Evaluation of post-thaw Asian elephant (Elephas maximus) spermatozoa using flow cytometry: the effects of extender and cryoprotectant

Thongtip, Nikorn; Saikhun, Jumnian; Damyang, M.; Mahasawangkul, Sittidet; Suthunmapinata, P.; Yindee, Manoch; Kongsila, Apisek; Angkawanish, Tawepoke; Jansittiwate, Sarun; Wongkalasin, Waroot; Wajjwalkul, Worawidh; Kitiyanant, Yindee; Pavasuthipaisit, K.; Pinyopummin, Anuchai

doi:10.1016/j.theriogenology.2003.11.021

Cited by 34 publications

(34 citation statements)

References 27 publications

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“…Two extenders were selected for this study; INRA96 ® , a skimmed milk based extender (IMV Technologies, L'Aigle, France), and TEST an egg yolk based extender (Thongtip et al, 2004). INRA96…”

Section: Extenders and Cryoprotectantsmentioning

confidence: 99%

“…Freezing and thawing was performed as described previously by Thongtip et al (2004). In short, fresh semen was diluted with TEST or INRA96 ® to a concentration of 100 x 106 spermatozoa per ml immediately after collection.…”

Section: Semen Freezing and Thawingmentioning

confidence: 99%

“…The remainder of the diluted part of the sample was incubated in a water bath at 37 ºC and sub-samples were taken for DNA fragmentation analysis after incubation for 0, 4, 8, 24 and 48 h. DNA fragmentation (SDF%) was analyzed using the SPERM-HALOMAX ® kit (Chromacell SL, Madrid, Spain) for sperm chromatin dispersion as described previously (Imrat et al, 2012a); the time points were chosen specifically to examine the dynamics of DNA fragmentation as an additional 'stress test' for sperm quality. Sperm viability and acrosome integrity were assessed using a modification of the flow cytometric technique described by Thongtip et al (2004). Briefly, 1 ml of the undiluted thawed sample was mixed with 1 ml of Hepes-buffered modified Tyrode's medium (SP-TALP; 116 mM NaCl, 20 mM Hepes (pH 7.58), 5.0 mM Glucose, 3.1 mM KCl, 0.4 mM MgSO4•7H2O, 21.7 mM lactate, 0.3 mM NaH2PO4•H2O, 1 mM sodium pyruvate, 2.0 mM CaCl2, and 3mg/ml BSA; pH 7.4; osmolarity of 300 mOsm/kg) and centrifuged at 500 g for 10 min at room temperature.…”

Section: Post-thaw Evaluationmentioning

confidence: 99%

“…In addition, African elephant semen has been cryopreserved successfully, and this encouraged attempts to cryopreserve Asian elephant semen using the same extenders. Unfortunately, the post-thaw quality of cryopreserved Asian elephant semen proved to be very low, and relatively few bulls were capable of providing semen able to adequately survive the freezing process (Graham et al, 2004;Thongtip et al, 2004;Sa-Ardrit et al, 2006;Saragusty et al, 2009). While post-thaw semen quality depends on a variety of factors, fresh semen quality is used as one of the primary indicators of suitability for semen cryopreservation in many species; in particular, the percentage of motile sperm is a widely used criterion for selecting semen samples prior to freezing (Isachenko et al, 2004;Saragusty et al, 2009;Akhter et al, 2010;Stoops et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

“…In general, the components of a sperm cryopreservation extender include a non-permeating cryoprotectant, a penetrating cryoprotectant, a buffer, one or more sugars, salts and antibiotics (Evans and Maxwell, 1987). The earliest reports on the cryopreservation of Asian elephant sperm described the use of egg-yolk as the nonpermeating cryoprotectant and glycerol as the penetrating cryoprotectant (Thongtip et al, 2004;Saragusty et al, 2009). Recent studies have indicated that skim-milk powder is an equally effective non-permeating cryoprotectant for Asian elephant sperm (Saragusty et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

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Effect of pre-freeze semen quality, extender and cryoprotectant on the post-thaw quality of Asian elephant (Elephas maximus indicus) semen

et al. 2013

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Semen cryopreservation and artificial insemination (AI) are potentially valuable methods for supporting the breeding management of endangered species like the Asian elephant. Cryopreservation of Asian elephant semen has however proven problematic with respect to maintenance of both adequate semen quality and fertility post-thaw. In this study, nine ejaculates from three adult bulls were used to compare the influence of extender (TEST versus INRA96 ® ) and penetrating cryoprotectants (3% glycerol, 5% glycerol and 4% methylformamide) on postthaw semen quality. We demonstrate that not only the freezing process, but also the quality of the semen before freezing, significantly influences the freezability of Asian elephant semen. Prefreeze motility, viability, semen volume, semen pH, sperm concentration and the incidence of sperm mid-piece and tail abnormalities all significantly (p<0.05) affected post-thaw semen quality. While extender and cryoprotectant did not significantly affect any of the above semen quality parameters post-thaw, the skim-milk based extender (INRA96 ® ) preserved DNA integrity better (p<0.05) than the egg yolk extender (TEST). Considerable between-ejaculate variation in all postthaw semen quality parameters was also noted. It is concluded that strict criteria for semen quality is essential for the selection of Asian elephant bull ejaculates suitable for cryopreservation; stricter initial selection should improve the mean post-thaw quality.

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Section: Extenders and Cryoprotectantsmentioning

confidence: 99%

Section: Semen Freezing and Thawingmentioning

confidence: 99%

Section: Post-thaw Evaluationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effect of pre-freeze semen quality, extender and cryoprotectant on the post-thaw quality of Asian elephant (Elephas maximus indicus) semen

et al. 2013

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Reproductive System

Schmitt¹

2006

Biology, Medicine, and Surgery of Elephants

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Comparative Reproductive Biology of Elephants

Brown

2014

Advances in Experimental Medicine and Biology

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The ability to serially collect blood samples and conduct ultrasound examinations in Asian and African elephants has provided unique opportunities to study the biology of these endangered species. As a result, many unique aspects of elephant reproduction have been identified. For females, there are interesting differences in luteal steroidogenic activity, follicular maturation, pituitary gonadotropin secretion, fetal development and reproductive tract anatomy, while males exhibit the unique phenomenon of musth and an unusual reproductive anatomy (internal testes, ampullary semen storage). However, problems associated with uterine and ovarian pathologies hamper captive propagation efforts. Older, nulliparous cows are particularly susceptible, leading to speculation that continuous ovarian cyclicity of non-bred females in zoos is having a negative and cumulative effect on reproductive health. There are notable species differences in reproductive mechanisms as well (e.g., ovarian acyclicity, prolactin secretion, sperm cryosensitivity), implying that species-specific approaches to management and application of assisted reproductive techniques are needed for maximal reproductive efficiency and enhancement of genetic management.

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Evaluation of post-thaw Asian elephant (Elephas maximus) spermatozoa using flow cytometry: the effects of extender and cryoprotectant

Cited by 34 publications

References 27 publications

Effect of pre-freeze semen quality, extender and cryoprotectant on the post-thaw quality of Asian elephant (Elephas maximus indicus) semen

Effect of pre-freeze semen quality, extender and cryoprotectant on the post-thaw quality of Asian elephant (Elephas maximus indicus) semen

Reproductive System

Comparative Reproductive Biology of Elephants

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