Integrating visual and molecular approaches for fish eggs analysis: A study on formaldehyde fixation and storage procedures

Accurate taxonomic classification and developmental stage determination of fish eggs are crucial for ecological monitoring, conservation efforts, and stock assessments. Traditional methods for fish and fisheries rely on visual examination of morphological traits, but they face challenges due to species overlap especially for early stages. Molecular tools, such as DNA barcoding, offer higher resolution in taxonomic identification but may not provide developmental stage information. This study explores the effectiveness of different formaldehyde fixation concentrations and storage procedures on fish eggs collected from Lofoten, Norway, for both visual and molecular analysis. Visual analysis successfully identified developmental stage for all fixation solutions. Molecular barcoding using the 16S rRNA gene identified up to 100% of eggs at the species level, with decreasing success rates over time when stored in formaldehyde fixation. The highest DNA barcoding success rates were accomplished using 4% formaldehyde fixation for 12- or 24-h following transfer to ethanol. Using 0.5% and 1% formaldehyde fixation up to 8 weeks also resulted in high DNA success rates, but results deteriorated with increasing storage time. This study provides valuable insights for integrating visual and molecular methods for fish egg analysis, with practical implications for sample preservation during marine surveys.