Increasing the Yield and Cryosurvival of Spermatozoa from Rhinoceros Ejaculates Using the Enzyme Papain
The preservation of rhinoceros semen is vital for captive breeding programs. While successful collection and cryopreservation of rhinoceros semen has been reported, the volume and quality of semen produced is often low due to the high viscosity associated with ejaculates collected via electroejacula...
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MDPI AG
2022-01-01
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| Online Access: | https://www.mdpi.com/2079-7737/11/2/154 |
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| author | Jessica P. Rickard Kelsey Pool Simon P. de Graaf Timothy Portas Natalie Rourke Miriam Wiesner Thomas B. Hildebrandt Frank Göritz Robert Hermes |
| author_facet | Jessica P. Rickard Kelsey Pool Simon P. de Graaf Timothy Portas Natalie Rourke Miriam Wiesner Thomas B. Hildebrandt Frank Göritz Robert Hermes |
| author_sort | Jessica P. Rickard |
| collection | DOAJ |
| description | The preservation of rhinoceros semen is vital for captive breeding programs. While successful collection and cryopreservation of rhinoceros semen has been reported, the volume and quality of semen produced is often low due to the high viscosity associated with ejaculates collected via electroejaculation. Reducing semen viscosity would enable access to previously unusable spermatozoa from viscous fractions and could improve quality post-thaw. The enzyme papain successfully reduced the viscosity of camelid semen but has yet to be tested in wildlife species. This study assessed the influence of papain on the in vitro quality of rhinoceros spermatozoa during cryopreservation using advanced semen assessment. In experiment 1, the motility of spermatozoa from the viscous fraction of an ejaculate, either untreated or treated with papain and its inhibitor E-64 prior to cryopreservation, was assessed post-thaw. In experiment 2, spermatozoa from papain-treated viscous fractions were compared to spermatozoa frozen from untreated sperm-rich fractions pre-freeze, as well as after 0, 1.5 and 3 h of incubation post-thaw (37 °C). Papain significantly increased the quantity of spermatozoa collected from ejaculates, as well as the motility prior to freezing. Papain also improved the post-thaw motility, velocity, linearity and straightness of samples compared to sperm-rich samples, with no detriment to sperm viability, lipid membrane disorder, production of ROS or DNA integrity (<i>p</i> < 0.05). Results show the benefit of supplementing rhinoceros spermatozoa with papain prior to cryopreservation on sperm cryosurvival and demonstrates the potential of using papain to improve the success of cryopreservation protocols, not only for the rhinoceros, but also for other wildlife species. |
| first_indexed | 2024-03-09T22:36:19Z |
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| language | English |
| last_indexed | 2024-03-09T22:36:19Z |
| publishDate | 2022-01-01 |
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| series | Biology |
| spelling | doaj.art-fa85242e3f11499a8eeec40e16a915712023-11-23T18:48:53ZengMDPI AGBiology2079-77372022-01-0111215410.3390/biology11020154Increasing the Yield and Cryosurvival of Spermatozoa from Rhinoceros Ejaculates Using the Enzyme PapainJessica P. Rickard0Kelsey Pool1Simon P. de Graaf2Timothy Portas3Natalie Rourke4Miriam Wiesner5Thomas B. Hildebrandt6Frank Göritz7Robert Hermes8School of Life and Environmental Sciences, Faculty of Science, The University of Sydney, Sydney, NSW 2006, AustraliaSchool of Life and Environmental Sciences, Faculty of Science, The University of Sydney, Sydney, NSW 2006, AustraliaSchool of Life and Environmental Sciences, Faculty of Science, The University of Sydney, Sydney, NSW 2006, AustraliaZoo and Wildlife Veterinary Consultancy, Maleny, QLD 4552, AustraliaWerribee Open Range Zoo, Werribee, VIC 3030, AustraliaSalzburg Zoo Hellbrunn, 5081 Anif, AustriaDepartment of Reproductive Management, Leibniz Institute for Zoo and Wildlife Research, 10315 Berlin, GermanyDepartment of Reproductive Management, Leibniz Institute for Zoo and Wildlife Research, 10315 Berlin, GermanyDepartment of Reproductive Management, Leibniz Institute for Zoo and Wildlife Research, 10315 Berlin, GermanyThe preservation of rhinoceros semen is vital for captive breeding programs. While successful collection and cryopreservation of rhinoceros semen has been reported, the volume and quality of semen produced is often low due to the high viscosity associated with ejaculates collected via electroejaculation. Reducing semen viscosity would enable access to previously unusable spermatozoa from viscous fractions and could improve quality post-thaw. The enzyme papain successfully reduced the viscosity of camelid semen but has yet to be tested in wildlife species. This study assessed the influence of papain on the in vitro quality of rhinoceros spermatozoa during cryopreservation using advanced semen assessment. In experiment 1, the motility of spermatozoa from the viscous fraction of an ejaculate, either untreated or treated with papain and its inhibitor E-64 prior to cryopreservation, was assessed post-thaw. In experiment 2, spermatozoa from papain-treated viscous fractions were compared to spermatozoa frozen from untreated sperm-rich fractions pre-freeze, as well as after 0, 1.5 and 3 h of incubation post-thaw (37 °C). Papain significantly increased the quantity of spermatozoa collected from ejaculates, as well as the motility prior to freezing. Papain also improved the post-thaw motility, velocity, linearity and straightness of samples compared to sperm-rich samples, with no detriment to sperm viability, lipid membrane disorder, production of ROS or DNA integrity (<i>p</i> < 0.05). Results show the benefit of supplementing rhinoceros spermatozoa with papain prior to cryopreservation on sperm cryosurvival and demonstrates the potential of using papain to improve the success of cryopreservation protocols, not only for the rhinoceros, but also for other wildlife species.https://www.mdpi.com/2079-7737/11/2/154rhinoviscousfreezingconservationflow cytometryCASA |
| spellingShingle | Jessica P. Rickard Kelsey Pool Simon P. de Graaf Timothy Portas Natalie Rourke Miriam Wiesner Thomas B. Hildebrandt Frank Göritz Robert Hermes Increasing the Yield and Cryosurvival of Spermatozoa from Rhinoceros Ejaculates Using the Enzyme Papain Biology rhino viscous freezing conservation flow cytometry CASA |
| title | Increasing the Yield and Cryosurvival of Spermatozoa from Rhinoceros Ejaculates Using the Enzyme Papain |
| title_full | Increasing the Yield and Cryosurvival of Spermatozoa from Rhinoceros Ejaculates Using the Enzyme Papain |
| title_fullStr | Increasing the Yield and Cryosurvival of Spermatozoa from Rhinoceros Ejaculates Using the Enzyme Papain |
| title_full_unstemmed | Increasing the Yield and Cryosurvival of Spermatozoa from Rhinoceros Ejaculates Using the Enzyme Papain |
| title_short | Increasing the Yield and Cryosurvival of Spermatozoa from Rhinoceros Ejaculates Using the Enzyme Papain |
| title_sort | increasing the yield and cryosurvival of spermatozoa from rhinoceros ejaculates using the enzyme papain |
| topic | rhino viscous freezing conservation flow cytometry CASA |
| url | https://www.mdpi.com/2079-7737/11/2/154 |
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