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Sensitivity and accuracy of high-throughput metabarcoding methods used to describe aquatic communities for early detection of invasve fish species
Citation:
Hatzenbuhler, C., J. Hoffman, J. Kelly, J. Lietz, J. Martinson, S. Okum, Greg Peterson, AND E. Pilgrim. Sensitivity and accuracy of high-throughput metabarcoding methods used to describe aquatic communities for early detection of invasve fish species. Society of Freshwater Science, Sacramento, CA, May 20 - 26, 2016.
Impact/Purpose:
not applicable
Description:
For early detection biomonitoring of aquatic invasive species, sensitivity to rare individuals and accurate, high-resolution taxonomic classification are critical to minimize Type I and II detection errors. Given the great expense and effort associated with morphological identification of many aquatic organisms, high-throughput metabarcoding has gained recognition as an alternative means to describe biodiversity in aquatic communities. Our research uses complementary lab- and field- based experiments to investigate sensitivity and accuracy of metabarcoding methods commonly used to characterize sample composition (specifically, larval fish assemblages). Lab experiments demonstrate pyrosequencing of genetic marker CO1 (cytochrome c oxidase) facilitates detection of fish species with biomass percentages as low as 0.02-1%of total sample mass and results from parallel morphological and genetic analyses of field samples demonstrate metabarcoding can improve species richness estimates. Limits to detection, however, vary interspecifically and are influenced by CO1 amplification bias and data processing methods that skew sequence biodiversity estimates from corresponding constructed biodiversity and increase the rate of detection errors. Our findings directed field sample and sequence data processing method modifications aimed at reducing bias and ultimately detection error.