful cryopreservation of Jakhrana buck semen.

Plasma membrane changes during the liquid storage of boar spermatozoa: A comparison of methods

2010 ◽  
Vol 58 (1) ◽  
pp. 105-116 ◽  
Author(s):  
Dariusz Gączarzewicz ◽  
Małgorzata Piasecka ◽  
Jan Udała ◽  
Barbara Błaszczyk ◽  
Tomasz Stankiewicz ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Semen Analysis ◽  
Membrane Integrity ◽  
Live Cells ◽  
Lower Percentage ◽  
Loss Of Function ◽  
Plasma Membrane Integrity ◽  
Sperm Plasma Membrane ◽  
Semen Preservation

Studies were performed on boar semen routinely used at the local artificial insemination (AI) centre. The semen was stored in a Safe Cell Plus commercial extender at 17 °C for nine days. The aim of our research was focused on changes in sperm plasma membrane integrity. The integrity of the sperm plasma membrane and acrosome as well as sperm motility decreased after dilution and during storage of the semen. The highest percentage of live sperm was identified by the eosin-nigrosin method, a lower percentage by the SYBR-14/PI test, and the lowest percentage of live cells was discovered by the hypoosmotic swelling (HOS) test (P < 0.01). There were significant differences between the results of staining methods and sperm motility (P < 0.01). No significant differences were found between the HOS test results and sperm motility. The plasma membrane integrity parameters positively correlated (P < 0.001) with each other and with sperm motility but negatively with aspartate aminotransferase activity. Our findings confirmed that the boar sperm aging changes, which increased during liquid semen preservation, were connected with the loss of function and integrity of the sperm plasma membrane. The employed complementary tests are comprehensive indicators of sperm membrane integrity during long-term semen preservation, and they can help establish the actual number of ‘healthy’ cells. The assays may be used in AI laboratories and should be incorporated into the routine of semen analysis.

scholarly journals FREEZING CAPABILITY OF PASUNDAN BULL SPERM USING TRIS-EGG YOLK, TRIS-SOY, AND ANDROMED® DILUENTS

2017 ◽  
Vol 11 (1) ◽  
pp. 45-49
Author(s):  
Abdullah Baharum ◽  
R. Iis Arifiantini ◽  
Tuty Laswardi Yusuf
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Sperm Concentration ◽  
Egg Yolk ◽  
Freezing Process ◽  
Sperm Abnormalities ◽  
Bull Sperm ◽  
Motility Of Sperm ◽  
Artificial Vagina

The aims of this study were to investigate the freezing capability of Pasundan bull spermatozoa in Tris-egg yolk (TEY), Tris-soy (TS), and AndroMed® as diluents. Semen were collected twice a week from four Pasundan bulls aged 3-5 years old using an artificial vagina and evaluated macro- and microscopically. Semen had ≥70% sperm motility, ≥800x106/mL sperm concentration, and less than 20% sperm abnormalities were divided into three parts and each of them diluted with TEY, TS, or AndroMed®. After an equilibration step at 5°C for four hours, diluted semen were packaged in 0.25 mL straw, frozen in liquid nitrogen for ten minutes and kept in liquid nitrogen container until examination. Motility test on fresh, diluted, equilibrated, and after-thawed semen was done using Androvision®. The results showed that after thawing motility of sperm diluted in AndroMed® (58.64±0.72%) was higher than in TEY (49.45±1.22%) and TS (39.34±6.33%). Sperm motility of Pasundan bulls diluted in these three diluents reduced around 33.27±2.45% during freezing process.

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