Effect of Dimethyl sulfoxide (DMSO) and egg yolk on sperm motility, fertility and hatching rates of depik Rasbora tawarensis (Pisces: Cyprinidae) eggs after short‐term cryopreservation

2020 ◽  
Vol 51 (4) ◽  
pp. 1700-1705
Author(s):  
Zainal A. Muchlisin ◽  
Putri I. Sarah ◽  
Dhea F. Aldila ◽  
Kartini Eriani ◽  
Iwan Hasri ◽  
...  
2017 ◽  
Vol 29 (1) ◽  
pp. 191
Author(s):  
M. A. Bopape ◽  
T. L. Nedambale ◽  
C. M. Pilane ◽  
K. C. Lehloenya

Egg yolk is a common constituent of semen extender and protects sperm against cold shock. Besides its protective characteristic, chicken egg yolk is mostly included in extenders for semen preservation due to its abundant availability. The aim of the study was to compare egg yolk sources from different indigenous chicken egg yolk sources and storage temperatures on short-term preservation of South African indigenous goat semen. Semen was collected from 8 South African indigenous goats with artificial vagina during the breeding season (autumn). From each of the 8 goats, 6 replicates were done. Semen samples were randomly allocated into 4 treatment groups of Tris-based extenders containing 20% of egg yolk from White Leghorn as a control, Ovambo, Potchefstroom Koekoek, and Venda chicken breeds. The extended semen samples were stored at 5 or 25°C for 48 h. Semen samples were evaluated for sperm motility using computer-aided sperm analysis sperm viability and morphology with fluorescence microscope at 0, 3, 24, and 48 h. Data was analysed with ANOVA using Stata® version 12 (StataCorp, College Station, TX, USA) statistical software to test the differences between the treatments. The total, progressive, and rapid sperm motility rates were higher in freshly collected semen (90.6, 42.7, and 33.0%, respectively) compared with treatment groups. Semen extended with Tris without egg yolk had higher total sperm motility rate at both 5°C (48.1; 51.1%) and 25°C (43.3; 53.3%) temperatures for 48 h. Semen extended with egg yolk from different egg yolk sources had no sperm motility from 24 h when stored at 25°C. Semen extended with Tris without egg yolk (48.1%) had higher sperm motility than Ovambo, Potchefstroom Koekoek, Venda, and White Leghorn (3.7, 0, 0, and 0.4%, respectively) when stored at 5°C for 48 h. However, sperm motility declined when storage increased. In conclusion, addition of egg yolk had no effect on preserving goat semen. However, Tris-based extender without addition of egg yolk preserved semen for longer period than semen extended with egg yolk regardless of egg yolk origin or chicken breed.


Author(s):  
Kartini Eriani ◽  
Mustaqim Mustaqim ◽  
Iwan Hasri ◽  
R. Amalia ◽  
Al Azhar ◽  
...  

Background: Sperm cells are susceptible to oxidative stress during cryopreservation. Therefore, an antioxidant is necessary to protect them from damages. Fetal bovine serum (FBS) is one of potent antioxidants for fish sperm cryopreservation. Hence, the aims of this study are to examine the effect of FBS on sperm quality after a short period and to determine its optimum concentration on depik (Rasbora tawarensis). Methods: Depik fish were obtained from the Fish hatchery of Lukup Badak, Aceh Tengah District, Indonesia. Sperms collected from the fish were diluted in Ringer extenders containing FBS concentration of 10% (P1), 20% (P2), 30% (P3), 40% (P4), 50% (P5) and 60% (P6), filled into 2 ml cryotubes and equilibrated prior immersed into liquid nitrogen for 15 days. The parameters observed were sperm motility, consistency, pH, fertilization and hatching rates and DNA fragmentation post-thawing. Result: The ANOVA test indicates that the application of FBS in Ringer had a significant effect on sperm motility, fertilization and hatching rates (P less than 0.05). The highest motility (58.33%) was recorded at FBS 60% and significantly different from those at other concentrations. The laddering analysis showed that applying FBS protected the integrity of depik sperms DNA. It is concluded that the optimum concentration of FBS on depik sperm led to a short-term cryopreservation of 60%.


1976 ◽  
Vol 36 (01) ◽  
pp. 221-229 ◽  
Author(s):  
Charles A. Schiffer ◽  
Caroline L. Whitaker ◽  
Morton Schmukler ◽  
Joseph Aisner ◽  
Steven L. Hilbert

SummaryAlthough dimethyl sulfoxide (DMSO) has been used extensively as a cryopreservative for platelets there are few studies dealing with the effect of DMSO on platelet function. Using techniques similar to those employed in platelet cryopreservation platelets were incubated with final concentrations of 2-10% DMSO at 25° C. After exposure to 5 and 10% DMSO platelets remained discoid and electron micrographs revealed no structural abnormalities. There was no significant change in platelet count. In terms of injury to platelet membranes, there was no increased availability of platelet factor-3 or leakage of nucleotides, 5 hydroxytryptamine (5HT) or glycosidases with final DMSO concentrations of 2.5, 5 and 10% DMSO. Thrombin stimulated nucleotide and 5HT release was reduced by 10% DMSO. Impairment of thrombin induced glycosidase release was noted at lower DMSO concentrations and was dose related. Similarly, aggregation to ADP was progressively impaired at DMSO concentrations from 1-5% and was dose related. After the platelets exposed to DMSO were washed, however, aggregation and release returned to control values. Platelet aggregation by epinephrine was also inhibited by DMSO and this could not be corrected by washing the platelets. DMSO-plasma solutions are hypertonic but only minimal increases in platelet volume (at 10% DMSO) could be detected. Shrinkage of platelets was seen with hypertonic solutions of sodium chloride or sucrose suggesting that the rapid transmembrane passage of DMSO prevented significant shifts of water. These studies demonstrate that there are minimal irreversible alterations in in vitro platelet function after short-term exposure to DMSO.


2019 ◽  
Vol 6 (1) ◽  
pp. 1
Author(s):  
Muhammad Riyadhi ◽  
Anis Wahdi ◽  
Muhammad Rizal

ABSTRAK                                                                        Penelitian bertujuan untuk mengetahui efektivitas nira aren sebagai pengencer alternatif dalam proses pembekuan (kriopreservasi) semen kambing boer.Kriopreservasi semen kambing boer menggunakan pengencer tris-gliserol-kuning telur (P1 73-7-20%), nira aren-gliseol-kuning telur(masing-masing P2 74-6-20%, P3 73-7-20%, dan P4 72-8-20%) dan andromed (P5 tanpa mengandung kuning telur dan gliserol). Parameter evaluasi meliputi motilitas, viabilitas, dan membrane plasma utuh setelah pengenceran, ekuilibrasi dan thawing.  Evaluasi motilitas pasca thawing menunjukkan P5 52% berbeda nyata (P<0.05) dengan P1 42%, selanjutnya P5 dan P1 berbeda sangat nyata (P<0.05) dengan P2 8%, P3 6% dan P4 12%.  Viabilitas pasca thawing menunjukkan P5 65,4% tidak berbeda nyata (P>0,05) dengan P1 61,8%, akan tetapi P5 dan P1 berbeda sangat nyata (P<0.05) dengan P2 26,2%, P3 29,8%, dan P4 34%.  Membran plasma utuh (MPU) pasca thawing menunjukkan P5 66,2% tidak berbeda nyata (P>0,05) dengan P1 65,4%, akan tetapi keduanya berbeda sangat nyata (P<0.05) dengan P2 39%, P3 38%, dan P4 36,2%.  Disimpulkan kriopreservasi semen kambing boer dengan pengencer nira aren dan gliserol pada konsentrasi berbeda belum dapat dipergunakan sebagai sumber bibit berdasarkan standar nasional Indonesia.Kata Kunci : Kambing boer, semen, nira arenABSTRACTThe experiment was conducted to determine the effect of sugar palm juice as alternative extender for cryopreservation process of boer semen.Tris-glycerol-egg yolk (P1 73-7-20%), Sugar palm juice-glyserol-egg yolk (P2 74-6-20%, P373-7-20%, dan P4 72-8-20%), and andromed (P5) used as a extender  in the cryopreservation process of boer semen.  Sperm motility (%), live sperm (%) and sperm membrane integrity (%) were recorded after diluted, equilibration and freeze-thawing.  Result of post thawing motility showed that P5 52% was significantly different (P <0.05) with P1 42%, then P5 and P1 were significantly different (P <0.05) with P2 8%, P3 6% and P4 12%. Viability after thawing showed P5 65.4% was not significantly different (P> 0.05) with P1 61.8%, but P5 and P1 significantly different (P <0.05) with P2 26.2%, P3 29.8 %, and P4 34%. Spermmembrane integrity post-thawing showed P5 66.2% was not significantly different (P> 0.05) with P1 65.4%, but both were very significantly different (P <0.05) with P2 39%, P3 38% and P4 36.2%. Conclusions, sugar palm juice-glycerol-egg yolk with differentconcentrationsineffectively as an alternative extenderin cryopreservation of boer semen.Keywords: boer goat, semen, sugar palm juice


2016 ◽  
Vol 28 (7) ◽  
pp. 1004 ◽  
Author(s):  
Juliana C. Borges-Silva ◽  
Márcio R. Silva ◽  
Daniel B. Marinho ◽  
Eriklis Nogueira ◽  
Deiler C. Sampaio ◽  
...  

This study evaluated the use of cooled semen in a fixed-time artificial insemination (FTAI) program compared with frozen–thawed semen to improve pregnancy rates in beef cattle. Ejaculates of three bulls were collected and divided into two treatments: (1) frozen–thawed semen and (2) cooled semen. Egg-yolk extender without glycerol was used for the cooled semen treatment. Straws (25 × 106 spermatozoa) were submitted to cooling for preservation at 5°C for 24 h, after which FTAI was performed. Nelore cows (n = 838) submitted to FTAI were randomly inseminated using frozen–thawed semen or cooled semen. There was a 20% increase in the pregnancy per AI (P AI–1) using cooled semen compared with frozen–thawed semen (59.9 ± 4.7 vs 49.4 ± 5.0%; P < 0.005). There was no difference in P AI–1 among the bulls (P = 0.40). The frozen–thawed semen had fewer functional spermatozoa than did the cooled semen when evaluated by sperm motility (61.7 vs 81.0%), slow thermoresistance test (41.7 vs 66.7%) and hypoosmotic swelling test (38.3 vs 53.7%; P < 0.05). The percentage of sperm abnormalities did not differ between the freeze–thawing and cooling processes (18.6 vs 22.1%; P > 0.05). Because there was less damage to spermatozoa and improvement in P AI–1, the use of cooled semen instead of frozen–thawed semen is an interesting approach to increase reproductive efficiency in cattle submitted to a FTAI protocol.


2016 ◽  
Vol 47 (2) ◽  
pp. 60-67
Author(s):  
P. Folková ◽  
J. Šichtař ◽  
O. Šimoník ◽  
A. Dokoupilová ◽  
R. Rajmon

Abstract The aim of the study was to evaluate the effect of repeated semen collection and the substitution of normal egg yolk with clarified egg yolk to commercially produced semen extender on qualitative parameters of frozen-thawed canine semen. Two semen collections were scheduled in a 24-hour interval and in each of six dogs, three 1st and three 2nd collections were performed. The frozen-thawed sperm samples were prepared either with clarified or normal egg yolk and motility and viability were evaluated. The effect of the sequence of semen collection was demonstrated by significant differences in motility and also in viability of sperms both in native and frozen-thawed ejaculate. The percentage of viable sperms was significantly higher in samples from the 2nd compared to the 1st collection. This trend was the same also in motility except in native ejaculate. The addition of clarified egg yolk was beneficial for higher survival of sperms immediately after thawing and also after 30 min of incubation, compared to samples with normal egg yolk. Sperm motility evaluated after thawing was higher in samples with clarified egg yolk, without an apparent connection with semen collection sequence. The decrease of values of the qualitative parameters of sperms observed in the period of 30 min of incubation was significantly slowed down when clarified egg yolk was used. This was especially obvious in samples from the 2nd collection.


2013 ◽  
Vol 25 (8) ◽  
pp. 1185 ◽  
Author(s):  
M. Alvarez-Rodríguez ◽  
M. Alvarez ◽  
L. Anel-López ◽  
C. Martínez-Rodríguez ◽  
F. Martínez-Pastor ◽  
...  

Egg yolk low-density lipoproteins (LDL) and soybean lecithin were evaluated as replacements for egg yolk in extenders used for the cryopreservation of brown-bear spermatozoa. The motility, viability and acrosomal status of post-thawed spermatozoa were analysed, and an egg-yolk extender was used as a control. The total antioxidant capacity of these extenders was tested. Soybean lecithin showed an effect that was dependent on the soybean concentration (2%, 3.5% and 5%) and source (Type A: 24% l-α-phosphatidylcholine, and Type B: 14–23% l-α-phosphatidylcholine). Only semen cryopreserved with 5% Type A soybean exhibited a sperm motility similar to that of semen cryopreserved in egg-yolk-based extender after thawing, although the sperm viability and acrosome status were not as high. Semen frozen in an extender containing LDL (10–15%) exhibited improved sperm viability in comparison with the control, but sperm motility was lower. The LDL-based extender exhibited a higher anti-oxidant activity than the egg-yolk extender and soy lecithin-based extenders. The extenders with higher anti-oxidant activity showed improvements in frozen sperm viability but lower semen motility. These results indicate that soybean lecithin did not have the same protective effect as egg yolk during the freezing of brown-bear spermatozoa but suggest that LDL (10–15%) could be a useful substitute for egg yolk in these extenders.


2016 ◽  
Vol 37 (3) ◽  
pp. 1667 ◽  
Author(s):  
Robie Allan Bombardelli ◽  
Giovano Neumann ◽  
Cesar Pereira Rebechi Toledo ◽  
Eduardo Antônio Sanches ◽  
Denise Nascimento de Bastos ◽  
...  

The objective of this study was to evaluate the effect of copper-contaminated water on sperm motility, fertilization, and embryonic and larval development of silver catfish (Rhamdia quelen). A randomized experimental design with five treatments and four replicates was used. Two experiments were carried out: (1) controlled fertilization was performed under different levels of copper contamination and egg hatching was performed in clean water; and (2) copper-contaminated water was used for both fertilization and hatching assays. The time of sperm motility and sperm motility rates linearly decreased with increasing copper concentration in the water. Fertilization and hatching rates were also affected when the concentrations of copper in the water were above 0.0979 mg Cu+2 L-1 and 0.0331 mg Cu+2 L-1, respectively. Gamete exposure to levels between 15 mg Cu+2 L-1 and 60 mg Cu+2 L-1 for short periods of time negatively affected sperm motility, oocyte fertilization, and egg hatching rates. In addition, when gametes and embryos were exposed at levels above 0.03 mg Cu+2 L-1 during long periods of time, egg hatching rates were reduced, and at levels between 0.05 mg Cu+2 L-1 and 0.20 mg Cu+2 L-1 the number of abnormal larvae increased.


2006 ◽  
Vol 18 (2) ◽  
pp. 249
Author(s):  
G. Magarey ◽  
J. Herrick ◽  
K. Thiangtum ◽  
W. Tunwattana ◽  
W. Swanson

Wild populations of fishing cats (Prionailurus viverrinus) in Southeast Asia are in decline, primarily due to habitat loss. Because the fishing cat population in North American zoos is small (n = 69) and inbred (F = 0.17) with relatively low genetic variation (86%), infusion of new founder genes from Asia is a conservation priority. Importation of cryopreserved semen for use with IVF and ET may offer one alternative to the international transport of living animals. In this study, our objectives were to (1) compare motility longevity of fresh vs. frozen-thawed fishing cat spermatozoa in two culture media, (2) evaluate ovarian responses to exogenous gonadotropins, and (3) assess development of IVF embryos produced with fresh vs. frozen-thawed spermatozoa. Raw semen was collected via electroejaculation from male fishing cats (n = 4), divided into groups, and washed. Two sperm pellets were resuspended in either Ham's F10 medium (HF10; with 5% FBS) or our feline optimized culture medium (FOCM; with 0.4% BSA); another pellet was diluted in TEST egg yolk, cooled to 5�C over 3 h, glycerated (4%), and cryopreserved in straws over LN2 vapor. Frozen sperm samples were thawed, washed, and diluted in either HF10 or FOCM. Fresh and frozen-thawed sperm motility (percent motile, rate of forward progress) in each medium (10 � 106 motile sperm/mL) was assessed (at 0, 1, 3, and 6 h) in microdrops under oil during culture (38�C; 6% CO2 in air). Female fishing cats (n = 10) were treated with exogenous gonadotropins (150 IU eCG, 100 IU hCG, 85-h interval) and ovarian follicles were aspirated laparoscopically. Recovered oocytes were inseminated with fresh (2 � 105 motile sperm/mL) or frozen-thawed (5 � 105 motile sperm/mL) spermatozoa in FOCM microdrops; resulting embryos were either cryopreserved or cultured in FOCM (with 5% FBS added at 72 h post-insemination) for 7 days. Sperm motility over time did not differ (P > 0.05) between media for either fresh or frozen-thawed samples; however, across media, frozen-thawed sperm motility was lower (P < 0.05) and declined faster (P < 0.05) compared to fresh spermatozoa. Females produced an average (�SEM) of 9.8 � 2.9 mature ovarian follicles, allowing recovery of 7.3 � 2.6 high-quality oocytes per female. Oocyte cleavage percentage at 42 h p.i. was lower (P < 0.05) with frozen-thawed spermatozoa (38%, 11/29) compared to freshly collected spermatozoa (68%, 17/25). Overall, 35% (6/17) of cultured embryos developed to blastocysts with no difference (P > 0.05) between embryos produced with frozen-thawed (4/11) vs. fresh (2/6) spermatozoa. Although fishing cat sperm motility and fertility appear compromised after cryopreservation, our results demonstrate the ability of frozen-thawed spermatozoa to produce IVF embryos that are capable of developing to blastocyst stage in vitro. This work was supported by (NIH RR015388).


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