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DNA metabarcoding assessment of Neotropical ichthyoplankton communities is marker-dependent.

ABSTRACT: The study of ichthyoplankton is paramount to understanding fish assemblages' reproductive dynamics. DNA metabarcoding has been applied as a rapid, cost-effective, and accurate taxonomy tool, allowing the identification of multiple individuals simultaneously. However, there remain significant challenges when using DNA metabarcoding, such as molecular marker choice according to the taxonomic resolution and length of the fragment to be sequenced, primer bias, incomplete reference databases, and qualitative inference incongruences. Here, 30 ichthyoplankton pools collected from a Neotropical river were identified at a molecular level using DNA metabarcoding to compare the resolution, sensibility, specificity, and relative read abundance (RRA) recovery of three molecular markers: the standard COI fragment (650 pb, with each end analyzed individually) and two short 12S rRNA genes markers (≅200 bp - NeoFish and MiFish markers). The combined use of the three markers increased the genera detection rates by 25%-87.5%, allowing an increased taxonomic coverage and robust taxonomic identification of complex Neotropical ichthyoplankton communities. RRA is marker-dependent, indicating caution is still needed while inferring species abundance based on DNA metabarcoding data when using PCR-dependent protocols.

SUBMITTER: Teixeira DF

PROVIDER: S-EPMC10587807 | biostudies-literature | 2023 Oct

REPOSITORIES: biostudies-literature

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DNA metabarcoding assessment of Neotropical ichthyoplankton communities is marker-dependent.

Teixeira Daniel Fonseca DF Hilário Heron Oliveira HO Santos Gilmar Bastos GB Carvalho Daniel Cardoso DC

Ecology and evolution 20231020 10

The study of ichthyoplankton is paramount to understanding fish assemblages' reproductive dynamics. DNA metabarcoding has been applied as a rapid, cost-effective, and accurate taxonomy tool, allowing the identification of multiple individuals simultaneously. However, there remain significant challenges when using DNA metabarcoding, such as molecular marker choice according to the taxonomic resolution and length of the fragment to be sequenced, primer bias, incomplete reference databases, and qua ...[more]

PMID: 37869433

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Project description:Drifting fish eggs and larvae (ichthyoplankton) can be identified to species using DNA metabarcoding, thus allowing for post hoc community analyses at a high taxonomic resolution. We undertook a regional-scale study of ichthyoplankton distribution along the east coast of South Africa, focused on the contrasting environments of the tropical Delagoa and subtropical Natal Ecoregions, and on exposed and sheltered shelf areas. Zooplankton samples were collected with tow nets at discrete stations along cross-shelf transects (20-200 m depth) spaced along a latitudinal gradient that incorporates a known biogeographical boundary. Metabarcoding detected 67 fish species, of which 64 matched prior distribution records of fishes from South Africa, with the remaining three known from the Western Indian Ocean. Coastal, neritic and oceanic species were present, from epi- and mesopelagic to benthopelagic and benthic adult habitats. By family, Myctophidae (10 species), Carangidae, Clupeidae, Labridae (each with 4 species) and Haemulidae (3 species) were most speciose. Ichthyoplankton community composition varied significantly with latitude, distance to coast, and distance to the shelf edge. Small pelagic fishes had the highest frequency of occurrence: Engraulis capensis, Emmelichthys nitidus and Benthosema pterotum increased in frequency towards the north, whereas Etrumeus whiteheadi increased towards the south. Chub mackerel Scomber japonicus accounted for most variability related to distance from the coast, whilst African scad Trachurus delagoa correlated with distance to the shelf edge. Dissimilarity between communities in the Delagoa and Natal Ecoregions was 98-100%, whereas neighbouring transects located within the sheltered KwaZulu-Natal Bight had lower dissimilarity (56-86%). Onshore transport of ichthyoplankton by Agulhas Current intrusions plausibly explained the abundance of mesopelagic species over the shelf. Metabarcoding followed by community analysis revealed a latitudinal gradient in the ichthyoplankton, associations with coastal and shelf-edge processes, and evidence of a spawning area in the sheltered KwaZulu-Natal Bight.

Project description:For DNA metabarcoding to attain its potential as a community assessment tool, we need to better understand its performance versus traditional morphological identification and work to address any remaining performance gaps in incorporating DNA metabarcoding into community assessments. Using fragments of the 18S nuclear and 16S mitochondrial rRNA genes and two fragments of the mitochondrial COI marker, we examined the use of DNA metabarcoding and traditional morphological identification for understanding the diversity and composition of crustacean zooplankton at 42 sites across western Lake Superior. We identified 51 zooplankton taxa (genus or species, depending on the finest resolution of the taxon across all identification methods), of which 17 were identified using only morphological traits, 13 using only DNA and 21 using both methods. The taxa found using only DNA metabarcoding included four species and one genus-level identification not previously known to occur in Lake Superior, the presence of which still needs to be confirmed. A substantial portion of taxa that were identified to genus or species by morphological identification, but not identified using DNA metabarcoding, had zero ("no record") or ≤ 2 ("underrepresented records") reference barcodes in the BOLD or NCBI databases (63% for COI, 80% for 16S, 74% for 18S). The two COI marker fragments identified the most genus- and species-level taxa, whereas 18S was the only marker whose family-level percent sequence abundance patterns showed high correlation to composition patterns from morphological identification, based on a NMDS analysis of Bray-Curtis similarities. Multiple replicates were collected at a subset of sites and an occupancy analysis was performed, which indicated that rare taxa were more likely to be detected using DNA metabarcoding than traditional morphology. Our results support that DNA metabarcoding can augment morphological identification for estimating zooplankton diversity and composition of zooplankton over space and time, but may require use of multiple markers. Further addition of taxa to reference DNA databases will improve our ability to use DNA metabarcoding to identify zooplankton and other invertebrates in aquatic surveys.

Project description:BackgroundFreshwater ecosystems, such as streams, are facing increasing pressures from agricultural land use and recent literature stresses the importance of robust biomonitoring to detect trends in insect decline globally. Aquatic insects and other macroinvertebrates are often used as indicators of ecological condition in freshwater biomonitoring programs; however, these diverse groups can present challenges to morphological identification and coarse-level taxonomic resolution can mask patterns in community composition. Here, we incorporate molecular identification (DNA metabarcoding) into a stream biomonitoring sampling design to explore the diversity and variability of aquatic macroinvertebrate communities at small spatial scales. While individual stream reaches can be very heterogenous, most community ecology studies focus on larger, landscape-level patterns of community composition. A high degree of community variability at the local scale has important implications for both biomonitoring and ecological research, and the incorporation of DNA metabarcoding into local biodiversity assessments will inform future sampling protocols.ResultsWe sampled twenty streams in southern Ontario, Canada, for aquatic macroinvertebrates across multiple time points and assessed local community variability by comparing field replicates taken ten meters apart within the same stream. Using bulk-tissue DNA metabarcoding, we revealed that aquatic macroinvertebrate communities are highly diverse at small spatial scales with unprecedented levels of local taxonomic turnover. We detected over 1600 Operational Taxonomic Units (OTUs) from 149 families, and a single insect family, the Chironomidae, contained over one third of the total number of OTUs detected in our study. Benthic communities were largely comprised of rare taxa detected only once per stream despite multiple biological replicates (24-94% rare taxa per site). In addition to numerous rare taxa, our species pool estimates indicated that there was a large proportion of taxa that remained undetected by our sampling regime (14-94% per site). Our sites were located across a gradient of agricultural activity, and while we predicted that increased land use would homogenize benthic communities, this was not supported as within-stream dissimilarity was unrelated to land use. Within-stream dissimilarity estimates were consistently high for all levels of taxonomic resolution (invertebrate families, invertebrate OTUs, chironomid OTUs), indicating stream communities are very dissimilar at small spatial scales.

Dataset Information

DNA metabarcoding assessment of Neotropical ichthyoplankton communities is marker-dependent.

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DNA metabarcoding assessment of Neotropical ichthyoplankton communities is marker-dependent.

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