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).

Physical and cellular impact of environmentally relevant microplastic exposure on thermally challenged Pocillopora damicornis (Cnidaria, Scleractinia).

Microplastic pollution is an increasing threat to coral reefs, which are already strongly challenged by climate change-related heat stress. Although it is known that scleractinian corals can ingest microplastic, little is known about their egestion and how microplastic exposure may impair corals at physiological and cellular levels. In addition, the effects of microplastic pollution at current environmental concentration have been little investigated to date, particularly in corals already impacted by heat stress. In this study, the combined effects of these environmental threats on Pocillopora damicornis were investigated from a physical and cellular perspective. Colonies were exposed to three concentrations of polyethylene microplastic beads (no microplastic beads: [No MP], 1 mg/L: [Low MP]; 10 mg/L: [High MP]), and two different temperatures (25 °C and 30 °C) for 72 h. No visual signs of stress in corals, such as abnormal mucus production and polyp extroflection, were recorded. At [Low MP], beads adhered to colonies were ingested but were also egested. Moreover, thermally stressed colonies showed a lower adhesion and higher egestion of microplastic beads. Coral bleaching was observed with an increase in temperature and microplastic bead concentration, as indicated by a general decrease in chlorophyll concentration and Symbiodiniaceae density. An increase in lipid peroxidation was measured in colonies exposed to [Low MP] and [High MP] and an up-regulation of stress response gene hsp70 was observed due to the synergistic interaction of both stressors. Overall, our findings showed that heat stress still represents the main threat to P. damicornis, while the effect of microplastics on coral health and physiology may be minor, especially at control temperature. However, microplastics could exacerbate the effect of thermal stress on cellular homeostasis, even at [Low MP]. While reducing ocean warming is critical for preserving coral reefs, effective management of emerging threats like microplastic pollution is equally essential.

QUIN 2.0 - new release of the QUaternary fault strain INdicators database from the Southern Apennines of Italy.

QUIN database integrates and organizes structural-geological information from published and unpublished sources to constrain deformation in seismotectonic studies. The initial release, QUIN1.0, comprised 3,339 Fault Striation Pairs, mapped on 445 sites exposed along the Quaternary faults of central Italy. The present Data Descriptor introduces the QUIN 2.0 release, which includes 4,297 Fault Striation Pairs on 738 Structural Sites from southern Italy. The newly investigated faults span ~500 km along the Apennines chain, with strikes transitioning from ~SE to ~SW and comprehensively details Fault Striation Pairs' location, attitude, kinematics, and deformation axes. Additionally, it offers a shapefile of the fault traces hosting the data. The QUIN 2.0 release offers a significant geographic extension to the QUIN 1.0, with comprehensive description of local geometric-kinematic complexities of the regional pattern. The QUIN data may be especially relevant for constraining intra-Apennine potential seismogenic deformation patterns, where earthquake data only offer scattered or incomplete information. QUIN's data will support studies aimed at enhancing geological understanding, hazard assessment and comprehension of fault rupture propagation and barriers.

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.

Systematics and character evolution of capitate hydrozoans.

Capitate hydrozoans are a morphologically and ecologically diverse hydrozoan suborder, currently including about 200 species. Being grouped in two clades, Corynida and Zancleida, these hydrozoans still show a number of taxonomic uncertainties at the species, genus and family levels. Many Capitata species established symbiotic relationships with other benthic organisms, including bryozoans, other cnidarians, molluscs and poriferans, as well as with planktonic dinoflagellates for mixotrophic relationships and with bacteria for thiotrophic ectosymbioses. Our study aimed at providing an updated and comprehensive phylogeny reconstruction of the suborder, at modelling the evolution of selected morphological and ecological characters, and at testing evolutionary relationships between the symbiotic lifestyle and the other characters, by integrating taxonomic, ecological and evolutionary data. The phylogenetic hypotheses here presented shed light on the evolutionary relationships within Capitata, with most families and genera being recovered as monophyletic. The genus Zanclea and family Zancleidae, however, were divided into four divergent clades, requiring the establishment of the new genus Apatizanclea and the new combinations for species in Zanclea and Halocoryne genera. The ancestral state reconstructions revealed that symbiosis arose multiple times in the evolutionary history of the Capitata, and that homoplasy is a common phenomenon in the group. Correlations were found between the evolution of symbiosis and morphological characters, such as the perisarc. Overall, our results highlighted that the use of genetic data and a complete knowledge of the life cycles are strongly needed to disentangle taxonomic and systematic issues in capitate hydrozoans. Finally, the colonization of tropical habitat appears to have influenced the evolution of a symbiotic lifestyle, playing important roles in the evolution of the group.

First record of the bull shark Carcharhinus leucas (Valenciennes, 1839) from the Maldivian archipelago, central Indian Ocean.

Verified records of the bull shark Carcharhinus leucas are lacking in the Maldives. This study provides the first confirmed evidence of 23 sightings observed from 2013 to 2023 in the central and southern atolls of this archipelago. Most of the sightings occurred in close proximity to inhabited areas, where food waste is often discarded into the water, or in several dive sites, suggesting the presence of this species in different locations around central and southern atolls. Although further research is required to fully investigate the C. leucas population in the Maldives, this report documents and confirms its presence in this region.

Biodegradable Zein-Based Biocomposite Films for Underwater Delivery of Curcumin Reduce Thermal Stress Effects in Corals.

Massive coral bleaching episodes induced by thermal stress are one of the first causes of coral death worldwide. Overproduction of reactive oxygen species (ROS) has been identified as one of the potential causes of symbiosis breakdown between polyps and algae in corals during extreme heat wave events. Here, we propose a new strategy for mitigating heat effects by delivering underwater an antioxidant to the corals. We fabricated zein/polyvinylpyrrolidone (PVP)-based biocomposite films laden with the strong and natural antioxidant curcumin as an advanced coral bleaching remediation tool. Biocomposites' mechanical, water contact angle (WCA), swelling, and release properties can be tuned thanks to different supramolecular rearrangements that occur by varying the zein/PVP weight ratio. Following immersion in seawater, the biocomposites became soft hydrogels that did not affect the coral's health in the short (24 h) and long periods (15 days). Laboratory bleaching experiments at 29 and 33 °C showed that coral colonies of Stylophora pistillata coated with the biocomposites had ameliorated conditions in terms of morphological aspects, chlorophyll content, and enzymatic activity compared to untreated colonies and did not bleach. Finally, biochemical oxygen demand (BOD) confirmed the full biodegradability of the biocomposites, showing a low potential environmental impact in the case of open-field application. These insights may pave the way for new frontiers in mitigating extreme coral bleaching events by combining natural antioxidants and biocomposites.

Bioconcentration and cellular effects of emerging contaminants in sponges from Maldivian coral reefs: A managing tool for sustainable tourism.

Tourism is the main income source for the Maldives, but concurrently, it represents a growing threat to its marine ecosystem. Here, we monitored the bioaccumulation of 15 emerging contaminants (ECs) in the Maldivian reef sponges Spheciospongia vagabunda collected in two resort islands (Athuruga and Thudufushi, Ari Atoll) and an inhabited island (Magoodhoo, Faafu Atoll), and we analysed their impact on different sponge cellular stress biomarkers. Caffeine and the insect repellent DEET were detected in sponges of all the islands, whereas the antibiotic erythromycin and the UV filter 4-methylbenzylidene camphor were found in resort islands only. Although concentrations were approximately a few ng/g d.w., we quantified various induced cellular effects, in particular an increase of the levels of the enzyme glutathione S-transferase involved in cell detoxification. Our results highlight the importance to increase awareness on ECs pollution, promoting the use of more environmental friendly products to achieving the sustainable development goals.

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.

Playing "hide and seek" with the Mediterranean monk seal: a citizen science dataset reveals its distribution from molecular traces (eDNA).

Animal conservation relies on assessing the distribution and habitat use of species, but for endangered/elusive animals this can prove difficult. The Monk Seal, Monachus monachus, is one of the world's most endangered species of pinniped, and the only one endemic to the Mediterranean Sea. During recent decades, direct observations have been few and scattered, making it difficult to determine its distribution away from the Aegean Sea (core distribution area of the post-decline relict population). This study relies on environmental DNA (eDNA) analysis to detect the presence of the Monk Seal in 135 samples collected in 120 locations of the central/western Mediterranean Sea, spanning about 1500 km longitudinally and 1000 km latitudinally. A recently described species-specific qPCR assay was used on marine-water samples, mostly collected during 2021 by a Citizen Science (CS) project. Positive detections occurred throughout the longitudinal range, including the westernmost surveyed area (Balearic archipelago). The distribution of the positive detections indicated six "hotspots", mostly overlapping with historical Monk Seal sites, suggesting that habitat-specific characteristics play a fundamental role. We applied single-season occupancy models to correct for detection probability and to assess the importance of site-specific characteristics. The distance from small islets and protected (or access-restricted) areas was correlated negatively with the detection probability. This novel molecular approach, applied here for the first time in an extensive CS study, proved its potential as a tool for monitoring the distribution of this endangered/elusive species.

DNA metabarcoding unveils the effects of habitat fragmentation on pollinator diversity, plant-pollinator interactions, and pollination efficiency in Maldive islands.

Habitat fragmentation affects biodiversity, but with unclear effects on pollinators and their interactions with plants in anthropized landscapes. Islands could serve as open air laboratories, suitable to disentangle how land-use alteration impacts pollination ecology. In Maldive islands we investigated how pollinator richness, plant-pollinator interactions and pollination efficiency are influenced by the green area fragmentation (i.e., gardens and semi-natural patches). Moreover, we considered the mediating role of pollinator body size and the plant trait of being invasive in shaping interactions. To do this, we surveyed pollinator insects from 11 islands representing a gradient of green area fragmentation. A DNA metabarcoding approach was adopted to identify the pollen transported by pollinators and characterize the plant-pollinator interactions. We found that intermediate levels of green area fragmentation characterized pollinator communities and increased their species richness, while decreasing interaction network complexity. Invasive plants were more frequently found on pollinator bodies than native or exotic noninvasive ones, indicating a concerningly higher potential for pollen dispersal and reproduction of the former ones. Intriguingly, pollinator body size mediated the effect of landscape alteration on interactions, as only the largest bees expanded the foraging diet in terms of plant richness in the transported pollen at increasing fragmentation. In parallel, the pollination efficiency increased with pollinator species richness in two sentinel plants. This study shows that moderate landscape fragmentation of green areas shapes many aspects of the pollination ecosystem service, where despite interactions being less complex and mediated by pollinator body size, pollinator insect biodiversity and potential plant reproduction are supported.

Natural Products from Mangroves: An Overview of the Anticancer Potential of Avicennia marina.

Exploring the potential of natural extracts for pharmaceutical applications in the treatment of different diseases is an emerging field of medical research, owing to the tremendous advantages that they can offer. These include compound sustainability due to the natural origin and virtually unlimited availability. In addition, they contribute to promoting the countries in which they are extracted and manufactured. For this reason, wild active compounds derived from plants are attracting increasing interest due to their beneficial properties. Among them, Avicennia marina has been recently recognized as a potential source of natural substances with therapeutic activities for anti-cancer treatment. A. marina beneficially supplies different chemical compounds, including cyclic triterpenoids, flavonoids, iridoids, naphtaquinones, polyphenols, polysaccharides, and steroids, most of them exhibiting potent antitumor activity. The in vivo and in vitro studies on different models of solid tumors demonstrated its dose-dependent activity. Moreover, the possibility to formulate the A. marina extracted molecules in nanoparticles allowed researchers to ameliorate the therapeutic outcome of treatments exploiting improved selectivity toward cancer cells, thus reducing the side effects due to nonspecific spread.

Effects of the COVID-19 lockdowns on the management of coral restoration projects.

Coral restoration initiatives are gaining significant momentum in a global effort to enhance the recovery of degraded coral reefs. However, the implementation and upkeep of coral nurseries are particularly demanding, so that unforeseen breaks in maintenance operations might jeopardize well-established projects. In the last 2 years, the COVID-19 pandemic has resulted in a temporary yet prolonged abandonment of several coral gardening infrastructures worldwide, including remote localities. Here we provide a first assessment of the potential impacts of monitoring and maintenance breakdown in a suite of coral restoration projects (based on floating rope nurseries) in Colombia, Seychelles, and Maldives. Our study comprises nine nurseries from six locations, hosting a total of 3,554 fragments belonging to three coral genera, that were left unsupervised for a period spanning from 29 to 61 weeks. Floating nursery structures experienced various levels of damage, and total fragment survival spanned from 40 to 95% among projects, with Pocillopora showing the highest survival rate in all locations present. Overall, our study shows that, under certain conditions, abandoned coral nurseries can remain functional for several months without suffering critical failure from biofouling and hydrodynamism. Still, even where gardening infrastructures were only marginally affected, the unavoidable interruptions in data collection have slowed down ongoing project progress, diminishing previous investments and reducing future funding opportunities. These results highlight the need to increase the resilience and self-sufficiency of coral restoration projects, so that the next global lockdown will not further shrink the increasing efforts to prevent coral reefs from disappearing.

First detection of microplastics in reef-building corals from a Maldivian atoll.

The presence of microplastics in the world's oceans and their effects on marine habitats are highly concerning. As suspension-feeders, corals are very exposed to microplastics, compromising the health of coral reef ecosystems. In this study we surveyed for the first time the presence of microplastics in Maldivian reef-building corals. Aiming to determine the influence of exposure and depth on microplastic distribution, analyses were carried out on 38 individuals belonging to three different species. 58% of the investigated colonies resulted contaminated with particles within the 25-150 µm size range. The maximum concentration was encountered in a Pocillopora verrucosa colony sampled from a shallow inner reef (8.9 particles/g of coral). No significant differences in microplastic concentration were observed between different depths, exposures, sites and species. Overall, this study confirmed microplastic presence in coral reefs of the Maldivian archipelago including foundation species.

Phthalates bioconcentration in the soft corals: Inter- and intra- species differences and ecological aspects.

The bioconcentration of dimethyl phthalate (DMP) diethyl phthalate (DEP) dibutyl phthalate (DBP) butyl benzyl phthalate (BBzP), di-(2-ethy hexyl) phthalates (DEHP), mono-butyl phthalate (MBP), mono-benzyl phthalate (MBzP), mono-(2-ethy hexyl) phthalate (MEHP) in the soft corals Coelogorgia palmosa, Sinularia sp., Sarcophyton glaucum, and Lobophytum sp. was investigated. Specimens were cultured in a microcosm environment built-up at the Genova Aquarium and analyses were carried out by in vivo SPME-LC-MS/MS. The distributions of the phthalates among the four surveyed species resulted significantly different. Calculated bioconcentration factors (BCFs) showed values spanning over two orders of magnitude, from a minimum of log10 BCFDEP = 1.0 in Sarcophyton glaucum to a maximum of log10 BCFDBP = 3,9 calculated for Coelogorgia palmosa. Moreover, the calculated BCFs of the long chain phthalates resulted up to three orders of magnitude lower than theoretically predicted (from logKow), whereas BCF of short chain phthalates resulted higher. This, together with the detection of phthalic acid monoesters, suggests the presence of species-specific different metabolic transformation among the surveyed soft coral species that involve DEHP.

Comparing different farming habitats for mid-water rope nurseries to advance coral restoration efforts in the Maldives.

The need for comprehensive and effective coral restoration projects, as part of a broader conservation management strategy, is accelerating in the face of coral reef ecosystem decline. This study aims to expand the currently limited knowledge base for restoration techniques in the Maldives by testing the performance of mid-water rope nurseries in a lagoon and a reef habitat. We examined whether different coral farming habitats impacted fragment survival, health and growth of two coral genera and how the occurrence of mutualistic fauna, predation and disease influenced coral rearing success. Two nurseries were stocked with a total of 448 Pocillopora verrucosa and 96 Acropora spp. fragments, divided into different groups (four Pocillopora groups: lagoon nursery at 5 m; reef nursery at 5, 10 and 15 m; two Acropora groups: lagoon nursery at 5 m and reef nursery at 5 m). Eight fragment replicates from the same donor colony (Pocillopora genets: N = 14, Acropora genets N = 6) were used in each group and monitored for one year. Our results show that fragment survival was high in both farming habitats (>90%), with P. verrucosa surviving significantly better in the lagoon and Acropora spp. surviving and growing significantly faster in the reef nursery. P. verrucosa growth rates were similar between reef and lagoon habitat. Different rearing depths in the reef nursery had no impact on the survival of P. verrucosa but coral growth decreased considerably with depth, reducing fragments' ecological volume augmentation and growth rates by almost half from 5 to 15 m depth. Further, higher fish predation rates on fragments were recorded on the reef, which did not impact overall nursery performance. Mutualistic fauna, which correlated positively with fragment survival, was more frequently observed in the lagoon nursery. The occurrence of disease was noted in both habitats, even though implications for fragment health were more severe in the lagoon. Overall, our study demonstrates that lagoon and reef nurseries are suitable for rearing large numbers of coral fragments for transplantation. Nevertheless, we recommend considering the specific environmental conditions of the farming habitat, in particular water quality and year-round accessibility, in each case and to adjust the coral farming strategy accordingly. We hope that this novel research encourages the increased application of mid-water rope nurseries for 'coral gardening' to advance coral reef recovery and climate resilience in the Maldives.

Cytotoxic Compounds from Alcyoniidae: An Overview of the Last 30 Years.

The octocoral family Alcyoniidae represents a rich source of bioactive substances with intriguing and unique structural features. This review aims to provide an updated overview of the compounds isolated from Alcyoniidae and displaying potential cytotoxic activity. In order to allow a better comparison among the bioactive compounds, we focused on molecules evaluated in vitro by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, by far the most widely used method to analyze cell proliferation and viability. Specifically, we surveyed the last thirty years of research, finding 153 papers reporting on 344 compounds with proven cytotoxicity. The data were organized in tables to provide a ranking of the most active compounds, to be exploited for the selection of the most promising candidates for further screening and pre-clinical evaluation as anti-cancer agents. Specifically, we found that (22S,24S)-24-methyl-22,25-epoxyfurost-5-ene-3ß,20ß-diol (16), 3ß,11-dihydroxy-24-methylene-9,11-secocholestan-5-en-9-one (23), (24S)-ergostane-3ß,5α,6ß,25 tetraol (146), sinulerectadione (227), sinulerectol C (229), and cladieunicellin I (277) exhibited stronger cytotoxicity than their respective positive control and that their mechanism of action has not yet been further investigated.

Detection of plastic particles in marine sponges by a combined infrared micro-spectroscopy and pyrolysis-gas chromatography-mass spectrometry approach.

Plastic pollution threatens the marine environment, especially due to the adverse effects caused by micro and nano particles interacting with the marine biota. In order to provide reliable data regarding micro and nanoplastic contamination and the related impacts, efficient analytical solutions are needed. We developed a new analysis workflow that uses marine sponges to monitor plastic pollution by characterizing the plastic particles accumulated in their tissue. Specimens of cf. Haliclona (Haplosclerida) were sampled in the Maldivian archipelago. The aim was to optimize the method and to carry out a pilot study of the contamination of the related reef habitat. Particles were isolated, size fractioned, counted and submitted to morphological and chemical characterization. The constituting polymer was identified by infrared microspectroscopy for particles >25 µm, and by pyrolysis coupled with gas chromatography mass spectrometry for those <25 µm. Method recoveries were between 87 and 83% and limits of quantitation (LOQs) were between 6.6 and 30.2 ng/g. Analyses showed that 70% of the sponges presented plastic contamination, with an average of 1.2 particles/g tissue for the 25-150 µm size range, and a total plastic concentration of up to 4.8 µg/g in the 0.2-25 µm size range, with polyolefin being the most represented polymer in both size ranges. Overall, the study demonstrated the reliability of the proposed analytical workflow and of the use of sponges as biosamplers for plastic particles.