François Perreault

Antimicrobial Properties of Graphene Oxide Nanosheets: Why Size Matters

Graphene oxide (GO) is a promising material for the development of antimicrobial surfaces due to its contact-based antimicrobial activity. However, the relationship between GO physicochemical properties and its antimicrobial activity has yet to be elucidated. In this study, we investigated the size-dependency of GO antimicrobial activity using the Gram-negative bacteria Escherichia coli. GO suspensions of average sheet area ranging from 0.01 to 0.65 μm(2) were produced and their antimicrobial activity evaluated in cell suspensions or as a model GO surface coating. The antimicrobial activity of GO surface coatings increased 4-fold when GO sheet area decreased from 0.65 to 0.01 μm(2). The higher antimicrobial effect of smaller GO sheets is attributed to oxidative mechanisms associated with the higher defect density of smaller sheets. In contrast, in suspension assays, GO interacted with bacteria in a cell entrapment mechanism; in this case, the antimicrobial effect of GO increased with increasing sheet area, with apparent complete inactivation observed for the 0.65 μm(2) sheets after a 3 h exposure. However, cell inactivation by GO entrapment was reversible and all initially viable cells could be recovered when separated from GO sheets by sonication. These findings provide useful guidelines for future development of graphene-based antimicrobial surface coatings, where smaller sheet sizes can increase the antimicrobial activity of the material. Our study further emphasizes the importance of an accurate assessment of the antimicrobial effect of nanomaterials when used for antimicrobial surface design.

Polymer coating of copper oxide nanoparticles increases nanoparticles uptake and toxicity in the green alga Chlamydomonas reinhardtii.

Copper oxide nanoparticles (CuO NPs) are frequently used in a polymer-coated form, to be included in paints or fabrics for antimicrobial properties. Their application in antifouling paints may lead to the contamination of aquatic ecosystems. However, the toxicological risk of NPs in the environment is hard to evaluate due to a lack of knowledge on the mechanisms of NP interaction with biological systems. In this study, we investigated the effect of polymer coating on CuO NP toxicity in the green alga Chlamydomonas reinhardtii by comparing bare and polymer-coated CuO NPs prepared from the same CuO nanopowder. Both CuO NP suspensions were toxic to C. reinhardtii after 6 h treatment to concentrations of 0.005-0.04 g L(-1). Bare and polymer-coated CuO NPs induced a decrease of Photosystem II activity and the formation of reactive oxygen species. Polymer-coated CuO NP was found to be more toxic than the uncoated CuO NP. The higher toxicity of CS-CuO NP was mainly associated with the increased capacity of polymer-coated CuO NP to penetrate the cell compared to bare CuO NPs. These results indicates that the high toxicity of polymer-coated CuO NPs in algal cells results of intracellular interactions between NPs and the cellular system.

Evaluation of toxicity and oxidative stress induced by copper oxide nanoparticles in the green alga Chlamydomonas reinhardtii.

Copper oxide nanoparticles (CuO NP) are frequently employed for their antimicrobial properties in antifouling paints. Their extensive use can contaminate aquatic ecosystems. However, the toxicological effects of this NP in the environment are poorly known. In this study, we evaluated the toxicity and oxidative stress induced by CuO NP on Chlamydomonas reinhardtii using several toxicological assays. CuO NP was found to induce growth inhibition and a significant decrease in carotenoids levels. From data on cells density after 72 h of CuO NP exposure in light, the EC50 value was calculated to be 150.45±1.17 mg L(-1) and the NOEC≤100 mg L(-1). Evaluation of esterase activity demonstrates a decrease in cell metabolism activity with the increase of CuO NP concentration. The CuO NP induced an increase of reactive species level (190±0.45% at 1000 mg L(-1) after 72 h of exposition, compared to control) and lipid peroxidation of cellular membranes (73±2% at 1000 mg L(-1) of CuO NP in 72 h of exposition, compared to control). Investigation of CuO NP uptake showed the presence of NP into C. reinhardtii cells in different sites of the cell and, biomarkers of enzymatic antioxidants showed a change of activity after CuO NP exposition. In conclusion, C. reinhardtii was shown to be sensitive to the presence of CuO NP in their environment and CuO NP treatments induced a toxic response from 0.1 mg L(-1) after 72 h of treatment.

Biofouling Mitigation in Forward Osmosis Using Graphene Oxide Functionalized Thin-Film Composite Membranes

Forward osmosis (FO) is an emerging membrane process with potential applications in the treatment of highly fouling feedwaters. However, biofouling, the adhesion of microorganisms to the membrane and the subsequent formation of biofilms, remains a major limitation since antifouling membrane modifications offer limited protection against biofouling. In this study, we evaluated the use of graphene oxide (GO) for biofouling mitigation in FO. GO functionalization of thin-film composite membranes (GO-TFC) increased the surface hydrophilicity and imparted antimicrobial activity to the membrane without altering its transport properties. After 1 h of contact time, deposition and viability of Pseudomonas aeruginosa cells on GO-TFC were reduced by 36% and 30%, respectively, compared to pristine membranes. When GO-TFC membranes were tested for treatment of an artificial secondary wastewater supplemented with P. aeruginosa, membrane biofouling was reduced by 50% after 24 h of operation. This biofouling resistance is attributed to the reduced accumulation of microbial biomass on GO-TFC compared to pristine membranes. In addition, confocal microscopy demonstrated that cells deposited on the membrane surface are inactivated, resulting in a layer of dead cells on GO-TFC that limit biofilm formation. These findings highlight the potential of GO to be used for biofouling mitigation in FO.

Genotoxic effects of copper oxide nanoparticles in Neuro 2A cell cultures.

Copper oxide nanoparticles (CuO NPs) are used for their biocide potential however they were also shown to be highly toxic to mammalian cells. Therefore, the effects of CuO NPs should be carefully investigated to determine the most sensitive processes for CuO NP toxicity. In this study, the genotoxicity of CuO NPs was investigated in vitro, using the mouse neuroblastoma cell line Neuro-2A. Genotoxic effects related to DNA fragmentation, DNA methylation and chromosomal damage, as well as lipid peroxidation, were investigated and compared to cytotoxic effects, measured by the mitochondrial reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide into formazan. Based on mitochondrial activity, CuO NPs were found to be cytotoxic. At the highest concentration tested (400 mg l⁻¹), 63% of cell viability was found in Neuro-2A cells after 24 h of treatment to CuO NPs. CuO NPs were also found to induce DNA fragmentation, lipid peroxidation and micronucleus formation. The micronucleus assay was the most sensitive to evaluate CuO NP genotoxicity and micronucleus frequency was increased significantly at 12.5 mg l⁻¹ CuO NPs after 24h of treatment. At this concentration, no significant change of cell viability was found using the mitochondrial activity assay. These results highlight the important risk of genotoxic effects of CuO NPs and show that genotoxicity assays are a sensitive approach to evaluate the risk of CuO NP toxicity.

Induction to oxidative stress by saxitoxin investigated through lipid peroxidation in Neuro 2A cells and Chlamydomonas reinhardtii alga

Saxitoxin (STX) is a cyanotoxin, which can cause neurotoxic effects and induce ecological changes in aquatic environments, a potential risk to public and environmental health. Many studies of cytotoxicity on animal cells and algae have been performed, although few compare the toxic effects between the two models. In this sense, we investigated the oxidative stress induced by STX (0.4-3.0 nM) in two different cellular models: Neuro-2A (N2A) cells and Chlamydomonas reinhardtii alga by quantification of malondialdehyde (MDA) levels as indicative of lipid peroxidation (LPO). Also was evaluated the antioxidant defense of these cells systems after exposure to STX by the addition of antioxidants in N2A cells culture, and by the measure of antioxidants enzymes activity in C. reinhardtii cells. The MDA levels of N2A cells increased from 15% to 113% for 0.4 and 3.0 nM of STX, respectively, as compared to control. Superoxide-dismutase and catalase did not appear to protect the cell from STX effect while, in cells treated with vitamin E, the rates of MDA production decreased significantly, except for higher concentrations of STX. No MDA productions were observed in algal cells however some effects on antioxidant enzymes activity were observed when algae were exposed to 3.0 nM STX. Our results indicate that the concentrations of STX that may induce oxidative stress through LPO are different in animal and phytoplankton communities. A combination of algal and animal bioassays should be conducted for reliable assessment of oxidative stress induced by STX.

Shape-Dependent Surface Reactivity and Antimicrobial Activity of Nano-Cupric Oxide

Shape of engineered nanomaterials (ENMs) can be used as a design handle to achieve controlled manipulation of physicochemical properties. This tailored material property approach necessitates the establishment of relationships between specific ENM properties that result from such manipulations (e.g., surface area, reactivity, or charge) and the observed trend in behavior, from both a functional performance and hazard perspective. In this study, these structure-property-function (SPF) and structure-property-hazard (SPH) relationships are established for nano-cupric oxide (n-CuO) as a function of shape, including nanospheres and nanosheets. In addition to comparing these shapes at the nanoscale, bulk CuO is studied to compare across length scales. The results from comprehensive material characterization revealed correlations between CuO surface reactivity and bacterial toxicity with CuO nanosheets having the highest surface reactivity, electrochemical activity, and antimicrobial activity. While less active than the nanosheets, CuO nanoparticles (sphere-like shape) demonstrated enhanced reactivity compared to the bulk CuO. This is in agreement with previous studies investigating differences across length-scales. To elucidate the underlying mechanisms of action to further explain the shape-dependent behavior, kinetic models applied to the toxicity data. In addition to revealing different CuO material kinetics, trends in observed response cannot be explained by surface area alone. The compiled results contribute to further elucidate pathways toward controlled design of ENMs.

Dichromate effect on energy dissipation of photosystem II and photosystem I in Chlamydomonas reinhardtii

In this study, we investigated the energy dissipation processes via photosystem II and photosystem I activity in green alga Chlamydomonas reinhardtii exposed to dichromate inhibitory effect. Quantum yield of photosystem II and also photosystem I were highly decreased by dichromate effect. Such inhibition by dichromate induced strong quenching effect on rapid OJIP fluorescence transients, indicating deterioration of photosystem II electron transport via plastoquinone pool toward photosystem I. The decrease of energy dissipation dependent on electron transport of photosystem II and photosystem I by dichromate effect was associated with strong increase of non-photochemical energy dissipation processes. By showing strong effect of dichromate on acceptor side of photosystem I, we indicated that dichromate inhibitory effect was not associated only with PSII electron transport. Here, we found that energy dissipation via photosystem I was limited by its electron acceptor side. By the analysis of P700 oxido-reduction state with methylviolagen as an exogenous PSI electron transport mediator, we showed that PSI electron transport discrepancy induced by dichromate effect was also caused by inhibitory effect located beyond photosystem I. Therefore, these results demonstrated that dichromate has different sites of inhibition which are associated with photosystem II, photosystem I and electron transport sink beyond photosystems.

Investigation of animal and algal bioassays for reliable saxitoxin ecotoxicity and cytotoxicity risk evaluation

Contamination of water bodies by saxitoxin can result in various toxic effects in aquatic organisms. Saxitoxin contamination has also been shown to be a threat to human health in several reported cases, even resulting in death. In this study, we evaluated the sensitivity of animal (Neuro-2A) and algal (Chlamydomonas reinhardtii) bioassays to saxitoxin effect. Neuro-2A cells were found to be sensitive to saxitoxin, as shown by a 24 h EC50 value of 1.5 nM, which was obtained using a cell viability assay. Conversely, no saxitoxin effect was found in any of the algal biomarkers evaluated, for the concentration range tested (2-128 nM). These results indicate that saxitoxin may induce toxic effects in animal and human populations at concentrations where phytoplankton communities are not affected. Therefore, when evaluating STX risk of toxicity, algal bioassays do not appear to be reliable indicators and should always be conducted in combination with animal bioassays.

Impaired Performance of Pressure-Retarded Osmosis due to Irreversible Biofouling.

Next-generation pressure-retarded osmosis (PRO) approaches aim to harness the energy potential of streams with high salinity differences, such as wastewater effluent and seawater desalination plant brine. In this study, we evaluated biofouling propensity in PRO. Bench-scale experiments were carried out for 24 h using a model wastewater effluent feed solution and simulated seawater desalination brine pressurized to 24 bar. For biofouling tests, wastewater effluent was inoculated with Pseudomonas aeruginosa and artificial seawater desalination plant brine draw solution was seeded with Pseudoalteromonas atlantica. Our results indicate that biological growth in the feed wastewater stream channel severely fouled both the membrane support layer and feed spacer, resulting in ∼50% water flux decline. We also observed an increase in the pumping pressure required to force water through the spacer-filled feed channel, with pressure drop increasing from 6.4±0.8 bar m(-1) to 15.1±2.6 bar m(-1) due to spacer blockage from the developing biofilm. Neither the water flux decline nor the increased pressure drop in the feed channel could be reversed using a pressure-aided osmotic backwash. In contrast, biofouling in the seawater brine draw channel was negligible. Overall, the reduced performance due to water flux decline and increased pumping energy requirements from spacer blockage highlight the serious challenges of using high fouling potential feed sources in PRO, such as secondary wastewater effluent. We conclude that PRO power generation using wastewater effluent and seawater desalination plant brine may become possible only with rigorous pretreatment or new spacer and membrane designs.