Antimicrobial activities of extracts from Indo-Pacific marine plants against marine pathogens and saprophytes

This study is the second of two surveys designed to systematically screen extracts from marine plants for antimicrobial effects against ecologically relevant marine microorganisms, and to compare results on a geographical basis. In the preceding survey, extracts from tropical Atlantic marine algae and seagrasses were screened in growth inhibition assays against the pathogenic fungus Lindra thalassiae, the saprophytic fungus Dendryphiella salina, the saprophytic stramenopiles, Halophytophthora spinosa and Schizochytrium aggregatum, and the pathogenic bacterium Pseudoaltermonas bacteriolytica. In this study, the same assay microorganisms were used to examine the antimicrobial effects of lipophilic and hydrophilic extracts from 54 species of marine algae and two species of seagrasses collected from Indo-Pacific reef habitats. Overall, 95% of all species surveyed in this study yielded extracts that were active against one or more, and 77% yielded extracts that were active against two or more assay microorganisms. Broad-spectrum activity against three or four assay microbes was observed in the extracts from 50 to 21% of all species, respectively. Extracts from the green alga Bryopsis pennata and the red alga Portieria hornemannii inhibited the growth of all assay microorganisms. Given that antimicrobial activity was prevalent among extracts of Indo-Pacific marine plants, it is interesting to note that the inhibitory effects of each extract varied considerably between the assay microorganisms. Overall, H. spinosa and D. salina were the most susceptible while L. thalassiae, S. aggregatum, and P. bacteriolytica were the most resistant to the extracts tested. These results provide good evidence that antimicrobial chemical defenses are widespread among Indo-Pacific marine plants. Further, the activity profiles of plant extracts suggest that antimicrobial secondary metabolites can have pathogen-selective or broad-spectrum effects. To confirm these results, chemical studies will be needed to isolate and characterize the compounds responsible for the observed antimicrobial activities.     

Antimicrobial activity of Red Sea corals

Scleractinian corals and alcyonacean soft corals are the two most dominant groups of benthic marine organisms inhabiting the coral reefs of the Gulf of Eilat, northern Red Sea. Antimicrobial assays performed with extracts of six dominant Red Sea stony corals and six dominant soft corals against marine bacteria isolated from the seawater surrounding the corals revealed considerable variability in antimicrobial activity. The results demonstrated that, while the majority (83%) of Red Sea alcyonacean soft corals exhibited appreciable antimicrobial activity against marine bacteria isolated from the seawater surrounding the corals, the stony corals had little or no antimicrobial activity. From the active soft coral species examined, Xenia macrospiculata exhibited the highest and most potent antimicrobial activity. Bioassay-directed fractionation indicated that the antimicrobial activity was due to the presence of a range of compounds of different polarities. One of these antibiotic compounds was isolated and identified as desoxyhavannahine, with a minimum inhibitory concentration (MIC) of 48 μg ml⁻¹ against a marine bacterium. The results of the current study suggest that soft and hard corals have developed different means to combat potential microbial infections. Alcyonacean soft corals use chemical defense through the production of antibiotic compounds to combat microbial attack, whereas stony corals seem to rely on other means.     

Interspecific differences in fouling of two congeneric ascidians (Eudistoma olivaceum and E. capsulatum): is surface acidity an effective defense?

Ascidian specimens were collected from the Indian River Lagoon, Florida, USA, in November 1987. Eudistoma olivaceum (Van Name) was fouled by only two species of epifaunal invertebrates and the percent cover of epibionts was negligible. Colonies of E. capsulatum (Van Name) were fouled by up to 17 species of epifaunal invertebrates, epibionts covered an average of 12.5% of the surface area of these colonies. Extracts of E. olivaceum exhibited potent cytotoxic, antimicrobial and antiviral activity in laboratory bioassays, while the activity of extracts of E. capsulatum was insignificant. The toxicity of extracts of each ascidian was negatively related to the percent cover of fouling organisms. Surface acidity of each ascidian was positively related to the percent cover of epibionts: E. olivaceum was nearly neutral (pH=6), while E. capsulatum was highly acidic (pH=1 to 2). Larval settlement and growth of ancestrulae of the cheilostome bryozoan Bugula neritina (Linnaeus) were inhibited at very low concentrations of crude extract of E. olivaceum. Crude extracts of E. capsulatum inhibited settlement and growth of B. neritina only at concentrations of approximately an order of magnitude greater than those of E. olivaceum. Differences in the degree to which these congeneric ascidians are fouled is related to differences in the chemical defenses they possess. Acidity is not an effective inhibitor of settlement by epifaunal invertebrates.     

Antimicrobial activities of extracts from tropical Atlantic marine plants against marine pathogens and saprophytes

Studies investigating disease resistance in marine plants have indicated that secondary metabolites may have important defensive functions against harmful marine microorganisms. The goal of this study was to systematically screen extracts from marine plants for antimicrobial effects against marine pathogens and saprophytes. Lipophilic and hydrophilic extracts from species of 49 marine algae and 3 seagrasses collected in the tropical Atlantic were screened for antimicrobial activity against five ecologically relevant marine microorganisms from three separate kingdoms. These assay microbes consisted of the pathogenic fungus Lindra thalassiae, the saprophytic fungus Dendryphiella salina, the saprophytic stramenopiles, Halophytophthora spinosa and Schizochytrium aggregatum, and the pathogenic bacterium Pseudoaltermonas bacteriolytica. Overall, 90% of all species surveyed yielded extracts that were active against one or more, and 77% yielded extracts that were active against two or more assay microorganisms. Broad-spectrum activity against three or four assay microorganisms was observed in the extracts from 48 and 27% of all species, respectively. The green algae Halimeda copiosa and Penicillus capitatus (Chlorophyta) were the only species to yield extracts active against all assay microorganisms. Among all assay microorganisms, both fungi were the most resistant to the extracts tested, with less than 21% of all extracts inhibiting the growth of either L. thalassiae or D. salina. In contrast, over half of all lipophylic extracts were active against the stramenopiles H. spinosa and S. aggregatum, and the bacterium P. bacteriolytica. Growth sensitivity to hydrophilic extracts varied considerably between individual assay microorganisms. While 48% of all hydrophilic extracts were active against H. spinosa, 27% were active against P. bacteriolytica, and only 14% were active against S. aggregatum. Overall, more lipophilic extracts inhibited microbial growth than hydrophilic extracts. The variability observed in the antimicrobial effects of individual extracts against each assay microorganism reflects the importance of choosing appropriate test microbes in assays from which ecologically relevant information is sought. Results from this survey demonstrate that antimicrobial activities are prevalent among extracts from marine algae and seagrasses, suggesting that antimicrobial chemical defenses are widespread among marine plants.     

Antimicrobial activity of extracts of Caribbean gorgonian corals

Extracts of 39 species of Caribbean gorgonians were tested for antimicrobial activity against 15 strains of marine bacteria. The bacteria consisted of three opportunistic pathogens, Vibrio parahaemolyticus, Leucothrix mucor, and Aerococcus viridans, and 12 strains isolated from either healthy or decayed gorgonians. Overall, only 15% (79 out of 544) of the tests resulted in antibacterial activity with 33% (13 out of 39) of the gorgonians inhibiting only one bacterial strain and 23% (9 out of 39) showing no activity. The extracts of four Pseudopterogorgia species showed relatively high levels of activity, inhibiting 43 to 86% of the bacterial strains. The potency of the active Pseudopterogorgia species was variable, however, and three additional Pseudopterogorgia species were inactive against all bacterial strains. With the exception of one sensitive strain, Vibrio species were resistant to gorgonian metabolites. Our results indicate that organic extracts of most Caribbean gorgonians do not possess potent, broad-spectrum antibacterial activity inhibitory to the growth of opportunistic marine pathogens and bacteria associated with healthy and decayed gorgonian surfaces. These findings suggest that the inhibition of bacterial growth is not the primary ecological function of gorgonian secondary metabolites and that bacteria may not be important selective agents in the evolution of gorgonian secondary chemistry.     

Ecological roles of natural products from the marine sponge Geodia corticostylifera

In the Brazilian coast, high numbers of the small brittle star Ophiactis savignyi usually live associated with the sponge Geodia corticostylifera (Demospongiae, Geodidae), but not with other sympatric sponge species. In order to check whether this association was related only with the physical shelter provided by the sponge body or was chemically mediated, the crude organic extract of G. corticostylifera was added to sponge mimics made of phytagel and spongin skeleton. Control and treated mimics were simultaneously offered to previously sponge-associated O. savignyi in both static seawater and flow-through laboratory experiments. Ophiuroids were allowed to move towards the preferred mimic. The defensive properties of the sponge extract against fish predation and fouling were also evaluated. Chemotaxis assays showed that symbiotic ophiuroids were able to chemically recognize its host sponge, moving significantly more towards mimics containing G. corticostylifera extract. Chemical deterrence assays showed that the natural concentration of the extract of this sponge was also able to inhibit generalist fish predation on field experiments and the attachment of the common mussel Perna perna in laboratory assays. These results indicate that the crude extract of G. corticostylifera plays multiple functions in the marine environment, presumably being responsible for a closer association of this sponge with O. savignyi, providing protection for this ophiuroid and inhibition of epibionts on itself.     

Antibacterial and antimutagenic activitiy of extracts and essential oil from (Tunisian) Pistacia lentiscus

Aqueous and flavonoid-enriched extract as well as essential oil (EO) obtained from leaves of Pistacia lentiscus were assessed for antibacterial and antimutagenic activities. Antibacterial activity of different extracts and EO were evaluated against six bacterial strains. A marked inhibitory effect was observed against Salmonella typhimurium, whereas lower activity was observed against Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella enteritidis. EO showed significant inhibitory effects against Salmonella typhimurium, Salmonella enteritidis and Staphylococcus aureus. The antimutagenic activity of the different extracts against Aflatoxin B₁ (AFB₁) and sodium azide was demonstrated with the Salmonella typhymurium assay. The number of revertants per plate decreased significantly when the plant extracts were added to the assay system using Salmonella typhimurium TA100, TA98 and TA1535.     

Callyspongia samarensis (Porifera) extracts exhibit anticancer activity and induce bleaching in Porites cylindrica (Scleractinia)

Marine sponges can produce allelopathic compounds with specific roles in the competition for benthic space. Here we demonstrate that extracts from Callyspongia samarensis (Phylum Porifera) accelerate bleaching in Porites cylindrica (Phylum Cnidaria, Order Scleractinia) and exhibit in vitro anticancer activity. A column chromatography fraction and HPLC fraction, purified from the crude methanol extract of C. samarensis, were incorporated into agar gel cubes at natural concentrations and tested on P. cylindrica corals in a laboratory assay. Statistical analysis of percent bleached area and maximum quantum yield showed that a significant difference existed between P. cylindrica nubbins that were exposed to C. samarensis extracts versus the control group. This suggests that C. samarensis contains allelopathic compounds that can cause bleaching in P. cylindrica, possibly leading to tissue necrosis and death. Furthermore, the aforementioned HPLC fraction exhibited significant growth inhibition of the HCT-116 (human colon cancer) cell line. Understanding the strategies by which sponges exert dominance over other organisms is important because it provides information about the ecological roles of sponge allelopathy and can result in the discovery of compounds with biomedical potential.     

Turning the game around: toxicity in a nudibranch-sponge predator–prey association

Escalation theory proposes enemy-related selection as the most relevant factor of natural selection among individual organisms. When hazardous to predators, prey might be considered enemies that influence predator evolution. Opisthobranch molluscs that prey on chemically defended prey are an interesting study case on this subject. Predation on chemically defended species paved the way for opisthobranchs to enter in an arms race, developing means to detoxify and/or excrete harmful compounds, which led to the sequestration of those compounds and their self-defensive use, an escalation of defenses. Here we aim to understand whether the opisthobranch predator is better protected than its chemically defended prey, using as predator–prey model, a nudibranch (Hypselodoris cantabrica) and the sponge it preys upon (Dysidea fragilis), and from which it obtains deterrent chemical compounds. Specimens of both species were collected on the Portuguese coast, and their crude extracts were analyzed and used in palatability tests. Nudibranchs revealed a higher natural concentration of crude extract, probably due to a progressive accumulation of the compounds. Both predator and prey extracts revealed similar mixtures of deterrent metabolites (furanosesquiterpenes). Palatability tests revealed a more effective deterrence in the nudibranch extracts because significant rejection rates were observed at lower concentrations than those necessary for the sponge extracts to have the same effect. We concluded that the predator is chemically better protected than its prey, which suggests that its acquisition of chemical defenses reveals a defensive escalation.     

Enzymes from marine phycophages that degrade cell walls of seaweeds

Agarase, cellulase and alginate lyase activities from crude extracts of Aplysia dactylomela Rang, Haliotis coccinea canariensis Nordsieck, Littorina striata King et Broderip and Diadema antillarum Phillipi were measured in vitro to compare digestive efficiencies against several components of complex seaweed cell walls. Commercial abalone acetone powder (AAP, an extract from Haliotis sp.; Sigma, Ref. A-7514) and purified (Sigma, Ref. A-6306) and non-purified agarases from Pseudomonas atlantica were used with the same objective. Optimum conditions for agarase and cellulase activities were 40°C and pH 6.0. For alginate lyase, optimum temperature and pH were species-dependent. Highest reducing sugar release was shown by crude extracts from A. dactylomela. These crude extracts displayed high agarase activity compared with bacterial agarases at 40°C, and were significantly higher at 25°C. AAP and crude extracts from L. striata and D. antillarum exhibited high specific activities on all the substrates. Cold extract from Gracilaria spp. was the best substrate with which to measure agarase activity.     

Toxicity and palatability of some British ascidians

Bioassays were conducted to test crude extracts of nine temperate-water ascidians for bioactivity against a range of marine organisms. The extracts generally produced moderate to weak effects, with the exception of those from the ascidian Clavelina lepadiformis, which consistently produced high mortalities against invertebrates. Some of these were then tested in simple palatability assays for feeding-deterrent effects. In general, no strong effects were detected for assays with shore crabs (Carcinus maenas) and fishes (the goby Pomatoschistus minutus and the grey mullet Chelon labrosus). Feeding-deterrent effects were observed with Carcinus maenas for the extracts of Aplidium proliferum, Botryllus schlosseri and Dendrodoa grossularia. It is postulated that these ascidians are unpalatable, but no toxic effects are evident.     

Parrotfish predation on cryptic sponges of Caribbean coral reefs

Some sponge species that live in crevices in the reef frame appear to be restricted to their cryptic habitat by predation. When cryptic sponges were excavated, on Guigalotupo reef, San Blas, Panama, exposing them to potential predators, they were eaten by fishes that are generally considered to be herbivores, primarily parrotfishes of the genus Sparisoma: S.aurofrenatum (Cuvier & Valenciennes), S.viride (Bonnaterre), and S.chrysopterum (Bloch & Schneider). Of the 9150 bites observed to be taken by these species during paired (i.e., with sponges versus without sponges) trials conducted in defined feeding areas during 1986, 1987, and 1988, 72% (i.e. 6581 bites) were on cryptic sponges, even though these were only offered during half of the total observation time and never constituted more than 7% of the cover of the feeding observation areas. Individual parrotfish returned over and over to take bites of the exposed cryptic sponges until they were entirely consumed. They vigorously chased each other away from the sponges, but exhibited no such defense of their usual algal foods. A total of 18 sponge species were tested. Of the cryptic and semi-cryptic sponge species tested, only one of six was rejected by the parrotfish. Two of these six sponge species were consistently consumed entirely, and two were consumed entirely whenever their surfaces were sliced off with a razor blade, demonstrating that these sponges concentrate defenses against predators in their surfaces. One semi-cryptic species and one semi-exposed species were fed upon, but not entirely consumed. By contrast, 11 of 12 of the exposed and semi-exposed species were rejected. Cryptic sponges grew out of their cavities in the reef only when protected by seasonally thick mats of macroalgae or by cages that excluded fish. Received: 10 January 1997 / Accepted: 4 February 1997     

Oviposition stimulants and deterrents control acceptance ofAlliaria petiolata byPieris rapae andP. napi oleracea

Summary Differential acceptance of garlic mustard,Alliaria petiolata byPieris rapae L. andP. napi oleracea is explained by their differential sensitivities to oviposition stimulants and deterrents in the plant. Fractions containing the stimulants and deterrents were isolated by solvent partitioning between water and n-butanol and by open-column chromatography followed by HPLC.P. napi oleracea showed no preference when offered a choice ofA. petiolata or cabbage, but was strongly stimulated to oviposit by post-butanol water extracts ofA. petiolata. The most abundant glucosinolate in this extract was identified as sinigrin, which could explain the high degree of stimulatory activity.P. rapae preferred cabbage plants overA. petiolata, and the relatively low stimulatory activity was also associated with the glucosinolate-containing aqueous extract. However, this species was strongly stimulated by a fraction that contained small amounts of glucotropaeolin along with unknown compounds. Deterrents to both species were found in the butanol extract fromA. petiolata, andP. napi oleracea was more sensitive thanP. rapae to these deterrents. Some HPLC fractions from the BuOH extract were strongly deterrent toP. napi oleracea, but were inactive toP. rapae. The ecological significance of these behavioral differences between the twoPieris species is discussed.     

Impact of conditions of cultivation and adsorption on antimicrobial activity of marine bacteria

The potency of free-living and animal-associated marine bacteria to produce antimicrobial substances has been studied in 491 strains isolated from northern and southern parts of the Pacific Ocean. A total of 26% (126 out of 491) of the strains examined produced antimicrobial compounds against 11 test bacterial strains (TBS) including the fish pathogens Aeromonas hydrophila and Vibrio anquillarum. Antimicrobial substances (AS) produced by marine bacteria were especially active against Staphylococcus epidermidis, Proteus vulgaris, Enterococcus faecalis, and Candida albicans. Twelve strains, isolated from different sources, were chosen as promising candidates, producing a number of AS. Production of AS varied within 24 to 72 h, increasing in a culture medium based on natural sea water with Br-ions, and after attachment to polymeric surfaces. In order to study the influence of adsorption, selected strains with a high potential for antimicrobial production were cultivated on polymeric surfaces with different hydrophobicities and chemical functionalities. These parameters of the surface hydrophobicity (measured by means of water contact angles) and chemical functionality of the surfaces were manipulated using the photo- and thermochemistry of a polymeric system (diazo-naphto-quinone/novolak) commonly used as a photoresistant material in semiconducto-manufacturing. The highest antimicrobial activities occurred on hydrophilic surfaces (standard exposed photoresistant films), whereas the number of attached cells was greater on hydrophobic surfaces, characterized as unexposed resistant films. These results suggest that the chemical nature of induced hydrophilicity may also be a major factor in controlling antimicrobial activity of adsorbed bacteria. Received: 5 March 1997 / Accepted: 24 August 1997     

Larvicidal potential of different solvent extracts and oleanolic acid derivative from Coccinia indica against vector mosquitoes

The mosquito larvicidal activities of different solvent extracts (chloroform, ethyl acetate, and methanol) from the leaves of Coccinia indica were studied. Among the extracts tested, the methanol extract showed the highest larvicidal activity against early fourth instar larvae of Culex quinquefasciatus (LC₅₀?=?142?mg?L^?1), Aedes aegypti (LC₅₀?=?158?mg?L^?1) and Anopheles stephensi (LC₅₀?=?161?mg?L^?1) after 24?h of exposure period. The lowest larval mortality was observed with the ethyl acetate extract with the LC₅₀ values of 927, 872, and 730?mg?L^?1 against Cx. quinquefasciatus, Ae. aegypti, and An. stephensi, respectively. The chloroform extract showed the moderate larvicidal activity against the mosquito species tested with the LC₅₀ values from 446 to 546?mg?L^?1. A new triterpene was isolated from the methanol extract using bio-assay guided separation and identified as an oleanolic acid derivative by NMR, IR and mass spectral studies. The compound showed prominent larvicidal activity against Cx. quinquefasciatus, Ae. aegypti and An. stephensi with the LC₅₀ values of 5.6, 5.0, and 4.8?mg?L^?1, respectively. The results of the present investigation offer an opportunity for developing alternatives to rather expensive and environmentally hazardous organic insecticides.     

Protective effect of Kappaphycus alvarezii (Rhodophyta) extract against DNA damage induced by mercury chloride in marine fish

Marine organisms are continuously exposed to agents, both exogenous and endogenous, that damage DNA. Consequently, it is important to determine the ability of compounds to provide protection against damaging chemicals. The aim of this study was to evaluate the anti-genotoxic activity of crude aqueous extracts of Kappaphycus alvarezii (Rhodophyceae), collected from the Southeast coast of India. This study focused on possible anti-genotoxic potential of aqueous extract of K. alverazii to interfere with clastogenicity induced by mercury chloride (HgCl₂) in marine fish, Therapon jarbua as measured by cytogenetic endpoints such as cell viability and comet assay. In the first set of experiments, fish were exposed to a single treatment of Hg at 0.125, 0.25, 0.5, 1, or 2?ppm along with controls. Mercury exposure produced significant DNA damage in all comet classes, maximum as >79% (Class 4) at 0.5, 1, and 2?ppm exposure in a time dependent manner. Algal extract did not induce genotoxicity when given alone and prevented Hg-induced genotoxicity. The algal extract reduction in genotoxicity was significant but not time- and concentration-dependent. Results suggested that under present experimental conditions, K. alvarezii extract exhibit potent anti-genotoxicity effects in this fish model; and thus these extracts may be recommended as a supplement in fish meal and may benefit humans ingesting Hg-contaminated fish.     

Production of an antimicrobial cytochalasan by an endophytic Chaetomium globosum HYML55 from Hypericum mysorense and its RNA secondary structure analysis

The mutualistic benefits and disadvantages of non-clavicipitaceous endophytes for their hosts are not well studied and are postulated as being distinct from free-living or pathogenic counterparts. The defensive metabolites and antioxidants produced by endophytes are postulated to be the benefits offered for the hosts. Therefore, the antimicrobial and free-radical-scavenging activities of a foliar endophyte Chaetomium globosum HYML55 isolated from Hypericum mysorense of Western Ghats, India were studied. RNA secondary structure analysis refuted the unique biotope postulation because strain HYML55 was more similar to a coprophilous strain than to endophytic strains. Ethyl acetate extract of the fungus inhibited bacteria and fungi. The extract also quenched 98.65% 1,1-diphenyl- 2-picrylhydrazyl radicals with total antioxidant capacity of 13.62 mg ascorbic acid equivalent/g. A yellow amorphous antimicrobial compound purified from the extract was identified as chaetoglobosin F, which had a minimum inhibitory concentration of 7.8–15.6 μg/mL, except against Pseudomonas aeruginosa. This is the first report on the antimicrobial activity of chaetoglobosin F and other bioactivities of any endophytes from H. mysorense. The study emphasises the inter-habitat cycling of endophytes and possible benefits of C. globosum for the host. Chaetoglobosin F is an anti-inflammatory and anti-neoplastic compound, and C. globosum HYML55 could be exploited for the industrial production of this compound.     

A comprehensive evaluation of the potential chemical defenses of antarctic ascidians against sympatric fouling microorganisms

The present study analyzed the bioactivity of whole body extracts from six solitary and eight colonial ascidian taxa against 20 sympatric bacterial isolates and one sympatric diatom species from the Western Antarctic Peninsula. Ascidians had crude lipophilic and hydrophilic extracts assayed against 20 bacterial strains. The lipophilic extract of one ascidian caused growth inhibition in all bacterial isolates at 3× tissue-level concentrations. The lipophilic and hydrophilic extracts were fractionated into seawater-soluble and insoluble fractions and assayed at three concentrations against a sympatric diatom species. Significant diatom mortality was detected at 3× and 1× concentrations in all but one ascidian taxon. Lipophilic fractions caused higher diatom mortality than hydrophilic extracts. The specificity of secondary metabolites against diatom fouling and the lack of activity against bacteria suggest high selective pressure for chemical defenses against diatom fouling or the potential that bacterial pathogens are controlled by the ascidian immune system.     

Evaluation of the antimutagenic and antiradical potentials of extracts from the tubers of (Tunisian) Cyperus rotundus

The mutagenic potential of aqueous, total oligomers flavonoids (TOF), ethyl acetate and methanol extracts from tubers of Cyperus rotundus L. was assessed using Ames Salmonella tester strains TA98 and TA100, and SOS chromotest strain Escherichia coli PQ37 with and without metabolic activation (S9). None of the different extracts produced a mutagenic effect. Likewise, the antimutagenicity of the same extracts was tested using the “Ames test” and the “SOS chromotest”. Our results showed that C. rotundus extracts possess antimutagenic effects against S. typhimurium TA98 and TA100 strains towards the mutagen aflatoxin B1 (AFB1), similar to E. coli PQ37 strain against AFB1 and Nifuroxazide mutagens using the SOS chromotest assay. A free radical scavenging test was used in order to explore the antioxidant capacity of the extracts obtained from the tubers of C. rotundus. TOF, ethyl acetate and methanol extracts showed an important free radical scavenging activity towards the 1,1-diphenyl-2-picrylhydrazine (DPPH) free radical. These extracts showed an IC50 value of respectively 5, 20 and 65?µg?mL^?1. The beneficial effects of TOF, ethyl acetate and methanol extracts of C. rotundus were assessed by antioxidant and antimutagenic activities.     

Screening and bioassays for biologically-active substances from forty marine sponge species from San Diego,California, USA

Biologically-active substances were investigated from 35 demosponge species and 5 calcareous sponge species collected from intertidal or shallow subtidal habitats near San Diego, California, USA, from 1978 to 1980. Crude methanolic extracts of each species were tested for suppression of growth by bacteria (7 species) and a yeast. Antimicrobial activity was found in 26 Demospongiae and 2 Calcarea. Strong activity was found in 11 demosponges and, subsequently, 38 natural products with antimicrobial activity were isolated from 8 of these species (Aplysina fistularis, Dysidea amblia, Leiosella idia, Euryspongia sp., Toxadocia zumi, Axinella sp., Haliclona?cinerea and ?Pachychalina lunisimilis). Twenty-eight of these natural products (usually as pure compounds) were assayed for: (1) suppression of growth of marine fungi (3 spp.) and a red alga; (2) behavior modifications of invertebrate adults (4spp.); (3) toxicity to a goldfish; (4) inhibition of sexual reproduction of a brown alga; (5) inhibition of settlement and/or metamorphosis of late larvae or invertebrate juveniles (4spp.). Many of the natural products were also incorporated into pelleted fish food and tested for feeding-behavior modifications of fishes (5 spp.). Three of the compounds from Dysidea amblia were inactive in all tests. All other natural products were active in at least one assay, although none was active in all assays. The discussion relates the possession of biologically-active substances to the ecology of each sponge species; for example, sponges with antimicrobial substances are rarely overgrown.