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ABSTRACT In metabarcoding studies, Linnaean taxonomy assignments of Operational Taxonomic Units (OTUs) or Amplicon Sequence Variants (ASVs) underpin many downstream bioinformatics analyses and ecological interpretations of environmental DNA (eDNA) datasets. However, public molecular databases (i.e., SILVA, EUKARYOME, BOLD) for most microbial metazoan phyla (nematodes, tardigrades, kinorhynchs, etc.) are sparsely populated, negatively impacting our ability to assign ecologically meaningful taxonomy to these understudied groups. Additionally, the choice of bioinformatics parameters and computational algorithms can further affect the accuracy of eDNA taxonomy assignments. Here, we use twoin silicodatasets to show that taxonomy assignments using the 18S rRNA gene can be dramatically improved by curating Linnaean taxonomy strings associated with each reference sequence and closing phylogenetic gaps by improving taxon sampling. Using free‐living nematodes as a case study, we applied two commonly used taxonomy assignment algorithms (BLAST+ and the QIIME2 Naïve Bayes classifier) across six iterations of the SILVA 138 reference database to evaluate the precision and accuracy of taxonomy assignments. The BLAST+ top hit with a 90% sequence similarity cutoff often returned the highest percentage of correctly assigned taxonomy at the genus level, and the QIIME2 Naïve Bayes classifier performed similarly well when paired with a reference database containing corrected taxonomy strings. Our results highlight the urgent need for phylogenetically informed expansions of public reference databases (encompassing both genomes and common gene markers), focused on poorly sampled lineages that are now robustly recovered via eDNA metabarcoding approaches. Additional taxonomy curation efforts should be applied to popular reference databases such as SILVA, and taxon sampling could be rapidly improved by more frequent incorporation of newly published GenBank sequences linked to genus‐ and/or species‐level identifications.more » « lessFree, publicly-accessible full text available March 1, 2026
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Free, publicly-accessible full text available December 1, 2026
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Abstract The study of microbiomes across organisms and environments has become a prominent focus in molecular ecology. This perspective article explores common challenges, methodological advancements, and future directions in the field. Key research areas include understanding the drivers of microbiome community assembly, linking microbiome composition to host genetics, exploring microbial functions, transience and spatial partitioning, and disentangling non‐bacterial components of the microbiome. Methodological advancements, such as quantifying absolute abundances, sequencing complete genomes, and utilizing novel statistical approaches, are also useful tools for understanding complex microbial diversity patterns. Our aims are to encourage robust practices in microbiome studies and inspire researchers to explore the next frontier of this rapidly changing field.more » « less
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Abstract Environmental DNA (eDNA) data make it possible to measure and monitor biodiversity at unprecedented resolution and scale. As use‐cases multiply and scientific consensus grows regarding the value of eDNA analysis, public agencies have an opportunity to decide how and where eDNA data fit into their mandates. Within the United States, many federal and state agencies are individually using eDNA data in various applications and developing relevant scientific expertise. A national strategy for eDNA implementation would capitalize on recent scientific developments, providing a common set of next‐generation tools for natural resource management and public health protection. Such a strategy would avoid patchwork and possibly inconsistent guidelines in different agencies, smoothing the way for efficient uptake of eDNA data in management. Because eDNA analysis is already in widespread use in both ocean and freshwater settings, we focus here on applications in these environments. However, we foresee the broad adoption of eDNA analysis to meet many resource management issues across the nation because the same tools have immediate terrestrial and aerial applications.more » « less
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The CSSI 2019 workshop was held on October 28-29, 2019, in Austin, Texas. The main objectives of this workshop were to (1) understand the impact of the CSSI program on the community over the last 9 years, (2) engage workshop participants in identifying gaps and opportunities in the current CSSI landscape, (3) gather ideas on the cyberinfrastructure needs and expectations of the community with respect to the CSSI program, and (4) prepare a report summarizing the feedback gathered from the community that can inform the future solicitations of the CSSI program. The workshop participants included a diverse mix of researchers and practitioners from academia, industry, and national laboratories. The participants belonged to diverse domains such as quantum physics, computational biology, High Performance Computing (HPC), and library science. Almost 50% participants were from computer science domain and roughly 50% were from non-computer science domains. As per the self-reported statistics, roughly 27% of the participants were from the different underrepresented groups as defined by the National Science Foundation (NSF). The workshop brought together different stakeholders interested in provisioning sustainable cyberinfrastructure that can power discoveries impacting the various fields of science and technology and maintaining the nation's competitiveness in the areas such as scientific software, HPC, networking, cybersecurity, and data/information science. The workshop served as a venue for gathering the community-feedback on the current state of the CSSI program and its future directions. Before they arrived at the workshop, the participants were encouraged to take an online survey on the challenges that they face in using the current cyberinfrastructure and the importance of the CSSI program in enabling cutting-edge research. The workshop included 16 brain-storming sessions of one hour each. Additionally, the workshop program included 16 lightning talks and an extempore session. The information collected from the survey, brainstorming sessions, lightning talks, and the extempore session are summarized in this report and can potentially be useful for the NSF in formulating the future CSSI solicitations. The workshop fostered an environment in which the participants were encouraged to identify gaps and opportunities in the current cyberinfrastructure landscape, and develop thoughts for proposing new projects.more » « less
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Abstract Bloom‐forming gelatinous zooplankton occur circumglobally and significantly influence the structure of pelagic marine food webs and biogeochemical cycling through interactions with microbial communities. During bloom conditions especially, gelatinous zooplankton are keystone taxa that help determine the fate of primary production, nutrient remineralization, and carbon export. Using the pelagic tunicateDolioletta gegenbaurias a model system for gelatinous zooplankton, we carried out a laboratory‐based feeding experiment to investigate the potential ecosystem impacts of doliolid gut microbiomes and microbial communities associated with doliolid faecal pellets and the surrounding seawater. Metabarcoding targeting Bacteria and Archaea 16S rRNA genes/Archaea) and qPCR approaches were used to characterize microbiome assemblages. Comparison between sample types revealed distinct patterns in microbial diversity and biomass that were replicable across experiments. These observations support the hypothesis that through their presence and trophic activity, doliolids influence the structure of pelagic food webs and biogeochemical cycling in subtropical continental shelf systems where tunicate blooms are common. Bacteria associated with starved doliolids (representative of the resident gut microbiome) possessed distinct low‐biomass and low‐diversity microbial assemblages, suggesting that the doliolid microbiome is optimized to support a detrital trophic mode. Bacterial generaPseudoalteromomasandShimiawere the most abundant potential core microbiome taxa, similar to patterns observed in other marine invertebrates. Exploratory bioinformatic analyses of predicted functional genes suggest that doliolids, via their interactions with bacterial communities, may affect important biogeochemical processes including nitrogen, sulphur, and organic matter cycling.more » « less
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null (Ed.)Marine multicellular organisms host a diverse collection of bacteria, archaea, microbial eukaryotes, and viruses that form their microbiome. Such host-associated microbes can significantly influence the host’s physiological capacities; however, the identity and functional role(s) of key members of the microbiome (“core microbiome”) in most marine hosts coexisting in natural settings remain obscure. Also unclear is how dynamic interactions between hosts and the immense standing pool of microbial genetic variation will affect marine ecosystems’ capacity to adjust to environmental changes. Here, we argue that significantly advancing our understanding of how host-associated microbes shape marine hosts’ plastic and adaptive responses to environmental change requires (i) recognizing that individual host–microbe systems do not exist in an ecological or evolutionary vacuum and (ii) expanding the field toward long-term, multidisciplinary research on entire communities of hosts and microbes. Natural experiments, such as time-calibrated geological events associated with well-characterized environmental gradients, provide unique ecological and evolutionary contexts to address this challenge. We focus here particularly on mutualistic interactions between hosts and microbes, but note that many of the same lessons and approaches would apply to other types of interactions.more » « less
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