109 Sperm cryopreservation in Eulamprus quoyii (Eastern water skink)

2019 ◽  
Vol 31 (1) ◽  
pp. 180
Author(s):  
R. Hobbs ◽  
L. Keogh ◽  
K. James ◽  
J. Baxter-Gilbert ◽  
M. Whiting
Keyword(s):  
Dimethyl Sulfoxide ◽  
Membrane Integrity ◽  
Adult Males ◽  
Sperm Cryopreservation ◽  
Treatment Groups ◽  
Dimethyl Acetamide ◽  
Rich Diversity ◽  
Sperm Volume ◽  
Semen Extender ◽  
Pooled Samples

Australia has a rich diversity and high endemism (93%) of reptilian species, the largest family being Scincidae (252 species), yet there continues to be a paucity of reports for gamete cryopreservation in reptilians (Clulow and Clulow 2016 Reprod. Fertil. Dev. 28, 1116-1132, DOI: 10.1071/RD15466). The goal of this study was to collect sperm from a locally abundant skink species (Eulamprus quoyii; Eastern water skink) to examine sperm sensitivity to cryopreservation. Wild-caught males (n=50; snout-vent length=103-126mm) were held in seminatural conditions for the duration of the study. Semen was collected during the breeding season (September-October) using an adapted ventral-massage technique (Molinia et al. 2010 Herp. Cons. Biol. 5, 311-319; retrieved from http://www.herpconbio.org). Sperm metrics (volume, concentration, progressive and total motility, and membrane integrity) were assessed subjectively under light or fluorescence microscopy. Results were compared using ANOVA. Sperm volume (9.43±5.18μL) and concentration (7.79×108±5.32×108 sperm/mL) did not correlate with snout-vent length in adult males (r2=0.025 and r2=0.15, respectively). Due to small volumes, sperm samples (motility >80%) from 4 to 6 males were pooled before allocation across treatments. For all cryopreservation experiments, sperm samples (5-10μL) were loaded into 0.2-mL French straws and frozen using a controlled-rate freezer (Cryobath; −6°C per minute), then plunged into LN. Sperm were thawed in a water bath at 35°C for 10s. In year 1, pooled samples (n=3) were maintained at room temperature (21±1.5°C) either raw, or diluted in PBS, Tris-citrate glucose, TLHepes, or Ham’s F-10 and assessed at 0, 1, 3, 16, 40h. Tris-citrate-glucose-diluted sperm had significantly lower total motility from 3h (36.7±11.5%; P<0.05), decreasing to 1% motility by 16h. Up to 70% motility could be maintained for 16h in all other treatment groups. In year 2, pooled samples (n=3) were cooled to 4°C over a period of 2h, then gradually diluted 1:1 with buffer to a final concentration of either 0.6, 1.35, or 2.7M cryoprotectant (CPA; dimethyl sulfoxide, dimethyl acetamide, glycerol) in PBS or no CPA. Sperm diluted in 1.35 and 2.7M dimethyl sulfoxide in PBS had significantly (P<0.05) higher kinetic rating and proportion of live sperm than control or dimethyl acetamide treatments; 1.35 and 2.7M glycerol were intermediate. In year 3, dilution and cryopreservation using 1.35M CPA in complex diluents, Tris-yolk buffer (20.1±2.6% live) and Beltsville poultry semen extender (29.7±2.0% live), did not significantly improve sperm survival compared to PBS (26.4±2.7% live); however, post-thaw progressive motility (5±1.1%) was significantly (P < 0.05) higher with 1.35M dimethyl sulfoxide Tris-yolk buffer than all other treatment groups. In conclusion, dimethyl sulfoxide yields promise for sperm cryopreservation in a skink, but further studies are required.

scholarly journals Effects of freezing and activation on membrane quality and DNA damage in Xenopus tropicalis and Xenopus laevis spermatozoa

2017 ◽  
Vol 29 (8) ◽  
pp. 1556 ◽  
Author(s):  
S. Morrow ◽  
J. Gosálvez ◽  
C. López-Fernández ◽  
F. Arroyo ◽  
W. V. Holt ◽  
...  
Keyword(s):  
Xenopus Laevis ◽  
Membrane Damage ◽  
Membrane Integrity ◽  
Subsequent Development ◽  
Xenopus Tropicalis ◽  
Model Organisms ◽  
Sperm Chromatin ◽  
Dna Integrity ◽  
Sperm Cryopreservation ◽  
Sperm Plasma Membrane

There is growing concern over the effect of sperm cryopreservation on DNA integrity and the subsequent development of offspring generated from this cryopreserved material. In the present study, membrane integrity and DNA stability of Xenopus laevis and Xenopus tropicalis spermatozoa were evaluated in response to cryopreservation with or without activation, a process that happens upon exposure to water to spermatozoa of some aquatic species. A dye exclusion assay revealed that sperm plasma membrane integrity in both species decreased after freezing, more so for X. laevis than X. tropicalis spermatozoa. The sperm chromatin dispersion (SCD) test showed that for both X. tropicalis and X. laevis, activated frozen spermatozoa produced the highest levels of DNA fragmentation compared with all fresh samples and frozen non-activated samples (P < 0.05). Understanding the nature of DNA and membrane damage that occurs in cryopreserved spermatozoa from Xenopus species represents the first step in exploiting these powerful model organisms to understand the developmental consequences of fertilising with cryopreservation-damaged spermatozoa.

Dimethylacetamide can be used as an alternative to glycerol for the successful cryopreservation of koala (Phascolarctos cinereus) spermatozoa

2008 ◽  
Vol 20 (6) ◽  
pp. 724 ◽  
Author(s):  
Yeng Peng Zee ◽  
William V. Holt ◽  
Jaime Gosalvez ◽  
Camryn D. Allen ◽  
Vere Nicolson ◽  
...  
Keyword(s):  
Disulfide Bonds ◽  
Membrane Integrity ◽  
Sperm Nucleus ◽  
Sperm Chromatin ◽  
Similar Degree ◽  
Sperm Cryopreservation ◽  
Phascolarctos Cinereus ◽  
Degree Of Protection ◽  
Fish Sperm ◽  
Cryopreservation Protocol

Swelling of koala sperm chromatin following cryopreservation has largely been attributed to the absence of intermolecular disulfide cross-linkages in the marsupial sperm nucleus. Fish spermatozoa also lack disulfide bonds within their chromatin, but have been successfully cryopreserved. The present study examined the hypothesis that the cryoprotectants used for fish sperm cryopreservation would confer a similar degree of protection on koala spermatozoa. Three concentrations each of five cryoprotectants (dimethyl sulfoxide, methanol, propylene glycol, ethylene glycol and dimethylacetamide (DMA)) were evaluated. Each treatment was compared against an established koala sperm cryopreservation protocol that uses 14% glycerol. Post-thaw assessment of progressive motility, plasma membrane integrity and mitochondrial membrane potential (MMP) revealed that protocols using 15% DMA achieved 62.2 ± 3.6% (P < 0.05) sperm survival, of which 79% (P < 0.05) had high MMP, an improvement of 32% and 40%, respectively, over sperm frozen in 14% glycerol. The percentage of spermatozoa with swollen nuclei was also lowest when frozen in 15% DMA, both immediately after thawing (18.0 ± 3.5%; P < 0.05) and after 2 h incubation at 35°C (35.8 ± 4.4%; P < 0.05). A second study was conducted to determine the optimal concentration of DMA for use in the cryopreservation of koala spermatozoa. High DMA concentrations (17.5% and 20%) resulted in significantly lower proportions of live spermatozoa showing high MMP immediately after thawing compared with spermatozoa frozen in the lower concentrations. The percentage of koala spermatozoa with swollen chromatin following cryopreservation was not affected by DMA concentration.

Glutathione Supplemented Soybean Lecithin Based Semen Extender Is Not Suitable for Buck Sperm Cryopreservation.

2012 ◽  
Vol 87 (Suppl_1) ◽  
pp. 420-420
Author(s):  
Hossein Salmani ◽  
Mahdi Zhandi ◽  
Malek Shakeri ◽  
Armin Towhidi ◽  
Ahmad Zare Shahneh ◽  
...  
Keyword(s):  
Soybean Lecithin ◽  
Sperm Cryopreservation ◽  
Semen Extender

scholarly journals The Effect of Adding Different Levels of Curcumin and Its Nanoparticles to Extender on Post-Thaw Quality of Cryopreserved Rabbit Sperm

Animals ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1508 ◽  
Author(s):  
Sameh Abdelnour ◽  
Mahmoud Hassan ◽  
Amer Mohammed ◽  
Ahmad Alhimaidi ◽  
Naif Al-Gabri ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Positive Influence ◽  
Control Group ◽  
Curcumin Nanoparticles ◽  
Total Antioxidant ◽  
Semen Extender ◽  
Apoptosis Process ◽  
Structural Levels

The cryopreservation process adversely affects sperm function and quality traits, causing some changes at biochemical and structural levels, due to mechanical, thermal, osmotic, and oxidative damage. Supplementation with curcumin nanoparticles could prevent and even revert this effect and could enhance the post/thawed sperm quality in the rabbit. The study amid to explore the effect of curcumin (CU) and curcumin nanoparticles (CUNPs) supplementation in semen extender on post/thawed rabbit sperm quality. Twelve fertile, healthy rabbit bucks were included, and the ejaculates were collected using artificial vaginas. Rabbit pooled semen was cryopreserved in tris-yolk fructose (TYF) extender without any supplement (control group) or extender supplemented with CU at levels of 0.5, 1 or 1.5 µg/mL (CU0.5, CU1.0, and CU1.5, respectively) or CUNPs at levels of 0.5, 1, 1.5 (CUNPs0.5, CUNPs1.0, and CUNPs1.5, respectively) and was packed in straws (0.25 mL) and stored in liquid nitrogen (−196 °C). Results revealed that CUNPs1.5 had a positive influence (p < 0.05) on post-thawing sperm progressive motility, viability, and membrane integrity as compared with the other groups. Percentages of dead sperm, abnormalities, early apoptotic, apoptotic, and necrotic sperm cells reduced (p < 0.05) in CUNPs1.5 as compared to other treatments. Using 1.5 µg/mL of CUNPs significantly improved total antioxidant capacity (TAC), GPx, while MDA and POC reduced (p < 0.05) in CU1.5 in comparison with other groups. SOD values were enhanced (p < 0.05) in CUNPs1.0 and CUNPs1.5 in relation with other treatments. Conclusively, the addition of curcumin and its nanoparticles to the extender can improve the post-thawed quality of rabbit sperm via redox signaling and reduce the apoptosis process.

scholarly journals Conditioned Medium from Canine Amniotic Membrane-Derived Mesenchymal Stem Cells Improved Dog Sperm Post-Thaw Quality-Related Parameters

Animals ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 1899
Author(s):  
Feriel Yasmine Mahiddine ◽  
Jin Wook Kim ◽  
Ahmad Yar Qamar ◽  
Jeong Chan Ra ◽  
Soo Hyun Kim ◽  
...  
Keyword(s):  
Kinetic Parameters ◽  
Conditioned Medium ◽  
Amniotic Membrane ◽  
Membrane Integrity ◽  
Mitochondrial Activity ◽  
Flow Cytometry Analysis ◽  
Sperm Cryopreservation ◽  
Control Groups ◽  
Freezing Medium ◽  
And Control

This study investigated the effects of conditioned medium (CM) from canine amniotic membrane-derived MSCs (cAMSCs) on dog sperm cryopreservation. For this purpose, flow cytometry analysis was performed to characterize cAMSCs. The CM prepared from cAMSCs was subjected to proteomic analysis for the identification of proteins present in the medium. Sperm samples were treated with freezing medium supplemented with 0%, 5%, 10%, and 15% of the CM, and kinetic parameters were evaluated after 4–6 h of chilling at 4 °C to select the best concentration before proceeding to cryopreservation. Quality-related parameters of frozen–thawed sperm were investigated, including motility; kinetic parameters; viability; integrity of the plasma membrane, chromatin, and acrosome; and mitochondrial activity. The results showed that 10% of the CM significantly enhanced motility, viability, mitochondrial activity, and membrane integrity (p < 0.05); however, the analysis of chromatin and acrosome integrity showed no significant differences between the treatment and control groups. Therefore, we concluded that the addition of 10% CM derived from cAMSC in the freezing medium protected dog sperm during the cryopreservation process.

Effect of platelet activating factor (PAF) supplementation in semen extender on viability and ATP content of cryopreserved canine spermatozoa

2010 ◽  
Vol 13 (4) ◽  
pp. 571-579 ◽  
Author(s):  
W. Kordan ◽  
M. Lecewicz ◽  
R. Strzeżek ◽  
A. Dziekońska ◽  
L. Fraser
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Mitochondrial Function ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Platelet Activating Factor ◽  
Computer Assisted ◽  
Atp Content ◽  
Plasma Membrane Integrity ◽  
Semen Extender

Effect of platelet activating factor (PAF) supplementation in semen extender on viability and ATP content of cryopreserved canine spermatozoa The aim of this study was to investigate the effect of platelet activating factor (PAF) on the quality characteristics of cryopreserved canine spermatozoa. Cryopreserved semen of 5 mixed-breed dogs was treated with different concentrations of exogenous PAF (1 × 10-3M, 1 × 10-4M, 1 × 10-5M and 1 × 10-6M) and examined at different time intervals (0, 30, 60 and 120 min). Cryopreserved semen treated without PAF was used as the control. Sperm quality was evaluated for motility (computer-assisted semen analysis, CASA), mitochondrial function (JC-1/PI assay) and plasma membrane integrity (SYBR-14/PI assay and Hoechst 33258). Also, ATP content of spermatozoa was determined using a bioluminescence assay. Treatment of cryopreserved semen with 1 × 10-3 M PAF at 120 min of incubation resulted in significantly higher total sperm motility compared with the control. It was observed that PAF-improved total sperm motility was concurrent with enhanced sperm motility patterns after treatment of cryopreserved semen. Treatment of cryopreserved semen with PAF did not improve either sperm mitochondrial function or plasma membrane integrity, as monitored by different fluorescent membrane markers. Furthermore, ATP content of cryopreserved spermatozoa was significantly higher when PAF was used at a concentration of 1 × 10-3 M compared with the control and other PAF treatments, regardless of the incubation time. The findings of this study indicated that treatment with 1 × 10-3 M PAF at 120 min of incubation rendered better quality of cryopreserved canine semen, which was associated with improved sperm motility parameters and ATP content. It can be suggested that exogenous PAF addition is beneficial as a supplement for canine semen extender used for.

Embryonic developmental plasticity in the long-snouted seahorse (Hippocampus reidi, Ginsburg 1933) in relation to parental preconception diet

2016 ◽  
Vol 28 (7) ◽  
pp. 1020 ◽  
Author(s):  
Francisco Otero-Ferrer ◽  
Marisol Izquierdo ◽  
Alireza Fazeli ◽  
William V. Holt
Keyword(s):  
Fatty Acid ◽  
Growth And Development ◽  
Developmental Plasticity ◽  
The Other ◽  
Adult Males ◽  
Fatty Acid Profiles ◽  
Commercial Diet ◽  
Treatment Groups ◽  
Offspring Growth ◽  
Males And Females

The aim of the present study was to investigate the hypothesis that parental periconception nutrition in adult seahorses affects the development and growth of their offspring. We tested the hypothesis that because seahorse embryos develop inside the male’s brood pouch, manipulation of the male’s diet would affect offspring growth and development independently of the female’s diet. Adult males and females were fed separately with either wild-caught crustaceans or commercial aquarium diet for 1 month before conception to influence the periconception environment. Approximately 10 000 offspring were obtained from four different treatment groups (Male/Wild or Male/Commercial × Female/Wild or Female/Commercial). Weights, physical dimensions and fatty acid profiles of the newborns were determined. Offspring produced when the males receiving commercial diet were mated with wild-fed females were larger (P < 0.05) than those produced by wild-fed males. When both males and females were fed with commercial diet, their offspring were significantly smaller than those from the other treatment groups. When commercial diet-fed females were mated with wild-fed males, the offspring showed distortion of the snout : head length ratio. These results support the view that the preconception diet received by males and females differentially affects embryonic development.

scholarly journals Effect of sunflower lecithin on Kalahari Red goat semen during cryopreservation

2018 ◽  
Vol 51 (1) ◽  
pp. 21-28 ◽  
Author(s):  
Sakirat Opeyemi Adeyanju ◽  
James Olatinbo Daramola ◽  
Jimoh Alao Olanite ◽  
Olufiropo Samson Awokola
Keyword(s):  
Soybean Lecithin ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Domestic Animal ◽  
Arginase Activity ◽  
Semen Parameters ◽  
Phospholipid Content ◽  
Acrosome Integrity ◽  
Semen Extender ◽  
Different Levels

Abstract Soybean lecithin had been used as an alternative to egg yolk in domestic animal semen extender during cryopreservation due to its characteristic phospholipid content which played a major cryoprotective role. This composition of soybean lecithin informed the replacement of soybean with sunflower lecithin (SL) in the extender for the Kalahari Red (KR) buck semen cryopreservation in this study. Effect of different levels of SL on the quality of the KR buck semen during cryopreservation using slow freezing method was evaluated. Semen samples were collected from four KR bucks of between two and two and half of age using artificial vagina, evaluated for motility and then diluted in extenders containing different levels of SL (1.5%, 3.0% and 4.5%) as experimental group and 0% SL or 20% egg yolk as control. Semen parameters including motility, acrosome integrity (AcI), membrane integrity (MI), malondialdehyde (MDA) concentration, cholesterol level and seminal arginase activity were evaluated for. The results showed that motility, acrosome integrity (AI) and membrane integrity were comparable at 0%, (22.00 ± 4.58, 82.00 ± 3.51 and 96.00 ± 2.03); 1.5%, (23.00 ± 2.08, 87.00 ± 3.79 and 89.00 ± 2.08); 3.0%, (13.00 ± 2.52, 81.33 ± 0.41 and 76.67 ± 1.20) and 4.5% (11.00 ± 4.51, 85.33 ± 9.88 and 84.00 ± 8.50), respectively, after thawing. SL at 0% had the highest (P < 0.05) values for MDA, cholesterol and seminal arginase activity (1.10 ± 0.008 nmol/ml, 236.35 ± 4.08 mg/dl and 0.54 ± 3.3 E-3 units/mg protein, respectively). Our data suggest that 1.5% sunflower lecithin can be used in place of soy lecithin as a substitute for egg yolk during the cryopreservation of caprine semen.

scholarly journals Cryopreservation of Indonesian native chicken semen by using dimethyl sulfoxide and various level of ethylene glycol as cryoprotectants

2020 ◽  
Vol 21 (12) ◽  
Author(s):  
KHAERUDDIN KHAERUDDIN ◽  
JUNAEDI JUNAEDI ◽  
HASTUTI HASTUTI
Keyword(s):  
Ethylene Glycol ◽  
Dimethyl Sulfoxide ◽  
Liquid Nitrogen ◽  
Egg Yolk ◽  
Sperm Cryopreservation ◽  
Randomized Design ◽  
Native Chicken ◽  
Ringer’S Lactate ◽  
Completely Randomized Design

Abstract. Khaeruddin, Junaedi, Hastuti. 2020. Cryopreservation of Indonesian native chicken semen by using dimethyl sulfoxide and various level of ethylene glycol as cryoprotectants. Biodiversitas 21: 5718-5722. Imported purebred chickens are becoming more popular and a regular staple in Indonesia. Therefore, it is necessary to strengthen conservation efforts to preserve Indonesian chickens, one of which is by means of sperm cryopreservation. This study aimed to determine the effects of the addition of DMSO and different concentrations of ethylene glycol to a Ringer’s lactate egg yolk (RLY)-or coconut water egg yolk (CWY)-based extender on the quality of frozen-thawed Indonesian chicken sperm. This study was used nine Indonesian native roosters about 20 months of age. The semen extenders used in this study were RLY + DMSO 7%, RLY + ethylene glycol 3%, RLY + ethylene glycol 5%, RLY + ethylene glycol 7%, CWY + DMSO 7%, CWY + ethylene glycol 3%, CWY + ethylene glycol 5% and CWY + ethylene glycol 7%. Liquid semen was packaged in 0.25 mL straw, then cooled at 5oC for 2 hours, frozen at 5 cm above liquid nitrogen for 10 minutes, following stored in a liquid nitrogen container for 24 hours. The semen straws were thawed at 37oC for 30 seconds. Statistical analysis for multiple comparisons was performed as a completely randomized design with eight treatment levels and seven replications. The results showed that there were no differences in sperm motility, recovery rate, and abnormality between extenders after the freeze-thaw process. Whereas, RLY + DMSO 7% was the highest sperm viability.

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