scholarly journals Effect of α-tocopherol supplementation in diluents on the motility, viability and plasma membrane integrity of Simmental bull spermatozoa after cooling

2020 ◽  
Vol 9 (1) ◽  
pp. 1
Author(s):  
Sarah Azura ◽  
Hermin Ratnani ◽  
Suherni Susilowati ◽  
Mas'ud Hariadi ◽  
Abdul Samik ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Integrity ◽  
Skim Milk ◽  
Egg Yolk ◽  
Oxygen Species ◽  
Cold Temperatures ◽  
Plasma Membrane Integrity ◽  
Semen Storage ◽  
Simmental Cattle ◽  
Antioxidant Supplement

Semen storage in cold temperatures might cause an increase in reactive oxygen species (ROS) production. This condition resulted in spermatozoa damage and quality decrease. This study was conducted to investigate the effects of α-tocopherol supplementation in diluents on the motility, viability, and plasma membrane integrity of Simmental bull spermatozoa after cooling. Semen samples were diluted in skim milk egg yolk supplemented with 0, 0.5, 1.0, and 1.5 mM α-tocopherol respectively for control, Tl, T2, and T3. Spermatozoa were evaluated for their motility, viability, and membrane integrity in cooling temperature (5°C). The daily evaluation showed that 1.5 mM α-tocopherol was the best in maintaining motility, viability, and plasma membrane integrity, while 1.0 mM α-tocopherol was only good for maintaining viability. Therefore, it can be concluded that α-tocopherol at the concentration of 1.5 mM was an efficient antioxidant supplement for Simmental cattle semen in skim milk egg yolk diluent.

12 FERTILITY OF FROZEN EQUINE SPERM IN SYSTEMS FOR CRYOPRESERVATION

2012 ◽  
Vol 24 (1) ◽  
pp. 117
Author(s):  
R. R. D. Maziero ◽  
P. N. Guasti ◽  
I. D. P. Blanco ◽  
I. Martin ◽  
G. A. Monteiro ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Liquid Nitrogen ◽  
Membrane Integrity ◽  
Skim Milk ◽  
Egg Yolk ◽  
Automated System ◽  
Epifluorescence Microscopy ◽  
Computer Assisted ◽  
Plasma Membrane Integrity ◽  
Sperm Analysis

Optimizing cryopreservation of equine sperm will facilitate genetic banking and propagation of important horse strains through assisted reproduction. This study aimed to evaluate the motility pattern using computer-assisted sperm analysis (CASA) and plasma membrane integrity by epifluorescence microscopy of equine semen frozen in 0.5 mL straws at different freezing rates; also, a fertility trial was performed according to the freezing protocol. Three ejaculates from four stallions of various breeds (Mangalarga Marchador, Westfallen, Hanovarian and Arabian) and ages (5 to 20 years) were collected and processed for cryopreservation. The stallions were housed at the CERBEQ, Reproduction Centre of the Department of Animal Reproduction and Veterinary Radiology, UNESP. The ejaculates were filtered and submitted to analysis by CASA (HTM IVOS 12, Hamilton Thorne Research, USA). In addition, the plasma membrane integrity was determined by fluorescent probes. After evaluation, the ejaculates were diluted at 1:1 (extender:semen) with skim milk extender Botu-Semen™ and centrifuged at 600 × g for 10 min. The supernatant was removed and the pellet resuspended to a final concentration of 100 × 106 sperm mL–1 with milk-egg yolk freezing extender (Botu-Crio™). Semen was packaged in 0.5-mL straws (IMV, LAigle, France) and was placed in nitrogen for 20 min and then from room temperature to 5°C and then frozen in two different cooling systems: an isothermic box (42 cm × 28 cm × 12.5 cm) was placed upon racks suspended 6 cm above liquid nitrogen or other 20 min then immersed into nitrogen and automated system Mini Digitcool™ (IMV Technologies, France), cooling at a –40°C min–1 rate. All straws were stored in liquid nitrogen until thawing and analysis. The straws were thawed in a water bath at 46°C for 20 s and the samples were evaluated for progressive motility, angular progressive velocity, progressive velocity, track speed, percentage of rapid sperm and percentage of sperm with plasma membrane integrity. For the fertility trial, 65 clinically healthy mares had their oestrous cycle monitored by ultrasound and inseminated postovulation with sperm into the uterus. Ovulation was induced with 1 mL of deslorelin acetate (GnRH) injected IM when a 35-mm follicle was detected. Thirty-six hours later, mares were monitored every 6 h until ovulation was detected. When it was detected, mares were inseminated with 800 × 106 total sperm. Pregnancy was confirmed via ultrasound examination 15 days after ovulation. Pregnancy rate was 52.2% using the isothermic box and 60% using the automated machine. Statistical analysis from the frozen–thawed semen evaluated parameters was performed using the statistics software Proc. MIXED of SAS 9.1 and for the fertility trial, logistic regression using the Proc GENMOD from SAS 9.1. The conventional method using the isothermic box was similar to the automated machine with a fast freezing rate. Additionally, AI with 800 × 106 sperm frozen in the isothermic box or automated system resulted in similarly acceptable conception rates.

scholarly journals Donkey Epididymal Transport for Semen Cooling and Freezing

Animals ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 2209
Author(s):  
Yamilka Lago-Alvarez ◽  
Giorgia Podico ◽  
Lorenzo G. Segabinazzi ◽  
Lais L. Cunha ◽  
Leonardo Barbosa ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Mixed Models ◽  
Membrane Integrity ◽  
Skim Milk ◽  
Egg Yolk ◽  
Semen Parameters ◽  
Sperm Activation ◽  
Plasma Membrane Integrity ◽  
Semen Extender ◽  
Motility Parameters

The objectives of this study were to assess the cooling and freezing of donkey epididymal semen harvested immediately after castration (Experiment 1, n = 4) or after the shipment (24 or 48 h) of epididymides attached to testicles (Experiment 2, n = 14) or dissected apart (Experiment 3, n = 36). In each experiment, semen was frozen immediately (Non-Centrif) in an egg yolk-based semen extender (EY) or after processing through cushion-centrifugation (Centrif) while extended in a skim milk-based extender (SC). In all three experiments, cooled, pre-freeze, and post-thaw epididymal semen was assessed for total motility (TM), progressive motility (PM), plasma membrane integrity (PMI), and high mitochondrial membrane potential (HMMP). Data were analyzed with R using mixed models and Tukey’s test as posthoc. Results showed that the cooling of epididymal semen up to 24 h after harvesting did not affect motility parameters or plasma membrane integrity; furthermore, in Experiment 3, the post-thaw evaluation of both Centrif and Non-Centrif achieved similar TM and PM. Collectively, the post-thaw results revealed low motility parameters across groups; while, the PMI and HMMP did not reflect this trend, and the values remained high, suggesting that there was a lack of epididymal sperm activation with either centrifugation or extenders. In summary, freshly harvested and cooled-shipped and cooled semen had satisfactory semen parameters. Future studies need to address donkey epididymal semen fertility in mares and jennies.

scholarly journals Penambahan alfa-tokoferol dalam pengencer susu skim - kuning telur terhadap kualitas spermatozoa domba Sapudi yang disimpan pada suhu 5°C

2020 ◽  
Vol 9 (3) ◽  
pp. 69
Author(s):  
Satya Alysa Cahya Puspita ◽  
Suherni Susilowati ◽  
Trilas Sardjito ◽  
Abdul Samik ◽  
Indah Norma Triana ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Integrity ◽  
Skim Milk ◽  
Egg Yolk ◽  
Alpha Tocopherol ◽  
Control Treatment ◽  
Control Group ◽  
Cold Temperatures ◽  
Fresh Semen

Spermatozoa in fresh semen of Sapudi ram has a limited life span. The storage of semen in cold temperatures (5 °C) is intended to prolong the spermatozoa's life. However, storage in cold temperatures can lead to increased production of reactive oxygen species (ROS). This condition reduces the quality of spermatozoa. The purpose of this study was to determine the effect of alphatocopherol supplementation in skim milk-egg yolk extender on viability, motility, and plasma membrane integrity of Sapudi ram spermatozoa. Fresh semen derived from Sapudi ram was divided into four treatment groups. Control treatment (P0): semen was added in the extender of skim milkegg yolk without alpha-tocopherol. Three other treatments: P1, P2, and P3 semen were added in skim milk-egg yolk extender with the supplementation of 0.25, 0.5, and 1 gram alpha-tocopherol/ 100 mL extender, respectively. The results showed that the viability, motility, and integrity of the spermatozoa plasma membrane decreased gradually according to the storage length. Supplementation of skim milk-egg yolk extender with 0.5 gram of alpha-tocopherol/100 mL (P2) was able to maintain spermatozoa quality longer (p <0.05) than the control group. It can be concluded that alpha-tocopherol with a concentration of 0.5 g/100 mL of skim milk-egg yolk extender effectively maintains the quality of Sapudi ram spermatozoa in storage at 5 ° C.

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.

scholarly journals Accumulation of Non-Superoxide Anion Reactive Oxygen Species Mediates Nitrogen-Limited Alcoholic Fermentation by Saccharomyces cerevisiae

2010 ◽  
Vol 76 (24) ◽  
pp. 7918-7924 ◽  
Author(s):  
Ana Mendes-Ferreira ◽  
Belém Sampaio-Marques ◽  
Catarina Barbosa ◽  
Fernando Rodrigues ◽  
Vítor Costa ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Cell Cycle ◽  
Plasma Membrane ◽  
Superoxide Anion ◽  
Nitrogen Limitation ◽  
Alcoholic Fermentation ◽  
Membrane Integrity ◽  
Oxygen Species ◽  
Plasma Membrane Integrity ◽  
Cycle Arrest

ABSTRACT Throughout alcoholic fermentation, nitrogen depletion is one of the most important environmental stresses that can negatively affect the yeast metabolic activity and ultimately leads to fermentation arrest. Thus, the identification of the underlying effects and biomarkers of nitrogen limitation is valuable for controlling, and therefore optimizing, alcoholic fermentation. In this study, reactive oxygen species (ROS), plasma membrane integrity, and cell cycle were evaluated in a wine strain of Saccharomyces cerevisiae during alcoholic fermentation in nitrogen-limiting medium under anaerobic conditions. The results indicated that nitrogen limitation leads to an increase in ROS and that the superoxide anion is a minor component of the ROS, but there is increased activity of both Sod2p and Cta1p. Associated with these effects was a decrease in plasma membrane integrity and a persistent cell cycle arrest at G0/G1 phases. Moreover, under these conditions it appears that autophagy, evaluated by ATG8 expression, is induced, suggesting that this mechanism is essential for cell survival but does not prevent the cell cycle arrest observed in slow fermentation. Conversely, nitrogen refeeding allowed cells to reenter cell cycle by decreasing ROS generation and autophagy. Altogether, the results provide new insights on the understanding of wine fermentations under nitrogen-limiting conditions and further indicate that ROS accumulation, evaluated by the MitoTracker Red dye CM-H2XRos, and plasma membrane integrity could be useful as predictive markers of fermentation problems.

95 EFFECT OF CRYOPROTECTANT ADDITION AND REMOVAL AT 4°C ON MOTILITY AND PLASMA MEMBRANE INTEGRITY OF BOVINE CAUDAL EPDIDYMAL SPERMATOZOA

2006 ◽  
Vol 18 (2) ◽  
pp. 156
Author(s):  
C. Guerrero ◽  
S. Leibo ◽  
D. Paccamonti ◽  
B. Eilts ◽  
K. Bondioli ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Semen Analysis ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Single Step ◽  
Freezing Process ◽  
Computer Assisted ◽  
Chemical Toxicity ◽  
Plasma Membrane Integrity ◽  
Epididymal Spermatozoa

Cryopreservation of spermatozoa harvested from the epididymides would be a means of salvaging germplasm from genetically valuable males that die unexpectedly from injury, disease, or poaching. It is well known that the addition of cryoprotective agents (CPAs) is essential for sperm survival following the freezing process. However, CPAs can cause loss in sperm viability due to osmotic damage or chemical toxicity. The objective of this study was to determine the effects of single-step addition and/or removal of glycerol (GLY) or ethylene glycol (EG) on motility and plasma membrane integrity of bovine epididymal spermatozoa. Paired testes were obtained from mature bulls (n = 10) at a local abattoir and transported to the laboratory at 25–28°C within 4–6 h post-mortem. Epididymal spermatozoa were harvested by multiple incisions from the caudae epididymides of each bull, pooled, and washed in Brackett-Oliphant medium by centrifugation for 5 min at 500g. Pellets were resuspended in egg yolk-Tris-glucose-citric acid monohydrate medium (EYT-GC) at a concentration of 120 × 106 cells/mL and cooled to 4°C at a rate of 0.1°C/min. Specimens were allocated to each of five treatment groups: control (no CPA), 7% GLY, and 14% GLY, 7% EG, 14% EG. Then, replicate samples were diluted 1:1 in EYT-GC medium containing twice the final desired concentration of CPA. After being exposed for 10 min, each sample was diluted directly into EYT-GC at 4°C. Motility was assessed by means of a computer assisted semen analysis system and plasma membrane integrity was determined by SYBR 14 and propidium iodide staining followed by fluorescence microscopy. Differences among treatments were analyzed using one way ANOVA (P < 0.05). The results (Table 1) show that maximum survival, as assessed by measurements of motility and membrane integrity, was achieved with spermatozoa exposed to 7% EG. Almost identical intermediate levels of survival were observed with spermatozoa exposed to 7% GLY or 14% EG. The lowest survival was observed for spermatozoa exposed to 14% GLY. The results indicate that the use of EG as a cryoprotectant may minimize toxicity and osmotic damage to fresh bovine epididymal spermatozoa. Its efficacy as a CPA is currently being determined. Table 1. Sperm motility and membrane integrity (mean ± SEM) after addition of CPA to epididymal sperm

scholarly journals PENGARUH PENAMBAHAN EKSTRAK TEH HIJAU (Camellia sinensis) DALAM BAHAN PENGENCER SUSU SKIM KUNING TELUR TERHADAP KUALITAS SPERMATOZOA DOMBA SAPUDI YANG DISIMPAN PADA SUHU DINGIN

2020 ◽  
Vol 8 (2) ◽  
pp. 122
Author(s):  
Wahyu Retno Swari ◽  
Emy Koestanti Sabdoningrum ◽  
Wurlina Wurlina ◽  
Suherni Susilowati ◽  
Rochmah Kurnijasanti ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Green Tea ◽  
Plasma Membranes ◽  
Skim Milk ◽  
Egg Yolk ◽  
Green Tea Extract ◽  
Cold Temperatures ◽  
Tea Extract ◽  
Range Test

The purpose of this study was to determinated the effect and the best concentration of greentea extract in skim milk and egg yolk diluent for quality sapudi sheep spermatozoa were measured in motility, viability and intact plasma membrane that was stored on cold temperature. The semen was devided into four groups; skim milk and egg yolk diluent, 0,5% green tea extract in skim milk and egg yolk diluent, 0,1% green tea extract in skim milk and egg yolk diluent, and 0,15% green tea extract in skim milk and egg yolk diluent. Spermatozoa quality was observed day 1, day 2, day 3, day 4 and day 5 after being diluted. The data obtained was analyzed with the Analysis of Variant (ANOVA), followed by multiple range test duncan. The results showed that the highest percentage of motility, viability and intact plasma membrane derived from green tea extract 0,15% on skim milk and egg yolk diluent  for 1 day storage is 86.66b ± 2.58, 92.00b ± 1.78, and 74.16b ± 3.25. The lowest percentage of motility, viability and intact plasma membranes was obtained from the skim milk and egg yolk diluent for 5 days storage is 43.33a ± 6.05, 56.50a ± 3.08 and 28.33a ± 2.80. The conclusion of this study is the addition of 0.15% green tea extract in a skim milk and egg yolk diluent can maintain the quality of sperm for up to 5 days of storage at cold temperatures.

scholarly journals Quality of Sexed Sperm of Bali Bull in Regional Artificial Insemination Center of Riau Province

2019 ◽  
Vol 43 (2) ◽  
Author(s):  
Yendraliza Yendraliza ◽  
Anwar Efendi Harap ◽  
July Handoko ◽  
Muhammad Rodiallah
Keyword(s):  
Plasma Membrane ◽  
Pregnancy Rate ◽  
Artificial Insemination ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Female Offspring ◽  
Plasma Membrane Integrity ◽  
Frozen Semen ◽  
Calving Rate

This study aimed to evaluate the quality of frozen semen of Bali bull resulted from sexing procedure on calf or offspring production with desired sex. The tested sperm of Bali bull were collected from Bali bull raised at Regional Artificial Insemination Center of Riau Province (BIBD Riau). The study was carried out in 2 stages. The first stage was X and Y chromosome separation by albumin method. The extender used in the sexing procedure is trice citrate fructose and egg yolk. The second stage was mainly testing the sexed sperm collected in 60 Bali cow in Langkat Village, Bengkalis Regency. To determine the quality of post thawing frozen semen collected from the sexing procedure, the study evaluated motility, viability, mortality, abnormality and plasma membrane integrity of the spermatozoa. The pregnancy rate, calving rate, and birth accuracy of inseminated sexed sperm to offspring’ sex were also evaluated. The evaluation resulted in motility (66.3-75.3%), viability (70-78.5%), plasma membrane integrity (60-65.8%), abnormality (6.05-8.05%), mortality (20.05-30.05%), and pregnancy rate (83.33-90%). The calving rate on this study was 100% with the birth accuracy of 81.8% for male offspring and 40% for female offspring. As conclusion, the sexed sperm evaluated on this study have fairly good fertility.

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