Effect of cooling rates on post-thaw sperm motility, membrane integrity, capacitation status and fertility of dairy bull semen used for artificial insemination in sweden

Januškauskas, Aloyzas; Gil, J.; Söderquist, L.; Hrd, M.G.M.; Hrd, M.Ch.; Johannisson, Anders; Rodriguez‐Martinez, Heriberto

doi:10.1016/s0093-691x(99)00159-4

Cited by 87 publications

(55 citation statements)

References 40 publications

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“…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%

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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: Methodsmentioning

confidence: 99%

“…Freezing was performed using the controlled freezing methodology Direct Freezing in a freezer box Digitcool ® (IMV Cryo Bio System, L'Aigle, France). Muino et al 2007) and the 2-phase freezing curve described by Januskauskas et al (1999) and Gil et al (2000) were applied.…”

Section: Methodsmentioning

confidence: 99%

Equilibration and freezing interactions affecting bull sperm characteristics after thawing

Doležalová¹,

Stádník²,

Biniová³

et al. 2016

CZECH J ANIM SCI

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“…As membrane fluidity increases, more M540 is able to intercalate into the membrane, thereby acting as a useful marker for membrane destabilization [5,11,[35][36][37][38][39]. Moreover, increased M540 staining occurs within a few minutes after exposure of boar sperm to bicarbonate, and much earlier than TP staining is able to detect capacitation in stallion sperm [5,35].…”

Section: Introductionmentioning

confidence: 99%

“…Moreover, increased M540 staining occurs within a few minutes after exposure of boar sperm to bicarbonate, and much earlier than TP staining is able to detect capacitation in stallion sperm [5,35]. M540 staining indicates an early stage of sperm capacitation in the boar [5,11], bull [38,39], and stallion [35] but has yet to be validated for the dog. Dog prostatic fluid (PF) contains components that mask the progesterone receptors on the sperm plasma membrane, postponing initiation of the capacitation process until removal [10,40,41].…”

Section: Introductionmentioning

confidence: 99%

Validation of merocyanine 540 staining as a technique for assessing capacitation-related membrane destabilization of fresh dog sperm

Steckler¹,

Stout

Durandt

et al. 2015

Theriogenology

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The aim of this study was to determine whether flow cytometric evaluation of combined merocyanine 540 and Yo-Pro 1 staining (M540-YP) would identify viable dog sperm that had undergone membrane stabilization known to be associated with capacitation in other species, and whether such destabilization is detected earlier than when using the tyrosine phosphorylation and ethidium homodimer stain combination (TP-EH) with epifluorescence microscopy. Semen from nine dogs was collected and incubated in parallel in bicarbonate-free modified Tyrode"s medium (-BIC), in medium containing 15 mM bicarbonate (+BIC), in dog prostatic fluid (PF), and in phosphate buffered saline (PBS). Aliquots for staining were removed at various time points during incubation of up to 6 hours. Staining with M540-YP allowed the classification of dog sperm as viable without destabilized membranes, viable with 2 destabilized membranes, non-viable without destabilized membranes or non-viable with destabilized membranes. The percentage of viable sperm detected using EH (83.5 ± 1.37%; mean ± SEM) was higher than when using YP (66.7 ± 1.37%: P < 0.05; n=54 semen samples). On the other hand, M540-YP identified a higher percentage of viable sperm with destabilized membranes than TP-EH (75 ± 1.76% vs. capacitation was recorded during incubation in PBS. We conclude that YP identifies sperm committed to cell death earlier than EH, and that the M540-YP stain combination identifies membrane destabilization known to be associated with capacitation in other species earlier than the TP-EH stain combination.

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“…light or fluorescent microscopy combined with vital stains (Brito et al, 2003), and flow cytometry (Januskauskas et al, 1999;Hallap et al, 2004). One of the simplest methods to evaluate the plasmalemma of sperm cells is the hypo-osmotic swelling (HOS) test.…”

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confidence: 99%

Relationships between the results of hypo-osmotic swelling tests, sperm motility, and fertility in Estonian Holstein dairy bulls

Padrik¹,

Hallap²,

Kaart³

et al. 2012

CZECH J ANIM SCI

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ABSTRACT:As an attempt to find an inexpensive and simple laboratory method for artificial insemination (AI) bull semen quality assessment, the osmotic resistance of spermatozoa was measured using the hypoosmotic swelling (HOS) test, developed by Jeyendran et al. (1984) (labelled HOS-1), and its modifications (HOS-2, HOS-3), with decreased osmotic pressure aimed at challenging sperm survival ability. The test results were benchmarked against sperm viability measurements performed using the Computerized Motility Analyzer (CMA), and field fertility was calculated as non-return rate (NRR). Two age groups of Estonian Holstein bull sires were included in this study to test possible age effect on semen quality parameters. The HOS-1 test in fresh bull semen correlated well with sperm general motility (GMot) (r = 0.63, P < 0.001 at batch level and r = 0.77, P < 0.001 at bull level) as well as with progressive motility (PMot) in frozen-thawed (FT) semen (r = 66, P < 0.001 at batch level and r = 0.81, P < 0.001 at bull level), which makes the test suitable for the prediction of post-thaw semen quality. However, the HOS-2 and HOS-3 values in FT semen had high correlations with NRR (r = 0.65, r = 0.66, P < 0.001 at batch level and r = 0.63, r = 0.71, P < 0.01 at bull level), which was comparable to those between GMot and NRR or PMot and NRR. A combination of motility parameters and the results of the HOS-1 and HOS-3 tests provided a good model for predicting the potential fertility of bull semen. Values of sperm membrane post-thaw intactness, assessed using HOS-2, as well as of sperm motility measurements were higher in mature bulls compared to those in young bulls. Short conclusion: different modifications of the hypo-osmotic swelling test are useful for routine bovine semen quality assessment at AI stations.Keywords: bull fertility; semen quality; sperm membrane intactness; bull age Laboratory analyses have been used over many decades to evaluate semen quality. Despite the fact that none of the single assays developed provide results that consistently and highly correlate with fertility, the laboratory evaluation of semen samples remains an important procedure for the artificial insemination (AI) industry to eliminate low fertility bulls or semen from being used in artificial insemination programmes (Graham and Mocé, 2005).Post-thaw viability and fertility of cryopreserved sperm are often reduced due to accumulated cellular damage during the cryopreservation phases (Muiño et al., 2008). Decrease in temperature, cold shock, and intracellular ice formation can affect sperm plasma membrane, acrosomal and mitochondrial membrane integrity (Thomas et al., 1998;Defoin et al., 2008), and cause the loss of intracellular components (Graham and Mocé, 2005), which can initiate cell death.Sperm membrane integrity can be evaluated by several methods, e.g. light or fluorescent microscopy combined with vital stains (Brito et al., 2003), and flow cytometry (Januskauskas et al.,

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Effect of cooling rates on post-thaw sperm motility, membrane integrity, capacitation status and fertility of dairy bull semen used for artificial insemination in sweden

Cited by 87 publications

References 40 publications

Equilibration and freezing interactions affecting bull sperm characteristics after thawing

Equilibration and freezing interactions affecting bull sperm characteristics after thawing

Validation of merocyanine 540 staining as a technique for assessing capacitation-related membrane destabilization of fresh dog sperm

Relationships between the results of hypo-osmotic swelling tests, sperm motility, and fertility in Estonian Holstein dairy bulls

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