Biodegradable microplastics: Uptake by and effects on the rockpool shrimp Palaemon elegans (Crustacea: Decapoda).

Ingestion of microplastics can lead to deleterious consequences for organisms, as documented by numerous laboratory studies. The current knowledge is based on a multitude of effect studies, conducted with conventional fossil-based and non-degradable plastics. However, there is a lack of information about the acceptance and the effects of novel bio-based and biodegradable plastics. Biodegradable plastics are considered an alternative to conventional plastics and are showing rapidly growing production rates. Biodegradable plastics can disperse into the environment in the same way as conventional plastics do, becoming available to marine organisms. This study aims to provide new insights into the uptake and effects of biodegradable microplastics on marine invertebrates. Rockpool shrimp, Palaemon elegans, were fed with algal flakes coated with polylactic acid (PLA), polyhydroxybutyrate-co-valerate (PHBV) and conventional low-density polyethylene (LDPE) microparticles. Live observations showed that all of the different types of microplastics were ingested. After dissection of the shrimp, less LDPE particles were found in the stomachs than PLA and PHBV particles. This indicates a longer retention time of biodegradable microplastics compared to conventional microplastics. Presumably, less LDPE particles were ingested or evacuated from the stomach, probably by regurgitation. The ingestion of microparticles of all types of plastics induced enzymatic activity of short-chain carboxylesterases in the midgut glands of the shrimp. However, only PLA induced enzymatic activity of medium-chain carboxylesterases. Palaemon elegans showed no oxidative stress response after ingestion of microparticles, irrespective of polymer type. From our results we conclude that biodegradable plastics might have different effects than conventional plastics. The longer retention times of biodegradable plastics might enhance exposure to leaching additives and other harmful substances. Our study provides new insights into how biodegradable plastics might affect aquatic fauna and indicate that the use of biodegradable plastics needs to be reconsidered to some extent.

Impacts of exhaust gas cleaning systems (EGCS) discharge waters on planktonic biological indicators.

Exhaust Gas Cleaning Systems (EGCS), operating in open-loop mode, continuously release acidic effluents (scrubber waters) to marine waters. Furthermore, scrubber waters contain high concentrations of metals, polycyclic aromatic hydrocarbons (PAHs), and alkylated PAHs, potentially affecting the plankton in the receiving waters. Toxicity tests evidenced significant impairments in planktonic indicators after acute, early-life stage, and long-term exposures to scrubber water produced by a vessel operating with high sulphur fuel. Acute effects on bacterial bioluminescence (Aliivibrio fischeri), algal growth (Phaeodactylum tricornutum, Dunaliella tertiolecta), and copepod survival (Acartia tonsa) were evident at 10 % and 20 % scrubber water, while larval development in mussels (Mytilus galloprovincialis) showed a 50 % reduction at ∼5 % scrubber water. Conversely, larval development and reproductive success of A. tonsa were severely affected at scrubber water concentrations ≤1.1 %, indicating the risk of severe impacts on copepod populations which in turn may result in impairment of the whole food web.

Comparison of pre-treatment methods and heavy density liquids to optimize microplastic extraction from natural marine sediments.

The ubiquitous occurrence of anthropogenic particles, including microplastics in the marine environment, has, over the last years, gained worldwide attention. As a result, many methods have been developed to estimate the amount and type of microplastics in the marine environment. However, there are still no standardized protocols for how different marine matrices should be sampled or how to extract and identify these particles, making meaningful data comparison hard. Buoyant microplastics are influenced by winds and currents, and concentrations could hence be expected to be highly variable over time. However, since both high density and most of the initially buoyant microplastics are known to eventually sink and settle on the seafloor, marine sediments are proposed as a suitable matrix for microplastics monitoring. Several principles, apparatuses, and protocols for extracting microplastics from marine sediments have been presented, but extensive comparison of the different steps in the protocols using real environmental samples is lacking. Thus, in this study, different pre-treatment and subsequent density separation protocols for extraction of microplastics from replicate samples of marine sediment were compared. Two pre-treatment methods, one using inorganic chemicals (NaClO + KOH + Na4P2O7) and one using porcine pancreatic enzymes, as well as one with no pre-treatment of the sediment, were compared in combination with two commonly used high-density saline solutions used for density separation, sodium chloride (NaCl) and zinc chloride (ZnCl2). Both pre-treatment methods effectively removed organic matter, and both saline solutions extracted lighter plastic particles such as polyethylene (PE) and polypropylene (PP). The most efficient combination, chemical pre-treatment and density separation with ZnCl2, was found to extract > 15 times more particles (≥ 100 µm) from the sediment than other treatment combinations, which could largely be explained by the high presence and efficient extraction of PVC particles.

Severe Toxic Effects on Pelagic Copepods from Maritime Exhaust Gas Scrubber Effluents.

To reduce sulfur emission from global shipping, exhaust gas cleaning systems are increasingly being installed on board commercial ships. These so-called scrubbers extract SOX by spraying water into the exhaust gas. An effluent is created which is either released directly to the sea (open-loop system) or treated to remove harmful substances before release (closed-loop system). We found severe toxic effects in the ubiquitous planktonic copepod Calanus helgolandicus of exposure to effluents from two closed-loop systems and one open-loop system on North Sea ships. The effluents contained high concentrations of heavy metals and polycyclic aromatic hydrocarbons (PAHs), including alkylated PAHs. We observed significantly elevated mortality rates and impaired molting already in the lowest tested concentrations of each effluent: 0.04 and 0.1% closed-loop effluents and 1% open-loop effluent. These concentrations correspond to total hydrocarbon concentrations of 2.8, 2.0, and 3.8 µg L-1, respectively, and compared to previous studies on oil toxicity in copepods, scrubber effluents appear more toxic than, for example, crude oil. None of the individual PAHs or heavy metals analyzed in the effluents occurred in concentrations which could explain the high toxicity. The effluents showed unexpected alkylated PAH profiles, and we hypothesize that scrubbers act as witch's cauldrons where undesired toxic compounds form so that the high toxicity stems from compounds we know very little about.

A temporal record of microplastic pollution in Mediterranean seagrass soils.

Plastic pollution is emerging as a potential threat to the marine environment. In the current study, we selected seagrass meadows, known to efficiently trap organic and inorganic particles, to investigate the concentrations and dynamics of microplastics in their soil. We assessed microplastic contamination and accumulation in 210Pb dated soil cores collected in Posidonia oceanica meadows at three locations along the Spanish Mediterranean coast, with two sites located in the Almería region (Agua Amarga and Roquetas) and one at Cabrera Island (Santa Maria). Almería is known for its intense agricultural industry with 30 000 ha of plastic-covered greenhouses, while the Cabrera Island is situated far from urban areas. Microplastics were extracted using enzymatic digestion and density separation. The particles were characterized by visual identification and with Fourier-transformed infrared (FTIR) spectroscopy, and related to soil age-depth chronologies. Our findings showed that the microplastic contamination and accumulation was negligible until the mid-1970s, after which plastic particles increased dramatically, with the highest concentrations of microplastic particles (MPP) found in the recent (since 2012) surface soil of Agua Amarga (3819 MPP kg-1), followed by the top-most layers of the soil of the meadows in Roquetas (2173 kg-1) and Santa Maria (68-362 kg-1). The highest accumulation rate was seen in the Roquetas site (8832 MPP m-2 yr-1). The increase in microplastics in the seagrass soil was associated to land-use change following the intensification of the agricultural industry in the area, with a clear relationship between the development of the greenhouse industry in Almería and the concentration of microplastics in the historical soil record. This study shows a direct linkage between intense anthropogenic activity, an extensive use of plastics and high plastic contamination in coastal marine ecosystems such as seagrass meadows. We highlight the need of proper waste management to protect the coastal environment from continuous pollution.

Summer sea ice melt and wastewater are important local sources of microlitter to Svalbard waters.

Human activities leave traces of marine litter around the globe. The Arctic is, despite its remoteness, emerging as an area of no exception to this environmental issue. Arctic sea ice has previously been found to constitute a temporal sink of microplastics, but the potential release and subsequent fate of microplastics in the marine environment are yet unknown. Furthermore, the relative importance of local sources of microplastics in the Arctic marine environment is under discussion. In this study, the concentration and distribution of anthropogenic microparticles (AMPs, <5 mm, including microplastics) have been investigated in marine waters and sea ice of Svalbard. Seawater samples throughout the water column and floating sea ice samples were collected along a transect originating in Rijpfjorden, reaching northwards to the sea ice-edge. Seawater samples were also collected along a transect extending westwards from head to mouth of Kongsfjorden. Samples were collected throughout the water column with stations positioned to enable detection of potential AMP emissions from the wastewater outlet in Ny-Ålesund. Along both transects, environmental parameters were measured to explore potential correlations with AMP distribution. High concentrations of AMPs were detected in sea ice (158 ±â€¯155 AMPs L-1). Based on both AMP concentrations and characteristics, AMPs identified in seawater of the marginal ice zone are to a large extent likely released during the melting of sea ice. The release of AMPs during summer melting of sea ice was concomitantly taking place with the ice-edge bloom, suggesting increased bioavailability to Arctic marine biota. Concentrations of AMPs were up to an order of magnitude higher in Kongsfjorden (up to 48.0 AMPs L-1) than in Rijpfjorden (up to 7.4 AMPs L-1). The distribution and composition of AMPs in Kongsfjorden suggest the wastewater outlet in Ny-Ålesund to be a likely source. Our results emphasize the importance of local point- and diffuse sources of AMPs in the Arctic and stress the urgency of considering their associated environmental impact. Implementation of regulatory policy is of importance, particularly since human activities and environmental pressures are increasing in the Arctic.

An efficient and gentle enzymatic digestion protocol for the extraction of microplastics from bivalve tissue.

Standardized methods for the digestion of biota for microplastic analysis are currently lacking. Chemical methods can be effective, but can also cause damage to some polymers. Enzymatic methods are known to be gentler, but often laborious, expensive and time consuming. A novel tissue digestion method with pancreatic enzymes and a pH buffer (Tris) is here presented in a comparison to a commonly applied digestion protocol with potassium hydroxide. The novel protocol demonstrates a highly efficient removal of bivalve tissue (97.7 ±â€¯0.2% dry weight loss) already over-night. Furthermore, it induces no impairment in terms of ability to correctly identify four pre-weathered plastic polymers and six textile fiber polymers by Fourier transform infrared spectroscopy after exposure. The high-throughput protocol requires minimal handling, is of low cost and does not pose risk to the performer or the environment. It is therefore suggested as a candidate for a standardized digestion protocol, enabling successful analysis of microplastics ingested by bivalves.

Microplastic in wild populations of the omnivorous crab Carcinus aestuarii: A review and a regional-scale test of extraction methods, including microfibres.

Microplastic (MP) has become ubiquitous in the marine environment. Its threat to marine organisms has been demonstrated under laboratory conditions, yet studies on wild populations still face methodological difficulties. We reviewed the methods used to separate MP from soft animal tissues and highlighted a lack of standardised methodologies, particularly critical for synthetic microfibres. We further compared enzymatic and a potassium hydroxide (KOH)-based alkaline digestion protocols on wild crabs (Carcinus aestuarii) collected from three coastal lagoons in the north Adriatic Sea and on laboratory-prepared synthetic polyester (PES) of different colour and polypropylene (PP). We compared the cost-effectiveness of the two methods, together with the potential for adverse quantitative or qualitative effects on MP that could alter the capability of the polymers to be recognised via microscopic or spectroscopic techniques. Only 5.5% of the 180 examined crabs contained MP in their gastrointestinal tracts, with a notably high quantitative variability between individuals (from 1 to 117 particles per individual). All MP found was exclusively microfibres, mainly PES, with a mean length (±SE) of 0.5 ±â€¯0.03 mm. The two digestion methods provided comparable estimates on wild crabs and did not cause any visible physical or chemical alterations on laboratory-prepared microfibres treated for up to 4 days. KOH solution was faster and cheaper compared to the enzymatic extraction, involving fewer procedural steps and therefore reducing the risk of airborne contamination. With digestion times longer than 4 days, KOH caused morphological alterations of some of the PES microfibres, which did not occur with the enzymatic digestion. This suggests that KOH is effective for the digestion of small marine invertebrates or biological samples for which shorter digestion time is required, while enzymatic extraction should be considered as alternative for larger organisms or sample sizes requiring longer digestion times.

Bioturbation-driven release of organic contaminants from Baltic Sea sediments mediated by the invading polychaete Marenzelleria neglecta.

Baltic Sea sediments are among the world's most polluted regarding eutrophication and contamination. Eutrophication-induced hypoxia has caused depletion of bioturbating macrofauna in vast areas, producing laminated sediments. We investigated if reoxygenation and colonization by the invading deep-burrowing polychaete Marenzelleria neglecta may cause an augmented contaminant release from Baltic Sea sediments. Intact laminated sediment cores were exposed either to in situ hypoxia, reoxygenation, or reoxygenation combined with bioturbating M. neglecta. The release fluxes of particle-associated (N(Pat)) and dissolved (N(Diss)) PCBs and chlorinated pesticide residues (POPs) were quantified (GC-ECD) after 85 d along with contaminant concentrations in sediment and biota. Lavoisier-based mass transfer coefficients (Kf) were calculated from N(Diss). Sediment contaminant concentrations were high (sigmaPCB7: 42-52 ng g(sediment)(-1) dw) due to emissions from Stockholm. N(Diss) always exceeded N(Part) by an order of magnitude. Bioturbation enhanced N(Diss) and Kf from hypoxic sediments 0.7-3 times while reoxygenation alone had no significant effect. M. neglecta accumulated low amounts of contaminants but significantly stimulated aquatic release of bioavailable sequestered contaminants. Bioturbation should be included in aquatic contaminant fate models. We advise to consider quiescent pollutant sources and possible ecological shifts when aiming to restore eutrophicated aquatic environments.

Effects of sediment organic matter quality on bioaccumulation, degradation, and distribution of pyrene in two macrofaunal species and their surrounding sediment.

Sediment dwelling macrofauna (infauna) are important vectors for the transfer of sediment-associated contaminants to higher trophic levels. Sedimenting organic matter constitutes an important food source for all benthic organisms and changes seasonally in terms of quantity and quality. Sediment organic matter (SOM) quality affects organism activity and feeding behaviour, and is therefore also likely to affect contaminant fate in benthic systems. We investigated the impact of SOM quality (enrichment with either labile Tetraselmis sp. or refractory lignin) on the accumulation and metabolism of sediment-associated pyrene in Nereis diversicolor (Annelida) and Amphiura filiformis (Echinodermata), as well as the combined effect of SOM quality and infaunal bioturbation on pyrene distribution and metabolism in the sediment. After 45 d of exposure, SOM quality almost doubled pyrene bioaccumulation in both species, while pyrene metabolism remained unaffected. Metabolites comprised approximately 80% of the total tissue pyrene in N. diversicolor and approximately 40% in A. filiformis. A. filiformis arms contained one fifth of the disk pyrene concentration. Approximately 20% of the pyrene found in A. filiformis arms was found to be covalently bound to, e.g. DNA, RNA or proteins, thus reducing pyrene bioavailability to arm-cropping predators. The sedimentary pyrene distribution and metabolism was species-dependent, but correlated poorly with prevailing knowledge on species-specific bioturbation patterns. This was attributed to the comparably high biodegradability of the contaminant thus altering its sorptive characteristics and function as inert tracer. Subduction of pyrene and metabolites occurred, and the fraction of pyrene covalently bound to SOM increased with depth, thereby removing pyrene from the bioavailable pool. Our results imply that bioaccumulation and trophic transfer of sediment-associated PAH should increase following fresh organic matter input, e.g. after sedimentation of phytoplankton blooms. We stress the importance of considering behavioural characteristics of infauna and the trophic situation of the system when assessing fate and effects of sediment-associated contaminants.

Role of sediment organic matter quality and feeding history in dietary absorption and accumulation of pyrene in the mud snail (Hydrobia ulvae).

Organic matter (OM) input to marine sediments varies seasonally both in quantity and quality. Because sedimentary OM (SOM) constitutes food for many benthic organisms, its properties should affect the dietary uptake of sediment-associated contaminants. We explored the effect of SOM quality/food value on short- and long-term pyrene accumulation in the mud snail (Hydrobia ulvae) and performed dual-tracer pulse-chase experiments to investigate the feeding mechanisms driving dietary pyrene uptake. The quality of the SOM was manipulated by enriching sediments either with high-quality microalgae or low-quality lignin, adding equal amounts of total organic carbon. Long- and short-term bioaccumulation increased with increasing SOM quality, as did pyrene ingestion rate (IR(pyr)), which also was affected by feeding history. By feeding selectively, snails concentrated pyrene 10-fold in ingested compared to ambient sediment, independent of SOM quality. Average pyrene absorption efficiency (AE(pyr): -65%) varied inversely with SOM quality and IR(pyr). Both AE(pyr) and gut passage time (alpha 1/IR(pyr)) agreed with theoretical models incorporating the time-dependence of absorption efficiency. Thus, SOM quality moderates dietary contaminant uptake in deposit feeders, and in H. ulvae, this occurs via OM-induced alterations of ingestion rate. Consequently, enhanced sediment-associated contaminant uptake is predicted for deposit feeders following phytoplankton blooms, principally because of OM quality-driven increases in the ingestion rate.

Comparison of different IMAC techniques used for enrichment of phosphorylated peptides.

Four commercially available immobilized metal ion affinity chromatography (IMAC) methods for phosphopeptide enrichment were compared using small volumes and concentrations of phosphopeptide mixtures with or without extra-added bovine serum albumin (BSA) nonphosphorylated peptides. Addition of abundant tryptic BSA peptides to the phosphopeptide mixture increases the demand for selective IMAC capture. While SwellGel gallium Discs, IPAC Metal Chelating Resin, and ZipTipMC Pipette Tips allow for the possibility of enriching phosphopeptides, the Gyrolab MALDI IMAC1 also presents the possibility of verifying existing phosphopeptides after a dephosphorylation step. Phosphate-containing peptides are identified through a mass shift between phosphorylated and dephosphorylated spectra of 80 Da (or multiples of 80 Da). This verification is useful if the degree of phosphorylation is low in the sample or if the ionization is unfavorable, which often is the case for phosphopeptides. A peptide mixture in which phosphorylated serine, threonine, and tyrosine were represented was diluted in steps and thereafter enriched using the four different IMAC methods prior to analyses with matrix assisted laser desorption/ionization mass spectrometry. The enrichment of phosphopeptides using SwellGel Gallium Discs or Gyrolab MALDI IMAC1 was not significantly affected by the addition of abundant BSA peptides added to the sample mixture, and the achieved detection limits using these techniques were also the lowest. All four of the included phosphopeptides were detected by MALDI-MS only after enrichment using the Gyrolab MALDI IMAC1 compact disc (CD) and detection down to low femtomole levels was possible. Furthermore, selectivity, reproducibility, and detection for a number of other phosphopeptides using the IMAC CD are reported herein. For example, two phosphopeptides sent out in a worldwide survey performed by the Proteomics Research Group (PRG03) of the Association of Biomolecular Resource Facilities (ABRF) were detected and verified by means of the 80 Da mass shift achieved by on-column dephosphorylation.

Impact of sediment organic matter quality on the fate and effects of fluoranthene in the infaunal brittle star Amphiura filiformis.

Hydrophobic contaminants, such as polycyclic aromatic hydrocarbons (PAHs) readily adsorb to organic matter. The aim of this study was to determine the importance of the quality of sedimentary organic matter for the uptake, biotransformation and toxicity of the PAH, fluoranthene (Flu), in the infaunal brittle star Amphiura filiformis. Brittle stars were exposed to a base sediment covered by a 2 cm Flu-spiked top layer (30 microg Flu/g dry wt. sed.), enriched to the same total organic carbon content with either refractory or labile organic matter. The labile carbon source was concentrated green flagellate: Tetraselmis spp. The refractory carbon source was lignin from a paper mill. Tissue concentrations of Flu both in disk and arm-fractions were determined as total Flu, parent Flu (i.e. untransformed), aqueous Flu-metabolites, polar Flu-metabolites and tissue residue Flu (i.e. unextractable). Our results showed that sediment particle ingestion is a pathway by which Flu can enter benthic food webs. Flu toxicity (measured as arm-regeneration), but not net accumulation, was dependent on the nutritional quality of the ingested sediment particles. Flu bioaccumulation could not be attributed solely to equilibrium partitioning between organism lipid content and organic content of the sediment. Biotransformation of Flu by brittle stars was very limited and unaffected by organic matter quality. A. filiformis contributed to the downward transport of Flu from the surface sediment to the burrow lining. The limited breakdown of parent Flu by brittle stars and/or microorganisms was relatively higher in burrows compared to surface sediment, and highest in the presence of labile organic matter. Tissue concentrations were higher in disk than in arms, but the proportion of metabolic products relative to parent Flu was higher in arms than in the disk fraction. We estimate that the yearly mobilization of sediment-associated Flu by arm-regeneration in A. filiformis is in the range of 3.8-29.4 microg total Flu eq. m(-2) year(-1) at a sediment concentration of 30 microg Flu/g dry wt. sed.