Electroejaculation and semen analysis and freezing in the giant panda (Ailuropoda melanoleuca)

Platz, C. C.; Wildt, David E.; Howard, Jo Gayle; Bush, M.

doi:10.1530/jrf.0.0670009

Cited by 44 publications

(40 citation statements)

References 6 publications

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“…Seminal volume (2.8^0.4 ml) and sperm concentration (1.1^0.3 £ 10 9 sperm/ml), initial motility (76.1^4.4%), forward progression rating (3.4^0.2) and acrosomal integrity (93.6^1.7% intact) for giant panda ejaculates collected here were consistent with previous reports (Platz et al 1983, Chen et al 1994. Most spermatozoa were morphologically normal (84.2^3.0%), although many (74.9^8.2%) displayed the interesting abaxial attachment trait for the head/midpiece region as noted previously .…”

Section: Resultssupporting

confidence: 91%

“…Deep anaesthesia was maintained with 0 -5% isofluorane gas, as needed. The electroejaculation technique has been described (Platz et al 1983, Howard 1993, 1999) and briefly relied on using a 2.6 cm diameter rectal probe with three longitudinal, stainless-steel electrodes and a 60 Hz sine wave stimulator (P.T. Electronics, Boring, OR, USA).…”

Section: Semen Collection/analysismentioning

confidence: 99%

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Acrosomal integrity and capacitation are not influenced by sperm cryopreservation in the giant panda

Spindler¹,

Huang²,

Howard³

et al. 2004

Reproduction

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Sperm cryopreservation and artificial insemination are important management tools for giant panda breeding and the preservation of extant genetic diversity. This study examined the influence of freeze-thawing on sperm function, specifically capacitation. Sperm from nine giant pandas were assessed before and after rapid ( 2 40 and 2 100 8C/min) cryopreservation by incubation in HEPES-buffered Ham's F10 medium with and without the capacitation accelerators, 3-isobutyl-1-methylxanthine (IBMX) and dibutyryl cyclic AMP (dbcAMP). At 0, 3 and 6 h of exposure, aliquots were assessed for sperm motility traits and capacitation, defined as the proportion of sperm with intact acrosomes following exposure to solubilised zonae pellucidae (ursid or felid) or calcium ionophore subtracted from the proportion of sperm with intact acrosomes before exposure. Although mean6 S.E.M. sperm motility post-thaw (56.1 6 3.9% at 0 h) was less (P < 0.05) than pre-freeze (71.7 6 6.0%), there was no difference (P > 0.05) in the proportion of acrosome-intact sperm (fresh, 93.0 6 1.7% versus cryopreservedthawed, 81.7 6 4.7% at 0 h). Incidence of capacitation was greater (P < 0.05) in fresh sperm incubated with capacitation accelerators IBMX and dbcAMP (9 h: 50.9 6 1.1) compared with fresh sperm incubated without accelerators (9 h: 41.2 6 1.1%). Frozen-thawed sperm preincubated without accelerators underwent capacitation (49.6 6 1.1%) to a greater extent (P < 0.05) compared with these fresh counterparts. Thawed samples with (9 h: 45.9 6 1.4%) and without accelerators (9 h: 41.2 6 1.1%) did not differ (P > 0.05) during the 9-h incubation. We conclude that giant panda spermatozoa (1) undergo capacitation in vitro with or without chemical accelerators and (2) withstand a rapid cryopreservation protocol, including retaining normal acrosomal integrity and functional capacitation ability.

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Section: Resultssupporting

confidence: 91%

Section: Semen Collection/analysismentioning

confidence: 99%

Acrosomal integrity and capacitation are not influenced by sperm cryopreservation in the giant panda

Spindler¹,

Huang²,

Howard³

et al. 2004

Reproduction

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“…There was no significant correlation between both motility and viability of frozen-thawed spermatozoa and any characteristics of fresh semen (Tables 2 and 3). The mean percentage of motile frozen-thawed spermatozoa in the present study (43 ± 5%) was comparable to that collected by electroejaculation in giant panda (< 55%) [14], red-wolf (< 40%) [5] and Tibetan black bear (0-60%) [9], that collected by epididymal mincing in Cantablic brown bear (25%) [2], and that collected by artificial vagina in dog (< 50%) [8,12].…”

supporting

confidence: 76%

Cryopreservation of the Semen Collected by Electroejaculation from the Hokkaido Brown Bear(Ursus arctos yesoensis).

Ishikawa

Matsui

Sakamoto³

et al. 2002

The Journal of Veterinary Medical Science

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ABSTRACT. Four adult Hokkaido brown bears were used as semen donors, and semen characteristics were examined before freezing and after thawing. A total of 10 electroejaculates were diluted with Tris-egg yolk extender and cooled to 4°C over 90 min. Spermatozoa were equilibrated with 4.7% glycerol for 80 min. Semen packed in 0.25 ml plastic straws were frozen with liquid nitrogen vapor. Percentages (mean ± SD) of motile and live sperm were 96 ± 2 and 86.5 ± 7.2% before freezing, and 43 ± 5 and 67.4 ± 3.9% after thawing, respectively. Although the number of progressively motile sperm after thawing varied among samples (1.8 ± 1.2 × 10 8 cells/ejaculate), frozen semen in the present study might serve for artificial insemination. KEY WORDS: brown bear, freezing, spermatozoa.J. Vet. Med. Sci. 64(4): 373-376, 2002 The Hokkaido brown bears are found only in Hokkaido Island, Japan, and their genetic diversity is known to be low [15]. An assisted reproductive technology like artificial insemination could contribute to the maintainance of their genetic diversity. We previously reported a successful semen collection of the Hokkaido brown bear by electroejaculation [7]. Semen cryopreservation is an essential procedure for assisted breeding; however, there are only a few reports on cryopreservation of the bear semen [2,9]. The objective of the present study was to examine the potential of a freezing procedure used for bull semen with a modification in glycerol concentration.Four adult males, aged 7 to 16 years and maintained at Noboribetsu Bear Park (Noboribetsu, Japan), were used as semen donors. They were kept in male groups and separated from females. A total of 10 trials of electroejaculation were conducted during the breeding season (May to July) in 1996 and 1997. Electroejaculation was conducted as described previously [7]. Briefly, bears were immobilized with tiletamine HClzolazepam HCl (Zoletil; Virbac S. A., Carros, France) in 8 trials, and with midazolam (Dormicum; F. Hoffman-La Roche Ltd., Basel, Switzerland) and ketamine HCl (Ketalar; Sankyo Co., Ltd., Tokyo, Japan) in 2 trials. After immobilization, a handmade bipolar rectal probe with two ring electrodes was inserted into the rectum. The electrical stimulation with 60 Hz alternative current, sine wave and 5 V was repeated till ejaculates were obtained. The semen was assessed by determining ejaculate volume, pH, sperm concentration and the percentage of abnormal sperm as described elsewhere [7]. The percentage of viable spermatozoa was determined by an eosin-nigrosin staining method [7] before and after freezing. Small aliquot of raw and frozen-thawed semen was taken for the assessment of sperm motility. Percentage of sperm in each type of progressive motility status was calculated under a light microscope using a scale of 0 to 4: 0, no movement; 1, slight side-to-side movement with no forward progression; 2, side-to-side movement, slow and steady forward progression; 3, steady, moderate forward progression; 4, steady, rapid forward progression. The total perce...

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“…In recent years, their habitats have become fragmented and shrunken by human activity, and this species even faces extinction in some areas [7]. What is worse, bears have been killed to protect forestry, agriculture, livestock and humans from damage caused by them.Endangered and threatened wild animals are now attracting attention as a genetic resource and for species conservation, and semen has been recovered from various captive non-domestic animals, including fat-tailed dunnarts, koalas, brushtail possums, long-footed potoroos, northern brown bandicoots and ring-tailed possums [32] [4,20,22,24], Hokkaido brown bears [13,14] and Japanese black bears [16]. However, these studies were done with captive animals, and there are no reports on semen collection and cryopreservation in free-ranging bears.…”

mentioning

confidence: 99%

“…As to Ursidae, there are reports on electroejaculation and/or semen cryopreservation in giant pandas [4,20,22,24], Hokkaido brown bears [13,14] and Japanese black bears [16]. However, these studies were done with captive animals, and there are no reports on semen collection and cryopreservation in free-ranging bears.…”

mentioning

confidence: 99%

Electroejaculation and Semen Cryopreservation of Free-Ranging Japanese Black Bears (Ursus thibetanus japonicus)

Okano

Murase

Tsubota

2004

The Journal of Veterinary Medical Science

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ABSTRACT. The Japanese black bear (Ursus thibetanus japonicus) is endangered for extinction in some areas of Japan, and semen collection and cryopreservation are an important means to preserve genetic resources. The aim of this study was to characterize and c ryopreserve semen of free-ranging Japanese black bears. Semen was collected by electroejaculation procedure from 4 free-ranging Japanese black bears at the capture point in the field. Ejaculates containing motile sperm were recovered from all of the animals and ejaculate volume, total sperm count, % motility (percentage of motile spermatozoa), % viability (percentage of spermatozoa that excluded eosin) and % abnormal morphology (range (mean)) were 0.65-2.20 (1.51) ml, 99-1082 (490) × 10 6 , 5-100 (31), 42-97 (66) and 20-87 (53), respectively. Three of the 4 ejaculates were diluted with an egg yolk-TRIS-citrate-glucose extender and cryopreserved in liquid nitrogen. Motile spermatozoa were observed after freezing and thawing in all cases. This study showed that electroejaculation would be a useful method for collecting semen from free-ranging Japanese black bears and that at least motile spermatozoa would be obtained by fre ezing the thus collected electroejaculates. KEY WORDS: cryopreservation, electroejaculation, free-ranging, Japanese black bear, semen.J. Vet. Med. Sci. 66(11): 1371-1376, 2004 The Japanese black bear (Ursus thibetanus japonicus) is a large mammal that inhabits the islands of Honshu and Shikoku in Japan. In recent years, their habitats have become fragmented and shrunken by human activity, and this species even faces extinction in some areas [7]. What is worse, bears have been killed to protect forestry, agriculture, livestock and humans from damage caused by them.Endangered and threatened wild animals are now attracting attention as a genetic resource and for species conservation, and semen has been recovered from various captive non-domestic animals, including fat-tailed dunnarts, koalas, brushtail possums, long-footed potoroos, northern brown bandicoots and ring-tailed possums [32] [4,20,22,24], Hokkaido brown bears [13,14] and Japanese black bears [16]. However, these studies were done with captive animals, and there are no reports on semen collection and cryopreservation in free-ranging bears. Cryopreservation of semen collected from free-ranging animals is one of the most important methods for preservation of genetic resources and useful for artificial breeding. Moreover, characteristics of semen from wild animals may be essential for understanding reproductive status under field conditions, although studies using captive animals, kept under controlled conditions, may provide valuable data to analogize state under field conditions. It is easy to expect that the state of captive animals may differ from that of free-ranging ones to some extent, because of differences in their living environment and diet. It is difficult to obtain semen from free-ranging Japanese black bears due to limited access to the animals and seasonality of their ...

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Electroejaculation and semen analysis and freezing in the giant panda (Ailuropoda melanoleuca)

Cited by 44 publications

References 6 publications

Acrosomal integrity and capacitation are not influenced by sperm cryopreservation in the giant panda

Acrosomal integrity and capacitation are not influenced by sperm cryopreservation in the giant panda

Cryopreservation of the Semen Collected by Electroejaculation from the Hokkaido Brown Bear(Ursus arctos yesoensis).

Electroejaculation and Semen Cryopreservation of Free-Ranging Japanese Black Bears (Ursus thibetanus japonicus)

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