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Introduction
Results
The percentage of not assigned reads was lower at the higher size-fractions. While maxillopoda
predominated at those size fractions, a more diverse assemblage characterized the 0.22-20 m one.
Copepods represented 48.6, 36 and 2.3% while phytoplankton groups <0.1, 1.6 and 32.7% for each
size-fraction respectively.
The CCA explained 57.7% of variance. Main environmental factors were salinity and date. While a
reduced number of brackish water species, such as the copepods A. tonsa and Calanipeda aquaedulcis
characterized the 30 community, a higher number of OTUs encompassing mostly neritic taxa
conformed the 35 water mass.
Correlations were signicant in most of the cases and with no noticeable eect when comparing
against microscopy-based counts or biomass.
Whilst similar relative abundances were found for A. tonsa in the 30 water mass by both approaches
(Fig 4a), it was only detected by metabarcoding in the 35 salinity (Fig 4b). Regarding P. marinus,
detection was favorable to metabarcoding in six out of eight cases meanwhile only in two its presence
was detected by microscopy (Fig 4c, d).
Figure 1. Proportion of taxonomic groups in each sample based on the metabarcoding approach.
Environmental Sample
(e.g. Filtered Seawater)
DNA
extraction
Taxonomic Compositon
Bioinformatics
PCR
Amplication
Sequencing
Figure 4. Comparison of metabarcoding and microscopy when assessing abundances for NIS.
Blue arrows and green ellipses indicate temporal and spatial cycles respectively.
Conclusions
The somewhat reduced performance of this approach for the lowest size fractions is mainly related to
18s V9 database incompleteness for these organisms. This highlights that DNA-barcoding is necessary
and complementary to metabarcoding [6].
Metabarcoding replicated the Bilbao estuary plankton community temporal and spatial patterns.
The lack of correlation between relative abundances could be explained by technical biases introduced
during the DNA extraction [7] or PCR amplication step [8]. The Copy Number Variation (CNV)
associated to multi-copy genes, such as rRNA ones, has been suggested as one of the main factors
limiting the quantitative value of metabarcoding [9]. In the meantime, metabarcoding targeting multicopy genes will remain as a semi-quantitative approach [10].
The present study demonstrated the suitability of metabarcoding for early detection of NIS at
extremely low abundances (Fig 4), conrming previous studies [11, 12]. The reasons behind this are a)
the ability to analyze higher sample volumes and b) the capacity to take into account individuals at
any life stage, such as eggs or nauplius larvae.
All of this suggests that metabarcoding could be a powerful tool, if implemented in plankton
monitoring, for early detection of NIS or plankton biodiversity shifts.
Bibliography
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