Inhibition of pathogenic Vibrio harveyi using calamenene, derived from the Indian gorgonian Subergorgia reticulata, and its synthetic analog.

We report the synthesis and antimicrobial properties of a partially reduced dihydronathphthoquinone analogue of 2-methoxy, 5-acetoxy calamenene, extracted from Subergorgia reticulata. The growth of a pathogenic Vibrio harveyi strain was effectively controlled by the calamenene derivative 1 (Cala1) and its synthetic analog 2 (Cala2). Complete mortality of V. harveyi was observed with 2.5 and 0.5 µg mL-1 concentrations of Cala1 and Cala2, respectively. The metabolic assays demonstrated that Cala1 is a bacteriostatic agent while Cala2 showed bactericidal properties. It was confirmed that translocation of Cala2 into the cytoplasm does not induce any change to the integrity of the bacterial cell wall. The Cala2 induced damage to the genetic material of 70% of cells while genetic material of 91% of cells treated with Cala1 remained intact. The Cala2 is, therefore, proposed as a potential bactericidal compound against the aquaculture pathogen V. harveyi. The fact that the Cala2 exhibited minimal cytotoxicity to Artemia nauplii indicates its potential use as an antimicrobial agent for aquaculture operations.

Response of particle-associated bacteria to long-term heavy metal contamination in a tropical estuary.

Estuaries being the connecting link between terrestrial and marine environment, experience spatial variations in the hydrographic variables as well as concentrations of pollutants. The present study reports a contrasting difference in the metal tolerance and enzyme activity of particle-associated bacteria (PAB) isolated from the upstream and downstream reaches of a tropical estuary [Cochin Estuary (CE) in the southwest coast of India], exposed to different levels of heavy metal contamination. The upstream of the estuary has been overloaded with heavy metals in the last few decades, while the downstream is less polluted. There were only 25% of culturable PAB phylogenetically common in both upstream and downstream. The PAB isolated from the upstream were dominated by γ-proteobacteria (48.1%) followed by α-proteobacteria (25.0%), while it was in the reverse order of α-proteobacteria (45.9%) and γ-proteobacteria (36.1%) in the downstream. More number of PAB from the upstream showed tolerance to higher concentrations of Zn and Cd. The Acinetobacter sp. MMRF1051 isolated from the upstream showed tolerance up to 250 mM Zn, 100 mM Cd, and 250 mM Ni. The enzyme expression profile of PAB from downstream was in the order of lipase > phosphatase > ß-glucosidase > aminopeptidase, while it was in the order of ß-glucosidase > lipase > aminopeptidase > phosphatase in the upstream of the estuary. The present study shows the selective pressure exerted by heavy metal pollution on the diversity of culturable bacteria associated with particulate matter in a tropical estuary. Also, the variation in their enzyme activities may impinge the remineralization of particulate organic matter (POM) in the system and may impart adverse impacts on ecosystem functioning.

Role of heavy metals in structuring the microbial community associated with particulate matter in a tropical estuary.

Particulate matter (PM), which are chemically and biochemically complicated particles, accommodate a plethora of microorganisms. In the present study, we report the influence of heavy metal pollution on the abundance and community structure of archaea and bacteria associated with PM samples collected from polluted and non-polluted regions of Cochin Estuary (CE), Southwest coast of India. We observed an accumulation of heavy metals in PM collected from CE, and their concentrations were in the order Fe > Zn > Mn > Cr > Pb > Cu > Cd > Co > Ni. Zinc was a major pollutant in the water (4.36-130.50 µgL-1) and in the particulate matter (765.5-8451.28 µgg-1). Heavy metals, Cd, Co, and Pb were recorded in the particulate matter, although they were below detectable limits in the water column. Statistical analysis showed a positive influence of particulate organic carbon, nitrogen, PM-Pb, PM-Zn and PM-Fe on the abundance of PM-archaea and PM-bacteria. The abundance of archaea and bacteria were ten times less in PM compared with planktonic ones. The abundance of PM-archaea ranged between 4.27 and 9.50 × 107and 2.73 to 3.85 × 107 cellsL-1 respectively for the wet and dry season, while that of PM-bacteria was between 1.14 and 6.72 × 108 cellsL-1 for both seasons. Community structure of PM-bacteria varied between polluted and non-polluted stations, while their abundance does not show a drastic difference. This could be attributed to the selective enrichment of bacteria by heavy metals in PM. Such enrichment may only promote the growth of metal resistant archaea and bacteria, which may not participate in the processing of PM. In such cases, the PM may remain without remineralization in the system arresting the food web dynamics and biogeochemical cycles.

Diversity and bioactive potentials of culturable heterotrophic bacteria from the surficial sediments of the Arabian Sea.

Marine sediments accommodate plethora of diverse microorganisms with varying ecological functions. In the present study, we isolated bacteria from surficial sediments of south east Arabian Sea (AS) and evaluated their bioactive potentials. A total of 131 isolates belonging to the phylum: γ-Proteobacteria (63%), Bacillales (34%) and Micrococcaceae (3%) were isolated. Among these, about 40% of the isolates showed the presence of secondary metabolite biosynthetic genes such as PKS or NRPS or both. Organic extracts of nearly 50% of these organisms were cytotoxic to human breast cancer MCF-7 cells and were bactericidal to human pathogens, Escherichia coli and Pseudomonas sp., while 20-30% of them were bactericidal to Vibrio sp. and Staphylococcus sp. too. In all, 8 isolates, belonging to Pseudomonas spp., Bacillus sp. and/or Lysinibacillus sp. displayed high level of bactericidal/cytotoxic properties. The study proposes AS sediment as a rich source for microorganisms with prospective bioactive molecules.

Recent advances in marine drug research.

Structures and properties of promising marine anti-cancer, anti-inflammation and anti-infectious (HIV, HSV, malaria, leishmania) compounds reported during 2008-2011 are discussed. Wherever possible, attempts have also been made to highlight their possible biogenesis or structure-activity relationships (SAR). Since the stress is on identifying and short-listing potential drug molecules, this review is restricted to only those compounds exhibiting promising in vitro activity, the arbitrary cut off being IC50<15 µM, reported during the above period.

Surface topographical and ultrastructural alterations of Raillietina echinobothrida and Ascaridia galli induced by a compound isolated from Acacia oxyphylla.

The stem bark of Acacia oxyphylla Graham ex Bentham is used as an anthelmintic by the natives of Mizoram (North-East India). Therefore, the aim of the study was to assess the effect of the active compound isolated from A. oxyphylla on the tegument of adult Raillietina echinobothrida and Ascaridia galli. The test parasites R. echinobothrida and A. galli were incubated in physiological buffered saline containing 0.0005, 0.001, 0.05, 0.1 and 1mg/ml of the isolated compound. The alterations in the tegument of the parasites post paralysis were examined using electron microscopes. The compound reduced the cestode's motility soon after incubation, but did not induce paralysis in the nematodes till about 11-14 h at highest concentration. The compound caused extensive digestion of cestode tegument as evident by electron microscopy. Disorganization of muscle bundles, loss of cell-cell contact, extreme vacuolization and oedema were some of the changes observed. Loss of cellular organelles combined with distortion of those present was markedly noted throughout the parasite tissue. Deformation and disorganization of epicuticle, disruption of mitochondrial and nuclear membrane were also observed in nematode exposed to the active compound of the plant. Substantial structural deformities in the treated parasites are indicative of an efficient vermicidal activity of the isolated compound against cestodes and nematodes.

Calamenenes--aromatic bicyclic sesquiterpenes--from the Indian gorgonian Subergorgia reticulata (Ellis and Solander, 1786).

Three aromatic sesquiterpene derivatives, (+)-(7R, 10S)-2-methoxy calamenene (1), (+)-(7R, 10S)-2,5-dimethoxy calamenene (2) and (+)-(7R, 10S)-2-methoxy-5-acetoxy calamenene (3) were isolated from the methanol extract of the Indian gorgonian Subergorgia reticulata. Compound 2 has not been previously described in the literature. Compound 3 has not been isolated previously from a natural source. Compounds 1, 2 and 3 showed settlement inhibition activity against cyprids of Balanus amphitrite with EC(50) values of 4.4, 7.8 and 0.03 µg mL(-1), respectively. These compounds also exhibited considerable activity against Artemia nauplii at 50 µg mL(-1), indicative of their potential use as cytotoxic agents.

Active pharmaceutical ingredient (api) from an estuarine fungus, Microdochium nivale (Fr.).

Various marine habitats sustain variety of bio-sources of ecological and biotech potentials. Pharmaceutical potential compound Cyclosporine A was reported from marine fungus Microdochium nivale associated with Porteresia coarctata, a marine salt marsh grass from mangrove environment distributed along the Central West Coast (CWC) of India. This study involves association of M. nivale with P. coarctata plant, fermentation conditions, purification of Cyclosporine A, chemical characterization etc. Its antifungal inhibition and MIC (Minimum inhibitory concentration) against Aspergillus strains (A. niger, A. japonicus, A. fresenii), yeasts and dermatophytes (Candida sp., Cryptococcus neoformans, Trichophyton mentagrophytes, T. tonsurans, T. violaceum, Microsporium gypsum and Fusarium sp.) were evaluated. However, the MIC against A. japonicus, C. neoformans, Candida sp. and T. tonsurans were confirmed to be as low as 12.5-25 mg disc(-1). The antifungal properties of Cyclosporine A against Aspergillus species, yeast and dermatophytes revealed that CyclosporineAwould be a potential compound for life threatening diseases caused by above fungi in both human and animals. Furthermore, we have reported herewith another source of Cyclosporin Aderived from filamentous fungus, M. nivale. occurring in marine environment.

Two selective novel triterpene glycosides from sea cucumber, Telenata Ananas: inhibitors of chemokine receptor-5.

The aqueous methanolic extract of a sea cucumber was found to contain two triterpene glycosides 1 and 2. The structures of 1 and 2 were established based on high-resolution NMR studies. Compounds 1 and 2 exhibited inhibitory activity (K(i)) of 30 and 5microM, respectively, in a chemokine receptor subtype 5 (CCR5) assay. Both compounds did not show any significant inhibition in a CXCR2 assay at 50microM, suggesting their selectivity for the CCR5 receptor.

Studies on the secondary metabolites from the Indian gorgonian Subergorgia suberosa: isolation and characterization of four analogues of the cardiotoxin subergorgic acid.

Chemical investigation of the methanol extract of the Indian ocean gorgonian coral Subergorgia suberosa resulted in isolation and identification of four novel compounds 2-5. Structural investigation revealed compound 1 to be subergorgic acid and the others its analogues.