S.D. Mims | USGS Science

USGS2008

Viability and fertilizing capacity of cryopreserved sperm from three North American acipenseriform species: A retrospective study

Populations of sturgeon across the globe are threatened due to unregulated harvest and habitat loss, and the status varies among species across North America. Ready access to viable and functional sperm would contribute to recovery programmes for these species. In this study, we examined the motility, viability (cell membrane integrity) of cryopreserved sperm from three North American acipenseriform species and fertilizing capacity. Milt samples were collected from captive shortnose sturgeon (Acipenser brevirostrum), wild paddlefish (Polyodon spathula) and pallid sturgeon (Scaphirhynchus albus) and cryopreserved using combinations of Modified Tsvetkova's (MT) extender, Original Tsvetkova's extender, and modified Hanks' balanced salt solution, along with the cryoprotectants methanol (MeOH) or dimethyl sulfoxide (DMSO). A dual-staining technique using the fluorescent stains SYBR-14 and propidium iodide was employed with flow cytometry to determine the percentages of spermatozoa that were viable by virtue of having intact membranes. The percentage of viable spermatozoa ranged from 5% to 12% in shortnose sturgeon, 30-59% in paddlefish, and 44-58% in pallid sturgeon. In the first experiment with shortnose sturgeon sperm, methanol allowed for higher values for dependent variables than did DMSO, and sperm viability generally correlated with post-thaw motility. However, fertilization rate, neurulation, or hatching rates were independent from these factors. In the second experiment with shortnose sturgeon, 5% MeOH combined with MT yielded higher values for all parameters tested than the other combinations: viability was correlated with motility, fertilization rate, and hatching rate. Overall, viability and post-thaw motility was not affected by the use of hyperosmotic extenders (OT) or cryoprotectants (DMSO), but their use decreased fertilization percentages. For paddlefish sperm (experiment 3), MT combined with 10% MeOH was clearly a good choice for cryopreservation; viability and motility results were correlated, but independent of fertilization. For pallid sturgeon sperm (experiment 4), MT with 5-10% MeOH showed significantly higher sperm quality and fertilization parameters. Membrane integrity can be used as a predictor of fertilization by cryopreserved sperm, however additional sperm quality parameters, supplementary to motility and membrane integrity, would be useful in the refining and optimizing cryopreservation protocols with acipenseriform sperm. ?? 2008 Blackwell Verlag, Berlin.

Journal of Applied Ichthyology↗

USGS2000

Biomarker response and health of polychlorinated biphenyl- and chlordane-contaminated paddlefish from the Ohio River Basin, USA

Fifty paddlefish ( Polyodon spathula ) collected from two sites on the Ohio River and from one site on the Cumberland River, USA, were examined to determine gonad polychlorinated biphenyl (PCB) and chlordane concentrations, amounts of plasma sex steroids (testosterone and estradiol), hepatic microsomal ethoxyresorufin- O -deethylase (EROD) activity, and the presence of immunoreactive cytochrome P450 1A (CYP1A) protein. Percent hatch and liver, spleen, and kidney histology were also determined. Gonad PCB and chlordane concentrations were significantly higher in Ohio River paddlefish than in Cumberland River paddlefish. Gonad PCB and chlordane concentration and gonad percent lipid were correlated in Ohio River paddlefish. Five of 10 Ohio River egg samples exceeded the Food and Drug Administration's action limit for chlordane (0.30 μg/g). Polychlorinated biphenyl congener-specific analysis detected predominantly the tetra-, penta-, and hexachlorobiphenyls in paddlefish testes. Plasma testosterone levels were significantly lower in males collected from the upper Ohio River site than those collected from the lower part of the river. There was no measurable hepatic microsomal EROD activity in any of the 50 paddlefish collected from the three sites. Western blotting analysis confirmed that a rabbit antitrout CYP1A1 IgG antibody did not recognize a CYP1A protein in paddlefish liver microsomes. Percent hatch was not significantly different in eggs collected from the Cumberland (88–96%) and Ohio Rivers (90–95%). Histological analysis of liver, spleen, and kidney detected the presence of hepatic steatosis and hemosiderosis, splenic lymphoid cell depletion, and hyperplasia of interrenal and chromaffin tissues. Immunosuppression, hepatic metabolic disorders, and altered neuroendocrine function may be occurring in Ohio River paddlefish. Results presented here suggest that organochlorine exposure may be jeopardizing the long-term health of Ohio River paddlefish and that additional investigation of contaminant effects on immune system function and hormone levels in paddlefish is warranted.

Alabama, Georgia, Illinois, Indiana, Kentucky, Mar↗