Are you filtering enough? Unveiling the impact of water sample volumes on eukaryotic microbial community in freshwater lakes

The appropriate volume of filtered water samples is crucial for accurately reflecting microbial community characteristics. Previous research has shown that changes in water sample volumes significantly affect bacterial diversity and the relative abundance of dominant taxa in marine environments. However, the specific impact on freshwater microbial diversity and community structure, particularly for eukaryotic microorganisms, remains unclear. To address this gap, we collected water samples from eutrophic Lake Taihu and mesotrophic Lake Bosten. We filtered the samples through 0.2 μm filters using volumes ranging from 0.1 to 2 L for Lake Taihu and 0.1 to 3.2 L for Lake Bosten. Analysis of 18S rRNA gene amplicons revealed that variations in filtered water sample volume significantly affected the diversity, community structure, and composition of eukaryotic microorganisms. In light of these findings, we recommend using a filtered water sample volume of approximately 0.2 L for eutrophic aquatic ecosystems and 0.8 L for mesotrophic aquatic ecosystems in microbial analyses. These recommendations underscore the importance of selecting the appropriate filtered water sample volume to ensure representative and reliable results in the study of eukaryotic microorganisms in freshwater ecosystems.