Unlocking river biofilm microbial diversity: a comparative analysis of sequencing technologies

Anderson, Meri A.J.; Thorpe, Amy C.; Busi, Susheel Bhanu; Gweon, Hyun Soon; Warren, Jonathan; Walsh, Kerry; Read, Daniel S.

Unlocking river biofilm microbial diversity: a comparative analysis of sequencing technologies

Anderson, Meri A.J. ORCID: https://orcid.org/0009-0007-8889-9485; Thorpe, Amy C. ORCID: https://orcid.org/0000-0003-0210-2767; Busi, Susheel Bhanu ORCID: https://orcid.org/0000-0001-7559-3400; Gweon, Hyun Soon ORCID: https://orcid.org/0000-0002-6218-6301; Warren, Jonathan ORCID: https://orcid.org/0000-0003-3381-3852; Walsh, Kerry ORCID: https://orcid.org/0000-0001-8619-8895; Read, Daniel S. ORCID: https://orcid.org/0000-0001-8546-5154. 2025 Unlocking river biofilm microbial diversity: a comparative analysis of sequencing technologies. Molecular Ecology Resources, e70075. 12, pp. 10.1111/1755-0998.70075

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Official URL: https://doi.org/10.1111/1755-0998.70075

Abstract/Summary

Freshwater ecosystems are under increasing pressure from pollution, habitat degradation and climate change, highlighting the need for reliable biomonitoring approaches to assess ecosystem health and identify the causes of biodiversity and ecosystem service loss. Characterisation of freshwater microbiomes has the potential to be an important tool for understanding freshwater ecology, ecosystem health and ecosystem function. High‐throughput sequencing technologies, such as Illumina short‐read and Pacific Biosciences long‐read sequencing, are widely used for microbial community analysis. However, the relative performance of these approaches for monitoring freshwater microbiomes has not been well explored. In this study, we compared the performance of long‐ and short‐read sequencing approaches to assess archaeal and bacterial diversity in 42 river biofilm samples across seven distinct river sites in England by targeting the 16S ribosomal RNA gene. Our findings demonstrated that longer reads generated by PacBio sequencing provide a higher taxonomic resolution, enabling the classification of taxa that remained unassigned in the short‐read Illumina datasets. This enhanced resolution is particularly beneficial for biodiversity assessments because it improves species‐level identification, which is crucial for ecological monitoring. Despite this, both sequencing methods produced comparable bacterial community structures regarding taxon relative abundance, suggesting that the sequencing approach does not profoundly affect the comparative assessment of community composition. However, while Illumina offers higher throughput and cost efficiency, PacBio's ability to resolve complex microbial communities highlights its potential for studies requiring precise taxonomic identification.

Item Type:

Publication - Article

Digital Object Identifier (DOI):

10.1111/1755-0998.70075

UKCEH and CEH Sections/Science Areas:

Environmental Pressures and Responses (2025-)

ISSN:

1755-098X

Additional Information:

Open Access paper - full text available via Official URL link.

Additional Keywords:

bacterial community composition, epilithic biofilms, long-read sequencing, short-read sequencing