Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

Characteristics of high-quality Asian elephant (Elephas maximus) ejaculates and in vitro sperm quality after prolonged chilled storage and directional freezing

J. K. O’Brien A E , K. J. Steinman A , G. A. Montano A B , C. C. Love C , R. L. Saiers D and T. R. Robeck A
+ Author Affiliations
- Author Affiliations

A SeaWorld and Busch Gardens Reproductive Research Center, SeaWorld Parks and Entertainment, 2595 Ingraham St, San Diego, CA 92109, USA.

B Department of Animal Science, Texas A&M University, College Station, TX 77843, USA.

C Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX 77843, USA.

D Albuquerque Biological Park, Albuquerque, NM 87102, USA.

E Corresponding author. Email: justine.obrien@seaworld.com

Reproduction, Fertility and Development 25(5) 790-797 https://doi.org/10.1071/RD12129
Submitted: 21 February 2012  Accepted: 2 July 2012   Published: 15 August 2012

Abstract

The in vitro quality of spermatozoa from one elephant (Elephas maximus) was examined after chilled storage and directional freezing (DF). High-quality, non-contaminated ejaculates (77.6 ± 6.0% progressive motility, 3.9 ± 1.5 µg creatinine mL–1 raw semen, 2.7 ± 0.6% detached heads) were cryopreserved after 0 (0hStor), 12 (12hStor) and 24 h (24hStor) of chilled storage. At 0 h and 6 h post-thawing, total motility, plasma membrane integrity, acrosome integrity, mitochondrial activity and normal morphology were similar (P > 0.05) across treatments. In contrast, progressive motility, rapid velocity and several kinematic parameters were lower (P < 0.05) for 24Stor compared with 0hStor at 0 h post-thaw. By 6 h post-thaw, amplitude of lateral head displacement and velocity parameters (average pathway, straight-line and curvilinear velocity) were lower (P < 0.05) for 24hStor compared with 0hStor and 12hStor. DNA integrity was high and remained unchanged (P > 0.05) across all groups and processing stages (1.6 ± 0.6% of cells contained fragmented DNA). Results indicate that DF after up to 12 h of chilled storage results in a post-thaw sperm population of acceptable quality for artificial insemination. These findings have implications for the cryopreservation of sex-sorted spermatozoa, which typically undergo more than 12 h of chilled storage prior to sorting and preservation.

Additional keywords: CASA, DNA integrity, mitochondrial activity, SCSA, sperm cryopreservation.


References

Broekhuijse, M. L., Soštarić, E., Feitsma, H., and Gadella, B. M. (2011). Additional value of computer-assisted semen analysis (CASA) compared to conventional motility assessments in pig artificial insemination. Theriogenology 76, 1473–1486.
Additional value of computer-assisted semen analysis (CASA) compared to conventional motility assessments in pig artificial insemination.CrossRef | 1:STN:280:DC%2BC3Mfpt1Whuw%3D%3D&md5=6249df24533cc5402d90475bc884c2a8CAS | 21872316PubMed |

Didion, B. A., Kasperson, K. M., Wixon, R. L., and Evenson, D. P. (2009). Boar fertility and sperm chromatin structure status: a retrospective report. J. Androl. 30, 655–660.
Boar fertility and sperm chromatin structure status: a retrospective report.CrossRef | 19478334PubMed |

Evenson, D. P., Larson, K. L., and Jost, L. K. (2002). Sperm chromatin structure assay: its clinical use for detecting sperm DNA fragmentation in male infertility and comparisons with other techniques. J. Androl. 23, 25–43.
| 11780920PubMed |

Faust, L. J., Thompson, S. D., and Earnhardt, J. M. (2006). Is reversing the decline of Asian elephants in North American zoos possible? An individual-based modelling approach. Zoo Biol. 25, 201–218.
Is reversing the decline of Asian elephants in North American zoos possible? An individual-based modelling approach.CrossRef |

Gillan, L., Kroetsch, T., Maxwell, W. M. C., and Evans, G. (2008). Assessment of in vitro sperm characteristics in relation to fertility in dairy bulls. Anim. Reprod. Sci. 103, 201–214.
Assessment of in vitro sperm characteristics in relation to fertility in dairy bulls.CrossRef | 17208395PubMed |

Graham, E. F., Crabo, B. G., and Brown, K. (1972). I. Effect of some zwitterion buffers on the freezing and storage of spermatozoa I. Bull. J. Dairy Sci. 55, 372–378.
I. Effect of some zwitterion buffers on the freezing and storage of spermatozoa I. Bull.CrossRef | 1:CAS:528:DyaE38XhtFWju7o%3D&md5=9837c45c7476c17f28311e7139593141CAS | 5010387PubMed |

Gravance, C. G., Garner, D. L., Baumber, J., and Ball, B. A. (2000). Assessment of equine sperm mitochondrial function using JC-1. Theriogenology 53, 1691–1703.
Assessment of equine sperm mitochondrial function using JC-1.CrossRef | 1:STN:280:DC%2BD3cvpsFensg%3D%3D&md5=dcc9e747927b4a4a1baa4c3f4513dfedCAS | 10968415PubMed |

Hildebrandt, T. B., Hermes, R., Pratt, N. C., Fritsch, G., Blottner, S., Schmitt, D. L., Ratanakorn, P., Brown, J. L., Rietschel, W., and Göritz, F. (2000). Ultrasonography of the urogenital tract in elephants (Loxodonta africana and Elephas maximus): an important tool for assessing male reproductive function. Zoo Biol. 19, 333–345.
Ultrasonography of the urogenital tract in elephants (Loxodonta africana and Elephas maximus): an important tool for assessing male reproductive function.CrossRef |

Holt, C., Holt, W., Moore, H. D. M., Reed, H. C. B., and Curnock, R. M. (1997). Objectively measured boar sperm motility parameters correlate with the outcomes of on-farm inseminations: results of two fertility trials. J. Androl. 18, 312–323.
| 1:STN:280:DyaK2szks1Kntg%3D%3D&md5=d8f05bcd6467a1ee4f2897ecf5b20cf7CAS | 9203061PubMed |

Holt, W. V., O’Brien, J. K., and Abaigar, T. (2007). Applications and interpretation of computer-assisted sperm analyses and sperm-sorting methods in assisted breeding and comparative research. Reprod. Fertil. Dev. 19, 709–718.
Applications and interpretation of computer-assisted sperm analyses and sperm-sorting methods in assisted breeding and comparative research.CrossRef | 17714625PubMed |

Imrat, P., Hernandez, M., Rittem, S., Thongtip, N., Mahasawangkul, S., Gosálvez, J., and Holt, W. V. (2012). The dynamics of sperm DNA stability in Asian elephant (Elephas maximus) spermatozoa before and after cryopreservation. Theriogenology 77, 998–1007.
The dynamics of sperm DNA stability in Asian elephant (Elephas maximus) spermatozoa before and after cryopreservation.CrossRef | 1:CAS:528:DC%2BC38XivFOmtLY%3D&md5=4ab30337823ec12de41abaaf29b269e0CAS | 22225679PubMed |

IUCN (2010). The red list of threatened animals, In ‘IUCN Red List of Threatened Species’. Version 2009.2, (2010). Available from www.iucn.redlist.org [Verified 25 July 2012]

Kiso, W. K., Brown, J. L., Siewerdt, F., Schmitt, D. L., Olson, D., Crichton, E. G., and Pukazhenthi, B. S. (2011). Liquid semen storage in elephants (Elephas maximus and Loxodonta africana): species differences and storage optimisation. J. Androl. 32, 420–431.
Liquid semen storage in elephants (Elephas maximus and Loxodonta africana): species differences and storage optimisation.CrossRef | 21127305PubMed |

Love, C. C. (2011). Relationship between sperm motility, morphology and the fertility of stallions. Theriogenology 76, 547–557.
Relationship between sperm motility, morphology and the fertility of stallions.CrossRef | 1:STN:280:DC%2BC3MnkvF2jsw%3D%3D&md5=714d2150fc3ff3a6bffbddc43e591d99CAS | 21497893PubMed |

Marchetti, C., Jouy, N., Leroy-Martin, B., Defossez, A., Formstecher, P., and Marchetti, P. (2004). Comparison of four fluorochromes for the detection of the inner mitochondrial membrane potential in human spermatozoa and their correlation with sperm motility. Hum. Reprod. 19, 2267–2276.
Comparison of four fluorochromes for the detection of the inner mitochondrial membrane potential in human spermatozoa and their correlation with sperm motility.CrossRef | 15256505PubMed |

Marshburn, P. B., McIntire, D., Carr, B. R., and Byrd, W. (1992). Spermatozoal characteristics from fresh and frozen donor semen and their correlation with fertility outcome after intrauterine insemination. Fertil. Steril. 58, 179–186.
| 1:STN:280:DyaK38zisVequg%3D%3D&md5=f606251234360a03670f2740f000bbc9CAS | 1624002PubMed |

Montano, G. A., Kraemer, D. C., Love, C. C., Robeck, T. R., and O’Brien, J. K. (2012). Evaluation of motility, membrane status and DNA integrity of frozen–thawed bottlenose dolphin (Tursiops truncatus) spermatozoa after sex-sorting and recryopreservation. Reproduction 143, 799–813.
Evaluation of motility, membrane status and DNA integrity of frozen–thawed bottlenose dolphin (Tursiops truncatus) spermatozoa after sex-sorting and recryopreservation.CrossRef | 1:CAS:528:DC%2BC38XovFahs7k%3D&md5=cb6c0e1e19a63b8e1a63b6729112f5ccCAS | 22454530PubMed |

O’Brien, J. K., and Robeck, T. R. (2006). Development of sperm sexing and associated assisted reproductive technology for sex pre-selection of captive bottlenose dolphins (Tursiops truncatus). Reprod. Fertil. Dev. 18, 319–339.
Development of sperm sexing and associated assisted reproductive technology for sex pre-selection of captive bottlenose dolphins (Tursiops truncatus).CrossRef | 1:STN:280:DC%2BD287ms1Sltw%3D%3D&md5=fe479e3b19ad24233a0ce3cae715926bCAS | 16554007PubMed |

O’Brien, J. K., Steinman, K. J., and Robeck, T. R. (2009). Application of sperm sorting and associated reproductive technology for wildlife management and conservation. Theriogenology 71, 98–107.
Application of sperm sorting and associated reproductive technology for wildlife management and conservation.CrossRef | 1:STN:280:DC%2BD1cjnt1Khsg%3D%3D&md5=089fc8520e8489de31aa1bf5c6c33c6cCAS | 19010523PubMed |

O’Brien, J. K., Roth, T. L., Stoops, M. A., Ball, R. L., Steinman, K. J., Buescher, M. Y., Trujillo, T. P., Dragoo, G. A., Eisele, S. W., Montano, G. A., Wolfe, B. A., and Robeck, T. R. (2011). Sperm sorting and preservation technologies for sex ratio modification in the elephant and rhinoceros: an update. In ‘Proceedings of the 2011 International Elephant and Rhinoceros Conservation and Research Symposium, Rotterdam, The Netherlands’. pp. 39–40. [Abstract]

Robeck, T. R., Gearhart, S. A., Steinman, K. J., Katsumata, E., Loureiro, J. D., and O’Brien, J. K. (2011). In vitro sperm characterisation and development of a sperm cryopreservation method using directional solidification in the killer whale (Orcinus orca). Theriogenology 76, 267–279.
In vitro sperm characterisation and development of a sperm cryopreservation method using directional solidification in the killer whale (Orcinus orca).CrossRef | 1:STN:280:DC%2BC3MngtVSjug%3D%3D&md5=5db858f229d021fcd170d5b4ba53c142CAS | 21496896PubMed |

Sa-Ardrit, M., Saikhun, J., Thongtip, N., Damyang, M., Mahasawangkul, S., Angkawanish, T., Jansittiwate, S., Faisaikarm, T., Kitiyanant, Y., Pavasuthipaisit, K., and Pinyopummin, A. (2006). Ultrastructural alterations of frozen–thawed Asian elephant (Elephas maximus) spermatozoa. Int. J. Androl. 29, 346–352.
Ultrastructural alterations of frozen–thawed Asian elephant (Elephas maximus) spermatozoa.CrossRef | 1:STN:280:DC%2BD287ksFGksg%3D%3D&md5=fb31c66e5fe53da2031593f77bf5b8e2CAS | 16533357PubMed |

Saragusty, J., Hermes, R., Göritz, F., Schmitt, D. L., and Hildebrandt, T. B. (2009a). Skewed birth sex ratio and premature mortality in elephants. Anim. Reprod. Sci. 115, 247–254.
Skewed birth sex ratio and premature mortality in elephants.CrossRef | 19058933PubMed |

Saragusty, J., Hildebrandt, T. B., Behr, B., Knieriem, A., Kruse, J., and Hermes, R. (2009b). Successful cryopreservation of Asian elephant (Elephas maximus) spermatozoa. Anim. Reprod. Sci. 115, 255–266.
Successful cryopreservation of Asian elephant (Elephas maximus) spermatozoa.CrossRef | 1:CAS:528:DC%2BD1MXptl2iu78%3D&md5=8645f5e0665d2d6afd1cc2f0d52ce3e8CAS | 19111407PubMed |

Schmitt, D. L., and Hildebrandt, T. B. (1998). Manual collection and characterisation of semen from Asian elephants (Elephas maximus). Anim. Reprod. Sci. 53, 309–314.
Manual collection and characterisation of semen from Asian elephants (Elephas maximus).CrossRef | 1:STN:280:DyaK1M%2FltFSnug%3D%3D&md5=ede37b2baf7df4b598f7686cc703a829CAS | 9835384PubMed |

Taussky, H. H. (1954). A microcolorimetric determination of creatine in urine by the Jaffe reaction. J. Biol. Chem. 208, 853–861.
| 1:CAS:528:DyaG2cXlslSksQ%3D%3D&md5=c0c844fe595305d1b3f4fb0336a528a9CAS | 13174594PubMed |

Thongtip, N., Saikhun, J., Damyang, M., Mahasawangkul, S., Suthunmapinata, P., Yindee, M., Kongsila, A., Angkawanish, T., Jansittiwate, S., Wongkalasin, W., Wajjwalkul, W., Kitiyanant, Y., Pavasuthipaisit, K., and Pinyopummin, A. (2004). Evaluation of post-thaw Asian elephant (Elephas maximus) spermatozoa using flow cytometry: the effects of extender and cryoprotectant. Theriogenology 62, 748–760.
Evaluation of post-thaw Asian elephant (Elephas maximus) spermatozoa using flow cytometry: the effects of extender and cryoprotectant.CrossRef | 1:CAS:528:DC%2BD2cXltF2rs70%3D&md5=725fbe67d2ba7a55c9038e89ad5884feCAS | 15226027PubMed |

Thongtip, N., Saikhun, J., Mahasawangkul, S., Kornkaewrat, K., Pongsopavijitr, P., Songsasen, N., and Pinyopummin, A. (2008). Potential factors affecting semen quality in the Asian elephant (Elephas maximus). Reprod. Biol. Endocrinol. 6, 9.
Potential factors affecting semen quality in the Asian elephant (Elephas maximus).CrossRef | 18346275PubMed |

Thongtip, N., Mahasawangkul, S., Thitaram, C., Pongsopavijitr, P., Kornkaewrat, K., Pinyopummin, A., Angkawanish, T., Jansittiwate, S., Rungsri, R., Boonprasert, K., Wongkalasin, W., Homkong, P., Dejchaisri, S., Wajjwalku, W., and Saikhun, K. (2009). Successful artificial insemination in the Asian elephant (Elephas maximus) using chilled and frozen–thawed semen. Reprod. Biol. Endocrinol. 7, 75.
Successful artificial insemination in the Asian elephant (Elephas maximus) using chilled and frozen–thawed semen.CrossRef | 19615097PubMed |

Turner, R. M., Love, C. C., McDonnell, S. M., Sweeney, R. W., Twitchell, E. D., Habecker, P. L., Reilly, L. K., Pozor, M. A., and Kenney, R. M. (1995). Use of imipramine hydrochloride for treatment of urospermia in a stallion with a dysfunctional bladder. J. Am. Vet. Med. Assoc. 207, 1602–1606.
| 1:CAS:528:DyaK28XksFaqsg%3D%3D&md5=4906a9bfd7edfcdd7abe0f9fc07a49a4CAS | 7493900PubMed |



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