Quantitative assessment of visual microscopy as a tool for microplastic research: Recommendations for improving methods and reporting.

Microscopy is often the first step in microplastic analysis and is generally followed by spectroscopy to confirm material type. The value of microscopy lies in its ability to provide count, size, color, and morphological information to inform toxicity and source apportionment. To assess the accuracy and precision of microscopy, we conducted a method evaluation study. Twenty-two laboratories from six countries were provided three blind spiked clean water samples and asked to follow a standard operating procedure. The samples contained a known number of microplastics with different morphologies (fiber, fragment, sphere), colors (clear, white, green, blue, red, and orange), polymer types (PE, PS, PVC, and PET), and sizes (ranging from roughly 3-2000 µm), and natural materials (natural hair, fibers, and shells; 100-7000 µm) that could be mistaken for microplastics (i.e., false positives). Particle recovery was poor for the smallest size fraction (3-20 µm). Average recovery (±StDev) for all reported particles >50 µm was 94.5 ± 56.3%. After quality checks, recovery for >50 µm spiked particles was 51.3 ± 21.7%. Recovery varied based on morphology and color, with poorest recovery for fibers and the largest deviations for clear and white particles. Experience mattered; less experienced laboratories tended to report higher concentration and had a higher variance among replicates. Participants identified opportunity for increased accuracy and precision through training, improved color and morphology keys, and method alterations relevant to size fractionation. The resulting data informs future work, constraining and highlighting the value of microscopy for microplastics.

Sampling and Quality Assurance and Quality Control: A Guide for Scientists Investigating the Occurrence of Microplastics Across Matrices.

Plastic pollution is a defining environmental contaminant and is considered to be one of the greatest environmental threats of the Anthropocene, with its presence documented across aquatic and terrestrial ecosystems. The majority of this plastic debris falls into the micro (1 µm-5 mm) or nano (1-1000 nm) size range and comes from primary and secondary sources. Its small size makes it cumbersome to isolate and analyze reproducibly, and its ubiquitous distribution creates numerous challenges when controlling for background contamination across matrices (e.g., sediment, tissue, water, air). Although research on microplastics represents a relatively nascent subfield, burgeoning interest in questions surrounding the fate and effects of these debris items creates a pressing need for harmonized sampling protocols and quality control approaches. For results across laboratories to be reproducible and comparable, it is imperative that guidelines based on vetted protocols be readily available to research groups, many of which are either new to plastics research or, as with any new subfield, have arrived at current approaches through a process of trial-and-error rather than in consultation with the greater scientific community. The goals of this manuscript are to (i) outline the steps necessary to conduct general as well as matrix-specific quality assurance and quality control based on sample type and associated constraints, (ii) briefly review current findings across matrices, and (iii) provide guidance for the design of sampling regimes. Specific attention is paid to the source of microplastic pollution as well as the pathway by which contamination occurs, with details provided regarding each step in the process from generating appropriate questions to sampling design and collection.

Reporting Guidelines to Increase the Reproducibility and Comparability of Research on Microplastics.

The ubiquitous pollution of the environment with microplastics, a diverse suite of contaminants, is of growing concern for science and currently receives considerable public, political, and academic attention. The potential impact of microplastics in the environment has prompted a great deal of research in recent years. Many diverse methods have been developed to answer different questions about microplastic pollution, from sources, transport, and fate in the environment, and about effects on humans and wildlife. These methods are often insufficiently described, making studies neither comparable nor reproducible. The proliferation of new microplastic investigations and cross-study syntheses to answer larger scale questions are hampered. This diverse group of 23 researchers think these issues can begin to be overcome through the adoption of a set of reporting guidelines. This collaboration was created using an open science framework that we detail for future use. Here, we suggest harmonized reporting guidelines for microplastic studies in environmental and laboratory settings through all steps of a typical study, including best practices for reporting materials, quality assurance/quality control, data, field sampling, sample preparation, microplastic identification, microplastic categorization, microplastic quantification, and considerations for toxicology studies. We developed three easy to use documents, a detailed document, a checklist, and a mind map, that can be used to reference the reporting guidelines quickly. We intend that these reporting guidelines support the annotation, dissemination, interpretation, reviewing, and synthesis of microplastic research. Through open access licensing (CC BY 4.0), these documents aim to increase the validity, reproducibility, and comparability of studies in this field for the benefit of the global community.

Microplastics Differ Between Indoor and Outdoor Air Masses: Insights from Multiple Microscopy Methodologies.

The abundance and distribution of microplastic (<5 mm) has become a growing concern, particularly over the past decade. Research to date has focused on water, soil, and organism matrices but generally disregarded air. We explored airborne microplastic inside and outside of buildings in coastal California by filtering known volumes of air through glass fiber filters, which were then subsequently characterized with a variety of microscopy techniques: gross traditional microscopy, fluorescent microscopy following staining with Nile red, micro-Raman spectroscopy, and micro-Fourier transform infrared (µFT-IR) spectroscopy. Microplastics permeated the air, with indoor (3.3 ± 2.9 fibers and 12.6 ± 8.0 fragments m-3; mean ± 1 SD) harboring twice as much as outdoor air (0.6 ± 0.6 fibers and 5.6 ± 3.2 fragments m-3). Microplastic fiber length did not differ significantly between indoor and outdoor air, but indoor microplastic fragments (58.6 ± 55 µm) were half the size of outdoor fragments (104.8 ± 64.9 µm). Micro-Raman and FT-IR painted slightly different pictures of airborne plastic compounds, with micro-Raman suggesting polyvinyl chloride dominates indoor air, followed by polyethylene (PE) and µFT-IR showing polystyrene dominates followed by PE and polyethylene terephthalate. The ubiquity of airborne microplastic points to significant new potential sources of plastic inputs to terrestrial and marine ecosystems and raises significant concerns about inhalation exposure to humans both indoors and outdoors.

Microplastics are ubiquitous on California beaches and enter the coastal food web through consumption by Pacific mole crabs.

Microplastics are commonly found in marine ecosystems, but their distribution, prevalence, and impacts on resident fauna are still not well understood. Microplastics in coastal sediments expose invertebrate infauna to the risk of ingestion of plastic debris and associated toxicants. We assessed the prevalence of microplastics in beach sediments and ingested by Pacific mole crabs (Emerita analoga) at sandy beaches spanning >900 km of the California coast. Microplastics were present in sediments of every one of 51 beaches sampled. At a subset of 38 beaches Pacific mole crabs were collected and crabs at every beach had ingested microplastics. Across all beaches sampled, an average of 35% of Pacific mole crabs examined had microplastics in their guts. Our study demonstrates that microplastics are ubiquitous in sediments on California beaches and they are frequently consumed by a filter-feeding crustacean that is a common prey item in the diet of a wide variety of taxa, including fishes and birds.