Click
here to close Hello! We notice that
you are using Internet Explorer, which is not supported by Echinobase
and may cause the site to display incorrectly. We suggest using a
current version of Chrome,
FireFox,
or Safari.
Mar Drugs
2020 Mar 31;184:. doi: 10.3390/md18040181.
Show Gene links
Show Anatomy links
Anti-Fouling Effects of Saponin-Containing Crude Extracts from Tropical Indo-Pacific Sea Cucumbers.
Kamyab E
,
Goebeler N
,
Kellermann MY
,
Rohde S
,
Reverter M
,
Striebel M
,
Schupp PJ
.
???displayArticle.abstract???
Sea cucumbers are bottom dwelling invertebrates, which are mostly found on subtropical and tropical sea grass beds, sandy reef flats, or reef slopes. Although constantly exposed to fouling communities in these habitats, many species are surprisingly free of invertebrate epibionts and microfouling algae such as diatoms. In our study, we investigated the anti-fouling (AF) activities of different crude extracts of tropical Indo-Pacific sea cucumber species against the fouling diatom Cylindrotheca closterium. Nine sea cucumber species from three genera (i.e., Holothuria, Bohadschia, Actinopyga) were selected and extracted to assess their AF activities. To verify whether the sea cucumber characteristic triterpene glycosides were responsible for the observed potent AF activities, we tested purified fractions enriched in saponins isolated from Bohadschia argus, representing one of the most active anti-fouling extracts. Saponins were quantified by vanillin-sulfuric acid colorimetric assays and identified by LC-MS and LC-MS/MS analyses. We were able to demonstrate that AF activities in sea cucumber extracts were species-specific, and growth inhibition as well as attachment of the diatom to surfaces is dependent on the saponin concentration (i.e., Actinopyga contained the highest quantities), as well as on the molecular composition and structure of the present saponins (i.e., Bivittoside D derivative was the most bioactive compound). In conclusion, the here performed AF assay represents a promising and fast method for selecting the most promising bioactive organism as well as for identifying novel compounds with potent AF activities for the discovery of potentially novel pharmacologically active natural products.
Figure 1. Structure of the saponin molecule “bivittoside D”, m/z 1449.687 [M + Na]+ [56], consisting of the glycone and aglycone moieties (produced with ChemDraw, version 16.0.1.4 (77)).
Figure 2. Phylogeny tree of the here studied sea cucumbers (CT = cuvierian tubule; adapted from [57,58,59,60].
Figure 3. Logarithmic response ratio (LRR) of C. closterium after exposure to three different concentrations (150, 15 and 1.5 µg mL−1) of nine sea cucumber extracts in total (genera Holothuria, Bohadschia and Actinopyga) for (A) suspended cells in the water and (B) attached to the surface of the incubation flask. Significant differences compared to the control (CT = control) are shown in Table S1.
Figure 4. Cluster dendrogram of sea cucumber species based on their studied saponin and sapogenin compositions (“average” distance type, log-transformed data, R version 1.2.5019).
Figure 5. Major saponin compounds detected in the studied sea cucumbers (peak area ≥ 104). (A) saponin diversity and relative intensity and (B) sapogenin (aglycon) diversity and relative intensity. Sample codes represent exact mass (M in Da), and retention time (T in min). Different colors represent the presence of sulphate groups (in blue), non-sulphate groups (in black) and pure compounds (in purple and red). Bubble size correlates with differences in relative peak areas of the respective molecules.
Figure 6. Absolute saponin concentration of the tested crude extracts. (a–c) indicate significant differences between different sea cucumber crude extracts. Kruskal–Wallis, Dunn’s method as a multiple comparison test. Significance level at p < 0.05 was applied.
Figure 7. Logarithmic response ratio (LRR) of C. closterium following exposure to B. argus extract fractions in suspended cells in the water (A) and attached to the substrate (B). Fr.1 and Fr.2: impure, Fr. 3: semi-pure, Fr. 4: pure singular saponin species (bivittoside D-like). a–d represent result of Kruskal-Wallis test; p < 0.05.
Figure 8. (A–C). The test organism C. closterium under the microscope (A), culture flasks demonstrating high growth rates (left, not-inhibited), medium growth rates (middle) and low growth rates (inhibited) of C. closterium (B), growth curve of C. closterium in 7 days in the stock solution (C).
Figure 9. Flow chart showing the applied procedure for isolating the bioactive saponin compounds (Cutignano et al., 2015; Ebada et al., 2008 [99,100]). Sample set 1 and 2 refers to the samples that were tested for anti-fouling (AF) activity in this study.
Affan,
OPTIMAL GROWTH CONDITIONS AND ANTIOXIDATIVE ACTIVITIES OF CYLINDROTHECA CLOSTERIUM (BACILLARIOPHYCEAE)(1).
2009, Pubmed
Affan,
OPTIMAL GROWTH CONDITIONS AND ANTIOXIDATIVE ACTIVITIES OF CYLINDROTHECA CLOSTERIUM (BACILLARIOPHYCEAE)(1).
2009,
Pubmed
Amara,
Antifouling processes and toxicity effects of antifouling paints on marine environment. A review.
2018,
Pubmed
Aslam,
Identifying metabolic pathways for production of extracellular polymeric substances by the diatom Fragilariopsis cylindrus inhabiting sea ice.
2018,
Pubmed
Bahrami,
Structure elucidation of five novel isomeric saponins from the viscera of the sea cucumber Holothuria lessoni.
2014,
Pubmed
,
Echinobase
Bahrami,
Discovery of novel saponins from the viscera of the sea cucumber Holothuria lessoni.
2014,
Pubmed
,
Echinobase
Bers,
Chemical defence in mussels: antifouling effect of crude extracts of the periostracum of the blue mussel Mytilus edulis.
2006,
Pubmed
Briand,
Marine antifouling laboratory bioassays: an overview of their diversity.
2009,
Pubmed
Cutignano,
Development and Application of a Novel SPE-Method for Bioassay-Guided Fractionation of Marine Extracts.
2015,
Pubmed
Dobretsov,
Mini-review: quorum sensing in the marine environment and its relationship to biofouling.
2009,
Pubmed
Dusane,
Anti-biofilm potential of a glycolipid surfactant produced by a tropical marine strain of Serratia marcescens.
2011,
Pubmed
Ebada,
Methods for isolation, purification and structural elucidation of bioactive secondary metabolites from marine invertebrates.
2008,
Pubmed
Elyakov,
A chemical investigation of the trepang (Stychopus japonicus Selenka): the structure of triterpenoid aglycones obtained from trepang glycosides.
1969,
Pubmed
Faulkner,
Marine natural products.
1994,
Pubmed
Fusetani,
Biofouling and antifouling.
2004,
Pubmed
,
Echinobase
Gerhard,
Phytoplankton community responses to temperature fluctuations under different nutrient concentrations and stoichiometry.
2019,
Pubmed
Grauso,
Molecular Networking-Based Analysis of Cytotoxic Saponins from Sea Cucumber Holothuria atra.
2019,
Pubmed
,
Echinobase
Haug,
Antibacterial activity in Strongylocentrotus droebachiensis (Echinoidea), Cucumaria frondosa (Holothuroidea), and Asterias rubens (Asteroidea).
2002,
Pubmed
,
Echinobase
Hellio,
Inhibition of marine bacteria by extracts of macroalgae: potential use for environmentally friendly antifouling paints.
2001,
Pubmed
,
Echinobase
Hiai,
Color reaction of some sapogenins and saponins with vanillin and sulfuric acid.
1976,
Pubmed
Honey-Escandón,
Biological and taxonomic perspective of triterpenoid glycosides of sea cucumbers of the family Holothuriidae (Echinodermata, Holothuroidea).
2015,
Pubmed
,
Echinobase
Ioannou,
9,11-Secosterols with antiproliferative activity from the gorgonian Eunicella cavolini.
2009,
Pubmed
Iorizzi,
Chemical and biological investigation of the polar constituents of the starfish Luidia clathrata, collected in the Gulf of Mexico.
1995,
Pubmed
,
Echinobase
Kalinin,
Triterpene glycosides of sea cucumbers (Holothuroidea, Echinodermata) as taxonomic markers.
2015,
Pubmed
,
Echinobase
Kitagawa,
Marine natural products. XIV. Structures of echinosides A and B, antifungal lanostane-oligosides from the sea cucumber Actinopyga echinites (Jaeger).
1985,
Pubmed
,
Echinobase
Kubanek,
Multiple defensive roles for triterpene glycosides from two Caribbean sponges.
2002,
Pubmed
Lorent,
Induction of highly curved structures in relation to membrane permeabilization and budding by the triterpenoid saponins, α- and δ-Hederin.
2013,
Pubmed
Lorent,
The amphiphilic nature of saponins and their effects on artificial and biological membranes and potential consequences for red blood and cancer cells.
2014,
Pubmed
Majik,
Next generation biofilm inhibitors for Pseudomonas aeruginosa: Synthesis and rational design approaches.
2014,
Pubmed
McKenzie,
The echinoderm surface and its role in preventing microfouling.
1996,
Pubmed
,
Echinobase
Mellado,
Steroids from the Antarctic octocoral Anthomastus bathyproctus.
2005,
Pubmed
Miller,
Molecular phylogeny of extant Holothuroidea (Echinodermata).
2017,
Pubmed
,
Echinobase
Ortlepp,
Antifouling activity of bromotyrosine-derived sponge metabolites and synthetic analogues.
2007,
Pubmed
Qi,
Antifouling Compounds from Marine Invertebrates.
2017,
Pubmed
Qi,
Antifouling and antibacterial compounds from the gorgonians Subergorgia suberosa and Scripearia gracillis.
2008,
Pubmed
Qian,
Natural products as antifouling compounds: recent progress and future perspectives.
2010,
Pubmed
Salta,
Designing biomimetic antifouling surfaces.
2010,
Pubmed
Singh,
Potential applications of microbial surfactants in biomedical sciences.
2004,
Pubmed
Stebbing,
Hormesis--the stimulation of growth by low levels of inhibitors.
1982,
Pubmed
Stebbing,
A theory for growth hormesis.
1998,
Pubmed
Thrane,
Spectrophotometric Analysis of Pigments: A Critical Assessment of a High-Throughput Method for Analysis of Algal Pigment Mixtures by Spectral Deconvolution.
2015,
Pubmed
Van Dyck,
Qualitative and quantitative saponin contents in five sea cucumbers from the Indian ocean.
2010,
Pubmed
,
Echinobase
Van Dyck,
Elucidation of molecular diversity and body distribution of saponins in the sea cucumber Holothuria forskali (Echinodermata) by mass spectrometry.
2009,
Pubmed
,
Echinobase
Van Dyck,
Localization of secondary metabolites in marine invertebrates: contribution of MALDI MSI for the study of saponins in Cuvierian tubules of H. forskali.
2010,
Pubmed
,
Echinobase
Van Dyck,
The triterpene glycosides of Holothuria forskali: usefulness and efficiency as a chemical defense mechanism against predatory fish.
2011,
Pubmed
,
Echinobase
Wimpenny,
Heterogeneity in biofilms.
2000,
Pubmed
Wright,
Anti-malarial, anti-algal, anti-tubercular, anti-bacterial, anti-photosynthetic, and anti-fouling activity of diterpene and diterpene isonitriles from the tropical marine sponge Cymbastela hooperi.
2011,
Pubmed
