Global Meta-Analysis and Review of Microplastic in Marine Copepods.

Plastic pollution has spread through all parts of the marine environment, representing a significant threat to species and ecosystems. This study investigates the role of copepods as widespread microplastic reservoirs in the marine environment, by performing, a systematic review, meta-analysis, and semiquantitative analysis of scientific articles focusing on the interaction between copepods and microplastics under field conditions. Our findings indicate that despite uniformly low ingestion of microplastics across different marine layers and geographical areas, with a slight uptake in neustonic copepods, copepods might constitute one of the largest marine microplastic reservoirs. This phenomenon is attributed more to their vast abundance than to average microplastic ingestion values. In this article, a framework for data analysis and reporting is proposed to facilitate future large-scale evaluations and modelling of their extent and impact on plastic and carbon cycles. These insights place copepods at the forefront of the marine plastic cycle, possibly affecting plastic distribution, and bioavailability, thereby opening new pathways for understanding the complex dynamics of microplastics in marine ecosystems.

The hyperbenthic environment: A forgotten habitat for plastic pollution.

This study investigates the abundances and composition of microplastics (MP) among the shallow layers of a coastal Mediterranean Marine Protected Area (Cabrera MPA), seafloor sediments, hyperbenthic environment, and the water column. The mid waters samples were collected mid-way between the sea surface and the seafloor and hyperbenthic samples at the water layer adjacent to the seafloor. Sampling was carried out on patchiness seafloor of Posidonia oceanica meadows. The seafloor sediments showed a mean abundance of 378,769.20 ± 508,109.11 MPs/m3, three orders of magnitude higher than the hyperbenthic (209.17 ± 117.07 MPs/m3), and the mid waters layer (106.48 ± 107.17 MPs/m3). An increasing vertical gradient in MP abundances, mainly composed of fibers was observed. Fibers were made-up mainly of polystyrene (PS, 25 %), expanded polystyrene (EPS, 18 %) and cellulose acetate (CA, 16 %). The results stress the need to increase efforts to find solutions to mitigate fiber pollution in the marine environment.

Breaking the paradigm: Marine sediments hold two-fold microplastics than sea surface waters and are dominated by fibers.

We conducted one of the first studies to integrate the quantification and characterization of microplastics (MPs), including fibers, in different habitats (sea surface, seafloor and beach sediments) of a coastal Mediterranean marine protected area, analyzing their ingestion in several marine species. The objectives of the study were to evaluate the distribution of MPs according to shape and polymer, to assess the contribution of fibers to local plastic pollution and to evaluate their ingestion in fish and invertebrates species that inhabit the study area (Pagrus pagrus, Serranus scriba, Spondyliosoma cantharus, Diplodus vulgaris, Oblada melanura, Holothuria forskalii, Holothuria tubularis, Holothuria polis, Arbacia lixula, Paracentrotus lividus, Modiolus barbatus, Mytilus galloprovincialis and Arca noae). A total of 111 environmental samples were analyzed. The mean abundance of MPs (excluding fibers) quantified in beach sediments (13,418.86 ± 28,787.99 MPs/m2) was two orders of magnitude higher than that found in seafloor sediments (76.92 ± 108.84 MPs/m2), which in turn was two orders of magnitude higher than sea surface samples (0.17 ± 0.39 MPs/m2). The fibers were the most abundant shape of MPs identified in all habitats. Variability in MPs ingestion was detected between species, with ingestion rates ranging from 43 % to 100 % for general MPs and ranging from 7 % to 100 % for fibers. The highest ingestion was observed in Holoturians, representing suitable bioindicators for plastic pollution. The composition of the polymer varies weakly depending on habitats and biota, but the result is strongly correlated with the morphology of the plastic. Fibers were mainly composed of cellulose acetate (29 %), styrofoam of polystyrene (18 %), and filaments, films and fragments of polyethylene and polypropylene. The results highlighted the need to expand integrated approaches to effectively study marine plastic pollution and to undertake efficient actions to limit the input of plastics, particularly fibers, into the marine environment.

Marine plastics in Mediterranean islands: Evaluating the distribution and composition of plastic pollution in the surface waters along four islands of the Western Sea Basin.

To study the spatial distribution of sea surface plastics in marine protected and non-protected areas, 65 sea surface trawls were carried out using a Hydro-bios manta net coupled with a 335-µm mesh. A total of 19 sampling sites along the coastal waters of Mallorca, the "Parque Nacional Marítimo-Terrestre del Archipiélago de Cabrera" and Menorca in the Balearic Islands as well as along coastal waters of The Natural Park of Columbretes Islands (NW Mediterranean Sea) were sampled. A total of 10,637 plastic items were identified and a subset of these items was categorized by shape, color, size and polymer composition. Plastic particles were found at each sampling site and in all samples. No microscale nor mesoscale variability in floating marine plastics abundance (particles/m2) was encountered throughout the study area where similar values were found in protected areas with no local land-based contamination sources, such as Columbretes [0.04 (±0.03) particles/m2], and in high anthropized areas, such as the island of Mallorca [0.04 (±0.07) particles/m2]. However, differences were found in characteristics of plastic items (shape, polymer, and size range), with the protected area of Columbretes characterized by the presence of the highest density of very small plastic items composed mainly of fragments (93%). Quantified plastics from the marine environment were composed mainly of polyethylene (PE, 63.3%), polypropylene (PP; 24.9%), polycarbonate (PC; 4.6%) and polystyrene (PS, 3.3%). The polymer composition showed a homogenous composition between islands and differences were detected only amongst Columbretes and the other islands. Results from this study provide further evidence of the ubiquity of plastics in the marine environment and highlight that remote and protected areas, such as Columbretes, are not exempt from plastic pollution, but receptor areas for small and aged floating plastics composed mainly by fragments, which might have potentially harmful effects on protected ecosystems.

Neustonic microplastics and zooplankton in coastal waters of Cabrera Marine Protected Area (Western Mediterranean Sea).

The high abundance of microplastics in marine environments is becoming a growing threat for Marine Protected Areas (MPAs). Recent studies have identified microplastics (MP) as having multiple effects on biota, and it is of special interest to highlight their effects on zooplanktonic organisms. These organisms play a key role on local food web structures and there is a need to better understand the accumulation of MPs and associated contaminants within the food web. The present study addresses MP distribution and composition within Cabrera MPA as well as its effects on local zooplanktonic community composition. Neustonic microplastic and zooplankton abundance and composition were determined in the shallow coastal waters of Cabrera MPA during the summer season of 2019. Samples were taken using a sea surface manta-net, with a mesh size of 335 µm. MPs were found in all manta trawl samples for a total of 7047 MP with an overall mean abundance (± SD) of 3.52 (± 8.81) items/m3, a value higher than those reported for the majority of Western Mediterranean Sea basins. Qualitative and semi-quantitative analyses of the local zooplankton community suggest that its composition could be sensitive to MP abundances. Although no correlation was found between overall mean abundance of MP and zooplankton, a positive correlation was found between MP abundance and the abundance of the planktonic stage of the foraminifer Rosalina globularis (Tretomphalus). This species seems to be able to use MP for its dispersion. On the other hand, a negative correlation between MP abundance and Copepoda abundance was detected. This work confirms that the coastal zooplankton community composition within the MPA of Cabrera are not exempt from MP pollution and suggests important and complex interactions between MP and zooplankton organisms in coastal waters.

Microplastic intake and enzymatic responses in Mytilus galloprovincialis reared at the vicinities of an aquaculture station.

Aquaculture is a potential source of microplastics (MPs) that could be strong stressors for marine organisms. In this study, we evaluated the effects of MPs derived from aquaculture in antioxidant defences and oxidative stress markers in gills of Mytilus galloprovincialis. Mussels were distributed in three areas with different impacts: inside aquaculture cages, Control 1 (located inside Andratx harbour) and Control 2 (located in a no-anthropized area). Samples were obtained along three different time periods in May (T0), July (T60) and in September (T120). At each sampling period, mussels' biometric measurements were taken, and tissue samples were kept frozen for biochemical determinations and to determine the intake of MPs. An increase in MPs intake was detected throughout the study, and this increase was significantly higher in samples from the aquaculture cages. Similarly, antioxidant enzyme activities (catalase, superoxide dismutase, glutathione reductase and glutathione peroxidase) were significantly higher in samples from cages at T120. Additionally, a similar tendency was observed in glutathione-s-transferase, with a higher activity in the aquaculture cages at T60 and T120. Malondialdehyde and carbonyl protein derivates as a marker of oxidative damage were also measured and samples from aquaculture cages presented higher oxidative stress markers, mainly in T120. In conclusion, living in environments exposed to aquaculture activities at sea may imply a higher intake of MPs which in turn might cause an antioxidant response in M. galloprovincialis which is not enough to avoid oxidative damage.

Impact of the marine litter pollution on the Mediterranean biodiversity: A risk assessment study with focus on the marine protected areas.

In this paper a novel methodology to assess the risk of marine litter (ML) pollution in the Mediterranean Sea is implemented. In this approach, the hazard component is estimated using a state-of-the-art 3D modeling system, which allows the simulation of floating and sinking ML particles; the exposure component is defined from biodiversity estimates; and the vulnerability is related to ML ingestion rates of each species. The results show that the hot-spots for the ML risk concentrate in the coastal regions, and are mainly conditioned by the biodiversity in the region. A dedicated analysis on the marine protected areas shows that the risk therein is controlled by the proximity to ML sources and that their present-day protection levels are not effective in the case of ML pollution. Only a reduction of ML at the sources could reduce the impact of ML pollution in protected areas.

Long-term exposure to virgin and seawater exposed microplastic enriched-diet causes liver oxidative stress and inflammation in gilthead seabream Sparus aurata, Linnaeus 1758.

Plastics accumulation in marine ecosystems has notable ecological implications due to their long persistence, potential ecotoxicity, and ability to adsorb other pollutants or act as vectors of pathogens. The present work aimed to evaluate the physiological response of the gilthead seabream (Sparus aurata) fed for 90 days with a diet enriched with virgin and seawater exposed low-density polyethylene microplastics (LDPE-MPs) (size between 100 and 500 µM), followed by 30 days of depuration, applying oxidative stress and inflammatory markers in liver homogenates. No effects of LDPE-MPs treatments on fish growth were observed throughout this study. A progressive increase in antioxidant enzyme activities was observed throughout the study in both treatments, although this increase was higher in the group treated with seawater exposed MPs. This increase was significantly higher in catalase (CAT), glutathione reductase (GRd), and glutathione-s-transferase (GST) in the seawater exposed MPs group, with respect to the virgin group. In contrast, no significant differences were recorded in superoxide dismutase (SOD) and glutathione peroxidase (GPx) between both groups. Exposure to MPs also caused an increase in the oxidative damage markers (malondialdehyde and carbonyls groups). Myeloperoxidase activity significantly increased because of MPs treatments. After 30 days of depuration, antioxidant, inflammatory enzyme activities and oxidative damage markers returned to values similar to those observed in the control group. In conclusion, MPs exposure induced an increase of antioxidant defences in the liver of S. aurata. However, these elevated antioxidant capabilities were not enough to prevent oxidative damage in the liver since, an increased oxidative damage marker was associated with MPs ingestion. The treatment with seawater exposed MPs caused a more significant antioxidant response (CAT, GRs, and GST). Although after a depuration period of 30 days a tendency to recover the initial values of the biomarkers was observed this does not seem to be time enough for a complete normalization.

Lessons learned from an intercalibration exercise on the quantification and characterisation of microplastic particles in sediment and water samples.

An intercalibration exercise on the characterisation of microplastics in marine sediment and water samples was carried out among five laboratories involved in the implementation of the Marine Strategy Framework Directive (MSFD) in their country. The samples were prepared by mixing cleaned natural sediment and sea water with microplastics sets made of particles of various polymers, shapes and colours. Overall, the errors on total counts were under 25% in absolute value. The risk of non-detection and loss of particles is greater than the risk of contamination during sample analysis. Significant differences are observed among particle types. It appears difficult to obtain reliable and comparable data on the colour of microplastics. A comparison of the errors with regards to the protocols used led to recommend NaCl [1.2 g/cm3] density separation for sediment and one filtering step (200 µm). The operators' experience appears as a key factor for the quality of the results.

Age and growth of the endangered fan mussel Pinna nobilis in the western Mediterranean Sea.

The present work, which is the first comparative study of the growth of the fan mussel Pinna nobilis in the western Mediterranean, encompasses 12 populations of this species living in different environments in France and Spain. Two hundred nine shells were processed and used to obtain growth records from the posterior adductor muscle scar. Size-at-age data were fitted to the Von Bertalanffy growth model. Considerable variability in growth parameters and age was detected among the populations. The results show that the only two fan mussel populations remaining in Spain, which live in an estuary and a coastal lagoon, occupy habitats that are optimal for fast growth, but individuals show low longevity, complicating the long-term conservation of the species. Multivariate analyses groups the populations into three groups (SO, EO and LG), and a general model is proposed for each group; the model can be used as an approximation to calculate the ages of individuals living in similar environments.

Microplastic ingestion by Mullus surmuletus Linnaeus, 1758 fish and its potential for causing oxidative stress.

A total of 417 striped red mullet, Mullus surmuletus, were analyzed to study microplastic ingestion and livers of fish were assessed to study effects of microplastics. Nearly one third (27.30%) of the individuals were quantified to ingest microplastics although there was no evidence of oxidative stress or cellular damage in the liver of fish which had ingested microplastics. A small increase in the activity of glutathione S-transferase (GST) of M. surmuletus was detected which could be suggesting an induction of the detoxification systems but these findings should be tested in laboratory conditions under a controlled diet and known concentration of microplastics. Fish from trammel fisheries, operating closer to land and targeting larger individuals, showed higher mean ingestion values than fish from trawling fisheries, and were related to body size, as microplastics ingested increased with total fish length. Consequently, ingestion values of microplastics were not related to sampling distance from land giving further evidence of the ubiquity of microplastics in the marine environment. Finally, Fourier Transform Infrared Spectroscopy (FTIR) analysis showed that the vast majority of microplastics were filament type and polyethylene terephthalate (PET) was the main identified component.

High levels of microplastic ingestion by the semipelagic fish bogue Boops boops (L.) around the Balearic Islands.

For the first time this study reports on the presence of microplastics (1 nm to <5 mm) in the gastrointestinal tracts of small semipelagic fish (Boops boops) in the Balearic Islands (Mediterranean Sea) from March to May 2014. The results show microplastic ingestion in 68% of full stomach samples with an average of 3.75 items per fish. Only filament type microplastics were observed in B. boops full gastrointestinal tracts. The frequency of occurrence of microplastics was high, with values ranging from 42% to 80%, in comparison to the other ingested items. Spatial variability among locations is high, which suggests that this type of contamination is ubiquitously distributed and originates from multiple sources. The results are important and indirectly provide further evidence of the presence of microplastics, which can be ingested by biota, in the marine environment.

Adapting to the wild: the case of aquaculture-produced and released meagres Argyrosomus regius.

The performance of juvenile Argyrosomus regius released off the coast of Mallorca Island (Balearic Islands, Spain) was assessed by comparing the body condition, stable isotope signature and stomach contents of aquaculture-produced A. regius that had been released, fished and returned by fishermen after spending from a few days to >1 year at liberty with A. regius reared under controlled conditions on two contrasting diets (well-fed and unfed). During the first 40 days of the experiment, the condition index (K(R)) of the returned A. regius and the unfed A. regius followed the same decreasing trend. Thereafter, the K(R) values of the returned A. regius were significantly higher than those of the unfed A. regius but never reached the values of well-fed A. regius. The δ¹³C signal of the returned A. regius clearly increased (in comparison with the well-fed A. regius) after they had spent a few months at liberty. The temporal pattern depicted by the stable isotopes and the most likely prey composition inferred from this pattern strongly suggest a shift in diet. The stomach contents of the returned A. regius that had spent <100 days at liberty consisted almost exclusively of decapods. The diet of the few returned A. regius that had spent >100 days at liberty consisted entirely of fishes. Wild A. regius from the remaining fishery on the Spanish coast exhibited the same ontogenetic diet shift from invertebrates to fishes, but at a smaller size threshold. Overall, the results demonstrated that culture-reared A. regius experience adverse conditions during the first days after release into the wild but that at least some A. regius are able to adapt to the natural environment after a few months at liberty.

Differences in delta13C and delta15N stable isotopes in the pearly razorfish Xyrichtys novacula related to the sex, location and spawning period.

In the present study, Xyrichtys novacula (Labridae) were sampled at five locations around the islands of Ibiza and Formentera (western Mediterranean Sea). Isotopic signatures of delta13C, delta15N and the C:N ratio were analysed in relation to locality, sex and size differences. delta13C and delta15N partitioning was also studied in the reproductive spawning period. There were significant differences in the delta13C signature between localities for both sexes, but not for delta15N. Sex differences were also found with a mean +/-s.e. value of -17.38 +/- 0.06 per thousand delta13C and 8.36 +/- 0.05 per thousand delta(15)N for females and -17.17 +/- 0.07 per thousand delta13C and 8.80 +/- 0.06 per thousand delta15N for males. Increasing total length in both sexes was positively correlated with delta15N enrichment and a significant positive linear regression was established for both variables. During the reproductive spawning period, there were changes in delta13C fractioning with enrichment in postspawning females and males (with respect to prespawning and spawning periods) and delta(15)N impoverishment in postspawning females (with respect to prespawning and spawning periods). Xyrichtys novacula uses local food sources, as confirmed by delta(13)C and delta(15)N, and females and males use different food sources, thus avoiding intraspecific competition. This was confirmed by delta15N enrichment as size increased. Spawning leads to special requirements for gonad maturation, which is reflected in the isotopic signatures for both sexes.

Muscle and scale isotopic offset of three fish species in the Mediterranean Sea: Dentex dentex, Argyrosomus regius and Xyrichtys novacula.

Diet information of organisms was traditionally acquired by the use of lethal techniques (gut content or muscle delta(13)C and delta(15)N isotopic ratios). An increase in the number of isotopic ratio studies and the vulnerability of some species have led to increased use of non-lethal methodologies for conservation purposes. In the present work we have compared the delta(13)C and delta(15)N isotopic signals of muscle and scales of three different fish species in order to test fish-scale sampling as a non-lethal technique in fish trophodynamics. A positive correlation was found between muscle and scales in Argyrosomus regius and Xyrichtys novacula, while Dentex dentex showed no correlation due to the small length range of this species. The isotopic offset (Delta(13)C and Delta(15)N) between muscle and scales was significantly different among species (analysis of variance (ANOVA), p < 0.001) with Tukey's post-hoc HSD indicating a tissue offset difference (p < 0.001) for both stable isotopes in all species, except for Delta(13)C between A. regius and X. novacula and for Delta(15)N between D. dentex and X. novacula (p > 0.05). Mean delta(13)C and delta(15)N values between species showed significant differences (paired t-test, p < 0.01) between muscle and scale with an enrichment for delta(13)C and a depletion for delta(15)N. Spatial differences were found in the Delta(13)C of X. novacula between the studied locations, while no differences were found for Delta(15)N, indicating that non-geographical differences should be considered in the application of scales instead of muscle for (15)N, while for (13)C differences in the geographical isotopic offset should be considered.

Stable isotopes and metal contamination in caged marine mussel Mytilus galloprovincialis.

Metal concentrations and isotopic composition were measured in different tissues of the mussel Mytilus galloprovincialis in waters of the Balearic Islands (Western Mediterranean) in order to assess pollution levels. The isotopic composition was correlated with lead, cadmium, selenium and nickel obtained from the digestive gland and foot of the mussels. Significant negative correlations were found between cadmium, selenium and zinc and the mussel foot, mainly for (13)C. Significant correlations were also found between lead and cadmium and the digestive gland. Pearson correlations indicated that the (13)C isotopic signal in foot is a good proxy for the concentration of metals such as lead, cadmium, selenium and zinc. Similarly, (15)N isotopic signatures in the digestive gland reflected the lead and cadmium concentration.

Assessment of environmental pollution at Balearic Islands applying oxidative stress biomarkers in the mussel Mytilus galloprovincialis.

The antioxidant enzyme response of the mussel Mytilus galloprovincialis to different degree of pollution was investigated. Antioxidant enzyme activities - catalase (CAT), glutathione peroxidase (GSH-PX), glutathione reductase (GR), superoxide dismutase (SOD) - and malondialdehyde (MDA) concentration were measured in gills and digestive glands of mussels. Mussels from the same origin were transplanted along the Balearic coastal waters in eight stations characterized by a different degree of contamination and human impacts. Antioxidant enzyme activities showed an adaptive response to increase the activities in the more polluted areas. CAT, GR and SOD in gills and CAT and GR in digestive gland presented significant differences between polluted and non-polluted stations. No significant differences were observed in MDA concentration indicating that the antioxidant response is capable to avoid the lipid peroxidation. The use of biomarkers such as CAT and GR in gills and digestive glands of the mussel M. galloprovincialis is a good tool to categorize differences between polluted and non-polluted areas.

Organic compounds temporal trends at some invertebrate species from the Balearics, Western Mediterranean.

Concentrations of persistent organic pollutants (POPs) such as hexachlorobenzene (HCB), dichlore diphenyl trichloretane (DDT), polychlorinated biphenyls (PCBs), and gamma-hexachlorocyclohexane (gamma-HCH or lindane) were determined in tissue of marine benthic invertebrates such as Mytilus galloprovincialis, Chamelea gallina, Venus verrucosa, Lithophaga lithophaga and Paracentrotus lividus. Species were selected due to their habitat, trophic level, feeding behaviour and their consumption. Invertebrate species were systematically sampled from December 1996 to December 2005 from several sites along the Balearic Islands. The highest concentrations of PCBs (785ng/g lipid) were found in M. galloprovincialis while the lowest concentrations were found in the sea-urchin P. lividus (193ng/g lipid). Among the 7 PCB quantified congeners the higher values are mainly obtained for CB138 and CB153. All bivalves presented higher PCBs contents than the sea-urchin P. lividus are possibly linked with the bioaccumulation process of POPs throughout the food web and to differential detoxifying mechanisms. The concentration of SigmaDDT exceeds that of HCB and gamma-HCH at all species and sampling stations. DDT concentrations ranged from 0.4ng/g ww at the bivalve C. gallina in 2002, to values of 15.8ng/g ww at the bivalve L. lithophaga in 1998. The values obtained for the organic compounds (HCH, HCB, PCBs, DDT) depend upon the place and year of sampling and are compared to values found by other authors for the mussel M. galloprovincialis in other Mediterranean areas. gamma-HCH and HCB were found in lower concentrations than the other POPs.