Evaluation of dimethylformamide (DMF) and Trehalose as cryoprotectants in African penguin Spheniscus demersus semen cryopreservation

evaluation-of-dimethylformamide-(dmf)-and-trehalose-as-cryoprotectants-in-african-penguin-spheniscus-demersus-semen-cryopreservation
Evaluation of dimethylformamide (DMF) and Trehalose as cryoprotectants in African penguin Spheniscus demersus semen cryopreservation

References

  1. De Wit, P. et al. EAZA Penguin Taxon Advisory Group Regional Collection Plan – First Edition (EAZA Executive Office, 2021).

  2. Hacker, F. et al. Effect of environmental variables on African penguin vocal activity: Implications for acoustic censusing. Biology 12,1191(2023).

  3. BirdLife, I. Spheniscus demersus. The IUCN Red List of Threatened Species 2024: e.T22697810A256021744. (2024). (2024). https://dx.doi.org/10.2305/IUCN.UK.2024-2.RLTS.T22697810A256021744.en (2025).

  4. Pichegru, L. et al. A decade of implementing the biodiversity management plan for African penguins – successes, failures and lessons learnt. J. Nat. Conserv. 86, 126919 (2025).

    Google Scholar 

  5. Ancel, A., Beaulieu, M. & Gilbert, C. The different breeding strategies of penguins: A review. C R Biol. 336, 1–12 (2013).

    Google Scholar 

  6. Hockey, P. The African Penguin – A natural historyStruik, (2001).

  7. Dreyer, S. et al. Factors in the decline of the African penguin: are contaminants of emerging concern (CECs) a potential new age stressor? Mar. Pollut Bull. 206, 116688 (2024).

    Google Scholar 

  8. Crawford, R. J. M. et al. Food limitation of seabirds in the Benguela ecosystem and management of their prey base. NJE 6A, 1–13 (2022).

    Google Scholar 

  9. Pichegru, L. et al. Maritime traffic trends around the Southern tip of Africa – Did marine noise pollution contribute to the local penguins’ collapse? Sci. Total Environ. 849, 157878 (2022).

    Google Scholar 

  10. Daniels, R. Save the African Penguin. SANCCOB. https://sanccob.co.za/news/save-the-african-penguin/ (2022).

  11. Bos, C., Schad, K. & Fienieg, E. Long-term Management Plan for the African Penguin (Spheniscus demersus) European Endangered Species Programme (EEP) (Artis Royal Zoo, 2016).

  12. Barham, P. J., Underhill, L. G., Crawford, R. J. M. & Leshoro, T. M. Differences in breeding success between African Penguins (Spheniscus demersus) that were and were not oiled in the MV treasure oil-spill in 2000. Emu 107, 7–13 (2007).

    Google Scholar 

  13. Brühl, J. & Turpie, J. An Assessment of the Value of African Penguins (Report prepared by Anchor Environmental Consultants for the Endangered Wildlife Trust and the Department of Forestry, Fisheries and the Environment, 2024).

  14. Sherley, R. B. et al. The African Penguin Spheniscus demersus should be considered critically endangered. Ostrich 95 (3), 181–187 (2024).

    Google Scholar 

  15. McInnes, A. M. et al. Commercial fishery no-take zones for African Penguins minimize fisheries losses at the expense of conservation gains. ICES J. Mar. Sci. 81 (8), 1632–1646 (2024).

    Google Scholar 

  16. Blesbois, E. & Brillard, J. P. Specific features of in vivo and in vitro sperm storage in birds. Animal 1, 1472–1481 (2007).

    Google Scholar 

  17. Rakha, B. A. et al. Use of dimethylsulfoxide for semen cryopreservation in Indian red jungle fowl (Gallus Gallus murghi). Theriogenology 122, 61–67 (2018).

    Google Scholar 

  18. Thananurak, P., Chuaychu-noo, N., Phasuk, Y. & Vongpralub, T. Comparison of TNC and standard extender on post-thaw quality and in vivo fertility of Thai native chicken sperm. Cryobiology 92, 197–202 (2020).

    Google Scholar 

  19. Onur Özkök, A., Esin, B. & Akal, E. Sperm cryopreservation in Canaries to protect endangered Songbird species: comparison of different cryoprotectants. Vet. Med. Sci. 10(6), e70101 (2024).

    Google Scholar 

  20. Kowalczyk, A. & Łukaszewicz, E. Simple and effective methods of freezing capercaillie (Tetrao urogallus L.) semen. PLoS ONE 10, e0116797 (2015).

    Google Scholar 

  21. O’Brien, J. K., Oehler, D. A., Malowski, S. P. & Roth, T. L. Semen Collection, Characterization, and cryopreservation in a Magellanic Penguin (Spheniscus magellanicus). Zoo Biol. 18, 199–214 (1999).

    Google Scholar 

  22. Santiago-Moreno, J. et al. Semen cryopreservation in black-footed (Spheniscus demersus) and Gentoo (Pygoscelis papua) penguins: effects of thawing temperature on semen characteristics. Anim. Reprod. Sci. 200, 60–66 (2019).

    Google Scholar 

  23. O’Brien, J. K., Steinman, K. J., Montano, G. A., Dubach, J. M. & Robeck, T. R. Chicks produced in the Magellanic Penguin (Spheniscus magellanicus) after cloacal insemination of frozen-thawed semen. Zoo Biol. 35, 326–338 (2016).

    Google Scholar 

  24. Marti-Colombas, M. et al. Optimization of semen cryopreservation in black-footed (Spheniscus demersus) and Gentoo (Pygoscelis papua) Penguins using dimethylacetamide and dimethylsulfoxide. Anim. Reprod. Sci. 237, 106933 (2022).

    Google Scholar 

  25. O’Brien, J. K. & Robeck, T. R. Semen characterization, seasonality of production, and in vitro sperm quality after chilled storage and cryopreservation in the King Penguin (Aptenodytes patagonicus). Zoo Biol. 33, 99–109 (2014).

    Google Scholar 

  26. Mafunda, P. S. Aspects of the Reproduction of Male and Female African Penguins (Spheniscus demersus) with Special Reference To Sperm Biology and Cryopreservation (University of the Western Cape, 2018).

  27. Zong, Y. et al. Chicken sperm cryopreservation: review of techniques, freezing damage, and freezability mechanisms. Agriculture 13 (2), 445 (2023).

    Google Scholar 

  28. Blanco, J. M., Long, J. A., Gee, G., Wildt, D. E. & Donoghue, A. M. Comparative cryopreservation of avian spermatozoa: benefits of non-permeating osmoprotectants and ATP on Turkey and crane sperm cryosurvival. Anim. Reprod. Sci. 123, 242–248 (2011).

    Google Scholar 

  29. Janosikova, M. et al. New approaches for long-term conservation of rooster spermatozoa. Poult. Sci. 102 (2), 102386 (2023).

    Google Scholar 

  30. Rakha, B. A. et al. Effect of dimethylformamide on sperm quality and fertilizing ability of Indian red jungle fowl (Gallus Gallus murghi). Theriogenology 149, 55–61 (2020).

    Google Scholar 

  31. Polsang, S. et al. Effects of extenders and cryoprotectants on cryopreservation of Thai red junglefowl (Gallus Gallus Gallus) spermatozoa. Cryobiology 106, 48–54 (2022).

    Google Scholar 

  32. Thananurak, P., Vongpralup, T., Sittikasamkit, C. & Sakwiwatkul, K. Optimization of Trehalose concentration in semen freezing extender in Thai native chicken semen. Thai J. Vet. Med. 46 (2), 287–294 (2016).

    Google Scholar 

  33. Thananurak, P., Chuaychu-Noo, N. & Vongpralub, T. Freezability and fertility of Thai native chicken semen in different diluents. Thai J. Vet. Med. 47 (4), 551–556 (2017).

    Google Scholar 

  34. Partyka, A., Łukaszewicz, E. & Niżański, W. Flow cytometric assessment of fresh and frozen-thawed Canada Goose (Branta canadensis) semen. Theriogenology 76, 843–850 (2011).

    Google Scholar 

  35. Flores Huarco, N. H. et al. Use of dimethylformamide to cryopreserve alpaca semen previously incubated with collagenase. Reprod. Dom Anim. 56 (11), 1387–1397 (2021).

    Google Scholar 

  36. Bruno, S. L. et al. Comparison of different cryoprotectants for freezing Donkey (Equus asinus) semen. J. Equine Vet. Sci. 136, 105069 (2024).

    Google Scholar 

  37. Stanishevskaya, O., Silyukova, Y., Pleshanov, N. & Kurochkin, A. Role of mono-and disaccharide combination in cryoprotective medium for rooster semen to ensure cryoresistance of spermatozoa. Molecules 26(19), 5920 (2021).

    Google Scholar 

  38. Ahmad, E. & Aksoy, M. Trehalose as a cryoprotective agent for the sperm cells: A mini review. AHPH 1(2),123–129 (2012).

    Google Scholar 

  39. Zhu, Z., Fan, X., Pan, Y., Lu, Y. & Zeng, W. Trehalose improves rabbit sperm quality during cryopreservation. Cryobiology 75, 45–51 (2017).

    Google Scholar 

  40. Gholami, D. et al. Beneficial effects of Trehalose and gentiobiose on human sperm cryopreservation. PLoS ONE 18(4), e0271210 (2023).

    Google Scholar 

  41. Jia, B. et al. Trehalose modifies the protein profile of Ram spermatozoa during cryopreservation. Theriogenology 171, 21–29 (2021).

    Google Scholar 

  42. Murray, A., Kilbride, P. & Gibson, M. I. Trehalose in cryopreservation. Applications, mechanisms and intracellular delivery opportunities. RSC Med. Chem. 15, 2980–2995 (2024).

    Google Scholar 

  43. Borecki, P., Mucha, A., Niżański, W. & Partyka, A. Factors determining semen sample collection and semen quality parameters in African Penguins Spheniscus demersus. Sci. Rep. 14, 24261. https://doi.org/10.1038/s41598-024-76303-2 (2024).

    Google Scholar 

  44. Mosca, F. et al. Combined effect of permeant and non-permeant cryoprotectants on the quality of frozen/thawed chicken sperm. Cryobiology 73, 343–347 (2016).

    Google Scholar 

  45. Humann-Guilleminot, S. et al. Sperm collection in Black-legged Kittiwakes and characterization of sperm velocity and morphology. Avian Res. 9, 24 (2018).

    Google Scholar 

  46. Matsuzaki, M. et al. Longer and faster sperm exhibit better fertilization success in Japanese quail. Poult. Sci. 100 (4), 100980 (2021).

    Google Scholar 

  47. Mocé, E., Grasseau, I. & Blesbois, E. Cryoprotectant and freezing-process alter the ability of chicken sperm to acrosome React. Anim. Reprod. Sci. 122 (3–4), 359–366. https://doi.org/10.1016/j.anireprosci.2010.10.010 (2010).

    Google Scholar 

  48. Olexikova, L., Miranda, M., Kulikova, B., Baláži, A. & Chrenek, P. Cryodamage of plasma membrane and acrosome region in chicken sperm. Anat. Histol. Embryol. 48 (1), 33–39 (2019).

    Google Scholar 

  49. Fu, J. et al. Effect of DMSO combined with Trehalose on cryopreservation of Goose semen. J. Appl. Anim. Res. 51 (1), 84–91 (2023).

    Google Scholar 

  50. Hai, E., Li, B., Zhang, J. & Zhang, J. Sperm freezing damage: the role of regulated cell death. Cell. Death Discov. 10, 239. https://doi.org/10.1038/s41420-024-02013-3 (2024).

    Google Scholar 

  51. Said, T. M., Gaglani, A. & Agarwal, A. Implication of apoptosis in sperm cryoinjury. Reprod. BioMed. Online. 21, 456–462. https://doi.org/10.1016/j.rbmo.2010.05.011 (2010).

    Google Scholar 

  52. Lee, Y. A. et al. Cryopreservation in Trehalose preserves functional capacity of murine spermatogonial stem cells. PLoS ONE. 8 (1). https://doi.org/10.1371/journal.pone.0054889 (2013). e54889.

  53. Zini, A., San Gabriel, M. & Baazeem, A. Antioxidants and sperm DNA damage: A clinical perspective. J Assist. Reprod Genet. 26 (8), 427–432. https://doi.org/10.1007/s10815-009-9343-5 (2009).

    Google Scholar 

  54. Fleming, S. D. & Thomson, L. K. The oxidative stress of human sperm cryopreservation. Antioxidants 14 (4), 402. https://doi.org/10.3390/antiox14040402 (2025).

    Google Scholar 

  55. Madeddu, M. et al. Differences in semen freezability and intracellular ATP content between the rooster (Gallus Gallus domesticus) and the barbary Partridge (Alectoris barbara). Theriogenology 74 (6), 1010–1018 (2010).

    Google Scholar 

  56. Gliozzi, T. M., Zaniboni, L. & Cerolini, S. DNA fragmentation in chicken spermatozoa during cryopreservation. Theriogenology 75 (9), 1613–1622 (2011).

    Google Scholar 

  57. Burrows, W. H. & Quinn, J. P. A method of obtaining spermatozoa from the domestic fowl. Poult. Sci. 14, 253–254 (1935).

    Google Scholar 

  58. Blesbois, E. et al. Predictors of success of semen cryopreservation in chickens. Theriogenology 69 (2), 252–261 (2008).

    Google Scholar 

  59. Bernal, B. et al. Catalase, superoxide dismutase and butylated hydroxytoluene benefit mid-term storage of red-legged Partridge sperm (Alectoris rufa). Br. Poult. Sci. 66 (1), 131–138. https://doi.org/10.1080/00071668.2024.2394185 (2024).

    Google Scholar 

  60. R Core Team. R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL (2024). https://www.R-project.org/

  61. Tomczak, M. & Tomczak, E. The need to report effect size estimates revisited. An overview of some recommended measures of effect size. TSS 1 (21), 19–25 (2014).

    Google Scholar 

  62. Revelle, W. psych: Procedures for Personality and Psychological Research, Version = 2.0.12Northwestern University, (2020). Available at: https://CRAN.R-project.org/package=psych

  63. Chessel, D., Dufour, A. & Thioulouse, J. The ade4 Package – I: One-Table methods. R News. 4 (1), 5–10 (2004). https://cran.r-project.org/doc/Rnews/

    Google Scholar 

  64. Kassambara, A. & Mundt, F. factoextra: Extract and Visualize the Results of Multivariate Data Analyses, Version 1.0.7 (2020). Available at: https://CRAN.R-project.org/package=factoextra

Download references