Microplastic accumulation and ecological impacts on benthic invertebrates: Insights from a microcosm experiment.

Microplastic (MP) pollution poses a global concern, especially for benthic invertebrates. This one-month study investigated the accumulation of small MP polymers (polypropylene and polyester resin, 3-500 µm, 250 µg L-1) in benthic invertebrates and on one alga species. Results revealed species-specific preferences for MP size and type, driven by ingestion, adhesion, or avoidance behaviours. Polyester resin accumulated in Mytilus galloprovincialis, Chamelea gallina, Hexaplex trunculus, and Paranemonia cinerea, while polypropylene accumulated on Ulva rigida. Over time, MP accumulation decreased in count but not size, averaging 6.2 ± 5.0 particles per individual after a month. MP were mainly found inside of the organisms, especially in the gut, gills, and gonads and externally adherent MP ranged from 11 to 35 % of the total. Biochemical energy assessments after two weeks of MP exposure indicated energy gains for water column species but energy loss for sediment-associated species, highlighting the susceptibility of infaunal benthic communities to MP contamination.

Morphological characterization of the antenna of Torymus sinensis (Hymenoptera: Torymidae) and a comparison within the superfamily Chalcidoidea.

The parasitoid Torymus sinensis (Hymenoptera: Torymidae) has been successfully used in Italy since 2005 for biological control of the invasive cynipid Dryocosmus kuriphilus (Hymenoptera: Cynipidae), highly destructive for the economically relevant Castanea sativa (Fagales: Fagaceae). In order to investigate the morphological aspects related to sensorial behavior, a fine morphology study of the antennae and their sensilla was conducted by scanning electron microscopy on both sexes of T. sinensis. The antennae, composed of a scape, a pedicel and a flagellum with ten flagellomeres, had chaetic sensilla of six subtypes, placoid sensilla of three subtypes, trichoid sensilla, sensilla with a roundish grooved tip, and coeloconic sensilla. The chaetic sensilla of the first three subtypes were found in the scape and in the pedicel, and those of the last three subtypes, together with trichoid, roundish grooved tip and coeloconic sensilla, were found only on flagellomeres. Sexual dimorphism was detected in the morphology of the proper pedicel and the flagellum, and in the presence and distribution of the sensilla and their subtypes. The morphological aspects of the antenna of T. sinensis and of its sensilla were compared with those found in the family Torymidae and in other families of the extremely diverse superfamily Chalcidoidea.

An Overview of Ovarian Calyx Fluid Proteins of Toxoneuron nigriceps (Viereck) (Hymenoptera: Braconidae): An Integrated Transcriptomic and Proteomic Approach.

The larval stages of the tobacco budworm, Heliothis virescens (Fabricius) (Lepidoptera: Noctuidae), are parasitized by the endophagous parasitoid wasp, Toxoneuron nigriceps (Viereck) (Hymenoptera: Braconidae). During the injections of eggs, this parasitoid wasp also injects into the host body the secretion of the venom gland and the calyx fluid, which contains a polydnavirus (T. nigriceps BracoVirus: TnBV) and the Ovarian calyx fluid Proteins (OPs). The effects of the OPs on the host immune system have recently been described. In particular, it has been demonstrated that the OPs cause hemocytes to undergo a number of changes, such as cellular oxidative stress, actin cytoskeleton modifications, vacuolization, and the inhibition of hemocyte encapsulation capacity, which results in both a loss of hemocyte functionality and cell death. In this study, by using a combined transcriptomic and proteomic analysis, the main components of T. nigriceps ovarian calyx fluid proteins were identified and their possible role in the parasitic syndrome was discussed. This study provides useful information to support the analysis of the function of ovarian calyx fluid proteins, to better understand T. nigriceps parasitization success and for a more thorough understanding of the components of ovarian calyx fluid proteins and their potential function in combination with other parasitoid factors.

Additives, plasticizers, small microplastics (<100 µm), and other microlitter components in the gastrointestinal tract of commercial teleost fish: Method of extraction, purification, quantification, and characterization using Micro-FTIR.

One of the aims of this study is the development of a pretreatment method for additives, plasticizers and other components of micro-litter (APFs), and small microplastics (SMPs <100 µm) in the gastrointestinal tract (GIT) of five of the most widely distributed and consumed commercial fish species, Engraulis encrasiculos, Sardina pilchardus, Mullus surmuletus, Solea solea, and Sparus aurata. The second aim was to develop a simultaneous quantification and identification method via Micro-FTIR of APFs and SMPs ingested by these commercial fish species. The distribution of SMPs and APFs is characteristically different for each species investigated. E. encrasiculos and S. pilchardus had a higher weight of SMPs than the other species investigated. Regarding APFs, the highest abundance was observed in E. encrasiculos. This study highlights the importance of studying additives and plasticizers that can be used as efficient proxies of microplastics, as shown by the presence of vulcanizing agents such as Vanax®.

Microplastic accumulation in commercial fish from the Adriatic Sea.

We assessed the presence and concentration of microplastics in fishes from the Northern Adriatic Sea on sale for human consumption. Microplastics were extracted from the digestive tract of 180 specimens, belonging to 6 different species (2 pelagic, and 4 demersal). Microplastics were identified through Nile red staining, and selected particles were analyzed by µ-Raman spectroscopy. The 47.8% of examined fish were found to contain microplastics (233 fragments). The mean concentration of microplastics in the 6 species ranged between 4.11 ± 2.85 items/individual in Adriatic soles, and 1.75 ± 0.71 items/individual in pilchards. Microplastics ranged in size between 0.054 mm and 0.765 mm. Polyethylene and polypropylene were the polymer types identified through µ-Raman spectroscopy. Despite our estimates are underestimations, these results are of particular concern, as the fish analyzed are consumed without being eviscerated, and the microplastics they contain are transferred directly to humans.

Habitat heterogeneity: A confounding factor for the effect of pollutants on macrobenthic community in coastal waters.

Discrepancies were found between Ecological Status of sediments evaluated though the analysis of macrobenthic community and chemical analyses along a heavily trafficked and contaminated maritime seaway in Tyrrhenian Sea. Chemical analyses showed some sites highly contaminated by PAHs and metals with values exceeding thresholds for Good Chemical status and representing potential toxicological risk for benthic animals. Conversely, macrobenthic communities were highly rich and diverse, with Ecological Status (through M-AMBI) Good and High at each site. Sampling depth, total organic carbon, and total nitrogen were the major factors influencing biotic indices (AMBI, M-AMBI, S, H), even if concentrations of PAHs and metals (V, As, and Fe) contributed in explaining a part of indices variability. Habitat heterogeneity of sampled sites is likely acting as a confounding factor for two reasons: (1) high variability of environmental parameters leads to high richness and diversity, to which M-AMBI is sensitive, and (2) environmental parameters explained part of the variability of indices, together with contaminants. Our results suggested the importance of considering natural variability as a fundamental step of environmental impact assessment, for the correct interpretation of biotic indices.

Temporal variation of floatable plastic particles in the largest Italian river, the Po.

We investigated the temporal concentration of floatable plastic particles in the Po river, the largest contributor of freshwater to the Adriatic Sea. Surface waters were sampled in 2019 with a Manta trawl, with almost bimonthly frequency. In total, 5063 plastic particles were collected, 80.6% of which were microplastics (<5 mm). Characterization through FT-IR spectroscopy evidenced 7 polymers, of which polyethylene, polypropylene and polystyrene accounted for 40.5%, 25.7%, and 14.9%, respectively. The highest plastic concentration was recorded in December (3.47 particles m-3; 5.89 mg m-3), while the lowest in August (0.29 ± 0.01 particles m-3; 0.22 ± 0.08 mg m-3). We estimated the annual load of floatable plastic particles carried by the Po river to be 145 tons. Plastic particle load was correlated with the hydraulic regime, and was higher in autumn and spring months.

Types, occurrence and distribution of microplastics in sediments from the northern Tyrrhenian Sea.

This is the first survey to investigate microplastic contamination in the Northern Tyrrhenian seafloor, along a 31 km-long transect from the port of Piombino (Tuscany) to the port of Portoferraio (Elba Island). Plastics extracted from 58 sediment samples were counted and identified by Fourier-transform infrared spectroscopy (FTIR). Plastic pollution occurred in sites closer to ports as well as in offshore stations. Microplastics (1-5 mm) accounted for over 80% of particles. For all samples, the dominant microplastic type was filaments, followed by fragments and films. Six polymers were identified: nylon, polyurethane, polyethylene, and polyethylene terephtalate were the most common. This part of the Tyrrhenian Sea is a busy shipping route connecting the mainland to the Elba Island, with thousands cargo and passenger ships passing by per year. Our data constitute a baseline for microplastic research in the Tyrrhenian seafloor.

A new species of Streblospio (Polychaeta: Spionidae) from the northern Adriatic Sea (Mediterranean Sea).

The present study describes a new species of spionid polychaete, Streblospio eridani n. sp., from the Italian coasts of the Northern Adriatic Sea (Mediterranean Sea). This new spionid species was recorded from shallow lagoon and marine habitats: the soft bottoms of the Sacca di Goro (Po River Delta), on October 2017, and the Lido di Dante (Emilia Romagna), between 2016 and 2017. The re-assessment of spionid specimens from other lagoon systems (the Valli di Comacchio and the Valle di Gorino) led us to recognize the presence of this species in the northern Adriatic Sea since 2009. Streblospio eridani n. sp. had low occurrence and density at the marine site Lido di Dante and at the lagoon sites Valli di Comacchio and Valle di Gorino; whereas it reached the highest density of 21,213.3 ind. m-2 at the Sacca di Goro.  Morphologically this new species is characterized by dorsal (occipital) papilla on the first chaetiger, sabre chaetae and hooded hooks first appearing on chaetigers 7, hooks with 4-5 pairs of small secondary teeth, pygidium with ventral lappets, and brooding branchiate structures between chaetigers 13 and 28. In the phylogenetic reconstruction, based on the mitochondrial cytochrome c oxidase subunit I (COI) gene, the Adriatic taxon results clearly indicate that this is distinct from the other species of the genus and formed a well-supported clade with other Streblospio sp. specimens from India. Nucleotide divergences calculated between the Adriatic specimens and the other Streblospio species are higher than the intraspecific range reported for the genus and support the description of a new species. Morphological characters important for differentiation of the new species herein described from congeneric species are discussed and an updated key for Streblospio species is provided.

Microplastic accumulation in benthic invertebrates in Terra Nova Bay (Ross Sea, Antarctica).

Microplastic contamination of the benthic invertebrate fauna in Terra Nova Bay (Ross Sea, Antarctica) was determined. Twelve macrobenthic species, characterized by different feeding strategies, were selected at 3 sampling sites at increasing distance from the Italian Scientific Base (Mario Zucchelli, Camp Icarus, Adelie Cove). The 83% of the analyzed macrobenthic species contained microplastics (0.01-3.29 items mg-1). The size of the particles, measured by Feret diameter, ranged from 33 to 1000 µm with the highest relative abundance between 50 and 100 µm. Filter-feeders and grazers displayed values of microplastic contamination from 3 to 5 times higher than omnivores and predators, leading to the hypothesis that there is no evident bioaccumulation through the food web. The prevalent polymers identified by micro-FTIR were nylon (86%) and polyethylene (5%); other polymers identified in Antarctic benthos were polytetrafluoroethylene, polyoxymethylene, phenolic resin, polypropylene, polystyrene resin and XT polymer.

Adverse effects of plastic ingestion on the Mediterranean small-spotted catshark (Scyliorhinus canicula).

Plastics are widely diffused in the oceans and their ingestion by marine organisms is raising concern for potentially adverse effects. The risk of harmful interactions with marine plastic pollution depends on the biology of the species as well as the distribution and abundance of the different plastic types. The aim of this study was to assess the occurrence of plastic ingestion by the small-spotted catshark (Scyliorhinus canicula), one of the most abundant elasmobranchs in the Mediterranean Sea. The expression levels of genes indicative of total immune system function were analyzed to gather preliminary data for further investigation of any potential correlations between plastic presence and immune activation. One hundred catsharks were collected during the Spring 2018 in two geographic locations in the southern region of the central Mediterranean Sea: 1) near Mazara del Vallo, SW Sicily and 2) near Lampedusa island, Italy's southernmost. Standard measurements were recorded for each specimen and its organs and sex was determined. The gastrointestinal tract (GIT) was preserved for plastic detection and identification. Where present, plastics (macro- and micro-) were characterized in terms of size, shape and polymer typology through microscopy and µ-Raman spectroscopy. Spleen from a subset of thirty samples was preserved for RNA extraction, then used to quantify by real time PCR the transcripts of T cell receptor beta (TCRB), T cell receptor delta (TCRD) and IgM genes. The results indicated that ingestion of plastic is widespread, with microplastics (MP, from 1 µm to <1 mm) abundantly present in nearly all samples and macroplasticplastic (MaP, > 1 cm) in approximately 18% of the specimens collected. A significant increase in the expression of TCRB, TCRD and IgM was observed in the spleen of MaP + specimens from Mazara del Vallo waters, in parallel with 67% increase in liver weight. While the presence of MP alone is not enough to induce a strong activation of the immunity, some type of plastics falling into the MaP category may be more toxic than others and crucial in the activation of the immune response. The results of this study represent a first evidence that plastic pollution represents an emerging threat to S. canicula, the Mediterranean food web and human consumers.

Hazardous effects of silver nanoparticles for primary producers in transitional water systems: The case of the seaweed Ulva rigida C. Agardh.

The acute toxicity of citrate capped silver nanoparticles (AgNP) and silver nitrate was evaluated on the marine macroalga Ulva rigida C. Agardh (1823). Silver bioaccumulation, ultrastructural chloroplast damages verified by TEM microscopy, inhibition of primary production, neutral lipid production and oxidative stress were observed after 24 h of exposure to AgNP. The toxic effects of silver nitrate in artificial seawater started from a concentration of 0.05 ppm and was more toxic than AgNP that produced effects from a concentration of 0.1 ppm. However only AgNP induced lipid peroxidation in U. rigida. The addition of natural organic and inorganic ligands, represented by transparent exopolymer particles (TEP) and clay, drastically reduced AgNP acute toxicity in a ratio AgNP:ligand of 1:100 and 1:200, respectively. The findings suggest a marked toxicity of Ag on marine macroalgae which however should be mitigated by the high natural ligand concentrations of the transitional environments.

The difficulty of disentangling natural from anthropogenic forcing factors makes the evaluation of ecological quality problematic: A case study from Adriatic lagoons.

The complex and dynamic nature of transitional ecosystems pose problems for the assessment of the Ecological Quality Status required by the European Water Framework Directive (WFD; 2000/60/EC). In six Adriatic lagoons, Ecological Quality Status was studied by comparing a biotic index based on macrophytes (MaQI), and three indices based on invertebrates (M-AMBI, M-bAMBI, and ISD). Ecological Status evaluated though MaQI and ISD resulted in quite degraded ecosystems (moderate/poor/bad), with only opportunistic algae and macrobenthic communities dominated by small size classes. Those results were supported by physico-chemical parameters, indicating high nutrients inputs, and anthropogenic pressures related with agriculture and fishery activities. Ecological Status obtained with M-AMBI and M-bAMBI was higher, with some sites reaching even the "good" status. The best response to anthropogenic pressures, in terms of a pressure index, was obtained by M-AMBI and M-bAMBI. Nevertheless, the response of used metrics (such as AMBI and bAMBI) to environmental variables not related to anthropogenic impact, and the high heterogeneity of physical-chemical conditions within lagoons, represent potential problems for the correct evaluation of Ecological Status of transitional waters. When different metrics give different responses it becomes a problem for managers who cannot easily make a decision on the remedial measures. The disagreement among indices arose because of the different response of biological elements to different stressors, and because the different indices based on macroinvertebrates focused on different aspects of the community, providing complementary information. So urge the need to find alternative approaches for a correct assessment of Ecological Status, with the combination of different biological elements, and considering the development of new indices (e.g. M-bAMBI) or refinement of the existing ones.

Species-Area Relationship (SAR) models as tools for estimating faunal biodiversity associated with habitat builder species in sensitive areas: the case of the Mediterranean stony coral (Cladocora caespitosa).

Biodiversity associated with the Mediterranean stony coral Cladocora caespitosa (Linnaeus, 1767) was investigated at three levels: "microscale", focused on macrobenthic invertebrates within colonies; "mesoscale", focused on epibenthic megafauna among colonies; "macroscale", focused on associated ichthyofauna. The aim was to quantify associated diversity in terms of species richness, testing the efficiency of colony size (surface covered by a single colony) for the "microscale", and colony density or total coral coverage for "meso-" and "macroscale" as predictors and the consistency of models based on Species-Area Relationship (SAR) for those estimations. At level of "microscale", colony size was a good predictor, with richness of invertebrates increasing with the increasing of surface covered by each colony of C. caespitosa, following Arrhenius model. At levels of "mesoscale" and "macroscale", richness of epibenthic megafauna and fish were not related neither to colony density nor total coral coverage, but to sampled area, and frequency-based estimates of richness were used. The importance of C. caespitosa varied according to the investigation level, with most of taxa richness detected at the level of "microscale".

When the levee breaks: Effects of flood on offshore water contamination and benthic community in the Mediterranean (Ionian Sea).

In the last few years extreme weather events, including changes to storm frequency and intensity, have increased across all continents. In this note we assessed, for the first time in the Mediterranean Sea, the impact of a violent storm and consequent flood on offshore water contamination and benthic community along the Calabrian coast (Ionian Sea). Three sites (at 500, 1000, and 2000 m off the coast) were sampled along three parallel transects in 2013, 2014 (before), and 2015 (after the flood). After the flood, metals (especially Al, CrVI, Ni, Cu, Zn) in the water column increased in concentration. The flood affected the structure of the benthic community, causing a decrease of diversity, the dominance of few opportunistic species, and the decrease of M-AMBI values.

The efficiency of Taxonomic Sufficiency for identification of spatial patterns at different scales in transitional waters.

Taxonomic Sufficiency (TS), the use of coarser taxonomic resolution in monitoring plans, has been receiving increasing attention in last years. A comprehensive dataset of macrobenthos from 18 Italian lagoons in a range of different latitude, typology, salinity and surface area, was analysed in order to test the efficiency of TS, in terms of correlation between patterns at level of species and patterns resulting from different levels of taxonomic aggregation. First, TS was applied on a range of univariate indices, providing complementary information on macrobenthic community, in order to test the efficiency, in a contest of different taxonomic composition, and different number of lower taxa belonging to the same higher taxon in each lagoon. Then, TS was applied on multivariate analyses, in order to test whether the efficiency changes between two different scales: local (comparison of sites nested within each lagoon) and regional (comparison among lagoons), and with different data transformation. The patterns resulting from univariate indices and multivariate analyses, at both local and regional scales, were retained till family level, despite the different levels of taxonomic composition and different number of lower taxa belonging to the same higher taxon of different lagoons. Nevertheless, the correlation values among matrices and the effect of data transformation differed between regional and local scales. Our results support the efficiency of TS until family level, but at the same time underline the need of scale- and region-specific baseline knowledge prior application of TS in lagoons.

Heavy metals, OCPs, PAHs, and PCDD/Fs contamination in surface sediments of a coastal lagoon (Valli di Comacchio, NW Adriatic, Italy): Long term trend (2002-2013) and effect on benthic community.

Surface sediments contamination (heavy metals, organochlorine pesticides, polycyclic aromatic hydrocarbons, polychlorinated dibenzodioxins and dibenzofurans) and benthic community of the Valli di Comacchio, were analysed from 2002 to 2013. Along the studied period, most of analysed pollutants did not exceed thresholds for Good chemical status of sediments. In 2008, a peak of contamination was observed, with total PAHs and OCPs exceeding their threshold. Considering metals, Ni and Cr exceeded their thresholds, but not the background levels for the area, suggesting natural enrichment; conversely Pb exceeded background levels for the area in many samples, but it exceeded its sediment quality thresholds only in few samples (2006, 2009, 2011). Conversely, the ecological status evaluated though AMBI and M-AMBI, was below the Good/Moderated thresholds established by WFD in most of the studied years. In the 11 yrs-study period, the benthic community showed a very weak response to chemical contamination.

Long-term variability of macrobenthic community in a shallow coastal lagoon (Valli di Comacchio, northern Adriatic): Is community resistant to climate changes?

A time series data of macrobenthic invertebrates of Valli di Comacchio lagoon (northern Adriatic) from 1996 to 2015, was analysed using Biological Traits Analysis, structural indices, AMBI and M-AMBI indices, with a twofold aim to: firstly, test the resilience of the system, and, secondly, test the influence of climate changes, in terms of temperature and precipitation pattern, on macrobenthic dynamics. Along the studied period macrobenthic community showed marked fluctuations, in terms of richness, diversity, biological traits and ecological groups, which could be related with environmental instability of the lagoon. At the same time, a general tendency towards a deterioration of ecological condition of the lagoon was observed, with a general decrease in species richness, diversity, percentage of sensitive species, and a general increase in the proportion of the more opportunistic trait modalities, such as deposit feeders, burrowing, infaunal and short living animals. Increasing yearly temperature explained only a small part of the variability of macrobenthic community, in terms of biological traits and diversity indices, and this was likely due to the effect of natural fluctuations of environmental parameters and anthropogenic disturbance. Nevertheless, all metrics used are consistent in identifying the response of benthic community to a severe disturbance, likely related with the summer heatwave in 2003. Less marked signs of disturbance were observed also in relations to the thermal anomaly of 2012. Biological Traits Analysis combined with more classical structural and ecological indices, proved to be efficient in identifying temporal changes of the community. Our results suggest that the expected increase in frequency, magnitude and duration of heatwaves could pose serious threat to the resilience capacity of lagoonal macrobenthic community.

Comparison of benthic indices for the evaluation of ecological status of three Slovenian transitional water bodies (northern Adriatic).

Benthic indicators are important tools for the classification of coastal and transitional water bodies. The aim of the work was to assess for the first time the Environmental Status (ES) of Slovenian transitional waters, comparing the following biotic indices: richness, Shannon-Weaver diversity, AMBI, M-AMBI, BENTIX and BITS indices. A total of 13 stations were sampled with a Van Veen grab, in three ecosystems in the northern Adriatic. Samples were sieved and sorted, invertebrates identified and counted. The anthropogenic impact was estimated with professional judgement. Richness and diversity showed a good response to anthropogenic pressure. Conversely, indices based on sensitivity/tolerance groups did not showed a clear distinction between more and less impacted ecosystems. In particular BENTIX underestimated the ES, while with BITS there was a overestimation. The best evaluation was obtained with M-AMBI, because even if based on a sensitivity/tolerance approach, it considered also the structural aspect of the community.