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Phytoplankton is a key biological group used to assess the ecological status of lakes. The classical monitoring approach relies on microscopic identification and counting of phytoplankton species, which is time-consuming and requires high taxonomic expertise. High-throughput sequencing, combined with metabarcoding, has recently demonstrated its potential as an alternative approach for plankton surveys. Several studies have confirmed the relevance of the diatom metabarcoding approach to calculate biotic indices based on species ecology. However, phytoplankton communities have not yet benefited from such validation. Here, by comparing the results obtained with the two methods (molecular and microscopic counting), we evaluated the relevance of metabarcoding approach for phytoplankton monitoring by considering different metrics: alpha diversity, taxonomic composition, community structure and a phytoplankton biotic index used to assess the trophic level of lakes. For this purpose, 55 samples were collected in four large alpine lakes (Aiguebelette, Annecy, Bourget, Geneva) during the year 2021. For each sample, a metabarcoding analysis based on two genetic markers (16S and 23S rRNA) was performed, in addition to the microscopic count. Regarding the trophic level of lakes, significant differences were found between index values obtained with the two approaches. The main hypothesis to explain these differences comes from the incompleteness, particularly at the species level, of the barcode reference library for the two genetic markers. It is therefore necessary to complete reference libraries for using such species-based biotic indices with metabarcoding data. Besides this, species richness and diversity were higher in the molecular inventories than in the microscopic ones. Moreover, despite differences in taxonomic composition of the floristic lists obtained by the two approaches, their community structures were similar. These results support the possibility of using metabarcoding for phytoplankton monitoring but in a different way. We suggest exploring alternative approaches to index development, such as a taxonomy-free approach., Competing Interests: Declaration of competing interest The achievement of this manuscript has not been influenced by competing financial, personal, or professional interests., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Assessing the vulnerability of ecosystems to biodiversity loss has become increasingly crucial in conservation and ecology research. This study proposed a methodology for measuring lake vulnerability to biodiversity loss employing an established framework that combines three components. For this, we measured the resilience (functional redundancy) and sensitivity (an index considering three characteristics of rarity) components for fish and phytoplankton communities. We also measured the exposure component of the main stressors in lakes. We then combined the three components and calculated the vulnerability index (IVCLA) using data from 255 French lakes. We found that all lakes exhibited low levels of resilience, elevated sensitivity regarding average values for fish and phytoplankton groups, and medium exposure to stressors associated with human activities. In addition, there were some discrepancies in resilience and sensitivity patterns between fish and phytoplankton groups, emphasizing the importance of considering information from multiple biological groups when assessing ecosystem vulnerability. Hydrological alterations and low water quality were key stressors related to higher lake vulnerability. Most French lakes have been classified as exhibiting moderate vulnerability. It is crucial to emphasize the potential increase in exposure risks, which could lead to even higher vulnerability levels and, subsequently, biodiversity loss in the future. The IVCLA index offers several advantages, including integrating multiple taxa groups and stressors. We recommend incorporating additional data, such as the resilience and sensitivity of the entire food web, and considering temporal responses to stressors to improve accuracy and predictive power. The IVCLA was developed with the purpose of serving as an effective tool for guiding environmental managers in designing conservation strategies and making informed decisions for lake ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)