Insights into the growth and biochemical defense responses associated with fenitrothion toxicity and uptake by freshwater cyanobacteria.

The extensive use of fenitrothion (FNT) in agricultural practices induces its persistence in soil and waterways. Therefore, it is essential to implement effective management practices such as using cyanobacteria for FNT removal and accumulation, particularly under accidental contamination. To this end, we evaluated the responses of two freshwater cyanobacteria taxa, Nostoc muscorum and Anabaena laxa to mild (7.5 mg L-1) and high (15 mg L-1) levels of FNT over a period of 7 d. Compared to N. muscorum, A. laxa was more tolerant to FNT, exhibiting higher FNT uptake and removal efficiencies at mild (16.3%) and high (17.5%) levels. FNT induced a dose-dependent decrease in cell growth, Chl a, phosphoenolpyruvate carboxylase and ribulose-1,5-bisphosphate carboxylase/oxygenase activities, which were more pronounced in N. muscorum. Moreover, FNT significantly increased oxidative damage markers i.e., increased lipid peroxidation (MDA), protein oxidation, H2O2 levels and NADPH oxidase enzyme activity, to more extent in N. muscorum. Compared to N. muscorum, A. laxa had high antioxidant capacity (FRAP), glutathione and increased activities of glutathione-S-transferase, glutathione reductase, glutathione peroxidase and superoxide dismutase, suggesting a robust antioxidant defense mechanism to mitigate FNT toxicity. However, N. muscorum devoted the induction of ascorbate content and the activity of catalase, peroxidase, monodehydroascorbate reductase, ascorbate peroxidase, and dehydroascorbate reductase enzymes. Although A. laxa had greater intracellular FNT, it experienced less FNT-induced oxidative stress, likely due to over production of antioxidants. Consequently, A. laxa is considered as a promising candidate for FNT phycoremediation. Our findings provide fundamental information on species-specific toxicity of FNT among cyanobacteria and the environmental risk of FNT toxicity in aquatic environments.

Synergistic regulation of hydrogen sulfide and nitric oxide on biochemical components, exopolysaccharides, and nitrogen metabolism in nickel stressed rice field cyanobacteria.

The present study examined the regulatory mechanism of hydrogen sulfide (H2S) and nitric oxide (NO) in nickel (Ni) stressed cyanobacteria viz., Nostoc muscorum and Anabaena sp. by analyzing growth, photosynthetic pigments, biochemical components (protein and carbohydrate), exopolysaccharides (EPS), inorganic nitrogen content, and activity of enzymes comprised in nitrogen metabolism and Ni accumulation. The 1 µM Ni substantially diminished growth by 18% and 22% in N. muscorum and Anabaena sp. respectively, along with declining the pigment contents (Chl a/Car ratio and phycobiliproteins), and biochemical components. It also exerted negative impacts on inorganic uptake of nitrate and nitrite contents; nitrate reductase and nitrite reductase; and ammonium assimilating enzymes (glutamine synthetase, glutamate synthase, and glutamate dehydrogenase exhibited a reverse trend) activities. Nonetheless, the adverse impact of Ni can be mitigated through the exogenous supplementation of NaHS [sodium hydrosulfide (8 µM); H2S donor] and SNP [sodium nitroprusside (10 µM); NO donor] which showed substantial improvement on growth, pigments, nitrogen metabolism, and EPS layer and noticeably occurred as a consequence of a substantial reduction in Ni accumulation content which minimized the toxicity effects. The accumulation of Ni on both the cyanobacterial cell surface (EPS layer) are confirmed by the SEM-EDX analysis. Further, the addition of NO scavenger (PTIO; 20 µM) and inhibitor of NO (L-NAME; 100 µM); and H2S scavenger (HT; 20 µM) and H2S inhibitor (PAG; 50 µM) reversed the positive responses of H2S and NO and damages were more prominent under Ni stress thereby, suggesting the downstream signaling of H2S on NO-mediated alleviation. Thus, this study concludes the crosstalk mechanism of H2S and NO in the mitigation of Ni-induced toxicity in rice field cyanobacteria.

Biofabrication of Silver Nanoparticles Using Nostoc muscorum Lukesova 2/91: Optimization, Characterization, and Biological Applications.

Purpose: The biological synthesis of nanoparticles (NPs) has become a new methodology for the eco-friendly production of NPs with high scalability and biocompatibility. Cyanobacteria are one of the most widespread microorganisms on Earth and have been proven to be successful biofactories for synthesizing NPs. It is challenging to discover new microalgae with the potential to synthesize NPs of small size with high stability. Methods: Nostoc muscorum Lukesova 2/91 was isolated, purified, and identified morphologically and genetically using microscopy and DNA sequencing. Volatile biomolecules in aqueous algal extracts were assessed using gas chromatography-mass spectroscopy (GC-MS). Results: Data showed that the main biomolecules were fatty acids and their esters, followed by secondary metabolites. Algal extract was used to convert silver nitrate (AgNO3) into silver NPs under various optimized parameters. 1 mM of AgNO3, 1:1 (V/V ratio of algal extract to AgNO3), 25 °C, under light illumination, for 24 h, at pH 7.4 were the optimum conditions for NP production (Nos@AgNPs). Nos@AgNPs were characterized using UV-VIS spectroscopy, FTIR, TEM, SEM, EDx, mapping, and a Zetasizer. The wavelength of Nos@AgNPs was 401.4 nm and their shapes were cubic to oval, with an average diameter of 11.8 ± 0.5 nm. FTIR spectroscopy revealed that proteins/polysaccharides could be the main reductants, whereas these molecules and/or fatty acids could be stabilizers for NP synthesis. Nos@AgNPs (86.15%) was silver and had a hydrodynamic diameter of 10.7 nm with a potential charge of -19.7 mV. Antiproliferative and antimicrobial activities of Nos@AgNPs were evaluated. Nos@AgNPs exhibited significant inhibitory activity against lung, colon, and breast cancer cells and considerable biocidal activity against Staphylococcus aureus, Escherichia coli, Klebsiella pneumonia, and Pseudomonas aeruginosa. Conclusion: N. muscorum Lukesova 2/91 is an excellent source for the biofabrication of small and stable AgNPs with potent inhibitory effects against cancer and bacterial cells.

Cyanobacterial degradation of herbicide 2,4-dichlorophenoxyacetic acid (2,4-D): Its response to the oxidative stress induced by the primary degradation product 2,4-dichlorophenol (2,4-DCP).

Excessive use of herbicides in agricultural fields has become a major environmental concern due to the negative effects on the ecosystem. Microbial degradation has been well-known as an effective approach for combating such non-natural substances in soil. In the present study, the degradation of 2,4-Dichlorophenoxyacetic acid (2,4-D) as a result of metabolic activities of a cyanobacterium Nostoc muscorum Meg 1 was investigated using GC-MS analysis. After seven days of 2,4-D exposure, the main residue obtained was 2,4-dichlorophenol (2,4-DCP) at RT: 8.334 (confirmed using NIST library). The effects of 2,4-DCP were studied in a cyanobacterium Nostoc muscorum Meg 1 isolated from a rice field where 2,4-D is commonly used. Exposure to 2,4-DCP at 20, 40, and 80 ppm significantly increased ROS production in the cyanobacterium by 74, 107, and 211 % (p < 0.001). With rising 2,4-DCP concentrations in the surroundings, lipid peroxidation and protein oxidation in the organism correspondingly increased, indicating cellular injury. The mRNA and protein contents, and also the activities of different oxidant neutralizing enzymes such as CAT, SOD, GR, and GPx and the non-enzymatic antioxidants (proline, GSH, thiol and phytochelatin content) were found augmented in 20 ppm 2,4-DCP exposed cultures. However, in the presence of 40 and 80 ppm 2,4-DCP, most enzymatic and non-enzymatic antioxidants were severely compromised. At higher exposures, the organism's attempt to mitigate the oxidants was still visible, as both proline and TSH levels increased. SEM and TEM analysis aided in visualizing the effects of 2,4-DCP on the morphology and ultrastructures of the organism.

Nostoc muscorum is a novel source of microalgal lectins with potent antiviral activity against herpes simplex type-1.

In our survey for a new antiviral agent, two types of lectin were purified from Nostoc muscorum using both ion-exchange and affinity columns chromatography. Nostoc muscorum lectins (NMLs) are categorized based on their carbohydrate preference. Nostoc muscorum lectin-1(NML-1) exhibited a strict binding specificity for complex glycoproteins without linked carbohydrates, and the other displayed specificity for α- glycosides mannose polymers (NML-2) and was classified as a glycoprotein with 16.8% linked carbohydrates. NML-1 displayed a single band of 166 kDa on native-PAGE and two bands of 81 kDa and 85 kDa on SDS-PAGE, which confirmed the heterodimeric nature of this lectin. While NML-2 is a 50 kDa glycoprotein composed of 25 kDa subunits. Physical characterization of NML-1 displayed its stability at a higher temperature of 90 °C for 5 min and over a wide pH range (4-9), while MNL-2 displayed stability up to a temperature of 80 °C for 25 min and a pH range of 5-8. NML-1 didn't require metal ions for agglutination activity, while the activity of NML-2 was doubled by manganese ions. The antiviral activity of two lectins was assessed against herpes simplex type-1 (HSV-1) using a plaque assay which revealed that NML-1 inhibited HSV-1 infection at an early stage in contrast to NML-2 which exerted its antiviral effect at the late stage of infection. These results suggest that Nostoc muscorum is a unique lead for antiviral drug discovery as it is a novel source for antiviral lectins with different modes of action.

A biochemical, physiological and molecular evaluation of how the herbicide 2, 4-dichlorophenoxyacetic acid intercedes photosynthesis and diazotrophy in the cyanobacterium Nostoc muscorum Meg 1.

Among the non-target microorganisms residing in crop fields that are potentially vulnerable to herbicides are cyanobacteria. They contribute to the maintenance of soil quality and fertility and hence are considered to be an important component of soil microflora. Consequently, the present study was aimed to check the influence of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) on some major parameters of carbon (CO2) and nitrogen (N2) fixations of a cyanobacterium Nostoc muscorum Meg 1 isolated from a rice field in Cherrapunji, Meghalaya, India. These include various photosynthetic pigments, the oxygen-evolving complex activity of the PSII, the protein contents of RuBisCO, D1 protein, isocitrate dehydrogenase (IDH), nitrogenase and glutamine synthetase (GS) enzymes, the heterocyst percentage, nitrogenase and GS enzyme activities, and production of total proteins and carbohydrates in the cyanobacterium in a varying range of 50 to 125 ppm doses of 2,4-D. The mRNA levels of several proteins were also analyzed. Besides carotenoid concentration that enhanced at 50 ppm, all other parameters were compromised by 2,4-D in a dose-dependent manner resulting in a reduction in photosynthetic and N2-fixing activities. The negative effect on N2-fixation was partly due to compromised IDH activity. RT-PCR analysis further showed that these negative effects were initiated at transcription levels as mRNA contents of all enzymes studied were found compromised under 2,4-D exposure. The scanning and transmission electron microscopy further revealed herbicide induced adverse changes in the morphology and ultrastructure of the organism. The significance of the work lies in its detailed analysis of the effect of 2,4-D at biochemical, physiological, and molecular levels.

Arsenate and arsenite-induced inhibition and recovery in two diazotrophic cyanobacteria Nostoc muscorum and Anabaena sp.: study on time-dependent toxicity regulation.

Exposure time, metal bio-accumulation, and upregulation of ascorbate-glutathione (AsA-GSH) cycle are the key factor that provide tolerance against heavy metal stress. Thus, the current study is an endeavor to prove our hypothesis that regulation of arsenate (AsV: 50, 100, and 150 mM) and arsenite (AsIII: 50, 100, and 150 µM) toxicity is time dependent (48-96 h) due to modulation in bio-accumulation pattern, AsA-GSH cycle, and non-enzymatic antioxidants in two paddy field cyanobacteria Nostoc muscorum ATCC27893 and Anabaena sp. PCC7120. After 48 h, reduction in growth associated with increased sensitivity index, As bio-accumulation, and oxidative stress was observed which further intensified after 96 h but the degree of damage was lesser than 48 h. It denotes a significant recovery in growth after 96 h which is correlated with decreased As bio-accumulation and oxidative stress due to increased efficiency of AsA-GSH cycle and non-enzymatic antioxidants. Both the species of As caused significant rise in oxidative biomarkers as evident by in -vitro analysis of O2·-, H2O2, and MDA equivalent contents despite appreciable rise in the activity antioxidative enzymes APX, DHAR, and GR. The study concludes that among both forms of arsenic, AsIII induced more toxic effect on growth by over-accumulating the ROS as evident by weak induction of AsA-GSH cycle to overcome the stress as compared to AsV. Further, with increasing the time exposure, apparent recovery was noticed with the lower doses of AsV, i.e., 50 and 100 mM and AsIII, i.e., 50 and 100 µM; however, the toxicity further aggravated with higher dose of both AsV and AsIII. Study proposes the deleterious impact of AsV and AsIII on cyanobacteria N. muscorum and Anabaena sp. but the toxicity was overcome by time-dependent recovery.

Modulation in isocitrate dehydrogenase activity under citrate enrichment affects carbon and nitrogen fixations in the cyanobacterium Nostoc muscorum Meg 1.

The enzyme isocitrate dehydrogenase (IDH) converts isocitrate synthesized from citrate to α-ketoglutarate in the TCA cycle. In cyanobacteria, α-KG has an additional role where it donates its carbon skeleton for ammonium assimilation in the GS-GOGAT pathway thereby linking carbon and nitrogen metabolisms. Looking at this crucial function of IDH that makes α-KG available for both carbon and nitrogen assimilation, changes brought about in its activity under excess availability of citrate in a cyanobacterium was evaluated. Further, how these changes are transmitted downstream affecting carbon and nitrogen metabolisms were also evaluated. A 100 µM citrate supplementation induced IDH activity. Consequently, there was an increase in concentrations of photosynthetic pigments, D1 protein and RuBisCO as well as in PSII activity. Heterocyst differentiation was initiated and an upsurge in the activities of nitrogenase and GS were recorded. An enhancement in the total protein and carbohydrate content reiterated the positive influence of citrate enrichment on carbon and nitrogen fixation. The increase in the mRNA contents of IDH, D1 protein, RuBisCO, nitrogenase and GS indicated their induction at the genetic level. Finally, there was augmentation in total biomass production by ∼28%. Interestingly as citrate concentration was increased to 500 µM, both C- and N- fixations were highly compromised suggesting that even though citrate is an essential metabolite in the cells, it became toxic beyond a certain concentration to the organism. SEM and TEM studies showed no changes in the organism's morphology and ultra-structure in presence of 100 µM citrate while adverse changes were noticed in presence of 500 µM citrate.

Regulation of redox homeostasis in cadmium stressed rice field cyanobacteria by exogenous hydrogen peroxide and nitric oxide.

In the present study, defensive strategies of H2O2 mediated NO signaling were analyzed in Cd stressed Nostoc muscorum and Anabaena sp. Exogenously supplied SNP (10 µM) and H2O2 (1 µM) lessen the toxicity of Cd (6 µM) but without NO; H2O2 was unable to release the stress from cyanobacterial cells potentially. The reduced contents of exopolysaccharide, protein content, endogenous NO and enzymatic antioxidants (SOD, POD, CAT, and GST) due to Cd toxicity, were found increased significantly after exogenous application of H2O2 and SNP thereafter, cyanobacterial calls flourished much better after releasing toxic level of Cd. Moreover, increased level of ROS due to Cd stress also normalized under exogenous application of H2O2 and SNP. However, chelation of NO hindered the signaling mechanism of H2O2 that diminished its potential against Cd stress while signaling of NO has not been hindered by chelation of H2O2 and NO potentially released the Cd stress from cyanobacterial cells. In conclusion, current findings demonstrated the synergistic signaling between H2O2 and NO towards the improvement of cyanobacterial tolerance to Cd stress, thereby enhancing the growth and antioxidant defense system of test cyanobacteria that improved fertility and productivity of soil even under the situation of metal contamination.

Biogas-based production of glycogen by Nostoc muscorum: Assessing the potential of transforming CO2 into value added products.

The potential of the filamentous N2-fixing cyanobacterium Nostoc muscorum for CO2 capture from high-loaded streams (i.e. flue gas or biogas) combined with the accumulation of glycogen (GL) and polyhydroxybutyrate (PHB), was evaluated under nutrient-sufficient and nutrient-limited conditions. N. muscorum was able to grow under CO2 contents from 0.03 up to 30% v/v, thus tolerating CO2 concentrations similar to those found in raw biogas or flue-gas, with maximum CO2-fixation rates of 191.9 ± 46 g m-3 d-1 at a biomass concentration of 733.3 ± 207.4 mg TSS L-1. Despite N. muscorum was inhibited by the presence of H2S, the co-inoculation with activated sludge resulted in both CO2 and H2S depletion. Moreover, N. muscorum accumulated GL up to ∼54% dcw under N and P-deprivation, almost 36 times higher than that recorded under nutrients sufficient condition. The addition of 10% extra carbon in the form of valeric acid not only did not hamper the growth of N. muscorum (336.0 ± 113.1 mg TSS L-1) but also increased the GL content to ∼58% dcw. On the contrary, a negligible PHB accumulation was found under the tested conditions, likely due to the high CO2 concentration of 30% v/v in the headspace and therefore the high availability of inorganic carbon for the cultures. N. muscorum cultures achieved VFAs degradations up to ∼78% under controlled pH. These results supported N. muscorum as a sustainable alternative for CO2-capture and greenhouse gas mitigation or for photosynthetic biogas upgrading coupled with value added biomass production.

Analyzing dose dependency of antioxidant defense system in the cyanobacterium Nostoc muscorum Meg 1 chronically exposed to Cd2.

The aim of the present study was to analyze the dose dependency of oxidant-antioxidant homeostasis in Cd2+ exposed Nostoc muscorum Meg 1 cells. Quantification of percent DNA loss, protein oxidation and lipid peroxidation was carried out to assess Cd2+ induced ROS mediated damages to the organism. The countermeasures adopted by the cyanobacterium were also evaluated by computing various components of both enzymatic and non-enzymatic antioxidants. Exposure to different Cd2+ (0.1, 0.2, 0.3, 0.5, 1, 1.5, 2, 2.5, 3 ppm) doses showed substantial increase in ROS content in the ranges of 20-181% and 116-323% at the end of first and seventh day. The DNA damage, protein oxidation and lipid peroxidation were increased by 11-62%, 7-143% and 13-183% with increasing Cd2+ concentrations at the end of seven days. TEM images clearly showed damages to the cell wall, cell membrane and thylakoid organization at higher Cd2+ (0.5-3 ppm) concentrations. Cd2+ exposure up to 0.5 ppm registered increase in contents of antioxidative enzymes (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR)) and in non-enzymatic antioxidants (glutathione, total thiol, phytochelatin and proline) indicating stimulation of ROS mitigating machinery. However, toxicity of Cd2+ was evident as at higher concentrations the cellular morphology and ultra-structures were negatively affected and the capacities of the cells to generate various antioxidant measures were highly compromised. The organism registered 96-98% sorption ability from a solution supplemented with 0.3 ppm Cd2+ and thus show realistic potential as Cd2+ bioremediator in wastewater treatment.

Cytokinin alleviates cypermethrin toxicity in Nostoc muscorum by involving nitric oxide: Regulation of exopolysaccharides secretion, PS II photochemistry and reactive oxygen species homeostasis.

Growth of the most important nitrogen fixing cyanobacterium Nostoc muscorum is reported to be badly affected by the application of insecticides. To overcome their damaging effects, several strategies are being used. Out of these, some works on kinetin (KN, a synthetic cytokinin) has been recognized that it can overcome toxicity of insecticides in cyanobacteria. Besides this, it is now known that every hormone needs certain second messengers such as nitric oxide (NO) for its action. But implication of NO in KN-mediated regulation of insecticide toxicity is yet to be investigated. Hence in the current study, we have investigated the possible involvement of NO in KN-mediated regulation of cypermethrin toxicity in the cyanobacterium Nostoc muscorum. Cypermethrin decreased growth of Nostoc muscorum which was accompanied by decreased pigment contents and altered photosystem II (PS II) photochemistry that resulted in inhibition of photosynthetic process but KN significantly ameliorated cypermethrin toxicity. Cypermethrin induced production of free radicals (in-vivo and in-vitro) and weakened defensive mechanism (enzymatic and non-enzymatic defense system) which was restored by KN. Further, the results revealed that NG-nitro-l-arginine methyl ester (l-NAME, an inhibitor of nitric oxide synthase) worsened the effect of cypermethrin toxicity even in the presence of KN while 2-4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO, a scavenger of NO) reversed KN-mediated amelioration even in the presence of sodium nitroprusside (SNP, an NO donor), suggesting that endogenous NO is required for mitigation of cypermethrin toxicity. Overall, our results first time show that endogenous NO is essential for KN-mediated mitigation of cypermethrin toxicity in the Nostoc muscorum.

Toxicity of NSAID drug (paracetamol) to nontarget organism-Nostoc muscorum.

Due to many folds increase in application of human and veterinary medicines, pharmaceuticals, a new category of pollutants, have emerged in our environment. They exist as residues in rivers, sewage effluents, streams, surface, ground, and potable water. Paracetamol (acetaminophen) is one such drug that is used as an antipyretic and analgesic medicine. It is a non-steroidal antiinflammatory drug (NSAID) and is easily available in the market because no medical prescription is necessary for its purchase and use. Paracetamol remains physiologically active even after their expiry period. Their detection in the environment in bioactive form has resulted in adverse effects on nontarget species. To determine the effect of paracetamol on aquatic photosynthetic organic (Cyanobacteria-Nostoc muscorum), present study was performed. Paracetamol (25 mg/L, 50 mg/L, 75 mg/L, 100, 125, and 150 mg/L) exposure showed toxic responses on the test organism by generating oxidative stress (MDA, H2O2, O2.-). Paracetamol caused a significant decrease in growth of cyanobacteria and showed EC50 113.68 mg/L after the 6th day of treatment. Photosynthetic pigments (chlorophyll, carotenoid, and phycobiliprotein) decreased with paracetamol increase. Antioxidant enzymatic (SOD, CAT, APX, GST, and GR) and osmolyte (Proline) also increased with increase in paracetamol to counteract the oxidative stress.

Unstandardized UV-C dose used for killing harmful cyanobacteria may instead initiate accelerated growth in the target organisms.

Although UV-C radiation has been in use for killing unwanted cyanobacteria, experiments with lower doses of UV-C radiation instead showed induction of growth related parameters and enhanced biomass production in the cyanobacterium Nostoc muscorum Meg1. When the cyanobacterial cultures were exposed to UV-C radiation of varying doses (6, 12 and 18 mJ/cm2), concentrations of various photo-absorbing pigments, RuBisCO and D1 protein of PSII; activities of oxygen evolving complex, nitrogenase and glutamine synthetase were significantly increased upon 6 and 12 mJ/cm2 UV-C radiation exposures. Resulting higher photosynthetic performance was evident from the augmentation in carbohydrate content by ∼49% under single exposure to 6 mJ/cm2 UV-C by fifteenth day. The increased performances of both RuBisCO and D1 proteins were in part also due to induction at the genetic level as seen from the increase in their mRNA and protein levels under treatment. Similar increase was also observed in protein (16%) and in lipid contents (43%) that reflected an upsurge in the total biomass. Highest biomass (463 mg/L/d) was noted in culture exposed to 6 mJ/cm2 UV-C radiation, representing a ∼25% increase. Furthermore the possibility of this organism using part of the incident UV-C radiation as an additional source of energy was deduced from an experiment where the thylakoid membranes excited within UV (226-400 nm) range showed emission at longer wavelengths with an emission maximum at ∼640 nm. Thus this work provides evidence that lower UV-C doses can potentially augment cyanobacterial growth and use of unstandardized UV-C doses for restricting cyanobacterial growth may in fact produce contrary result.

Modulation of oxidant and antioxidant homeostasis in the cyanobacterium Nostoc muscorum Meg1 under UV-C radiation stress.

The present work was conducted to study how restoration of perturbed oxidant and antioxidant homeostasis is achieved in the UV-C radiation exposed cells of cyanobacterium Nostoc muscorum Meg1. Exposure to varying doses of UV-C radiation (6, 12, 18 and 24 mJ/cm2) showed damage to ultrastructures especially cytoplasmic membrane, cell wall and organisation of thylakoid membranes of the cyanobacterium under transmission electron microscope (TEM). All doses of UV-C exposure significantly induced most of the enzymatic antioxidant {catalase, superoxide dismutase (SOD) and glutathione reductase (GR)} activities, their protein levels (western blot analysis) and mRNA levels (real time PCR analysis) within the first hour of post UV-C radiation incubation period. In the same way, contents of many non-enzymatic antioxidants such as ascorbic acid, reduced glutathione, proline, phenol and flavonoids were also augmented in response to such UV-C radiation exposure. Although notable increase in ROS level was only seen in cultures treated with 24 mJ/cm2 UV-C exposure which also registered increase in protein oxidation (22%) and lipid peroxidation (20%), this boost in both enzymatic and non-enzymatic antioxidants was significant in all radiation exposed cells indicating cell's preparation to combat rise in oxidants. Further, albeit all antioxidants increased considerably, their levels were restored back to control values by day seventh re-establishing physiological redox state for normal metabolic function. The combined efficiency of the enzymatic and non-enzymatic antioxidants were so effective that they were able to bring down the increase levels of ROS, lipid peroxidation and protein oxidation to the physiological levels within 1 h of radiation exposure signifying their importance in the defensive roles in protecting the organism from oxidative toxicity induced by UV-C radiation exposure.

Physiological and biochemical responses to aluminum-induced oxidative stress in two cyanobacterial species.

Phycoremediation technologies significantly contribute to solving serious problems induced by heavy metals accumulation in the aquatic systems. Here we studied the mechanisms underlying Al stress tolerance in two diazotrophic cyanobacterial species, to identify suitable species for Al phycoremediation. Al uptake as well as the physiological and biochemical responses of Anabaena laxa and Nostoc muscorum to 7 days Al exposure at two different concentrations i.e., mild (100 µM) and high dose (200 µM), were investigated. Our results revealed that A. laxa accumulated more Al, and it could acclimatize to long-term exposure of Al stress. Al induced a dose-dependent decrease in photosynthesis and its related parameters e.g., chlorophyll content (Chl a), phosphoenolpyruvate carboxylase (PEPC) and Ribulose‒1,5‒bisphosphate carboxylase/oxygenase (RuBisCo) activities. The affect was less pronounced in A. laxa than N. muscorum. Moreover, Al stress significantly increased cellular membrane damage as indicated by induced H2O2, lipid peroxidation, protein oxidation, and NADPH oxidase activity. However, these increases were lower in A. laxa compared to N. muscorum. To mitigate the impact of Al stress, A. laxa induced its antioxidant defense system by increasing polyphenols, flavonoids, tocopherols and glutathione levels as well as peroxidase (POX), catalase (CAT), glutathione reductase (GR) and glutathione peroxidase (GPX) enzymes activities. On the other hand, the antioxidant increases in N. muscorum were only limited to ascorbate (ASC) cycle. Overall, high biosorption/uptake capacity and efficient antioxidant defense system of A. laxa recommend its feasibility in the treatment of Al contaminated waters/soils.

Comparative studies on growth and Pb(II) removal from aqueous solution by Nostoc muscorum and Anabaena variabilis.

Industrial wastewater containing heavy metals is a major environmental problem that needs to be treated. This study reported the ability of two fresh water algae cyanobacteria (Nostoc muscorum and Anabaena variabilis) to remove lead from aqueous solutions of four different initial concentrations (0-50 mg/L-1) for 21 days under controlled laboratory conditions. Results obtained in this study showed a maximum removal of Pb(II) (97.8%) by N. muscorum at 15 mg/L-1 initial metal concentration however the maximum removal by A. variabilis at the same concentration was 71.4% after 16 day of incubation. These N. muscorum appeared to be more efficient than A. variabilis for removing Pb(II). Algal growth, pigments in the algae cells were measured during incubation period. Lower concentrations of lead increased biomass, OD, chlorophyll a and carotenoids in both algae. On the other hand, higher concentrations of lead were inhibitory for growth.

Microalgal cultivation for biofertilization in rice plants using a vertical semi-closed airlift photobioreactor.

Nitrogen (N) is one of the most important limiting factors in conventional rice (Oryza sativa) production, which heavily relies on synthetic fertilizers. In this study, we researched on the development and use of a vertical semi-closed airlift photobioreactor (PBR) for microalgal cultivation and subsequently determined the efficacy of microalgae-based fertilizers to rice plant growth. The PBR system was developed to produce two strains of N2-fixing cyanobacteria (Anabaena sp. UTEX 2576, Nostoc muscorum UTEX 2209S), and a polyculture of Chlorella vulgaris (UTEX 2714) and Scenedesmus dimorphus (UTEX 1237). When these biofertilizers were evaluated for rice under the greenhouse conditions, results showed that the rice plant heights treated with polyculture-based microalgal biomass were similar to or better than the urea treatment. The effects of the inoculation of the N2-fixing cyanobacterial inoculation on seedling growth was not statistically significant. In conclusion, the vertical semi-closed system PBR cultivation method developed in this study proved to be a simple and effective method for cultivating microalgae. Demonstration of the reliable production system for N2-fixing cyanobacteria and chlorophytes at a medium scale could potentially open the future application of microalgal biofertilizers in rice production.

Kinetin alleviates chromium toxicity on growth and PS II photochemistry in Nostoc muscorum by regulating antioxidant system.

The present study was undertaken to evaluate the metal toxicity alleviating effects of kinetin (KN, 10 nM) on growth, photosynthetic pigments and photochemistry of PS II in the cyanobacterium Nostoc muscorum exposed to chromium (CrVI) stress (100 and 150 µM). Chromium declined growth, photosynthetic pigments (chlorophyll a, phycocyanin and carotenoids), photosynthetic oxygen evolution rate and parameters of fluorescence kinetics (ϕP0, FV/F0, ϕE0, Ψ0 and PIABS except F0/FV) in concentration dependent manner, while stimulating effects on respiration, energy flux parameters (ABS/RC, TR0/RC, ET0/RC and DI0/RC), oxidative stress biomarkers i.e., superoxide radical (SOR), hydrogen peroxide (H2O2) and lipid peroxidation (TBARS contents) and antioxidative enzymes: superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and glutathione-S-transferase (GST), were observed. However, upon addition of KN in the growth medium an alleviating effect against chromium induced toxicity on growth, photosynthetic pigments and photochemistry of PS II was recorded. This had occurred due to substantial reduction in levels of oxidative stress biomarkers: SOR, H2O2 and TBARS contents with concomitant rise in activity of antioxidative enzymes: SOD, POD, CAT and GST and appreciable lowering in the cellular accumulation of chromium. The overall results demonstrate that KN application significantly alleviated chromium induced toxicity on growth performance of the cyanobacterium N. muscorum due to significant improvement in photosynthetic pigments and photochemistry of PS II by up-regulating the activity of antioxidative enzymes, and declining cellular accumulation of chromium. Furthermore, Cr induced toxicity at lower dose (100 µM) was found to be ameliorated more efficiently in N. muscorum following supplementation of KN.

Toxicity assessment of arsenate and arsenite on growth, chlorophyll a fluorescence and antioxidant machinery in Nostoc muscorum.

The present study deals with impact of varied doses of arsenite (AsIII; 50, 100 and 150 µM) and arsenate (AsV; 50, 100 and 150 mM) on growth, photosynthetic pigments, photochemistry of photosystem II, oxidative biomarkers, (O2•¯, H2O2 and MDA equivalents contents) and activity of antioxidant enzymes in diazotrophic cyanobacterium Nostoc muscorum after 48 and 96 h of the treatments. The reduction in growth, pigment contents (Chl a, Phy and Car) and PS II photochemistry was found to increase with enhanced accumulation of test metal in cells, and the damaging effect on photosynthetic pigments showed the order (Phy > chl a> Car). The negative effect on PS II photochemistry was due to significant decrease in the value of JIP kinetics ϕP0, FV/F0, ϕE0,Ψ0 and PIABS except F0/FV and significant rise in values of energy flux parameters such as ABS/RC, TR0/RC, ET0/RC and DI0/RC. Both the species of arsenic caused significant rise in oxidative biomarkers as evident by in vitro and in vivo analysis of (O2•¯, H2O2 and MDA equivalents contents) despite of appreciable rise in the activity antioxidative enzymes such as SOD, POD, CAT and GST. The study concludes that in among both forms of arsenic, arsenite effect was more dominant on growth, photosynthetic pigments; oxidative stress biomarkers as evident by weak induction of anti-oxidative defense system to overcome the stress as compared to arsenate.