Comparison of the effect of frozen-thawed versus fresh sperm on offspring growth in tiger salamanders (Ambystoma tigrinum)

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 integration of sperm cryopreservation into amphibian captive assurance colonies provides a viable method of genetic management for threatened species. However, one fundamental concern with cryoproduced offspring is the rate of offspring development and body condition when compared to individuals produced using fresh sperm. This study analyzes the growth of salamander offspring produced using fresh or frozen-thawed sperm. Male and female A.tigrinum were administered exogenous hormones to elicit gametogenesis. Eggs were collected, fertilized with freshly collected sperm or sperm that had been cryopreserved with 10% N,N-dimethylformamide + 0.5% bovine serum albumin, and monitored through hatching. Once embryos hatched, weekly assessments of total body length were measured to track individual growth. Larvae were maintained in 100 mm x 50 mm glass dishes, with 200 mL of water per dish, and were fed a uniform diet of brine shrimp. ImageJ software was utilized to measure total length of each individual, and an analysis of variance (ANOVA) was used to determine differences in growth between larvae produced using fresh or frozen-thawed sperm. Results showed that there was no significant difference in body length between fresh larvae (n=5) and cryoproduced larvae (n=5) for week 2 (p=0.55; fresh: 1.52 ± 0.05 cm; frozen: 1.63 ± 0.05 cm) or week 3 (p=0.22, fresh: 2.01 ± 0.09 cm; frozen: 2.25 ± 0.02 cm) post-hatching. However, eggs fertilized with frozen sperm produced significantly larger larvae in week 4 than fresh sperm (p=0.02; fresh: 2.38 ± 0.08 cm; frozen: 2.74 ± 0.06 cm). Furthermore, 100% of larvae produced from fresh and frozen-thawed sperm survived through one-month post-hatching and continue to develop. This study indicates that there is minimal difference between initial growth of offspring produced using frozen-thawed or fresh sperm. The lack of deleterious effects on development supports cryopreservation as an effective tool for genetic and population management in captive colonies.

Publication Date

12-15-2022

Publication Title

Cryobiology: International Journal of Low Temperature Biology and Medicine

Publisher

Elsevier

First Page

50

Rights

© 2022 Elsevier

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Digital Object Identifier (DOI)

https://doi.org/10.1016/j.cryobiol.2022.11.160