M. Bhuvaneshwari

Combined toxicity of two crystalline phases (anatase and rutile) of Titania nanoparticles towards freshwater microalgae: Chlorella sp.

In view of the increasing usage of anatase and rutile crystalline phases of titania NPs in the consumer products, their entry into the aquatic environment may pose a serious risk to the ecosystem. In the present study, the possible toxic impact of anatase and rutile nanoparticles (individually and in binary mixture) was investigated using freshwater microalgae, Chlorella sp. at low exposure concentrations (0.25, 0.5 and 1mg/L) in freshwater medium under UV irradiation. Reduction of cell viability as well as a reduction in chlorophyll content were observed due to the presence of NPs. An antagonistic effect was noted at certain concentrations of binary mixture such as (0.25, 0.25), (0.25, 0.5), and (0.5, 0.5) mg/L, and an additive effect for the other combinations, (0.25, 1), (0.5, 0.25), (0.5, 1), (1, 0.25), (1, 0.5), and (1, 1) mg/L. The hydrodynamic size analyses in the test medium revealed that rutile NPs were more stable in lake water than the anatase and binary mixtures [at 6h, the sizes of anatase (1mg/L), rutile NPs (1mg/L), and binary mixture (1, 1mg/L) were 948.83±35.01nm, 555.74±19.93nm, and 1620.24±237.87nm, respectively]. The generation of oxidative stress was found to be strongly dependent on the crystallinity of the nanoparticles. The transmission electron microscopic images revealed damages in the nucleus and cell membrane of algal cells due to the interaction of anatase NPs, whereas rutile NPs were found to cause chloroplast and internal organelle damages. Mis-shaped chloroplasts, lack of nucleus, and starch-pyrenoid complex were noted in binary-treated cells. The findings from the current study may facilitate the environmental risk assessment of titania NPs in an aquatic ecosystem.

Cytotoxicity of ZnO NPs towards fresh water algae Scenedesmus obliquus at low exposure concentrations in UV-C, visible and dark conditions

Continuous increase in the usage of ZnO nanoparticles in commercial products has exacerbated the risk of release of these particles into the aquatic environment with possible harmful effects on the biota. In the current study, cytotoxic effects of two types of ZnO nanoparticles, having different initial effective diameters in filtered and sterilized lake water medium [487.5±2.55 nm for ZnO-1 NPs and 616.2±38.5 nm for ZnO-2 NPs] were evaluated towards a dominant freshwater algal isolate Scenedesmus obliquus in UV-C, visible and dark conditions at three exposure concentrations: 0.25, 0.5 and 1 mg/L. The toxic effects were found to be strongly dependent on the initial hydrodynamic particle size in the medium, the exposure concentrations and the irradiation conditions. The loss in viability, LDH release and ROS generation were significantly enhanced in the case of the smaller sized ZnO-1 NPs than in the case of ZnO-2 NPs under comparable test conditions. The toxicity of both types of ZnO NPs was considerably elevated under UV-C irradiation in comparison to that in dark and visible light conditions, the effects being more enhanced in case of ZnO-1 NPs. The size dependent dissolution of the ZnO NPs in the test medium and possible toxicity due to the released Zn(2+) ions was also noted. The surface adsorption of the nanoparticles was substantiated by scanning electron microscopy. The internalization/uptake of the NPs by the algal cells was confirmed by fluorescence microscopy, transmission electron microscopy, and elemental analyses.

Different modes of TiO2 uptake by Ceriodaphnia dubia: relevance to toxicity and bioaccumulation.

The extensive environmental exposure of engineered metal oxide nanoparticles (NPs) may result in their bioaccumulation in aquatic organisms leading to their biotransfer in a food chain through various routes in a freshwater ecosystem. The present study focuses on the possible modes of TiO2 NP trophic transfer to Ceriodaphnia dubia, in presence and/absence of its diet, Scenedesmus obliquus (primary producer). The acute exposure studies (48h) were designed to have daphnids exposed to (i) the free NPs, (ii) both the free and the algae-borne NPs; and (iii) only the algae-borne NPs in separate tests to understand the possible routes of NP transfer. The dietary uptake of TiO2 NPs (algae-borne) was found to be the primary route for NP biotransfer with ∼70% of total NP uptake. Interestingly, in a separate study it was noticed that the NPs coated with algal exudates were easily taken up by daphnids as compared to pristine NPs of same concentrations, leading to their higher bioaccumulation. A chronic toxicity study, where daphnids were exposed to both free and algae-borne NPs for 21 days was undertaken to comprehend the TiO2 NP effect on daphnia growth and reproduction upon chronic exposure and also the bioaccumulation potential. Both acute and chronic exposure studies suggested higher bioaccumulation of TiO2 in daphnids when the particles were less toxic to the diet (algae).

Toxic effect of Cr(VI) in presence of n-TiO2 and n-Al2O3 particles towards freshwater microalgae.

The reactivity and toxicity of the soluble toxicants in the presence of the engineered nanomaterials is not well explored. In this study, the probable effects of TiO2 and Al2O3 nanoparticles (n-TiO2, n-Al2O3) on the toxicity of Cr(VI) were assessed with the dominant freshwater algae, Scenedesmus obliquus, in a low range of exposure concentrations (0.05, 0.5 and 1μg/mL). In the presence of 0.05μg/mL n-TiO2, the toxicity of Cr(VI) decreased considerably, which was presumably due to the Cr(VI) adsorption on the nanoparticle surface leading to its aggregation and precipitation. The elevated n-TiO2 concentrations (0.5 and 1μg/mL) did not significantly influence Cr(VI) bio-availability, and a dose dependent toxicity of Cr(VI) was observed. On the other hand, n-Al2O3 did not have any significant effect on the Cr(VI) toxicity. The microscopic observations presented additional information on the morphological changes of the algal cells in the presence of the binary toxicants. The generation of reactive oxygen species (ROS) suggested contribution of oxidative stress on toxicity and LDH release confirmed membrane permeability of algal cells upon stress.

Acute toxicity and accumulation of ZnO NPs in Ceriodaphnia dubia: Relative contributions of dissolved ions and particles.

Although the ecotoxicological effects of various metal oxide nanoparticles on aquatic organisms are being actively studied, the contributions of particles and dissolved ions towards toxicity are still not well understood. The current study aims to assess the contribution of ZnO NP(particle) and ZnO NP(ion) to the overall toxicity and accumulation of ZnO NP(total) in Ceriodaphnia dubia. The aggregation and dissolution kinetics were studied for three different sizes (50nm, 100nm and bulk) of ZnO particles at 0.05, 0.12, 0.25 and 0.5mg/L concentrations in the sterile lake water medium at 6, 12, 24, and 48h intervals. The 48h LC50 of ZnO NP(total) was found to be 0.431, 0.605 and 0.701mg/L for 50, 100nm and bulk particles exposure. However, LC50 of Zn(ion) was found to be 1.048, 1.343 and 2.046mg/L for dissolved ions from different sizes (50nm, 100nm, and bulk) of ZnO particles. At LC50 concentration, the accumulation of 90-95% was noted for the NP(particles) across the sizes employed, while only about 4-5% contribution was from the NP(ion) to the overall accumulation NP(total). The relative contribution of ZnO NP(ion) to overall toxicity and accumulation was found to be lesser than that of ZnO NP(particles) across the sizes used in the study.

Toxicity and accumulation of Copper oxide (CuO) nanoparticles in different life stages of Artemia salina

Metal nanoparticles production rate and its applications have raised concerns about their release and toxicity to the aquatic and terrestrial organisms. The primary size of Copper Oxide nanoparticles (CuO NPs) was found to be 114±36nm using Scanning Electron Microscopy (SEM) and a significant increase in the hydrodynamic diameter of CuO NP was seen within 1h of interaction. The median lethal concentration (LC50) values obtained from the acute toxicity studies on different life stages of Artemia salina was found to be 61.4, 35, 12.2 and 175.2mg/L for 1d, 2d, 7d old and adult, respectively. The toxicity associated changes in biochemical markers such as Catalase, Reduced glutathione and Glutathione-S-Transferase were evident. The accumulation of Cu nanoparticles into the gut of Artemia salina was the major reason for toxicity. This study demonstrate the toxicity of CuO NPs to Artemia salina, and the obtained results necessitate the detailed investigation on the possible eco-toxicological implication of these nanomaterials.

Differential effects of P25 TiO2 nanoparticles on freshwater green microalgae: Chlorella and Scenedesmus species.

P25 TiO2 nanoparticles majorly used in cosmetic products have well known detrimental effects towards the aquatic environment. In a freshwater ecosystem, Chlorella and Scenedesmus are among the most commonly found algal species frequently used to study the effects of metal oxide nanoparticles. A comparative study has been conducted herein to investigate differences in the toxic effects caused by these nanoparticles towards the two algae species. The three different concentrations of P25 TiO2 NPs (0.01, 0.1 & 1μg/mL, i.e., 0.12, 1.25 and 12.52μM) were selected to correlate surface water concentrations of the nanoparticles, and filtered and sterilized fresh water medium was used throughout this study. There was significant increase (p<0.001) in hydrodynamic diameter of nanoparticles with respect to both, time (0, 24, 48 and 72h) as well as concentration under all the exposure conditions. Although, significant dose-dependent morphological (surface area & biovolume) interspecies variations were not observed, it was evident at the highest concentration of exposure within individuals. At 1μg/mL exposure concentration, a significant difference in toxicity was noted between Chlorella and Scenedesmus under only visible light (p<0.001) and UVA (p<0.01) irradiation conditions. The viability data were well supported by the results obtained for oxidative stress induced by NPs on the cells. At the highest exposure concentration, superoxide dismutase and reduced glutathione activities were assessed for both the algae under all the irradiation conditions. Increased catalase activity and LPO release complemented the cytotoxic effects observed. Significant interspecies variations were noted for these parameters under UVA and visible light exposed cells of Chlorella and Scenedesmus species, which could easily be correlated with the uptake of the NPs.

Individual and binary toxicity of anatase and rutile nanoparticles towards Ceriodaphnia dubia

Increasing usage of engineered nanoparticles, especially Titanium dioxide (TiO2) in various commercial products has necessitated their toxicity evaluation and risk assessment, especially in the aquatic ecosystem. In the present study, a comprehensive toxicity assessment of anatase and rutile NPs (individual as well as a binary mixture) has been carried out in a freshwater matrix on Ceriodaphnia dubia under different irradiation conditions viz., visible and UV-A. Anatase and rutile NPs produced an LC50 of about 37.04 and 48mg/L, respectively, under visible irradiation. However, lesser LC50 values of about 22.56 (anatase) and 23.76 (rutile) mg/L were noted under UV-A irradiation. A toxic unit (TU) approach was followed to determine the concentrations of binary mixtures of anatase and rutile. The binary mixture resulted in an antagonistic and additive effect under visible and UV-A irradiation, respectively. Among the two different modeling approaches used in the study, Marking-Dawson model was noted to be a more appropriate model than Abbott model for the toxicity evaluation of binary mixtures. The agglomeration of NPs played a significant role in the induction of antagonistic and additive effects by the mixture based on the irradiation applied. TEM and zeta potential analysis confirmed the surface interactions between anatase and rutile NPs in the mixture. Maximum uptake was noticed at 0.25 total TU of the binary mixture under visible irradiation and 1 TU of anatase NPs for UV-A irradiation. Individual NPs showed highest uptake under UV-A than visible irradiation. In contrast, binary mixture showed a difference in the uptake pattern based on the type of irradiation exposed.

Antibacterial and antifouling activities of chitosan/TiO2/Ag NPs nanocomposite films against packaged drinking water bacterial isolates.

TiO2 and Ag NPs are widely used as antibacterial agents against many bacterial pathogens. Chitosan (polymer) itself acts as a strong antibacterial agent. Hence, chitosan/TiO2/Ag NPs incorporated nanocomposite film was prepared against packed drinking water bacterial strains. A concentration-dependent increase in the reduction of cell viability was observed in all the isolates under UV-C and dark exposure conditions. The bacteria consortium showed greater resistance against antibacterial effects of chitosan/TiO2/Ag nanocomposite as compared to single isolates. Glycocalyx test and mass assessment conclude the effective antibacterial activity by inhibiting bacterial adhesion on the film surface. The release of LDH and generation of ROS act as the predominant antibacterial mechanism induced by TiO2/Ag NPs. Surface characterization of chitosan/TiO2/Ag nanocomposite was studied by FTIR and XRD analyses and SEM analysis after interaction with the bacteria.

Toxicity, accumulation, and trophic transfer of chemically and biologically synthesized nano zero valent iron in a two species freshwater food chain.

The impact of bio-remediation agent nZVI on environment is still inadequately understood, especially on aquatic food web. The study presented here has therefore considered both chemical (CS) and biological (BS) synthetic origins of nZVI and their effects on both algae and daphnia. The study is unique in its attempt to explore the possibility of trophic transfer from algae to its immediate higher niche (daphnia as the model). An equal weightage of the effects of both CS and BS nZVI on algae and daphnia has been explored here; hence it allows us to compare the capping of nZVI on toxicity. To examine the causes of observed lethality- ROS generation, effects on the activity of oxidative enzymes, membrane damage and biouptake of nZVI was analysed. The overall outcome of CS and BS nZVI on lethality was significantly different in algae and daphnia, where daphnia demonstrated relatively higher sensitivity against CS nZVI. Algae demonstrated considerable differences in CS and BS nZVI toxicity only at higher concentration. This study did not show a probable biomagnification and trophic transfer from algae to daphnia under the experimental conditions even at the highest exposure concentration. The study instigates the importance of trophic transfer to understand the possible biomagnification of nZVI among organisms of different trophic levels and eventually the consequences on environment.