ABSTRACT
Plastic particles are ubiquitous in marine systems and fragment into smaller pieces, such as nanoplastics (NPs). The effects of NPs on marine organisms are of growing concern but are not well understood. Marine sediments act as a sink for many contaminants, like microplastics, and are rich habitats for benthic micro- and meiofauna which are ecologically-important components of marine food webs; however, little is known about the sensitivities of specific organisms to NPs or the effects on community diversity and composition. Utilizing molecular methods, such as metabarcoding of environmental DNA/RNA, allows for the rapid and comprehensive detection of microscopic organisms via high-throughput sequencing to assess adverse effects at the community level. The objective of this study was to use a metabarcoding approach to investigate the effects of NPs on benthic micro- and meiofaunal community diversity. Mesocosms were created with sediment cores collected from the Narrow River estuary (Rhode Island, USA) and exposed to 900 nm diameter weathered polystyrene beads at concentrations of 0.1, 1, 10, or 100 mg/kg dry weight in sediment for two weeks. Following exposure, RNA and DNA were co-extracted from the sediment, RNA was reverse-transcribed, 18S and COI markers were PCR-amplified, and amplicons were sequenced on an Illumina MiSeq. Using the 18S marker and eRNA template, increases to α-diversity and significant differences to ß-diversity were observed in the highest NP exposures relative to the control. Observed differences in community composition were driven by the differential abundance of several types of protists and arthropods. Significant dose-dependent shifts in composition were observed in ß-diversity Jaccard and Unweighted-Unifrac metrics with the 18S marker using the RNA template. To our knowledge, this is the first demonstration of a dose-response relationship for NPs at a community level, and it highlights the value of using community-level endpoints to assess environmental impacts of nanoparticles.
Subject(s)
DNA, Environmental , Ecosystem , Microplastics , Biodiversity , Plastics/toxicity , DNA Barcoding, Taxonomic , RNAABSTRACT
Achieving gender equity is a long-standing and ubiquitous challenge in marine science. Creating equitable experiences for all genders in marine science requires recognizing scientists' intersectional identities, and how this leads to unique lived experiences of privilege and marginalization. One approach to increase equitable experiences for women in marine science is to create affinity groups where women can learn from each other, share their experiences, and provide support and mentorship. The Society for Women in Marine Science (SWMS) is one such organization, founded to amplify the work of early career women in marine science and create community, through events such as full-day symposium events. This study investigates the experiences of symposium attendees for four events held from 2018 through 2020, as reported in pre- and post-symposium surveys. We used quantitative analysis of the open-ended survey questions to examine the demographics of attendees and their fields of study. Qualitative thematic analysis identified the most effective aspects of the symposia, areas of logistical and content improvement for future symposia, and emphasized the unique challenges women in marine science experience. The majority of symposium attendees were white graduate students. Nearly all attendees identified as women, with a small number of men and non-binary individuals. Symposia attendees enjoyed opportunities for professional development and interactions with colleagues across career stages. We present recommendations for continuing to foster a sense of belonging in marine science and STEM more broadly, both specific to SWMS and transferable actions that can be applied for other affinity groups. These suggestions include empathetic event logistics, continual democratic evaluation, identity reflexivity among group leaders, and professional development activities targeted towards the unique needs of the affinity group. The positive responses received from SWMS's adaptive integration of survey results into symposia demonstrate that incorporating these recommendations and findings will help create an inclusive wave in marine science.
ABSTRACT
Microscopic organisms are often overlooked in traditional diversity assessments due to the difficulty of identifying them based on morphology. Metabarcoding is a method for rapidly identifying organisms where Environmental DNA (eDNA) is used as a template. However, legacy DNA is problematically detected from organisms no longer in the environment during sampling. Environmental RNA (eRNA), which is only produced by living organisms, can also be collected from environmental samples and used for metabarcoding. The aim of this study was to determine differences in community composition and diversity between eRNA and eDNA templates for metabarcoding. Using mesocosms containing field-collected communities from an estuary, RNA and DNA were co-extracted from sediment, libraries were prepared for two loci (18S and COI), and sequenced using an Illumina MiSeq. Results show a higher number of unique sequences detected from eRNA in both markers and higher α-diversity compared to eDNA. Significant differences between eRNA and eDNA for all ß-diversity metrics were also detected. This study is the first to demonstrate community differences detected with eRNA compared to eDNA from an estuarine system and illustrates the broad applications of eRNA as a tool for assessing benthic community diversity, particularly for environmental conservation and management applications.
Subject(s)
DNA, Environmental , DNA, Environmental/genetics , DNA Barcoding, Taxonomic/methods , RNA/genetics , Environmental Monitoring/methods , Biodiversity , DNA/geneticsABSTRACT
Several populations of the amphipod, Hyalella azteca, have developed resistance to pyrethroid insecticides due to non-target exposure, but the dominance of the resistance trait is unknown. The current study investigated the dominance level of point mutations in natural populations of insecticide-resistant H. azteca and determined whether H. azteca from different clades with and without resistant alleles can hybridize and produce viable offspring. A parent generation (P0) of non-resistant homozygous wild type H. azteca was crossbred with pyrethroid-resistant homozygous mutant animals and the tolerance of the filial 1 (F1) generation to the pyrethroid insecticide, permethrin, was measured. Then the genotypes of the F1 generation was examined to assure heterozygosity. The resistant parents had permethrin LC50 values that ranged from 52 to 82 times higher than the non-resistant animals and both crossbreeding experiments produced heterozygous hybrid offspring that had LC50 values similar to the non-resistant H. azteca parent. Dominance levels calculated for each of the crosses showed values close to 0, confirming that the L925I and L925V mutations were completely recessive. The lack of reproduction by hybrids of the C x D breeding confirmed that these clades are reproductively isolated and therefore introgression of adaptive alleles across these clades is unlikely. Potential evolutionary consequences of this selection include development of population bottlenecks, which may arise leading to fitness costs and reduced genetic diversity of H. azteca.