scholarly journals Glutathione supplementation in semen extender improves rabbit sperm attributes during refrigeration

2021 ◽  
Vol 29 (2) ◽  
pp. 81
Author(s):  
Ejaz Ahmad ◽  
Zahid Naseer ◽  
Melih Aksoy
Keyword(s):  
Sperm Motility ◽  
Acrosome Reaction ◽  
Reaction Rate ◽  
Sperm Quality ◽  
Final Concentration ◽  
Sperm Viability ◽  
Acrosome Integrity ◽  
Semen Extender ◽  
Sperm Motion ◽  
Artificial Vagina

In the present study, we evaluated the sustaining effect of various glutathione (GSH) concentrations in extender on rabbit sperm attributes during storage at 5°C for 24 h. Semen was collected from regular donor rabbit bucks using an artificial vagina and initially evaluated for sperm quality. The qualifying ejaculates were diluted with one of the extenders having 0, 1, 2, 4 or 8 mM GSH, to achieve a final concentration of 1×108 sperm/mL. The extended samples were stored at 5°C for 24 h. Sperm motility, motion kinetics, acrosome integrity and viability were assessed after 3, 6, 12 and 24 h of storage. The results showed that total sperm motility and sperm motion kinetics (oscillation index of the sperm, straightness index and beat cross frequency) were influenced (<em>P</em>&lt;0.05) by glutathione dose and refrigeration time. An interaction of (<em>P</em>&lt;0.05) GSH concentrations and refrigeration time was observed for sperm viability and acrosome reaction rate. In conclusion, the 4 mM GSH supplemented extender’s protective influence was remarkable to maintainrabbit sperm quality for 24 h 5°C.

scholarly journals Effect of Centrifugation on Motility, Sperm Capacitation and Acrosome Reaction in Soy Bean and Avocado Seed Milk Extenders of Cryopreserved Goat Spermatozoa

2017 ◽  
Vol 50 (1) ◽  
pp. 13-18
Author(s):  
James Ola Daramola
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Acrosome Reaction ◽  
Sperm Quality ◽  
West African ◽  
West African Dwarf ◽  
Artificial Vagina ◽  
Avocado Seed

Abstract Removal of seminal plasma by centrifugation (0 centrifugation, 1 centrifugation, 2 centrifugations, 3 centrifugations) and preservation in two different tris-extenders viz., avocado seed milk (ASM) and soy bean milk (SBM) based extenders were studied for their ability to support motility, in vitro capacitation and acrosome reaction of spermatozoa obtained from West African Dwarf (WAD) goat bucks during cryopreservation. Semen samples collected with the aid of artificial vagina were centrifuged for one, two and three times. The centrifuged samples were diluted with the two tris-extenders each containing 20 mL of avocado seed milk and soybean milk and cryopreserved for 30 days. The results showed higher (P < 0.05) sperm motility (P < 0.05) with increased centrifugation times. Spermatozoa that were centrifuged had higher (P < 0.05) percentage of acrosome reaction and capacitation with increased centrifugation times compared to the control. Optimal improvement in these parameters was obtained with increased centrifugation times. The findings revealed that removal of seminal plasma by centrifugation improved sperm quality of WAD goat bucks during cryopreservation and optimum improvement was achieved consistently with 3 centrifugations.

163 EVALUATION OF VIABILITY OF BULL SEMEN COLLECTED BY ELECTRO-EJACULATION USING COMMERCIAL SEMEN EXTENDER AND 2 CULTURE MEDIA AT CONTROLLED ROOM TEMPERATURE

2017 ◽  
Vol 29 (1) ◽  
pp. 190
Author(s):  
A. M. Raseona ◽  
O. A. Ajao ◽  
L. D. Nethengwe ◽  
L. R. Madzhie ◽  
T. L. Nedambale ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Assisted Reproductive Technologies ◽  
Culture Media ◽  
Reproductive Technologies ◽  
Sperm Viability ◽  
Progressive Motility ◽  
Bull Semen ◽  
Significant Difference ◽  
Semen Extender ◽  
Motility Rate

Preservation of semen is an important process to ensure that semen quality is sufficient for assisted reproductive technologies. The aim of this study was to evaluate the viability of bull semen collected by electro-ejaculation using commercial semen extender and 2 modified culture media stored at controlled RT (24°C) for 72 h. Two Nguni bulls were used for semen collection; after collection, the semen was evaluated macroscopically for volume, pH, and colour, and microscopically for sperm motility, viability, and morphology. Uncontaminated semen samples with progressive motility >70% and morphological defects <20% were pooled after collection before being aliquoted into 3 extenders, namely Triladyl, modified Ham’s F10, and TCM-199 culture media, at a dilution ratio of 1:4 and then stored at controlled RT (24°C). Sperm motility rate was analysed using the computer-aided sperm analyser after 0, 24, 48, and 72 h of storage. Sperm morphology and viability was performed after staining the sperm cells with spermac and nigrosine-eosin stain, respectively. The study was replicated 4 times and data were analysed using ANOVA. Triladyl had a higher sperm viability rate (41.3%) and total motility rate (96.3%) for 72 h (P < 0.01) compared with the 2 modified culture media, Ham’s F10 (26.5 and 86.8%) and TCM-199 (25.0 and 86.7%), respectively. However, Ham’s F10 had higher progressive motility rate (37.8%) as compared with the other extenders, TCM-199 (31.7%) and Triladyl (23.4). There was no significant difference (P > 0.05), in viability rate between Ham’s F10 (26.5%) and TCM-199 (25.0%). No significant difference (P > 0.05) in straight line velocity was observed for the three extenders. Furthermore, no significant difference was observed in total sperm abnormalities, except for reacted acrosomes and absent tails (P > 0.05), between the 2 Nguni bulls. Nguni semen can be preserved in Triladyl or modified Ham’s F10 and TCM-199 culture media stored at 24°C and stay viable for 72 h. Triladyl proved to be the best suitable extender of the 3 extenders, showing higher sperm viability and total motility rate as compared with Ham’s F10 and TCM-199 modified culture media.

scholarly journals Comparison of Extenders With the Addition of Egg Yolk for Cooling Alpaca Sperm Obtained From Deferent Ducts

2020 ◽  
Vol 7 ◽  
Author(s):  
Mariana Lucía Bertuzzi ◽  
Edita Yola Torres ◽  
Teodosio Huanca ◽  
Deborah Neild ◽  
María Ignacia Carretero
Keyword(s):  
Sperm Motility ◽  
Chromatin Condensation ◽  
Egg Yolk ◽  
Final Concentration ◽  
Sperm Parameters ◽  
Membrane Function ◽  
Progressive Motility ◽  
Evaluation Time ◽  
Viable Sperm ◽  
Acrosome Integrity

The use of non-commercial and commercial extenders for cooling alpaca sperm has already been reported, the latter showing certain advantages over the first. The Andromed® (AM) extender was created for use in ruminants and has also been tested in ejaculated and epididymal alpaca sperm. According to the manufacturer, this extender does not need the addition of egg yolk (EY); however, it is known that the addition of EY to some extenders improves the preservation of cooled sperm. The objective of this study therefore was to compare a non-commercial extender (Tris) with the addition of EY vs. the commercial extender AM with and without the addition of EY, for cooling alpaca sperm obtained from diverted deferent ducts. Fifteen pools of deferent duct sperm were formed using samples from two or three different males for each. Each sperm pool was evaluated and then divided into three aliquots that were diluted to a final concentration of 30 × 106 sperm ml-1 (0 h) with either: (1) Tris with 20% EY (T-EY), (2) AM, or (3) AM with 20% EY (AM-EY). Samples were cooled to 5°C and the following sperm parameters were evaluated after 24 and 48 h of storage: motility, viability, membrane function, acrosome integrity, morphology, and chromatin condensation. Motility was also evaluated after 72 h of storage. The samples that best preserved progressive and total sperm motility at the 24 and 48 h evaluation periods were the ones diluted with AM-EY, observing that with this extender these motility patterns decreased significantly after 72 h of storage compared to time 0 h (p &lt; 0.05). A significant decrease (p &lt; 0.05) in total and progressive motility was observed at 48 h for the T-EY and AM extender compared to 0 h. AM was the only extender in which the percentages of viable sperm decreased significantly (p &lt; 0.05) after 48 h of conservation. For the rest of sperm parameters evaluated, no significant differences were observed between any of the extenders at any evaluation time. The Andromed® extender with the addition of 20% EY could be an alternative option for cooling alpaca sperm obtained from deferent ducts.

Sperm viability and morphology of two genetically diverse Merino lines

2000 ◽  
Vol 12 (6) ◽  
pp. 337 ◽  
Author(s):  
H. Lambrechts ◽  
F. E. van Niekerk ◽  
S. W. P. Cloete ◽  
W. A. Coetzer ◽  
G. van der Horst
Keyword(s):  
Adverse Effect ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Sperm Parameters ◽  
Sperm Viability ◽  
Base Population ◽  
Epididymal Sperm ◽  
High Line ◽  
Computer Aided

Microscopically evaluated sperm parameters, as well as computer-aided sperm motility analysis (CASMA), were used to assess sperm quality and the effect of cryopreservation on ram semen obtained from two genetically diverse Merino lines. These lines were divergently selected on maternal ranking values for multiple rearing ability from the same base population since 1986. Replacements in the high (+) line were preferentially the progeny of ewes rearing >1 lamb per joining. Progeny of ewes rearing <1 lamb per joining was preferred as replacements in the low (–) line. Sperm quality, as assessed by percentages of live, abnormal and acrosome-intact spermatozoa as well as by motility, was independent (P≤0.20) of line, time of sampling and their interaction in ejaculated samples obtained from the eight rams used as sires in 1995. Sperm quality of frozen–thawed samples was adversely affected (P≤0.01) by cryopreservation and thawing at 35˚C for 30 s relative to fresh ejaculated samples. No consistent differences between lines were found in epididymal sperm samples obtained from 12 slaughtered rams (6 from each line). The adverse effect (P≤0.05) of cryopreservation and thawing at 35˚C for 30 s on sperm viability and motility was also demonstrated for these samples.

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.

scholarly journals Stallion Sperm Viability, as Measured by the Nucleocounter SP-100, Is Affected by Extender and Enhanced by Single Layer Centrifugation

2010 ◽  
Vol 2010 ◽  
pp. 1-7 ◽  
Author(s):  
J. M. Morrell ◽  
A. Johannisson ◽  
L. Juntilla ◽  
K. Rytty ◽  
L. Bäckgren ◽  
...  
Keyword(s):  
Strong Correlation ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Single Layer ◽  
Sperm Viability ◽  
Flow Cytometric ◽  
New Instrument ◽  
Counting Chamber ◽  
Semen Extender

On-stud assessment of stallion sperm quality can be problematic. A new instrument, the Nucleocounter SP-100, was validated for measuring stallion sperm concentration and viability. It was subsequently used to evaluate sperm viability in Kenney's extender and INRA96. There was a strong correlation between sperm concentrations measured by the Nucleocounter SP-100 and by the Bürker counting chamber (; ). Similarly, there was a good correlation between sperm viability results from the Nucleocounter SP-100 and flow cytometric results (; ). Sperm viability at 24 hours was significantly better for samples extended in INRA96 than in Kenney's extender (). Furthermore, sperm kinematics were better for stored samples in INRA96 than in Kenney's extender. Single Layer Centrifugation selected spermatozoa that maintained their viability better during storage for 24 hours than the uncentrifuged samples. In conclusion, the type of semen extender used and Single Layer Centrifugation were found to influence both the kinematics and viability of stallion spermatozoa. The Nucelocounter-SP100 was considered to be a useful instrument for rapidly measuring stallion sperm concentration and viability.

scholarly journals IMPACT OF AMPK ACTIVATOR METFORMIN ON SPERM QUALITY

2018 ◽  
Vol 15 (4) ◽  
pp. 605-615
Author(s):  
Nguyen Thi Mong Diep ◽  
Elisabeth Blesbois
Keyword(s):  
Sperm Motility ◽  
Acrosome Reaction ◽  
Positive Impact ◽  
Sperm Quality ◽  
Fluorescent Intensity ◽  
Freeze Thaw ◽  
The Media ◽  
Cryopreserved Sperm ◽  
Amp Activated Protein Kinase

Semen cryopreservation allows crucial management of animal genetic diversity. However, the freeze-thaw process causes biochemical and physical alterations, impairing sperm energy-dependent functions. Currently, many chemicals are added to the media to enhance frozen-thawed sperm quality during artificial insemination. The aims of this study was to determine the effects of Metformin (Metf) on fresh chicken sperm motility and ability to perform acrosome reaction, and to evaluate Metf’s effects on the functions of cryopreserved sperm. Chicken semen was diluted and incubated at 35°C in media supplemented with or without different doses of 5’-AMP-Activated Protein Kinase (AMPK) activator, Metf (0,5 to 5 mM). We then looked for the concentration improving the most sperm quality to use it in the cryopreservation media used for chicken sperm. Our results show that 1 mM Metf is the concentration giving the best results regarding sperm quality. AMPKα phosphorylation, viability, acrosome reaction ability (AR), and various motility parameters, were negatively affected by the freeze-thaw process, and that Metf partially restored them. Sperm quality improved (mean increased by 23% for motility, by 10% for viability) as well as AMPKα phosphorylation (mean increased by 30%). Moreover, fluorescent intensity levels of phospho-AMPK were also stronger with Metf than in the control. These results show that the presence of Metf in fresh semen has a positive impact on the quality of sperm and helps reducing the gradual decline in sperm motility caused by cryopreservation by partially restoring several essential sperm functions, and thus leads to a better overall quality of cryopreserved sperm.

scholarly journals Positive effects of trehalose and cysteine on ram sperm parameters

2017 ◽  
Vol 62 (No. 5) ◽  
pp. 245-252 ◽  
Author(s):  
S. Gungor ◽  
C. Ozturk ◽  
AD Omur
Keyword(s):  
Sperm Motility ◽  
Sperm Parameters ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Sperm Viability ◽  
Significant Protection ◽  
Positive Effects ◽  
Liquid Storage ◽  
Acrosome Integrity ◽  
Ram Sperm

The aim of this study was to determine the effects of trehalose and cysteine on sperm motility, viability, mitochondrial activity and acrosome integrity during liquid storage of Merino ram semen. Ejaculates were collected using artificial vaginas from five Merino rams, microscopically evaluated and pooled at 37 °C. The pooled semen samples were diluted in a Tris-based extender, including cysteine (2 mM and 4 mM), trehalose (10 mM and 25 mM) and no antioxidant (control). Diluted semen samples were kept in tubes and cooled from 37 to 5 °C in a cold cabinet, and maintained at 5 °C. Cooled samples were evaluated for sperm motility, viability, mitochondrial activity and acrosome integrity at 0, 24, 48, 72 and 96 h. Extender supplemented with trehalose (10 and 25 mM) and cysteine (2 and 4 mM) led to higher motility in comparison to the control at 24, 48, 72 and 96 h of liquid storage (P &lt; 0.05). Trehalose at the doses of 10 mM, 25 mM and 2 mM cysteine led to higher viability between 24–48–72 h and at 96 h of liquid storage (P &lt; 0.05). Further, 4 mM of cysteine improved sperm viability rates at 24 and 48 h of storage compared to the control group (P &lt; 0.05), and resulted in improved acrosome integrity rates compared to the control group at 72 and 96 h of storage (P &lt; 0.05). Extender supplemented with 10 and 25 mM trehalose at 24 and 72 h and 4 mM cysteine at 24 and 96 h of storage led to higher sperm mitochondrial activity rates when compared to the control group (P &lt; 0.05). In conclusion, the findings of this study show that trehalose and cysteine provided significant protection to ram sperm parameters during liquid storage.

scholarly journals Effects of the platelet-activating factor (PAF) on selected quality parameters of cryopreserved bull semen (AI) with reduced sperm motility

2016 ◽  
Vol 19 (1) ◽  
pp. 147-158 ◽  
Author(s):  
M. Lecewicz ◽  
W. Kordan ◽  
A. Majewska ◽  
S. Kamiński ◽  
A. Dziekońska ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Platelet Activating Factor ◽  
Quality Parameters ◽  
Sperm Viability ◽  
Plasma Membrane Integrity ◽  
Positive Effects ◽  
Bull Semen

Abstract The aim of the study was to determine the effects of platelet-activating factor (PAF) on selected quality parameters of cryopreserved bull semen with reduced sperm motility used for artificial insemination. The aim of experiment 1 was to identify the optimal concentration of the phospholipid able to preserve sperm viability. Cryopreserved semen was treated with different PAF concentrations: 1×10-5M, 1×10-6M, 1×10-7M, 1×10-8M and 1×10-9M. The experiment demonstrated that PAF at concentration 1×10-9M increased most the sperm viability parameters (motility parameters, plasma membrane integrity and mitochondrial function) after 120 min of incubation of thawed semen at 37°C. Cryopreserved bull semen with reduced sperm motility (below 70%) was supplemented with PAF in a concentration of 1×10-9M. A statistically significant increase in sperm motility, percentage of linear motile spermatozoa and VSL value was observed after 120 min incubation of sperm with 1×10-9M PAF. Sperm supplementation with PAF also had positive effects on plasma membrane integrity and percentage of spermatozoa with preserved mitochondrial transmembrane potential, but the differences were not statistically significant. The results indicated positive effects of PAF supplementation at a concentration of 1×10-9M on the selected sperm quality parameters in cryopreserved bull semen with reduced motility.

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