Genetic taxonomic assignment can be more sensitive than morphological taxonomic assignment, particularly for small, cryptic or rare species. Sequence processing is essential to taxonomic assignment, but can also produce errors because optimal parameters are not known a priori. Here, we explored how sequence processing parameters influence taxonomic assignment of 18S sequences from bulk zooplankton samples produced by 454 pyrosequencing. We optimized a sequence processing pipeline for two common research goals, estimation of species richness and early detection of aquatic invasive species (AIS), and then tested most optimal models’ performances through simulations. We tested 1,050 parameter sets on 18S sequences from 20 AIS to determine optimal parameters for each research goal. We tested optimized pipelines’ performances (detectability and sensitivity) by computationally inoculating sequences of 20 AIS into ten bulk zooplankton samples from ports across Canada. We found that optimal parameter selection generally depends on the research goal. However, regardless of research goal, we found that metazoan 18S sequences produced by 454 pyrosequencing should be trimmed to 375–400 bp and sequence quality filtering should be relaxed (1.5 ≤ maximum expected error ≤ 3.0, Phred score = 10). Clustering and denoising were only viable for estimating species richness, because these processing steps made some species undetectable at low sequence abundances which would not be useful for early detection of AIS. With parameter sets optimized for early detection of AIS, 90% of AIS were detected with fewer than 11 target sequences, regardless of whether clustering or denoising was used. Despite developments in next‐generation sequencing, sequence processing remains an important issue owing to difficulties in balancing false‐positive and false‐negative errors in metabarcoding data.
The ecological impacts of invasive species are highly variable and mediated by many factors, including both habitat and population abundance. Lionfish Pterois volitans are an invasive marine species which have high reported detrimental effects on prey populations, but whose effects relative to native predators are currently unknown for the recently colonized eastern Gulf of Mexico. We used functional response (FR) methodology to assess the ecological impact of lionfish relative to 2 functionally similar native species (red grouper Epinephelus morio and graysby grouper Cephalopholis cruentata) foraging in a heterogeneous environment. We then combined the per capita impact of each species with their field abundance to obtain a Relative Impact Potential (RIP). RIP assesses the broader ecological impact of invasive relative to native predators, the magnitude of which predicts community-level negative effects of invasive species. Lionfish FR and overall consumption rate was intermediate to that of red grouper (higher) and graysby grouper (lower). However, lionfish had the highest capture efficiency of all species, which was invariant of habitat. Much higher field abundance of lionfish resulted in high RIPs relative to both grouper species, demonstrating that the ecological impact of lionfish in this region will be driven mainly by high abundance and high predator efficiency rather than per capita effect. Our comparative study is the first empirical assessment of lionfish per capita impact and RIP in this region and is one of few such studies to quantify the FR of a marine predator.
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