Effects of extender and equilibration time on post-thaw motility and membrane integrity of cryopreserved Gyr bull semen evaluated by CASA and flow cytometry

Leite, Ticiano Guimarães; Filho, Vicente Ribeiro do Vale; Arruda, Rubens Paes de; Andrade, André Furugen Cesar de; Emerick, Lucas Luz; Zaffalon, Fabiane Gilli; Martins, Jorge André Matias; Andrade, Venício José de

doi:10.1016/j.anireprosci.2010.04.005

Cited by 97 publications

(89 citation statements)

References 38 publications

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“…Our findings are in agreement with Shahverdi et al (2014) Leite et al (2010) determined 240 min as the most appropriate length of equilibration period in comparison to non-equilibration and 120 min equilibration, in accord with our study. The non-equilibration gave the lowest values of total and progressive sperm motility, and higher percentage proportion of sperm with intact plasma and acrosomal membranes, with no significant differences between different extenders used.…”

Section: Discussionsupporting

confidence: 94%

“…After filling, straws were divided into 3 groups (n = 20 insemination doses per sire at least), immediately placed into a cooling box, cooled at an average speed of 0.2°C per min to 4-5°C, and equilibrated (Camara et al 2011). According to pre-defined methodologies, equilibration lengths of 30 min (Leite et al 2010), 120 min (Shahverdi et al 2014), and 240 min (Januskauskas et al 1999) were applied and tested. Subsequently, equilibrated straws were divided into 2 groups (n = 10 insemination doses per sire per equilibration length at least) and freezed using 2 freezing curves providing the highest spermatozoa motility after thawing .…”

Section: Methodsmentioning

confidence: 99%

“…Equilibration as the total period of spermatozoa contact with a cryoprotectant prior to freezing helps keep spermatozoa membrane integrity as well doi: 10.17221/23/2016-CJAS as their survival (Leite et al 2010). Extenders used serve as protection during cooling and freezing (Linhartova et al 2014) and preserve spermatozoa motility after subsequent thawing (Muchlisin et al 2015).…”

Section: Introductionmentioning

confidence: 99%

“…Nevertheless, the most suitable interactions of extender vs equilibration length still are not determined sufficiently (Shahverdi et al 2014). Leite et al (2010) recorded only limited interactions between the extender used and the equilibration length. Dhami et al (1992) recommended 30 min to 24 h equilibration prior to freezing, while Muino et al (2007) determined optimal length of equilibration from 4 to 18 h. Selected studies indicate that 18 h or overnight equilibrations prior to freezing are the most appropriate equilibration lengths in order to increase quality of the insemination doses produced (Shahverdi et al 2014).…”

Section: Introductionmentioning

confidence: 99%

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Equilibration and freezing interactions affecting bull sperm characteristics after thawing

Doležalová¹,

Stádník²,

Biniová³

et al. 2016

CZECH J ANIM SCI

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ABSTRACT:The objective was to determine effects of equilibration length and freezing curve type as well as their interactions on motility and live spermatozoa proportion in bull sperm after thawing. The ejaculates of 6 sires were repeatedly collected. Fresh semen was diluted with one extender and divided into 3 groups equilibrated for 30, 120, and 240 min. Subsequently, half straws of each group were frozen using standard 3-phase or 2-phase freezing curve differing in the rate of temperature decrease. Th e spermatozoa motility (M) was evaluated immediately after thawing and at 30, 60, 90, and 120 min of thermodynamic test (TDT). Live spermatozoa proportion was evaluated after thawing and at the end of TDT. Average of spermatozoa motility (AM), decrease of spermatozoa motility (MD), average proportion of live spermatozoa (ALS), and decrease of live spermatozoa proportion (DLS) through the TDT were calculated. Significant inter-sire differences in AM (0.45-17.0%; P < 0.05-0.01), MD (0.76-12.57%; P < 0.05-0.01), and ALS (0.99-23.8%; P < 0.01) were detected. The longest equilibration ensured the highest M during TDT and AM (+2.72 and +4.58%; P < 0.05-0.01), however higher MD (+4.06%; P < 0.01) compared to standard length as well. Straws freezed using 2-phase curve achieved higher M through TDT, AM (+7.3%; P < 0.01) as well as ALS (+11.77%; P < 0.01). The 2-phase curve presented higher M compared to the 3-phase freezing curve within all equilibration lengths. Significant differences in AM, MD,; P < 0.05-0.01) between equilibration length vs freezing curve interactions were determined. Results document the importance of equilibration length, freezing curve, and their interaction effect on live spermatozoa proportion and sperm motility after thawing as well as necessity of individual conditions for bulls semen processing and insemination doses production.

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Section: Discussionsupporting

confidence: 94%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Equilibration and freezing interactions affecting bull sperm characteristics after thawing

Doležalová¹,

Stádník²,

Biniová³

et al. 2016

CZECH J ANIM SCI

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“…In our study, E2 compared to other equilibration times has resulted in significantly higher post-thaw sperm PM, RV, VAP, VSL, VCL, STR, LIN, SV-PMI, MMP, and V-IACR. Similar to our results, some studies have reported improved post-thaw sperm STR and LIN with 4 h equilibration time in buffalo (Kumar et al, 2015) and cattle (Celeghini et al, 2008;Leite et al, 2010). Tuli et al (1981) have also reported higher sperm motility in buffalo with 4 h equilibration compared to 2 and 6 h equilibration times.…”

Section: Discussionsupporting

confidence: 91%

Effect of equilibration times, freezing, and thawing rates on post-thaw quality of buffalo (Bubalus bubalis) bull spermatozoa

Shah

Andrabi²,

Qureshi

2016

Andrology

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SUMMARYThe effects of equilibration times (E1, 2 h; E2, 4 h; E3, 6 h), freezing rates (FR1, manual, 5 cm above liquid nitrogen (LN 2 ) for 10 min, plunging in LN 2 ; FR2, programmable ultra-fast, holding at +4°C for 2 min, from 4 to À10°C at À10°C/min, from À10 to À20°C at À15°C/min, from À20 to À120°C at À60°C/min, holding at À120°C for 30 sec, plunging in LN 2 ), and thawing rates (T1, 37°C for 30 sec; T2, 50°C for 15 sec; T3, 70°C for 7 sec) were evaluated on quality of buffalo bull spermatozoa. Progressive motility (%), rapid velocity (%), average path velocity (VAP, lm/s), straight line velocity (VSL, lm/s), and mitochondrial transmembrane potential (%) were higher (p < 0.05) with E2, FR2, and T3 compared to other groups. Sperm curved line velocity (VCL, lm/s) was higher (p < 0.05) with E2 and FR2 compared to other groups. Sperm straightness (%) and linearity (LIN, %) were higher (p < 0.05) with E2 compared to other groups. Sperm LIN was affected (p < 0.05) with T3 compared to other groups. Supravital-plasma membrane integrity (%), viability and acrosome integrity (%) of spermatozoa were higher (p < 0.05) with E2 and FR2 compared to other groups. Sperm DNA integrity (%) was higher (p < 0.05) with FR2 and T1 compared to other groups. We concluded that inclusion of 4 h-equilibration time, programmable ultra-fast freezing rate, and rapid thawing at 70°C for 7 sec in cryopreservation protocol improves the post-thaw quality of buffalo bull spermatozoa.

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Effect of Cryopreservation and Packaging System on Sperm Motility and Fertility of Striped Catfish

et al. 2021

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The Orinoco Striped Catfish Pseudoplatystoma orinocoense is a highly valued commercial‐capture species in Colombia. Although this species was considered endangered and is now in a recovery situation, there are still no cryopreservation studies for its milt. Therefore, the objective of this study was to evaluate the effects of different cryopreservation media and two packaging systems on the sperm motility and fertilization capacity of Striped Catfish. A first experiment was conducted to test the effects of two permeating cryoprotectants (10% dimethyl sulfoxide and 12% methanol) combined with glucose at two concentrations (5.5% and 10%) and with two membrane stabilizers (12% egg yolk and 5% whole milk powder) on the post‐thaw motility of milt samples that were packed into 0.5‐mL straws. The cryopreservation media with the best results in the first experiment were selected for a second experiment in which we evaluated 5‐mL macrotubes as a packaging system. In this case, the response variables were sperm motility and fertility rate. In both experiments, milt with sperm motility that exceeded 90% was diluted at a 1:6 (volume basis) ratio in each of the two cryopreservation media. The diluted samples were packed, equilibrated (for 10 min), then frozen in a nitrogen‐vapor dry shipper for 30 min, and finally stored in liquid nitrogen until they were evaluated. Each factor and its interactions showed a significant effect (P < 0.001) on post‐thaw sperm motility in experiment 1. The best protection during cryopreservation was obtained when 12% methanol supplemented with 5.5% glucose was used as a permeating cryoprotectant, yielding an average motility of 33.3 ± 5.1% (mean ± SD) and 64 ± 5.4% in the 0.5‐ and 5‐mL packaging systems, respectively. Similarly, the highest fertility rate (85%) was recorded under this treatment, with no significant difference (P > 0.05) from the fresh milt. Therefore, Striped Catfish milt can be cryopreserved in 5‐mL macrotubes by using 12% methanol and 5.5% glucose, without the addition of membrane stabilizers.

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Effects of extender and equilibration time on post-thaw motility and membrane integrity of cryopreserved Gyr bull semen evaluated by CASA and flow cytometry

Cited by 97 publications

References 38 publications

Equilibration and freezing interactions affecting bull sperm characteristics after thawing

Equilibration and freezing interactions affecting bull sperm characteristics after thawing

Effect of equilibration times, freezing, and thawing rates on post-thaw quality of buffalo (Bubalus bubalis) bull spermatozoa

Effect of Cryopreservation and Packaging System on Sperm Motility and Fertility of Striped Catfish

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