How many microplastics do you need to (sub)sample?

Analysis of microplastics in the environment requires polymer characterization as a confirmation step for suspected microplastic particles found in a sample. Material characterization is costly and can take a long time per particle. When microplastic particle counts are high, many researchers cannot characterize every particle in their sample due to time or monetary constraints. Moreover, characterizing every particle in samples with high plastic particle counts is unnecessary for describing the sample properties. We propose an a priori approach to determine the number of suspected microplastic particles in a sample that should be randomly subsampled for characterization to accurately assess the polymer distribution in the environmental sample. The proposed equation is well-founded in statistics literature and was validated using published microplastic data and simulations for typical microplastic subsampling routines. We report values from the whole equation but also derive a simple way to calculate the necessary particle count for samples or subsamples by taking the error to the power of negative two. Assuming an error of 0.05 (5 %) with a confidence interval of 95 %, an unknown expected proportion, and a sample with many particles (> 100k), the minimum number of particles in a subsample should be 386 particles to accurately characterize the polymer distribution of the sample, given the particles are randomly characterized from the full population of suspected particles. Extending this equation to simultaneously estimate polymer, color, size, and morphology distributions reveals more particles (620) would be needed in the subsample to achieve the same high absolute error threshold for all properties. The above proposal for minimum subsample size also applies to the minimum count that should be present in samples to accurately characterize particle type presence and diversity in a given environmental compartment.

Steps Scientists Can Take to Inform Aquatic Microplastics Management: A Perspective Informed by the California Experience.

Recent evidence suggests that microplastic particles are pervasive and potentially of great risk to both animal and human health. The California legislature has responded to this information by enacting two new bills that require quantification of microplastics in various media and development of new management strategies to address microplastic pollution. However, there are several scientific gaps that impede the development and implementation of necessary management strategies to address microplastic pollution. In this paper, we use the California experience as a case study to provide perspective on those science gaps, the current barriers to science affecting management, and the actions scientists can take to best ensure their efforts are of greatest value to policymakers and the management community.

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.

Ecotoxicogenomics: Microarray interlaboratory comparability.

Transcriptomic analysis can complement traditional ecotoxicology data by providing mechanistic insight, and by identifying sub-lethal organismal responses and contaminant classes underlying observed toxicity. Before transcriptomic information can be used in monitoring and risk assessment, it is necessary to determine its reproducibility and detect key steps impacting the reliable identification of differentially expressed genes. A custom 15K-probe microarray was used to conduct transcriptomics analyses across six laboratories with estuarine amphipods exposed to cyfluthrin-spiked or control sediments (10 days). Two sample types were generated, one consisted of total RNA extracts (Ex) from exposed and control samples (extracted by one laboratory) and the other consisted of exposed and control whole body amphipods (WB) from which each laboratory extracted RNA. Our findings indicate that gene expression microarray results are repeatable. Differentially expressed data had a higher degree of repeatability across all laboratories in samples with similar RNA quality (Ex) when compared to WB samples with more variable RNA quality. Despite such variability a subset of genes were consistently identified as differentially expressed across all laboratories and sample types. We found that the differences among the individual laboratory results can be attributed to several factors including RNA quality and technical expertise, but the overall results can be improved by following consistent protocols and with appropriate training.

Fish consumption as a driver of risk-management decisions and human health-based water quality criteria.

The use and interpretation of fish consumption surveys and interviews, the application of fish consumption rates for sediment evaluation and cleanup, and the development of human health water quality criteria (HH WQC) are complex and interrelated issues. The present article focuses on these issues using examples from the United States, although the issues may be relevant for other countries. Some key considerations include the fact that there are many types of fish consumption surveys (e.g., 24-h recall surveys, food frequency questionnaires, creel surveys), and these surveys have different advantages and limitations. Identification of target populations for protection, identification of the species and quantities of fish consumed, and determination of bioaccumulation assumptions are important factors when developing water quality and sediment screening levels and standards. Accounting for the cultural importance of fish consumption for some populations is an even more complex element. Discussions about HH WQC often focus only on the fish consumption rate and may not have broad public input. Some states are trying to change this through extensive public participation efforts and use of probabilistic approaches to derive HH WQC. Finally, there are limits to what WQC can achieve. Solutions beyond the establishment of WQC that target toxics reduction from other sources may provide the greatest improvements to water quality and reductions in human health risks in the future.

Plastic ingestion by planktivorous fishes in the North Pacific Central Gyre.

A significant amount of marine debris has accumulated in the North Pacific Central Gyre (NPCG). The effects on larger marine organisms have been documented through cases of entanglement and ingestion; however, little is known about the effects on lower trophic level marine organisms. This study is the first to document ingestion and quantify the amount of plastic found in the gut of common planktivorous fish in the NPCG. From February 11 to 14, 2008, 11 neuston samples were collected by manta trawl in the NPCG. Plastic from each trawl and fish stomach was counted and weighed and categorized by type, size class and color. Approximately 35% of the fish studied had ingested plastic, averaging 2.1 pieces per fish. Additional studies are needed to determine the residence time of ingested plastics and their effects on fish health and the food chain implications.

Association of ALDH1 promoter polymorphisms with alcohol-related phenotypes in Trinidad and Tobago.

OBJECTIVE: Two polymorphisms in the promoter region of the gene encoding cytosolic aldehyde dehydrogenase (ALDH1A1), ALDH1A1*2 and ALDH1A1*3, have recently been identified. The present study sought to determine whether an association exists between ALDH1A1 genotypes, alcohol dependence, drinking history, and liver function tests in the two major ethnic groups of Trinidad and Tobago (TT). METHOD: The participants in this study were 137 alcohol dependents of either East Indian ancestry (Indo-TT) or African ancestry (Afro-TT) and 108 controls matched by age, gender, and ethnicity. A structured interview was used to gather information on demographics, psychiatric diagnoses, and personal drinking and drug use. A blood sample was obtained from each participant, and leukocyte DNA was extracted and used to genotype for the presence of the ALDH1A1 promoter polymorphisms. Serum levels of hepatic enzymes, as well as presence of HIV, hepatitis B surface antigen, and antihepatitis C virus antibody, were also determined. RESULTS: Twenty-four participants (10%) possessed the ALDH1A1*1/*2 genotype (frequency = .05), 4 were Afro-TT (2 alcohol dependents, 2 controls), and 20 were Indo-TT (18 alcohol dependents, 2 controls). Two participants (1 Indo-TT alcohol dependent, 1 Afro-TT alcohol dependent) had the ALDH1A1*2/*2 genotype. Four participants possessed ALDH1A1*3, all of whom were Afro-TT controls. Indo-TT participants with at least one ALDH1A1*2 allele were more likely to have a lifetime diagnosis of Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised, alcohol dependence (p < .002). Indo-TT participants with ALDH1A1*2 also reported significantly higher levels of current alcohol consumption (p < .05). The small number of Afro-TT participants with atypical polymorphisms limits any conclusions on the possible impact on alcohol dependence in that population. CONCLUSIONS: Results from this study suggest that ALDH1A1*2 may be associated with increased risk for the development of alcohol dependence in Indo-Trinidadians.

Association of the ADHIB*3 allele with alcohol-related phenotypes in Trinidad.

BACKGROUND: Two of the class I alcohol dehydrogenase (ADH) genes located on chromosome 4 (ADH1B and ADH1C) encode for multiple isozymes that differ in their kinetic properties. At the ADH1B locus, 3 polymorphisms are present (ADH1B(*)1, ADH1B(*)2, ADH1B(*)3). ADH1B(*)2 (found mostly in individuals of East Asian and Jewish descent) and ADH1B(*)3 (found mostly in individuals of African decent) alleles encode for a more active enzyme variants than ADH1B(*)1 and the presence of these alleles has been associated with protection from alcohol dependence. The relationship between these alleles and alcohol-associated phenotypes has not been previously investigated in individuals living in the Caribbean. METHODS: One hundred thirty-three alcohol-dependent individuals of either East Indian or African ancestry and 98 controls matched by age, sex, education, and ethnicity participated in the study. A structured interview [the Semi-Structured Assessment for the Genetics of Alcoholism (SSAGA)] was used to gather information on demographics, psychiatric diagnoses, personal drinking, and drug use history. Leukocyte DNA extracted from a blood sample obtained from each participant was genotyped at the ADH1B locus. Serum levels of the liver enzymes alanine and aspartate aminotransferase (ALT, AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), and gamma-glutamyl transferase (GGT) as well as the presence of HIV, hepatitis B surface antigen, and antihepatitis C virus antibody were also assayed. The specific aim of the study was to investigate the associations between ADH1B alleles and alcohol dependence, drinking history, and liver function in individuals from the 2 major ethnic groups of Trinidad (individuals of African and East Indian ancestry). RESULTS: Twenty-eight of the Afro-Trinidadian (Afro-TT) participants (41%) and 1 Indo-Trinidadian (Indo-TT) (>1%) had at least 1 ADH1B(*)3 allele and 3 Afro-TT were homozygous for the allele. African participants with at least 1 ADH1B(*)3 allele were found to be significantly less likely to be alcohol dependent (p<0.018), and to have lower alcohol consumption levels (p<0.05). Among those participants who were alcohol dependent, ADH1B(*)3 was associated with significantly higher levels of ALT (p<0.05). CONCLUSIONS: This study suggests, in this sample of Trinidadians, that the ADH1B(*)3 allele is associated with protection from the development of alcoholism but is also associated with enhanced risk for elevated serum ALT levels in those individuals who do become alcohol dependent.