Taking control of microplastics data: A comparison of control and blank data correction methods

Although significant headway has been achieved regarding method harmonisation for the analysis of microplastics, analysis and interpretation of control data has largely been overlooked. There is currently no consensus on the best method to utilise data generated from controls, and consequently many methods are arbitrarily employed. This study identified 6 commonly implemented strategies: a) No correction; b) Subtraction; c) Mean Subtraction; d) Spectral Similarity; e) Limits of detection/ limits of quantification (LOD/LOQ) or f) Statistical analysis, of which many variations are possible. Here, the 6 core methods and 45 variant methods (n = 51) thereof were used to correct a dummy dataset using control data. Most of the methods tested were too inflexible to account for the inherent variation present in microplastic data. Only 7 of the 51 methods tested (six LOD/LOQ methods and one statistical method) showed promise, removing between 96.3 % and 100 % of the contamination data from the dummy set. The remaining 44 methods resulted in deficient corrections for background contamination due to the heterogeneity of microplastics. These methods should be avoided in the future to avoid skewed results, especially in low abundance samples. Overall, LOD/LOQ methods or statistical analysis comparing means are recommended for future use in microplastic studies

Ingestion and depuration of microplastics by a planktivorous coral reef fish, Pomacentrus amboinensis

Microplastics are ubiquitous contaminants in marine environments and organisms. Concerns about potential impacts on marine organisms are usually associated with uptake of microplastics, especially via ingestion. This study used environmentally relevant exposure conditions to investigate microplastic ingestion and depuration kinetics of the planktivorous damselfish, Pomacentrus amboinensis. Irregular shaped blue polypropylene (PP) particles (longest length 125–250 μm), and regular shaped blue polyester (PET) fibers (length 600–700 μm) were selected based on physical and chemical characteristics of microplastics commonly reported in the marine environment, including in coral reef ecosystems. Individual adult damselfish were exposed to a single dose of PP particles and PET fibers at concentrations reported for waters of the Great Barrier Reef (i.e., environmentally relevant concentrations, ERC), or future projected higher concentrations (10x ERC, 100x ERC). Measured microplastic concentrations were similar to their nominal values, confirming that PP particles and PET fibers were present at the desired concentrations and available for ingestion by individual damselfish. Throughout the 128-h depuration period, the 88 experimental fish were sampled 2, 4, 8, 16, 32, 64, and 128-h post microplastic exposure and their gastrointestinal tracts (GIT) analyzed for ingested microplastics. While damselfish ingested both experimental microplastics at all concentrations, body burden, and depuration rates of PET fibers were significantly larger and longer, respectively, compared to PP particles. For both microplastic types, exposure to higher concentrations led to an increase in body burden and lower depuration rates. These findings confirm ingestion of PP particles and PET fibers by P. amboinensis and demonstrate for the first time the influence of microplastic characteristics and concentrations on body burden and depuration rates. Finally, despite measures put in place to prevent contamination, extraneous microplastics were recovered from experimental fish, highlighting the challenge to completely eliminate contamination in microplastic exposure studies. These results are critical to inform and continuously improve protocols for future microplastics research, and to elucidate patterns of microplastic contamination and associated risks in marine organisms

Relevance and reliability of evidence for microplastic contamination in seafood: A critical review using Australian consumption patterns as a case study

Seafood contamination with, and human consumption of, microplastics (MPs) have recently been highlighted as an emerging concern for global food security. While there is evidence that commercial marine species are contaminated with MPs, it is still unknown if seafood can act as a vector for MP transfer to human consumers. Microplastics have been reported in the digestive tract, gills and in select internal organs of marine animals. However, many of these tissues are not typically eaten by human consumers but discarded. In this critical review, we examined the peer-reviewed literature for evidence of MP contamination in seafood, and the potential transfer to human consumers. Based on known seafood consumption patterns in a typical Australian diet, we assessed the relevance and reliability of the current body of literature to examine the prospect and risk of MP transfer. The relevance of data was considered based on the organism studied, origin of the samples, and the tissues analysed, while reliability was assessed based on procedural methodologies used to derive the data. A review of 132 studies found limited evidence of MP contamination in edible tissues from fresh fish or crustaceans. MP presence was confirmed in packaged fish, as well as in fresh and packaged bivalve molluscs. The limited number of studies satisfying the relevance and reliability criteria (n = 24) precluded a quantitative assessment of the potential risk associated with MP transfer. While consumption of packaged fish and bivalve molluscs may result in the consumption of MPs by humans, it is currently unknown whether this presents a health risk

An assessment workflow to recover microplastics from complex biological matrices

A criteria-guided workflow was applied to assess the effectiveness of microplastic separation methods on complex marine biological matrices. Efficacy of four methods (nitric acid, HNO3, and potassium hydroxide, KOH, digestions, and sodium chloride, NaCl, and potassium iodide, KI, density flotations) was evaluated on four taxa (hard coral, sponge, sea squirt, sea cucumber) using five microplastics (polyethylene, polystyrene, polyethylene terephthalate, PET, polyvinylchloride, rayon). Matrix clarification was only unacceptably low for KOH. PET discoloured regardless of reagent. Rayon threads unravelled into monofilaments after exposure to all reagents, with discolouration also occurring with HNO3. Recovery rates were overall high, except for dense microplastics treated with NaCl and only KI yielded high rayon recovery efficiency. All polymers were accurately assigned, with subtle spectral changes observed. These results demonstrate specific limitations to separation methods applied to different biological matrices and microplastics and highlight the need to assess their suitability to provide estimates of microplastic contamination

Continuous exposure to microplastics does not cause physiological effects in the cultivated mussel Perna perna

The environmental impact of microplastics is a challenging theme, especially under realistic experimental conditions. We investigated physiological responses to 0.1-1.0 mu m PVC particles intake by the mussel Perna perna after a relative long-term exposure (90 days) at a less extreme concentration compared with previous studies (0.125 g/L). Microplastic intake was inferred by the presence of PVC in the feces of mussels, and physiological damages were assessed through ingestion rate, assimilation efficiency, growth rate, cellular and molecular biomarkers (lysosomal integrity, lipid peroxidation, and DNA damage), and condition index. All physiological responses showed nonsignificant effects of the microplastics on the exposed mussels. We suggest that, despite the experimental concentration of microplastics, mussels were able to acclimate to the exposure through their abilities for long-term recovery and tolerance to stresses. These data have positive implications for environmental health and in terms of human food resource because mussel farming is a worldwide practice that heavily relies on plastic materials, increasing the chances of microplastic exposure and mussels contamination

Temporal patterns of plastic contamination in surface waters at the SS Yongala shipwreck, Great Barrier Reef, Australia

Plastic pollution is ubiquitous within the marine environment, including surface waters, water column and benthic sediments. Marine plastic contamination is expected to increase if future projections of increased plastic production eventuate. Conversely, national and international efforts are aiming to reduce marine plastic contamination. In this context, scientists, managers and the general public are increasingly interested in understanding the status and temporal trends of plastic contamination in the marine environment. Presented here is the first temporal assessment of plastic contamination in surface waters of the Great Barrier Reef (GBR), Australia. Specifically, duplicate surface seawater samples (n = 66) were collected at the SS Yongala shipwreck (Central GBR) monthly from September 2016 to September 2019 and analysed for plastic presence and abundance. The processing workflow involved density separation, followed by filtration, visual identification and sizing of putative plastics using stereomicroscopy, and chemical characterisation using Fourier transform infrared spectroscopy. A total of 533 plastic items were identified across all tows, consisting of macro-, meso- and microplastic fragments and fibres, with polypropylene and polyethylene being the most common polymers. Plastic contamination was detected in every replicate tow, bar one. Plastic concentrations fluctuated and spiked every three months, although contamination did not significantly alter across the three-year period. Wind speed, salinity and river discharge volume, but not surface current speed nor sea surface temperature, had a significant influence on the levels of plastic contamination. This study reveals, for the first time, the chronic presence of plastic debris in the surface waters of the GBR highlighting the need for long-term and on-going monitoring of the marine environment for plastic contamination

Improved microplastic processing from complex biological samples using a customized vacuum filtration apparatus

Plastics represent the largest component of marine debris globally. In this context, it is essential to quantify the current extent of plastic pollution, including microplastics (MP; plastics < 5 mm), within marine abiotic and biotic compartments. Despite significant effort, MP studies still face methodological impediments to establish accurate and standardized protocols to separate, process and analyze MPs in environmental samples. Furthermore, underestimation and overestimation of MP contamination, either through loss of MPs or introduction of extraneous MPs during handling and processing, is concerning, particularly when assessing risk profiles for marine ecosystems. Presented here is a custom-made stainless steel vacuum filtration apparatus designed to perform size-tiered separation and facilitate retrieval of MPs from a variety of environmental sample matrices. Incorporating this apparatus into a standard MP workflow achieved efficient graduated separation of commonly found MP fragments and fibers, validated by spike-recovery tests. As a case study, the gastrointestinal tracts of three juvenile Australian sharpnose sharks, Rhizoprionodon taylori, were processed using the filtration apparatus, and 46 anthropogenic items ranging from 0.021 to 8.87 mm were retrieved. This study demonstrates the effective use of the size-tiered stainless steel vacuum filtration apparatus and an improved efficiency in downstream microphotography and spectroscopic analyses of MPs from a complex sample matrix. Finally, it contributes to the MP research field by delivering more reliable estimates of MP contamination in marine ecosystems