Artificial Insemination with Canine Semen Stored at a Low Temperature.

Tsutsui, Toshihiko; Tezuka, Tomoyuki; Mikasa, Yukiko; Sugisawa, Hirofumi; Kirihara, Nobuyuki; Hori, Tatsuya; Kawakami, Eiichi

doi:10.1292/jvms.65.307

Cited by 31 publications

(23 citation statements)

References 16 publications

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“…However, lower or higher concentration of sucrose did not significantly improve the progressive motility post-vitrification. Comparable results (60.7% of motility) was reported (Tsutsui et al, 2003) when the dog semen was chilled in egg yolk-**Tris at 4°C for over 4 days, but the spermatozoa lost their fertilizing capacity.…”

Section: Wwwintechopencomsupporting

confidence: 64%

Vitrification Technique - New Possibilities for Male Gamete Low-Temperature Storage

Isachenko¹,

Mallmann²,

Rahimi³

et al. 2012

Current Frontiers in Cryobiology

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Section: Wwwintechopencomsupporting

confidence: 64%

Vitrification Technique - New Possibilities for Male Gamete Low-Temperature Storage

Isachenko¹,

Mallmann²,

Rahimi³

et al. 2012

Current Frontiers in Cryobiology

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“…Sperm are very sensitive to temperature change, which easily affects sperm metabolism, motility and viability. High temperatures (around 38C) increase sperm metabolism and shorten survival, whereas, at 4C, sperm are able to remain viable for a long period of time by halting their movements [14]. The epididymides of animals are normally anaerobic, have low pH and do not contain carbohydrates as a source of energy; therefore, respiration and glycolysis are suppressed in sperm, and they remain viable for approximately one month [11].…”

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confidence: 99%

Influence of the Time between Removal and Cooling of the Canine Epididymis on Post-Thaw Caudal Epididymal Sperm Quality

et al. 2009

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ABSTRACT. The effects of the time period between canine epididymis removal and cooling on post-thaw caudal epididymal sperm quality were investigated. Sperm recovered from the epididymis stored for 6 hr at 4 or 20C exhibited similar motility. However, when the epididymis was stored for 12 hr or longer at 20C, sperm motility was significantly lower than that at 4C (p<0.01). The post-thawed qualities of sperm recovered from the caudal epididymides that had been stored at 20C for 0 or 6 hr and then at 4C for 24 hr after removal were not significantly different. Therefore, leaving the canine epididymis at 20C for up to 6 hr after its removal may have little effect on the post-thaw quality of recovered caudal epididymal sperm.KEY WORDS: canine, caudal epididymal sperm, frozen semen.J. Vet. Med. Sci. 71(6): 811-815, 2009 Many wild animals including canids are endangered. As artificial reproductive techniques to preserve these species, methods of gamete preservation have been investigated [1,5,7]. Gametes from male animals are generally collected by ejaculation. However, when animals die accidentally, sperm are recovered from the cauda epididymis, the sperm storage site [3,4,8].In a previous study [3], unilateral intrauterine insemination with sperm that was recovered from canine caudal epididymis in prostatic fluid (PF) and cryopreserved resulted in a high conception rate, 80.0%. We also showed that when caudal epididymal sperm were recovered after storage of the epididymis in sterile physiological saline at 4C for 48 hr, sperm motility was reduced, but intrauterine insemination with this post-thaw sperm again led to a high conception rate, 80.0% [4]. In these two experiments, the epididymis was kept at 4C immediately after removal, and the time period until the start of cooling after removal was not considered. However, when the bodies of male animals, such as wild animals, that have died accidentally are discovered, it is likely that several time have already passed under various temperature conditions, and so the epididymis can rarely be cooled immediately. This suggests that the time until cooling needs to be investigated.It has been reported that mouse caudal epididymal sperm shows a sperm motility of approximately 30% after 10 days of postmortem storage at 4C [6], but only shows a sperm motility of 10-15% after 24 hr of storage at room temperature [12]. Hishinuma et al. [2] reported that when the sika deer (Cervus nippon) epididymis is removed 4 hr postmortem and stored at 4C for 1-4 days, the caudal epididymal sperm motility is high, at approximately 50%, but, when removed 8-12 hr postmortem and stored at 4C for 1-4 days, the sperm motility is as low as 6.4%. These reports suggest that, in order to recover highly motile caudal epididymal sperm, it is important to store dead male animals immediately after death at 4C or to remove the epididymis as soon as possible and rapidly store it at 4C, thereby reducing the time of exposure to room temperature. However, in all the reported studies concerning...

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“…The semen was centrifuged (1,500 rpm, 5 min), and the seminal plasma was immediately removed. The precipitate was diluted with 20% egg yolk-tris/fructose-citrate extender (EYT-FC) [22] to adjust the sperm concentration to 200 × 10 6 /ml, followed by a 2nd dilution by dripping into it the same volume of EYT-FC containing 14% glycerol and 1.5% OEP for 10 min. The 1st and 2nd dilutions were performed at room temperature.…”

Section: Animalsmentioning

confidence: 99%

Effects of Liquid Nitrogen Vapor Sensitization Conditions on the Quality of Frozen-thawed Dog Spermatozoa

Hori

Odaka

Oba

et al. 2006

The Journal of Veterinary Medical Science

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ABSTRACT. The freezing conditions for preparation of frozen canine semen by the plunging method were investigated with regard to the period of sensitization in liquid nitrogen (LN 2 ) vapor and the height from LN 2 , and the semen qualities after thawing were compared with those of canine semen prepared by the simple freezer method previously reported by us. In the plunging method, 9 semen straws were prepared under the same conditions, horizontally kept at 5, 7, and 10 cm above the LN 2 surface in a styrene foam box for 5, 10, and 15 min, and then plunged into LN 2 . The semen qualities immediately after thawing were high in the 7 cm/10 min (cooling rate: -4 to -22°C/min) and 10 cm/15 min groups (cooling rate: -6 to -10°C/min). On comparison of frozen semen prepared by the plunging method (7 cm/10 min) with frozen semen prepared by the simple freezer method, sperm motility and viability were significantly higher for the frozen semen prepared by the plunging method. The cooling rate in freezing was higher for the simple freezer method (cooling rate: -6 to -50.9°C/min) than the plunging method. Based on these findings, horizontal placement of canine semen straws above LN 2 to reduce the temperature at a slow cooling rate of about -10°C/min, followed by plunging into LN 2 after sensitization for 10-15 min, provides good semen qualities after thawing. KEY WORDS: canine, cryopreservation, liquid nitrogen vapor, plunging method, simple freezer method.J. Vet. Med. Sci. 68(10): 1055-1061, 2006 Studies of frozen canine semen have not progressed because canine sperm is readily impaired by low-temperature sensitization compared to the sperm of other mammals [4]. It has recently been clarified that the addition of surfactants, such as Orvus ES paste (OEP: Nova Chemical Sales Inc., Scituate, MA, U.S.A.) [19][20][21] and Equex STM paste [12,14,16], to frozen semen extender containing egg yolk protects the acrosome cap after thawing, which prolongs the lifespan of sperm after thawing and increases the conception rate after artificial insemination. However, the preparation method of frozen canine semen varies among researchers, and factors that may affect the qualities of frozen semen after thawing, such as the diluent composition [2,6,13,16,17], dilution rate [9], cryoprotectant concentration [2], packaging (straw thickness) [8], freezing method (cooling rate) [1,8,10,11,15,16,18,23], and thawing method (warming rate) [8,10,23], have been investigated.For freezing, we previously used a conventional simpletype quick LNG freezer (FA-1652, Fujihira Industry Co., Ltd., Tokyo, Japan) and obtained a high conception rate (90%) by intrauterine insemination using a thawed semen preparation [19][20][21]. This instrument, developed for the preparation of frozen bovine semen, is capable of preparing a large amount of frozen semen in a single operation. However, the opportunity to prepare such large amounts of frozen canine semen is rare. In another simple method, called the plunging method, semen is exposed to liquid nitrogen (LN 2 ) vap...

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Artificial Insemination with Canine Semen Stored at a Low Temperature.

Cited by 31 publications

References 16 publications

Vitrification Technique - New Possibilities for Male Gamete Low-Temperature Storage

Vitrification Technique - New Possibilities for Male Gamete Low-Temperature Storage

Influence of the Time between Removal and Cooling of the Canine Epididymis on Post-Thaw Caudal Epididymal Sperm Quality

Effects of Liquid Nitrogen Vapor Sensitization Conditions on the Quality of Frozen-thawed Dog Spermatozoa

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