Identifying laboratory sources of microplastic and nanoplastic contamination from the air, water, and consumables.

Microplastic and nanoplastic research has proliferated in recent years in response to the escalating plastic pollution crisis. However, a lack of optimised methods for sampling and sample processing has potential implications for contaminating samples resulting in an overestimation of the quantity of microplastics and nanoplastics present in environmental samples. In response, a series of recommendations have been made, but most have not been quantified or validated sources of contamination. In the present study, we investigated sources of plastic contamination in common laboratory procedures including water sources (e.g., Milli-Q), consumables (e.g., unburnt glassware), airflow (e.g., fume hood) and dust. Using flow cytometry, we identified water, air flow and dust as sources of significant contamination. Milli-Q and reverse osmosis were the least contaminated sources when compared with tap water. Interestingly, current recommendations are to use glass consumables in replacement of plastic consumables, however, we have identified glassware and glass consumables as a significant source of contamination. Current best practice is to cover the glass tube with aluminium foil to reduce airborne contamination, but we found fresh aluminium foil to be a significant source of contamination, bringing light to the limitations foil has as a contamination control measure. Lastly, we identified significant quantities of microplastics and nanoplastics present in dust collected within the laboratory, suggesting this is a widespread and underestimated source of contamination. We have provided validated sources of contamination for both consumables and common laboratory procedures and provided mitigation strategies based on these. Additional recommendations include the appropriate design of experimental controls to quantify levels of introduced contamination based on methods and the detection techniques utilised. The application of these mitigation strategies and appropriate experimental design will allow for more accurate estimations on the level of microplastic and nanoplastic contamination within environmental samples.

The understudied global experiment of pollution's impacts on wildlife and human health: The ethical imperative for interdisciplinary research.

The global impact of pollution on human and wildlife health is a growing concern. The health impacts of pollution are significant and far-reaching yet poorly understood as no one field of research has the practices and methodologies required to encapsulate the diversity of these consequences. This paper advocates that interdisciplinary research is essential to comprehend the full extent of the impact of pollution. Medical and ecological research play a key role in investigating the health consequences of the pollution crisis, yet the wildlife experience is often neglected. This paper outlines how applying advanced techniques and expertise adapted in medical research to wildlife exposed to pollutants offers a unique perspective to understanding the full diversity of impacts to health. The challenges that impede the progress of this research include the lack of support for interdisciplinary research among funding streams, limitations in field-specific techniques, and a lack of communication between researchers from different disciplines. Of awarded funding from major national research councils across Australia, Europe, and the United States of America, only 0.5% is dedicated to pollution focused research. This is inclusive of laboratory equipment, mitigation strategies, quantification of environmental samples and health consequences research. Of that, 0.03% of funding is awarded to explaining the wildlife experience and documenting the health consequences observed despite being model organisms to environmentally and biologically relevant models for pollution exposure. This calls for a coordinated effort to overcome these hurdles and to promote interdisciplinary research in order to fully comprehend the consequences of pollution exposure and protect the health of humans, wildlife, and the environment. An interdisciplinary approach to this problem is timely given the magnitude of negative health consequences associated with exposure, the number of pollutants already present within the environment and the continual development of new compounds.

Assessing the effectiveness of MARPOL Annex V at reducing marine debris on Australian beaches.

To mitigate marine debris and promote sustainable marine industries, legislation and regulations surrounding the management of marine debris have been adopted worldwide. One of the most well-known and important agreements is the International Convention for the Prevention of Pollution from Ships (MARPOL), which focuses on reducing all types of ship-sourced marine pollution. MARPOL Annex V, which deals with the disposal of solid waste, came into force on 31 December 1988. However, was only amended to include a complete ban on waste disposal as of 1 January 2013. Assessing the effectiveness of key regulations is fundamental for supporting evidence-based decisions regarding the management of our oceans. Here, we evaluated whether MARPOL Annex V translated into a decrease in the incidence of shipping- and commercial fishing-sourced debris on remote beaches in Australia using 14 years of standardised, community-driven data. From 2006 to 2020 there was a significant change over time in the density of fishing and shipping debris on Australian beaches; debris density increased up to 2013 followed by a decrease until mid-2017. Although the new regulation started in January 2013, the decrease in density was not recorded until one year later. The decline was only observed for 4 years, reinforcing the existence of lags between the implementation of international agreements and the corresponding potential reduction in debris in the environment. This provides compelling evidence that international agreements and policies by themselves are not enough to solve the debris problem, with improved implementation and enforcement also required. We discuss future perspectives and solutions to reduce ocean-sourced litter inputs into the ocean and highlight the urgent need for action.

'Plasticosis': Characterising macro- and microplastic-associated fibrosis in seabird tissues.

As biota are increasingly exposed to plastic pollution, there is a need to closely examine the sub-lethal 'hidden' impacts of plastic ingestion. This emerging field of study has been limited to model species in controlled laboratory settings, with little data available for wild, free-living organisms. Highly impacted by plastic ingestion, Flesh-footed Shearwaters (Ardenna carneipes) are thus an apt species to examine these impacts in an environmentally relevant manner. A Masson's Trichrome stain was used to document any evidence of plastic-induced fibrosis, using collagen as a marker for scar tissue formation in the proventriculus (stomach) of 30 Flesh-footed Shearwater fledglings from Lord Howe Island, Australia. Plastic presence was highly associated with widespread scar tissue formation and extensive changes to, and even loss of, tissue structure within the mucosa and submucosa. Additionally, despite naturally occurring indigestible items, such as pumice, also being found in the gastrointestinal tract, this did not cause similar scarring. This highlights the unique pathological properties of plastics and raises concerns for other species impacted by plastic ingestion. Further, the extent and severity of fibrosis documented in this study gives support for a novel, plastic-induced fibrotic disease, which we define as 'Plasticosis,'.

Can the mass of plastic ingested by seabirds be predicted by the number of ingested items?

Plastics pollution has been documented for decades, yet repeatable methods for evaluating quantities are lacking. For wildlife, the mass and number of ingested plastics are widely reported, but these are not without their challenges, especially in field settings. Rapid methods for estimating the mass of ingested plastic could therefore be useful, but the relationship with the number of ingested pieces has not been explored. Using a dataset covering 1278 individuals of 11 Procellariiform species, we investigated this relationship to determine if counts could act as a proxy for the mass of ingested plastic by seabirds. Larger species ingested larger pieces of plastic, and birds that consumed more pieces also ingested items that are physically larger. Across species, sample size significantly influenced the slope of the relationship between the mass and number of ingested plastics. The mass-number relationship is species-specific, highly driven by sample size, and varies temporally.

Foraging strategy influences the quantity of ingested micro- and nanoplastics in shorebirds.

Coastlines, including estuaries, mudflats, and beaches, are particularly susceptible to plastic pollution, which can accumulate from both marine and terrestrial sources. While numerous studies have confirmed the presence of microplastics (1-5 mm) along coastlines, few have focused on very small particles (<1 µm) or quantified exposure within the organisms that inhabit these areas, such as shorebirds. Here, we quantified small plastics (200 nm-70 µm) in two resident shorebird species in Tasmania, and compared this to quantities found in the surrounding sediments in order to investigate the potential exposure and transfer of particles within these ecosystems. Analysis was performed using a combination of flow cytometry for quantification of micro- and nanoplastics (200 nm-70 µm), and µm-FT-IR for validation and polymer identification of particles >5.5 × 5.5 µm. Micro- and nano-plastics were detected in 100% of guano samples from surface-feeding Eastern Hooded Plovers (Thinornis cucullatus) and 90% of Australian Pied Oystercatcher (Haematopus longirostris) guano, a species that forages for coastal invertebrates at 60-90 mm depth, and 100% of beach sediments. Hooded Plover guano contained 32 × more plastics, on average, than Pied Oystercatcher guano. Interestingly, the abundance of plastic particles within sediments collected from shorebird foraging sites did not appear to have a significant effect on the number of plastics the birds had ingested, suggesting the difference between species is likely a result of other variables, such as prey selection. The results of this study highlight the importance of including techniques that provide quantitative data on the abundance and size of the smallest possible particle sizes, and demonstrate the significant proportion of small plastics that are 'missed' using standard analysis tools.

New Methods for the Quantification of Ingested Nano- and Ultrafine Plastics in Seabirds.

Plastic ingestion has been documented in a plethora of taxa. However, there is a significant gap in the detection of nano- and ultrafine particles due to size limitations of commonly used techniques. Using two Australian seabird species as case studies, the flesh-footed shearwater (FFSH) Ardenna carneipes and short-tailed shearwater (STSH) A. tenuirostris, we tested a novel approach of flow cytometry to quantify ingested particles <70 µm in the fecal precursor (guano; colon and cloacal contents) of both species. This method provided the first baseline data set for these species for plastics in the 200 nm-70 µm particle size ranges and detected a mean of 553.50 ± 91.21 and 350.70 ± 52.08 plastics (count/mg fecal precursor, wet mass) in STSH and FFSH, respectively, whereas Fourier transform infrared spectroscopy (FT-IR) provided accurate measurements of polymer compositions and quantities in the size range above 5.5 × 5.5 µm2. The abundance of nano- and ultrafine particles in the guano (count/mg) was not significantly different between species (p-value = 0.051), suggesting that foraging distribution or prey items, but not species, may contribute to the consumption of small plastics. In addition, there was no correlation between macroplastics in the stomach compared to the fecal precursor, indicating that small particles are likely bioaccumulating (e.g., through shedding and digestive fragmentation) and/or being directly ingested. Combining flow cytometry with FT-IR provides a powerful quantitative and qualitative analysis tool for detecting particles orders of magnitude smaller than that are currently explored with wider applications across taxa and marine environments.

The one-two punch of plastic exposure: Macro- and micro-plastics induce multi-organ damage in seabirds.

Plastic pollution in the world's oceans is ubiquitous and increasing. The environment is inundated with microplastics (< 1 mm), and the health effects of these less conspicuous pollutants is poorly known. In addition, there is now evidence that macroplastics can release microplastics in the form of shedding or digestive fragmentation, meaning there is potential for macroplastic exposure to induce direct and indirect pathology through microplastics. Therefore, there is an urgent need for data from wild populations on the relationship between macro- and microplastic exposure and the potential compounding pathological effects of these forms of plastics. We investigated the presence and impact of microplastics in multiple tissues from Flesh-footed Shearwaters Ardenna carneipes, a species that ingests considerable quantities of plastics, and used histopathological techniques to measure physiological responses and inflammation from the plastics. All organs examined (kidney, spleen, proventriculus) had embedded microplastic particles and this correlated with macroplastic exposure. Considerable tissue damage was recorded, including a significant reduction in tubular glands and rugae in the proventriculus, and evidence of inflammation, fibrosis, and loss of organ structures in the kidney and spleen. This indicates macroplastics can induce damage directly at the site of exposure, while microplastics can be mobilised throughout the body causing widespread pathology. Collectively, these results indicate the scope and severity of the health impacts of plastic pollution may be grossly underestimated.

Ingested plastics in beach-washed Fairy Prions Pachyptila turtur from Tasmania.

Plastic is an omnipresent pollutant in marine ecosystems and is widely documented to be ingested among seabird species. Procellariiformes are particularly vulnerable to plastic ingestion, which can cause internal damage, starvation, and occasionally mortality. In this study, 34 fledgling Fairy Prions (Pachyptila turtur) recovered during a wreck event in south-eastern Tasmania in 2022 were examined for ingested plastics and body condition (e.g., wing chord length). While many of the birds exhibited poor body condition, this was not correlated with the count or mass of ingested plastics. We hypothesise the marine heatwave event, and resulting lack of prey, contributed to bird body condition and subsequent mortality. We provide some of the first data on the size of individual plastic particles ingested by seabirds and make recommendations for future studies to report this important metric in a consistent manner that ensures data are comparable.

The influence of seabirds on their breeding, roosting and nesting grounds: A systematic review and meta-analysis.

Seabird species world-wide are integral to both marine and terrestrial environments, connecting the two systems by transporting vast quantities of marine-derived nutrients and pollutants to terrestrial breeding, roosting and nesting grounds via the deposition of guano and other allochthonous inputs (e.g. eggs, feathers). We conducted a systematic review and meta-analysis and provide insight into what types of nutrients and pollutants seabirds are transporting, the influence these subsidies are having on recipient environments, with a particular focus on soil, and what may happen if seabird populations decline. The addition of guano to colony soils increased nutrient levels compared to control soils for all seabirds studied, with cascading positive effects observed across a range of habitats. Deposited guano sometimes led to negative impacts, such as guanotrophication, or guano-induced eutrophication, which was often observed where there was an excess of guano or in areas with high seabird densities. While the literature describing nutrients transported by seabirds is extensive, literature regarding pollutant transfer is comparatively limited, with a focus on toxic and bioaccumulative metals. Research on persistent organic pollutants and plastics transported by seabirds is likely to increase in coming years. Studies were limited geographically, with hotspots of research activity in a few locations, but data were lacking from large regions around the world. Studies were also limited to seabird species listed as Least Concern on the IUCN Red List. As seabird populations are impacted by multiple threats and steep declines have been observed for many species world-wide, gaps in the literature are particularly concerning. The loss of seabirds will impact nutrient cycling at localized levels and potentially on a global scale as well, yet it is unknown what may truly happen to areas that rely on seabirds if these populations disappear.

Comparing methods for monitoring nest debris using silver gulls as a case study.

Global plastics production is increasing exponentially and contributing to significant pollution of the marine environment. Of particular concern is ingestion and entanglement risks for marine wildlife, including when items such as rope are incorporated into nest structures. These events are commonly documented using photographic and visual surveys, and each presents a number of challenges and benefits for species conservation and monitoring. Here we compare an invasive (i.e., removing debris from nests) and non-invasive (i.e., photographs) sampling method for quantifying nest debris using the silver gull (Chroicocephalus novahollandiae) as a case study. Overall, 17 debris items were detected in 9% of gull nests. While the use of photographs to monitor nest debris is increasingly popular, the invasive method detected one additional debris item not identified using photography. We therefore recommend caution for nest debris and other monitoring programs where identifying small or cryptic items may require a high level of skill.

Complete Mitochondrial Genome Sequence of a Seabird, Wedge-Tailed Shearwater (Ardenna pacifica).

Here, we report the complete mitochondrial genome sequence of a seabird, wedge-tailed shearwater (Ardenna pacifica). The circular genome has a size of 16,434 bp and contains 13 protein-coding genes, 22 tRNA genes, and 2 rRNA genes. The study provides a reference mitochondrial genome of wedge-tailed shearwater for further molecular studies.

Cleaner seas: reducing marine pollution.

In the age of the Anthropocene, the ocean has typically been viewed as a sink for pollution. Pollution is varied, ranging from human-made plastics and pharmaceutical compounds, to human-altered abiotic factors, such as sediment and nutrient runoff. As global population, wealth and resource consumption continue to grow, so too does the amount of potential pollution produced. This presents us with a grand challenge which requires interdisciplinary knowledge to solve. There is sufficient data on the human health, social, economic, and environmental risks of marine pollution, resulting in increased awareness and motivation to address this global challenge, however a significant lag exists when implementing strategies to address this issue. This review draws upon the expertise of 17 experts from the fields of social sciences, marine science, visual arts, and Traditional and First Nations Knowledge Holders to present two futures; the Business-As-Usual, based on current trends and observations of growing marine pollution, and a More Sustainable Future, which imagines what our ocean could look like if we implemented current knowledge and technologies. We identify priority actions that governments, industry and consumers can implement at pollution sources, vectors and sinks, over the next decade to reduce marine pollution and steer us towards the More Sustainable Future. Supplementary Information: The online version contains supplementary material available at 10.1007/s11160-021-09674-8.

Plastic and natural inorganic microparticles do not differ in their effects on adult mussels (Mytilidae) from different geographic regions.

Microplastics are ubiquitous in the marine environment and studies on their effects on benthic filter feeders at least partly revealed a negative influence. However, it is still unclear whether the effects of microplastics differ from those of natural suspended microparticles, which constitute a common stressor in many coastal environments. We present a series of experiments that compared the effects of six-week exposures of marine mussels to two types of natural particles (red clay and diatom shells) to two types of plastic particles (Polymethyl Methacrylate and Polyvinyl Chloride). Mussels of the family Mytilidae from temperate regions (Japan, Chile, Tasmania) through subtropical (Israel) to tropical environments (Cabo Verde) were exposed to concentrations of 1.5 mg/L, 15 mg/L and 150 mg/L of the respective microparticles. At the end of this period, we found significant effects of suspended particles on respiration rate, byssus production and condition index of the animals. There was no significant effect on clearance rate and survival. Surprisingly, we observed only small differences between the effects of the different types of particles, which suggests that the mussels were generally equally robust towards exposure to variable concentrations of suspended solids regardless of whether they were natural or plastic. We conclude, that microplastics and suspended solids elicit similar effects on the tested response variables, and that both types of microparticles mainly cause acute responses rather than more persistent carry-over effects.

A baseline study of per- and polyfluoroalkyl substances (PFASs) in waterfowl from a remote Australian environment.

Elevated concentrations of PFASs in the liver may pose a toxicological risk to bird species and humans that consume them. This study aimed to determine concentrations of 43 per- and polyfluoroalkyl substances (PFASs) in livers (n = 80) of Australian Shelducks (Tadorna tadornoides), Pacific Black Ducks (Anas superciliosa), and Teals (Anas sp.), as well as water and sediment from a remote Australian environment. Maximum concentrations of PFBA (1.9 ng L-1), PFOA (1.7 ng L-1) and PFOS (0.99 ng L-1) in water were consistent with long-range atmospheric and oceanic transport. PFOS (30%) and PFNA (22%) were the most frequently detected PFASs in Australian Shelduck livers (0.31 ± 0.68 ng g-1 ww and 0.16 ± 0.15 ng g-1 ww respectively). Maximum concentrations of PFOS in Pacific Black Ducks (50%) and Teals (44%) was 2.4 ng g-1 ww and 5.3 ng g-1 ww respectively. While PFAS levels in birds from this remote environment were below current animal consumption guidelines, continued monitoring of this ecosystem is recommended to assess the human health risk of consumption of wild game.

Temporal trends and interannual variation in plastic ingestion by Flesh-footed Shearwaters (Ardenna carneipes) using different sampling strategies.

The world's oceans are under increasing pressure from anthropogenic activities, including significant and rapidly increasing inputs of plastic pollution. Seabirds have long been considered sentinels of ocean health, providing data on physical and chemical pollutants in their marine habitats. However, long-term data that can elucidate important patterns and changes in seabird exposure to marine pollutants are relatively limited but are urgently needed to identify and support effective policy measures to reduce plastic waste. Using up to 12 years of data, we examined the benefits and challenges of different approaches to monitoring plastic in seabirds, and the relationship between plastic and body size parameters. We found the mass and number of ingested plastics per bird varied by sample type, with lavage and road-kill birds containing less plastic (9.17-9.33 pieces/bird) than beach-washed or otherwise dead birds (27.62-32.22 pieces/bird). Beached birds therefore provide data for only a particular subset of the population, mostly individuals in poorer body condition, including those severely impacted by plastics. In addition, the mass and number of plastics in beached birds were more variable, therefore the sample sizes required to detect a change in plastic over time were significantly larger than for lavaged birds. The use of lavaged birds is rare in studies of plastic ingestion due to ethical and methodological implications, and we recommend future work on ingested plastics should focus on sampling this group to ensure data are more representative of a population's overall exposure to plastics.

Plastic debris increases circadian temperature extremes in beach sediments.

Plastic pollution is the focus of substantial scientific and public interest, leading many to believe the issue is well documented and managed, with effective mitigation in place. However, many aspects are poorly understood, including fundamental questions relating to the scope and severity of impacts (e.g., demographic consequences at the population level). Plastics accumulate in significant quantities on beaches globally, yet the consequences for these terrestrial environments are largely unknown. Using real world, in situ measurements of circadian thermal fluctuations of beach sediment on Henderson Island and Cocos (Keeling) Islands, we demonstrate that plastics increase circadian temperature extremes. Particular plastic levels were associated with increases in daily maximum temperatures of 2.45°C and decreases of daily minimum by - 1.50°C at 5 cm depth below the accumulated plastic. Mass of surface plastic was high on both islands (Henderson: 571 ± 197 g/m2; Cocos: 3164 ± 1989 g/m2), but did not affect thermal conductivity, specific heat capacity, thermal diffusivity, or moisture content of beach sediments. Therefore, we suggest plastic effects sediment temperatures by altering thermal inputs and outputs (e.g., infrared radiation absorption). The resulting circadian temperature fluctuations have potentially significant implications for terrestrial ectotherms, many of which have narrow thermal tolerance limits and are functionally important in beach habitats.

A standardised method for estimating the level of visible debris in bird nests.

Unlike records of plastic ingestion and entanglement in seabirds which date back to the 1960s, the literature regarding debris in bird nests is comparatively limited. It is important to identify standardised methods early so that data are collected in a consistent manner, ensuring that future studies can be comparable. Here, we outline a method that can be applied to photographs for estimating the proportion of visible debris at the surface of a nest. This method uses ImageJ software to superimpose a grid onto a photograph of a nest/s. The number of cells with and without debris are then counted. Our proposed method is repeatable, straightforward, and accessible. We optimised the method to estimate the level of visible debris in Northern Gannet (Morus bassanus) nests, however, with some modification (i.e., adjustment of grid cell size), it could be applied to other seabird species, and terrestrial birds, that incorporate debris within nests.

Measuring nest incorporation of anthropogenic debris by seabirds: An opportunistic approach increases geographic scope and reduces costs.

Data on the prevalence of anthropogenic debris in seabird nests can be collected alongside other research or through community science initiatives to increase the temporal and spatial scale of data collection. To assess the usefulness of this approach, we collated data on nest incorporation of debris for 14 seabird species from 84 colonies across five countries in northwest Europe. Of 10,274 nests monitored 12% contained debris, however, there was large variation in the proportion of nests containing debris among species and colonies. For several species, the prevalence of debris in nests was significantly related to the mean Human Footprint Index (HFI), a proxy for human impact on the environment, within 100 km of the colony. Collecting opportunistic data on nest incorporation of debris by seabirds provides a cost-effective method of detecting changes in the prevalence of debris in the marine environment across a large geographic scale.