Sperm fertility and viability following 48h of refrigeration: Evaluation of different extenders for the preservation of bull semen in liquid state

Crespilho, André Maciel; Nichi, Marcílio; Guasti, Priscilla Nascimento; Freitas-Dell’Aqua, C.P.; Filho, M. F. Sá; Maziero, R. R. D.; Dell’Aqua, J.A.; Papa, Frederico Ozanam

doi:10.1016/j.anireprosci.2014.02.020

Cited by 33 publications

(25 citation statements)

References 43 publications

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“…In consideration of differences in FTAI synchronization protocols and female reproductive status, the egg yolk‐based extenders have so far proven superior in achieving high pregnancy rates (Crespilho et al., ) in lieu of low‐density lipoprotein support of the sperm membrane in addition to glycerol (Vera‐Munoz et al., ) for electroejaculated B. indicus beef cattle. Our mean pregnancy rates in both treatment groups were comparable to earlier studies using FTAI in Nellore cattle with semen collected using electroejaculation and extended in a modified Tris‐egg yolk or commercial extender (Borges‐Silva et al., ; Crespilho et al., ; Papa et al., ). There was a significant reduction in pregnancy rate to FTAI using chilled‐stored semen compared to conventional cryopreserved semen, which therefore establishes probable extrinsic factors that adversely affect spermatozoa in liquid storage.…”

Section: Discussionsupporting

confidence: 84%

“…Chilled storage has also been shown to decrease in motility, plasma membrane integrity and acrosome integrity decreases with duration of chilled storage of bull semen (Nair, Brar, Ahuja, Sangha, & Chaudhary, ). One recent study reported a decline in both in vitro sperm function and pregnancy rate with 48‐h chilled‐stored semen compared to cryopreserved semen (Crespilho et al., ). Conversely, these measures are also reported to be stable for the first 48 hr in some extenders, with the exception of acrosome integrity (Vera‐Munoz et al., ).…”

Section: Discussionmentioning

confidence: 99%

“…Ascertaining sperm longevity must be taken into account for FTAI, as insemination is performed at a fixed‐time point, based on average ovulation time, rather than in response to individual cyclic signs. In order to increase sperm longevity, a number of recent studies have started to incorporate chilled semen storage of semen in an effort to further improve pregnancy outcome of FTAI in B. indicus breeds (Borges‐Silva et al., ; Crespilho, Papa, de Paula Santos, & de Sa Filho, ; Crespilho et al., ; Papa et al., ). The advantages include a presumptive higher viability and longevity than cryopreserved semen, the opportunity to reduce the semen dose (low dose insemination and producing more doses per collected ejaculate), the availability of on‐farm bulls and decrease cost associated with acquisition, handling and storage of semen; namely attributed to the removal of the need to access liquid nitrogen, electricity and other specialized technical equipment (Schuh, ).…”

Section: Introductionmentioning

confidence: 99%

“…SATAKE ET Al. improve pregnancy outcome of FTAI in B. indicus breeds (Borges-Silva et al, 2015;Crespilho, Papa, de Paula Santos, & de Sa Filho, 2012a;Crespilho et al, 2014;Papa et al, 2015). The advantages include a presumptive higher viability and longevity than cryopreserved semen, the opportunity to reduce the semen dose (low dose insemination and producing more doses per collected ejaculate), the availability of on-farm bulls and decrease cost associated with acquisition, handling and storage of semen; namely attributed to the removal of the need to access liquid nitrogen, electricity and other specialized technical equipment (Schuh, 1992).…”

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confidence: 99%

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Investigation of in vitro measurable sperm attributes and their influence on electroejaculated bull semen with a fixed‐time artificial insemination protocol in AustralianBos indicuscattle

Satake

Edwards

Tutt

et al. 2017

Reprod Domestic Animals

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Increasing use of fixed-time artificial insemination (FTAI) in beef cattle production has presented an opportunity for the use of fresh or chilled semen as an alternative to standard cryopreserved semen. The objective of this study was to examine in vitro sperm function and pregnancy rate of electroejaculated semen, chilled and stored for 48 hr, compared to conventionally cryopreserved semen with an optimized FTAI protocol in Brahman cattle. Semen from three Brahman bulls was collected, and aliquots were extended in either chilled (at 5°C) or frozen (LN ) in a Tris-egg yolk extender base with 2.4% or 7.0% glycerol, respectively. Semen samples were assessed 48 hr after collection or post-thaw and warming, for sperm motility, in vitro sperm function and fertilizing ability, and used in a FTAI programme. The overall pregnancy rates was significantly different (p < .01) after FTAI with frozen (n = 173; 53.2%) and chilled semen (n = 174; 31.6%). In contrast, the in vitro sperm assessment showed that the chilled semen had significantly faster motility (p < .05), a higher proportion of progressively motile spermatozoa (p < .05), with significantly higher proportions of acrosome intact, viable spermatozoa (p < .01). This study showed that reasonable pregnancy rates in Brahman cattle can be achieved using FTAI with chilled semen collected using electroejaculation and stored for up to 48 hr. However, improvements in semen extenders are required in consideration of semen collection method to improve the longevity of sperm fertilizing ability to significantly increase FTAI output using chilled storage of bull semen.

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Section: Discussionsupporting

confidence: 84%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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Investigation of in vitro measurable sperm attributes and their influence on electroejaculated bull semen with a fixed‐time artificial insemination protocol in AustralianBos indicuscattle

Satake

Edwards

Tutt

et al. 2017

Reprod Domestic Animals

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“…Cooled bull semen is used mainly in fixed-time artificial insemination programmes (Crespilho et al, 2014), because of its limited shelf life (Vishwanath & Shannon, 2000;Verberckmoes et al, 2004). However, the advantage of using fresh semen includes the ability to inseminate a large number of animals in a short time (Bucher et al, 2009) with relatively low sperm numbers per insemination, high sire utilization, inexpensive storage, and ease of use in the field (Vishwanath & Shannon, 2000;Verberckmoes et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Viability of bull semen extended with commercial semen extender and two culture media stored at 24 °C

Raseona¹,

Ajao²,

Nethengwe³

et al. 2017

SA J. An. Sci.

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The aim of this study was to evaluate the viability of bull spermatozoa diluted with commercial semen extender and two culture media stored at controlled room temperature (24 °C) for 72 hours. Two Nguni bulls were used for semen collection with the aid of an electro-ejaculator. After macroscopic evaluation, semen was pooled and aliquoted randomly into Triladyl, modified Ham's F10, and TCM-199 culture media, and then stored at 24 °C. Sperm motility parameters, morphology, and viability were analysed with computer aided sperm analysis (CASA) after 0, 24, 48 and 72 hours. The study was replicated four times, and data were analysed using analysis of variance (ANOVA). Triladyl had significantly higher sperm viability rate (41.3%) and total motility rate (96.3%) for 72 hours than modified Ham's F10 (86.8%; 26.5%) and TCM-199 (76.7%; 25.0%) culture media. Ham's F10 had higher progressive motility rate (37.8%) than the other extenders TCM-199 (31.7%) and Triladyl (23.4%). There was no significant difference in viability rate between Ham's F10 (26.5%) and TCM-199 (25.0 %) after 72 hours' storage at 24 °C. Furthermore, no significant difference was observed in total sperm abnormalities, except for reacted acrosomes and absent tails, between the two Nguni bulls. Nguni semen can be preserved in Triladyl or modified Ham's F10 and TCM-199 culture media, stored at 24 °C, and stay viable for 72 hours.

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Evaluation of cooling and freezing systems of bovine semen

Dias

Campanholi

Rossi

et al. 2018

Animal Reproduction Science

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Semen cryopreservation comprises different steps, among them are the cooling and freezing rates which significantly influence the quality of thawed sperm. Different systems with variable freezing rates are used for freezing bull semen in the field, with a consequence of variable success rates. The objective of this study was to compare different systems for freezing bull semen in the field. Five cooling methods of semen and two methods for the subsequent freezing phase (5 × 2 factorial scheme) were used. Two to four ejaculates were collected from 12 bulls with an electroejaculator. The ejaculates were diluted in BotuBov® to a concentration of 50 × 10 6 spermatozoa/mL in 0.5-mL straws. After dilution, the straws were cooled to 5°C in five cooling systems: TK 4000® at a cooling rate of −0.25°C/min (R1); TK 4000® at a rate of −0.5°C/min (R2); Minitube® refrigerator at a rate of −2.8°C/min (R3); Botutainer® at a rate of −0.65°C (R4), and domestic refrigerator at a rate of −2.0°C/min (R5). After stabilization at 5°C for 4 h, these straws were then submitted to two freezing systems: TK 4000® at a freezing rate of −15°C/min (C1) and Styrofoam box with liquid nitrogen at a rate of −19°C/min (C2). Sperm kinetics were evaluated by computer-assisted sperm analysis at four time points: in fresh semen, after cooling, post-thawing, and after the rapid thermal resistance test (TRT). In addition, plasma and acrosomal membrane integrity, mitochondrial potential and intracellular H 2 O 2 were analyzed after thawing by flow cytometry. The R1, R2 and R4 cooling systems were the most efficient in preserving sperm viability, membrane integrity and intracellular H 2 O 2. Samples frozen in the C1 system exhibited better post-thaw and post-TRT kinetics than C2 samples. In conclusion, slower cooling curves in conjunction with a constant freezing rate obtained with the programmable unit were more efficient for freezing bull semen in the field.

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Sperm fertility and viability following 48h of refrigeration: Evaluation of different extenders for the preservation of bull semen in liquid state

Cited by 33 publications

References 43 publications

Investigation of in vitro measurable sperm attributes and their influence on electroejaculated bull semen with a fixed‐time artificial insemination protocol in AustralianBos indicuscattle

Investigation of in vitro measurable sperm attributes and their influence on electroejaculated bull semen with a fixed‐time artificial insemination protocol in AustralianBos indicuscattle

Viability of bull semen extended with commercial semen extender and two culture media stored at 24 °C

Evaluation of cooling and freezing systems of bovine semen

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