European Journal of Wildlife Research

, Volume 53, Issue 2, pp 81–89 | Cite as

Semen cryopreservation and the conservation of endangered species

  • Jörns Fickel
  • Asja Wagener
  • Arne LudwigEmail author


Currently, more than 16,000 plant and animal species are imminently threatened by extinction, often as a direct consequence of anthropogenic influences. One of the measures to halt that process is genetic resource banking. This short review focuses on mammal sperm cryopreservation in combination with assisted reproduction techniques. It summarizes general problems, recent developments, and currently applied protocols and gives an overview of hitherto existing successes of assisted reproduction measures in wild animals in the light of conservation efforts.


Reproduction Spermatozoa 



artificial insemination


butylated hydroxytoluene




intracytoplasmic sperm injection


in vitro fertilization


seminal plasma



We value Steffen Blottner as a colleague and friend with whom we successfully cooperated during many years and whose enthusiastic discussions we always enjoyed, and we hope that we can further profit from his vast experience and knowledge in reproduction biology. Steffen Blottner, who together with other colleagues from the IZW initiated and maintained the institute’s spermbank, focused his research both on the establishment of applicable cryopreservation protocols for endangered species and on the exploration of pathways involved in and factors regulating spermatogenesis.


  1. Alm H, Torner H, Blottner S, Nürnberg G, Kanitz W (2001) Effect of sperm cryopreservation and treatment with calcium ionophore or heparin on in vitro fertilization of horse oocytes. Theriogenology 56:817–829PubMedCrossRefGoogle Scholar
  2. Almid T, Johnson L (1988) Effects of glycerol concentration, equilibration time and temperature of glycerol addition on post-thaw viability of boar spermatozoa frozen in straws. J Anim Sci 66:2899–2905Google Scholar
  3. Anel L, Martinez F, Alvarez M, Anel E, Boixo J, Kaabi M, Paz P, Chamorro C, Herraez P (1999) Post-mortem spermatozoa recovery and freezing in a Cantabrian brown bear (Ursus arctos): a preliminary report. Theriogenology 51:277CrossRefGoogle Scholar
  4. Asher GW, Kraemer DC, Magyar SJ, Brunner M, Moerbe R, Giaquinto M (1990) Intrauterine insemination of farmed fallow deer (Dama dama) with frozen–thawed semen via laparoscopy. Theriogenology 34:569–577PubMedCrossRefGoogle Scholar
  5. Asher GW, Fisher MW, Fennessy PF, Mackintosh CG, Jabbour HN, Morrow CJ (1993) Oestrous synchronization, semen collection and artificial insemination of farmed red deer (Cervus elaphus) and fallow deer (Dama dama). Anim Reprod Sci 33:241–265CrossRefGoogle Scholar
  6. Asher GW, Berg DK, Evans G (2000) Storage of semen and artificial insemination in deer. Anim Reprod Sci 62:195–211PubMedCrossRefGoogle Scholar
  7. Bains HK, Pabst MA, Werner G, Bawa SR (1993) Wheat-germ-agglutinin binding-sites on the sperm surface of water-buffalo (Bubalus bubalis)—a fracture label study. J Submicrosc Cytol Pathol 25:465–470PubMedGoogle Scholar
  8. Ballou JD (1992) Potential contribution of cryopreserved germ plasm to preservation of genetic diversity and conservation of endangered species in captivity. Cryobiology 29:19–25PubMedCrossRefGoogle Scholar
  9. Bartels P, Friedmann Y, Lubbe K, Mortimer D, Rasmussen LA, Godke RA (2001) The live birth of an eland (Taurotragus oryx) calf following estrous synchronization and artificial insemination using frozen thawed epididymal sperm. Theriogenology 55:381 (Abstract)Google Scholar
  10. Blottner S (1998) Semen preservation for the management of reproduction in rare and endangered species. Adv Ethol 33:9–13 (supplement to Ethology)Google Scholar
  11. Blottner S, Warnke C, Tuchscherer A, Heinen V, Torner H (2001) Morphological and functional changes of stallion spermatozoa after cryopreservation during breeding and non-breeding season. Anim Reprod Sci 65:75–88PubMedCrossRefGoogle Scholar
  12. Blottner S, Wagener A, Schön J, Göritz F, Fickel J (2006) Reproductive fitness in roe bucks (Capreolus capreolus): seasonal timing of testis function. Eur J Wildl Res 52:9–13CrossRefGoogle Scholar
  13. Byers AP, Hunter AG, Seal US, Binczik GA, Graham EF, Reindl NJ, Tilson RL (1989) In-vitro induction of capacitation of fresh and frozen spermatozoa of the Siberian tiger (Panthera tigris). J Reprod Fertil 86:599–607PubMedCrossRefGoogle Scholar
  14. Cardoso RDS, Silva AR, da Silva LDM (2006) Comparison of two dilution rates on canine semen quality after cryopreservation in a coconut water extender. Anim Reprod Sci 92:384–391CrossRefGoogle Scholar
  15. Choi YH, Love CC, Varner DD, Hinrichs K (2006) Equine blastocyst development after intracytoplasmic injection of sperm subjected to two freeze–thaw cycles. Theriogenology 65:808–819PubMedCrossRefGoogle Scholar
  16. Cirovic D (2006) First record of the racoon dog (Nyctereutes procyonoides Gray, 1834) in the former Yugoslav Republic of Macedonia. Eur J Wildl Res 52:136–137CrossRefGoogle Scholar
  17. Crump JP Jr, Crump JW (1994) Manual semen collection from a Grevy’s zebra stallion (Equus grevyi), onset of sperm production, semen characteristics, and cryopreservation of semen, with a comparison to the sperm production from a Grant’s Zebra stallion (Equus burchelli boehmi). Theriogenology 41:1011–1021PubMedCrossRefGoogle Scholar
  18. Cseh S, Solti L (2000) Importance of assisted reproductive technologies in the conservation of wild, rare or indigenous ungulates: a review article. Acta Vet Hung 48:313–323PubMedCrossRefGoogle Scholar
  19. Dankoff S, Curro T, Simmons H (2001) Post-thaw viability of wild cattle (Bos gaurus) and buffalo (Syncerus caffer) sperm cryopreserved using a novel, non-animal protein cryodiluent. Theriogenology 55:387 (Abstract)Google Scholar
  20. De Jong CE, Jonsson N, Field H, Smith C, Crichton EG, Phillips N, Johnston SD (2005) Collection, seminal characteristics and chilled storage of spermatozoa from three species of free-range flying fox (Pteropus spp.). Theriogenology 64:1072–1089PubMedCrossRefGoogle Scholar
  21. Drobnis EZ, Crowe LM, Berger T, Anchordoguy TJ, Overstreet JW, Crowe JH (1993) Cold shock damage is due to lipid phase-transitions in cell-membranes—a demonstration using sperm as a model. J Exp Zool 265:432–437PubMedCrossRefGoogle Scholar
  22. Durrant BS (1990) Semen collection, evaluation, and cryopreservation in exotic animal species: maximizing reproductive potential. ILAR News 32:2–10Google Scholar
  23. Farstad W, Fougner JA, Torres CG (1992) The optimum time for single artificial insemination of blue fox vixens (Alopex lagopus) with frozen–thawed semen from silver foxes (Vulpes vulpes). Theriogenology 38:853–865PubMedCrossRefGoogle Scholar
  24. Faszer K, Draper D, Green JE, Morris GJ, Grout BWW (2006) Cryopreservation of horse semen under laboratory and field conditions using a Stirling Cycle freezer. Cryo-lett 27:179–184Google Scholar
  25. Fickel J, Schmidt A, Putze M, Spittler H, Ludwig A, Streich WJ, Pitra C (2005) Genetic structure of populations of European brown hare: implications for management. J Wildl Manage 69:760–771CrossRefGoogle Scholar
  26. Freyhof J, Lieckfeldt D, Pitra C, Ludwig A (2005) Molecules and morphology: evidence for introgression of mitochondrial DNA in Dalmatian cyprinids. Mol Phylogenet Evol 37:347–354PubMedCrossRefGoogle Scholar
  27. Gao DY, Ashworth E, Watson PF, Kleinhans FW, Mazur P, Critser JK (1993) Hyperosmotic tolerance of human spermatozoa—separate effects of glycerol, sodium-chloride, and sucrose on spermolysis. Biol Reprod 49:112–123PubMedCrossRefGoogle Scholar
  28. Gao DY, Liu J, Liu C, McGann LE, Watson PF, Kleinhans FW, Mazur P, Critser ES, Critser JK (1995) Prevention of osmotic injury to human spermatozoa during addition and removal of glycerol. Hum Reprod 10:1109–1122PubMedGoogle Scholar
  29. Gilmore JA, McGann LE, Ashworth E, Acker JP, Raath JP, Bush M, Critser JK (1998) Fundamental cryobiologyogy of selected African mammalian spermatozoa and its role in biodiversity preservation through the development of genome resource banking. Anim Reprod Sci 53:277–297PubMedCrossRefGoogle Scholar
  30. Goodrowe KL, Hay MA, Platz CC, Behrns SK, Jones MH, Waddell WT (1998) Characteristics of fresh and frozen–thawed red wolf (Canis rufus) spermatozoa. Anim Reprod Sci 53:299–308PubMedCrossRefGoogle Scholar
  31. Haigh JC, Bowen G (1991) Artificial insemination of red deer (Cervus elephas) with frozen–thawed wapiti semen. J Reprod Fertil 93:119–123PubMedCrossRefGoogle Scholar
  32. Haigh JC, Dradjat AS, English AW (1993) Comparison of two extenders for the cryopreservation of chital (Axis axis) semen. J Zoo Wildl Med 24:454–458Google Scholar
  33. Hammadeh ME, Szarvasy D, Zeginiadou T, Rosenbaum P, Georg T, Schmidt W (2001) Evaluation of cryoinjury of spermatozoa after slow (programmed biological freezer) or rapid (liquid nitrogen vapour) freeze–thawing techniques. J Assist Reprod Genet 18:364–370PubMedCrossRefGoogle Scholar
  34. Hartley M (2005) A seroprevalence survey of Toxoplasma gondii in common wombats (Vombatus ursinus). Eur J Wildl Res 51:65–67CrossRefGoogle Scholar
  35. Hell P, Plavy R, Slamecka J, Gasparik J (2005) Losses of mammals (Mammalia) and birds (Aves) on roads in the Slovak part of the Danube Basin. Eur J Wildl Res 51:35–40CrossRefGoogle Scholar
  36. Hermes R, Goritz F, Maltzan J, Blottner S, Proudfoot J, Fritsch G, Fassbender M, Quest M, Hildebrandt TB (2001) Establishment of assisted reproduction technologies in female and male African wild dogs (Lycaon pictus). J Reprod Fertil Suppl 57:315–321PubMedGoogle Scholar
  37. Hermes R, Hildebrandt TB, Blottner S, Walzer C, Silinski S, Patton ML, Wibbelt G, Schwarzenberger F, Göritz F (2005) Reproductive soundness of captive southern and northern white rhinoceroses (Ceratotherium simum simum, C. s. cottoni): evaluation of male genital tract morphology and semen quality before and after cryopreservation. Theriogenology 63:219–238PubMedCrossRefGoogle Scholar
  38. Hildebrandt TB, Göritz F, Hermes R (2006) Ultrasonography: an important tool in captive breeding management in elephants and rhinoceroses. Eur J Wildl Res 52:23–27CrossRefGoogle Scholar
  39. Hishinuma M, Suzuki K, Sekine J (2003) Recovery and cryopreservation of sika deer (Cervus nippon) spermatozoa from epididymides stored at 4 degrees C. Theriogenology 59:813–820PubMedCrossRefGoogle Scholar
  40. Holt WV (1996) Can we predict fertility rates? Making sense of sperm motility. Reprod Domest Anim 31:17–24Google Scholar
  41. Holt WV (2000) Basic aspects of frozen storage of semen. Anim Reprod Sci 62:3–22PubMedCrossRefGoogle Scholar
  42. Holt WV, Pickard AR (1999). Role of reproductive technologies and genetic resource banks in animal conservation. Rev Reprod 4:143–150 (Review)PubMedCrossRefGoogle Scholar
  43. Holt WV, Bennett PM, Volobouev V, Watson PF (1996) Genetic resource banks in wildlife conservation. J Zool (London) 238:531–544Google Scholar
  44. Holt WV, Medrano A, Thurston LM, Watson PE (2005) The significance of cooling rates and animal variability for boar sperm cryopreservation: insights from the cryomicroscope. Theriogenology 63:370–382PubMedCrossRefGoogle Scholar
  45. Howard JG, Bush M, de Vos V, Schiewe MC, Pursel VG, Wildt DE (1986) Influence of cryoprotective diluent on post-thaw viability and acrosomal integrity of spermatozoa of the African elephant (Loxodonta africana). J Reprod Fertil 78:295–306PubMedCrossRefGoogle Scholar
  46. Howard JG, Bush M, Morton C, Morton F, Wentzel K, Wildt DE (1991) Comparative semen cryopreservation in ferrets (Mustela putorius furo) and pregnancies after laparoscopic intrauterine insemination with frozen–thawed spermatozoa. J Reprod Fertil 92:109–118PubMedCrossRefGoogle Scholar
  47. IUCN (2006) International Union for Conservation of Nature and Natural Resources, accessible at
  48. Jabbour HN, Argo CMcG, Brinklow BR, Loudon ASI, Hooton J (1993a). Concenption rates following intrauterine insemination of European fallow deer (Dama dama dama) does with fresh or frozen–thawed Mesopotamian fallow deer (Dama dama mesopotamica) spermatozoa. J Zool 230:379–384CrossRefGoogle Scholar
  49. Jabbour HN, Veldhuizen FA, Green G, Asher GW (1993b). Endocrine responses and conception rates in fallow deer (Dama dama) following oestrus synchronization and cervical insemination with fresh or frozen–thawed spermtozoa. J Reprod Fertil 98:495–502PubMedCrossRefGoogle Scholar
  50. Jenneckens I, Meyer JN, Debus L, Pitra C, Ludwig A (2000) Evidence of mitochondrial DNA clones of Siberian sturgeon, Acipenser baerii, within Russian sturgeon, Acipenser gueldenstaedtii, caught in the River Volga. Ecol Lett 3:503–508CrossRefGoogle Scholar
  51. Jetschke G (1992) Stochastic population-models and their relevance for the conservation of species. Ecol Model 63:71–89CrossRefGoogle Scholar
  52. Johnston SD, MacCallum C, Blyde D, McClean R, Lisle A, Holt WV (2006) An investigation into the similarities and differences governing the cryopreservation success of koala (Phascolarctos cinereus: goldfuss) and common wombat (Vombatus ursinus: shaw) spermatozoa. Cryobiology 53:218–228PubMedCrossRefGoogle Scholar
  53. Johnston SD, Ward D, Lemon J, Gunn I, Maccallum CA, Keeley T, Blyde D (2007) Studies of male reproduction in captive African wild dogs (Lycaon pictus). Anim Reprod Sci (in press) DOI  10.1016/j.anireprosci.2006.08.017
  54. Kierdorf U, Kierdorf H (2005) Antlers as biomonitors of environmental pollution by lead and fluoride: a review. Eur J Wildl Res 51:137–150CrossRefGoogle Scholar
  55. Kitiyanant Y, Schmidt MJ, Pavasuthipaisit K (2000) Evaluation of sperm acrosome reaction in the Asiatic elephant. Theriogenology 53:887–896PubMedCrossRefGoogle Scholar
  56. Kumar S, Millar JD, Watson PF (2003) The effect of cooling rate on the survival of cryopreserved bull, ram, and boar spermatozoa: a comparison of two controlled-rate cooling machines. Cryobiology 46:246–253PubMedCrossRefGoogle Scholar
  57. Leibo SP, Songsasen N (2002) Cryopreservation of gametes and embryos of non-domestic species. Theriogenology 57:303–326PubMedCrossRefGoogle Scholar
  58. Lieckfeldt D, Hett AK, Ludwig A, Freyhof J (2006) Detection, characterization and utility of a new highly variable nuclear marker region in several species of cyprinid fishes (Cyprinidae). Eur J Wildl Res 52:63–65CrossRefGoogle Scholar
  59. Loskutoff NM, Simmons HA, Goulding M, Thompson G, Jongh TD, Simmons LG (1996) Species and individual variations in cryoprotectant toxicities and freezing resistance of epididymal sperm from African antelope. Anim Reprod Sci 42:527–535CrossRefGoogle Scholar
  60. Ludwig A (2006) A sturgeon view on conservation genetics. Eur J Wildl Res 52:3–8CrossRefGoogle Scholar
  61. Ludwig A, Debus L, Lieckfeldt D, Wirgin I, Benecke N, Jenneckens I, Williot P, Waldman JR, Pitra C (2002a) When the American sea sturgeon swam east. Nature 419:447–448PubMedCrossRefGoogle Scholar
  62. Ludwig A, Debus L, Jenneckens I (2002b) A molecular approach for trading control of black caviar. Int Rev Hydrobiol 87:661–674CrossRefGoogle Scholar
  63. Ludwig A, Congiu L, Pitra C, Fickel J, Gessner J, Fontana F, Patarnello T, Zane L (2003) Nonconcordant evolutionary history of maternal and paternal lineages in Adriatic sturgeon. Mol Ecol 12:3253–3264PubMedCrossRefGoogle Scholar
  64. Luenser K, Fickel J, Lehnen A, Speck S, Ludwig A (2005) Low level of genetic variability in European bisons (Bison bonasus) from the Bialowieza National Park in Poland. Eur J Wildl Res 51:84–87CrossRefGoogle Scholar
  65. Lyons G (2006) Viewpoint: policy requirements for protecting wildlife from endocrine disruptors. Environ Health Perspect 114(suppl 1):142–146. Available at DOI  10.1289/ehp.8070 Google Scholar
  66. Martinez AIP (2004) Canine fresh and cryopreserved semen evaluation. Anim Reprod Sci 82–83:209–224PubMedCrossRefGoogle Scholar
  67. Martinez-Pastor F, Anel L, Guerra C, Alvarez M, Soler AJ, Garde JJ, Chamorro C, de Paz P (2006) Seminal plasma improves cryopreservation of Iberian red deer epididymal sperm. Theriogenology 66:1847–1856PubMedCrossRefGoogle Scholar
  68. McClean R, MacCallum C, Blyde D, Holt WV, Johnston SD (2006) Ultrastructure, osmotic tolerance, glycerol toxicity and cryopreservation of caput and cauda epididymal kangaroo spermatozoa. Reprod Fertil Dev 18:469–476PubMedCrossRefGoogle Scholar
  69. Miller RR Jr, Cornett CL, Waterhouse KE, Farstad W (2005) Comparative aspects of sperm membrane fatty acid composition in silver (Vulpes vulpes) and blue (Alopex lagopus) foxes and their relationship to cell cryopreservation. Cryobiology 51:66–75PubMedCrossRefGoogle Scholar
  70. Moce E, Graham JK, Schenk JL (2006) Effect of sex-sorting on the ability of fresh and cryopreserved bull sperm to undergo an acrosome reaction. Theriogenology 66:929–936PubMedCrossRefGoogle Scholar
  71. Molinia FC, Rodger JC (1996) Pellet-freezing spermatozoa of two marsupials: the tammar wallaby, Macropus eugenii, and the brushtail possum, Trichosurus vulpecula. Reprod Fertil Dev 8:681–684PubMedCrossRefGoogle Scholar
  72. Monfort SL, Asher GW, Wildt DE, Wood TC, Schiewe MC, Williamson LR, Bush M, Rall WF (1993) Successful intrauterine insemination of Eld’s deer (Cervus eldii thamin) with frozen–thawed spermatozoa. J Reprod Fertil 99:459–465PubMedCrossRefGoogle Scholar
  73. Morrell JM, Nubbemeyer R, Heistermann M, Rosenbusch J, Kuderling I, Holt W, Hodges JK (1998) Artificial insemination in Callithrix jacchus using fresh or cryopreserved sperm. Anim Reprod Sci 52:165–174PubMedCrossRefGoogle Scholar
  74. Mosaferi S, Niasari-Naslaji A, Abarghani A, Gharahdaahi AA, Gerami A (2005) Biophysical and biochemical characteristics of bactrian camel semen collected by artificial vagina. Theriogenology 63:92–101PubMedCrossRefGoogle Scholar
  75. Mulley RC, Moore NW, English AW (1988) Successful uterine insemination of fallow deer with fresh and frozen semen. Theriogenology 29:1149–1153PubMedCrossRefGoogle Scholar
  76. Neubauer K, Jewgenow K, Blottner S, Wildt DE, Pukazhenthi BS (2004) Quantity rather than quality in teratospermic males: a histomorphometric and flow cytometric evaluation of spermatogenesis in the domestic cat (Felis catus). Biol Reprod 71:1517–1524PubMedCrossRefGoogle Scholar
  77. Nichols SM, Bavister BD (2006) Comparison of protocols for cryopreservation of rhesus monkey spermatozoa by post-thaw motility recovery and hyperactivation. Reprod Fertil Dev 18:777–780PubMedCrossRefGoogle Scholar
  78. Nilsen EB, Solberg EJ (2006) Patterns of hunting mortality in Norwegian moose (Alces alces) populations. Eur J Wildl Res 52:153–163CrossRefGoogle Scholar
  79. O’Brien JK, Roth TL (2000a) Post-coital sperm recovery and cryopreservation in the Sumatran rhinoceros (Dicerorhinus sumatrensis) and application to gamete rescue in the African black rhinoceros (Diceros bicornis). J Reprod Fertil 118:263–271PubMedCrossRefGoogle Scholar
  80. O’Brien JK, Roth TL (2000b) Functional capacity and fertilizing longevity of frozen–thawed scimitar-horned oryx (Oryx dammah) spermatozoa in a heterologous in vitro fertilization system. Reprod Fertil Dev 12:413–421PubMedCrossRefGoogle Scholar
  81. Olson MA, Huang Y, Li DS, Spindler R, Howard J, Zhang HM, Durrant B (2003) Assessment of motility, acrosomal integrity, and viability of giant panda (Ailuropoda melanoleuca) sperm following short-term storage at 4 degrees C. Zoo Biol 22:529–544CrossRefGoogle Scholar
  82. Parks JE, Meacham TN, Saacke RG (1981) Cholesterol and phospholipids of bovine spermatozoa. 1. Selection of a PIPES-buffered diluent for evaluating the effect of egg-yolk lipoproteins on sperm cholesterol and phospholipids. Biol Reprod 24:393–398PubMedCrossRefGoogle Scholar
  83. Paz RCR, Zuge RM, Morato RG, Barnabe VC (2002) Penetration assay of frozen jaguar (Panthera onca) sperm in heterologous oocytes. Theriogenology 57:588 (Abstract)Google Scholar
  84. Persson Y, Soderquist L (2006) Comparison between spermiograms in semen samples obtained from bulls by transrectal massage and artificial vagina. Reprod Domest Anim 41:311 (Abstract)CrossRefGoogle Scholar
  85. Pickard AR, Abaigar T, Green DI, Holt WV, Cano M (2001) Hormonal characterization of the reproductive cycle and pregnancy in the female Mohor gazelle (Gazella dama mhorr). Reproduction 122:571–580PubMedCrossRefGoogle Scholar
  86. Pitra C, Lutz W (2005) Population genetic structure and the effect of founder events on the genetic variability of introduced sika deer, Cervus nippon, in Germany and Austria. Eur J Wildl Res 51:95–100CrossRefGoogle Scholar
  87. Pitra C, Rehbein S, Lutz W (2005) Tracing the genetic roots of the sika deer Cervus nippon naturalized in Germany and Austria. Eur J Wildl Res 51:237–241CrossRefGoogle Scholar
  88. Pitra C, VazPinto P, O’Keeffe BWJ, Willows-Munro S, van Vuuren BJ, Robinson TJ (2006) DNA-led rediscovery of the giant sable antelope in Angola. Eur J Wildl Res 52:142–152CrossRefGoogle Scholar
  89. Platz CC, Magyar S, Crider N, Densmore M, Wiley G, Browen MJ, Templeton JW, Kraemer DC (1982) Cryopreservation of electroejaculated and epididymal spermtozoa in white tail deer (Odocoileus virginiamus). In: Annual Proceedings—American Association of Zoo Veterinarians, pp 127–129Google Scholar
  90. Potts GR (2005) Incidence of ingested lead gunshot in wild grey partridges (Perdix perdix) from the UK. Eur J Wildl Res 51:31–34CrossRefGoogle Scholar
  91. Ptak G, Clinton M, Barboni B, Muzzeddu M, Cappai P, Tischner M, Loi P (2002) Preservation of the wild European mouflon: the first example of genetic management using a complete program of reproductive biotechnologies. Biol Reprod 66:796–801PubMedCrossRefGoogle Scholar
  92. Pukazhenthi B, Laroe D, Crosier A, Bush LM, Spindler R, Pelican KM, Bush M, Howard JG, Wildt DE (2006) Challenges in cryopreservation of clouded leopard (Neofelis nebulosa) spermatozoa. Theriogenology 66:1790–1796PubMedCrossRefGoogle Scholar
  93. Robeck TR, O’Brien JK (2004) Effect of cryopreservation methods and precryopreservation storage on bottlenose dolphin (Tursiops truncatus) spermatozoa. Biol Reprod 70:1340–1348PubMedCrossRefGoogle Scholar
  94. Roca J, Gil MA, Hernandez M, Parilla I, Vasquez JM, Martinez EA (2004) Survival and fertility of boar spermatozoa after freeze–thawing in extender supplemented with butylated hydroxytoluene. J Androl 25:397–405PubMedGoogle Scholar
  95. Sabocanec R, Konjevic D, Srebocan E, Petrinec Z (2005) Fatal poisoning of a Griffon vulture (Gyps fulvus) with methomyl. Eur J Wildl Res 51:210–212CrossRefGoogle Scholar
  96. Said S, Pellerin M, Guillon N, Debias F, Fritz H (2005) Assessment of forage availability in ecological studies. Eur J Wildl Res 51:242–247CrossRefGoogle Scholar
  97. Sanchez-Partida LG, Maginnis G, Dominko T, Martinovich C, McVay B, Fanton J, Schatten G (2000) Live rhesus offspring by artificial insemination using fresh sperm and cryopreserved sperm. Biol Reprod 63:1092–1097CrossRefGoogle Scholar
  98. Sankai T, Terao K, Yanagimachi R, Cho F, Yoshikawa Y (1994) Cryopreservation of spermtozoa from cynomolgus monkeys (Macaca fascicularis). J Reprod Fertil 101:273–278PubMedCrossRefGoogle Scholar
  99. Sanz-Elorza M, Dana ED, González A, Sobrino E (2003) Changes in the High-mountain vegetation of the Central Iberian Peninsula as a probable sign of global warming. Ann Bot (London) 92:273–280CrossRefGoogle Scholar
  100. Saragusty J, Hildebrandt TB, Natan Y, Hermes R, Yavin S, Göritz F, Arav A (2005) Effect of egg-phosphatidylcholine on the chilling sensitivity and lipid phase transition of Asian elephant (Elephas maximus) spermatozoa. Zoo Biol 24:233–245CrossRefGoogle Scholar
  101. Saragusty J, Gacitua H, King R, Arav A (2006) Post-mortem semen cryopreservation and characterization in two different endangered gazelle species (Gazella gazella and Gazella dorcas) and one subspecies (Gazella gazelle acaiae). Theriogenology 66:775–784PubMedCrossRefGoogle Scholar
  102. Seager SWJ, Platz CC, Hodge W (1975) Successful pregnancy using frozen semen in the wolf. Int Zoo Yearb 15:140–143Google Scholar
  103. Serrano E, Calabuig G, Perez JM, Gallego L (2005) The potential use of identification of skeletal remains for the early detection of Ammotragus: an exotic ungulate species in Southern Spain. Eur J Wildl Res 51:88–94CrossRefGoogle Scholar
  104. Si W, Hildebrandt TB, Reid C, Krieg R, Ji WZ, Fassbender M, Hermes R (2006) The successful double cryopreservation of rabbit (Oryctolagus cuniculus) semen in large volume using the directional freezing technique with reduced concentration of cryoprotectant. Theriogenology 65:788–798PubMedCrossRefGoogle Scholar
  105. Spindler RE, Yan H, Howard J, Pengyan W, Hemin Z, Guiquan Z, Wildt DE (2006) Giant panda (Ailuropoda melanoleuca) spermatozoa decondensation in vitro is not compromised by cryopreservation. Reprod Fertil Dev 18:767–775PubMedCrossRefGoogle Scholar
  106. Stilley K, Godke RA (1998) Assisted reproductive technologies in exotic zoo animals. Can Soc Anim Sci 98:213–239Google Scholar
  107. Swanson WF, Brown JL (2004) International training programs in reproductive sciences for conservation of Latin American felids. Anim Reprod Sci 82–83:21–34PubMedCrossRefGoogle Scholar
  108. Swanson WF, Howard JG, Roth TL, Brown JL, Alvarado T, Burton M, Starnes D, Wildt DE (1996a) Responsiveness of ovaries to exogenous gonadotrophins and laparoscopic artificial insemination with frozen–thawed spermatozoa in ocelots (Felis pardalis). J Reprod Fertil 106:87–94PubMedCrossRefGoogle Scholar
  109. Swanson WF, Roth TL, Blumer E, Citino SB, Kenny D, Wildt DE (1996b). Comparative cryopreservation and functionality of spermatozoa from the normospermic jaguar (Panthera onca) and teratospermic cheeta (Acinonyx jubatus). Theriogenology 45:241 (Abstract)CrossRefGoogle Scholar
  110. Swanson WF, Johnson WE, Cambre RC, Citino SB, Quigley KB, Brousset DM, Morais RN, Moreira N, O’Brien SJ, Wildt DE (2003) Reproductive status of endemic felid species in Latin America zoos and implications for ex situ conservation. Zoo Biol 22:421–441CrossRefGoogle Scholar
  111. Thiangtum K, Swanson WF, Howard J, Tunwattana W, Tongthainan D, Wichasilpa W, Patumrattanathan P, Pinyopoommintr T (2006) Assessment of basic seminal characteristics, sperm cryopreservation and heterologous in vitro fertilisation in the fishing cat (Prionailurus viverrinus). Reprod Fertil Dev 18:373–382PubMedCrossRefGoogle Scholar
  112. 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, Pinyopummin A (2004) Evaluation of post-thaw Asian elephant (Elephas maximus) spermatozoa using flow cytometry: the effects of extender and cryoprotectant. Theriogenology 62:748–760PubMedCrossRefGoogle Scholar
  113. Thurston LM, Watson PF, Holt WV (2002a) Semen cryopreservation: a genetic explanation for species and individual variation? Cryo-lett 23:255–262Google Scholar
  114. Thurston LM, Siggins K, Mileham AJ, Watson PF, Holt WV (2002b) Identification of amplified restriction fragment length polymorphism markers linked to genes controlling boar sperm viability following cryopreservation. Biol Reprod 66:545–554PubMedCrossRefGoogle Scholar
  115. Tollner TL, VandeVoort CA, Overstreet JW, Drobnis EZ (1990) Cryopreservation of spermtozoa from cynomoglus monkeys (Macaca fascicularis). J Reprod Fertil 90:347–352PubMedCrossRefGoogle Scholar
  116. Walters C, Wheeler L, Stanwood PC (2004) Longevity of cryogenically stored seeds. Cryobiology 48:229–244PubMedCrossRefGoogle Scholar
  117. Warnke C, Tuchscherer A, Alm H, Kanitz W, Blottner S, Torner H (2003) Characterisation of movement pattern and velocities of stallion spermatozoa depending on donor, season and cryopreservation. Acta Vet Hung 51:395–408PubMedCrossRefGoogle Scholar
  118. Wu JM, Chung YK, Belford KJ, Smith GD, Takayama S, Lahann J (2006) A surface-modified sperm sorting device with long-term stability. Biomed Microdevices 8:99–107PubMedCrossRefGoogle Scholar
  119. Zindl C, Asa CS, Gunzel-Apel AR (2006) Influence of cooling rates and addition of Equex pasta on cooled and frozen-thawed semen of generic gray (Canis lupus) and Mexican gray wolf (C. l. baileyi). Theriogenology 66:1797–1802PubMedCrossRefGoogle Scholar
  120. Zomborszky Z, Zubor T, Toth J, Horn P (1999) Sperm collection from shot red deer stags (Cervus elaphus) and the utilization of sperm frozen and subsequently thawed. Acta Vet Hung 47:263–270PubMedGoogle Scholar
  121. Zomborszky Z, Nagy S, Nanassy L, Szabari M, Bodo S (2005) Experiences in deer sperm cryopreservation under practical conditions—a pilot study. Anim Reprod Sci 90:185–190PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  1. 1.Department of Evolutionary GeneticsLeibniz Institute for Zoo and Wildlife ResearchBerlinGermany
  2. 2.Institute of Animal SciencesHumboldt-University BerlinBerlinGermany

Personalised recommendations