In the marine environment, microplastic contamination and acidification may occur simultaneously, this study evaluated the effects of ocean acidification and microplastics on oxidative stress responses and digestive enzymes in mussels. The thick shell mussels Mytilus coruscus were exposed to four concentrations of polystyrene microspheres (diameter 2 μm, 0, 10, 10⁴ and 10⁶ particles/L) under two pH levels (7.7 and 8.1) for 14 days followed by a 7-day recovery acclimation. Throughout the experiment, we found that microplastics and ocean acidification exerted little oxidative stress to the digestive gland. Only catalase (CAT) and glutathione (GSH) showed a significant increase along with increased microplastics during the experiment, but recovered to the control levels once these stressors were removed. No significant effects of pH and microplastics on glutathione peroxidase (GPx) and superoxide dismutase (SOD) were observed. The responses of digestive enzymes to both stressors were more pronounced than antioxidant enzymes. During the experiment, pepsin (PES), trypsin (TRS), alpha-amylase (AMS) and lipase (LPS) were significantly inhibited under microplastics exposure and this inhibition was aggravated by acidification conditions. Only PES and AMS tended to recover during the recovery period. Lysozyme (LZM) increased significantly under microplastic exposure conditions, but acidification did not exacerbate this effect. Therefore, combined stress of microplastics and ocean acidification slightly impacts oxidative responses but significantly inhibits digestive enzymes in mussels.

Microplastics are known to be associated with co-contaminants, but little is understood about the mechanisms by which these chemicals are transferred from ingested plastic to organisms. This study simulates marine avian gastric conditions in vitro to examine the bioaccessibility of authigenic metals (Fe, Mn) and trace metals (Co, Pb) that have been acquired by polyethylene microplastic pellets from their environment. Specifically, different categories of pellet were collected from beaches in Cornwall, southwest England, and exposed to an acidified saline solution of pepsin (pH ∼ 2.5) at 40 °C over a period of 168 h with extracted metal and residual metal (available to dilute aqua regia) analysed by ICP-MS. For Fe, Mn and Co, kinetic profiles consisted of a relatively rapid initial period of mobilisation followed by a more gradual approach to quasi-equilibrium, with data defined by a diffusion model and median rate constants ranging from about 0.0002 (μg L⁻¹)⁻¹ h⁻¹ for Fe to about 7 (μg L⁻¹)⁻¹ h⁻¹ for Co. Mobilisation of Pb was more complex, with evidence of secondary maxima and re-adsorption of the metal to the progressively modified pellet surface. At the end of the time-courses, maximum total concentrations were 38.9, 0.81, 0.014 and 0.10 μg g⁻¹ for Fe, Mn, Co and Pb, respectively, with maximum respective percentage bioaccessibilities of around 60, 80, 50 and 80. When compared with toxicity reference values for seabirds, the significance of metals acquired by microplastics from the environment and exposed to avian digestive conditions is deemed to be low, but studies of a wider range of plastics and metal associations (e.g. as additives) are required for a more comprehensive risk assessment.

Marine polychaetes and fish are known to ingest polystyrene microparticles in the environment. Laboratory microplastic feeding experiments have demonstrated that plastic may release endocrine-disrupting compounds such as diethylhexyl phthalate (DEHP), which can cause adverse effects in both vertebrates and invertebrates. In order to determine the influence of size and digestive conditions on the desorption of DEHP and other plasticizers to polychaetes and fish, we exposed polystyrene particles of various sizes under invertebrate and vertebrate digestive conditions (vertebrate mimic; pepsin, pH = 2.0, 24 °C, invertebrate mimic; Na taurocholate pH = 7, 18 °C). Estrogen receptor activation and concentrations of 12 plasticizers were measured in the extracts. DEHP, bisphenol S and 4-tert-octylphenol were the only compounds detected. Simulated vertebrate gut digestion did not significantly enhance the release of chemicals nor estrogenic activity. However, a 6.3 ± 2.0-fold increase in the concentration of DEHP was observed in extracts from invertebrate gut conditions (Mean ± SD; N = 24, p < 0.0001). Additionally, estimated particle surface area was positively correlated with estrogenic activity across all treatment types (r = 0.85, p < 0.0001). Overall, these data indicate an elevated bioaccessibility of DEHP may occur in invertebrates, and size-dependent desorption of uncharacterized estrogenic compounds from plastic suggest additional complexity when considering the risks of MP to aquatic organisms.

We describe a batch-extraction with simulated digestive fluid (salivary fluid, gastric fluid and intestinal fluid) to estimate the bioaccessibility of inhaled trace metals (TMs) in particulate matter less than 10 and 2.5 μm in aerodynamic diameter (PM₁₀ and PM₂.₅). Concentrations of the assayed TMs (As, Cd, Cr, Ni, Mn, Cu, Zn, Sb, Hg and Pb) were determined in PM₁₀ and PM₂.₅ samples by inductively coupled plasma-mass spectrometry. The TMs with the largest soluble fractions for airborne PM collected from winter and summer in saliva were Mn and Sb, respectively; in seasons this became Co in gastric fluid and Cu in intestinal fluid. Clearly, bioaccessibility is strongly dependent on particle size, the component of simulated digestive fluids (e.g., pH, digestive enzymes pepsin and trypsin), and the chemical properties of metal ions. The particle size and seasonal variation affected the inhaled bioaccessible fraction of PM-bound TMs during mucociliary clearance, which transported PM from the tracheal and the bronchial region to the digestive system. This study provides direct evidence for TMs in airborne PM being bioaccessible TMs are likely to possess an enhanced digestive toxic potential due to airborne PM pollution.

This work investigated the binding behavior of phenanthrene by humic acids (HA-2 and HA-5), proteins (bovine serum albumin (BSA)), lysozyme and pepsin), and their complexes using a passive dosing technique. All sorption isotherms were fitted well with Freundlich model and the binding capability followed an order of HA-5 > HA-2 > BSA > pepsin > lysozyme. In NaCl solution, phenanthrene binding to HA-BSA complexes was much higher than the sum of binding to individual HA and BSA, while there was no enhancement for HA-pepsin. Positively charged lysozyme slightly lowered phenanthrene binding on both HAs due to strong aggregation of HA-lysozyme complexes, leading to reduction in the number of binding sites. The binding enhancement by HA-BSA was observed under all tested ion species and ionic strengths. This enhancement can be explained by unfolding of protein, reduction of aggregate size and formation of HA-BSA complexes with favorable conformations for binding phenanthrene.

During two seasonal trawl surveys (April and October, 2012), red mullet specimens were caught from two sites of the northern Sicilian coast (Western Mediterranean), characterized by different degrees of pollution, to assess whether their digestive enzymes could be cost-effective diagnostic tools for endocrine disruption. Pepsin, chymotrypsin, carboxypeptidases A and B, amylase and lipase were measured in the digestive tract of each fish. During both samplings, significant differences in the digestive enzymatic patterns of fish collected from the two sites were found. In April, pepsin and lipase contents were significantly lower in fish from the most impacted site than in those from the reference site. In October, the enzymatic patterns showed trends different from spring, with controversial results for carboxypeptidases A and B and amylase. Pepsin and lipase patterns suggest a detrimental effect played by organic pollutants and the use of these enzymes as possible biomarkers of exposure to endocrine disruptors.

A simple methodology for the determination of bioavailability of fourteen metals in coastal sediments has been developed by simulating the conditions of digestive process of marine fishes. With this aim, a representative sediment composite sample was treated with hydrochloric acid solutions at different pH values, temperatures and contact times, in the presence and absence of Pepsin and Trypsin. The addition of Pepsin and Trypsin did not affect the extraction of most elements. As a result of the present study, the digestion with a hydrochloric acid solution at pH 1, 40°C and 12h is proposed. Adjustments of the temperature and time reaction could be made according to the specific ecosystem under study. The amount of metal extracted by other methods based on acetic acid was lower than that extracted by HCl treatment proposed.

An in vitro technique using simulated gastrointestinal (GI) fluids was applied to investigate the desorption of selected endocrine disrupting chemicals (EDCs), i.e. bisphenol A (BPA) and 17 α-ethinylestradiol (EE2), from the marine sediment in the digestive environment. The results show that the GI fluids suppressed chemical adsorption and greatly increased the desorption of BPA and EE2 from the sediment. Pepsin in the gastric fluid would compete for the adsorption sites with the adsorbates, and bile salts in the intestinal fluid had a solubilization effect on the chemicals. The amount of chemical release from the sediment in different fluids followed intestinal (fed)>intestinal (fasted)>gastric>saline water. During the dynamic desorption tests, 62% and 21% of sediment-bound BPA and EE2, respectively, could be released into the simulated GI fluids. The enhanced desorption of EDCs from sediment in the digestive system would make the pollutants more bioavailable in the ecosystem.

Adsorption/desorption and desorption hysteresis of methamphetamine (MMA) on carbon nanotubes (CNTs) as well as bioavailability of MMA were studied in simulated gastrointestinal fluids and background fluids. Adsorption of MMA in near-neutral (weak alkaline) intestinal fluid was enhanced, while adsorption of MMA on CNTs in acid gastric fluid was suppressed. Desorption of MMA is divided into fast and slow stages, and fast desorption conducting in the gastric fluid lasted shortly and slow desorption occurred in intestinal fluid; pepsin can enhance the release of MMA in gastrointestinal system. While, the acidic condition in gastric fluid is the main factor which causes the release of MMA. The amount of MMA released from CNTs in different fluids follows the order gastric > background (pH = 2.0) > intestinal (fed) > intestinal (fasted) > background (pH = 7.5). These findings in the simulated gastrointestinal system suggest that the release of MMA from CNTs could be promoted by biomacromolecules (such as pepsin and bile salts in digestive tract); thus, the bioavailability of MMA is enhanced.

To determine whether 2-amino-3-methylaminopropanoic acid (BMAA) could be taken up by marine organisms from seawater or their diet mussels Mytilus galloprovincialis, collected from the North Atlantic Portuguese shore, were exposed to seawater doped with BMAA standard (for up to 48 h) or fed with cyanobacteria (for up to 15 days). Mussels were able to uptake BMAA when exposed to seawater. Mussels fed with cyanobacteria Synechocystis salina showed a rise in BMAA concentration during feeding and a decline in concentration during the subsequent depuration period. Cells from the gills and hepatopancreas of mussels fed with S. salina showed lessened metabolic activity in mussels fed for longer periods of time. A hot acidic digestion (considered to account for total BMAA) was compared with a proteolytic digestion, using pepsin, trypsin and chymotrypsin. The latter was able to extract from mussels approximately 30 % of total BMAA. Implications for BMAA trophic transfers in marine ecosystems are discussed.