Application of non-lethal bioSPME-LC-MS/MS for the detection of human pharmaceuticals in soft corals: A survey at the North Nilandhe atoll (Maldives).

At present the information regarding the occurrence of human pharmaceuticals (PhaCs) in coral reefs and their potential impacts on the associated fauna is limited. To optimize the collection of data in these delicate environments, we employed a solid-phase microextraction (bioSPME) and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) procedure that enabled in vivo determinations in soft corals. Specifically, we researched the antibiotics Ofloxacin Sulfamethoxazole and Clarithromycin, the anti-inflammatory Diclofenac Propyphenazone Ketoprofen and Amisulpride, the neuroactive compounds Gabapentin-lactam, the beta-blocker Metoprolol and the antiepileptic Carbamazepine. Reproducibility was between 2.1% and 9.9% and method detection limits LODs) were between 0.2 and 1.6 ng/g and LOQs between 0.8 and 5.4 mg/g. The method was then applied to establish a baseline for the occurrence of these compounds in the Maldivian archipelago. Colonies of Sarcophyton sp. and Sinularia sp. were sampled along an inner-outer reef transect. Five of the ten targeted PhaCs were identified, and 40% of the surveyed coral colonies showed the occurrence of at least one of the selected compounds. The highest concentrations were found inside the atoll rim. Oxoflacin (9.5 ± 3.9 ng/g) and Ketoprofen (4.5 ± 2.3 ng/g) were the compounds with the highest average concentrations. Outside the atoll rim, only one sample showed contamination levels above the detection limit. No significant differences were highlighted among the two surveyed soft coral species, both in terms of average concentrations and bioconcentration factors (BCFs).

Sublethal effects induced by different plastic nano-sized particles in Daphnia magna at environmentally relevant concentrations.

A growing number of studies have reported the toxic effects of nanoplastics (NPs) on organisms. However, the focus of these studies has almost exclusively been on the use of polystyrene (PS) nanospheres. Herein, we aim to evaluate the sublethal effects on Daphnia magna juveniles of three different NP polymers: PS-NPs with an average size of 200 nm, polyethylene [PE] NPs and polyvinyl chloride [PVC] NPs with a size distribution between 50 and 350 nm and a comparable mean size. For each polymer, five environmentally relevant concentrations were tested (from 2.5 to 250 µg/L) for an exposure time of 48 h. NP effects were assessed at the biochemical level by investigating the amount of reactive oxygen species (ROS) and the activity of the antioxidant enzyme catalase (CAT) and at the behavioral level by evaluating the swimming behavior (distance moved). Our results highlight that exposure to PVC-NPs can have sublethal effects on Daphnia magna at the biochemical and behavioral levels. The potential role of particle size on the measured effects cannot be excluded as PVC and PE showed a wider size range distribution than PS, with particles displaying sizes from 50 to 350 nm. However, we infer that the chemical structure of PVC, which differs from that of PE of the same range size, concurs to explain the observed effects. Consequently, as PS seems not to be the most hazardous polymer, we suggest that the use of data on PS toxicity alone can lead to an underestimation of NP hazards.

Application of marine sponges for biomonitoring active pharmaceutical ingredients (APIs) in coral reefs. Optimization of an SPME and ESI-LC-MS/MS method.

Chemical pollution is a threat to coral reefs. To preserve them, it is crucial to monitor novel contaminants and assess the related risks. The occurrence of active pharmaceutical ingredients (APIs) in coral reefs has been poorly investigated until now. Under this light, we tested the use of the marine sponge Cf. Hyrtios as bio-monitors and conducted a pilot study in the Faafu Atoll (Maldives). Analyses were carried out by in vivo solid-phase microextraction (SPME) and liquid chromatography (LC) electrospray ionization (ESI) tandem mass spectrometry (MS/MS). Twelve APIs were selected for method optimization. Limits of quantitation (LOQs) were in the 0.6 and 2.5 ng/g range, accuracy between 86.5 % and 104.7 %, and precision between 3.0 % and 14.9 %. All the sponges located in the inner reefs resulted contaminated with at least one API. Gabapentin and Carbamazepine displayed the highest detection rates, while Ketoprofen had the highest concentration (up to 15.7 ng/g).

Phthalate levels in common sea anemone Actinia equina and Anemonia viridis: A proxy of short-term microplastic interaction?

Phthalates are widely employed plasticizers blended to plastic polymers that, during plastic aging and weathering are prone to leach in the surrounding environment. Thus, phthalates were proposed to indirectly evaluate MPs contamination in marine environments, with still uncertain and scarce data, particularly for wildlife. This study investigates simultaneously microplastics (MPs) and phthalates (PAEs) occurrence in wild Actinia equina and Anemonia viridis, two common and edible sea anemone species. Both species had a 100 % frequency of MPs occurrence, with similar average concentrations. PAEs were detected in 70 % of samples, with concentrations up to 150 ng/g in A. equina and 144.3 ng/g for A. viridis. MPs and PAEs present in sea anemone tissues appear to reflect seawater plastic contamination conditions in the study area. Given the rapid biodegradation of PAEs, occurrence and concentrations of both these additives and their metabolites could be useful tracers of short-term plastic debris-biota interactions.

Effects of polypropylene nanofibers on soft corals.

Current information regarding the effects of both micro- and nano-plastic debris on coral reefs is limited; especially the toxicity onto corals from nano-plastics originating from secondary sources such as fibers from synthetic fabrics. Within this study, we exposed the alcyonacean coral Pinnigorgia flava to different concentrations of polypropylene secondary nanofibers (0.001, 0.1, 1.0 and 10 mg/L) and then assayed mortality, mucus production, polyps retraction, coral tissue bleaching, and swelling. The assay materials were obtained by artificially weathering non-woven fabrics retrieved from commercially available personal protective equipment. Specifically, polypropylene (PP) nanofibers displaying a hydrodynamic size of 114.7 ± 8.1 nm and a polydispersity index (PDI) of 0.431 were obtained after 180 h exposition in a UV light aging chamber (340 nm at 0.76 Wˑm-2ˑnm-1). After 72 h of PP exposure no mortality was observed but there were evident stress responses from the corals tested. Specifically, the application of nanofibers at different concentrations caused significant differences in mucus production, polyps retraction and coral tissue swelling (ANOVA, p < 0.001, p = 0.015 and p = 0.015, respectively). NOEC (No Observed Effect Concentration) and LOEC (Lowest Observed Effect concentration) at 72 h resulted 0.1 mg/L and 1 mg/L, respectively. Overall, the study indicates that PP secondary nanofibers can cause adverse effects on corals and could potentially act as a stress factor in coral reefs. The generality of the method of producing and assaying the toxicity of secondary nanofibers from synthetic textiles is also discussed.

Plastitar in the Mediterranean Sea: New records and the first geochemical characterization of these novel formations.

A new geological formation consisting of plastic debris admixed to petroleum oil residue, termed "plastitar", has been recently described in the Canary Islands. Here, we report its widespread occurrence across the Mediterranean coast and new insights into its biogeochemical composition. Specifically, we found marked differences in the diagenetic stable indicator profiles, suggesting a heterogeneous seeps provenance. Moreover, the 801 plastic particles found in the 1372 g of tar surveyed, with a maximum concentration of 2.0 items/g, showed interesting patterns in the tar mat, with nurdles predominantly layered in the external of the tar mat and lines in the inner core. Overall, the collected observation suggests that tar entraps plastics through a stepwise process and is a sink for them.

Detection of microplastics and phthalic acid esters in sea urchins from Sardinia (Western Mediterranean Sea).

The occurrence of microplastics (MPs) and phthalic acid esters (PAEs) in wild purple sea urchins (Paracentrotus lividus) of Sardinia (Italy, Western Mediterranean Sea) was surveyed. Specifically, MPs were analyzed in the digestive tract by µFTIR and PAEs in the gonads by SPME-LC-MS/MS. 9 out of 22 specimens resulted contaminated with MPs and 20 displayed levels of PAEs over the quantification limit. A total of 23 MPs were detected with a maximum concentration of 4 microplastics/individual in the commercially undersized specimens. PAEs displayed average concentration of 32 ng/g, σ = 5.3 with maximum value of 77 ng/g. The most abundant congeners were DEHP (17 ng/g, σ = 4.3) and DBP (10 ng/g, σ = 2.5). Statistical analysis showed correlation between DEHP and fiber concentrations and among the concentration of MEP, DEP, DBP and BBzP. Due to local use of sea urchin gonads as gourmet delicacy, the potential human exposition to MPs and PAEs by consumption is also discussed.

Leibniz, the microscope and the concept of preformation.

In recent years a certain emphasis has been put by some scholars on Leibniz's concern about empirical sciences and the relations between such concern and the development of his mature metaphysical system. In this paper I focus on Leibniz's interest for the microscope and the astonishing discoveries that such instrument made possible in the field of the life sciences during the last part of the Seventeenth century. The observation of physical bodies carried out by the "magnifying glasses" revealed a matter swarming everywhere with life and activity, contrary to the cartesian and atomistic view of matter as something sterile and passive. Moreover, the discovery of uncountable complete "animalcula" living in the smallest drop of water provided evidence for the idea of the preformation of every organism. During his lifetime, Leibniz was extremely watchful about the new microscopical discoveries and came into contact with some of the major "observers" of his time, such as Hooke, Leeuwenhoek, Swammerdam and Malpighi. Relying both on some passages in Leibniz's texts and on recent critical studies, I will argue that important aspects of his metaphysics have been strongly affected by the empirical observation of the "invisible world" which the microscope made possible. In the last part of the paper I show how the concept of "preformation", originally drawn from the context of the life sciences, comes to play in Leibniz's philosophy a very general role, going far beyond the scope of biology and shaping important aspects of his overall philosophical system.