Shruti Pavagadhi

Removal of microcystin-LR and microcystin-RR by graphene oxide: Adsorption and kinetic experiments

Graphene oxide (GO) was employed in the present study for removal of two commonly occurring algal toxins, microcystin-LR (MC-LR) and microcystin-RR (MC-RR), from water. The adsorption performance of GO was compared to that of commercially available activated carbon. Further, adsorption experiments were conducted in the presence of other environmental pollutants to understand the matrix effects of contaminated water on the selective adsorption of MC-LR and MC-RR onto GO. The environmental pollutants addressed in this study included different anions (nitrate NO3-, nitrite NO2-, sulphate SO4(2-), chloride (Cl(-)), phosphate PO4(3-) and fluoride (F(-))) and cations (sodium (Na(+)), potassium (K(+)), magnesium (Mg(2+)) and calcium (Ca(2+))). GO showed very a high adsorption capacity of 1700 μg/g for removal of MC-LR and 1878 μg/g for MC-RR while the maximum adsorption capacity obtained with the commercial activated carbon was 1481.7 μg/g and 1034.1 μg/g for MC-LR and MC-RR, respectively. The sorption kinetic experiments revealed that more than 90% removal of both MC-LR/RR was achieved within 5 min for all the doses studied (500, 700 and 900 μg/L). GO could be reused as an adsorbent following ten cycles of adsorption/desorption with no significant loss in its adsorption capacity.

Toxicological evaluation of microcystins in aquatic fish species: current knowledge and future directions.

Microcystins (MCs) are algal toxins produced intracellularly within the algal cells, and are subsequently released into the aquatic systems. An increase in the frequency and intensity of occurrence of harmful algal blooms has directed the global attention towards the presence of MCs in aquatic systems. The effects of MCs on fish have been verified in a number of studies including histological, biochemical and behavioral effects. The toxicological effects of MCs on different organs of fish are related to the exposure route (intraperitoneal injection, feeding or immersion), the mode of uptake (passive or active transport) as well as biotransformation and bioaccumulation capabilities by different organs. This paper reviews the rapidly expanding literature on the toxicological evaluation of MCs in fish from both field studies and controlled laboratory experimental investigations, integrates the current knowledge available about the mechanisms involved in MC-induced effects on fish, and points out future research directions from a cross-disciplinary perspective. In addition, the need to carry out systematic fish toxicity studies to account for possible interactions between MCs and other environmental pollutants in aquatic systems is discussed.

Experimental studies on removal of microcystin-LR by peat

Cyanotoxins have caused worldwide concerns for their eclectic occurrence and toxic effects, which led to an intensive search of cost-effective techniques for their removal from contaminated waters. A range of biomaterials was tested for their efficacy to adsorb a potent cyanotoxin, microcystin-LR (MCLR). Among these sorbents, peat showed the maximum efficacy to sequester MCLR. The BET (Brunauer-Emmett-Teller) surface area of peat was found to be 12.134 m(2)/g. The pH of the reaction media played a significant role in removal of MCLR; maximum adsorption occurred at pH 3. Kinetic studies showed that the adsorption of MCLR onto peat was a rapid process. The adsorption capacity (Q(max)) from the Langmuir model was found to be 255.7 μg/g at pH 3. Among various desorption media studied, strong alkali (2N NaOH) showed highest desorption (94%).

Biochemical response of diverse organs in adult Danio rerio (zebrafish) exposed to sub-lethal concentrations of microcystin-LR and microcystin-RR: a balneation study.

The present study was carried out to examine the dose-response of microcystin-LR (MC-LR) and microcystin-RR (MC-RR) toxicity in adult Danio rerio (zebrafish) under balneation conditions at various time points. The differential responses of superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione-S-transferase (GST) as biomarkers were assessed for oxygen mediated toxicity in liver, gills, intestine and brain tissues of zebrafish exposed to dissolved MC-LR and MC-RR (0.1-10.0 μgl(-1)). To investigate the time related response of biomarkers, fish were sampled after 4, 7 and 15 days of exposure. Responses varied (i) between MC-LR and MC-RR (for certain groups), (ii) for different enzymes at all time points, and (iii) for different tissues. In general, most of the enzymes followed a bell shaped curve, with an abrupt increase in activity at a particular concentration. It was observed that upon exposure to MC-LR and MC-RR, some enzymes showed an adaptive response after the first time point wherein the enzyme activity increased in some tissues. The increase in enzyme activity is suggestive of their cellular and metabolic adaptations to the continued stress and toxin exposure. Enzyme activities in general increased at lower concentrations (≤ 5.0 μgl(-1)) and decreased at higher concentrations (≥ 5.0 μgl(-1)). An abrupt change in enzyme activities was observed at a particular concentration in all the tissue enzymes. For GPx and GR, there was a differential response in the case of fish exposed to MC-LR and MC-RR, which could be due to the difference in toxicity potentials of these cyanotoxins. In general, initial stress conditions were observed in most of the tissue enzymes following the exposure to microcystins (MCs). This observation suggests that MCs found in trace levels are likely to have deleterious effects on aquatic organisms and can trigger a variety of biochemical responses depending on their specific toxicity.

Determination of aldehydes in rainwater using micro-solid-phase extraction and high-performance liquid chromatography.

A simple and rapid extraction procedure was developed for determining aldehydes in rainwater samples. This extraction technique involved the use of micro-solid-phase extraction in which the sorbent was held within a polypropylene membrane envelope, followed by high-performance liquid chromatographic analysis. Aldehydes such as formaldehyde, acetaldehyde, propionaldehyde and valeraldehyde were used as model compounds. Extraction conditions were optimized. The method linearity ranged between 0.5 and 50 μgl(-1) with the correlation coefficient of 0.987-0.999. The relative standard deviations (RSDs) of the method ranged from 7 to 12%. Method detection limits were in the range of 0.07-0.15 μgl(-1), which is lower than those previously reported for solid-phase microextraction combined with gas chromatography-mass spectrometric techniques. The proposed extraction technique was used for determination of aldehydes in rainwater samples to demonstrate the applicability of the method.

Biosynthesis of gold nanoparticles and related cytotoxicity evaluation using A549 cells.

Biosynthesis of gold nanoparticles (AuNPs) has become an attractive area of research as it is environmentally benign. The toxicity of AuNPs synthesized by chemical routes has been widely studied. However, little is known about the toxicity associated with the biological synthesis of AuNPs. The present study was carried out to synthesize AuNPs using star anise (Illicium verum; a commercially available spice in abundance)and evaluate its toxicity using human epithelial lung cells (A549) in comparison with AuNPs synthesized by the traditional chemical methods (using sodium citrate and sodium borohydride). Apart from cell viability, markers of oxidative stress (reduced glutathione) and cell death (caspases) were also evaluated to understand the mechanisms of toxicity. Cell viability was observed to be 65.7 percent and 72.3 percent in cells exposed to chemically synthesized AuNPs at the highest dose (200nM) as compared to 80.2 percent for biologically synthesized AuNPs. Protective coating/capping of AuNPs by various polyphenolic compounds present in star anise extract appears to be a major contributor to lower toxicity observed in biologically synthesized AuNPs.

Insights into lipidomic perturbations in zebrafish tissues upon exposure to microcystin-LR and microcystin-RR.

This work represents the first study of its kind that was conducted to evaluate changes in lipid metabolic networks following a balneation exposure of adult zebrafish to MCLR (microcystin-leucine-arginine) and MCRR (microcystin-arginine-arginine) at a sublethal dose (10 μg L(-1)) for a period of 30 days. Following the exposure to MCLR and MCRR, gills, liver, intestine, and brain tissues were harvested for metabolite extraction. Extracted metabolites were detected using qTOF-LC-MS (time-of-flight-liquid chromatography-mass spectrometry). Metabolites were identified using Kegg pathways. The identified metabolites are shown on lipid biochemical maps to demonstrate major perturbations in the metabolic machinery. Results showed that most of the metabolic pathways under the lipid class were affected in different tissues of zebrafish following the exposure to MCLR and MCRR (10 μg L(-1) for 30 days). The kind and flux of metabolic perturbations varied among different tissues of the organs after the exposure to MCLR and MCRR with the tissues of gills being the most affected. Among the various lipid pathways, cholesterol synthesis was affected significantly as observed from the highest number of perturbed metabolites in that pathway. Cholesterol is responsible for synthesis of steroid hormones and bile acids, which have been recognized as endocrine signaling molecules. Disruption in the synthesis of these compounds following MCLR/MCRR exposure suggests that MCs are capable of causing endocrine disruption among aquatic organisms even under sublethal conditions. Apart from cholesterol synthesis, various other metabolic pathways belonging to the class of essential fatty acids and lipid oxidation were also observed to be perturbed following a balneation exposure of zebrafish to MCLR/MCRR.

Application of ionic-liquid supported cloud point extraction for the determination of microcystin-leucine-arginine in natural waters.

A cloud point extraction method has been developed using an ionic liquid (1-butyl-3-methylimidazolium hexafluorophosphate, BMiM PF(6)) for the determination of a widely studied cyanotoxin (microcystin leucine-arginine, MCLR) in natural waters. Extraction parameters such as sample pH, extraction temperature, extraction time, the amount of ionic liquid and the amount of extraction volume were investigated and optimized to achieve the maximum extraction efficiency. The results obtained indicated a good linearity with the correlation coefficient of 0.995 over the range of 0.5-50 μg L(-1). The relative standard deviation (RSD) of the method was 7.5% (n=6). The calculated method detection limit was 0.03 μg L(-1) (n=6). The practical applicability of the technique was demonstrated by analyzing water samples (n=9) collected from three different sites in local reservoirs.

Toxicological implications of microcystins for zebrafish embryos in the presence of other environmental pollutants.

Microcystins (MCs) interact with environmental contaminants as well as various other congeners of the MC family in the natural environment and with antioxidants in the exposed organisms. These interactions are likely to modify the toxicological behavior of MCs at the cellular level. The present study was conducted to determine the toxicological response of extracellular MCs in aquatic systems under environmentally relevant conditions. Microcystin-leucine-arginine (MCLR) and microcystin-arginine-arginine (MCRR) were introduced at different concentrations in a single-component (MCLR or MCRR) or dual-component (MCLR and MCRR) system to zebrafish embryos in the presence of inorganic elements (Hg, As, Pb, and Cd) and nutrient species (NO3 (-) , PO4 (3-) , and Cl(-1) ). Hatchability, heart rate, and mortality of zerbrafish embryos were monitored together with changes in the activity of glutathione-S-transferase (GST) to evaluate their response on exposure to MCLR and MCRR. There was a significant reduction in all these parameters at higher doses of MCLR and MCRR (>100 ng/mL), implying bioaccumulation of these MCs in embryos and adverse effects on early development stages of the fish. It was further observed that PO4 (3-) and Cl(-) enhanced the toxic effects of MCLR and MCRR while NO3 (-) attenuated their toxic effects. In contrast, all 4 toxic elements together increased the toxicity of MCLR and MCRR to embryos compared with their single-component counterparts. Thus, the toxic effects of MCs depend not only on their relative environmental concentrations, but also on those of other environmental pollutants and the levels of antioxidants in exposed organisms.

Extracellular Electron Transfer Powers Enterococcus faecalis Biofilm Metabolism

ABSTRACT Enterococci are important human commensals and significant opportunistic pathogens. Biofilm-related enterococcal infections, such as endocarditis, urinary tract infections, wound and surgical site infections, and medical device-associated infections, often become chronic upon the formation of biofilm. The biofilm matrix establishes properties that distinguish this state from free-living bacterial cells and increase tolerance to antimicrobial interventions. The metabolic versatility of the enterococci is reflected in the diversity and complexity of environments and communities in which they thrive. Understanding metabolic factors governing colonization and persistence in different host niches can reveal factors influencing the transition to biofilm pathogenicity. Here, we report a form of iron-dependent metabolism for Enterococcus faecalis where, in the absence of heme, extracellular electron transfer (EET) and increased ATP production augment biofilm growth. We observe alterations in biofilm matrix depth and composition during iron-augmented biofilm growth. We show that the ldh gene encoding l-lactate dehydrogenase is required for iron-augmented energy production and biofilm formation and promotes EET. IMPORTANCE Bacterial metabolic versatility can often influence the outcome of host-pathogen interactions, yet causes of metabolic shifts are difficult to resolve. The bacterial biofilm matrix provides the structural and functional support that distinguishes this state from free-living bacterial cells. Here, we show that the biofilm matrix can immobilize iron, providing access to this growth-promoting resource which is otherwise inaccessible in the planktonic state. Our data show that in the absence of heme, Enterococcus faecalis l-lactate dehydrogenase promotes EET and uses matrix-associated iron to carry out EET. Therefore, the presence of iron within the biofilm matrix leads to enhanced biofilm growth. IMPORTANCE Bacterial metabolic versatility can often influence the outcome of host-pathogen interactions, yet causes of metabolic shifts are difficult to resolve. The bacterial biofilm matrix provides the structural and functional support that distinguishes this state from free-living bacterial cells. Here, we show that the biofilm matrix can immobilize iron, providing access to this growth-promoting resource which is otherwise inaccessible in the planktonic state. Our data show that in the absence of heme, Enterococcus faecalis l-lactate dehydrogenase promotes EET and uses matrix-associated iron to carry out EET. Therefore, the presence of iron within the biofilm matrix leads to enhanced biofilm growth.