Effect of sperm number and oxygenation state of the storage media on in vitro fertility of bovine sperm stored at ambient temperature

Krzyzosiak, Jacek; Molan, Peter C.; McGowan, Lindsay T.; Vishwanath, R.

doi:10.1016/s0093-691x(01)00490-3

Cited by 14 publications

(11 citation statements)

References 26 publications

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“…This difference of sperm motion could be possibly explained by differences in oxygen access for energetic metabolism. Although the respective effects of temperature and exposure to air could not be separated here, conservation of motility without air exposure is certainly dependent on glycolysis and possibly glycogen mobilization as suggested in other species [30][31][32] and in horses [3,33]. All these results suggest that equine sperm can use both aerobic and anaerobic pathways to survive, depending on the conditions of storage.…”

Section: Discussionmentioning

confidence: 77%

Temperatures from 4 to 15 °C are suitable for preserving the fertilizing capacity of stallion semen stored for 22 h or more in INRA96 extender

et al. 2012

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This study tested whether variable temperatures (from -0.5 to 15 °C) and air exposure could be used under laboratory and under field conditions to store stallion sperm diluted in extender INRA96 without loss of fertility. Experiment 1 (laboratory conditions) measured the effects of two 72 h storage conditions (5 °C with air vs. 15 °C without air). Experiment 2 (fixed field conditions) measured the effects of 22 h of storage without air in disposable containers maintained at four ambient temperatures (7 °C, 17 °C, 27 °C, 39 °C with semen at -0.5 °C to 3 °C, 4 °C to 7 °C, 8 °C to 10 °C, 12 °C to 15 °C, respectively). Per cycle pregnancy rate (PC) was measured after one artificial insemination (AI) in uterine body of 200×10(6) total spermatozoa, 7 h (Experiment 1) or 17 h (Experiment 2) before ovulation. In Experiment 1, PC was similar for both conditions (60% (n=40 cycles) vs. 63% (n=40), respectively, 5 stallions×8 cycles). Only velocity VCL and ALH were slightly higher at 15 °C. In Experiment 2, PC was reduced when ambient temperature was low (semen at -0.5 °C to 3 °C; PC=25%) rather than intermediate (semen at 4 °C to 7 °C; PC=53%) or high (semen at 8 °C to 10 °C; PC=50%) (4 stallions×8 cycles) (P=0.002). Sperm stored at -0.5 °C to 3 °C had lower acrosome integrity/responsiveness, similar membrane integrity (HOS test) and motilities, and higher VCL and ALH, than semen stored between 4 and 15 °C. These results demonstrate a wide tolerance of equine sperm to variable positive temperatures and air exposure for 22 h storage and more. However, temperatures close to 0 °C are detrimental for fertility.

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Section: Discussionmentioning

confidence: 77%

Temperatures from 4 to 15 °C are suitable for preserving the fertilizing capacity of stallion semen stored for 22 h or more in INRA96 extender

et al. 2012

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Section: Embryo Characteristics As Affected By Sperm Damagementioning

confidence: 99%

“…Thus, it is likely that the reduction in the proportion of blastocysts that were male caused by aging of sperm at 38.5 C reflects differential effects of aging on fertilizing ability of Y-bearing and X-bearing sperm. Energy store depletion, free radical damage, membrane changes or other aging-associated changes [44][45][46] could occur more rapidly for Y-bearing sperm, particularly if they are more active because of earlier capacitation.The reduction in male bias in sex ratio caused by aging at 38.5 C was not significant when sperm were aged at 40 C. Possibly, aging at 38.5 C affects Y-bearing sperm preferentially while aging at a higher temperature results in aging-associated changes in both Yand X-bearing sperm.The observation that aging of sperm can lead to changes in sex ratio of the resultant embryo points out the potential for changes in sperm function to effect the embryo formed by fertilization with that sperm. Nonetheless, despite nuclear and non-nuclear contributions of the sperm to the embryo [9], there was no evidence that the competence of the embryo to develop to the blastocyst stage was determined by aging at temperatures characteristic of normothermia or hyperthermia.…”

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confidence: 99%

Consequences for the Bovine Embryo of Being Derived from a Spermatozoon Subjected to Post-Ejaculatory Aging and Heat Shock: Development to the Blastocyst Stage and Sex Ratio

2009

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Abstract. The objective was to determine whether aging of sperm caused by incubation at normothermic (38.5 C) or heat shock (40 C) temperatures for 4 h prior to oocyte insemination affects sperm motility, fertilizing ability, competence of the resultant embryo to develop to the blastocyst stage and blastocyst sex ratio. In the first experiment, the percent of sperm that were motile was reduced by aging (P<0.001) and the reduction in motility was greater for sperm at 40 C compared to sperm at 38.5 C (P<0.01). In the second experiment, oocytes were inseminated with aged sperm. A smaller percent of oocytes fertilized with sperm aged at either temperature cleaved by Day 3 after insemination than oocytes fertilized with fresh sperm (P<0.05). There was no effect of sperm aging on the percent of oocytes or cleaved embryos that developed to the blastocyst stage. Aging of sperm before fertilization at 38.5 C reduced the percent of blastocysts that were male (P=0.08). In the third experiment, incubation of sperm at 38.5 C or 40 C for 4 h did not reduce fertilizing ability of sperm as determined by pronuclear formation at 18 h post insemination. In conclusion, aging of sperm reduced cleavage rate and the percent of blastocysts that were males but had no effect on the developmental capacity of the embryo. The effect of aging on cleavage rate may represent reduced motility and errors occurring after fertilization and pronuclear formation. Aging at a temperature characteristic of maternal hyperthermia had little additional effect except that polyspermy was reduced. Results indicate that embryo competence for development to the blastocyst stage is independent of sperm damage as a result of aging for 4 h at normothermic or hyperthermic temperatures. Key words: Aging, Bovine, Fertilization, Heat shock, Spermatozoa (J. Reprod. Dev. 55: [69][70][71][72][73][74] 2009) evelopmental competence of the mammalian embryo is dependent on genetic and non-genetic contributions from its parents [1][2][3][4][5][6]. Sperm could affect an embryo's competence for development if the timing of fertilization or early cleavage is delayed. For example, embryos produced by high-fertility bulls entered S-phase of the first cell cycle earlier and had a longer S-phase than those produced by low-fertility bulls [7]. In another study, spermatozoa from 50% of bulls identified as being of low fertility in artificial insemination studs experienced premature capacitation [8]. Damage to the macromolecular portions of the sperm that are incorporated by the embryo could also result in formation of embryos with reduced developmental competence. Among these sperm contributions are DNA, the centriole [9], and RNA [10]. Embryos fertilized with semen containing a high proportion of sperm with extensive DNA damage have reduced competence for development [11][12][13] but the importance of damage to the centriole or sperm RNA is not known.Damage to sperm can occur in the male reproductive tract or after deposition of sperm in the female. In bulls, for example, thermal...

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“…The main findings of this study were (i) storage of bull semen in INRA96 at o C is superior to semen stored at Although not assessed in the current study, plasma membrane integrity deteriorated quicker when stored in aerobic conditions compared to anaerobic or N gassed conditions [27], however, motility on Day 7 was significantly lower after storage under anaerobic conditions compared to aerobic or N 2 gassed storage conditions [27]. Therefore, it may be possible that diluents provides sufficient support for sperm motility over prolonged periods (up to 3 days of storage) regardless of the oxygenated state of the media and that the benefits of N 2 gassing are not observed by assessing motility alone.…”

Section: Discussionmentioning

confidence: 61%

Effect of storage temperature, nitrogen gassing and sperm concentration on the in vitro semen quality and in vivo fertility of liquid bull semen stored in INRA96

et al. 2018

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The aim of this study was to assess the effect of storage temperature, nitrogen (N) gassing and sperm concentration on in vitro characteristics and calving rate (CR) following artificial insemination (AI) of liquid bull semen stored in INRA96. In Experiment 1 the effect of liquid bull semen diluted in either N bubbled or non-bubbled INRA96 at a concentration of 5 × 10 sperm per 0.25 mL insemination dose and stored at 5 or 15 °C was assessed subjectively for total and progressive motility on Days 0, 1, 2, 3 and 4 post collection. In Experiment 2a, the effect of stored liquid semen at three sperm concentrations (3, 4 or 5 × 10 sperm per 0.25 mL insemination dose) on total and progressive motility was assessed subjectively on Days 0, 1 and 2 post collection. In Experiment 2b, the field fertility of liquid semen stored at ambient temperature at a concentration of 3, 4 or 5 × 10 sperm per 0.25 mL dose and inseminated on Days 1 or 2 post collection was assessed in comparison to frozen-thawed semen (total of n = 5742). In Experiment 1, total and progressive motility decreased with increased duration of storage (P < 0.01); however, there was no effect of N bubbling on motility on Days 0, 1, 2, 3 and 4 of storage. There was an effect of temperature on total and progressive motility, regardless of treatment, as semen stored at 15°C recorded higher motility values than semen stored at 5°C (P < 0.01). In Experiment 2a, there was no effect of sperm concentration on total or progressive motility on Days 0, 1 or 2 of storage. There was a linear decrease in motility with increased duration of storage (P < 0.01); however, there was no sperm concentration by day interaction. In Experiment 2b, there was an effect of sperm concentration on CR (P < 0.01); semen diluted to 3 and 4 × 10 sperm per dose resulted in a lower CR after 2 days of storage (41.1 and 44.7%, respectively) in comparison to frozen-thawed semen (55.2%) but did not differ to CR of semen diluted to 5 × 10 sperm per dose on Day 2 of storage. There was an effect of parity, fertility sub-index and days in milk (DIM) at AI on CR (P < 0.01). In conclusion, N bubbling and sperm concentration had no effect on in vitro sperm motility of liquid semen, but this study demonstrated a reduction in CR on Day 2 of storage at lower sperm concentrations in comparison to frozen-thawed semen.

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Effect of sperm number and oxygenation state of the storage media on in vitro fertility of bovine sperm stored at ambient temperature

Cited by 14 publications

References 26 publications

Temperatures from 4 to 15 °C are suitable for preserving the fertilizing capacity of stallion semen stored for 22 h or more in INRA96 extender

Temperatures from 4 to 15 °C are suitable for preserving the fertilizing capacity of stallion semen stored for 22 h or more in INRA96 extender

Consequences for the Bovine Embryo of Being Derived from a Spermatozoon Subjected to Post-Ejaculatory Aging and Heat Shock: Development to the Blastocyst Stage and Sex Ratio

Effect of storage temperature, nitrogen gassing and sperm concentration on the in vitro semen quality and in vivo fertility of liquid bull semen stored in INRA96

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