Ravi Kumar Asthana

Metal adsorption and desorption by lyophilized Pseudomonas aeruginosa

Biosorption of nickel (Ni2+) and copper (Cu2+) by lyophilized Pseudomonas aeruginosa cells was investigated based on Freundlich isotherm. Bacterial biomass showed significant sorption of both Ni (265 mg g−1) or Cu (137.6 mg g−1), and was also superior over the cation exchanger, IRA 400 (98 mg Ni g−1 or 26.6 mg Cu g−1). Metal binding by the test organism was a fast saturating, pH-dependent process. The optimum pH for Cu adsorption was 7.0 and for Ni 8.0. X-ray diffraction studies revealed that both cations were deposited on the cell predominantly as phosphide crystals. The participation of carboxyl, carbonyl, and phosphoryl groups along with H-bonding in metal sorption was evident in IR spectra. Biomass pretreatment by agents like NaOH, NH4OH or toluene enhanced the metal loading capacity, whereas, oven heating (80°C), autoclaving (120°C, 15 lb (in.2)−1), acid, detergent and acetone treatments were inhibitory. In bimetallic combination, Na, K or Ca increased sorption of Ni as well as Cu in contrast to Cd or Pb. Mineral acids (HCl, H2SO4 and HNO3) and NTA could recover more than 75% (on average) Ni or Cu adsorbed on the biomass. Calcium carbonate (10 mM) was efficient in Ni desorption (71%) compared to Cu (57%). Noticeably sodium carbonate remained specific for Cu remobilization (88%) than Ni (21%). The data are in favour of deployment of the test biomass as an efficient metal removal/recovery system.

Gene Expression in the Cyanobacterium Anabaena sp. PCC7120 under Desiccation

The N2-fixing cyanobacterium Anabaena sp. PCC7120 showed an inherent capacity for desiccation tolerance. A DNA microarray covering almost the entire genome of Anabaena was used to determine the genome-wide gene expression under desiccation. RNA was extracted from cells at intervals starting from early to late desiccation. The pattern of gene expression in DNA fragments was categorized into seven types, which include four types of up-regulated and three types of down-regulated fragments. Validation of the data was carried out by RT-PCR on selected up-regulated DNA fragments and was consistent with the changes in mRNA levels. Our conclusions regarding desiccation tolerance for Anabaena sp. PCC7120 are as follows: (i) Genes for osmoprotectant metabolisms and the K+ transporting system are up-regulated from early to mid-desiccation; (ii) genes induced by osmotic, salt, and low-temperature stress are up-regulated under desiccation; (iii) genes for heat shock proteins are up-regulated after mid-desiccation; (iv) genes for photosynthesis and the nitrogen-transporting system are down-regulated during early desiccation; and (v) genes for RNA polymerase and ribosomal protein are down-regulated between the early and the middle phase of desiccation. Profiles of gene expression are discussed in relation to desiccation acclimation.

Recent trends in development of biosensors for detection of microcystin.

Increased cyanobacterial blooms, a source of cyanotoxins are linked with climate change and eutrophication in aquatic bodies, a major concern worldwide. Microcystins are potently hepatotoxic, nephrotoxic as well as carcinogenic. Thus microcystins are threat to tourism, agriculture and animals health. However, there is a still lacuna in the knowledge of regulation of microcystins production. Presence of toxic and non-toxic cyanobacterial strains together and occurrence of various microcystin variants in aquatic bodies compounded the problem. Although several analytical techniques for microcystin detection such as bioassay, ELISA, HPLC and LC-MS etc. have been already prevalent, the development of biosensors offered rapid and accurate detection, high reproducibility and portability. Sequencing of Microcystis spp., opened the new vistas towards the development of biosensor at molecular and genetic level. This review incorporates the current trends in the development of biosensors for microcystin detection in the light of state-of-the-art techniques.

Identification of an antimicrobial entity from the cyanobacterium Fischerella sp. isolated from bark of Azadirachta indica (Neem) tree

The active principle in a methanolic extract of the laboratory-grown cyanobacterium, Fischerella sp. isolated from Neem (Azadirachta indica) tree bark was active against Mycobacterium tuberculosis, Enterobacter aerogenes, Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella typhi, Escherichia coli as well as three multi-drug resistant E. coli strains in in vitro assays. Based on MS, UV, IR 1H NMR analyses the active principle is proposed to be hapalindole T having the empirical formula C21H23N2ClSO and a molecular weight of 386 with the melting point range 179–182 °C. The estimated production of Hapalindole T from the cyanobacterium is 1.25 mg g−1 lyophilized biomass. It is suggested that cyanobacteria colonizing specialized niches such as tree bark could be an antibacterial drug resource.

Monitoring Approaches for a Toxic Cyanobacterial Bloom

Cyanobacterial blooms, dominated by Microcystis sp. and associated microcystin variants, have been implicated in illnesses of humans and animals. Little is known regarding the formation of blooms and the presence of cyanotoxin variants in water bodies. Furthermore, the role played by ecological parameters, in regulating Microcystis blooms is complicate and diverse. Local authorities responsible for water management are often faced with the challenging task of dealing with cyanobacterial blooms. Therefore, the development of suitable monitoring approaches to characterize cyanobacterial blooms is an important goal. Currently, various biological, biochemical and physicochemical methods/approaches are being used to monitor cyanobacterial blooms and detect microcystins in freshwater bodies. Because these methods can vary as to the information they provide, no single approach seemed to be sufficient to accurately monitor blooms. For example, immunosensors are more suited for monitoring the presence of toxins in clear water bodies while molecular methods are more suited to detect potentially toxic strains. Thus, monitoring approaches should be tailored for specific water bodies using methods based on economic feasibility, speed, sensitivity and field applicability. This review critically evaluates monitoring approaches that are applicable to cyanobacterial blooms, especially those that focus on the presence of Microcystis, in freshwater bodies. Further, they were characterized and ranked according to their cost, speed, sensitivity and selectivity. Suggested improvements were offered as well as future research endeavors to accommodate anticipated environmental changes.

Biohydrogen production from sugarcane bagasse by integrating dark- and photo-fermentation.

Hydrogen production from sugarcane bagasse (SCB) by integrating dark-fermentation by Enterobacter aerogenes MTCC 2822 and photo-fermentation by Rhodopseudomonas BHU 01 was investigated. The SCB was hydrolysed by sulphuric acid and the hydrolysate detoxified by passing through adsorbent resin column (Amberlite XAD-4) to remove the inhibitory furfural, and subjected to dark-fermentation. The cellulosic residue from acid hydrolysis was hydrolysed by the new isolate Cellulomonas fimi to release sugars for H2 production by E. aerogenes, through simultaneous saccharification, filtration and fermentation (SSFF). Cumulative H2 production during dark-fermentation and SSFF was 1000 and 613 ml/L, respectively. The spent media of dark-fermentation and SSFF were utilized for photo-fermentation by Rhodopseudomonas BHU 01. The cumulative H2 production was 755 ml/L for dark-fermentation and 351 ml/L for SSFF spent medium.

Copper uptake and its compartmentalization in Pseudomonas aeruginosa strains: Chemical nature of cellular metal

Copper-sensitive (Cus) and copper-resistant (Cur) strains of Pseudomonas aeruginosa were characterized in terms of Cu2+ sensitivity, uptake and its compartmentalization in the possible cell sectors. Minimum inhibitory concentrations (MICs) of Cu2+ for the Cur strain (3.2 mM and 0.12 mM in enriched- and in minimal-medium, respectively) were almost 5-fold higher over that of its sensitive counterpart. While Cus strain accumulated Cu2+ to a maximum of 1.8 μ mol mg−1 protein, Cur strain increased it to 2.37 μmol mg−1 protein. Both the strains also demonstrated energy- and pH-dependent Cu2+ uptake through the broad-substrate range divalent cation (Zn2+, Mg2+, Co2+) uptake system as well as through the system specific for Cu2+. Cell-fractionation study revealed that in Cur strain, periplasm and membrane are the main Cu2+ binding sites, whereas, in case of Cus strain, it is the cytoplasm. The overall observations indicate that the Cur strain restricted Cu2+ sequestration exterior to the cytoplasm as the possible strategy for Cu-resistance. The chemical nature of Cu2+ deposition in the respective strains was also ascertained by X-ray powder diffraction analysis.

Dynamics of microcystin production and quantification of potentially toxigenic Microcystis sp. using real-time PCR

Cyanobacterial blooms in eutrophied water body are generally composed of various genotypes with or without microcystin-producing genes (mcy gene cluster). Thus there is a need for quantification of potent toxin producing strains. The present study aimed at identifying microcystin variants and its producer strains in Durgakund pond, Varanasi, India, based on quantification of cpcBA-IGS and mcyA (condensation domain) genes using real-time PCR and LC-MS. Increase in microcystin concentrations was correlated with increase in mcyA copy number and the level of pigments (chlorophyll a, phycocyanin and carotenoids). Also, selected environmental factors (water temperature, light irradiance, rainfall, pH, N and P) and the concentration of microcystin variants (MC-LR, -RR and -YR) were also assessed in samples during May 2010 to April 2011 to establish the possible correlation among these parameters. Nutrients favored cyanobacterial bloom but it could not be correlated with the levels of microcystin variants and seemed to be geographically specific. Microcystis sp. dominant in the pond comprised potentially toxigenic cells. The ratio of potentially toxigenic Microcystis sp. to that of total Microcystis sp. ranged from 0% to 14%. Such studies paved the way to identify and quantify the most potent microcystin producer in the tropical aquatic body.

Status, Alert System, and Prediction of Cyanobacterial Bloom in South Korea

Bloom-forming freshwater cyanobacterial genera pose a major ecological problem due to their ability to produce toxins and other bioactive compounds, which can have important implications in illnesses of humans and livestock. Cyanobacteria such as Microcystis, Anabaena, Oscillatoria, Phormidium, and Aphanizomenon species producing microcystins and anatoxin-a have been predominantly documented from most South Korean lakes and reservoirs. With the increase in frequency of such blooms, various monitoring approaches, treatment processes, and prediction models have been developed in due course. In this paper we review the field studies and current knowledge on toxin producing cyanobacterial species and ecological variables that regulate toxin production and bloom formation in major rivers (Han, Geum, Nakdong, and Yeongsan) and reservoirs in South Korea. In addition, development of new, fast, and high-throughput techniques for effective monitoring is also discussed with cyanobacterial bloom advisory practices, current management strategies, and their implications in South Korean freshwater bodies.

Investigations on nickel biosorption and its remobilization

Abstract Immobilized preparations of the bacteria ( Pseudomonas aeruginosa and Rhodopseudomonas BHU strain 1) and the cyanobacterium ( Anacystis nidulans ) exhibited significant Ni adsorption in the order 91%, 72%, 75%, respectively, within 2 h contact with aqueous NiCl 2 (7·05 μg Ni/0·1 mg biomass). The immobilizing agent (Ca-alginate, 1·5%, w/v) absorbed more Ni (43%) than the exopolysaccharide of cyanobacteria, Rivularia sp. (40%) of Aphanothece sp. (30%). Ni remobilization from different adsorbed systems was maximum (84%) for Ca(NO 3 ) 2 over NaCl (4·3%) at equimolar concentrations (12 m M , each). Extracts from forest soil (organic C, 2–3%) were more effective in Ni remobilization (22·65%) than similar preparations from garden soil (18%) with organic C in the range of 0·98–1·1%.