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.

scholarly journals 96 A NOVEL EXTENDER FOR PRESERVATION OF BACTRIAN CAMEL (CAMELUS BACTRIANUS) SEMEN

2005 ◽  
Vol 17 (2) ◽  
pp. 198 ◽  
Author(s):  
A. Niasari-Naslaji ◽  
S. Mosaferi ◽  
A.A. Gharahdaghi ◽  
A. Abarghani ◽  
A. Ghanbari ◽  
...  
Keyword(s):  
Semen Quality ◽  
Egg Yolk ◽  
Bactrian Camel ◽  
Dromedary Camel ◽  
Persian Language ◽  
Semen Collection ◽  
Arcsine Transformation ◽  
History Of ◽  
Collection Time ◽  
Better Than

Lactose and Green buffer (IMV, France) are the most commonly used extenders for camel semen. The viability of Bactrian camel spermatozoa in lactose extender is reduced after 4 h of incubation at 4°C (unpublished data). Although Green buffer is used for dromedary camel semen, there are no data indicating its effectiveness for Bactrian camel semen. More recently, we reported that the osmolality and pH of Bactrian camel semen are 316.1 ± 1.48 mOsm/kg and 7.4 ± 0.03, respectively. The objective of this study was to compare three different semen extenders, to determine if a TRIS-based diluent (SHOTOR* Diluent), a completely defined diluent, can maintain cooled camel sperm as effectively as established diluents. SHOTOR Diluent consists of 2.6 g TRIS, 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. SHOTOR Diluent, lactose, 10% (w/v), with an osmolality of 330 mOsm/kg and pH of 6.9, and Green buffer were compared in this study. All extenders contained 20% egg yolk. Semen was collected from bulls with a sound history of semen quality and fertility (n = 3), using a modified artificial vagina, and divided equally into the different extenders (Mosaferi S et al. 2004 15th Int. Cong. Anim. Reprod. 2, 520; Mosaferi S et al. 2004 Theriogenology, in press). Progressive forward motility and percentage of live spermatozoa were examined at the time of semen collection (time 0) and after 4, 12, and 24 h of incubation at 4°C. Data were analyzed using the GLM procedure in SAS/STAT after arcsine transformation. The forward progressive motilities of spermatozoa at 0, 4, 12, and 24 h after semen collection were 65.5, 54, 44.5, and 36.5% in SHOTOR Diluent; 31, 18.5, 8.5, and 0% in 10% lactose; and 60.5, 54.5, 33, and 32.5 % in Green buffer, respectively (Table 1). The percentage of live spermatozoa at 0, 4, 12, and 24 h were 84.5, 84, 81 and 74.5% in SHOTOR Diluent; 80, 79.5, 72.5, and 56.5% in 10% lactose; 89, 82.5, 82.5, and 77.5% in Green buffer, respectively (P > 0.05). The progressive forward motility of spermatozoa did not significantly decrease by 12 h at 4°C in SHOTOR Diluent (P > 0.05; Table 1), whereas it significantly decreased after 4 h and 12 h of incubation at 4°C in Green buffer and 10% lactose, respectively (P < 0.05; Table 1). Further decrease in the progressive forward motility occurred in all extenders after 24 h at 4°C (P < 0.05; Table 1). In conclusion, SHOTOR Diluent is better than Green buffer and 10% lactose as an extender for chilled storage of Bactrian camel semen for 12 hat4°C. Table 1. The progressive forward motility of Bactrian camel spermatozoa extended in SHOTOR Diluent (1), 10% lactose (2) and Green buffer (3) at the time of semen collection (time 0) and after 4, 12, and 24 h of incubation at 4°C *Shotor means camel in the Persian language. The authors wish to thank the director and station staff of Bactrian Camel Reseach Center at Meshkinshahr, Ardabil, for providing facilities and kind assistance throughout the experiment.

scholarly journals 94 COMPARING DIFFERENT LEVELS OF OSMOLARITY AND pH OF LACTOSE EXTENDER ON THE VIABILITY OF SPERMATOZOA IN THE BACTRIAN CAMEL (CAMELUS BACTRIANUS)

2005 ◽  
Vol 17 (2) ◽  
pp. 197 ◽  
Author(s):  
S. Mosaferi ◽  
A. Niasari-Naslaji ◽  
A.A. Gharahdaghi ◽  
A. Abarghani ◽  
A. Ghanbari ◽  
...  
Keyword(s):  
Semen Quality ◽  
Egg Yolk ◽  
Bactrian Camel ◽  
Mechanical Removal ◽  
Semen Collection ◽  
Arcsine Transformation ◽  
History Of ◽  
Semen Extender ◽  
Different Levels ◽  
Artificial Vagina

Lactose has been used widely as a semen extender for camels although in the absence of evidence illustrating its suitablility. Considering the osmolarity (316.1 ± 1.48 mOsm/kg) and pH (7.4 ± 0.03) of Bactrian camel semen (Mosaferi S et al. 2004 Theriogenology, in press), the objective of this study was to investigate the effect of osmolarity and pH of lactose extender on the viability of Bactrian camel spermatozoa. In Experiment I, with pH adjusted to 6.9, the effect of lactose concentrations of 9, 10, 11, 12, and 13% with an osmolarity of 290, 333, 350, 376, and 419 mOsm/kg, respectively, on the viability of spermatozoa was investigated. In Experiment II, with lactose fixed at 10%, the effect of extender with pH of 5.9, 6.9, 7.5, 7.9, and 8.9 on the viability of spermatozoa was examined. All extenders contained 20% egg yolk. In both experiments, semen was collected from camels with a sound history of semen quality and fertility (n = 3), using a modified artificial vagina, and divided into different treatments after mechanical removal of semen viscosity (Mosaferi et al. 2004). Progressive forward motility of spermatozoa was examined at the time of semen collection and at 4, 12, and 24 h after incubation at 4°C. Data were analyzed using the GLM procedure in SAS/STAT (SAS Institute, Inc., Cary, NC, USA) after arcsine transformation. At the time of semen dilution, the progressive forward motility of spermatozoa was greater at osmolarity of 290 (28.5%), 333 (34%), and 350 (31%) compared to 376 (13.5%) and 419 (1%) mOsm/kg (P < 0.05). The same trend in the progressive forward motility of spermatozoa was noticed after 4 h of incubation at 4°C; although a significant decrease (P < 0.05) occurred at 290 (11%), 333 (18 %) and 350 (16%). After 12 and 24 h of incubation at 4°C, the progressive forward motility of spermatozoa was less than 10% at 333 and 350 mOsm/kg (P < 0.05). At the time of semen dilution, the progressive forward motility of spermatozoa was greater (P < 0.05) at pH 6.9 (35.5%) and 7.5 (18%) compared to pH 5.9 (0%), 7.9 (7.5%) and 8.9 (2.5%). The same trend in the progressive forward motility of spermatozoa was observed after 4 h incubation at 4°C; although, a significant decrease (P < 0.05) occurred at pH 6.9 (15%) and 7.5 (9%) at this time. After 12 h incubation at 4°C, the progressive forward motility of spermatozoa was less than 5% at pH 6.9 and 7.5 (P < 0.05). In conclusion, 10% (333 mOsm/kg) and 11% (350 mOsm/kg) lactose, at the adjusted pH of 6.9, were the most suitable concentrations of lactose extender for preserving Bactrian camel semen for less than 4 h after which the viability of spermatozoa deteriorated significantly in this extender. The authors wish to thank the director and station staff of Bactrian Camel Research Center at Jahadabad, Meshkinshahr, Ardabil, for kind provision of facilities and assistance throughout the experiment.

scholarly journals Antioxidant Potential of Ferulic Acid on the Freezability of Bull Semen

2019 ◽  
Vol 4 (3) ◽  
pp. 1-4
Author(s):  
Omur AD
Keyword(s):  
Ferulic Acid ◽  
Liquid Nitrogen ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Single Step ◽  
Spermatozoa Motility ◽  
Bull Semen ◽  
Sperm Density ◽  
Significant Difference ◽  
Artificial Vagina

Ejaculates were collected twice a week from the bulls, via an artificial vagina, during two weeks. The suitable ejaculates obtained for sperm density (≥ 1.4 × 10 9 spermatoz oa / ml) and for motility (≥ 75%) were used for dilution and freezing of semen. A Tris - based extender (Tris 297.58mM, citric acid 96.32mM, fructose 82.66mM, egg yolk 15% (v/v), glycerol 5% (v/v), gentamicin 0.1 ml / 100ml, pH 6.8 - 7.0) was used as the base extender (cryopreservation diluent). Pooled ejaculate was split into 2 equal aliquots and diluted at 32 °C with base extender containing ferulic acid (100 μM) and no antioxidant (control), respectively. Each aliquot was diluted to a final semen concentrati on of approximately 1.2 × 10 8 sperm/ml (single step dilution), in 15 - ml polypropylene centrifuge tubes. After dilution, semen samples were kept at room temperature for 10 minutes then, the diluted semen samples were aspirated into 0.25 ml French straws, seal ed with polyvinyl alcohol powder and equilibrated at 5 °C for 3 h. After equilibration, the straws were frozen in liquid nitrogen vapour (4 cm above the liquid nitrogen, ~ - 100 o C ) for 10 min and then plunged into liquid nitrogen for storage, - 196 o C . In the study, sperm samples containing antioxidant and non - antioxidant were evaluated for spermatozoa motility and membrane integrity after freezing / thawing. In the present study, no statistically significant difference was found between the control and experim ental groups for motility and membrane integrity after freeze - thawing. The application consisted of 4 replications.

104 COMPARING DIFFERENT LEVELS OF OSMOLALITY OF SUCROSE EXTENDER ON THE VIABILITY OF SPERMATOZOA IN BACTRIAN CAMEL (CAMELUS BACTRIANUS)

2006 ◽  
Vol 18 (2) ◽  
pp. 160 ◽  
Author(s):  
S. Mosaferi ◽  
A. Niasari-Naslaji ◽  
N. Bahmani ◽  
A. A. Gharahdaghi ◽  
A. Abarghani ◽  
...  
Keyword(s):  
Semen Quality ◽  
Egg Yolk ◽  
Bactrian Camel ◽  
Dromedary Camel ◽  
Camelus Bactrianus ◽  
Semen Collection ◽  
History Of ◽  
Different Levels ◽  
Artificial Vagina

Disaccharides have been used as an extender for dromedary camel semen (Bravo et al. 2000 Anim. Reprod. Sci. 62, 173-193). More recently we have investigated the effect of different concentrations of lactose extender on the viability of Bactrian camel spermatozoa (Mosafer et al. 2005 Reprod. Fertil. Dev. 17, 197). Considering the osmolality (316.1 � 1.48 mOsm/kg) and pH (7.4 � 0.03) of Bactrian camel semen (Mosaferi et al. 2005 Theriogenology 63, 92-101), the objective of this study was to investigate the effect of osmolality of sucrose extender on the viability of Bactrian camel spermatozoa. Sucrose at the concentrations of 9, 10, 11, 12, and 13% with osmolalities of 292, 331, 356, 386, and 410 mOsm/kg, respectively, were prepared. All extenders contained 20% egg yolk and antibiotics, with pH adjusted to 6.9. Semen was collected from camels with a sound history of semen quality and fertility (n = 3) using a modified artificial vagina and divided into different treatments after mechanical reduction of semen viscosity (3). Progressive forward motility of spermatozoa was examined at the time of semen collection and at 4, 12, and 24 h after incubation at 4�C. Data were analyzed using the GLM procedure in SAS/STAT after arcsin transformation. At the time of semen dilution, the progressive forward motility of spermatozoa was greater at osmolality of 331 (23%) compared with 292 (1%), 386 (6%), and 410 (3.5%) mOsm/kg (P < 0.05). No progressive forward motility of spermatozoa was noticed after 4 h incubation at 4�C at osmolalities of 292, 386, and 410 mOsm/kg. At this time, a significant decrease (P < 0.05) of progressive forward motility occurred at osmolalities of 331 (4%) and 356 (0.5%) compared with that of the time of dilution. After 12 and 24 h incubation at 4�C, no progressive forward motility of spermatozoa was detected in any of these extenders. In conclusion, 10% sucrose (331 mOsm/kg) at the adjusted pH of 6.9 was the most suitable concentration of this disaccharide for preserving Bactrian camel semen for less than 4 h under chilled conditions.

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.

scholarly journals Effects of various diluents on the quality and shelf life of Donggala bull semen

2021 ◽  
Vol 902 (1) ◽  
pp. 012005
Author(s):  
Mirajuddin ◽  
Y Duma ◽  
M I Mumu ◽  
M R Ladjama ◽  
Nur A’fia ◽  
...  
Keyword(s):  
Citric Acid ◽  
Shelf Life ◽  
Storage Time ◽  
Egg Yolk ◽  
Sperm Viability ◽  
Decrease Rate ◽  
Bull Semen ◽  
And Storage ◽  
Artificial Vagina ◽  
Quality And Shelf Life

Abstract This study aimed to determine the effects of different semen diluent on the quality and storage time of liquid semen of Donggala bull. Semen was obtained from four selected bulls which collected using artificial vagina. The semen diluent is based on Tris aminomethane-citric acid with different concentration of glucose namely P1=0.00g, P2=0.25g, P3=0.50g, and P4=0.75g. Another group of treatment also prepared based on Tris aminomethane-glucose with different concentration of egg yolk namely P5=0%, P6=15%, P7=20%, and P8=25%. The data obtained were then analyzed descriptively. Results showed tris aminomethane-citric acid-glucose diluents had sperm progressive motile at >40% until the day 4 of storage, and tris aminomethane-glucose-egg yolk only able to support sperm life for 2 days, and in P8 group shows 41.02% motile on the day 3. In this study, we found the pattern of the increasing of shelf life affect to the decrease rate of sperm viability and normality.

scholarly journals Ram semen preserved at 0°C with soybean lecithin Tris-based extender substituted for egg yolk

2021 ◽  
Vol 34 (2) ◽  
pp. 192-197
Author(s):  
Jian-qing Zhao ◽  
Guo-liang Xiao ◽  
Wen-liang Zhu ◽  
Di Fang ◽  
Na Li ◽  
...  
Keyword(s):  
Soybean Lecithin ◽  
Egg Yolk ◽  
Control Group ◽  
Progressive Motility ◽  
Significant Difference ◽  
Acrosome Integrity ◽  
Normal Offspring ◽  
And Control ◽  
Ram Sperm ◽  
Artificial Vagina

Objective: The present study evaluated the preservation of ram semen at 0°C using soybean lecithin with a Tris-fructose extender.Methods: Semen was collected by artificial vagina ejaculation from six rams with proven fertility. High quality ejaculates were diluted by soybean lecithin (0.25%, 0.5%, 0.75%, 1.0%, 1.25%) using Tris-fructose extender and control (Tris-fructose egg yolk extender), respectively. The ejaculates were diluted to a concentration of 5×10<sup>8</sup> sperm/mL, followed by cooling to 0°C in 90 min and maintaining the temperature for 12 days. The diluted semen samples were examined and recorded for sperm progressive motility, acrosome integrity at 0, 24, 72, 144, 216, 288 h, respectively. Two hundred and twenty-three ewes were inseminated for 216 h with optimal soybean lecithin concentrated semen or control via trans-cervical insemination.Results: The results showed that there were no differences in sperm progressive motility at 0, 24, 72, and 144 h (p>0.05). After 216 h, the sperm progressive motility in the control group and 0.5% concentration groups was significantly higher when compared to 0.25% concentration (p<0.05). The 0.5% concentration group demonstrated the highest survival rate and had no difference with the control group (p>0.05). At 216 h, the sperm progressive motility of all groups was still above 50%. The acrosome integrity of all groups was decreased with prolongation of storage time, but there was no difference at each time point (p>0.05). There was no significant difference in the lambing rate and pregnancy rate between the 0.5% concentration group and the control group (p>0.05).Conclusion: These results suggest that ram sperm is capable of fertilization after preservation at 0°C with 0.5% of soybean lecithin in Tris-based extender substituted for egg yolk and produce normal offspring after insemination.

25 A preliminary study of the effects of breed and nanowater as extender diluent on ram semen characteristics post-thawing

2020 ◽  
Vol 32 (2) ◽  
pp. 138
Author(s):  
M. Murawski ◽  
J. Szymanowicz ◽  
T. Schwarz ◽  
P. Bartlewski
Keyword(s):  
Sperm Morphology ◽  
Egg Yolk ◽  
Plasma Generator ◽  
Deionized Water ◽  
Ex Situ ◽  
Survival Times ◽  
Becton Dickinson ◽  
Significant Difference ◽  
Semen Extender ◽  
Difference Test

This study examined the effect of genotype and extender diluent on the characteristics of frozen-thawed ram semen. Twenty ejaculates collected from 10 rams aged 4-12 years (2 Polish Lowland (PON), 4 synthetic line BCP (Berrichon du Cher×Charolais×PON/Polish Merino), and 4 SCP (Suffolk×Charolais×PON/Polish Merino) in mid-breeding season were divided into six equal portions each and frozen in semen extender prepared by mixing commercial Triladyl extender (MiniTub GmbH) with deionized water (Aqua Purificata, Prolab) or nanowater (NW; deionized water declusterized in a cold plasma generator for 15, 30, 45, 60, or 90 minutes (15’, 30’, 45’, 60’, or 90’, respectively); Nanotechnology Systems) and Gallus domesticus egg yolk (1:3:1 vol/vol/vol). All semen samples were evaluated for progressive motility (Sperm Class Analyzer) and sperm morphology (Nikon Eclipse 80i microscope, Nikon Corp.). In addition, the ex situ survival time at 37°C and extender content of ALT (alanine transferase), ALP (alkaline phosphatase), and aspartate aminotransferase were measured, and the numbers of apoptotic, necrotic, and live spermatozoa were determined post-thawing with flow cytometry (BD Accuri™ C6 Plus, Becton Dickinson). Data were analysed by two-way analysis of variance and least significant difference test (SigmaPlot, Systat Software Inc.). The PON exceeded (P&lt;0.05) SCP rams in the occurrence of mid-piece and double tail defects using NW30’ and NW45’ extenders. The proportion of spermatozoa with proximal droplets was greater (P&lt;0.05) in PON than in SCP or BCP rams with NW15’ and NW30’ extenders, respectively, but the percentage of sperm with detached heads was greater (P&lt;0.05) in BCP and SCP compared with PON using NW15’ extender. Overall, spermatozoa in PON rams’ semen had survival times greater by 25-50min (P&lt;0.05) compared with those for semen from BCP and SCP synthetic lines. The ALT levels were significantly elevated in SCP compared with PON and BCP rams only in NW90’ extender. The numbers of necrotic spermatozoa were greater (P&lt;0.05) in SCP and BCP compared with PON rams, and the numbers of apoptotic spermatozoa were greater (P&lt;0.05) in BCP than in SCP rams with NW45’, NW60’, and NW90’ extenders. Live spermatozoa were more prevalent (P&lt;0.05) in PON than in BCP rams in NW45’ extender post-thawing. To summarise, semen from PON rams had higher survival times and fewer spermatozoa with detached heads but more mid-piece defects and sperm with double tails post-thawing compared with BCP and SCP breeds. The number of necrotic cells was greater in semen from BCP and SCP rams compared with PON. It can be concluded that breed-related differences in post-thaw sperm morphology were apparent in extenders containing NW15-60’, whereas the variations in cell necrosis/apoptosis were primarily seen with the use of NW45-90’ extenders.

scholarly journals Kualitas Sperma Beku Sapi Bali dalam Pengencer Air Kelapa Modifikasi dengan Berbagai Aras Dimethyl Sulfoxide (FROZEN SPERM QUALITY OF BALI BULLS IN MODIFIED COCONUT WATER EXTENDER WITH DIFFERENT DIMETHYL SULFOXIDE CONCENTRATION)

2019 ◽  
Vol 20 (1) ◽  
pp. 93
Author(s):  
Thomas Mata Hine ◽  
Kirenius Uly ◽  
Wilmientje Marlene Nalley ◽  
Heri Armadianto
Keyword(s):  
Dimethyl Sulfoxide ◽  
Liquid Nitrogen ◽  
Sperm Quality ◽  
Egg Yolk ◽  
Optimal Concentration ◽  
Coconut Water ◽  
Low Molecular Weight ◽  
Frozen Semen ◽  
Artificial Vagina

Dimethyl Sulfoxide (DMSO) is one type of cryoprotectant which has a low molecular weight so that it is easier to enter cells when cryopreservation. The purpose of this study was to explore the optimal concentration of DMSO in modified coconut water (mCW) extender that were able to maintain frozen sperm quality of bali bulls. Semen was collected from two four-year old bali bulls by artificial vagina. Good quality semen diluted with mCW (young coconut water + 20% egg yolk + 7.5 % moringa leaf extract) and supplemented by 3, 5, or 7% DMSO. Semen was filled into 0.25 ml ministraw, and was incubated in a refrigerator at 5°C for four hours, frozen on the surface of liquid nitrogen for 10 minutes and then dipped into liquid nitrogen. The quality of post thawing sperm was measured 24 hours later by placing the ministraw of frozen semen into water at 37oC for 30 seconds. Data were analyzed by analysis of variance and continued with Duncan test. Postthawing observations showed that bali bulls sperm cryopreserved at 3% DMSO yielded higher motility and viability (p<0.05) i.e. 36 and 44.15%, than DMSO 5% i.e. 18 and 23.65%, and DMSO 7% i.e. 7 and 12.62%. The recovery rate of sperm cryopreserved at 3% DMSO was also higher (p<0.05) than DMSO 5 and 7%, successively 45.65, 23.06, and 8.86%. The results of this study concluded that the optimal concentration of DMSO in mCW diluent to maintain frozen sperm quality of bali bulls was 3%. 

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.

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