Abstract
The aims of this project were to characterize tiger salamander (Ambystoma tigrinum) spermatozoa motility over time, when excreted as either milt or spermic urine prior to packaging into a spermatophore, and to determine the effect of temperature on sperm motility. A split-plot design was utilized to assess the motility of the two pre-spermatophore sample types at two temperatures, 0°C and 20°C (n = 10 for each treatment). Spermiation was induced through exogenous hormone treatment of luteinizing hormone releasing hormone analog in order to collect both milt and spermic urine, which were evaluated for motility, divided into two separate aliquots, and subsequently stored in either an ice-bath (0°C) or on the benchtop (20°C). The decay rate of sperm motility was assessed by reevaluating subsamples at 0.5, 1, 2, 3, 5, 7, and 24 hours following the initial assessment. Results showed that sperm stored at 0°C had significantly higher progressive, non-progressive, and total motility for both sperm collection types over time. An interaction was found between collection type and time, with milt exhibiting lower initial motility that was more sustainable over time, compared to spermic urine. For both milt and spermic urine, motility decreased rapidly with storage duration, indicating samples should be used as soon as possible to maximize motility for in-vitro fertilization and cryopreservation. This is the first study to describe the differences in sperm motility between milt and spermic urine from an internally fertilizing caudate and demonstrates the benefits of near freezing temperatures on sperm longevity.
Highlights
41% of amphibians are threatened due to habitat loss, overexploitation, pollution, and emerging diseases [1]
There was no effect of collection type on progressive motile” (PM) so values were pooled within temperature at each time point (Fig 3A, S1 Data)
Temperature affected Total motility (TM) with cold temperature resulting in higher motility compared to room temperature, by 8.0–19.9% for milt and 3.4–32.4% for spermic urine. This is the first study to describe the differences in sperm motility between milt and spermic urine in an internally fertilizing caudate, the tiger salamander
Summary
41% of amphibians are threatened due to habitat loss, overexploitation, pollution, and emerging diseases [1]. Only 14 reintroduction programs are considered highly successful following release of captive bred individuals [2]. Short-term storage of tiger salamander spermatozoa: The effect of collection type, temperature and time of Agriculture, Hatch project under the accession number W3173 (nifa.usda.gov). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript
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