scholarly journals L-Carnitine Improves Cytoprotection during Cryopreservation: A case study on Nili-Ravi Buffalo Sperm

2021 ◽  
pp. 80-85
Author(s):  
Saima Qadeer ◽  
Rabea Ejaz ◽  
Asma Ul Husna ◽  
Asima Azam ◽  
Syeda Laila Rubab ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Chromatin ◽  
Antioxidative Effect ◽  
Sperm Plasma Membrane ◽  
Cryopreserved Sperm ◽  
Chromatin Integrity ◽  
Live Sperm ◽  
Artificial Vagina

The current study was aimed to evaluate the antioxidative effect of L-Carnitine at post thawing following cryopreservation of Nili-Ravi buffalo sperm. For this purpose, semen from three buffalo bulls were collected for 3 weeks using an artificial vagina (N=18; replicates). The qualified ejaculates were diluted employing tris-citric acid extender i.e., control did not receive any L-Carnitine and experimental groups having 0.5, 1.0, and 1.5 ng/mL of L-carnitine at 37 C with approximately 50 x 106 sperm/mL. The semen was cooled at 4 C and then equilibrated (4 hours), filled in straws (0.5 mL) at4 C, placed on LN2 vapours for 10 min, and kept into an LN2 container. The thawed semen was evaluated for post-thaw quality. The integrity of the sperm plasma membrane and motility (P?0.05) was highest in the extenders having 1.0 ng/mL of L-carnitine as compared to the control(received no L-Carnitine). However, sperm chromatin integrity and viability(live sperm with intact acrosome) remained similar. It was concluded that supplementing 1.0 ng/mL L-Carnitine of extender can improve the post-thaw quality of cryopreserved sperm. Based on the results of the current experiments it is recommended to include L-carnitine extender to improve post-thaw quality of buffalo sperm in terms of its motility and integrity ofits plasma membrane. Keywords: Buffalo, Sperm, Cryopreservation, Extender, L-Carnitine, Artificial insemination.

In vitro Supplementation of Linoleic Acid Improves Quality of Cryopreserved Buffalo Semen

2020 ◽  
Author(s):  
R Ejaz ◽  
S Qadeer ◽  
M S Ansari ◽  
B A Rakha ◽  
S Shamas ◽  
...  
Keyword(s):  
Linoleic Acid ◽  
Membrane Integrity ◽  
Sperm Chromatin ◽  
Standard Procedures ◽  
Buffalo Semen ◽  
Chromatin Integrity ◽  
Live Sperm ◽  
And Control

Present study was designed to evaluate the effect of linoleic acid (LA) supplementation in extender on post thaw quality of cryopreserved buffalo semen. Semen was collected from three adult Nili Ravi buffalo bulls of same age with artificial vagina (42°C) for five weeks (replicates; N=30). Qualified semen ejaculates (>1mL volume, >60% motility, >0.5 billion/mL concentration) were diluted in tris-citric acid extender containing 0.0 (control), 5.0, 10.0 and 20.0ng mL-1 of LA and were cryopreserved using standard procedures. Sperm motility and plasma membrane integrity were improved plessthan0.05) in extender containing 10.0 ng mL-1 of LA compared to other treatments and control while number of acrosome intact live sperm, chromatin integrity and number of morphologically normal sperms remained the same. In conclusion, LA supplementation in extender at 10.0 ng mL-1 was found to be beneficial to improve post thaw quality of cryopreserved buffalo semen.

Interactions between gametes leading to fertilization: the sperm's eye view

1995 ◽  
Vol 7 (4) ◽  
pp. 927 ◽  
Author(s):  
BT Storey
Keyword(s):  
Plasma Membrane ◽  
Acrosome Reaction ◽  
Mammalian Species ◽  
Sperm Chromatin ◽  
Peptide Backbone ◽  
Mouse Sperm ◽  
Gamete Interactions ◽  
Sperm Plasma Membrane ◽  
Sperm Penetration ◽  
Strong Candidate

Sexual reproduction requires that the gamete carrying the male-derived haploid chromatin join with the gamete carrying the female-derived haploid chromatin during fertilization to produce the diploid zygote. To accomplish this feat, the sperm must not only meet the egg, it must recognize the egg and be recognized in turn by the egg, and in the end must enter and be engulfed by the egg. In this selective overview of gamete interactions that lead to fertilization, encounters of three kinds, followed by the finale of gamete fusion, are considered from the sperm's viewpoint, with particular emphasis on the mammalian species with the mouse as the principal model. The first encounter is with the zona pellucida of the egg, to whose surface the sperm must bind. Mouse sperm appear to have four binding sites for zona ligands. Three interact with sugar moieties of the oligosaccharide chains of the mouse zona glycoprotein ZP3; the fourth binds a peptide backbone arginine. Capacitation is not required for this encounter, but is obligate for the second encounter--induction of the acrosome reaction in the bound sperm. The acrosome reaction is an exocytotic process that makes available the enzymatic machinery needed for sperm penetration the zona which is the end point of a sequence of reactions directed by intracellular signalling systems. In mouse sperm, these systems are presumed to be activated by ligands on ZP3 binding to ligand-specific sperm receptors with consequent aggregation of receptors. No receptor has been identified with certainty, nor have candidates for putative ZP3 ligands been identified. Completion of the acrosome reaction allows the sperm to penetrate the zona and, bind to the egg plasma membrane, thereby completing the third encounter. In the mouse, a 94-kDa protein appears essential for this binding. In the guinea-pig, a sperm plasma membrane protein (formerly PH-30, now fertilin), is a strong candidate for the mediator of the fusion process by which the egg engulfs the sperm. Decondensation of the sperm chromatin reverses the remarkable packing of DNA organized by sperm protamines. Mitochondrial DNA is also engulfed by the egg; the question of whether this DNA makes a small finite, or null, contribution to cytosolic inheritance is still in debate. The puzzles attending these encounters are presented as reminders of the intricacy and fascination, as well as of the vital necessity, of gamete interaction.

scholarly journals Effects of the addition of docosahexaenoic acid and ?-tocopherol on quality of equine spermatozoa stored at 5°C

2020 ◽  
Vol 41 (1) ◽  
pp. 167 ◽  
Author(s):  
Breno Fernandes Barreto Sampaio ◽  
Bruno Gomes Nogueira ◽  
Maria Inês Lenz Souza ◽  
Eliane Vianna da Costa-e-Silva ◽  
Carmem Estefânia Serra Neto Zúccari
Keyword(s):  
Lipid Peroxidation ◽  
Plasma Membrane ◽  
Docosahexaenoic Acid ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Membrane Composition ◽  
Artificial Vagina

Plasma membrane composition has impact on phase transition from liquid crystal to gel state of cooled sperm cell. The incorporation of polyunsaturated fatty acids increases its fluidity and can contribute to sperm motility. The aim of this study was to compare the effect of adding docosahexaenoic acid (DHA) and ?-tocopherol (?-Toh) to the cooling extender, singly or combined, to the equine sperm parameters, submitted to cooling, up to 72 hours. Two ejaculates of ten stallions collected with artificial vagina were used, and evaluated for motility, plasma membrane integrity, chromatin fragmentation, mitochondrial activity and lipid peroxidation, according to the following treatments: C; DHA; ?-Toh; DHA/?-Toh; EtOH 100: and EtOH 140 (corresponding to control; 10 ng mL-1 of DHA; 2 mM of ?-Toh; : 10 ng mL-1 of DHA + 2 mM of ?-Toh; 100 µL of ethanol and 140 µL of ethanol respectively). DHA treatment showed higher motility (68.2 ± 12.3; p < 0.05) when compared to control (62.1 ± 16.2), DHA/?-Toh (61.3 ± 12.7) and EtOH (58.1 ± 8.6) groups. In lipid peroxidation assay, the control group showed 2,506.2 ± 796.4 ng of MDA 108 spermatozoa-1, being significantly higher (p < 0.05) than the groups treated with DHA (2,036.0 ± 687.0), ?-Toh (1,890.8 ± 749.5) and DHA/?-Toh (1,821.1 ± 627.2). In conclusion, ?-Toh was effective in diminishing lipid peroxidation of equine sperm subjected to cooling, and DHA improved sperm motility and, in spite of being a polyunsaturated fatty acid with high susceptibility to peroxidation, reduced lipid peroxidation.

scholarly journals Semen Quality of Garut Rams feed by Different Protein Sources and Their Implementation Potential in Small Farms of West Java

2020 ◽  
Vol 20 (1) ◽  
pp. 47-55
Author(s):  
Kurnia Bagus Ariyanto ◽  
Lilis Khotijah ◽  
Dewi Apri Astuti ◽  
Raden Iis Arifiantini ◽  
Jean-Baptise Menassol
Keyword(s):  
Plasma Membrane ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Black Soldier Fly ◽  
Hermetia Illucens ◽  
Brachiaria Humidicola ◽  
Plasma Membrane Integrity ◽  
Protein Sources ◽  
Sperm Plasma Membrane

ABSTRACT. Maggot Hermetia illucens (Maggot Black Soldier Fly, MBSF) is an alternative protein source besides soybean meal (SBM) which may be used as a feed for improving the quality of semen particularly in Garut rams to support prolific nature. The aims of this study were to analyzed and compare the impact of different protein sources in feed on semen quality of Garut rams, and to assess the prediction ability of Garut rams to serve ewe in small-scale breeders in West Java, Indonesia. This study was conducted using a completely randomized design with 3 treatments and 4 replications, consisted of Brachiaria humidicola (BH) grass and T1 (concentrate contains 20% of SBM), T2 (concentrate contains 10% of SBM and 10% of MBSF), and T3 (concentrate contains 20% of MBSF). The parameters measured were feed consumption, semen quality (macroscopic and microscopic characteristics), also a potential ability of rams to serve ewe. The results showed there were no significant effect on protein consumption, semen volume, semen pH, semen color and consistency, sperm mass movement, sperm motility, sperm concentration, sperm morphology, and prediction potential ability to serve ewe. However, the result showed a significant effect (P0.05) on sperm viability and sperm plasma membrane integrity. Sperm plasma membrane integrity of ram feed with T3 was better than T1 and T2 (P0.05). The prediction potential ability rams to serve ewes on MBSF treatment was 38 heads, while in T1 and T2 were 43 and 57 heads, respectively. In conclusion, MBSF can be an alternative source of protein besides SBM to improve the semen quality of Garut rams.  ABSTRAK. Maggot Hermetia illucens (Maggot Black Soldier Fly; MBSF) adalah sumber protein alternatif selain bungkil kedelai (SBM) yang dapat dipergunakan sebagai pakan untuk memperbaiki kualitas semen terutama pada domba Garut untuk mendukung sifat prolifik. Tujuan penelitian ini adalah menganalisis dan membandingkan dampak pemberian sumber protein berbeda terhadap kualitas semen domba Garut dan untuk menilai kemampuan domba Garut pejantan dalam melayani betina pada peternakan rakyat di Jawa Barat, Indonesia. Penelitian ini menggunakan Rancangan Acak Lengkap (RAL) dengan 3 perlakuan dan 4 ulangan yang terdiri dari rumput Brachiaria humidicola (BH) dan T1 (konsentrat mengandung 20% SBM), T2 (konsentrat mengandung 10% SBM dan 10% MBSF), dan T3 (konsentrat mengandung 20% MBSF). Parameter yang diukur adalah konsumsi pakan, karakteristik semen (makroskopis dan mikroskopis) serta potensi domba jantan melayani betina. Hasil Penelitian menunjukkan tidak ada perbedaan signifikan pada konsumsi protein pakan, volume semen, pH semen, warna dan konsistensi semen, gerakan massa sperma, motilitas sperma, konsentrasi sperma, morfologi sperma, dan prediksi potensi pejantan dalam melayani betina. Namun, hasil penelitian menunjukkan terdapat perbedaan (P0.05) pada viabilitas sperma dan membran plasma utuh sperma. Membran plasma utuh pada perlakuan T3 lebih baik dibandingkan perlakuan T1 dan T2 (P0.05). Prediksi potensi betina terlayani dari pejantan yang diberi pakan MBSF adalah 38 ekor, sedangkan yang diberi SBM dan kombinasinya adalah 43 dan 57 ekor. Kesimpulan penelitian ini adalah MBSF dapat menjadi alternatif sumber protein selain bungkil kedelai dalam memperbaiki kualitas sperma domba Garut.

22 EFFECT OF CHOLESTEROL ADDITION TO EQUINE SPERM MEMBRANE ON FERTILITY

2013 ◽  
Vol 25 (1) ◽  
pp. 158
Author(s):  
F. P. Hartwig ◽  
F. P. Lisboa ◽  
G. A. Monteiro ◽  
R. R. D. Maziero ◽  
M. A. Alvarenga ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Quality ◽  
Skim Milk ◽  
Fisher Exact Test ◽  
Exact Test ◽  
Horse Breeding ◽  
Sperm Plasma Membrane ◽  
Sperm Membrane

Artificial insemination with cooled-shipped semen has been widely used in horse breeding. However, some stallions, referred to as poor coolers, present abrupt fertility decrease when their semen is processed, cooled, and transported. Cholesterol incorporation into sperm membranes improves the quality of cryopreserved semen by increasing the sperm membrane stability and fluidity at low temperatures. Despite the beneficial effect of cholesterol addition on sperm quality, studies demonstrate that the presence of large amounts of cholesterol in the plasma membrane interferes with the physiological process of sperm capacitation and is detrimental to frozen equine sperm fertility. The aim of this study was to assess the fertility of cooled semen from good-cooler and poor-cooler stallions after adding cholesterol to sperm membranes. Two stallions were used and classified as good cooler (n = 1) and poor cooler (n = 1) based on the ability to maintain sperm progressive motility after 24 h of cooling at 5°C. For classification of the stallions, the fertility history was also taken into account through the results of pregnancy per cycle using inseminations with cooled semen (<50% for poor cooler and >70% for good cooler). Ejaculates of these stallions were subjected to 2 treatments: control (CON) and cholesterol (CLC). In the CON group, the semen was extended to 30 × 106 sperms mL–1 with skim milk-based extender (BotuSemen™). In the CLC group, the semen was extended as in the CON group plus 0.25 µL/1 × 106 sperms of 6.1 mM cholesterol-loaded cyclodextrin was added. Afterwards, both treatments were cooled at 5°C for 24 h. To test the fertility of poor-cooler and good-cooler stallions, 2 cycles from 25 mares and 2 cycles from 10 mares were respectively used. For both stallions, randomly for each mare, the inseminations were performed by alternating both treatments. If the mare was first inseminated with the CLC treatment, in the next cycle the CON treatment was used and vice versa. After 24 h of ovulation induction, the inseminations were done in the uterine body with 1 × 109 viable cells. Statistical analyses were performed using the Fisher exact test and significance was set at P < 0.05. For the poor cooler, the CON treatment presented 44% pregnancy/cycle compared to 76% for the CLC treatment (11/25a v. 19/25b). For the good cooler, both treatments presented 80% (8/10) pregnancy/cycle. The results suggest that the fertility capability of stallions is not affected by incorporation of cholesterol-loaded cyclodextrin to the sperm plasma membrane. Additionally, the utilization of cholesterol-loaded cyclodextrin may be an option to enable the utilization of cooled-shipped semen from poor cooler stallions for AI programs.

scholarly journals Effect of Monthly Variations on the Plasma Membrane, Acrosome and DNA Integrities of Spermatozoa in Friesian Holstein Bulls Born in Iraq

2020 ◽  
Vol 13 (2) ◽  
pp. 118-124
Author(s):  
Qusay Aboud ◽  
Saad Hatif
Keyword(s):  
Plasma Membrane ◽  
Artificial Insemination ◽  
Significant Decline ◽  
Dna Integrity ◽  
Frozen Semen ◽  
Negative Effect ◽  
Holstein Bulls ◽  
Monthly Variations ◽  
Artificial Vagina

This study was aimed to evaluate the influence of months of the year on the quality of semen in Holstein bulls. A study carried out at artificial insemination centre/ Abou-Ghareeb/ western of Baghdad. A total of 160 ejaculates were collected from 15 bulls born in Iraq via the artificial vagina. The age of the bulls ranged between 4 to 5 years and the study period were December to March and September. The semen samples were diluted with Tris base extender. The semen was packed in a straw according to the program of artificial insemination centre. Semen characteristics (plasma membrane, acrosome, and DNA integrities) were evaluated. The results revealed a significant decrease (P≤0.01) in the plasma membrane and acrosome integrity in September as compared with December, January, February, and March. There was a significant decline (P≤0.05) in DNA integrity in September as compared with December, January, February, and March in fresh and frozen semen. In conclusion, the September month had a negative effect on the plasma membrane, acrosome, and DNA percentage in all bulls.

scholarly journals The Quality of Boer Goat Semen Preserved with Sugar Palm Juice

2018 ◽  
Vol 42 (2) ◽  
Author(s):  
Muhammad Rizal ◽  
Muhammad Riyadhi ◽  
Abrani Sulaiman
Keyword(s):  
Plasma Membrane ◽  
Seminal Plasma ◽  
Egg Yolk ◽  
Boer Goat ◽  
The Third ◽  
Palm Juice ◽  
Boer Goats ◽  
Fresh Semen ◽  
Artificial Vagina

The objective of this study was to examine the effect of seminal plasma on viability of Boer goat spermatozoa and effectiveness of sugar palm juice as an alternative extender during preservation at 5ºC. Semen of two Boer goats were collected using an artificial vagina. Fresh semen were evaluated and divided in equal volume into four tubes. Semen in the first and second tubes diluted with 80% sugar palm juice + 20% egg yolk (P1) and Andromed (P2), respectively. Semen in the third and fourth tubes were centrifuged with 3,000 RPM for 20 minutes, and the supernatant removed. diluted with 80% sugar palm juice + 20% egg yolk (P3) and Andromed (P4), respectively. Diluted-semen were preserved in refrigerator at 5oC, and quality of the spermatozoa including motile spermatozoa (MS), live spermatozoa (LS), and intact plasma membrane (IPM) were evaluated every day for four days. Results of this study showed that at day-2 preservation, mean percentages of MS, LS, and IPM for P2 (72, 83.4, and 83.4%), P3 (72, 82.6, and 82.2%), and P4 (72, 83, 83.8%) were significantly (P<0.05) higher than P1 (3, 24.8, and 25.2%). At day-3 preservation, mean percentages of MS, LS, and IPM for P2 (57, 65.6, and 69.6%) was significantly (P<0.05) higher than P3 (21, 34.8, and 31.8%), P4 (22, 33.6, and 31.2%), and P1 (0, 0, and 0%). In conclusion, semen of Boer goat to be preserved with extender containing egg yolk should be removed seminal plasma. Sugar palm juice containing egg yolk could be used as an extender for Boer goat semen, but should be applied in the AI program immediately after the semen is diluted.

Extender supplementation with catalase maintains the integrity of sperm plasma membrane after freezing–thawing of semen from capuchin monkey

Zygote ◽  
2017 ◽  
Vol 25 (2) ◽  
pp. 231-234 ◽  
Author(s):  
Danuza L. Leão ◽  
Adriel B. Brito ◽  
Stefânia A. Miranda ◽  
Karol G. Oliveira ◽  
Débora V.C. Almeida ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Capuchin Monkey ◽  
Plasma Membrane Integrity ◽  
Sapajus Apella ◽  
Sperm Plasma Membrane ◽  
Cryopreserved Sperm

SummaryWe aimed to evaluate the effect of supplementation of ACP-118® extender with the antioxidant catalase (10 and 50 µg/ml) on Sapajus apella sperm motility, vigour, and plasma membrane integrity during the processes of seminal liquefaction, cooling, and freezing. Catalase did not affect any of the evaluated parameters after semen dilution or cooling. Cryopreserved sperm in the presence of 50 µg/ml catalase presented a plasma membrane integrity similar to that fresh sperm, however.

scholarly journals Freeze-thawing impairs the motility, plasma membrane integrity and mitochondria function of boar spermatozoa through generating excessive ROS

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Bin Zhang ◽  
Yan Wang ◽  
Caihong Wu ◽  
Shulei Qiu ◽  
Xiaolan Chen ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Chromatin Structure ◽  
Membrane Integrity ◽  
Sperm Chromatin ◽  
Mitochondrial Activity ◽  
Atp Content ◽  
Plasma Membrane Integrity ◽  
Freeze Thaw ◽  
Boar Spermatozoa

Abstract Background Cryopreservation is an efficient way to store spermatozoa and is closely associated with the quality of sperm after the freeze-thaw process. During freeze-thaw cycling, excessive reactive oxygen species (ROS) are produced, and the effects of ROS on boar sperm during cryopreservation have not been identified. Results In this study, we evaluated the quality of boar spermatozoa in different steps of cryopreservation (extension, cooling, and thawing for 30 min and 240 min) with or without boar-sperm antioxidant (N-acetylcysteine (NAC)). The ROS levels, sperm motility, plasma membrane integrity, mitochondrial activity, sperm chromatin structure, ATP content, and sperm apoptosis were assayed. After thawing, the ROS level and sperm apoptosis were significantly increased, and the sperm motility, plasma membrane integrity, mitochondrial activity, sperm chromatin structure, and ATP content were significantly impaired compared with those at the extension period and cooling period. Moreover, the addition of N-acetyl L-cysteine (NAC) reversed these changes. Conclusion The freeze-thawing of boar spermatozoa impaired their motility, plasma membrane, mitochondrial activity, sperm chromatin structure and apoptosis by producing excessive ROS. Thus, the downregulation of ROS level by antioxidants, especially the NAC, is important for manufacturing frozen pig sperm to increase reproductive cells and livestock propagation, as well as to improve the application of frozen semen in pigs worldwide.

scholarly journals PENAMBAHAN L – ARGININ DALAM PENGENCER TRIS KUNING TELUR SETELAH EKUILIBRASI 2 JAM TERHADAP MOTILITAS DAN MEMBRAN PLASMA UTUH SPERMATOZOA DOMBA SAPUDI

2020 ◽  
Vol 7 (2) ◽  
pp. 86
Author(s):  
Hastini Suryaningsih ◽  
Emy Koestanty Sabdoningrum ◽  
Suherni Susilowati ◽  
Trilas Sardjito ◽  
Wurlina Wurlina ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Analysis Of Variance ◽  
Sperm Motility ◽  
Egg Yolk ◽  
Equilibration Time ◽  
Sperm Plasma Membrane ◽  
Artificial Vagina ◽  
Plasm Membrane

This research was aimed to determine the addition of L-arginine on motility and sperm plasma membrane of Sapudi sheep after two hour equilibration time in egg yolk tris diluent. This research used fresh samples of sheep Sapudi semen collected by artificial vagina, then were devided into 4 treatments and 6 replications. Analysis of the data using Analysis of Variance (ANOVA) then proceed to the Duncan Test to determine significant differences between treatments. The first treatment P0 was no L – Arginine added as control. P1 was treated with 0,004 M L – Arginine. P2 was treated with 0,005 M L – Arginine, P3 was treated with 0,006 M L – Arginine. Result of two hour equilibration time for the sperm motility showed: (P0) 46,67a   5,16, (P1) 49,16ab   5,84, (P2) 55,00bc   7,74, (P3) 58,33c   6,83. Result of two hour equilibration time for the sperm plasm membrane were: (P0) 39,33a   4,92, (P1) 43,33ab   4,22, (P2) 46,50 bc   3,44, (P3) 50,83 c   3,48. The addition of L-arginine with a concentration as much as 0.006 M, shows the highest result in sperm motility and sperm plasm membrane intact of Sapudi semen.

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