Freezability of Andalusian donkey (Equus asinus) spermatozoa: effect of extenders and permeating cryoprotectants

The aim of this study was to compare the effect of two semen extenders and four permeating cryoprotectants on post-thaw sperm quality of Andalusian donkeys. First, 32 ejaculates were pooled, split and frozen in either Gent B or INRA 96 with egg yolk and glycerol. Second, 12 pooled semen samples were simultaneously frozen in Gent B (glycerol) or Gent A containing ethylene glycol (EG; 1 or 1.5%) or dimethyl sulfoxide (DMSO; 1.5 or 2%). Finally, nine pooled samples were simultaneously cryopreserved in Gent A containing 1% EG (as control), dimethylformamide (DMFA; 1 or 2.5%) or a combination of 1% EG and 1.5% DMFA. Gent B yielded a higher (P < 0.01) post-thaw sperm motility than modified INRA96. EG 1% increased the sperm membrane integrity (P < 0.001), whereas DMSO affected sperm motility and membrane integrity (P < 0.001). DMFA 2.5% yielded higher (P < 0.001) values for sperm motility and membrane integrity. We concluded that Gent B improves in vitro post-thaw sperm quality of donkey spermatozoa, but the replacement of glycerol with 1% EG or 2.5% DMFA increased sperm protection against cryodamage. The use of DMSO for freezing donkey semen was unsuccessful and a toxic effect is suspected. These extenders should be included in the pre-freeze test for each donkey.

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Relationship of extender and packaging system an the length of preservation and the quality of chilled semen of Boer goat

Jurnal Ilmu Ternak dan Veteriner ◽

10.14334/jitv.v21i1.1350 ◽

2016 ◽

Vol 21 (1) ◽

pp. 49

Author(s):

Arie Febretrisiana ◽

. Anwar ◽

Simon Sinulingga

Keyword(s):

Sperm Motility ◽

Sperm Quality ◽

Membrane Integrity ◽

Egg Yolk ◽

Packaging System ◽

Boer Goat ◽

Sperm Membrane ◽

Relationship Of ◽

Artificial Vagina

<p class="abstrak2">The aim of this research was to compare the effectiveness of different extender (either Triladyl or Tris Egg Yolk extender) and different packaging method (pool and straw) of chilled semen an the length of preservation and the quality of chilled semen of Boer goat. Semen was collected using an artificial vagina from 3 two years old Boer bucks with body weight of 50-55 kg. It was evaluated under a microscope, then each was diluted either in Tris egg yolk extender (TEY) or Triladyl. Those diluted sperms were then packed either in pool or straw and preserved at 5⁰C refrigerator. Sperm motility, viability and membrane integrity of each group were evaluated every 24 h for up to 5 days. Results showed that sperm motility in Triladyl of pool packaging system up to 3 days was higher than straw packaging system or TEY in pool or straw packaging system which were 45.8%, 26.1%, 32.1% and 9.1%, respectively (P<0.05). Percentage of sperm membrane integrity showed the same pattern to Triladyl both in pool and straw packaging system which was higher than TEY group (75.2% and 77,2%; P<0.05). Sperm viability in Triladyl both in pool or straw packaging system decreased (P<0.05) after 3 days of preservation (77.1% and 76.2%) but TEY significanly decreased after 4 days of preservation either in pool or straw packaging system (73.2% and 58.0%; P<0.05). It was concluded that sperm quality decreased with increasing of the length of preservation while Triladyl extender in pool packaging system showed the best quality.</p><strong>Key Words: </strong>Chilled Semen, Boer, Triladyl, Tris Egg Yolk, Straw

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Effects of apple and orange juices on quality of refrigerated goat semen

Journal of Agricultural Sciences Belgrade ◽

10.2298/jas1801053a ◽

2018 ◽

Vol 63 (1) ◽

pp. 53-65

Author(s):

Ezekiel Adekunle ◽

James Daramola ◽

Olusiji Sowande ◽

John Abiona ◽

Monsuru Abioja

Keyword(s):

Sperm Motility ◽

Orange Juice ◽

Apple Juice ◽

Membrane Integrity ◽

Egg Yolk ◽

Fruit Juices ◽

Sperm Viability ◽

In Vitro Storage

This study investigated the effects of apple and orange juices on quality of refrigerated spermatozoa of goat bucks. Semen samples from WAD goat bucks were diluted with Tris-egg yolk extenders each supplemented with apple and orange juices at 0, 2.5, 5, 7.5 and 10/100 ml of diluents. The diluted semen samples were assessed for sperm viability and malondialdehyde (MDA) concentration after in vitro storage for 240 hours at 5oC. The ability to maintain sperm motility was higher in the extenders with 7.5% orange juice followed by 10% apple juice compared to other treatments (P<0.05). The extenders supplemented with 2.5%, 5% and 7.5% apple juice, and 5% orange juice had higher intact acrosome compared to other treatments and the control (P<0.05). The 10% orange juice had higher percentage membrane integrity compared to other treatments. Consistent and reduced (P<0.05) MDA levels were observed in the extenders supplemented with fruit juices and lower MDA was observed in the extenders supplemented with 10% apple juice compared to other treatments and the control (P<0.05). The findings reveal that additions of the fruit juices to semen extenders to maintain the viability of refrigerated spermatozoa were best at concentrations of 10 ml/100 ml of apple juice and 7.5 ml/100 ml of orange juice.

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Ram and Goat Semen Immunosexed and Diluted in Powdered Coconut Water-Based Preservation Medium (ACP101/102c).

Acta Scientiae Veterinariae ◽

10.22456/1679-9216.114066 ◽

2021 ◽

Vol 49 ◽

Author(s):

Bruna Farias Brito ◽

Bárbara Mara Bandeira Santos ◽

Leonardo Alves Rodrigues Cabral ◽

Luiz Carlos Pinheiro Maia ◽

Natanael Aguiar Braga Negreiros ◽

...

Keyword(s):

Monoclonal Antibody ◽

Liquid Nitrogen ◽

Sperm Motility ◽

Membrane Integrity ◽

Egg Yolk ◽

Specific Protein ◽

Coconut Water ◽

In Vivo Studies

Background: Sperm sexing aims to separate sperm populations in carriers of the “X” or “Y” chromosome. Currently, flow cytometry is a technique that allows greater accuracy; however, it causes structural changes in sperm, reduces viability, and has a high cost. As a result, other methods have been researched, including immunosexing, which uses monoclonal antibodies to detect sex-specific surface antigens. Thus, the objective of this study was to evaluate the immunosexing technique using a monoclonal antibody against sex-specific protein (HY) in the conservation of ram and goat semen in ACP101/102c.Materials, Methods & Results: Ejaculates from 5 rams and 5 goats were collected with the aid of an artificial vagina; they were evaluated and submitted to the immunosexing protocol, according to the manufacturer's recommendations, using the Monoclonal Antibody Kit specific for mammalian sperm with “Y” chromosomes (HY; HY Biotechnology, Rio de Janeiro, RJ, Brazil). After sexing, the supernatant was resuspended in the cryopreservation diluent: ACP ram (ACP101/102c + 20% egg yolk + 7% glycerol) and ACP goat (ACP101/102c + 2.5% egg yolk + 7% glycerol), packaged in 0.25 mL straws, refrigerated at 4°C, stabilized for 30 min, frozen in liquid nitrogen vapor (-60°C) for 15 min, immersed in liquid nitrogen, and stored in cryogenic cylinders. The samples were evaluated in natura (T1), after immunosexing (T2) and after thawing (T3) for sperm motility subjectively using conventional microscopy (40x). Plasma membrane integrity (IMP) and sperm cell morphology were evaluated by the smear staining technique using eosin-nigrosine dye, and the percentages of healthy and morphologically defect spermatozoa were determined. In the evaluation of ram semen regarding sperm motility and IMP, no statistically significant differences were observed between treatments after sexing in the evaluation of absolute data (P > 0.05), with the difference being observed only between T1 and T2, and T3 (P < 0.05). Regarding the relative percentage and sperm morphology, no statistically significant differences were observed (P > 0.05). Regarding the evaluation of goat semen samples, the motility parameters were consistent with the technique submitted; however, the IMP data did not appear as expected, requiring further evaluation for a better assessment of the technique for this species.Discussion: The data obtained from ram semen submitted to the immunosexing protocol, regarding the absolute evaluation of motility and IMP, demonstrated that the non-sexed semen (T1) was superior to the sexed treatments (T2 and T3); however, it is noteworthy that freezing started with approximately 50% of the cells, since the immunosexing technique results in a loss of viability of approximately 50% of the sperm, which corresponds to the ratio of sperm carrying the X chromosome. In addition, when the data in this study were transformed into relative values, no statistical differences were observed, indicating that the immunosexing protocol, as well as the freezing protocol, did not significantly affect the quality of ram sperm cells. In relation to the immunosexing of goat semen, future studies should be conducted in vitro to define a more appropriate protocol for the species and, in addition, in vivo studies should be performed to prove the quality of the technique. It was concluded that the immunosexing process using a monoclonal antibody against sex-specific protein (HY) associated with the use of powdered coconut water diluent (ACP101/102c) in the cryopreservation of semen proved to be efficient in the in vitro evaluation of ovine species.

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Kriopreservasi Semen Kambing Boer dengan Konsentrasi Pengencer Nira Aren dan Gliserol Berbeda

Jurnal Ilmu dan Teknologi Peternakan Tropis ◽

10.33772/jitro.v6i1.5422 ◽

2019 ◽

Vol 6 (1) ◽

pp. 1

Author(s):

Muhammad Riyadhi ◽

Anis Wahdi ◽

Muhammad Rizal

Keyword(s):

Sperm Motility ◽

Membrane Integrity ◽

Egg Yolk ◽

Boer Goat ◽

Palm Juice ◽

Sperm Membrane ◽

Live Sperm

ABSTRAK Penelitian bertujuan untuk mengetahui efektivitas nira aren sebagai pengencer alternatif dalam proses pembekuan (kriopreservasi) semen kambing boer.Kriopreservasi semen kambing boer menggunakan pengencer tris-gliserol-kuning telur (P1 73-7-20%), nira aren-gliseol-kuning telur(masing-masing P2 74-6-20%, P3 73-7-20%, dan P4 72-8-20%) dan andromed (P5 tanpa mengandung kuning telur dan gliserol). Parameter evaluasi meliputi motilitas, viabilitas, dan membrane plasma utuh setelah pengenceran, ekuilibrasi dan thawing. Evaluasi motilitas pasca thawing menunjukkan P5 52% berbeda nyata (P<0.05) dengan P1 42%, selanjutnya P5 dan P1 berbeda sangat nyata (P<0.05) dengan P2 8%, P3 6% dan P4 12%. Viabilitas pasca thawing menunjukkan P5 65,4% tidak berbeda nyata (P>0,05) dengan P1 61,8%, akan tetapi P5 dan P1 berbeda sangat nyata (P<0.05) dengan P2 26,2%, P3 29,8%, dan P4 34%. Membran plasma utuh (MPU) pasca thawing menunjukkan P5 66,2% tidak berbeda nyata (P>0,05) dengan P1 65,4%, akan tetapi keduanya berbeda sangat nyata (P<0.05) dengan P2 39%, P3 38%, dan P4 36,2%. Disimpulkan kriopreservasi semen kambing boer dengan pengencer nira aren dan gliserol pada konsentrasi berbeda belum dapat dipergunakan sebagai sumber bibit berdasarkan standar nasional Indonesia.Kata Kunci : Kambing boer, semen, nira arenABSTRACTThe experiment was conducted to determine the effect of sugar palm juice as alternative extender for cryopreservation process of boer semen.Tris-glycerol-egg yolk (P1 73-7-20%), Sugar palm juice-glyserol-egg yolk (P2 74-6-20%, P373-7-20%, dan P4 72-8-20%), and andromed (P5) used as a extender in the cryopreservation process of boer semen. Sperm motility (%), live sperm (%) and sperm membrane integrity (%) were recorded after diluted, equilibration and freeze-thawing. Result of post thawing motility showed that P5 52% was significantly different (P <0.05) with P1 42%, then P5 and P1 were significantly different (P <0.05) with P2 8%, P3 6% and P4 12%. Viability after thawing showed P5 65.4% was not significantly different (P> 0.05) with P1 61.8%, but P5 and P1 significantly different (P <0.05) with P2 26.2%, P3 29.8 %, and P4 34%. Spermmembrane integrity post-thawing showed P5 66.2% was not significantly different (P> 0.05) with P1 65.4%, but both were very significantly different (P <0.05) with P2 39%, P3 38% and P4 36.2%. Conclusions, sugar palm juice-glycerol-egg yolk with differentconcentrationsineffectively as an alternative extenderin cryopreservation of boer semen.Keywords: boer goat, semen, sugar palm juice

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Evaluation of sperm quality of Erythrolamprus poecilogyrus sublineatus (Cope, 1860) (Serpentes, Dipsadidae)

Brazilian Journal of Biology ◽

10.1590/1519-6984.19215 ◽

2017 ◽

Vol 77 (3) ◽

pp. 553-557

Author(s):

A. C. Silva ◽

A. S. Varela Junior ◽

T. F. Cardoso ◽

E. F. Silva ◽

D. Loebmann ◽

...

Keyword(s):

Sperm Quality ◽

Membrane Integrity ◽

Coastal Region ◽

Rio Grande ◽

Rio Grande Do Sul ◽

Sperm Analysis ◽

Pampa Region

Abstract Erythrolamprus poecilogyrus sublineatus (Cope, 1860), is a species widely distributed in the Pampa Domain, occurring in Rio Grande do Sul, Argentina and Uruguay, mainlyin the pampa region. In the coastal region of southern Brazil this is serpent is considered one of the most abundant. The purpose of the present study is to describe the techniques of sperm evaluation in vitro for E. poecilogyrus sublineatus in the coastal plain of Rio Grande do Sul, Brazil. After laparatomy the efferent vases were collected and the semen was diluted in 1ml Beltsville Thawing Solution. The characteristics of motility, membrane integrity, mitochondria, acrosome, DNA, cell viability and cellular functionality were evaluated. Fluorescent probes were used for the evaluation of sperm structure in epifluorescence microscope. With the techniques described, it was possible to identify intact and injured cells, enabling the determination of cell characteristics for the spring season (October and November). It was observed in the analyses that 80% of sperm cells were mobile and that 84.1 ± 8.0% of sperm membranes were intact. The standards found were of 48 ± 13.8% of intact acrosome, 73.6 ± 6.0 of perfect DNA and of 91.8 ± 4.0 of functional mitochondria. Thus, these values from the sperm analysis can be used as standards for the species Erythrolamprus poecilogyrus sublineatus.

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PENGGANTIAN BOVINE SERUM ALBUMIN PADA PENGENCER CEP-2 DENGAN SERUM DARAH SAPI DAN PUTIH TELUR TERHADAP KUALITAS SEMEN CAIR SAPI LIMOUSIN SELAMA PENDINGINAN (The Substitution of Bovine Serum Albumin with Cattle Blood Serum and Egg White in CEP-2 Diluent on the Quality of Limousin Bull Liquid Semen during Refrigerating)

Jurnal Kedokteran Hewan - Indonesian Journal of Veterinary Sciences ◽

10.21157/j.ked.hewan.v10i2.5025 ◽

2016 ◽

Vol 10 (2) ◽

pp. 98-102

Author(s):

Trinil Susilawati ◽

Feri Eka Wahyudi ◽

Inna Anggraeni ◽

Nurul Isnaini ◽

Muhammad Nur Ihsan

Keyword(s):

Bovine Serum Albumin ◽

Serum Albumin ◽

Blood Serum ◽

Sperm Motility ◽

Bovine Serum ◽

Sperm Quality ◽

Egg Yolk ◽

Egg White ◽

Cattle Blood

This study aims to determine the effect of the substitution of bovine serum albumin (BSA) with cattle blood serum and egg white in the diluent of Cauda epididymal Plasma 2 (CEP-2) on sperm quality of Limousin cattle during cooling at 3-5 C. The research material used was rejected Limousin bull sperm (motility of 50-60%) from Artificial Insemination Centre Singosari, Malang. This research was a laboratory experiment using a randomized block design which was composed of six treatments with 10 replications, those were T0 as controls ((90% CEP-2 with BSA + 10% egg yolk); T1 (83.84% CEP-2 + 6.16% cattle blood serum + 10% egg yolk); T2 (81.84% CEP-2 + 8.16% cattle blood serum + 10% egg yolk); dan T3 (90% CEP-2 + 0,4% egg white + 10% egg yolk); T4 (90% CEP-2 + 0.8% egg white + 10% egg yolk); and T5 (90% CEP-2 without BSA + 10% egg yolk). Parameters measured were the percentage of motility, viability, and abnormality of sperms. Results of research after 48 hours of storage showed that the percentage of sperm motility in T0, T1, T2, T3, T4, and T5 were 40.50±5.90, 36±36.16, 34.00±6.58, 40.50±3.69, 38.50±3.37, and 38.50±4.12, respectively, while the percentage of sperms viability were 75.16±4.30, 70.50±2.88, 73.80±2.80, 74.80±3.30, 75.13±3.13, and 74.03±4.13, respectively, and the percentage of sperms abnormality were 10.14±2.34, 10.62±1.34, 11.33±2.00, 10.94±2.82, 10.02±1.95, and 10.78±1.96, respectively. In conclusion, CEP-2 diluent with or without the addition of 19% egg yolk in BSA and the substitution of BSA with 0.4-0.8% egg white can maintain semen quality to hour of 48 in cold storage.This study aims to determine the effect of the substitution of bovine serum albumin (BSA) with cattle blood serum and egg white in the diluent of Cauda epididymal Plasma 2 (CEP-2) on sperm quality of Limousin cattle during cooling at 3-5 C. The research material used was rejected Limousin bull sperm (motility of 50-60%) from Artificial Insemination Centre Singosari, Malang. This research was a laboratory experiment using a randomized block design which was composed of six treatments with 10 replications, those were T0 as controls ((90% CEP-2 with BSA + 10% egg yolk); T1 (83.84% CEP-2 + 6.16% cattle blood serum + 10% egg yolk); T2 (81.84% CEP-2 + 8.16% cattle blood serum + 10% egg yolk); dan T3 (90% CEP-2 + 0,4% egg white + 10% egg yolk); T4 (90% CEP-2 + 0.8% egg white + 10% egg yolk); and T5 (90% CEP-2 without BSA + 10% egg yolk). Parameters measured were the percentage of motility, viability, and abnormality of sperms. Results of research after 48 hours of storage showed that the percentage of sperm motility in T0, T1, T2, T3, T4, and T5 were 40.50±5.90, 36±36.16, 34.00±6.58, 40.50±3.69, 38.50±3.37, and 38.50±4.12, respectively, while the percentage of sperms viability were 75.16±4.30, 70.50±2.88, 73.80±2.80, 74.80±3.30, 75.13±3.13, and 74.03±4.13, respectively, and the percentage of sperms abnormality were 10.14±2.34, 10.62±1.34, 11.33±2.00, 10.94±2.82, 10.02±1.95, and 10.78±1.96, respectively. In conclusion, CEP-2 diluent with or without the addition of 19% egg yolk in BSA and the substitution of BSA with 0.4-0.8% egg white can maintain semen quality to hour of 48 in cold storage.

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Efficiency of Commercial Egg Yolk-Free and Egg Yolk-Supplemented Tris-Based Extenders for Dromedary Camel Semen Cryopreservation

Animals ◽

10.3390/ani9110999 ◽

2019 ◽

Vol 9 (11) ◽

pp. 999 ◽

Cited By ~ 5

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.

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180 ASSESSMENT OF BULL SEMEN QUALITY LOADED IN NEW SensiTemp STRAWS USING SEMEN AND IN VITRO PRODUCTION TECHNOLOGIES

Reproduction Fertility and Development ◽

10.1071/rdv28n2ab180 ◽

2016 ◽

Vol 28 (2) ◽

pp. 221

Author(s):

D. Le Bourhis ◽

S. Camugli ◽

P. Salvetti ◽

L. Schibler ◽

E. Schmitt

Keyword(s):

Sperm Motility ◽

Semen Quality ◽

Sperm Quality ◽

Semen Analysis ◽

Membrane Integrity ◽

Computer Assisted ◽

Cleavage Rate ◽

Fluid Medium ◽

And Control

SensiTemp, a new in vitro maturation (IMV) bull straw concept, presents the advantage of colour changing while the straw is thawed. The colour of frozen straws is blue and straws start to become white when the temperature reaches 33°C, with a complete change of colour at 37°C. The objective of this study is to assess sperm quality after thawing of semen frozen in SensiTemp from 2 bulls, by analysing, in experiment 1, sperm motility and membrane integrity using computer-assisted semen analysis (CASA) and flow cytometry (FC), and, in experiment 2, the in vitro embryo production (IVP) using IVP technologies [IVM, IVF, and in vitro culture (IVC)]. The ejaculates of 2 bulls, selected during preliminary experiments on high in vitro fertility, were harvested at CIA L’Aigle, France, and split ejaculates were frozen in experimental (SensiTemp) and conventional (control) straws. In experiment 1 after thawing semen from the 2 types of straws (5 pooled straws each; 2 replicates), motility was assessed using the IVOS CASA system (Hamilton Thorne Inc., Beverly, MA, USA) and membrane integrity was evaluated through FC with Cytosoft software (Millipore-Guava Technologies Inc., Hayward, CA, USA). In experiment 2, IVF was used to evaluate the non-toxicity of SensiTemp and control straws. Cumulus-oocyte complexes (COC; n = 1178; 4 replicates) collected from slaughterhouse ovaries were matured in IVM medium (TCM-199 with bicarbonate, Sigma-Aldrich, Saint Quentin Fallavier, France; 10 µg mL–1 FSH-LH, Reprobiol, Liège, Belgium; and 10% FCS, Thermo Fisher, Illkirch, France) for 22 h. After fertilization, presumptive zygotes of each group (SensiTemp and control for each bull) were cultured in synthetic oviduct fluid medium (SOF, Minitube, Tiefenbach, Germany) with 1% estrous cow serum (ECS) and 0.6% BSA (Sigma-Aldrich, France) up to 8 days. All cultures were conducted at 38.5C in 5% CO2, and 5% O2. The cleavage and blastocysts rates were evaluated on Days 3 and 7, respectively, for each group. Embryo quality was recorded on Day 7 according to the IETS evaluation. Data from each bull were analysed separately using the chi-squared test (P < 0.05). In experiment 1, neither sperm motility from bull 1 (61.2 and 60.5%) and bull 2 (66.2 and 66.5%) nor membrane integrity from bull 1 (58.6 and 52.2%) and bull 2 (61.0 and 61.9%) were different between SensiTemp and control, respectively. Results from experiment 2 showed no difference (P > 0.05) in cleavage rate between SensiTemp and control for the 2 bulls: 92.1 and 91.7% for bull 1 and 94.2 and 94.6% for bull 2 respectively. The blastocysts rate on Day 7 did not differ (P > 0.05) among groups (47.5, 47.1 and 51.3, 50.4% for SensiTemp and control bull 1 and bull 2, respectively) nor the quality of embryos retrieved in the different groups: 25.4, 23.3, and 30.8, 29.6% in grade 1 embryo for SensiTemp and control bull 1 and bull 2, respectively. Those results demonstrate, in vitro, that the new SensiTemp straws were non-toxic and did not affect the semen quality after thawing nor did the SensiTemp straws affect the ability of sperm cells to fertilize oocytes and produce 8-day-old embryos.

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Effect of Glutathione on Sperm Quality in Guanzhong Dairy Goat Sperm During Cryopreservation

Frontiers in Veterinary Science ◽

10.3389/fvets.2021.771440 ◽

2021 ◽

Vol 8 ◽

Author(s):

Jiahao Zou ◽

Lixuan Wei ◽

Dexian Li ◽

Yongtao Zhang ◽

Guang Wang ◽

...

Keyword(s):

Plasma Membrane ◽

Sperm Motility ◽

Sperm Quality ◽

Membrane Integrity ◽

Total Power ◽

Dairy Goat ◽

Control Group ◽

Membrane Function ◽

Motility Parameters

In the process of cryopreservation of dairy goat semen, it will face many threats such as oxidative damage, which will affect the motility and plasma membrane function of sperm. As an endogenous antioxidant in animals, glutathione (GSH) can significantly improve the quality of thawed sperm when added to the frozen diluent of semen of pigs and cattle. In this study, different concentration gradients of GSH [0 mmol/L (control), 1, 2, 3, 4 mmol/L] were added to the frozen diluent of Guanzhong dairy goat semen. By detecting the sperm motility parameters, acrosome intact rate and plasma membrane intact rate after thawing, the effect of GSH on the cryopreservation of dairy goat semen was explored. Sperm motility parameters were measured with the computer-aided sperm analysis (CASA) system (total power, TM; forward power, PM; linearity, LIN; average path speed, VAP; straight line speed, VSL; curve speed, VCL; beat cross frequency, BCF). The sperm acrosome integrity rate after thawing was detected by a specific fluorescent probe (isothiocyanate-labeled peanut agglutinin, FITC-PNA), and the sperm plasma membrane integrity rate after thawing was detected by the hypotonic sperm swelling (HOST) method. Reactive oxygen species (ROS) kit, malondialdehyde (MDA) kit, superoxide dismutase (SOD) kit, glutathione peroxidase (GSH-PX) kit were used to detect various antioxidant indicators of thawed sperm. in vitro fertilization experiment was used to verify the effect of adding glutathione on sperm fertilization and embryo development. The results showed that when the concentration of glutathione was 2 mmol/l, the sperm viability, plasma membrane intact rate, and acrosome intact rate were the highest after thawing, reaching 62.14, 37.62, and 70.87% respectively, and they were all significantly higher. In terms of antioxidant indexes; the values of SOD and GSH-PX were 212.60 U/ml and 125.04 U/L, respectively, which were significantly higher than those of the control group; The values of ROS and MDA were 363.05 U/ml and 7.02 nmol/L, respectively, which were significantly lower than the control group. The addition of 2 mmol/L glutathione significantly improves the fertilization ability of sperm. In short, adding 2 mmol/l glutathione to the semen diluent can improve the quality of frozen Guanzhong dairy goat sperm.

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Efektivitas Penambahan Vitamin E (alfa-Tokoferol) dalam Medium Pencucian Sperma dengan Sentrifugasi terhadap Kualitas Spermatozoa Sapi Brahman

Jurnal Agripet ◽

10.17969/agripet.v12i2.196 ◽

2012 ◽

Vol 12 (2) ◽

pp. 7-13

Author(s):

Dasrul Dasrul ◽

Rasmaidar Rasmaidar ◽

Abdul Harris

Keyword(s):

Vitamin E ◽

Sperm Quality ◽

Membrane Integrity ◽

Average Percentage ◽

Treatment Groups ◽

Frozen Semen ◽

Brahman Cattle ◽

Sperm Membrane ◽

Better Than

Effect of vitamin E addition (alfa-tokoferol) into sperm washing medium by centrifuge on the quality of Brahman cattle spermatozoa ABSTRACT. The aims of study to determine the effectiveness of the addition of vitamin E in the washing medium by centrifugation on sperm quality Brahman cattle. frozen semen of Brahman cattle, divided into 4 treatment groups addition of vitamin E in the washing medium: 0.0gr/100 ml medium (K0), 0.1gr /100 ml medium (K1); 0.2gr/100 ml medium (K2) and 0.3 g / 100 ml medium (K4), each group was repeated 5 times. Examination of motility, viability and integrity of sperm membrane performed according to WHO standards. The data obtained were analyzed with one-way ANOVA and Duncan test. The average percentage of motility, viability and membrane integrity of spermatozoa in the addition of vitamin E were significantly different (P 0.05) compared to the control. Percentage of motility, viability and membrane integrity of spermatozoa in the group K2 significantly higher (P 0.05) compared with the group K3: K1 and K0. Percentage of motility, viability and sperm capacitation and sperm live on group K3 significantly higher (P 0.05) compared with the K1 and K0. While the percentage motility of spermatozoa in the group K1 higher were not significant (P 0.05) compared with the group K0. The addition of vitamin E in the medium on the process of washing spermatozoa Brahman cattle. The addition of vitamin E 0.2gr/100ml better than vitamin E 0.1gr/100ml and 0.3gr/100ml in maintaining the percentage of motility and live spermatozoa Brahman cattle.

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