Effects of adding taurine and trehalose to a tris-based egg yolk extender on buffalo (Bubalus bubalis) sperm quality following cryopreservation

SummaryThe aim of this study was to establish a functional freezing–thawing protocol for epididymal sperm of collared peccaries (Pecari tajacu L., 1758) by comparing different extenders. The epididymal sperm from 12 sexually mature males was recovered by retrograde flushing using Tris-based or coconut water-based (ACP®-116c) extenders. After initial evaluation, samples were diluted and frozen with the same extenders to which 20% egg yolk and 6% glycerol were added. After 2 weeks, thawing was performed at 37°C/60 s and sperm motility, vigour, morphology, functional membrane integrity, sperm viability, sperm plasma membrane integrity, and a computer-assisted semen analysis (CASA) were assessed. In addition, to evaluate the survival of frozen–thawed sperm, a thermal resistance test (TRT) was executed. Samples preserved using Tris were in better condition compared with those preserved using ACP®, showing higher values for most assessments performed, including CASA and the TRT (P<0.05). After determining Tris to be the better of the two extenders, additional samples were thawed using different thawing rates (37°C/60 s, 55°C/7 s, 70°C/8 s). Sperm thawed at 37°C/60 s had the greatest preservation (P<0.05) of viability (54.1 ± 5.9%) and functional membrane integrity (43.2 ± 5.4%), and had higher values for various CASA parameters. In conclusion, we suggest the use of a Tris-based extender added to egg yolk and glycerol for the cryopreservation of epididymal sperm obtained from collared peccaries. In order to achieve better post-thawing sperm quality, we suggest that samples should be thawed at 37°C/60 s.

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Inclusion of Sericin in Tris Egg Yolk Extender Improves Bovine Sperm Quality during Cryopreservation (-196°C)

International Journal of Livestock Research ◽

10.5455/ijlr.20200424114516 ◽

2020 ◽

pp. 1

Author(s):

Tapasvi Patel ◽

Arjun Dhami ◽

Dinesh Chaudhari ◽

Kamlesh Hadiya

Keyword(s):

Sperm Quality ◽

Egg Yolk ◽

Bovine Sperm

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The use of butylated hydroxytoluene in cryopreservation of boar semen

Roczniki Naukowe Polskiego Towarzystwa Zootechnicznego ◽

10.5604/01.3001.0013.6966 ◽

2016 ◽

Vol 12 (2) ◽

pp. 21-28

Author(s):

Monika Trzcińska ◽

Magdalena Baryła

Keyword(s):

Dna Fragmentation ◽

Sperm Quality ◽

Annexin V ◽

Egg Yolk ◽

Butylated Hydroxytoluene ◽

Preimplantation Embryos ◽

Progressive Motility ◽

Boar Semen ◽

Boar Spermatozoa

The objective of the study was to determine the effect of butylated hydroxytoluene (BHT) on the quality and fertilizing capacity of frozen-thawed (FT) boar semen. Semen from five boars (36 ejaculates) was resuspended in lactose-egg yolk-glycerol extender supplemented with 0 (control), 1.0 (R1), 1.5 (R2) or 2.0 mM BHT (R3). Sperm quality was assessed based on motility (CASA; TM: total motility; PM: progressive motility), phosphatidylserine (PS) translocation across the plasma membrane (Annexin-V-FLuos Staining Kit) and DNA fragmentation (TUNEL Assay). The FT semen was also used for intrauterine artificial insemination (AI) of synchronized gilts. The fertilizing capacity of the FT semen was assessed on the basis of the gilt insemination rate and the number of morphologically normal embryos. The quality of the preimplantation embryos was determined by observing a TUNEL-positive reaction. The highest percentage of progressive motile and viable spermatozoa was noted in extender R3 (74.8 ±4.4% and 63.7 ±5.8%), as compared with the control (38.3 ±2.8% and 36.1 ±2.6%). The addition of BHT to the extender did not increase early apoptotic changes in the frozen-thawed spermatozoa with respect to the control. Irrespective of the variant of the extender, cryopreservation and thawing did not induce fragmentation in the boar spermatozoa. The highest number of morphologically normal embryos from inseminated gilts was observed in the case of semen cryopreserved in extender supplemented with 1.5 mM BHT. No significant differences were observed in DNA fragmentation in the expanded blastocysts from gilts inseminated with FT semen cryopreserved in the extenders analysed.

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Efficacy of egg yolk based and egg yolk free soya bean milk based extenders for cryopreservation of Zebu cattle and buffalo semen

Indian Journal of Animal Research ◽

10.18805/ijar.b-3453 ◽

2018 ◽

Author(s):

P. J. Chaudhary ◽

A. J. Dhami ◽

D. V. Chaudhari ◽

K. K. Hadiya ◽

J. A. Patel

Keyword(s):

Sperm Quality ◽

Membrane Integrity ◽

Egg Yolk ◽

Zebu Cattle ◽

Motile Sperm ◽

Comparative Efficacy ◽

Buffalo Semen ◽

Sample Technique ◽

Live Sperm ◽

Surti Buffalo

This study was undertaken on three mature bulls each of Gir cattle and Surti buffalo breeds to evaluate the comparative efficacy of egg yolk based standard TFYG (Tris-citrate-fructose-yolk-glycerol) extender and egg yolk free soybean based commercial extenders Optixcell® (IMV, France) and Andromed® (Minitube, Germany) under split-sample technique. The ejaculates (9/bull) were extended @ 100×106 sperm ml-1 with three extenders and frozen using biofreezer following 4 hr of equilibration. The pooled means of progressively motile sperm observed (irrespective of extenders) at initial, pre-freeze and post-thaw stage in Gir bulls semen were 76.53±0.53, 71.11±0.53 and 39.86±0.90% and in Surti buffalo 80.76±0.39, 74.65±0.45 and 40.35±1.07%, respectively. The corresponding values for live sperm were 75.64±0.76, 69.01±0.97 and 47.99±1.11 % for Gir and 80.90±0.45, 75.76±0.48 and 52.33±0.86 % for Surti buffalo; and those of intact acrosome 94.29±0.25, 90.29±0.27 and 79.29±0.33 % for Gir bulls, and 93.94±0.21, 89.94±0.23 and 78.95±0.26 % for Surti buffalo semen, respectively. The HOS reactive sperm at initial, pre-freeze and post-thaw stage were 76.18±0.74, 71.04±0.76 and 27.90±0.70 % for Gir, and 81.83±0.35, 76.47±0.39 and 27.83±0.68 % for Surti bulls, respectively. The overall mean post-thaw incubation (37°C) survival of spermatozoa observed at 60, 120 and 180 min were 28.40±0.91, 17.78±0.86 and 9.44±0.72% for Gir bulls semen, and 28.01±0.99, 18.40±1.01 and 10.51±0.93% for Surti buffalo semen, respectively. Optixcell was proved superior, and at par with TFYG, than the Andromed in maintaining greater motility, viability, morphology, acrosomal/plasma membrane integrity including post-thaw sperm longevity of cattle and buffalo spermatozoa with significant differences only in sperm motility and post-thaw longevity. The motile, live and HOST reactive sperm were significantly higher in buffalo semen than cattle at initial and pre-freeze stage, but not at post-thaw stage. The results showed that egg yolk free commercial Optixcell extender and egg yolk based TFYG extender were at par in terms of most of the sperm quality traits, hence any one of them can be preferred over Andromed for successful routine cryopreservation of cattle and buffalo semen.

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Comparative Quality Assessment of Buffalo (Bubalus bubalis) Semen Chilled (5°C) in Egg Yolk- and Soya Milk-Based Extenders

Reproduction in Domestic Animals ◽

10.1111/j.1439-0531.2011.01928.x ◽

2011 ◽

Vol 47 (4) ◽

pp. 596-600 ◽

Cited By ~ 11

Author(s):

AK Singh ◽

VK Singh ◽

BM Narwade ◽

TK Mohanty ◽

SK Atreja

Keyword(s):

Quality Assessment ◽

Egg Yolk ◽

Bubalus Bubalis ◽

Soya Milk

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Flow cytometric and microscopic evaluation of post-thawed ram semen cryopreserved in chemically defined home-made or commercial extenders

Animal Production Science ◽

10.1071/an13215 ◽

2015 ◽

Vol 55 (4) ◽

pp. 551 ◽

Cited By ~ 16

Author(s):

M. Emamverdi ◽

M. Zhandi ◽

A. Zare Shahneh ◽

M. Sharafi ◽

A. Akhlaghi ◽

...

Keyword(s):

Plasma Membrane ◽

Soybean Lecithin ◽

Sperm Quality ◽

Egg Yolk ◽

Mitochondrial Activity ◽

Flow Cytometric ◽

Microscopic Evaluation ◽

Open Question ◽

Ram Sperm

The present study was designed to determine the effect of three different extenders on ram sperm quality during a freeze–thawing procedure using flow cytometric and microscopic evaluations. Several in vitro qualitative analyses of post-thawed sperm parameters including motility and velocity parameters, plasma membrane functionality, total abnormality, capacitation status, acrosome integrity, mitochondrial activity and apoptosis features were considered. In the breeding season, seven ejaculates from each Zandi ram were collected routinely twice a week. Following semen collection, samples were pooled and equally divided into three aliquots. Each aliquot was diluted and frozen with one of the following extenders: (1) Tris-based extender containing 1.5% (w/v) soybean lecithin (TSL), as a chemically defined extender, (2) Bioxcell, a commercial soybean lecithin-based extender, and (3) Tris-based extender containing 20% (v/v) egg yolk (TEY). The results of the present study indicated no differences in total [TSL (55.8 ± 2.02%) vs TEY (50.2 ± 2.02%; P < 0.05)] and progressive motility of spermatozoa [TSL (26.2 ± 1.36%) vs Bioxcell (22.4 ± 1.36%; P < 0.05)]. Semen freezing by means of TSL resulted in a higher percentage of live spermatozoa (39.42 ± 1.81%) compared with TEY (29.17 ± 1.81%; P < 0.05), and a higher percentage of functional plasma membrane (50.8 ± 192%) compared with TEY (44 ± 1.92%) and Bioxcell (38.8 ± 1.92%; P < 0.05). The effect of extenders on sperm capacitation status showed that the percentage of post-thawed capacitated spermatozoa was higher in TEY (61.9 ± 1.48%) compared with that in TSL (56.6 ± 1.48%; P < 0.05). The evaluation of post-thawed spermatozoa indicated that the percentage of live spermatozoa with active mitochondria was higher in TSL (53.05 ± 2.31%) compared with Bioxcell (45.92 ± 2.31; P < 0.05) and the percentage of intact acrosome spermatozoa was higher in TSL (84.55 ± 2.51%) compared with TEY (74.91 ± 2.51%; P < 0.05). The use of TSL and Bioxcell extenders reduced the percentage of apoptotic spermatozoa (40.82 ± 2.07% and 42.22 ± 2.07%, respectively), compared with TEY (51.34 ± 2.07%; P < 0.05). Post-thawing dead spermatozoa were increased when semen was frozen by Bioxcell (25.69 ± 1.28%). The results of this study showed that TSL extender may provide stabile milieu and conditions for ram sperm cryopreservation compared with Bioxcell and TEY extenders. Whether TSL extender can improve the artificial insemination results remains, however, an open question.

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