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<p>The study of environmental DNA (eDNA) is a powerful tool for detecting and monitoring aquatic species, but sample preservation methods can critically affect analytical outcomes. Here, we examined the impact of freezing water samples—before filtration and DNA extraction—on eDNA degradation under controlled mesocosm conditions with two aquatic species: Nile tilapia (<i>Oreochromis niloticus</i>) and golden mussel (<i>Limnoperna fortunei</i>). Our findings demonstrated that freezing duration significantly reduced eDNA concentration, with a biphasic degradation pattern characterised by rapid initial loss followed by a slower degradation rate. In contrast, freezing speed did not significantly affect eDNA concentration, suggesting that the freezing process itself, rather than its speed, is the primary driver of eDNA degradation. These results highlight the importance of minimising freezing time for eDNA preservation and suggest that prioritising freezing duration over speed is sufficient in settings where rapid freezing may not be feasible. This study provides significant observations about the patterns and extent of eDNA degradation and offers practical guidelines for optimising eDNA sample processing. By understanding and mitigating the factors affecting eDNA viability, researchers can improve the accuracy and reliability of eDNA-based monitoring programmes, thereby enhancing efforts to protect and manage aquatic biodiversity.</p>

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Effects of freezing duration and speed on environmental DNA (eDNA) degradation in water samples: a controlled mesocosm study

  • 期刊论文

  • 发表日期：

    2025年7月29日

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  • Paula Valeska Stica,
  • Marcio Roberto Pie,
  • Aline Horodesky,
  • Giorgi DalPont,
  • Nathieli Cozer,
  • Vilmar Biernaski,
  • Otto Samuel Mäder Netto,
  • Andréia Szortyka,
  • Adriano Baldissera,
  • Antonio Ostrensky Neto

摘要

The study of environmental DNA (eDNA) is a powerful tool for detecting and monitoring aquatic species, but sample preservation methods can critically affect analytical outcomes. Here, we examined the impact of freezing water samples—before filtration and DNA extraction—on eDNA degradation under controlled mesocosm conditions with two aquatic species: Nile tilapia (Oreochromis niloticus) and golden mussel (Limnoperna fortunei). Our findings demonstrated that freezing duration significantly reduced eDNA concentration, with a biphasic degradation pattern characterised by rapid initial loss followed by a slower degradation rate. In contrast, freezing speed did not significantly affect eDNA concentration, suggesting that the freezing process itself, rather than its speed, is the primary driver of eDNA degradation. These results highlight the importance of minimising freezing time for eDNA preservation and suggest that prioritising freezing duration over speed is sufficient in settings where rapid freezing may not be feasible. This study provides significant observations about the patterns and extent of eDNA degradation and offers practical guidelines for optimising eDNA sample processing. By understanding and mitigating the factors affecting eDNA viability, researchers can improve the accuracy and reliability of eDNA-based monitoring programmes, thereby enhancing efforts to protect and manage aquatic biodiversity.