Assessment of sperm quality in palaemonid prawns using Comet assay: methodological optimization.

Area of Science:

• Marine Biology
• Ecotoxicology
• Reproductive Biology

Background:

• Sperm DNA integrity is a crucial indicator of semen quality.
• Assessing DNA integrity in crustacean spermatozoa presents unique challenges.
• The Comet assay offers a potential method for evaluating sperm quality in invertebrates.

Purpose of the Study:

• To adapt the Comet assay for evaluating DNA integrity in Palaemon serratus spermatozoa.
• To establish the Comet assay as a reliable biomarker for sperm quality in marine prawns.
• To investigate the potential of this assay for ecotoxicological assessments in Palaemonidae.

Main Methods:

• Optimized methods for sperm collection, preservation in artificial seawater (ASW), and extraction using pipetting.
• Standardized Comet assay parameters including cell lysis (1-18 hours) and DNA denaturation (15-45 minutes).
• Assessed sperm sensitivity to genotoxicants (UV-C, H2O2, MMS) and environmental contaminants (cadmium, diuron, carbamazepine).

Main Results:

• Artificial seawater (ASW) and pipetting were optimal for sperm preservation and extraction.
• 1 hour of cell lysis and 15 minutes of DNA denaturation yielded reliable Comet assay results.
• P. serratus spermatozoa showed significant DNA damage from model genotoxicants but not from tested environmental contaminants.

Conclusions:

• The adapted Comet assay is a robust and sensitive tool for assessing DNA damage in Palaemon serratus spermatozoa.
• The study highlights the potential of Palaemonidae prawns as indicators in ecogenotoxicology.
• This assay can be applied for in situ biomonitoring of sperm quality and environmental contamination impacts.