BACKGROUND: Several studies have been reported previously on the bioactivities of different extracts of marine molluscs. Therefore, we decided to evaluate the cytotoxic and antimicrobial activities of S. pharaonis ink as a highly populated species in the Red Sea. We extracted the flavonoids from the ink and analyzed their composition. Then we evaluated systematically the cytotoxic and antimicrobial properties of this extract. A pharmacokinetic study was also conducted using SwissADME to assess the potential of the identified flavonoids and phenolic compounds from the ink extract to be orally active drug candidates.
RESULTS: Cytotoxic activity was evaluated against 5 cell lines (MCF7, Hep G2, A549, and Caco2) at different concentrations (0.4 µg/mL, 1.6 µg/mL, 6.3 µg/mL, 25 µg/mL, 100 µg/mL). The viability of examined cells was reduced by the extract in a concentration-dependent manner. The highest cytotoxic effect of the extract was recorded against A549 and Hep G2 cancer cell lines cells with IC50 = 2.873 and 7.1 µg/mL respectively. The mechanistic analysis by flow cytometry of this extract on cell cycle progression and apoptosis induction indicated that the extract arrests the cell cycle at the S phase in Hep G2 and MCF7, while in A549 cell arrest was recorded at G1 phase. However, it causes G1 and S phase arrest in Caco2 cancer cell line. Our data showed that the extract has significant antimicrobial activity against all tested human microbial pathogens. However, the best inhibitory effect was observed against Candida albicans ATCC 10,221 with a minimum inhibitory concentration (MIC) of 1.95 µg/mL. Pharmacokinetic analysis using SwissADME showed that most flavonoids and phenolics compounds have high drug similarity as they satisfy Lipinski\'s criteria and have WLOGP values below 5.88 and TPSA below 131.6 Å2.
CONCLUSIONS: S. pharaonis ink ethanolic extract showed a promising cytotoxic potency against various cell lines and a remarkable antimicrobial action against different pathogenic microbial strains. S. pharaonis ink is a novel source of important flavonoids that could be used in the future in different applications as a naturally safe and feasible alternative of synthetic drugs.

Microplastics are fast-emerging as another potential threat to already globally declining seagrass ecosystems, but there is a paucity of in situ surveys showing their accumulations. Here, we surveyed multiple Zostera marina L. meadows in 2020 and 2021 across Massachusetts, USA, for microplastic contamination, as well as identified factors related to patterns of accumulation. We found that microplastics were ubiquitous throughout all sites regardless of proximity to human development, with fibers being the most common microplastic type. In addition, we showed that accumulation of microplastics within seagrass meadows was related to epiphytic cover on leaves, plant morphology, and bulk-density in sediments. The results of this study provide the first in situ baseline microplastic concentrations on Z. marina plants and sediments for the temperate western North Atlantic. Additionally, we identify specific biotic and abiotic factors related to patterns of microplastic accumulation in these ecosystems.

Seagrass serves as a quintessential reservoir for obligate marine Lulworthiaceae fungi. Our current knowledge of the mycological diversity associated with seagrass in Hong Kong remains poor. We analyzed the diversity of fungi associated with the most widely distributed seagrass species in Hong Kong Halophila ovalis (Hydrocharitaceae), using a combination of culture-based methods and high-throughput amplicon sequencing. Halophilomyces hongkongensis, a novel fungal species in a newly proposed genus within the Lulworthiaceae family, was isolated from H. ovalis roots and rhizomes. The novel fungus showed distinct morphological characteristics, while both combined 18S-28S and internal transcribed spacer (ITS) phylogenetic trees based on maximum likelihood and Bayesian methods supported its discrimination from other existing Lulworthiaceae members. The ITS2 region in the Illumina sequencing results of multiple H. ovalis compartments, water, and adjacent non-seagrass sediments revealed continuous recruitment of H. hongkongensis by H. ovalis throughout the year despite dramatically fluctuating environmental conditions, with remarkably high proportions of this taxon found in root and rhizome internal tissues, possibly indicating a strong and specialized relationship established between the Lulworthiaceae fungal partner and its seagrass host. The inhibitory abilities exhibited by H. hongkongensis against Staphylococcus aureus SA29213 and ATCC 43300 (methicillin-resistant) may imply its capacity in producing (novel) antibacterial compounds. The discovery of H. hongkongensis as the first novel Lulworthiaceae taxon in Hong Kong, along with its distributional pattern in the seagrass meadow, provides valuable insights into the systematics and ecology of this strictly marine fungal family.

This study introduces a novel concept of \'Adaptively Stacked\' Species Distribution Models (AS-SDMs) to predict blue carbon habitat distribution, abundance, carbon stocks, and carbon sequestration potential in Orkney. AS-SDMs are built from Weighted Boosted Regression Trees (WBRTs) that adaptively stack blue carbon sediment thickness, sediment carbon content, and sequestration potential to predicted abundance. A novel method to describe substrate types by relative inputs of mud, sand, and gravel is detailed that better characterises the determining factors of seagrass, maerl, and horse mussel abundance. This study also introduces a novel use of indexes to mitigate double counting issues of mixed species distribution models. Seagrass, maerl, horse mussel, and mixed seagrass and maerl (SGM) habitats are estimated to cover a maximum area of 657 km2 in Orkney, have a total sediment carbon stock of 16 Mt. C, and sequester 6000 t C yr-1. Applying a conservative threshold of 50 % abundance to habitat predictions, six key potential areas of blue carbon offset projects are identified. These areas cover just over 9 km2, have a total carbon stock of 330,000 t C, and sequester 330 t C yr-1. When applied to UK carbon credit value, assuming integration with voluntary markets and compliance with accreditation criteria, the habitats in these areas have a potential value of £24.5 million. If applied as annual values, these areas have carbon stocks with a potential value of £0.93 million yr-1 and a carbon sequestration potential value of £24,000 yr-1.

Our study explored the lateral export of macroalgae and seagrass to the deep sea of the Northern South China Sea (NSCS). Particulate organic carbon (POC) collected from a depth of 500 m off southwestern Taiwan (station T) and Dongsha Atoll (station K) underwent environmental DNA (eDNA) and stable isotope assays. Metabarcoding using 18S V9 rDNA revealed lateral export of macrophyte detritus in NSCS. At station K, seagrass detritus predominated, while at station T, macroalgae-derived detritus was dominant. The consistency in the stable carbon isotope signature between POC and macrophytes indicates that stable carbon is an ideal bio-indicator for tracking macrophyte detritus destination and transformation after it has been laterally exported. Based on robust scientific methods, these findings provide valuable insights into the lateral export of macrophyte detritus to the deep sea in POC, influenced by habitat species, and shaped by distinct oceanographic physics around NSCS.

Pharmaceuticals like ibuprofen (IBU) entering marine environments are of great concern due to their increasing consumption and impact on wildlife. No information on IBU toxicity to seagrasses is yet available. Seagrasses form key habitats and are threatened worldwide by multiple stressors. Here, the responses of the seagrass Cymodocea nodosa to a short-term exposure (12 days) to environmentally realistic IBU concentrations (0.25-2.5-25 µg L-1), both at organism (plant growth) and sub-organism level (oxidative status, photosynthetic efficiency, and specialized metabolites production), were assessed in mesocosm. Chemical analyses to detect the presence of IBU and its metabolites in seawater and plants were also performed. IBU did not affect plant growth but caused physiological alterations which varied in severity depending on its concentration. Concentrations of 0.25 and 2.5 µg L-1 resulted in oxidative stress, but an increased antioxidant enzyme activity enabled plants to tolerate stress. A concentration of 25 µg L-1 caused greater oxidative stress, reduced antioxidant enzyme activity and specialized metabolites production, and impaired photosynthetic machinery functioning (particularly PSII). IBU was detected in seawater but not in plants suggesting no bioaccumulation. These findings indicate that C. nodosa could not withstand high IBU stress, and this could reduce its resilience to additional environmental stressors.

This study investigates the presence and impact of UV filters in Posidonia oceanica meadows in Formentera, a Mediterranean tourist hotspot. It highlights the distribution of inorganic (TiO2 and ZnO) and organic UV filters (UVFs) in different environmental matrices, their accumulation in seagrass tissues and their impact on the seagrass health. In the overlying and canopy waters of P. oceanica, Zn concentrations surpassed Ti, with three organic UVFs (benzophenone-3 [BP-3], avobenzone and homosalate [HMS]) consistently detected. Ti concentrations were generally higher than Zn in rhizosphere sediments, along with recurrent presence of octocrylene, HMS, 2-ethylhexyl methoxycinnamate (EHMC), and 4-methylbenzylidene camphor (4-MBC). Maximum Zn concentrations were found in canopy waters (3052.9 ng L-1). Both Ti and Zn were found in all P. oceanica tissues and leaf epiphytes across all study sites. Additional UVFs like octocrylene, avobenzone, and BP-8 were also detected in P. oceanica tissues and epiphytes. Elevated levels of octocrylene in leaf epiphytes (2112.1 ng g-1 dw) and avobenzone in leaves (364.2 ng g-1 dw) and leaf epiphytes (199.6 ng g-1 dw) were observed in the Port of La Savina, the island\'s main entry port. Octocrylene concentrations (up to 2575 ng g-1 dw) in rhizosphere sediments near sewage discharge points exceeded reported maxima, highlighting wastewater treatment plants as significant sources of organic UVFs. Correlational analyses suggested that the accumulation of octocrylene, avobenzone, and BP-3 negatively impacted P. oceanica\'s conservation status, affecting global density, density at 100 % cover, and leaf morphometry. Positive correlations were observed between leaf polyphenols (antioxidants) and concentrations of avobenzone, benzophenone-8 (BP-8), and BP-3, indicating potential oxidative stress induced by UVFs in P. oceanica. Our study underscores the pervasive presence of UV filters in P. oceanica habitats, with implications for seagrass health and conservation, especially in areas of high tourism and sewage discharge.

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Recent studies have shown that seagrasses could possess potential applications in the treatment of inflammatory disorders. Five seagrass species (Zostera muelleri, Halodule uninervis, Cymodocea rotundata, Syringodium isoetifolium, and Thalassia hemprichii) from the Great Barrier Reef (QLD, Australia) were thus collected, and their preliminary antioxidant and anti-inflammatory activities were evaluated. From the acetone extracts of five seagrass species subjected to 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging antioxidant assay, the extract of Z. muelleri had the highest activity (half minimal concentration of inhibition (IC50) = 138 µg/mL), with the aerial parts (IC50 = 119 µg/mL) possessing significantly higher antioxidant activity than the roots (IC50 ≥ 500 µg/mL). A human peripheral blood mononuclear cells (PBMCs) assay with bacterial lipopolysaccharide (LPS) activation and LEGENDplex cytokine analysis showed that the aerial extract of Z. muelleri significantly reduced the levels of inflammatory cytokines tumour necrosis factor alpha (TNF-α), interleukin (IL)-1β, and IL-6 by 29%, 74%, and 90%, respectively, relative to the LPS treatment group. The aerial extract was thus fractionated with methanol (MeOH) and hexane fraction, and purification of the MeOH fraction by HPLC led to the isolation of 4-hydroxybenzoic acid (1), luteolin (2), and apigenin (3) as its major constituents. These compounds have been previously shown to reduce levels of TNF-α, IL-1β, and IL-6 and represent some of the major bioactive components of Z. muelleri aerial parts. This investigation represents the first study of the antioxidant and anti-inflammatory properties of Z. muelleri and the first isolation of small molecules from this species. These results highlight the potential for using seagrasses in treating inflammation and the need for further investigation.

Genetic diversity can modulate a population\'s response to a changing environment and plays a critical role in its ecological function. While multiple processes act to maintain genetic diversity, sexual reproduction remains the primary driving force. Eelgrass (Zostera marina) is an important habitat-forming species found in temperate coastal ecosystems across the globe. Recent increases in sea surface temperatures have resulted in shifts to a mixed-annual life-history strategy (i.e., displaying characteristics of both annual and perennial meadows) at its southern edge-of-range. Given that mating systems are intimately linked to standing levels of genetic variation, understanding the scope of sexual reproduction can illuminate the processes that shape genetic diversity. To characterize edge-of-range eelgrass mating systems, developing seeds on flowering Z. marina shoots were genotyped from three meadows in Topsail, North Carolina. In all meadows, levels of multiple mating were high, with shoots pollinated by an average of eight sires (range: 3-16). The number of fertilized seeds (i.e., reproductive success) varied significantly across sires (range: 1-25) and was positively correlated with both individual heterozygosity and self-fertilization. Outcrossing rates were high (approx. 70%) and varied across spathes. No clones were detected, and kinship among sampled flowering shoots was low, supporting observed patterns of reproductive output. Given the role that genetic diversity plays in enhancing resistance to and resilience from ecological disturbance, disentangling the links between life history, sexual reproduction, and genetic variation will aid in informing the management and conservation of this key foundation species.