Antibacterial activities in various tissues of the horse mussel, Modiolus modiolus

A search for antibacterial activity in different organs/tissues of the horse mussel, Modiolus modiolus, was conducted. Dried samples were extracted with 60% (v/v) acetonitrile, containing 0.1% (v/v) trifluoroacetic acid. Due to high salt content, two liquid phases were obtained; an acetonitrile-rich phase (ACN extract) and an aqueous phase. The aqueous phase was further subjected to solid phase extraction (SPE). Eluates from SPE and ACN extracts were tested for antibacterial, lysozyme, and toxic activity. Antibacterial activity was demonstrated in extracts from several tissues, including plasma, haemocytes, labial palps, byssus, mantle, and gills. Some of the extracts were sensitive to proteinase K treatment, indicating antibacterial peptides and/or proteins. Lysozyme-like activity and toxic activity against Artemia salina nauplii was detected in fractions from the gills, mantle, muscle, and haemocytes. Results from this study indicate that M. modiolus is a promising source for identifying novel drug lead compounds.     

Biodiversity of benthic invertebrates and organic matter processing in shallow marine sediments: an experimental study

The main objective of this study was to measure the impact of benthic invertebrate diversity on processes occurring at the water-sediment interface. We analyzed the effects of interactions between three shallow water species (Cerastoderma edule, Corophium volutator, and Nereis diversicolor). The impacts of different species richness treatments were measured on sediment reworking, bacterial characteristics, and biogeochemical processes (bromide fluxes, O₂ uptake, nutrient fluxes, and porewater chemistry) in sediment cores. The results showed that the three species exhibited different bioturbation activities in the experimental system: C. edule acted as a biodiffusor, mixing particles in the top 2 cm of the sediments; C. volutator produced and irrigated U-shaped tubes in the top 2 cm of the sediments; and N. diversicolor produced and irrigated burrow galleries in the whole sediment cores. C. edule had minor effects on biogeochemical processes, whereas the other species, through their irrigation of the burrows, increased the solute exchange between the water column and the sediment two-fold. These impacts on sediment structure and solute transport increased the O₂ consumption and the release of nutrients from sediments. As N. diversicolor burrowed deeper in the sediment than C. volutator, it irrigated a greater volume of sediments, with great impact on the sediment cores.Most treatments with a mixture of species indicated that observed values were often lower than predicted values from the addition of the individual effects of each species, demonstrating a negative interaction among species. This type of negative interaction measured between species on ecosystem processes certainly resulted from an overlap of bioturbation activities among the three species which lived and foraged in the same habitat (water-sediment interface). All treatments with N. diversicolor (in isolation and in mixture) produced similar effect on sediment reworking, water fluxes, nutrient releases, porewater chemistry, and bacterial characteristics. Whichever species associated with N. diversicolor, the bioturbation activities of the worm hid the effect of the other species. The results suggest that, in the presence of several species that use and modify the same sediment space, impact of invertebrates on ecosystem processes was essentially due to the most efficient bioturbator of the community (N. diversicolor). In consequence, the functional traits (mode of bioturbation, depth of burrowing, feeding behaviour) of an individual species in a community could be more important than species richness for some ecosystem processes.     

Screening marine algae from China for their antitumor activities

Thirty-nine species of marine algae collected from the coast of China were screened for their antitumor activities, and eight species Leathesia difformes, Polysiphonia urcedata, Scytosiphon lomentarius, Gloiopeliis furcata, Punctaria latifolia, Symphyocladia latiuscula, Rhodomela confervoides and Ulva pertusa showed potent cytotoxic activities. Three, Rhodomela confervoides, Scytosiphon lomentarius and Gloiopeliis furcata, were used for further investigation. More than 30 compounds were isolated and purified, and 14 bromophenols, 1 steroid and 1 carotene were identified by advanced spectroscopic methods including IR, MS, and NMR techniques. Amongst the 16 identified compounds, 7 showed vigorously selective activities against KB, Bel7402 and A549 cancer cells, and 6 bromophenols were new compounds.     

The role of chemical signals in the spawning induction of polychaete worms and other marine invertebrates

Extracts of gonad, body fluid and spawning water (into which gametes have been released by the process of spawning) were tested for the presence of spawning-inducing activity (SIA) on both sexes of the polychaetes Nereis succinea (Frey and Leuckhart) and Platynereis dumerilii (Audouin and Milne-Edwards). Gonadal and body fluid extracts from Asterias rubens (L.) and Echinus esculentus (L.) (echinoderms), Arenicola marina (L.) (polychaete), Clupea harengus (L.) (teleost) and body fluid and spawning water of Nereis virens (Sars) (polychaete) all exhibit SIA on male N. succinea. C. harengus extract from male gonads also had SIA on male P. dumerilii. Phytoplankton monoculture extracts were also tested. The cryptomonad (Rhodomonas baltica) (Karsten) had SIA on male P. dumerilii, and the diatom Phaeodactylum tricornutum (Bohlin) had some activity on Mytilus edulis (L.). The active fraction of R. baltica is chromatographically similar to uric acid (the natural sperm-release pheromone produced by females of P. dumerilii) [Nature 382 (1996) 214], but has a different retention time and UV absorbance spectra on reverse phase HPLC. Conspecific extracts of spawning water and gonadal material did not have SIA on the same and opposite sex in A. marina and N. virens. The role of chemical signals in marine invertebrates is discussed in relation to their mode of spawning.     

Biomedicinals from the phytosymbionts of marine invertebrates: a molecular approach

Marine invertebrate animals such as sponges, gorgonians, tunicates and bryozoans are sources of biomedicinally relevant natural products, a small but growing number of which are advancing through clinical trials. Most metazoan and anthozoan species harbour commensal microorganisms that include prokaryotic bacteria, cyanobacteria (blue-green algae), eukaryotic microalgae, and fungi within host tissues where they reside as extra- and intra-cellular symbionts. In some sponges these associated microbes may constitute as much as 40% of the holobiont volume. There is now abundant evidence to suggest that a significant portion of the bioactive metabolites thought originally to be products of the source animal are often synthesized by their symbiotic microbiota. Several anti-cancer metabolites from marine sponges that have progressed to pre-clinical or clinical-trial phases, such as discodermolide, halichondrin B and bryostatin 1, are thought to be products derived from their microbiotic consortia. Freshwater and marine cyanobacteria are well recognised for producing numerous and structurally diverse bioactive and cytotoxic secondary metabolites suited to drug discovery. Sea sponges often contain dominant taxa-specific populations of cyanobacteria, and it is these phytosymbionts (= photosymbionts) that are considered to be the true biogenic source of a number of pharmacologically active polyketides and nonribosomally synthesized peptides produced within the sponge. Accordingly, new collections can be pre-screened in the field for the presence of phytobionts and, together with metagenomic screening using degenerate PCR primers to identify key polyketide synthase (PKS) and nonribosomal peptide synthetase (NRPS) genes, afford a biodiscovery rationale based on the therapeutic prospects of phytochemical selection. Additionally, new cloning and biosynthetic expression strategies may provide a sustainable method for the supply of new pharmaceuticals derived from the uncultured phytosymbionts of marine organisms.     

Antimycobacterial,antibacterial and antifungal activities of Terminalia superba (Combretaceae)

The methanol extract from the stem bark of Terminalia superba (TSB), fractions (TSB1–7) and two compounds isolated following bio-assay guided fractionation namely 3,4′-di-O-methylellagic acid 3′-O-β-d-xylopyranoside (1) and 4′-O-galloy-3,3′-di-O-methylellagic acid 4-O-β-d-xylopyranoside (2) were evaluated for their antimycobacterial, antibacterial and antifungal activities. The broth microdilution, the microplate Alamar Blue assay (MABA) and the agar disc diffusion methods were used for the investigations. The results of the antimycobacterial assays showed that the crude extract, fractions TSB5–7 and compound 1 were able to prevent the growth of all the studied mycobacteria. The lowest minimal inhibitory concentration (MIC) value of 39.06 µg/ml for this extract was recorded on both M. smegmatis and M. tuberculosis MTCS2. The corresponding values were 19.53 µg/ml and 4.88 µg/ml for fractions and compounds respectively. The MIC determination results on other organisms indicated values ranging from 19.53 to 78.12 µg/ml for TSB and compound 2 on 90.9% of the tested organisms, meanwhile compound 1 as well as fractions TSB 6 and 7 exhibited detectable MIC values on all studied microorganisms. The overall results provide promising baseline information for the potential use of the crude extract from T. superba, fractions 6–7 and the tested compounds in the treatment of tuberculosis, bacterial and fungal infections.     

The control of the development of a marine benthic community by predation on recruits

Recruitment is an important process in regulating many marine benthic communities and many studies have examined factors controlling the dispersal and distribution of larval immigrants. However, benthic species also have early post-settlement life-stages that are dramatically different from adult and larval stages. Predation on these stages potentially impacts measured recruitment and the benthic populations and communities that ultimately develop.We examined the consequences of post-settlement predation on 1-day-old to 1-month-old recruits of sessile invertebrates at two field sites in southern New England. One site (Breakwater) was in a protected area with few predators and the other (Pine Island) was <1 km away in an open coast area with three different predator guilds: small and large invertebrates and fish. The Breakwater site had been dominated for >10 years by colonial and solitary ascidians. These species were absent from the Pine Island site which was dominated by bryozoans. Our goal was to examine whether post-settlement predation influenced the development and subsequent structure of the epifaunal community.Here we examine long-term changes in community development resulting from post-settlement predation, and contrast these results to those of earlier experiments examining the reductions in observed recruitment by post-settlement predation. Our first long-term experiment examined natural community development at the two sites and whether transplanted communities changed when exposed to the different levels of predation at these sites. The communities that developed at both sites were consistently different from each other and similar to resident communities at their respective sites. On panels transplanted from the Breakwater to Pine Island, solitary ascidians and the colonial ascidian, Botryllus schlosseri, suffered high mortalities on both caged and uncaged treatments, indicative of predation by small predators that could enter cages. Some solitary ascidians did survive inside cages and the colonial ascidian, Botrylloides violaceus, became dominant on all transplanted treatments. On panels transplanted from Pine Island to the Breakwater, ascidians invaded and dominated all treatments except those that were originally caged at Pine Island.In the second long-term experiment, natural communities were allowed to develop on panels exposed at the Breakwater for 1, 2, 3, and 4 weeks. Each set was transplanted to three treatments at Pine Island: open uncaged pilings, caged pilings to exclude fish and large invertebrates, and racks suspended above the bottom to exclude all predators. When 1-week-old communities were transplanted, after 2-3 weeks only bryozoans were found on the open and caged pilings, while colonial ascidians dominated the suspended rack treatment. When older 2-week-old communities were transplanted, colonial ascidians also became dominant in the caged piling treatment and when 3- and 4-week-old communities were transplanted colonial ascidians dominated all three treatments. Solitary ascidians were never abundant on open pilings exposed to fish and large benthic invertebrate predators.Post-settlement predator-prey interactions involved newly settled and juvenile life-stages of a variety of prey species and many invertebrate and vertebrate predator species. The effects of these interactions on recruitment did result in differences in the development and eventual species composition of the communities, even though predators had little if any effect on the adults of the prey species.     

Antimycobacterial,antibacterial and antifungal activities of the methanol extract and compounds from Thecacoris annobonae (Euphorbiaceae)

This study was designed to evaluate the antimycobacterial, antibacterial and antifungal activities of the methanol extract from the stem bark of Thecacoris annobonae Pax & K. Hoffm, that of aristolochic acid I (1) and other isolated compounds. The microplate alamar blue assay (MABA) and the broth microdilution method were used to determine the minimal inhibitory concentration (MIC) and minimal microbicidal concentration (MMC) of the above samples. The H⁺-ATPase-mediated proton pumping assay was used to evaluate a possible mechanism of action for both the methanol extract and aristolochic acid I. The results of the MIC determinations showed that the methanol extract and aristolochic acid I prevent the growth of all studied organisms. The results obtained in this study also showed that the methanol extract as well as aristolochic acid I inhibited the H⁺-ATPase activity. The overall results provided evidence that the methanol extract of T. annobonae might be a potential source of new antimicrobial drug against tuberculosis, and some bacterial and fungal diseases, but should be consumed with caution, bearing in mind that the main active component, aristolochic acid I is a potentially toxic compound.     

Biological activities of ACL-I and physicochemical properties of ACL-II, lectins isolated from the marine sponge Axinella corrugata

Lectin II from the marine sponge Axinella corrugata (ACL-II) was purified by affinity chromatography on rabbit erythrocytic stroma incorporated into a polyacrylamide gel, followed by gel filtration on Ultrogel AcA 44 column. Purified ACL-II is a lectin with an Mr of 80 kDa and 78 kDa, estimated by SDS-PAGE and by FPLC on Superose 12 HR column, respectively. ACL-II mainly agglutinates native rabbit erythrocytes and this hemagglutinating activity is independent of Ca^2 +, Mg^2 + and Mn^2 +, but is inhibited by d-galactose, chitin and N-acetyl derivatives, with the exception of GalNAc. ACL-II is stable for up to 65 °C for 30 min, with a better stability at a pH range of 2 to 6. In contrast, ACL-I displays a strong mitogenic and cytotoxic effect.     

The antibacterial effect of a polyhydroxylated fucophlorethol from the marine brown alga,Fucus vesiculosus

An antibacterial compound was isolated from the brown alga Fucus vesiculosus. Purification consisted of extraction of plant material with 0.05% trifluoroacetic acid, concentration on a C₁₈ cartridge, and reverse-phase high performance liquid chromatography on a C₁₈ semi-preparative column. The isolated compound exhibited antibacterial activity against both the Gram-positive and the Gram-negative bacteria tested. Killing studies conducted indicated that the activity was bactericidal. The compound showed no haemolytic effect against human red blood cells. Results obtained by electrospray ionization mass spectrometry indicated that the antibacterial activity was caused by a polyhydroxylated fucophlorethol.     

Penicisteroids A and B, antifungal and cytotoxic polyoxygenated steroids from the marine alga-derived endophytic fungus Penicillium chrysogenum QEN-24S

Two new polyoxygenated steroids, namely, penicisteroids A and B (1 and 2), were obtained from the culture extract of Penicillium chrysogenum QEN-24S, an endophytic fungus isolated from an unidentified marine red algal species of the genus Laurencia. In addition, seven known steroids (3-9) were also isolated and identified. The structures of these compounds were established on the basis of extensive spectroscopic analysis. The absolute configuration for compound 1 was determined by application of the modified Mosher’s method. Penicisteroid A (1), which is a structurally unique steroid having tetrahydroxy and C-16-acetoxy groups, displayed potent antifungal and cytotoxic activity in the preliminary bioassays. Preliminary structure-activity relationships are discussed.     

Antiplasmodial and antifungal activities of iridal,a plant triterpenoid

Iridal, a triterpenoidic compound extracted from Iris germanica L., was previously shown to have an interesting activity on two cultured human tumor cell lines (A2780 and K562). In the present work, this same product was tested in vitro on Plasmodium falciparum chloroquine-resistant and -sensitive strains, in vivo on P. vinckei, and on some Candida albicans and C. parapsilosis strains too. The IC(50) obtained in vitro on human malaria strain ranged from 1.8 to 26.0 microg/ml and the ED(50) in vivo is about 85 mg/kg/day by intraperitoneal route. The minimal inhibitory concentrations were higher than to 50 microg/ml, whatever the strain of yeast tested. This product presents an antiplasmodial activity similar to that obtained with extracts from the plant Azadirachta indica classically taken as reference in malaria phytomedicine. Conversely iridal shows no important antifungal activity. The specific activity of iridal on human malaria parasite and on tumor cell lines is discussed.     

Study on bioactivity of extracts from marine sponges in Chinese Sea

The bioactivity screening of fractions from two inter-tidal sponges collected from the north of China Yellow Sea and one sponge collected from the South Chinese Sea was reported in this study. In sponge Hymeniacidon perleve there were 9 fractions out of 15 from CHCl₃ extract with anti Staphylococcus aureus activity, 9 fractions out of 19 from BuOH extract with anti Escherichia coli activity, and three fractions from CHCl₃ extract which had moderate to strong activity in inhibiting Bacillus subtilis, Candida albicans, and Aspergilus niger. The fractions of Reniochalina sp. showed bioactivity against bacteria and fungi. The fractions of Acanthella acuta Schmidt showed bioactivity against S. aureus and fungi. One compound from H. perleve obtained by the bioactively directing isolation was tested for bioactivity against the human hepatoma cell line Qgy7701 (IC₅₀ 10.1 μg/ml), Burkitt's lymphoma cell line Raji (IC₅₀ 9.76 μg/ml) and chronic myelogenous leukemia K562 (IC₅₀ 1.90 μg/ml).     

Insect-antifeedant and antibacterial activity of diterpenoids from species of Plectranthus

Bio-assay guided fractionation of an acetone extract of leaf material from Plectranthus saccatus Benth. resulted in the isolation of a beyerane diterpenoid. This compound, characterised by spectroscopic methods as ent-3beta-(3-methyl-2-butenoyl)oxy-15-beyeren-19-oic acid, showed insect antifeedant activity against Spodoptera littoralis. Known quinonoid abietane diterpenoids obtained from new sources included a mixture of the (4R,19R) and (4R,19S) diastereoisomers of coleon A from P. aff. puberulentus J.K. Morton, coleon A lactone from P. puberulentus J.K. Morton, and coleon U and coleon U quinone from P. forsteri 'Marginatus' Benth. These compounds, and the crude acetone extracts from the leaf surfaces of 11 species of Plectranthus, were tested for antifeedant activity against S. littoralis, antibacterial activity against Bacillus subtilis and Pseudomonas syringae and antifungal activity against Cladosporium herbarum. The coleon A mixture showed potent antifeedant activity against S. littoralis, whereas coleon U showed the greatest antimicrobial activity.     

Chemical composition and antibacterial activities of the essential oils of Lippia chevalieri and Lippia multiflora from Burkina Faso

The chemical composition of the essential oils of Lippia chevalieri and Lippia multiflora obtained from the air-dried leaves by hydrodistillation were analysed using GC and GC-MS. L. chevalieri and L. multiflora belonged to thymol/p-cymene/2-phenyl ethyl propionate and thymol/p-cymene/thymyl acetate chemotypes, respectively. The essential oils were also tested against 09 strains using a broth microdilution method. The Gram-negative bacteria were the most sensitive. The essential oil of L. multiflora was the most active.     

Complexes of 2-acetyl-gamma-butyrolactone and 2-furancarbaldehyde thiosemicarbazones: antibacterial and antifungal activity

Cobalt, nickel, copper and zinc coordination compounds of two thiosemicarbazones with general composition ML₂ (L: monodeprotonated ligand corresponding to 2-acetyl-γ-butyrolactone thiosemicarbazone, HL¹, and 2-furancarbaldehyde thiosemicarbazone, HL²) and also complexes with general composition MCl₂(HL²) were synthesized (except [NiCl₂(HL²)] and [Co(L²)₂]). The interaction of CuCl₂ with HL² gave [CuCl(HL²)], a copper(I) complex. The ligands and metal complexes were characterized by IR, ¹H and ¹³C NMR spectroscopy, and magnetic susceptibility measurements. The crystal structure of [Ni(L²)₂] · 2dmso was determined and a trans-square planar coordination of the two κ²-N,S chelate rings forming polymeric strips through H-bonds with dmso was observed. Actually, in all the reported complexes both ligands behaved as κ²-N,S chelates, except in the case of [Co(L¹)₂] in which HL¹ is tridentate κ³-N,S,O. The antimicrobial properties of all compounds were studied using a wide spectrum of bacterial and fungal strains. The copper complexes of HL² were the most active against all strains, including dermatophytes and phytopathogenic fungi. Most of the studied compounds, especially [Cu(L¹)₂], presented good activity against Haemophilus influenzae, a very harmful bacterium to humans.     

Screening of Echinops ellenbeckii and Echinops longisetus for biological activities and chemical constituents

Members of the genus Echinops in the family of Asteraceae are widely used in Ethiopian herbal medicine for the treatment of various diseases and illness such as migraine, diarrhea, heart pain, different forms of infections, intestinal worm infestation and hemorrhoid. Hydroalcoholic extracts of the root, flower head, leaf and stem of Echinops ellenbeckii O. Hoffm. and Echinops longisetus A. Rich were investigated for their chemical constituents and biological activities. The presence of alkaloids, saponins, phytosterols, polyphenols and carotenoids in the different parts of the plants was observed whilst anthraquinones were not detected. The leaf extracts of both plants and stem extract of E. longisetus showed strong inhibitory activity against cultures of Staphylococcus aureus. None of the extracts were found to be active against Gram-positive organisms. The flower extract of E. ellenbeckii showed strong inhibitory activity against Candida albicans. Root and flower extracts of the plants investigated showed lethal activity against earthworms. Moreover, the extracts of the roots of both plants showed molluscicidal activity against schistosome-transmitting snail hosts. The biological activities observed were dose dependent.     

Thermal control of hatchling emergence patterns in marine turtles

The emergence patterns of green turtle (Chelonia mydas) hatchlings on two beaches on Ascension Island, South Atlantic were monitored and related to thermal patterns in the sand at 10, 20, 30 and 40 cm depth. A total of 6001 hatchlings were recorded emerging on Long Beach, and 3171 emerged on North East Bay during the study period. No significant difference was observed in the temporal pattern of hatchling emergence among nights, or between the two beaches. Hatchling emergence predominantly occurred at night with over 93% of hatchlings emerging during the hours of darkness. Almost all hatchlings emerging in daylight suffer predation by the Ascension frigatebird (Fregata aquila). Counts of frigatebirds both above the study beaches and offshore were highest just after sunrise, with a smaller peak prior to sunset, when frigatebirds were found to predate hatchlings emerging, crawling down the beach or detected in inshore waters. The likely thermal cues controlling hatchling emergence were investigated (temperature at different depths, thermal gradients in the sand and temperature change). The most plausible thermal factor appears to be the change of temperature at superficial sand depths, with hatchling emergence inhibited when subsurface sand temperatures were increasing. This simple mechanism is likely to ensure predominantly nocturnal hatchling emergence regardless of sand albedo, seasonality or latitude as long as night is relatively cooler than day.