scholarly journals Effect of cryoprotectant on the motility, viability, fertilization, and DNA integrity of naleh fish Barbonymus sp. (Cyprinidae) sperm

2021 ◽  
Vol 58 ◽  
pp. e168702
Author(s):  
Siti Maulida ◽  
Kartini Eriani ◽  
Firman Muhammad Nur ◽  
Nur Fadli ◽  
Agung Setia Batubara ◽  
...  
Keyword(s):  
Ethylene Glycol ◽  
Protective Effect ◽  
Egg Yolk ◽  
Dna Integrity ◽  
Similar Concentration ◽  
Breeding Programs ◽  
Ringer’S Solution ◽  
Cryopreserved Sperm ◽  
Cryopreservation Method ◽  
Egg Fertility

Naleh fish Barbonymus sp. is a commercial freshwater fish, which is indigenous to Aceh, Indonesia. The population of this species has declined over the years as a result of habitat perturbations and overfishing. Hence, the crucial need to develop a cryopreservation method to support breeding programs. This involved the use of a cryoprotectant as an important component. The objective of this study, therefore, was to explore the best cryoprotectant for naleh fish spermatozoa, and a total of five types were tested. These include the DMSO, Methanol, Ethanol, Glycerol, and Ethylene Glycol at a similar concentration of 10%, which were individually combined with 15% egg yolk, and every treatment was performed in three replications. Conversely, Ringer’s solution was adopted as an extender, and the sperm was cryopreserved in liquid nitrogen for 15 days. The results showed significant influence on sperm motility and viability, as well as egg fertility of naleh fish (P <0.05), although the DMSO provided the best outcome, compared to others at 47.17%, 50.13%, and 45.67%, respectively. Furthermore, DNA fragmentation had not occurred in the fresh and cryopreserved sperm samples, indicating the protective effect of tested cryoprotectants. It is concluded that the 10% DMSO and 15% egg yolk is the best cryoprotectant for naleh fish spermatozoa.

scholarly journals Vital Analysis of Cryopreserved Sperm of Marbled Flounder, Pseudopleuronectes yokohamae

2021 ◽  
Vol 12 ◽  
Author(s):  
Shaharior Hossen ◽  
Soo Cheol Kim ◽  
Yusin Cho ◽  
Kang Hee Kho
Keyword(s):  
Sucrose Solution ◽  
Membrane Integrity ◽  
Dna Integrity ◽  
Ringer’S Solution ◽  
Sperm Dna ◽  
Significant Difference ◽  
Pseudopleuronectes Yokohamae ◽  
Cryopreserved Sperm ◽  
Motility Of Sperm ◽  
Cryopreservation Protocol

The marbled flounder (Pseudopleuronectes yokohamae) is a commercial flatfish in East Asia. The aim of this study was to improve its sperm cryopreservation protocol based on the vitality assessment of 7-day and 1-year cryopreserved sperm. Four extenders (extender-1: sucrose solution; extender-2: glucose solution; extender-3: fish Ringer's solution; and extender-4: modified fish Ringer's solution) were tested with a combination of five cryoprotectants (CPAs) (dimethyl sulfoxide: Me2SO; glycerol: GLY; ethylene glycol: EG; propylene glycol: PG; and methanol: MeOH) at four different concentrations (5, 10, 12, and 15%). Fluorescent technique was applied to detect the plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), and DNA integrity of fresh and cryopreserved sperm specimens. Fresh sperm was diluted at a ratio of 1:2 (sperm:extender). Post-thaw motility of sperm cryopreserved using 15% Me2SO along with either extender-1 (86.0 ± 5.2%) or extender-2 (85.7 ± 7.1%) was similar (p &gt; 0.05) to that of fresh sperm. Sperm cryopreserved using 12% GLY combined with extender-1 (83.67 ± 6.7%) or extender-2 (83.3 ± 4.7%) showed a similar motility to those cryopreserved with 15% Me2SO, but significantly lower from fresh sperm. The type of straw (0.25 or 0.50 mL) did not show any significant difference (p &gt; 0.05) in post-thaw sperm motility. The highest values of PMI and MMP were observed for 7-day cryopreserved sperm using extender-1 in combination with 15% Me2SO (91.0 ± 2.9% and 90.0 ± 2.0%, respectively) or 12% GLY (90.0 ± 1.3% and 90.0 ± 4.6%, respectively). These results were similar to those of fresh sperm (95.3 ± 2.1% and 92.9 ± 2.5%, respectively). PMI and MMP of 1-year cryopreserved sperm using extender-1 in combination with 15% Me2SO (90.3 ± 2.5% and 89.3 ± 2.1%, respectively) or 12% GLY (90.0 ± 4.4% and 88.7 ± 2.2%, respectively) were significantly similar (p &gt; 0.05) to those of fresh sperm. Sperm DNA integrity did not reveal any significant difference (p &gt; 0.05) between fresh and cryopreserved (7-day and 1-year) sperm. Based on the assessed sperm vitality indicators, a cryopreservation protocol using extender-1 in combination with 15% Me2SO or 12% GLY has potential for hatchery as well as to create a germplasm bank.

Protective effect of the hydro-alcoholic extract of Rubia cordifolia roots against ethylene glycol induced urolithiasis in rats

2010 ◽  
Vol 48 (4) ◽  
pp. 1013-1018 ◽  
Author(s):  
Kalyani Divakar ◽  
A.T. Pawar ◽  
S.B. Chandrasekhar ◽  
S.B. Dighe ◽  
Goli Divakar
Keyword(s):  
Ethylene Glycol ◽  
Protective Effect ◽  
Alcoholic Extract ◽  
Rubia Cordifolia

Comparing ethylene glycol with glycerol for cryopreservation of canine semen in egg-yolk TRIS extenders

2006 ◽  
Vol 66 (9) ◽  
pp. 2047-2055 ◽  
Author(s):  
Ana Martins-Bessa ◽  
António Rocha ◽  
A. Mayenco-Aguirre
Keyword(s):  
Ethylene Glycol ◽  
Egg Yolk

Protective Effect of Sucrose and Antioxidants on Cryopreservation of Sperm Motility and DNA Integrity in C57BL/6 Mice

2018 ◽  
Vol 16 (6) ◽  
pp. 444-450 ◽  
Author(s):  
Xiaowei Shi ◽  
Huanhuan Hu ◽  
Guojie Ji ◽  
Jing Zhang ◽  
Rui Liu ◽  
...  
Keyword(s):  
Protective Effect ◽  
Sperm Motility ◽  
Dna Integrity

97 IMPROVED CRYOPRESERVATION OF DOMESTIC CAT SPERMATOZOA IN A SOY LECITHIN-BASED EXTENDER

2011 ◽  
Vol 23 (1) ◽  
pp. 153 ◽  
Author(s):  
M. M. Vick ◽  
H. L. Bateman ◽  
W. F. Swanson
Keyword(s):  
Sperm Motility ◽  
Room Temperature ◽  
Egg Yolk ◽  
Tris Buffer ◽  
Domestic Cat ◽  
Dna Integrity ◽  
Freezing And Thawing ◽  
Soy Lecithin ◽  
Acridine Orange Staining ◽  
Acrosome Integrity

Development of a chemically defined, plant-based cryopreservation media would reduce extender variability and the potential for transmission of zoonotic pathogens compared with traditional egg-yolk-based extenders. The objective of this study was to compare effects of egg yolk- and soy lecithin-based cryopreservation media and the temperature of glycerol addition on sperm parameters following freezing and thawing of domestic cat spermatozoa. Fresh semen was collected by manual stimulation on 3 separate occasions from 4 adult male cats. Each ejaculate was washed to remove seminal plasma, divided into 4 equal aliquots, and extended at room temperature in one of the following treatments: 1) TEST-egg yolk (Irvine Scientific Inc., Santa Ana, CA, USA) medium with 4% glycerol (EYG); 2) TEST-egg yolk, with 4% glycerol added after cooling to 5°C (EY); 3) TES-Tris buffer with soy-lecithin (1%) and 4% glycerol (SLG); and 4) TES-Tris buffer with 1% soy-lecithin, and 4% glycerol added after cooling to 5°C (SL). Sperm progressive motility (%) and rate of progressive movement (scale of 0–5) were evaluated at 0, 1, 3, 6, and 24 h post-thaw. Sperm capacitation (chlortetracycline staining), acrosome integrity (FITC-PNA staining), and DNA integrity (acridine orange staining) were assessed at 15 min post-thaw. Data were exponentially transformed to achieve normal distribution and then subjected to GLM analysis to determine effects of media and temperature of glycerol addition on sperm traits. At 0 and 1 h post-thaw, acrosome integrity, DNA integrity and % sperm motility did not differ (P > 0.05) among treatments. However, % sperm motility was greater in the soy-based media compared to egg yolk-based media at 3, 6, and 24 h post-thaw (Table 1; P < 0.05). A higher percentage of uncapacitated spermatozoa were present in soy-based compared to egg-yolk based cryopreservation media (63.9 ± 9.3 v. 51.2 ± 11.5, respectively; P < 0.05), regardless of temperature of glycerol addition. Finally, addition of glycerol at 5°C resulted in higher % sperm motility compared to room temperature at 6 and 24 h post-thaw in both medium types (Table 1; P < 0.05). Our results suggest that use of a chemically defined, soy-based medium improves long-term motility and capacitation status of frozen–thawed domestic cat spermatozoa compared with cryopreservation in a traditional egg yolk-based extender. Table 1.Motile spermatazoa and motility score at 3, 6, and 24 h

scholarly journals Slow freezing versus vitrification for the cryopreservation of zebrafish (Danio rerio) ovarian tissue

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Lis S. Marques ◽  
Ana A. N. Fossati ◽  
Rômulo B. Rodrigues ◽  
Helen T. Da Rosa ◽  
Aryele P. Izaguirry ◽  
...  
Keyword(s):  
Plasma Membranes ◽  
Ovarian Tissue ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Slow Freezing ◽  
Mitochondrial Activity ◽  
Trypan Blue Exclusion ◽  
Cell Membrane Integrity ◽  
Cell Plasma ◽  
Cryopreservation Method

Abstract The aim of the present study was to compare the efficiency of vitrification and slow freezing techniques for the cryopreservation of zebrafish ovarian tissue containing immature follicles. In Experiment 1, assessment of cell membrane integrity by trypan blue exclusion staining was used to select the best cryoprotectant solution for each cryopreservation method. Primary growth (PG) oocytes showed the best percentage of membrane integrity (63.5 ± 2.99%) when SF4 solution (2 M methanol + 0.1 M trehalose + 10% egg yolk solution) was employed. The vitrification solution, which presented the highest membrane integrity (V2; 1.5 M methanol + 5.5 M Me2SO + 0.5 M sucrose + 10% egg yolk solution) was selected for Experiment 2. Experiment 2 aimed to compare the vitrification and slow freezing techniques in the following parameters: morphology, oxidative stress, mitochondrial activity, and DNA damage. Frozen ovarian tissue showed higher ROS levels and lower mitochondrial activity than vitrified ovarian tissue. Ultrastructural observations of frozen PG oocytes showed rupture of the plasma membrane, loss of intracellular contents and a large number of damaged mitochondria, while vitrified PG oocytes had intact mitochondria and cell plasma membranes. We conclude that vitrification may be more effective than slow freezing for the cryopreservation of zebrafish ovarian tissue.

scholarly journals Cryopreservation of Sperm of Labeo rohita (Hamilton, 1822) and Its Use in Fertilization and Hatching of Eggs

2013 ◽  
Vol 22 (1-2) ◽  
pp. 123-137 ◽  
Author(s):  
MRI Sarder ◽  
AKMA Rahman ◽  
MS Samad ◽  
KMS Nazrul ◽  
MKJ Bhuiyan
Keyword(s):  
Labeo Rohita ◽  
Egg Yolk ◽  
Artificial Breeding ◽  
Cryopreserved Sperm ◽  
Hatching Rates

Cryopreservation is one of the widely accepted and useful methods for preservation of gamete especially the spermatozoa.  Experiments were conducted to develop the protocols for cryopreservation of sperm of Labeo rohita and to assess the effect of cryopreserved sperm on fertilization and hatching. Six extenders Alsever’s solution, egg-yolk citrate, urea egg-yolk, Kurokura-2, Ma and Mb and five cryoprotectants ethanol, methanol, DMSO, DMA and glycerol were employed. Diluents were prepared by mixing the cryoprotectants at 10% concentration of the extenders (% v/v). Milt and diluents were mixed at a ratio of 1:9 for Alsever’s solution and Kurokura-2; and 1:4 for urea egg-yolk, egg-yolk citrate, Ma and Mb. Egg-yolk citrate with 10% DMSO showed best performance producing 85 ± 1.6% post-thaw motility followed by 83±2% and 81±2.9% with Alsever’s solution and urea egg-yolk respectively. Other extenders did not produce satisfactory results. Milt was diluted at six ratios (1:2, 1:4, 1:7, 1:9, 1:15 and 1:20) and 1:4 dilution showed the highest post-thaw motility for egg-yolk citrate and urea egg-yolk and 1:9 for Alsever’s solution. Six cryoprotectant concentrations (5, 7, 10, 15, 20% and 30%) were investigated and 10% concentration produced the highest post-thaw motility. In breeding trials, sperm preserved with egg-yolk citrate plus DMSO as well as Alsever’s solution plus DMSO fertilized eggs and produced hatchlings. The fertilization and hatching rates were 57 ± 7% and 46.5 ± 3.5% for egg-yolk citrate, and 33.5 ± 3.5% and 27±3% for Alsever’s solution respectively. Fresh sperm yielded 76 ± 4% fertilization and 70.5 ± 5.5% hatching. The protocol developed through this study can be applied for long-term conservation of genetic materials of L. rohita and the cryopreserved sperm can be used in artificial breeding for generating new individuals.DOI: http://dx.doi.org/10.3329/pa.v22i1-2.16474 Progress. Agric. 22(1 & 2): 123-137, 2011

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