Search results for: cyanobacterial engineering

Authors: Anurakti Shukla, Sudhakar Srivastava

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Cyanobacteria are known for rapid growth rates, high biomass, and the ability to accumulate potentially toxic elements and contaminants. The present work was planned to develop a low-cost, feasible prototype, CyanoClean, for the growth of a cyanobacterial consortium for the removal of arsenic (As) from water. The cyanobacterial consortium consisting of Oscillatoria, Phormidiumand Gloeotrichiawas used, and the conditions for optimal growth of the consortium were standardized. A pH of 7.6, initial cyanobacterial biomass of 10 g/L, and arsenite [As(III)] and arsenate [As(V)] concentration of 400 μΜand 600 μM, respectively, were found to be suitable. The CyanoClean prototype was designed with acrylic sheet and had arrangements for optimal cyanobacterial growth in natural sunlight and also in artificial light. The As removal experiments in concentration- and duration-dependent manner demonstrated removal of up to 39-69% and 9-33% As respectively from As(III) and As(V)-contaminated water. In field testing of CyanoClean, natural As-contaminated groundwater was used, and As reduction was monitored when a flow rate of 3 L/h was maintained. In a field experiment, As concentration in groundwater was found to reduce from 102.43 μg L⁻¹ to <10 μg L⁻¹ after 6 h in natural sunlight. However, in shaded conditions under artificial light, the same result was achieved after 9 h. The CyanoClean prototype is of simple design and can be easily up-scaled for application at a small- to medium-size land and shall be affordable even for a low- to middle-income group farmer.

Keywords: cyanoclean, gloeotrichia, oscillatoria, phormidium, phycoremediation

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Authors: Adam Bownik, Małgorzata Adamczuk, Barbara Pawlik-Skowrońska

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Certain strains of cyanobacteria, microorganisms forming water blooms, produce toxic secondary metabolites. Although various effects of cyanotoxins in aquatic animals are known, little data can be found on the influence of some cyanobacterial oligopeptides beyond microcystins. The aim of the present study was to determine the toxicity of novel pure cyanobacterial oligopeptides: microginin FR-1 (MGFR1) and anabaenopeptin-A (ANA-A) on a transparent model rotifer Brachionus calyciflorus with the use of fluorescent double staining with Hoechst 34580 and propidium iodide. The obtained results showed that both studied oligopeptides decreased the fluorescence intensity of animals stained with Hoechst 34580 in a concentration-dependent manner. On the other hand, a concentration-dependent increase of propidium iodide fluorescence was noted in the exposed rotifers. The results suggest that MGFR-1 and ANA-A should be considered as a potent toxic agent to freshwater rotifers, and fluorescent staining with Hoechst and propidium iodide may be a valuable tool for determination of toxicity of cyanobacterial oligopeptides in rotifers.

Keywords: cyanobacteria, brachionus, oligopeptides, fluorescent staining, hoechst, propidium iodide

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Authors: Kyou Hee Shim, Hwa Sung Shin

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When a thrombolysis agent is administered to treat ischemic stroke, excessive reactive oxygen species are generated due to a sudden provision of oxygen and occurs secondary damage cell necrosis. Thus, it is necessary to administrate adjuvant as well as thrombolysis agent to protect and reduce damaged tissue. As cerebral blood vessels have specific structure called blood-brain barrier (BBB), it is not easy to transfer substances from blood to tissue. Therefore, development of a drug carrier is required to increase drug delivery efficiency to brain tissue. In this study, cyanobacterial pigment from the blue-green algae known for having neuroprotective effect as well as antioxidant effect was nasally administrated for bypassing BBB. In order to deliver cyanobacterial pigment efficiently, the nano-sized liposome was used as a carrier. Liposomes were coated with a positive charge of chitosan since negative residues are present at the nasal mucosa the first gateway of nasal administration. Characteristics of liposome including morphology, size and zeta potential were analyzed by transmission electron microscope (TEM) and zeta analyzer. As a result of cytotoxic test, the liposomes were not harmful. Also, being administered a drug to the ischemic stroke animal model, we could confirm that the pharmacological effect of the pigment delivered by chitosan coated liposome was enhanced compared to that of non-coated liposome. Consequently, chitosan coated liposome could be considered as an optimized drug delivery system for the treatment of acute ischemic stroke.

Keywords: ischemic stroke, cyanobacterial pigment, liposome, chitosan, nasal administration

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Authors: Belal Bakheet, John Beardall, Xiwang Zhang, David McCarthy

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The potential risks associated with toxic cyanobacteria have raised growing environmental and public health concerns leading to an increasing effort into researching ways to bring about their removal from water, together with destruction of their associated cyanotoxins. A variety of toxins are synthesized by cyanobacteria and include hepatotoxins, neurotoxins, and cytotoxins which can cause a range of symptoms in humans from skin irritation to serious liver and nerve damage. Therefore drinking water treatment processes should ensure the consumers’ safety by removing both cyanobacterial cells, and cyanotoxins from the water. Cyanobacterial cells and cyanotoxins presented challenges to the conventional water treatment systems; their accumulation within drinking water treatment plants has been reported leading to plants shut down. Thus, innovative and effective water purification systems to tackle cyanobacterial pollution are required. In recent years there has been increasing attention to the electrochemical oxidation process as a feasible alternative disinfection method which is able to generate in situ a variety of oxidants that would achieve synergistic effects in the water disinfection process and toxin degradation. By utilizing only electric current, the electrochemical process through electrolysis can produce reactive oxygen species such as hydroxyl radicals from the water, or other oxidants such as chlorine from chloride ions present in the water. From extensive physiological and morphological investigation of cyanobacterial cells during electrolysis, our results show that these oxidants have significant impact on cell inactivation, simultaneously with cyanotoxins removal without the need for chemicals addition. Our research aimed to optimize existing electrochemical oxidation systems and develop new systems to treat water containing toxic cyanobacteria and cyanotoxins. The research covers detailed mechanism study on oxidants production and cell inactivation in the treatment under environmental conditions. Overall, our study suggests that the electrochemical treatment process e is an effective method for removal of toxic cyanobacteria and cyanotoxins.

Keywords: toxic cyanobacteria, cyanotoxins, electrochemical process, oxidants

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Authors: Vandana Deopanée Tomar, Gurpreet Kaur Sidhu, Panchsheela Nogia, Rajesh Mehrotra, Sandhya Mehrotra

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The cyanobacterial CO₂ concentrating mechanism (CCM) operates to raise the levels of CO₂ in the vicinity of the main carboxylation enzyme Rubisco which is encapsulated in protein micro compartments called carboxysomes. Thus, due to the presence of CCM, cyanobacterial cells are able to work with high photosynthetic efficiency even at low Ci conditions and can accumulate 1000 folds high internal concentrations of Ci than external environment. Engineering of some useful CCM components into higher plants is one of the plausible approaches to improve their photosynthetic performance. The first step and the simplest approach for attaining this objective would be the transfer of cyanobacterial bicarbonate transporter such as SbtA to inner chloroplast envelope of C₃ plants. For this, SbtA transporter gene from Synechococcus elongatus PCC 7942 was fused to a transit peptide element to generate chimeric constructs in order to direct it to chloroplast inner envelope. Two transit peptides namely, TnaXTP (transit peptide from AT3G56160) and TMDTP (transit peptide from AT2G02590) were shortlisted from Arabidopsis thaliana genome and cloned in plant expression vector pCAMBIA1302 having mgfp5 as a reporter gene. Plant transformation was done by agro infiltration and Agrobacterium mediated co-culture. DNA, RNA, and protein were isolated from the leaves four days post infiltration, and the presence of transgene was confirmed by gene specific PCR (Polymerase Chain Reaction) analysis and by RT-PCR (Reverse Transcription Polymerase Chain Reaction). The expression was confirmed at the protein level by western blotting using anti-GFP primary antibody and horseradish peroxidase (HRP) conjugated secondary antibody. The localization of the protein was detected by confocal microscopy of isolated protoplasts. We observed chloroplastic expression for both the fusion constructs which suggest that the transit peptide sequences are capable of taking the cargo protein to the chloroplasts. These constructs are now being used to generate stable transgenic plants by Agrobacterium mediated transformation. The stability of transgene expression will be analyzed from T₀ to T₂ generation.

Keywords: agro infiltration, bicarbonate transporter, carbon concentrating mechanisms, cyanobacteria, SbtA

Procedia PDF Downloads 184

Authors: Sujatha Edla

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Introduction: New compounds and possible uses for the bioactive substances produced by freshwater cyanobacteria are constantly being discovered through research. Certain molecules are hazardous to the environment and human health, but others have potential applications in industry, biotechnology, and pharmaceuticals. These discoveries advance our knowledge of the varied functions these microbes perform in different ecosystems. Cyanobacterial silver nanoparticles (AgNPs) have special qualities and possible therapeutic advantages, which make them very promising for a range of medicinal uses. Aim: In our study; the attention was focused on the analysis and characterization of bioactive compounds extracted from freshwater cyanobacteria Planktothricoides raciorskii and its comparative study on Cyanobacteria-mediated silver nanoparticles synthesized by cell-free extract of Planktothricoides raciorskii. Material and Methods: A variety of bioactive secondary metabolites have been extracted, purified, and identified from cyanobacterial species using column chromatography, FTIR, and GC-MS/MS chromatography techniques and evaluated for antibacterial and cytotoxic studies, where the Cyanobacterial silver nanoparticles (CSNPs) were characterized by UV-Vis spectroscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM), and Fourier transform infrared (FTIR) analysis and were further tested for antibacterial and cytotoxic efficiency. Results: The synthesis of CSNPs was confirmed through visible color change and shift of peaks at 430–445 nm by UV-Vis spectroscopy. The size of CSNPs was between 22 and 34 nm and oval-shaped which were confirmed by SEM and TEM analyses. The FTIR spectra showed a new peak at the range of 3,400–3,460 cm−1 compared to the control, confirming the reduction of silver nitrate. The antibacterial activity of both crude bioactive compound extract and CSNPs showed remarkable activity with Zone of inhibition against E. coli with 9.5mm and 10.2mm, 13mm and 14.5mm against S. paratyphi, 9.2mm and 9.8mm zone of inhibition against K. pneumonia by both crude extract and CSNPs, respectively. The cytotoxicity as evaluated by extracts of Planktothricoides raciorskii against MCF7-Human Breast Adenocarcinoma cell line and HepG2- Human Hepatocellular Carcinoma cell line employing MTT assay gave IC50 value of 47.18ug/ml, 110.81ug/ml against MCF7cell line and HepG2 cell line, respectively. The cytotoxic evaluation of Planktothricoides raciorskii CSNPs against the MCF7cell line was 43.37 ug/ml and 20.88 ug/ml against the HepG2 cell line. Our ongoing research in this field aims to uncover the full therapeutic potential of cyanobacterial silver nanoparticles and address any associated challenges.

Keywords: cyanobacteria, silvernanoparticles, pharmaceuticals, bioactive compounds, cytotoxic

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Authors: K. Thiel, P. Patrikainen, C. Nagy, D. Fitzpatrick, E.-M. Aro, P. Kallio

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Photosynthetic cyanobacteria have been recognized as potential future biotechnological hosts for the direct conversion of CO₂ into chemicals of interest using sunlight as the solar energy source. However, in order to develop commercially viable systems, the flux of electrons from the photosynthetic light reactions towards specified target chemicals must be significantly improved. The objective of the study was to investigate whether the autotrophic production efficiency of specified end-metabolites can be improved in engineered cyanobacterial cells by rescuing excited electrons that are normally lost to molecular oxygen due to the cyanobacterial flavodiiron protein Flv1/3. Natively Flv1/3 dissipates excess electrons in the photosynthetic electron transfer chain by directing them to molecular oxygen in Mehler-like reaction to protect photosystem I. To evaluate the effect of flavodiiron inactivation on autotrophic production efficiency in the cyanobacterial host Synechocystis sp. PCC 6803 (Synechocystis), sucrose was selected as the quantitative reporter and a representative of a potential end-product of interest. The concept is based on the native property of Synechocystis to produce sucrose as an intracellular osmoprotectant when exposed to high external ion concentrations, in combination with the introduction of a heterologous sucrose permease (CscB from Escherichia coli), which transports the sucrose out from the cell. In addition, cell growth, photosynthetic gas fluxes using membrane inlet mass spectrometry and endogenous storage compounds were analysed to illustrate the consequent effects of flv deletion on pathway flux distributions. The results indicate that a significant proportion of the electrons can be lost to molecular oxygen via Flv1/3 even when the cells are grown under high CO₂ and that the inactivation of flavodiiron activity can enhance the photosynthetic electron flux towards optionally available sinks. The flux distribution is dependent on the light conditions and the genetic context of the Δflv mutants, and favors the production of either sucrose or one of the two storage compounds, glycogen or polyhydroxybutyrate. As a conclusion, elimination of the native Flv1/3 reaction and concomitant introduction of an engineered product pathway as an alternative sink for excited electrons could enhance the photosynthetic electron flux towards the target endproduct without compromising the fitness of the host.

Keywords: cyanobacterial engineering, flavodiiron proteins, redirecting electron flux, sucrose

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Authors: Zuzana Sedrlova, Eva Slaninova, Ines Fritz, Christina Daffert, Stanislav Obruca

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Cyanobacteria are ecologically extremely important phototrophic gram-negative bacteria capable of oxygenic photosynthesis. They synthesize many interesting metabolites such as glycogen, carotenoids, but the most interesting metabolites are polyhydroxyalkanoates (PHA). The main advantage of cyanobacteria is the fact they do not require costly organic substrate and, oppositely, cyanobacteria can fix CO₂. PHA serves primarily as a carbon and energy source and occurs in the form of intracellular granules in bacterial cells. It is possible, PHA helps cyanobacteria to survive stress conditions since increased PHA synthesis was observed during cultivation in stress conditions. PHA is microbial biopolymers that are biodegradable with similar properties as petrochemical synthetic plastics. Production of PHA by heterotrophic bacteria is expensive; for price reduction waste materials as input, materials are used. Positively, cyanobacteria principally do not require organic carbon substrate since they are capable of CO₂ fixation. In this work, we demonstrated that stress conditions lead to the highest obtained yields of PHA in cyanobacterial cultures. Two cyanobacterial cultures from genera Synechocystis were used in this work. Cultivations were performed either in Erlenmayer flask or in tube multicultivator. Multiple stressors were applied on cyanobacterial cultures, and stressors include PHA precursors. PHA precursors are chemical substances and some of them do not occur naturally in the environment. Cultivation with the same PHA precursors in the same concentration led to a 1,6x higher amount of PHA when a multicultivator was used. The highest amount of PHA reached 25 % of PHA in dry cyanobacterial biomass. Both strains are capable of co-polymer synthesis in the presence of their structural precursor. The composition of co-polymer differs in Synechocystis sp. PCC 6803 and Synechocystis salina CCALA 192. Synechocystis sp. PCC 6803 cultivated with γ-butyrolakton accumulated co-polymer of 3-hydroxybutyrate (3HB) and 4-hydroxybutyrate (4HB) the composition of the copolymer was 56 % of 4HB and 44 % of 3HB. The total amount of PHA, as well as yield of biomass, was lower than in control due to the toxic properties of γ-butyrolakton. Funding: This study was partly funded by the project GA19- 19-29651L of the Czech Science Foundation (GACR) and partly funded by the Austrian Science Fund (FWF), a project I 4082-B25. This work was supported by Brno, Ph.D. Talent – Funded by the Brno City Municipality.

Keywords: co-polymer, cyanobacteria, PHA, synechocystis

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Authors: Shivam Yadav, Neelam Atri

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Cyanobacteria are photoautotrophic prokaryotes able to grow in diverse ecological habitats, originated 2.5 - 3.5 billion years ago and brought oxygenic photosynthesis. Since then superoxide dismutases (SODs) acquired great significance due to their ability to catalyze detoxification of byproducts of oxygenic photosynthesis, i.e. superoxide radicals. Sequence information from several cyanobacterial genomes offers a unique opportunity to conduct a comprehensive comparative analysis of the superoxide dismutases family. In the present study, we extracted information regarding SODs from species of sequenced cyanobacteria and investigated their diversity, conservation, domain structure, and evolution. 144 putative SOD homologues were identified. SODs are present in all cyanobacterial species reflecting their significant role in survival. However, their distribution varies, fewer in unicellular marine strains whereas abundant in filamentous nitrogen-fixing cyanobacteria. Motifs and invariant amino acids typical in eukaryotic SODs were conserved well in these proteins. These SODs were classified into three major families according to their domain structures. Interestingly, they lack additional domains as found in proteins of other family. Phylogenetic relationships correspond well with phylogenies based on 16S rRNA and clustering occurs on the basis of structural characteristics such as domain organization. Similar conserved motifs and amino acids indicate that cyanobacterial SODs make use of a similar catalytic mechanism as eukaryotic SODs. Gene gain-and-loss is insignificant during SOD evolution as evidenced by absence of additional domain. This study has not only examined an overall background of sequence-structure-function interactions for the SOD gene family but also revealed variation among SOD distribution based on ecophysiological and morphological characters.

Keywords: comparative genomics, cyanobacteria, phylogeny, superoxide dismutases

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Authors: Shivam Yadav, Neelam Atri

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Cyanobacteria, the photosynthetic prokaryotes are indispensable components of paddy soil contribute substantially to the nitrogen economy however often appended with metal load. They are well known to play crucial roles in maintenance of soil fertility and rice productivity. Nickel is one such metal that plays a vital role in the cellular physiology, however at higher concentrations it exerts adverse effects. Arsenic is another toxic metalloid that negatively affects the cyanobacterial proliferation. However species-specific comparative responses under As and Ni is largely unknown. The present study focuses on the comparative effects of nickel (Ni2+) and arsenite (As(III)) on two diazotrophic cyanobacterial species (Anabaena doliolum and Anabaena sp. PCC7120) in terms of antioxidative aspects. Oxidative damage measured in terms of lipid peroxidation and peroxide content was significantly higher after As(III) than Ni treatment as compared to control. Similarly, all the studied enzymatic and non-enzymatic parameters of antioxidative defense system except glutathione reductase (GR) showed greater induction against As(III) than Ni. Moreover, integrating comparative analysis of all studied parameters also demonstrated interspecies variation in terms of stress adaptive strategies reflected through higher sensitivity of Anabaena doliolum over Anabaena PCC7120.

Keywords: antioxidative system, arsenic, cyanobacteria, nickel

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Authors: Moletsane Makgotso, Mogakabe Elijah, Marrengane Zinhle

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South Africa’s water resources quality is becoming more and more weakened by eutrophication, which deteriorates its usability. Thirty five percent of fresh water resources are eutrophic to hypertrophic, including grossly-enriched reservoirs that go beyond the globally-accepted definition of hypertrophy. Failing infrastructure adds to the problem of contaminated urban runoff which encompasses an important fraction of flows to inland reservoirs, particularly in the non-coastal, economic heartland of the country. Eutrophication threatens the provision of potable and irrigation water in the country because of the dependence on fresh water resources. Eutrophicated water reservoirs increase water treatment costs, leads to unsuitability for recreational purposes and health risks to human and animal livelihood due to algal proliferation. Eutrophication is caused by high concentrations of phosphorus and nitrogen in water bodies. In South Africa, Microsystis and Anabaena are widely distributed cyanobacteria, with Microcystis being the most dominant bloom-forming cyanobacterial species associated with toxin production. Two impoundments were selected, namely the Klipvoor and Leeukraal dams as they are mainly used for fishing, recreational, agricultural and to some extent, potable water purposes. The total oxidized nitrogen and total phosphorus concentration were determined as causative nutrients for eutrophication. Chlorophyll a and total microcystins, as well as the identification of cyanobacteria was conducted as indicators of cyanobacterial infestation. The orthophosphate concentration was determined by subjecting the samples to digestion and filtration followed by spectrophotometric analysis of total phosphates and dissolved phosphates using Aquakem kits. The total oxidized nitrates analysis was conducted by initially conducting filtration followed by spectrophotometric analysis. Chlorophyll a was quantified spectrophotometrically by measuring the absorbance of before and after acidification. Microcystins were detected using the Quantiplate Microcystin Kit, as well as microscopic identification of cyanobacterial species. The Klipvoor dam was found to be hypertrophic throughout the study period as the mean Chlorophyll a concentration was 269.4µg/l which exceeds the mean value for the hypertrophic state. The mean Total Phosphorus concentration was >0.130mg/l, and the total microcystin concentration was > 2.5µg/l throughout the study. The most predominant algal species were found to be the Microcystis. The Leeukraal dam was found to be mesotrophic with the potential of it becoming eutrophic as the mean concentration for chlorophyll a was 18.49 µg/l with the mean Total Phosphorus > 0.130mg/l and the Total Microcystin concentration < 0.16µg/l. The cyanobacterial species identified in Leeukraal have been classified as those that do not pose a potential risk to any impoundment. Microcystis was present throughout the sampling period and dominant during the warmer seasons. The high nutrient concentrations led to the dominance of Microcystis that resulted in high levels of microcystins rendering the impoundments, particularly Klipvoor undesirable for utilisation.

Keywords: nitrogen, phosphorus, cyanobacteria, microcystins

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Authors: Ravi Raghav Sonani, Niraj Kumar Singh, Jitendra Kumar, Datta Madamwar

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Phycobiliproteins (PBPs) are light harvesting proteins showing very strong absorbance and fluorescence in the visible range of the solar spectrum. Phycoerythrin (PE) and phycocyanin (PC) are majorly found PBPs in the cyanobacteria and red algae. In the present study, we have investigated the reactive oxygen species (ROS)-averting capacity of purified PE and PC of cyanobacterial origin. Furthermore, the possibility - whether the ROS-averting potential of PBPs can be explored in the therapeutics of oxidative stress associated physiological anomalies including aging and neurodegenerative diseases. The nematode Caenorhabditis elegans has been used as model organism in this study. PE and PC treatment moderated normal aging and associated physiological functionalities like pharyngeal pumping and locomotion of C. elegans. Moreover, PE-treatment enhanced the stress (oxidative and heat) tolerance upon PE and PC treatment. Specifically, PE treatment was also noted to moderate the progression of Alzheimer’s disease in transgenic C. elegans CL4176. However, PC-treatment curtailed the polyQ aggregation mediated proteotoxicity in C. elegans AM141 (Huntington disease model) under stressed (paraquat stress) as well as normal conditions. The effectiveness of PE and PC in expanding the lifespan of mutant C. elegans knockout for some up- (daf 16) and down- (daf-2 and age-1) stream regulators of insulin/IGF-1 signalling (IIS) shows the independency of their effects from DAF-2–AGE-1–DAF-16 signalling pathway. In conclusion, the present report demonstrates the anti-aging and neuro-protective potential of cyanobacterial PE and PC.

Keywords: phycobiliproteins, aging, alzheimer, huntington, C. elegans

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Authors: Laura Valdés-Santiago, José Luis Castro-Guillén, Jorge Noé García-Chávez, Rosalba Alonso-Rodríguez, Rafael Vargas-Bernal

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The quality and availability of the water contained in dams (artificial bodies of water) are at risk due to the presence of uncontrolled growths of cyanobacteria capable of producing cyanotoxins that affect the ecosystem and harm the health of humans and animals. The physicochemical properties were measured, and the degree of eutrophy of four dams from Guanajuato was determined. They presented a pH of 6.1 to 8.4, conductivity of 121 to 415 μS/cm², chlorophyll of 0.43-42.43 μg/L, NO₃- 0-1.2 mg/L and PO₄3- 0.11 to 0.84 mg/L; considering these parameters, the prey most prone to the development of cyanobacterial blooms were El Palote dam, La Purísima dam, and Allende dam, but not El Conejo dam. The potentially toxic cyanobacteria identified were Planktothrix agardhii, Oscillatoria sp., Raphidiopsis sp., and Microcystis sp., Microcystin-LR, Nodularin, and Cylindrospermopsin were quantified, presenting values between 0.08-0.42 and 0.02-2.05 ppb, respectively, the water bodies with the highest concentration were El Palote dam and La Purísima dam. Microcystin-LR and/or Nodularin levels are within the guideline values for human consumption in drinking water established by the World Health Organization for Microcystin-LR and for Cylindrospermopsin by the Oregon Health Authority (OHA) in all dams. This work is relevant due to the use of these bodies of water for agriculture and human consumption in the state, and the presence of toxin-producing cyanobacteria can represent an environmental, ecotoxicological, and health problem, so it is recommended to establish a program of frequent monitoring of cyanobacteria and cyanotoxins in the state's dams.

Keywords: Planktrothrix agardhii, Raphidiopsis sp., Microcystis sp., Cyanobacterial blooms, Cyanotoxins

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Authors: Medina Kadiri

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Blue-green otherwise called cyanobacteria, produce an array of biotoxins grouped into five categories notably hapatotoxins, neurotoxins, cytotoxins, dermatotoxins, and irritant toxins. Microcystins which are examples of hepatotoxins produced by blue-green algae Microcystins comprise the most common group of the cyanobacterial toxins. Blue-green algae flourish in aquatic environments, whether marine, brackish or freshwater, producing blooms in different forms such as microscopic, mats, or unsightly odoriferous scums. Microcystins biotoxins cause a plethora of animal and human hazards such as liver damage/cirrhosis and cancer, kidney damage, dermatitis, tinnitus, gastroenteritis, sore throat, nausea, myalgia, neurological problems, respiratory irritation and death. Water samples were collected from coastal regions of Nigeria in March 2014, June 2014, October 2014 and January 2015 and analyzed with Enzyme Linked Immunosorbent Assay (ELISA) kits. Microcystin biotoxin was recorded in all sites both during dry and wet seasons. The range of microcystins found was 0.000041-There was a seasonal trend of increasing microcystin concentrations from March till Octobers and a decrease thereafter. Generally in the oceanic waters, microcystin levels were highest at Cross Rivers in March and January, Barbeach in June and Lekki in October. In the adjoining riverine ecosystems, on the other hand, the highest concentrations of microcystin were observed at Akwa Ibom in March, June and October and in Bayelsa in January. Continuous monitoring and screening of coastal water bodies is suggested to minimize the health risks of cyanobacterial biotoxins to coastal communities of Nigeria.

Keywords: biotoxins, harmful algae, marine, microcystin, Nigeria

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Authors: J. M. Kim, Y. W. Jung, E. Lee, Y. K. Kwack, , S. K. Sim*

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Cyanobacterial blooms in lakes, reservoirs, and rivers have been environmental and social issues due to its toxicity, odor, etc. Among the cyanotoxins, microcystins exist mostly within the cyanobacterial cells, and they are released from the cells. Therefore, an innovative technology is needed to detoxify the harvested microalgae for environment-friendly utilization of the harvested microalgae. This study develops detoxification method of microcystins in the harvested microalgae and recycling harvested microalgae with food waste using salt-tolerant mushroom strains and natural ecosystem decomposer. During this eco-friendly organic waste recycling process, diverse bacteria or various enzymes of the salt-tolerant mushroom strains decompose the microystins and cyclic peptides. Using PHLC/Mass analysis, it was verified that 99.8% of the microcystins of the harvested microalgae was detoxified in the harvested mushroom as well as in the recycled organic biomass. Further study is planned to verify the decomposition mechanisms of the microcystins by the bacteria or enzymes. In this study, the harvested microalgae is mixed with the food waste, and then the mixed toxic organic waste is used as mushroom compost by adjusting the water content of about 70% using cellulose such as sawdust cocopeats and cottonseeds. The mushroom compost is bottled, sterilized, and salt-tolerant mushroom spawn is inoculated. The mushroom is then cultured and growing in the temperature, humidity, and CO2 controlled environment. During the cultivation and growing process of the mushroom, microcystins are decomposed into non-toxic organic or inorganic compounds by diverse bacteria or various enzymes of the mushroom strains. Various enzymes of the mushroom strains decompose organics of the mixed organic waste and produce nutritious and antibiotic mushrooms. Cultured biomass compost after mushroom harvest can be used for organic fertilizer, functional bio-feed, and RE-100 biomass renewable energy source. In this eco-friendly organic waste recycling process, no toxic material, wastewater, nor sludge is generated; thus, sustainable with the circular economy.

Keywords: microalgae, microcystin, food waste, salt-tolerant mushroom strains, sustainability, circular economy

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Authors: Krishna N. Gaikwad, V. R. Kakulte

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Rice (Oryza sativa L.) is the main cereal crop of tribal people of western part of Nasik district. There is a wide fluctuation in yield due to the factors like uncertain rains, pest diseases, socio-economic status of farmers, lack of awareness and traditional knowledge of farmers about agro-practices. In order to achieve more yield, it is a need to adopt low cost, eco-friendly blue green algal biofertilizer technology. Communication of useful information to needy people is basic need in present situation. The paper reports different communication modes of paddy technologies, adoption about BGA technology, attitudinal changes of farmers and yield of rice production during year 2011 and 2012. The results indicate that there is significant effect of communication modes of improved BGA technology on rice yield.

Keywords: rice, BGA, biofertilizer, Oryza sativa L.

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Authors: J. Pazourek, K. Šmejkal, P. Kollár, J. Rajchard, J. Šinko, Z. Balounová, E. Vlková, H. Salmonová

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Pectinatella magnifica (Leidy, 1851) is an invasive freshwater animal that lives in colonies. A colony of Pectinatella magnifica (a gelatinous blob) can be up to several feet in diameter large and under favorable conditions it exhibits an extreme growth rate. Recently European countries around rivers of Elbe, Oder, Danube, Rhine and Vltava have confirmed invasion of Pectinatella magnifica, including freshwater reservoirs in South Bohemia (Czech Republic). Our project (Czech Science Foundation, GAČR P503/12/0337) is focused onto biology and chemistry of Pectinatella magnifica. We monitor the organism occurrence in selected South Bohemia ponds and sandpits during the last years, collecting information about physical properties of surrounding water, and sampling the colonies for various analyses (classification, maps of secondary metabolites, toxicity tests). Because the gelatinous matrix is during the colony lifetime also a host for algae, bacteria and cyanobacteria (co-habitants), in this contribution, we also applied a high performance liquid chromatography (HPLC) method for determination of potentially present cyanobacterial toxins (microcystin-LR, microcystin-RR, nodularin). Results from the last 3-year monitoring show that these toxins are under limit of detection (LOD), so that they do not represent a danger yet. The final goal of our study is to assess toxicity risks related to fresh water resources invaded by Pectinatella magnifica, and to understand the process of invasion, which can enable to control it.

Keywords: cyanobacteria, fresh water resources, Pectinatella magnifica invasion, toxicity monitoring

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Authors: Ruchi Pathania, Ahmad Ahmad, Shireesh Srivastava

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Cyanobacteria is a promising microbe that can capture and convert atmospheric CO₂ and light into valuable industrial bio-products like biofuels, biodegradable plastics, etc. Among their most attractive traits are faster autotrophic growth, whole year cultivation using non-arable land, high photosynthetic activity, much greater biomass and productivity and easy for genetic manipulations. Cyanobacteria store carbon in the form of glycogen which can be hydrolyzed to release glucose and fermented to form bioethanol or other valuable products. Marine cyanobacterial species are especially attractive for countries with scarcity of freshwater. We recently identified a marine native cyanobacterium Synechococcus sp. BDU 130192 which has good growth rate and high level of polyglucans accumulation compared to Synechococcus PCC 7002. In this study, firstly we sequenced the whole genome and the sequences were annotated using the RAST server. Genome scale metabolic model (GSMM) was reconstructed through COBRA toolbox. GSMM is a computational representation of the metabolic reactions and metabolites of the target strain. GSMMs construction through the application of Flux Balance Analysis (FBA), which uses external nutrient uptake rates and estimate steady state intracellular and extracellular reaction fluxes, including maximization of cell growth. The model, which we have named isyn942, includes 942 reactions and 913 metabolites having 831 metabolic, 78 transport and 33 exchange reactions. The phylogenetic tree obtained by BLAST search revealed that the strain was a close relative of Synechococcus PCC 7002. The flux balance analysis (FBA) was applied on the model iSyn942 to predict the theoretical yields (mol product produced/mol CO₂ consumed) for native and non-native products like acetone, butanol, etc. under phototrophic condition by applying metabolic engineering strategies. The reported strain can be a viable strain for biotechnological applications, and the model will be helpful to researchers interested in understanding the metabolism as well as to design metabolic engineering strategies for enhanced production of various bioproducts.

Keywords: cyanobacteria, flux balance analysis, genome scale metabolic model, metabolic engineering

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Authors: Nasim Rasuli, Akram Ahmadi, Hossein Riahi, Zeinab Shariatmadari, Majid Ghorbani Nohooji, Pooyan Mehraban Joubani

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Cyanobacteria are one of the most promising sources of new biostimulants and have received much attention due to their diverse applications in biotechnology. These microorganisms enhance the growth and productivity of plants by producing plant growth stimulants and fixing atmospheric nitrogen. Thymus vulgaris L., a valuable medicinal plant from the Lamiaceae family, is widely distributed across the globe. essential oil of T. vulgaris is best characterized by the prominence of phenols, making them the key compounds in its composition. Lignin biosynthesis as a natural plant polyphenol plays a crucial role in promoting plant growth, strengthening cell walls, and increasing resistance to pathogens. In this study, the bioelicitor activity of five cyanobacterial suspensions including Anabaena torulosa ISB213, Nostoc calcicola ISB215, Nostoc ellipsosporum ISB217, Trichormus doliolum ISB214, and Oscillatoria sp. ISB2116 on the lignin content of the T. vulgaris L. was investigated. Pot experiments were performed by inoculation of a %2 algal extract to the soil of treated plants one week before planting and then every 20 days. After four months, the lignin content in the leaves of both treated and control plants was evaluated. The results demonstrated that the application of cyanobacteria significantly increased the lignin content in the leaves of treated plants compared to the control. The treatment with Oscillatoria sp. ISB216 and N. ellipsosporum ISB217 resulted in the highest lignin content, with an increase of 93.33% and 86.67%, respectively. These findings highlight the potential of cyanobacteria as bioelicitors, offering a viable alternative for enhancing the production of secondary metabolites in T. vulgaris. Consequently, this could contribute to the economic value of this medicinal plant.

Keywords: cyanobacteria, bioelicitor, thymus vulgaris, lignin

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Authors: Julie Connelly, Tanvi Toprani, Xin Xie, Dhinoth Kumar Bangarusamy, Kris C. Gunsalus

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The overuse of antibiotics worldwide has contributed to the development of multi-drug resistant (MDR) pathogenic bacterial strains. There is an increasing urgency to discover antibiotics to combat MDR pathogens. The microbiome of the Arabian Gulf is a largely unexplored and potentially rich source of novel bioactive compounds. Microbes that inhabit the Abu Dhabi coastal regions adapt to extreme environments with high salinity, hot temperatures, large temperature fluctuations, and acute exposure to solar energy. The microbes native to this region may produce unique metabolites with therapeutic potential as antibiotics and antifungals. We have isolated 200 pure bacterial strains from mangrove sediments, cyanobacterial mats, and coral reefs of the Abu Dhabi region. In this project, we aim to screen the marine bacterial strains to identify antibiotics, in particular undocumented compounds that show activity against existing antibiotic-resistant strains. We have acquired the ESKAPE pathogen panel, which consists of six antibiotic-resistant gram-positive and gram-negative bacterial pathogens that collectively cause most clinical infections. Our initial efforts of the primary screen using colony-picking co-culture assay have identified several candidate marine strains producing potential antibiotic compounds. We will next apply different assays, including disk-diffusion and broth turbidity growth assay, to confirm the results. This will be followed by bioactivity-guided purification and characterization of target compounds from the scaled-up volume of candidate strains, including SPE fraction, HPLC fraction, LC-MS, and NMR. For antimicrobial compounds with unknown structures, our final goal is to investigate their mode of action by identifying the molecular target.

Keywords: marine bacteria, natural products, drug discovery, ESKAPE panel

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Authors: Nilesh Bandekar, Sumita Dasgupta, Luis Alberto Allcahuaman Huaya, Souvik Manna

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Cryptobiosis is a dormant state where all measurable metabolic activities are at a halt, allowing an organism to survive in extreme conditions like low temperature (cryobiosis), extreme drought (anhydrobiosis), etc. This phenomenon is observed especially in tardigrades that can retain this state for decades depending on the abiotic environmental conditions. On returning to favorable conditions, tardigrades re-attain a metabolically active state. In this study, cyanobacteria as a model organism are being chosen to induce cryptobiosis for its effective preservation over a long period of time. Preserving cyanobacteria using this strategy will have multiple space applications because of its ability to produce oxygen. In addition, research has shown the survivability of this organism in space for a certain period of time. Few species of cyanobacterial residents of the soil such as Microcoleus, are able to survive in extreme drought as well. This work specifically focuses on Synechococcus elongatus, an endolith cyanobacteria with multiple benefits. It has the capability to produce 25% oxygen in water bodies. It utilizes carbon dioxide to produce oxygen via photosynthesis and also uses carbon dioxide as an energy source to form glucose via the Calvin cycle. There is a fair possibility of initiating cryptobiosis in such an organism by inducing certain proteins extracted from tardigrades such as Heat Shock Proteins (Hsp27 and Hsp30c) and/or hydrophilic Late Embryogenesis Abundant proteins (LEA). Existing methods like cryopreservation are difficult to execute in space keeping in mind their cost and heavy instrumentation. Also, extensive freezing may cause cellular damage. Therefore, cryptobiosis-induced cyanobacteria for its transportation from Earth to Mars as a part of future terraforming missions on Mars will save resources and increase the effectiveness of preservation. Finally, Cyanobacteria species like Synechococcus elongatus can also produce oxygen and glucose on Mars in favorable conditions and holds the key to terraforming Mars.

Keywords: cryptobiosis, cyanobacteria, glucose, mars, Synechococcus elongatus, tardigrades

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Authors: Simona Lucakova, Irena Branyikova

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Eutrophication, a condition when water becomes over-enriched with nutrients (P, N), can lead to undesirable excessive growth of phytoplankton, so-called algal bloom. This process results in the accumulation of toxin-producing cyanobacteria and oxygen depletion, both possibly leading to the collapse of the whole ecosystem. In real conditions, the limiting nutrient, which determines the possible growth of harmful algal bloom, is usually phosphorus. Algicides or flocculants have been applied in the eutrophicated waterbody in order to reduce the phytoplankton growth, which leads to the introduction of toxic chemicals into the water. In our laboratory, the idea of the prevention of harmful phytoplankton growth by the intentional cultivation of non-toxic cyanobacteria Tolypothrix tenuis in semi-open floating photobioreactors directly on the surface of phosphorus-rich waterbody is examined. During the process of cultivation, redundant phosphorus is incorporated into cyanobacterial biomass, which can be subsequently used for the production of biofuels, cosmetics, pharmaceuticals, or biostimulants for agricultural use. To determine the ability of phosphorus incorporation, batch-cultivation of Tolypothrix biomass in media simulating eutrophic water (10% BG medium) and in effluent from municipal wastewater treatment plant, both with the initial phosphorus concentration in the range 0.5-1.0 mgP/L was performed in laboratory-scale models of floating photobioreactors. After few hours of cultivation, the phosphorus content was decreased below the target limit of 0.035 mgP/L, which was given as a borderline for the algal bloom formation. Under laboratory conditions, the effect of several parameters on the rate of phosphorus decrease was tested (illumination, temperature, stirring speed/aeration gas flow, biomass to medium ratio). Based on the obtained results, a bench-scale floating photobioreactor was designed and will be tested for Tolypothrix growth in real conditions. It was proved that intentional cultivation of cyanobacteria Tolypothrix could be a suitable approach for extracting redundant phosphorus from eutrophic waters as prevention of algal bloom formation.

Keywords: cyanobacteria, eutrophication, floating photobioreactor, Tolypothrix

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Authors: Eva Slaninova, Diana Cernayova, Zuzana Sedrlova, Katerina Mrazova, Petr Sedlacek, Jana Nebesarova, Stanislav Obruca

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Cyanobacteria are gram-negative prokaryotes belonging to a group of photosynthetic bacteria. In comparison with heterotrophic microorganisms, cyanobacteria utilize atmospheric nitrogen and carbon dioxide without any additional substrates. This ability of these microorganisms could be employed in biotechnology for the production of bioplastics, concretely polyhydroxyalkanoates (PHAs) which are primarily accumulated as a storage material in cells in the form of intracellular granules. In this study, there two cyanobacterial cultures from genera Synechocystis were used, namely Synechocystic sp. PCC 6803 and Synechocystis salina CCALA 192. There were optimized and used several various approaches, including microscopic techniques such as cryo-scanning electron microscopy (Cryo-SEM) and transmission electron microscopy (TEM), and fluorescence lifetime imaging microscopy using Nile red as a fluorescent probe (FLIM). Due to these instrumental techniques, the morphology of intracellular space and surface of cells were characterized. The next group of methods which were employed was spectroscopic techniques such as UV-Vis spectroscopy measured in two modes (turbidimetry and integration sphere) and Fourier transform infrared spectroscopy (FTIR). All these diverse techniques were used for the detection and characterization of pigments (chlorophylls, carotenoids, phycocyanin, etc.) and PHAs, in our case poly (3-hydroxybutyrate) (P3HB). To verify results, gas chromatography (GC) was employed concretely for the determination of the amount of P3HB in biomass. Cyanobacteria were also characterized as polyhydroxybutyrate producers by flow cytometer, which could count cells and at the same time distinguish cells including P3HB and without due to fluorescent probe called BODIPY and live/dead fluorescent probe SYTO Blue. Based on results, P3HB content in cyanobacteria cells was determined, as also the overall fitness of the cells. Acknowledgment: Funding: This study was partly funded by the projectGA19-29651L of the Czech Science Foundation (GACR) and partly funded by the Austrian Science Fund (FWF), project I 4082-B25.

Keywords: cyanobacteria, fluorescent probe, microscopic techniques, poly(3hydroxybutyrate), spectroscopy, chromatography

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Authors: Apurva Gupta, Surendra Singh

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Spirulina spp., as a promising source of many commercially valuable products, is grown photo autotrophically in open ponds and raceways on a large scale. However, the economic exploitation in an open system seems to have been limited because of lack of multiple stress-tolerant strains. The present study aims to isolate a stable stress tolerant mutant of Spirulina platensis with improved growth rate and enhanced potential to produce its commercially valuable bioactive compounds. N-methyl-n'-nitro-n-nitrosoguanidine (NTG) at 250 μg/mL (concentration permitted 1% survival) was employed for chemical mutagenesis to generate random mutants and screened against NaCl. In a preliminary experiment, wild type S. platensis was treated with NaCl concentrations from 0.5-1.5 M to calculate its LC₅₀. Mutagenized colonies were then screened for tolerance at 0.8 M NaCl (LC₅₀), and the surviving colonies were designated as NaCl tolerant mutants of S. platensis. The mutant cells exhibited 1.5 times improved growth against NaCl stress as compared to the wild type strain in control conditions. This might be due to the ability of the mutant cells to protect its metabolic machinery against inhibitory effects of salt stress. Salt stress is known to adversely affect the rate of photosynthesis in cyanobacteria by causing degradation of the pigments. Interestingly, the mutant cells were able to protect its photosynthetic machinery and exhibited 4.23 and 1.72 times enhanced accumulation of Chl a and phycobiliproteins, respectively, which resulted in enhanced rate of photosynthesis (2.43 times) and respiration (1.38 times) against salt stress. Phycocyanin production in mutant cells was observed to enhance by 1.63 fold. Nitrogen metabolism plays a vital role in conferring halotolerance to cyanobacterial cells by influx of nitrate and efflux of Na+ ions from the cell. The NaCl tolerant mutant cells took up 2.29 times more nitrate as compared to the wild type and efficiently reduce it. Nitrate reductase and nitrite reductase activity in the mutant cells also improved by 2.45 and 2.31 times, respectively against salt stress. From these preliminary results, it could be deduced that enhanced nitrogen uptake and its efficient reduction might be a reason for adaptive and halotolerant behavior of the S. platensis mutant cells. Also, the NaCl tolerant mutant of S. platensis with significant improved growth and phycocyanin accumulation compared to the wild type can be commercially promising.

Keywords: chemical mutagenesis, NaCl tolerant mutant, nitrogen metabolism, photosynthetic machinery, phycocyanin

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Authors: Piya Roychoudhury, Ruma Pal

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Cyanobacteria, commonly known as blue green algae were found to be an effective bioreagent for nanoparticle synthesis. Nowadays silver nanoparticles (AgNPs) are very popular due to their antimicrobial and anti-proliferative activity. To exploit these characters in different biotechnological fields, it is very essential to synthesize more stable, non-toxic nano-silver. For this reason silver-gold alloy (Ag-AuNPs) nanoparticles are of great interest as they are more stable, harder and more effective than single metal nanoparticles. In the present communication we described a simple technique for rapid synthesis of biocompatible AgNP and Ag-AuNP employing cyanobacteria, Leptolyngbya and Lyngbya respectively. For synthesis of AgNP the biomass of Leptolyngbya valderiana (200 mg Fresh weight) was exposed to 9 mM AgNO3 solution (pH 4). For synthesis of Ag-AuNP Lyngbya majuscula (200 mg Fresh weight) was exposed to equimolar solution of hydrogen tetra-auro chlorate and silver nitrate (1mM, pH 4). After 72 hrs of exposure thallus of Leptolyngyba turned brown in color and filaments of Lyngbya turned pink in color that indicated synthesis of nanoparticles. The produced particles were extracted from the cyanobacterial biomass using nano-capping agent, sodium citrate. Firstly, extracted brown and pink suspensions were taken for Energy Dispersive X-ray (EDAX) analysis to confirm the presence of silver in brown suspension and presence of both gold and silver in pink suspension. Extracted nanoparticles showed a distinct single plasmon band (AgNP at 411 nm; Ag-Au NP at 481 nm) in Uv-vis spectroscopy. It was revealed from Transmission electron microscopy (TEM) that all the synthesized particles were spherical in nature with a size range of ~2-25 nm. In X-ray powder diffraction (XRD) analysis four intense peaks appeared at 38.2°, 44.5°, 64.8°and 77.8° which confirmed the crystallographic nature of synthesized particles. Presence of different functional groups viz. N-H, C=C, C–O, C=O on the surface of nanoparticles were recorded by Fourier transform infrared spectroscopy (FTIR). Scanning Electron microscopy (SEM) images showed the surface topography of metal treated filaments of cyanobacteria. The stability of the particles was observed by Zeta potential study. Antibiotic property of synthesized particles was tested by Agar well diffusion method against gram negative bacteria Pseudomonas aeruginosa. Overall, this green-technique requires low energy, less manufacturing cost and produces rapidly eco-friendly metal nanoparticles.

Keywords: cyanobacteria, silver nanoparticles, silver-gold alloy nanoparticles, spectroscopy

Procedia PDF Downloads 284

Authors: Benjamin Glasner, Carlos Henriquez, Fernando Alfaro, Nicole Trefault, Santiago Andrade, Rodrigo De La Iglesia

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A central question in ecology refers to the relative importance that local-scale variables have over community composition, when compared with regional-scale variables. In coastal environments, strong seasonal abiotic influence dominates these systems, weakening the impact of other parameters like micronutrients. After the industrial revolution, micronutrients like trace metals have increased in ocean as pollutants, with strong effects upon biotic entities and biological processes in coastal regions. Coastal picoplankton communities had been characterized as a cyanobacterial dominated fraction, but in recent years the eukaryotic component of this size fraction has gained relevance due to their high influence in carbon cycle, although, diversity patterns and responses to disturbances are poorly understood. South Pacific upwelling coastal environments represent an excellent model to study seasonal changes due to a strong influence in the availability of macro- and micronutrients between seasons. In addition, some well constrained coastal bays of this region have been subjected to strong disturbances due to trace metal inputs. In this study, we aim to compare the influence of seasonality and trace metals concentrations, on the community structure of planktonic picoeukaryotes. To describe seasonal patterns in the study area, satellite data in a 6 years time series and in-situ measurements with a traditional oceanographic approach such as CTDO equipment were performed. In addition, trace metal concentrations were analyzed trough ICP-MS analysis, for the same region. For biological data collection, field campaigns were performed in 2011-2012 and the picoplankton community was described by flow cytometry and taxonomical characterization with next-generation sequencing of ribosomal genes. The relation between the abiotic and biotic components was finally determined by multivariate statistical analysis. Our data show strong seasonal fluctuations in abiotic parameters such as photosynthetic active radiation and superficial sea temperature, with a clear differentiation of seasons. However, trace metal analysis allows identifying strong differentiation within the study area, dividing it into two zones based on trace metals concentration. Biological data indicate that there are no major changes in diversity but a significant fluctuation in evenness and community structure. These changes are related mainly with regional parameters, like temperature, but by analyzing the metal influence in picoplankton community structure, we identify a differential response of some plankton taxa to metal pollution. We propose that some picoeukaryotic plankton groups respond differentially to metal inputs, by changing their nutritional status and/or requirements under disturbances as a derived outcome of toxic effects and tolerance.

Keywords: Picoeukaryotes, plankton communities, trace metals, seasonal patterns

Procedia PDF Downloads 133

Authors: Hollie Ashworth, Sarah Heath, Nick Bryan, Liam Abrahamsen, Simon Kellet

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Sellafield has a number of fuel storage ponds, some of which have been open to the air for a number of decades. This has caused corrosion of the fuel resulting in a release of some activity into solution, reduced water clarity, and accumulation of sludge at the bottom of the pond consisting of brucite (Mg(OH)2) and other uranium corrosion products. Both of these phases are also present as colloidal material. 90Sr and 137Cs are known to constitute a small volume of the radionuclides present in the pond, but a large fraction of the activity, thus they are most at risk of challenging effluent discharge limits. Organic molecules are known to be present also, due to the ponds being open to the air, with occasional algal blooms restricting visibility further. The contents of the pond need to be retrieved and safely stored, but dealing with such a complex, undefined inventory poses a unique challenge. This work aims to determine and understand the sorption-desorption interactions of 90Sr and 137Cs to brucite and uranium phases, with and without the presence of organic molecules from chemical degradation and bio-organisms. The influence of organics on these interactions has not been widely studied. Partitioning of these radionuclides and organic molecules has been determined through LSC, ICP-AES/MS, and UV-vis spectrophotometry coupled with ultrafiltration in both binary and ternary systems. Further detailed analysis into the surface and bonding environment of these components is being investigated through XAS techniques and PHREEQC modelling. Experiments were conducted in CO2-free or N2 atmosphere across a high pH range in order to best simulate conditions in the pond. Humic acid used in brucite systems demonstrated strong competition against 90Sr for the brucite surface regardless of the order of addition of components. Variance of pH did have a small effect, however this range (10.5-11.5) is close to the pHpzc of brucite, causing the surface to buffer the solution pH towards that value over the course of the experiment. Sorption of 90Sr to UO2 obeyed Ho’s rate equation and demonstrated a slow second-order reaction with respect to the sharing of valence electrons from the strontium atom, with the initial rate clearly dependent on pH, with the equilibrium concentration calculated at close to 100% sorption. There was no influence of humic acid seen when introduced to these systems. Sorption of 137Cs to UO3 was significant, with more than 95% sorbed in just over 24 hours. Again, humic acid showed no influence when introduced into this system. Both brucite and uranium based systems will be studied with the incorporation of cyanobacterial cultures harvested at different stages of growth. Investigation of these systems provides insight into, and understanding of, the effect of organics on radionuclide partitioning to brucite and uranium phases at high pH. The majority of sorption-desorption work for radionuclides has been conducted at neutral to acidic pH values, and mostly without organics. These studies are particularly important for the characterisation of legacy wastes at Sellafield, with a view to their safe retrieval and storage.

Keywords: caesium, legacy wastes, organics, sorption-desorption, strontium, uranium

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Authors: N. K. Ambujam, V. Sudha

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Reservoirs are being contaminated all around the world with point source and Non-Point Source (NPS) pollution. The most common NPS pollutants are sediments and nutrients. Krishnagiri Reservoir (KR) has been chosen for the present case study, which is located in the tropical semi-arid climatic zone of Tamil Nadu, South India. It is the main source of surface water in Krishnagiri district to meet the freshwater demands. The reservoir has lost about 40% of its water holding capacity due to sedimentation over the period of 50 years. Hence, from the research and management perspective, there is a need for a sound knowledge on the spatial and seasonal variations of KR water quality. The present study encompasses the specific objectives as (i) to investigate the longitudinal heterogeneity and seasonal variations of physicochemical parameters, nutrients and biological characteristics of KR water and (ii) to examine the extent of degradation of water quality in KR. 15 sampling points were identified by uniform stratified method and a systematic monthly sampling strategy was selected due to high dynamic nature in its hydrological characteristics. The physicochemical parameters, major ions, nutrients and Chlorophyll a (Chl a) were analysed. Trophic status of KR was classified by using Carlson's Trophic State Index (TSI). All statistical analyses were performed by using Statistical Package for Social Sciences programme, version-16.0. Spatial maps were prepared for Chl a using Arc GIS. Observations in KR pointed out that electrical conductivity and major ions are highly variable factors as it receives inflow from the catchment with different land use activities. The study of major ions in KR exhibited different trends in their values and it could be concluded that as the monsoon progresses the major ions in the water decreases or water quality stabilizes. The inflow point of KR showed comparatively higher concentration of nutrients including nitrate, soluble reactive phosphorus (SRP), total phosphors (TP), total suspended phosphorus (TSP) and total dissolved phosphorus (TDP) during monsoon seasons. This evidently showed the input of significant amount of nutrients from the catchment side through agricultural runoff. High concentration of TDP and TSP at the lacustrine zone of the reservoir during summer season evidently revealed that there was a significant release of phosphorus from the bottom sediments. Carlson’s TSI of KR ranged between 81 and 92 during northeast monsoon and summer seasons. High and permanent Cyanobacterial bloom in KR could be mainly due to the internal loading of phosphorus from the bottom sediments. According to Carlson’s TSI classification Krishnagiri reservoir was ranked in the hyper-eutrophic category. This study provides necessary basic data on the spatio-temporal variations of water quality in KR and also proves the impact of point and NPS pollution from the catchment area. High TSI warrants a greater threat for the recovery of internal P loading and hyper-eutrophic condition of KR. Several expensive internal measures for the reduction of internal loading of P were introduced by many scientists. However, the outcome of the present research suggests for the innovative algae harvesting technique for the removal of sediment nutrients.

Keywords: NPS pollution, nutrients, hyper-eutrophication, krishnagiri reservoir

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Authors: Waqas Ali, Nadeem Majeed

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Requirement engineering is an essence of software development life cycle. The more time we spend on requirement engineering, higher the probability of success. Effective requirement engineering ensures and predicts successful software product. This paper presents the adaptation of requirement engineering practices in small and medium size companies of Pakistan. The study is conducted by questionnaires to show how much of requirement engineering models and practices are followed in Pakistan.

Keywords: requirement engineering, Pakistan, models, practices, organizations

Procedia PDF Downloads 683

Authors: Myunghoun Jang

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Most engineering colleges in South Korea have engineering design classes in order to develop and enhance a student's creativity and problem-solving ability. Many cases about engineering design class are shown in journals and magazines, but a case lasting many years is few. The engineering design class in the Department of Architectural Engineering, Jeju National University was open in 2009 and continues to this year. 3-5 teams in every year set up their problems found their solutions and produced good results. Three of the results obtained patents. The class also provides students with opportunities to improve communication skill because they have many discussions in solving their problems.

Keywords: engineering design, architectural engineering, team-based learning, construction safety

Procedia PDF Downloads 199