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Sperm cryopreservation and reproductive characteristics in the beauty snake (Elaphe taeniura) as a model for endangered Colubridae

Penelope Banchi (UGent) , Ferran Solanes Vilanova (UGent) , Mattia Cesauri, Lotte Spanoghe (UGent) , Koen Chiers (UGent) , Ann Van Soom (UGent) , Tom Hellebuyck (UGent) and Guilherme Rizzoto (UGent)
(2025) CRYOBIOLOGY. 118.
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Abstract
Sperm collection and cryopreservation are key technologies for assisted reproduction, with post-mortem sperm collection being the main tool to prevent the decline of endangered species. The present study describes postmortem sperm collection and two cryopreservation protocols in beauty snakes ( Elaphe taeniura), as a model for Colubridae. The vasa deferentia of 18 snakes were collected post-mortem and incubated for 30 min at room temperature to retrieve sperm by float up. Afterwards, fresh sperm was analysed, and 11 samples were diluted into either a single or a double-step TEST-egg yolk-based extender to reach a final concentration of 12 % glycerol. Anatomical and histological analyses of the reproductive organs revealed that those on the right side were larger, heavier, and positioned more cranially than the left ones (P = 0.007, P = 0.003, and P = 0.0002, respectively), but sperm parameters did not differ. Successful sperm retrieval was achieved in 61.1 % of snakes, with positive correlations between seminiferous tubules length and body weight (P = 0.0004) and between seminiferous tubules length and body length (P = 0.001). Sperm-producing animals were heavier and longer compared to azoospermic ones (P = 0.04 and P = 0.04 respectively). Specifically, males longer than 130 cm and heavier than 177g have higher reproductive potential. Both cryopreservation protocols yielded comparable results, preserving sperm viability, motility, and morphology. Therefore, a single-step protocol should be preferred due to its technical simplicity. In conclusion, we described for the first time the reproductive features and characteristics of spermatozoa collected post-mortem in beauty snakes. Although the described protocols are effective, further research is warranted to optimize these techniques.
Keywords
Snake, Sperm, Reproductive assisted technologies, Sperm freezing, Colubridae, Reptile, Semen analysis, 2-STEP DILUTION, STORAGE, SEMEN, CONSERVATION, SPERMATOZOA, PARAMETERS, SURVIVAL, EQUEX

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MLA
Banchi, Penelope, et al. “Sperm Cryopreservation and Reproductive Characteristics in the Beauty Snake (Elaphe Taeniura) as a Model for Endangered Colubridae.” CRYOBIOLOGY, vol. 118, 2025, doi:10.1016/j.cryobiol.2024.105184.
APA
Banchi, P., Solanes Vilanova, F., Cesauri, M., Spanoghe, L., Chiers, K., Van Soom, A., … Rizzoto, G. (2025). Sperm cryopreservation and reproductive characteristics in the beauty snake (Elaphe taeniura) as a model for endangered Colubridae. CRYOBIOLOGY, 118. https://doi.org/10.1016/j.cryobiol.2024.105184
Chicago author-date
Banchi, Penelope, Ferran Solanes Vilanova, Mattia Cesauri, Lotte Spanoghe, Koen Chiers, Ann Van Soom, Tom Hellebuyck, and Guilherme Rizzoto. 2025. “Sperm Cryopreservation and Reproductive Characteristics in the Beauty Snake (Elaphe Taeniura) as a Model for Endangered Colubridae.” CRYOBIOLOGY 118. https://doi.org/10.1016/j.cryobiol.2024.105184.
Chicago author-date (all authors)
Banchi, Penelope, Ferran Solanes Vilanova, Mattia Cesauri, Lotte Spanoghe, Koen Chiers, Ann Van Soom, Tom Hellebuyck, and Guilherme Rizzoto. 2025. “Sperm Cryopreservation and Reproductive Characteristics in the Beauty Snake (Elaphe Taeniura) as a Model for Endangered Colubridae.” CRYOBIOLOGY 118. doi:10.1016/j.cryobiol.2024.105184.
Vancouver
1.
Banchi P, Solanes Vilanova F, Cesauri M, Spanoghe L, Chiers K, Van Soom A, et al. Sperm cryopreservation and reproductive characteristics in the beauty snake (Elaphe taeniura) as a model for endangered Colubridae. CRYOBIOLOGY. 2025;118.
IEEE
[1]
P. Banchi et al., “Sperm cryopreservation and reproductive characteristics in the beauty snake (Elaphe taeniura) as a model for endangered Colubridae,” CRYOBIOLOGY, vol. 118, 2025.
@article{01JEB1Z6AEYV0CT51YSWEGMK7T,
  abstract     = {{Sperm collection and cryopreservation are key technologies for assisted reproduction, with post-mortem sperm collection being the main tool to prevent the decline of endangered species. The present study describes postmortem sperm collection and two cryopreservation protocols in beauty snakes ( Elaphe taeniura), as a model for Colubridae. The vasa deferentia of 18 snakes were collected post-mortem and incubated for 30 min at room temperature to retrieve sperm by float up. Afterwards, fresh sperm was analysed, and 11 samples were diluted into either a single or a double-step TEST-egg yolk-based extender to reach a final concentration of 12 % glycerol. Anatomical and histological analyses of the reproductive organs revealed that those on the right side were larger, heavier, and positioned more cranially than the left ones (P = 0.007, P = 0.003, and P = 0.0002, respectively), but sperm parameters did not differ. Successful sperm retrieval was achieved in 61.1 % of snakes, with positive correlations between seminiferous tubules length and body weight (P = 0.0004) and between seminiferous tubules length and body length (P = 0.001). Sperm-producing animals were heavier and longer compared to azoospermic ones (P = 0.04 and P = 0.04 respectively). Specifically, males longer than 130 cm and heavier than 177g have higher reproductive potential. Both cryopreservation protocols yielded comparable results, preserving sperm viability, motility, and morphology. Therefore, a single-step protocol should be preferred due to its technical simplicity. In conclusion, we described for the first time the reproductive features and characteristics of spermatozoa collected post-mortem in beauty snakes. Although the described protocols are effective, further research is warranted to optimize these techniques.}},
  articleno    = {{105184}},
  author       = {{Banchi, Penelope and Solanes Vilanova, Ferran and Cesauri, Mattia and Spanoghe, Lotte and Chiers, Koen and Van Soom, Ann and Hellebuyck, Tom and Rizzoto, Guilherme}},
  issn         = {{0011-2240}},
  journal      = {{CRYOBIOLOGY}},
  keywords     = {{Snake,Sperm,Reproductive assisted technologies,Sperm freezing,Colubridae,Reptile,Semen analysis,2-STEP DILUTION,STORAGE,SEMEN,CONSERVATION,SPERMATOZOA,PARAMETERS,SURVIVAL,EQUEX}},
  language     = {{eng}},
  pages        = {{7}},
  title        = {{Sperm cryopreservation and reproductive characteristics in the beauty snake (Elaphe taeniura) as a model for endangered Colubridae}},
  url          = {{http://doi.org/10.1016/j.cryobiol.2024.105184}},
  volume       = {{118}},
  year         = {{2025}},
}

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