ALBERT

Description: Microbial co‐cultivation is employed for awakening silent biosynthetic gene clusters (BGCs) to enhance chemical diversity. However, the selection of appropriate partners for co‐cultivation remains a challenge. Furthermore, competitive interactions involving the suppression of BGCs or upregulation of known, functional metabolite(s) during co‐cultivation efforts is also common. Herein, we performed an alternative approach for targeted selection of the best co‐cultivation pair. Eight marine sediment‐derived fungi were classified as strong or weak, based on their anti‐phytopathogenic potency. The fungi were co‐cultured systematically and analyzed for their chemical profiles and anti-phytopathogenic activity. Based on enhanced bioactivity and a significantly different metabolite profile including the appearance of a co‐culture specific cluster, the co‐culture of Plenodomus influorescens (strong) and Pyrenochaeta nobilis (weak) was prioritized for chemical investigation. Large‐scale co‐cultivation resulted in isolation of five polyketide type compounds: two 12‐membered macrolides, dendrodolide E (1) and its new analog dendrodolide N (2), as well as two rare azaphilones spiciferinone (3) and its new analog 8a-hydroxy-spiciferinone (4). A well‐known bis‐naphtho‐γ‐pyrone type mycotoxin, cephalochromin (5), whose production was specifically enhanced in the co-culture, was also isolated. Chemical structures of compounds 1–5 were elucidated by NMR, HRMS and [] 20/D analyses. Compound 5 showed the strongest anti‐phytopathogenic activity against Xanthomonas campestris and Phytophthora infestans with IC50 values of 0.9 and 1.7 µg/mL, respectively.

Description: The fungal genus Pyrenochaetopsis is commonly found in soil, terrestrial, and marine environments, however, has received little attention as a source of bioactive secondary metabolites so far. In a recent work, we reported the isolation and characterization of three new anticancer decalinoyltetramic acid derivatives, pyrenosetins A-C, from the Baltic Fucus vesiculosus-derived endophytic fungus Pyrenochaetopsis sp. FVE-001. Herein we report a new pentacyclic decalinoylspirotetramic acid derivative, pyrenosetin D (1), along with two known decalin derivatives wakodecalines A (2) and B (3) from another endophytic strain Pyrenochaetopsis FVE-087 isolated from the same seaweed and showed anticancer activity in initial screenings. The chemical structures of the purified compounds were elucidated by comprehensive analysis of HR-ESIMS, FT-IR, [a]D, 1D and 2D NMR data coupled with DFT calculations of NMR parameters and optical rotation. Compounds 1-3 were evaluated for their anticancer and toxic potentials against the human malignant melanoma cell line (A-375) and the non-cancerous keratinocyte cell line (HaCaT). Pyrenosetin D (1) showed toxicity towards both A-375 and HaCaT cells with IC50 values of 77.5 and 39.3 μM, respectively, while 2 and 3 were inactive. This is the third chemical study performed on the fungal genus Pyrenochaetopsis and the first report of a pentacyclic decalin ring system from the fungal genus Pyrenochaetopsis.

Description: There is growing evidence that passive margin sediments in offshore settings host large volumes of fresh and brackish water of meteoric origin in submarine sub-surface reservoirs. Marine geophysical methods, in particular seismic reflection data, can help characterize offshore hydrogeological systems and yet the existing global database of industrial basin wide surveys remains untapped in this context. In this paper we highlight the importance of these data in groundwater exploration, by reviewing existing studies that apply physical stratigraphy and morpho-structural interpretation techniques to provide important information on—reservoir (aquifer) properties and architecture, permeability barriers, paleo-continental environments, sea-level changes and shift of coastal facies through time and conduits for water flow. We then evaluate the scientific and applied relevance of such methodology within a holistic workflow for offshore groundwater research.

Description: The study of wetlands is particularly important as these systems act as natural water purifiers and thus can act as sinks for contaminated particles. Wetland sediments are important as they provide an indication of potential contamination across temporal and spatial scales. The current study aimed to investigate the distributions of selected metals and nutrients in different sites in relation to sediment depth, and identify relationships among sediment metals. Significant differences in nutrient (i.e., N, P) and metal (i.e., K, Mg, Na, Fe, Cu, B) concentrations were found across study sites, whereas nutrients (i.e., N, P) and metals (i.e., Ca, Mg, Fe, Cu, Zn) were significantly different with sediment depths. When compared against Canadian sediment standards, most of the assessed metals were within the “no effect” level across the different sites and depths. The K, Ca, and Mg concentration showed extreme contamination across all sites and depths. The enrichment factor values for K, Ca, and Mg showed extremely high enrichment levels for all sites and sediment depths. The Na, Mn, Fe, Cu, Zn, and B concentration showed mostly background enrichment levels. All sediments across the different sites and sediment depths indicated deterioration of sediment quality. Pearson correlations suggest that most metals might have originated in a similar source as that of Mn and B, owing to a lack of significant differences. These results provide baseline information for the general management of the Nylsvley Wetland in relation to sediment metal pollution. The specific sources of metal contaminants also require further elucidation to further inform management efforts.

Description: Detailed multi-beam bathymetry, sub-bottom acoustic, and side-scan sonar observations of submerged canyons with tufa barriers were used to characterize the Zrmanja River karst estuary on the eastern Adriatic coast, Croatia. This unique karst environment consists of two submerged karst basins (Novigrad Sea and Karin Sea) that are connected with river canyons named Novsko Ždrilo and Karinsko Ždrilo. The combined use of high-resolution geophysical data with legacy topography and bathymetry data in a GIS environment allowed for the description and interpretation of this geomorphological setting in relation to the Holocene sea-level rise. The tufa barriers had a predominant influence on the Holocene flooding dynamics of the canyons and karst basins. Here, we describe the possible river pathways from the basins during the lowstand and the formation of a lengthening estuary during the Holocene sea-level rise. Based on the analyzed morphologies and the relative sea-level curve for the Adriatic Sea, the flooding of the Novsko Ždrilo occurred 9200 years before present (BP) and Karinsko Ždrilo was flooded after 8400 years BP. The combination of high-resolution geophysical methods gave an accurate representation of the karst estuarine seafloor and the flooding of semi-isolated basins due to sea-level rise.

Description: Many filamentous vibriophages encode virulence genes that lead to the emergence of pathogenic bacteria. Most genomes of filamentous vibriophages characterized up until today were isolated from human pathogens. Despite genome-based predictions that environmental Vibrios also contain filamentous phages that contribute to bacterial virulence, empirical evidence is scarce. This study aimed to characterize the bacteriophages of a marine pathogen, Vibrio alginolyticus (Kiel-alginolyticus ecotype) and to determine their role in bacterial virulence. To do so, we sequenced the phage-containing supernatant of eight different V. alginolyticus strains, characterized the phages therein and performed infection experiments on juvenile pipefish to assess their contribution to bacterial virulence. We were able to identify two actively replicating filamentous phages. Unique to this study was that all eight bacteria of the Kiel-alginolyticus ecotype have identical bacteriophages, supporting our previously established theory of a clonal expansion of the Kiel-alginolyticus ecotype. We further found that in one of the two filamentous phages, two phage-morphogenesis proteins (Zot and Ace) share high sequence similarity with putative toxins encoded on the Vibrio cholerae phage CTXΦ. The coverage of this filamentous phage correlated positively with virulence (measured in controlled infection experiments on the eukaryotic host), suggesting that this phage contributes to bacterial virulence.

Description: The migration of methane through the gas hydrate stability zone (GHSZ) in the marine subsurface is characterized by highly dynamic reactive transport processes coupled to thermodynamic phase transitions between solid gas hydrates, free methane gas, and dissolved methane in the aqueous phase. The marine subsurface is essentially a water-saturated porous medium where the thermodynamic instability of the hydrate phase can cause free gas pockets to appear and disappear locally, causing the model to degenerate. This poses serious convergence issues for the general-purpose nonlinear solvers (e.g., standard Newton), and often leads to extremely small time-step sizes. The convergence problem is particularly severe when the rate of hydrate phase change is much lower than the rate of gas dissolution. In order to overcome this numerical challenge, we have developed an all-at-once Newton scheme tailored to our gas hydrate model, which can handle rate-based hydrate phase change coupled with equilibrium gas dissolution in a mathematically consistent and robust manner.

Description: Latrunculia sponges represent a rich source of discorhabdin-type pyrroloiminoquinone alkaloids, a few of which comprise a dimeric structure. The anticancer-activity-guided isolation of the n-hexane subextract of the Antarctic deep-sea sponge Latrunculia biformis yielded the known compound (−)-(1R,2R,6R,8S,6'S)-discorhabdin B dimer (1) and two new derivatives, (−)-(1S,2R,6R,8S,6'S)-discorhabdin B dimer (2) and (−)-(1R,2R,6R,8S,6'S)-16',17'-dehydrodiscorhabdin B dimer (3). The chemical structures of compounds 1–3 were elucidated by means of HR-ESIMS, NMR, [], ECD spectroscopy, and a comparison with the previously reported discorhabdin analogs. Compounds 1 and 2 showed significant in vitro anticancer activity against the human colon cancer cell line (HCT-116), with IC50 values of 0.16 and 2.01 µM, respectively. Compared to monomeric discorhabdins, dimeric discorhabdins are very rare in Nature. This study adds two new discorhabdin dimers (2 and 3) to this small pyrroloiminoquinone subfamily. This is also the first report of compound 1 as a natural product and the first assessment of its in vitro anticancer activity.

Description: Sea foam harbors a diverse range of fungal spores with biological and ecological relevance in marine environments. Fungi are known as the producers of secondary metabolites that are used in health and agricultural sectors, however the potentials of sea foam-derived fungi have remained unexplored. In this study, organic extracts of six foam-derived fungal isolates belonging to the genera Penicillium, Cladosporium, Emericellopsis and Plectosphaerella were investigated for their antimicrobial activity against plant and human pathogens and anticancer activity. In parallel, an untargeted metabolomics study using UPLC-QToF–MS/MS-based molecular networking (MN) was performed to unlock their chemical inventory. Penicillium strains were identified as the most prolific producers of compounds with an average of 165 parent ions per strain. In total, 49 known mycotoxins and functional metabolites were annotated to specific and ubiquitous parent ions, revealing considerable chemical diversity. This allowed the identification of putative new derivatives, such as a new analog of the antimicrobial tetrapeptide, fungisporin. Regarding bioactivity, the Penicillium sp. isolate 31.68F1B showed a strong and broad-spectrum activity against seven plant and human pathogens, with the phytopathogen Magnaporthe oryzae and the human pathogen Candida albicans being the most susceptible (IC50 values 2.2 and 6.3 µg/mL, respectively). This is the first study mining the metabolome of the sea foam-derived fungi by MS/MS-based molecular networking, and assessing their biological activities against phytopathogens