Sperm Collection and Cryopreservation for Threatened Newt Species

ORCID

CKouba: https://orcid.org/0000-0001-8466-6077

MSU Affiliation

College of Agriculture and Life Sciences; Department of Biochemistry, Molecular Biology, Entomology, and Plant Pathology; College of Forest Resources; Department of Wildlife, Fisheries, and Aquaculture

Creation Date

2026-10-30

Abstract

The aims of this project were to transfer hormone-induced spermiation and sperm cryopreservation protocols developed in the model salamander species, Ambystoma tigrinum, to three threatened newt species. Additionally, we tested if supplementation with trehalose or thawing at different temperatures impacts post-thaw sperm parameters. Hormone stimulation protocols were applied to male Notophthalmus meridionalis (N = 10), Neurergus kaiseri (N = 5) and Tylototriton kweichowensis (N = 6) with sperm collected periodically up to 24–28 h post-spermiation dose. Samples of adequate sperm concentration (>70%) were cryopreserved in solutions of 10% Me2SO + 1% BSA with or without a 10% trehalose cryodiluent. Frozen sperm samples were thawed at either 20 °C or 40 °C and examined for post-thaw motility parameters and abnormalities in head and tail structure. The spermiation response to exogenous hormone treatment was significantly different between newt species, with a success rate of 0% for N. kaiseri, 67% for T. kweichowensis, and 100% for N. meridionalis. Sperm concentration varied with time of collection after hormone administration in both T. kweichowensis and N. meridionalis. For N. meridionalis, structural abnormalities decreased in samples collected over the 24 h period (p < 0.0001) and a thaw temperature of 40 °C resulted in higher relative total sperm motility (p < 0.0001). This is the first study to describe the cryopreservation of sperm from two newt species and demonstrates the transferability of ART developed in a salamander to two newt species.

Keywords

amphibian, cryopreservation, newt, salamander, sperm

Publication Date

5-29-2020

Publication Title

Cryobiology: International Journal of Low Temperature Biology and Medicine

Publisher

Elsevier

First Page

80

Last Page

88

Rights

© 2020 Elsevier Inc. All rights reserved.

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Digital Object Identifier (DOI)

https://doi.org/10.1016/j.cryobiol.2020.04.005