George Seghal Kiran

Antimicrobial activity of seaweeds extracts against multiresistant pathogens

Fourteen seaweeds collected from the intertidal zone of Southwest coast of India were tested against ten human pathogen bacteria and one human pathogen fungus using the well diffusion test in the casitone agar medium. The species used in the present study include five Chlorophyta (Bryopsis plumosa, Ulva fasciata, Acrosiphonia orientalis, Chaetomorpha antennina, Grateloupia filicina), five Rhodophyta (Hypnea pannosa, Gracilaria corticata, Centroceras clavulatum, Portieria hornemannii, Cheilosporum spectabile) and four Phaeophyta (Padina tetrastromatica, Sargassum wightii, Stocheospermum marginatum, Chnoospora bicanaliculata). Of these, seven species were determined to be highly bioactive and screened on the multiresistant pathogens. We found that drying process has eliminated the active principles in the seaweeds. In the present study, methanol:toluene (3∶1) was found to be the best solvent for extracting the antimicrobial principles from fresh algae. However, the ethanolic extract showed no antibacterial activity.Acrosiphonia orientalis showed activity against 70% of the tested organisms.Stocheospermum marginatum was the only seaweed that showed activity againstKlebsiella pneumoniae. The extract fromGracilaria corticata was highly active againstProteus mirabilis, a Gram negative pathogenic bacterium. The present findings revealed that the tested seaweeds were highly active against Gram negative bacteria than Gram positive bacteria. The antimicrobial principle from seaweed was found to be a lipophilic compound. The compound was stable over a wide range of temperature (30–60 °C). The active principles of highly active seaweedsAcrosiphonia orientalis andStocheospermum marginatum were bactericidal.

Functional metagenomic strategies for the discovery of novel enzymes and biosurfactants with biotechnological applications from marine ecosystems.

Marine ecosystems are home to bacteria which are exposed to a wide variety of environmental conditions, such as extremes in temperature, salinity, nutrient availability and pressure. Survival under these conditions must have necessitated the adaptation and the development of unique cellular biochemistry and metabolism by these microbes. Thus, enzymes isolated from these microbes have the potential to possess quite unique physiological and biochemical properties. This review outlines a number of function‐based metagenomic approaches which are available to screen metagenomic libraries constructed from marine ecosystems to facilitate the exploitation of some of these potentially novel biocatalysts. Functional screens to isolate novel cellulases, lipases and esterases, proteases, laccases, oxidoreductases and biosurfactants are described, together with approaches which can be employed to help overcome some of the typical problems encountered with functional metagenomic‐based screens.

Biofilm disruption potential of a glycolipid biosurfactant from marine Brevibacterium casei.

The antibiofilm activity of a glycolipid biosurfactant isolated from the marine actinobacterium Brevibacterium casei MSA19 was evaluated against pathogenic biofilms in vitro. The isolate B. casei MSA19 was a potential biosurfactant producer among the 57 stable strains isolated from the marine sponge Dendrilla nigra. The biosurfactant production was optimized under submerged fermentation. The purified glycolipid showed a broad spectrum of antimicrobial activity. Based on the minimum inhibitory concentration/minimum bactericidal concentration ratio, the glycolipid was determined as bacteriostatic. The glycolipid biosurfactant disrupted the biofilm formation under dynamic conditions. The disruption of the biofilm by the MSA19 glycolipid was consistent against mixed pathogenic biofilm bacteria. Therefore, the glycolipid biosurfactant can be used as a lead compound for the development of novel antibiofilm agents.

Antimicrobial potential and seasonality of red algae collected from the southwest coast of India tested against shrimp, human and phytopathogens

Fifteen seaweeds belong to 13 families and 6 orders of the rhodophyta were sampled for one year from April 2007 to March 2008 along the southwest coast of India (Indian Ocean). The species were examined forin vitro antimicrobial activity against six pathogenicVibrio strains isolated from moribund tiger shrimp (Penaeus monodon), six type cultures (Microbial Type Culture Collection, MTCC) of prominent shrimpVibrio pathogens, 10 multidrug resistant clinical pathogens, four species ofCandida obtained from pulmonary TB patients and four species of plant pathogenic fungi to evaluate their potency to be used as natural antibiotics in pharmaceutical and agriculture field. Bioactivity was analyzed from crude extract of fresh and dried samples prepared from different polar and nonpolar solvents. Of these, four species of red algae (Asparagopsis taxiformis, Laurencia ceylanica, Laurencia brandenii, Hypnea valentiae) were found to be highly active. Broadest and highest activity was observed in the crude extract ofA. Taxiformis. Among the pathogens tested, shrimp pathogenicVibrios were the most susceptible organisms while phytopathogens were found to be little resistant. In the present study, methanol was found to be the best solvent for extracting antimicrobial metabolites from dried samples rather than fresh. Seasonal variation in the antimicrobial activity was observed with higher level of activity recorded fromA. Taxiformis between December and January. The active principle ofA. Taxiformis was purified in column chromatography, TLC and reverse phase HPLC. The individual HPLC peaks were subsequently tested against a panel of pathogenic microorganisms and the active constituent was identified by GC-MS. The antimicrobial profile ofA. Taxiformis suggested that lipophilic compound which was primarily composed of pyrrole-2-carboxylic acid, pentadecanoic acid and octadecanoic acid might have functional role in the chemical defence against microbial invasion and these compounds could be utilized for the development of medically potential products.

Bioactivity of the red algae Asparagopsis taxiformis collected from the Southwestern coast of India

Entre as diversas variedades de algas vermelhas, Asparogopsis taxiformis constitui uma das que apresentam alta biomassa na costa de Kollam (Sudoeste da India). No presente estudo A. taxiformis foi coletada, seca e reduzida a po, apos o que foi realizada sua extracao e feito o fracionamento usando-se cromatografia por coluna. As fracoes individuais foram avaliadas in vitro em ensaios para testar sua capacidade anti-incrustante, anticianobacteria e toxicidade para peixes e crustaceos. A fracao extraida com eter de petroleo e etil acetato (2:8) apresentou o espectro de bioatividade mais forte e amplo. No ensaio anti-incrustacao efetuado com com o molusco pulmonado Limnea truncatula, a fracao algal ativa produziu 80% de repelencia do pe em 150 mg/l, enquanto que no ensaio anticianobacteria a fracao ativa inibiu 100% do crescimento de Trichodesmium sp., em 320 mg/l. A fracao algal mostrou o efeito mais intenso contra peixes no nivel de 60 mg/l. Em relacao aos crustaceos, a toxicidade da fracao ativa foi avaliada tambem visando encontrar compostos nao toxicos para organismos nao alvo, tais como Penaeus monodon e Macrobrachium rosembergii. Foi visto que a fracao ativa da coluna mostrou menor toxicidade para estas especies. Os componentes quimicos da fracao ativa foram identificados por meio dos sistemas cromatograficos, tais como TLC, fase reversa do HPLC e GC-MS. O perfil geral de atividade aponta que a fracao ativa da coluna para A. taxiformis pode conter metabolitos bioativos sinergicos que podem ser utilizados para o controle de organismos incrustantes, explosao algal e peixes herbiboros/predadores em tanques de aquicultura.

Rhamnolipid biosurfactants: evolutionary implications, applications and future prospects from untapped marine resource

Abstract Rhamnolipid-biosurfactants are known to be produced by the genus Pseudomonas, however recent literature reported that rhamnolipids (RLs) are distributed among diverse microbial genera. To integrate the evolutionary implications of rhamnosyl transferase among various groups of microorganisms, a comprehensive comparative motif analysis was performed amongst bacterial producers. Findings on new RL-producing microorganism is helpful from a biotechnological perspective and to replace infective P. aeruginosa strains which ultimately ensure industrially safe production of RLs. Halotolerant biosurfactants are required for efficient bioremediation of marine oil spills. An insight on the exploitation of marine microbes as the potential source of RL biosurfactants is highlighted in the present review. An economic production process, solid-state fermentation using agro-industrial and industrial waste would increase the scope of biosurfactants commercialization. Potential and prospective applications of RL-biosurfactants including hydrocarbon bioremediation, heavy metal removal, antibiofilm activity/biofilm disruption and greener synthesis of nanoparticles are highlighted in this review.

Antiadhesive activity of poly-hydroxy butyrate biopolymer from a marine Brevibacterium casei MSI04 against shrimp pathogenic vibrios.

BackgroundVibrio pathogens are causative agents of mid-culture outbreaks, and early mortality syndrome and secondary aetiology of most dreadful viral outbreaks in shrimp aquaculture. Among the pathogenic vibrios group, Vibrio alginolyticus and V. harveyi are considered as the most significant ones in the grow-out ponds of giant black tiger shrimp Penaeus monodon in India. Use of antibiotics was banned in many countries due to the emergence of antibiotic-resistant strains and accumulation of residual antibiotics in harvested shrimp. There is an urgent need to consider the use of alternative antibiotics for the control of vibriosis in shrimp aquaculture. Biofilm formation is a pathogenic and/or establishment mechanism of Vibrio spp. This study aims to develop novel safe antibiofilm and/or antiadhesive process using PHB to contain vibrios outbreaks in shrimp aquaculture.ResultsIn this study a poly-hydroxy butyrate (PHB) polymer producing bacterium Brevibacterium casei MSI04 was isolated from a marine sponge Dendrilla nigra and production of PHB was optimized under submerged-fermentation (SmF) conditions. The effect of carbon, nitrogen and mineral sources on PHB production and enhanced production of PHB by response surface methods were demonstrated. The maximum PHB accumulation obtained was 6.74 g/L in the optimized media containing 25 g/L starch as carbon source, 96 h of incubation, 35°C and 3% NaCl. The highest antiadhesive activity upto 96% was recorded against V. vulnificus, and V. fischeri, followed by 92% against V. parahaemolyticus and V. alginolyticus and 88% inhibition was recorded against V. harveyi.ConclusionIn this study, a thermostable biopolymer was chemically characterized as PHB based on 1HNMR spectra, FT-IR and GC-MS spectra. The NMR spectra revealed that the polymer was an isocratic homopolymer and it also confirmed that the compound was PHB. The antiadhesive activity of PHB was determined in microtitre plate assay and an effective concentration (EC) of PHB (200 μl containing 0.6 mg PHB) was confirmed by confocal laser scanning microscopic analysis of vibrio biofilm on pre-treated glass and polystyrene surfaces. This is a first report on anti-adhesive activity of PHB against prominent vibrio pathogens in shrimp aquaculture.

Efficacy of marine green alga Ulva fasciata extract on the management of shrimp bacterial diseases

Secondary metabolites of the green algae, Ulva fasciata, were tested to determine the efficacy of controlling shrimp bacterial pathogens. Exploratory experiments indicated that an intermediate dose (1 g kg -1 of shrimp) of Ulva in the diet was highly effective at controlling bacterial pathogens of shrimp, as compared to lower (500 mg kg -1 ) and higher (1.5 g kg -1 ) doses. The pilot experiments evaluated the percent of relative protection afforded shrimps treated with Ulva diet and faced with various concentrations of bacterial pathogen. The survival of shrimps treated with Ulva diet was significant (P < 0.01). The present findings indicate that the green U. fasciata may be an excellent source for developing a potent medicated feed for shrimp disease management.

A halotolerant thermostable lipase from the marine bacterium Oceanobacillus sp. PUMB02 with an ability to disrupt bacterial biofilms

A halotolerant thermostable lipase was purified and characterized from the marine bacterium Oceanobacillus sp. PUMB02. This lipase displayed a high degree of stability over a wide range of conditions including pH, salinity, and temperature. It was optimally active at 30 °C and pH 8.0 respectively and was stable at higher temperatures (50–70 °C) and alkaline pH. The molecular mass of the lipase was approximately 31 kDa based on SDS-PAGE and MALDI-ToF fingerprint analysis. Conditions for enhanced production of lipase by Oceanobacillus sp. PUMB02 were attained in response surface method-guided optimization with factors such as olive oil, sucrose, potassium chromate, and NaCl being evaluated, resulting in levels of 58.84 U/ml being achieved. The biofilm disruption potential of the PUMB02 lipase was evaluated and compared with a marine sponge metagenome derived halotolerant lipase Lpc53E1. Good biofilm disruption activity was observed with both lipases against potential food pathogens such as Bacillus cereus MTCC1272, Listeria sp. MTCC1143, Serratia sp. MTCC4822, Escherichia coli MTCC443, Pseudomonas fluorescens MTCC1748, and Vibrio parahemolyticus MTCC459. Phase contrast microscopy, scanning electron microscopy, and confocal laser scanning microscopy showed very effective disruption of pathogenic biofilms. This study reveals that marine derived hydrolytic enzymes such as lipases may have potential utility in inhibiting biofilm formation in a food processing environment and is the first report of the potential application of lipases from the genus Oceanobacillus in biofilm disruption strategies.

Effect of Fe nanoparticle on growth and glycolipid biosurfactant production under solid state culture by marine Nocardiopsis sp. MSA13A

BackgroundIron is an essential element in several pathways of microbial metabolism, and therefore low iron toxicity is expected on the usage of Fe nanoparticles (NPs). This study aims to determine the effect of Fe NPs on biosurfactant production by marine actinobacterium Nocardiopsis sp. MSA13A under solid state culture. Foam method was used in the production of Fe NPs which were long and fiber shaped in nature.ResultsThe SEM observation showed non toxic nature of Fe NPs as no change in the morphology of the filamentous structure of Nocardiopsis MSA13A. The production of biosurfactant by Nocardiopsis MSA13A under solid state culture supplemented with Fe NPs increased to 80% over control. The biosurfactant produced by Nocardiopsis MSA13A was characterized as glycolipid derivative which effectively disrupted the pre-formed biofilm of Vibrio pathogen.ConclusionThe use of metal NPs as supplement would reduce the impact of non-metallic ions of the metal salts in a fermentation process. This would ultimately useful to achieve greener production process for biosurfactants. The present results are first report on the optimization of biosurfactant production under SSC using Fe NPs.