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).

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.

Heavy metal and trace element concentrations in the blood of scalloped hammerhead sharks (Sphyrna lewini) from La Paz Bay, México.

Sharks are particularly susceptible to bioaccumulation due to their life history characteristics and trophic position within marine ecosystems. Despite this, studies of bioaccumulation cover only a small proportion of extant species. In this study we report concentrations of trace elements and heavy metals in blood samples of Sphyrna lewini for the first time. We report high concentrations of several trace elements and heavy metals, with concentrations of some elements exceeding the limit determined safe for human consumption. High elemental concentrations may reflect biochemical differences between blood plasma and other tissues; however, they may also be symptomatic of high levels of exposure triggered by anthropogenic activities. We also provide evidence of elemental accumulation through ontogeny, the nature of which differs from that previously reported. Ultimately, this baseline study increases our understanding of interspecific and intraspecific variation in bioaccumulation and ecotoxicology in elasmobranchs which may prove important in ensuring adequate management.

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.

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.

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.

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.

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.

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.

Diversity, host specificity and biogeography in the Cladocorynidae (Hydrozoa, Capitata), with description of a new genus.

The hydrozoan family Cladocorynidae inhabits tropical to temperate waters and comprises the two genera Pteroclava and Cladocoryne. Pteroclava lives in association with some octocorals and hydrozoans, whereas Cladocoryne is more generalist in terms of substrate choice. This work provides a thorough morpho-molecular reassessment of the Cladocorynidae by presenting the first well-supported phylogeny of the family based on the analyses of three mitochondrial and four nuclear markers. Notably, the two nominal genera were confirmed to be monophyletic and both morphological and genetic data led to the formal description of a new genus exclusively associated with octocorals, Pseudozanclea gen. nov. Maggioni & Montano. Accordingly, the diagnosis of the family was updated. The ancestral state reconstruction of selected characters revealed that the symbiosis with octocorals likely appeared in the most recent common ancestor of Pteroclava and Pseudozanclea. Additionally, the presence of euryteles aggregation in the polyp stage and the exumbrellar nematocyst pouches with euryteles represent synapomorphies of all cladocorynid taxa and probably emerged in their most recent common ancestor. The analysis of several Pteroclava krempfi colonies from Indo-Pacific and Caribbean localities associated with several host octocorals revealed a high intra-specific genetic variability. Single- and multi-locus species delimitations resulted in three to five species hypotheses, but the statistical analysis of morphometric data showed only limited distinction among the clades of P. krempfi. However, P. krempfi clades showed differences in both host specificity, mostly at the octocoral family level, and geographic distribution, with one clade found exclusively in the Caribbean Sea and the others found in the Indo-Pacific.

Ecological dependencies make remote reef fish communities most vulnerable to coral loss.

Ecosystems face both local hazards, such as over-exploitation, and global hazards, such as climate change. Since the impact of local hazards attenuates with distance from humans, local extinction risk should decrease with remoteness, making faraway areas safe havens for biodiversity. However, isolation and reduced anthropogenic disturbance may increase ecological specialization in remote communities, and hence their vulnerability to secondary effects of diversity loss propagating through networks of interacting species. We show this to be true for reef fish communities across the globe. An increase in fish-coral dependency with the distance of coral reefs from human settlements, paired with the far-reaching impacts of global hazards, increases the risk of fish species loss, counteracting the benefits of remoteness. Hotspots of fish risk from fish-coral dependency are distinct from those caused by direct human impacts, increasing the number of risk hotspots by ~30% globally. These findings might apply to other ecosystems on Earth and depict a world where no place, no matter how remote, is safe for biodiversity, calling for a reconsideration of global conservation priorities.

The mitochondrial genome of Nemalecium lighti (Hydrozoa, Leptothecata).

The hydrozoan species Nemalecium lighti (Hargitt, 1924) is widely distributed in tropical marine waters around the world. Here we report the complete linear mitochondrial genome of N. lighti from Sint Eustatius (Lesser Antilles). The mitochondrial genome with a length of 14,320 bp encodes for 13 protein-coding genes, two tRNA genes, and two rRNA genes. Gene arrangement differs from that found in other species of the same taxonomic order and a phylogenetic analysis shows that based on mitochondrial genes, N. lighti clusters outside of the Leptothecata, rendering the order paraphyletic.

Global tropical reef fish richness could decline by around half if corals are lost.

Reef fishes are a treasured part of marine biodiversity, and also provide needed protein for many millions of people. Although most reef fishes might survive projected increases in ocean temperatures, corals are less tolerant. A few fish species strictly depend on corals for food and shelter, suggesting that coral extinctions could lead to some secondary fish extinctions. However, secondary extinctions could extend far beyond those few coral-dependent species. Furthermore, it is yet unknown how such fish declines might vary around the world. Current coral mass mortalities led us to ask how fish communities would respond to coral loss within and across oceans. We mapped 6964 coral-reef-fish species and 119 coral genera, and then regressed reef-fish species richness against coral generic richness at the 1° scale (after controlling for biogeographic factors that drive species diversification). Consistent with small-scale studies, statistical extrapolations suggested that local fish richness across the globe would be around half its current value in a hypothetical world without coral, leading to more areas with low or intermediate fish species richness and fewer fish diversity hotspots.

Cryptic species and host specificity in the bryozoan-associated hydrozoan Zanclea divergens (Hydrozoa, Zancleidae).

Zanclea divergens is a tropical hydrozoan living in symbiotic association with bryozoans and currently reported from Papua New Guinea, Indonesia, and Maldives. Here, we used an integrative approach to assess the morpho-molecular diversity of the species across the Indo-Pacific. Phylogenetic and species delimitation analyses based on seven mitochondrial and nuclear loci revealed four well-supported molecular lineages corresponding to cryptic species, and representing a Pacific clade, an Indian clade, and two Red Sea clades. Since the general polyp morphology was almost identical in all samples, the nematocyst capsules were measured and analysed to search for possible fine-scale differences, and their statistical treatment revealed a significant difference in terms of length and width among the clades investigated. All Zanclea divergens specimens were specifically associated with cheilostome bryozoans belonging to the genus Celleporaria. The Pacific and Indian clades were associated with Celleporaria sp. and C. vermiformis, respectively, whereas both Red Sea lineages were associated with C. pigmentaria. Nevertheless, the sequencing of host bryozoans revealed that one of the Red Sea hydrozoan clades is associated with two morphologically undistinguishable, but genetically divergent, bryozoan species. Overall, our results show that Z. divergens is a species complex composed of morphologically cryptic lineages showing partially disjunct distributions and host specificity. The presence of two sympatric lineages living on the same host species reveal complex dynamics of diversification, and future research aimed at understanding their diversification process will likely improve our knowledge on the mechanisms of speciation among currently sympatric cryptic species.

Spatial variability of phthalates contamination in the reef-building corals Porites lutea, Pocillopora verrucosa and Pavona varians.

Microplastic pollution represents a serious hazard for the marine environment, including coral reefs. Scleractinian corals can easily mistake microplastics with their natural preys, and ingest them and all the annexed plasticizer additives. Here we selectively searched on field for five phthalates esters (PAEs) namely dibutyl-phthalate (DBP), benzylbutyl-phthalate (BBzP), diethyl-phthalate (DEP), Bis(2-ethylhexyl)-phthalate (DEHP), and dimethyl-phthalate (DMP) in the coral species Pocillopora verrucosa, Porites lutea and Pavona varians. Our data reveal that >95% of corals sampled were contaminated, with a maximum of 172.4 ng/g, a value 7 time-fold higher than those found in a previous study. The Σ5 PAEs showed an average of about 30 ng/g per coral, but no differences in PAEs contamination was detected between species, depth or reef exposure. Despite their effects on coral physiology are not yet known, PAEs should be now considered as a novel, and ubiquitous, form of contamination in corals.

Biocompatible solid-phase microextraction coupled to liquid chromatography triple quadrupole mass spectrometry analysis for the determination of phthalates in marine invertebrate.

This paper describes the development and validation of a new procedure for the determination of phthalates in marine invertebrates, based on biocompatible solid-phase microextraction (BioSPME) followed by liquid chromatography coupled to mass spectrometry. The importance of this application relies on the current use of marine organisms as bioindicators for microplastic contamination through the detection of phthalates in their tissues. Challenges originate from the availability and/or possible restriction on the use of the biological materials, the need for user- friendly procedures for simplifying and speeding up operations in the marine environments, and the ubiquitous presence of phthalates in the laboratory environments that may cause background contamination. BioSPME served as an effective solution to all these issues, owing to the extreme minimization of the sample manipulation. Sampling operations were limited to the direct transfer of small amounts of the biological materials (150 mg) inside glass vials capped with aluminium lids; extractions were carried out by ultrasonication in acetone followed by dilution in ultrapure water and BioSPME clean-up; Electrospray (ESI) LC-MS/MS was employed for the final analytical determinations on the purified extracts. Tests were carried out on samples belonging to three different phyla, namely Cnidaria, Porifera, and Mollusca obtained from Maldivian coral reef environments and used for a current microplastic contamination biomonitoring project. Overall, very good sensitivity and repeatability, with negligible back contamination of the blanks were observed. The limits of detections were between 0.2 and 2.1 ng/g and precision, calculated as relative standard deviation (RSD), was below 14% for all the tested phthalates.