Article

Effects of carbon and silicon nanotubes and carbon nanofibers on marine microalgae Heterosigma akashiwo

Authors:
  • Siberian Federal Scientific Centre of Agrobiotechnology
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Abstract

The effect of carbon and silicon nanotubes (CNTs and SiNTs) and carbon nanofibers (CNFs) to microscopic marine algae Heterosigma akashiwo was studied, using algal growth inhibition for 3 days (acute effect) and 7 days (chronic effect) as toxicity endpoints. The criterion of the toxic effect was the statistically significant reduction of the number of algal cells in the exposed samples compared to the control. Samples did not demonstrate toxic effects at doses 1 mg/l and 10 mg/l. CNTs and SiNTs samples at 100 mg/l exhibited both acute and chronic toxic effects. We assume that the main cause of cell death in these samples was related to the mechanical damage of cell integrity. CNFs at concentrations of 100 mg/l did not inhibit algal growth, but cells with irregular shapes were observed, which were not observed after exposure to CNTs and SiNTs. Nickel impurities present in CNFs samples are presumably the main cause of observed cell deformations.

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... The entrance of synthetic nanofibers, NPs, and NMs to water bodies has been reported in earlier studies [36][37][38][39]. Aquatic organisms such as microalgae species are known as reliable research objects in toxicology, and they are one of the most commonly used organisms in aquatic toxicity assessment of NPs [40][41][42][43]. Previous research has established that carbon nanotubes could cause changes in biochemical composition of microalgae [44], could attach to or penetrate algal cells [45], and the NPs may become involved in the transfer between the levels of food chain [46]. ...
... Previous research has established that carbon nanotubes could cause changes in biochemical composition of microalgae [44], could attach to or penetrate algal cells [45], and the NPs may become involved in the transfer between the levels of food chain [46]. In contrast to carbon nanotubes, there is much less information about effects of carbon nanofibers and silica nanotubes on microalgae [41]. ...
... Carbon nanotubes and nanofibers were synthesized and characterized in the Boreskov Institute of Catalysis (Novosibirsk, Russia) [47]. The structural features of carbon NPs ware assessed by Raman spectroscopy in our earlier report [41]. The length of carbon nanotubes was hundreds of times larger than the diameter and in water suspension; the particles could cohere into the spheres up to tens of micrometers in diameter. ...
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... MWCNTs exert toxic effects on cellular metabolic activity, cell components, and plasma membrane of lamentous green microalgae (Munk et al., 2017). Marine algae Heterosigma akashiwo had no toxic effects at CNTs doses of 1 and 10 mg·L − 1 , while 100 mg·L − 1 exerted both acute and chronic toxic effects (Pikula et al., 2018). It has been reported that the CNTs level higher than 40 mg·L − 1 could be fatal to freshwater algae within a few days, such as Chlorella vulgaris (Long et al., 2012;Sohn et al., 2015). ...
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... Results revealed dosedependent increase in lipid peroxidation due to reactive oxygen species generation. However, Pikula et al. (2018) reported that the detrimental effects on the algae Heterosigma akashiwo due to carbon and silicon nanotubes and carbon nanofibers (CNFs) were related to the mechanical damage rather than toxicity. ...
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... We used the VEPs at three different concentrations i.e. 1, 10, and 100 mg/l in a tetra-replicate experiment. This dosage selection is provided by the data of previous studies where the toxicity of particulate matter was assessed by the tests with microalgae or other aquatic organisms (Durga et al., 2014;Correa et al., 2016Correa et al., , 2017Pikula et al., 2018). ...
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Carbon nanofibers (CNFs) have initiated a new era of outstanding biological applications for future generations. CNFs can be used as efficient and unique biocompatible reinforcing materials, biosensors, perfect platforms for supporting nanoparticles and immobilizing biomolecules, and wound dressing support due to their surface functionality, ensuring biocompatibility and coupling with metallic nanoparticles or organics. Using CNFs helps to build artificial joints, promoting tissue, organ, and nerve regeneration and development. In addition, multiple published studies have revealed CNF's ability to boost the impact and chemosensitization of anticancer therapies for a range of tumor types while lowering undesirable drug effects. Also, the toxicity of CNFs as effective biomaterials was explored, as well as the limits. CNF research focuses on biomedical applications, such as tissue engineering and biosensors. The bulk of these biological applications utilise CNTs, graphene, and fullerene, although CNFs are still rarely cited in these domains. As a result, increased emphasis and effort should be placed on examining the potential benefits of adopting CNFs. Lastly, the dearth of proven evidence on CNF toxicity should push scientists to complete the dataset necessary for CNF risk assessment. For example, short- and long-term chronic animal bioassays and standard tests can be utilized to replicate real-world conditions for occupational, local, and systemic administration of CNFs.
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Nanoparticles (NPs) are widely used for their special physical properties and released into the natural environment. When two types of NPs exist in the same environment, the presence of one type of NP may affect the properties of the other type of NP. This study investigated the toxic effects of multi-walled carbon nanotubes (MWCNTs) and copper oxide nanoparticles (CuO NPs) on Tetradesmus obliquus. Both NPs had toxic effects on algae, and the toxic effects of MWCNTs were significantly stronger than CuO NPs which the 96-hr median effective concentration to algae were 33.8 and 169.2 mg/L, respectively. Oxidative stress and cell membrane damage were the main reasons for the toxicity of NPs to algae, and they were concentration-dependent, and the existence of CuO NPs in some groups reduced cell membrane damage caused by MWCNTs which may because that CuO NPs formed heteroaggregation with MWCNTs, reducing the contact of nanoparticles with cell membranes, then reducing physical damage. Scanning electron microscopy (SEM) and transmission electron microscope (TEM) results indicated cell damage, the heteroaggregation of MWCNTs-CuO NPs and obvious nanoparticles internalization. In some groups, the presence of CuO NPs significantly reduced reactive oxygen species (ROS) level induced by MWCNTs. However, for the highest concentration group, the ROS level was much higher than that of the two NPs alone treatment groups, which might be related to the high concentration of MWCNTs promoting the internalization of CuO NPs. MWCNTs and CuO NPs affected and interacted with each other, causing more complex toxic effects on aquatic organisms.
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Great attention should be paid to the possible impacts of various nanoparticles on aquatic organism. Freshwater diatoms are essential components of phytoplankton and play a critical role in bioassessment of nanoparticle exposure in the environment. In this study, cell growth was inhibited by TiO2-NPs and MWCNTs, the 24 h EC50 values of TiO2-NPs and MWCNTs to Nitzschia frustulum were 20.75 and 24.64 mg L⁻¹, respectively. Significant decreases of Chl a content after TiO2-NPs exposures were detected and the Chl a content of N. frustulum was obviously increased by MWCNTs treatment at lower concentration. The ROS was detected in N. frustulum after TiO2-NPs and MWCNTs exposures. The MDA content was significantly induced by TiO2-NPs at lower concentrations of 24 and 48 h exposure; meanwhile, it increased at all tested concentrations at 24 h MWCNTs exposure. The SOD enzyme was induced by 72 and 96 h TiO2-NPs exposure, and increased by MWCNTs treatment at 96 h in N. frustulum. Scanning electron microscopy results revealed that N. frustulum had obvious cell deformation after TiO2-NPs treatment. The result showed that the physiological and biochemical response mechanisms after NPs exposure of diatom were species-specific, and in relation to the exposure concentration and time. © 2018
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Extracellular matrix (ECM) is an extraordinarily complex and unique meshwork composed of structural proteins and glycosaminoglycans. The ECM provides essential physical scaffolding for the cellular constituents, as well as contributes to crucial biochemical signaling. Importantly, ECM is an indispensable part of all biological barriers and substantially modulates the interchange of the nanotechnology products through these barriers. The interactions of the ECM with nanoparticles (NPs) depend on the morphological characteristics of intercellular matrix and on the physical characteristics of the NPs and may be either deleterious or beneficial. Importantly, an altered expression of ECM molecules ultimately affects all biological processes including inflammation. This review critically discusses the specific behavior of NPs that are within the ECM domain, and passing through the biological barriers. Furthermore, regenerative and toxicological aspects of nanomaterials are debated in terms of the immune cells-NPs interactions.
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The marine bivalves, mussels Crenomytilusgrayanus (Dunker, 1853) and scallops Swiftopecten swifti (Bernardi, 1858), were in vivo exposed to 12-14 nm multi-walled carbon nanotubes (MWNTs) for up to 48 h. Microscopic analysis in combination with the RAMAN spectrophotometry revealed the MWNT aggregates on the gills surface and inside the gut of all exposed individuals. After 48 h exposure, there were no changes in the total cell count, the average cell size and granularity in the hemolymph of mussels, while in the scallops the total hemocyte count was significantly reduced, and the average hemocyte granularity increased. Biochemical markers of oxidative stress (activity of glutathione-S-transferase and catalase, concentration of reduced glutathione, and the degree of lipid peroxidation) did not change significantly in the digestive gland of both mussels and scallops. In hemolymph, catalase activity increased as compared to control in both mussels and scallops. Moreover, concentration of reduced glutathione increased in hemolymph of scallops on the second day of exposure to MWNTs. The data obtained indicate that MWNTs may affect different bivalve mollusks more or less strongly under the same exposure conditions.
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With the expansion of the nanomedicine field, the knowledge focusing on the behavior of nanoparticles in the biological milieu has rapidly escalated. Upon introduction to a complex biological system, nanomaterials dynamically interact with all the encountered biomolecules and form the protein "bio-corona." The decoration with these surface biomolecules endows nanoparticles with new properties. The present review will address updates of the protein bio-corona characteristics as influenced by nanoparticle's physicochemical properties and by the particularities of the encountered biological milieu. Undeniably, bio-corona generation influences the efficacy of the nanodrug and guides the actions of innate and adaptive immunity. Exploiting the dynamic process of protein bio-corona development in combination with the new engineered horizons of drugs linked to nanoparticles could lead to innovative functional nanotherapies. Therefore, bio-medical nanotechnologies should focus on the interactions of nanoparticles with the immune system for both safety and efficacy reasons.
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Studies of the neurobehavioral effects of carbon nanomaterials, particularly those of multi-walled carbon nanotubes (MWCNTs), have concentrated on cognitive effects, but data are scarce. The aim of this study was to assess the influence of MWCNTs on a number of higher nervous system functions of Wistar rats. For a period of 10 days, two experimental groups were fed with MWCNTs of different diameters (MWCNT-1 group, 8-10 nm; MWCNT-2 group, 18-20 nm) once a day at a dosage of 500 mg/kg. In the open-field test, reductions of integral indications of researching activity were observed for the two MWCNT-treated groups, with a parallel significant (P<0.01) increase in stress levels for these groups compared with the untreated control group. In the elevated plus-maze test, integral indices of researching activity in the MWCNT-1 and MWCNT-2 groups reduced by day 10 by 51 and 62%, respectively, while rat stress levels remained relatively unchanged. In the universal problem solving box test, reductions in motivation and energy indices of researching activity were observed in the two experimental groups. Searching activity in the MWCNT-1 group by day 3 was reduced by 50% (P<0.01) and in the MWCNT-2 group the relevant reduction reached 11.2%. By day 10, the reduction compared with controls, was 64% (P<0.01) and 58% (P<0.01) for the MWCNT-1 and MWCNT-2 groups, respectively. In conclusion, a series of specific tests demonstrated that MWCNT-treated rats experienced a significant reduction of some of their cognitive abilities, a disturbing and worrying finding, taking into consideration the continuing and accelerating use of carbon nanotubes in medicine and science.
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Abstract The impact of two types of multi-walled carbon nanotubes (MWCNTs) (12-14 nm) with different content of metallic impurities (purified and unpurified nanotubes) on peroxidation processes, the status of immune cells in healthy volunteers and gene expression combined to pathway analysis was studied in vitro. From the study it was shown that the main mechanism of action for both types of MWCNTs is induction of oxidative stress, the intensity of which is directly related to the amount of metallic impurities. Unpurified MWCNTs produced twice as high levels of oxidation than the purified CNTs inducing thus more intense mitochondrial dysfunction. All the above were also verified by gene expression analysis of 2 different human cellular cultures (lung epithelium and keratinoma cells) and the respective pathway analysis; modulation of genes activating the NFkB pathway is associated to inflammatory responses. This may cause a perturbation in the IL-6 signaling pathway in order to regulate inflammatory processes and compensate for apoptotic changes. A plausible hypothesis for the immunological effects observed in vivo, are considered as the result of the synergistic effect of systemic (mediated by cells of the routes of expjsure) and local inflammation (blood cells).
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Microalgae are a source of numerous compounds that can be used in many branches of industry. Synthesis of such compounds in microalgal cells can be amplified under stress conditions. Exposure to various metals can be one of methods applied to induce cell stress and synthesis of target products in microalgae cultures. In this review, the potential of producing diverse biocompounds (pigments, lipids, exopolymers, peptides, phytohormones, arsenoorganics, nanoparticles) from microalgae cultures upon exposure to various metals, is evaluated. Additionally, different methods to alter microalgae response towards metals and metal stress are described. Finally, possibilities to sustain high growth rates and productivity of microalgal cultures in the presence of metals are discussed.
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The increasing production and use of engineered nanomaterials (ENMs) inevitably results in their higher concentrations in the environment. This may lead to undesirable environmental effects and thus warrants risk assessment. The ecotoxicity testing of a wide variety of ENMs rapidly evolving in the market is costly but also ethically questionable when bioassays with vertebrates are conducted. Therefore, alternative methods, e.g., models for predicting toxicity mechanisms of ENMs based on their physico-chemical properties (e.g., quantitative (nano)structure-activity relationships, QSARs/QNARs), should be developed. While the development of such models relies on good-quality experimental toxicity data, most of the available data in the literature even for the same test species are highly variable. In order to map and analyse the state of the art of the existing nanoecotoxicological information suitable for QNARs, we created a database NanoE-Tox that is available as Supporting Information File 2. The database is based on existing literature on ecotoxicology of eight ENMs with different chemical composition: carbon nanotubes (CNTs), fullerenes, silver (Ag), titanium dioxide (TiO2), zinc oxide (ZnO), cerium dioxide (CeO2), copper oxide (CuO), and iron oxide (FeOx; Fe2O3, Fe3O4). Altogether, NanoE-Tox database consolidates data from 224 articles and lists altogether 1,518 toxicity values (EC50/LC50/NOEC) with corresponding test conditions and physico-chemical parameters of the ENMs as well as reported toxicity mechanisms and uptake of ENMs in the organisms. 35% of the data in NanoE-Tox concerns ecotoxicity of Ag NPs, followed by TiO2 (22%), CeO2 (13%), and ZnO (10%). Most of the data originates from studies with crustaceans (26%), bacteria (17%), fish (13%), and algae (11%). Based on the median toxicity values of the most sensitive organism (data derived from three or more articles) the toxicity order was as follows: Ag > ZnO > CuO > CeO2 > CNTs > TiO2 > FeOx. We believe NanoE-Tox database contains valuable information for ENM environmental hazard estimation and development of models for predicting toxic potential of ENMs
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A survey was conducted for production of the reactive oxygen species superoxide by 37 species (65 strains) of microalgae including dinoflagellates, raphidophytes, chlorophytes, prasinophytes, eustigmatophytes and prymnesiophytes. Ichthyotoxic raphidophyte species of Chattonella were found to produce the highest environmental levels of superoxide (177×104 total chemiluminescence units). However, ichthyotoxic dinoflagellates (Karenia, Alexandrium) and the prymnesiophyte Prymnesium were also found to produce significant levels of superoxide (4×104, 3×104 and 5×104 chemiluminescence units, respectively), equivalent to that of other raphidophyte species of Heterosigma and Fibrocapsa (6×104 and 2×104, respectively). A direct relationship between cell size and superoxide production was observed (r 2 = 0.94), with larger cells producing more superoxide per cell. Chattonella produced the most superoxide per cell (expressed as cellular chemiluminescence units), followed by the dinoflagellate species Karenia, Alexandrium, Takayama and Gymnodinium. Small cells, such as the raphidophyte Heterosigma and the prymnesiophyte Prymnesium produced very little superoxide per cell (cellular chemiluminescence units), but potentially could still produce high total levels of superoxide if present at high biomass levels. Species commonly used as aquaculture bivalve feeds such as Dunaliella, Tetraselmis, Nannochloropsis and Pavlova produced negligible levels of superoxide, even at high biomass. We speculate that superoxide, while not the sole ichthyotoxic principle, may play a wider role in algal toxicity than previously considered, and propose a broad classification of microalgae based upon superoxide production.
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The reaction of bivalves Modiolus modiolus to pulse (for 24 and 48 h) exposure with multiwalled carbon nanotubes (MWCNTs) (12–14 nm, MWNT concentration in sea water of 100 mg/L) is manifested in the ingestion of MWCNT aggregates formed in seawater despite their rapid sedimentation from the water column to the bottom of the aquariums. After 24 h, the MWCNT aggregates are observed in the intestinal lumen (size of 10 to 150 μm) and in the tubules of the digestive gland (10 to 50 μm). After 48 h, only large aggregates in contact with mucus and desquamated epithelium fragments are detected in the lumen of the intestine. The smallest aggregates seem to be inside epithelial cells. In the intestine, digestive gland, and gills, MWCNT aggregates induce histopathological changes in the epithelium (erosion, necrosis, trend towards increased vacuolization of the cells) and swelling of the connective tissue. In the gill epithelium after 48 h, patterns morphologically corresponding to apoptosis are observed. Despite significant organ damage, no change in the cellular composition of the hemolymph in mussels exposed to the MWCNTs is found.
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Carbon nanotubes (CNTs) and graphene have built broad interest in most areas of science and engineering because of their extraordinary physical, mechanical, thermal and optical properties. Graphene is a two-dimensional one-atom-thick planar sheet of sp(2)-bonded carbon atoms while CNTs are a cylindrical nanostructure which composed entirely of sp(2)-bonded carbon atoms as well. This review presents and discusses the past and current advancement of synthesis and characterization of graphene and CNTs. The review also concludes with a brief summary and an outlook on the challenges and future prospects in the growth of graphene and CNTs.
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The effects of multiwalled carbon nanotubes on epitheliocytes of different compartments of the gastrointestinal tract and urothelium of different compartments of the renal nephron were studied in CBA mice. The nanotubes affected mouse gastrointestinal mucosa and renal urothelium. The cell reaction in the macula densa of the renal distal tubules and the immune system reaction to oral nanotubes were detected. A possible effect of nanotubes administered orally on the renal filtration function was hypothesized.
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Use of rare earth elements (REEs) has increased rapidly in recent decades due to technological advances. It has been accompanied by recurring rare earth element anomalies in water bodies. In this work we (i) studied the effects of eight novel doped and one non-doped rare earth oxide (REO) particles (aimed to be used in solid oxide fuel cells and gas separation membranes) on algae, (ii) quantified the individual adverse effects of the elements that constitute the (doped) REO particles and (iii) attempted to find a discernible pattern to relate REO particle physicochemical characteristics to algal growth inhibitory properties. Green algae Raphidocelis subcapitata (formerly Pseudokirchneriella subcapitata) were used as a test species in two different formats: a standard OECD201 algal growth inhibition assay and the algal viability assay (a 'spot test') that avoids nutrient removal effects. In the 24h 'spot' test that demonstrated direct toxicity, algae were not viable at REE concentrations above 1mgmetal/L. 72-hour algal growth inhibition EC50 values for four REE salts (Ce, Gd, La, Pr) were between 1.2 and 1.4mg/L, whereas the EC50 for REO particles ranged from 1 to 98mg/L. The growth inhibition of REEs was presumably the result of nutrient sequestration from the algal growth medium. The adverse effects of REO particles were at least in part due to the entrapment of algae within particle agglomerates. Adverse effects due to the dissolution of constituent elements from (doped) REO particles and the size or specific surface area of particles were excluded, except for La2NiO4. However, the structure of the particles and/or the varying effects of oxide composition might have played a role in the observed effects. As the production rates of these REO particles are negligible compared to other forms of REEs, there is presumably no acute risk for aquatic unicellular algae.
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We systematically investigated the toxicity mechanism of three graphene-family materials (GFMs), graphene oxide (GO), reduced graphene oxide (rGO) and multi-layer graphene (MG), to algae (Chlorella pyrenoidosa). GFMs exhibited much higher toxicity than other carbon materials (carbon nanotube and graphite), with the 96 h median effective concentration (EC50) values of 37.3 (GO), 34.0 (rGO), and 62.2 (MG) mg/L. Shading effect contributed approximately 16.4% of growth inhibition by GO due to its higher dispersibility and transformation while the other GFMs did not show any shading effect. Hydrophobic rGO and MG more readily heteroagglomerated with algae than GO, thus likely leading to more direct contacts with algae. Flow cytometry results showed significant decrease of membrane integrity after GFM exposure, and rGO caused the highest membrane damage, which was confirmed by the increased DNA and K⁺ efflux. The observed membrane damage was caused by a combination of oxidative stress and physical penetration/extraction. Moreover, all the three GFMs could adsorb macronutrients (N, P, Mg, and Ca) from the algal medium, thus leading to nutrient depletion-induced indirect toxicity. GO showed the highest nutrient depletion (53% of total toxicity) due to its abundant functional groups. The information provided in this work will be useful for understanding toxicity mechanism and environmental risk of different GFMs in aquatic environments.
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Nano-particles (NPs) are used in industrial and biomedical fields such as cosmetics, food additives and biosensors. Beside their favorable properties, nanoparticles are responsible for toxic effects. Local adverse effects and/or systemic toxicity are described with nanoparticle delivery to target organs of the human body. Animal studies provide evidence for the aforementioned toxicity. Cardiac function is a specific target of nanoparticles. Thus, reviewing the current bibliography on cardiotoxicity of nanoparticles and specifically of titanium, zinc, silver, carbon, silica and iron oxide nano-materials is the aim of this study.
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The use of metal based nanoparticles (NPs) is increasing which leads to their release in water bodies via various waste streams and warrants risk assessment. Consistent biological effect data on NPs for environmentally relevant test species that are accompanied by thorough characterization of NPs are scarce but indispensable for understanding possible risks of NPs. We composed and tested a library of 12 metal-based nanoparticles (Al2O3, Co3O4, CuO, Fe3O4, MgO, Mn3O4, Sb2O3, SiO2, ZnO, TiO2, WO3 and Pd) using alga Pseudokirchneriella subcapitata, three bacterial species (Vibrio fischeri, Escherichia coli, Staphylococcus aureus) and protozoan Tetrahymena thermophila. The NPs were characterized for physico-chemical properties, solubility and abiotic reactive oxygen species (ROS) production. Also, respective soluble salts were analysed for toxic effects. The algal growth inhibition assay proved the most sensitive and yielded EC50 values for 10 NPs ranging from 0.1 to 58 mg/l. Algal toxicity correlated with abiotic ROS production of NPs and the majority of NPs formed agglomerates that entrapped algal cells. Despite of different sensitivity, there was a common trend in the toxicity of NPs across different species and test formats: CuO and ZnO had highest toxicity (EC50 values below 1 mg/l) to all organism groups except protozoa. The high toxicity was mostly due to shedding of toxic concentrations of Zn and Cu ions; for most of the test species Al2O3, SiO2, WO3 and Sb2O3 were not toxic below 100 mg/l and MgO showed no adverse effect below 100 mg/l to any test species in any test setting.
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Ubiquitous dissolved organic matter (DOM) from various sources can interact with discharged multiwalled carbon nanotubes (MWCNTs) and pose distinct effects on the toxicity of MWCNTs to aquatic organisms. This study for the first time compared effects of a natural originated humic acid (HA) and two synthetic surfactants (sodium dodecylbenzenesulfonate – SDBS, octyl phenoxy polyethoxyethanol – TX100) on the toxicity of MWCNTs to a unicellular green alga (Chlorella pyrenoidosa). Algal growth inhibition, cell surface hydrophobicity, oxidative stress, MWCNT-cell agglomeration, and cell morphology change and internalization of MWCNTs were assayed to investigate individual and combined toxicities of MWCNTs and DOMs, and mechanisms underlying different effects of DOMs on the toxicity of MWCNTs were specifically addressed. It is shown that SDBS and TX100 were capable of promoting cell internalization of MWCNTs and triggering higher oxidative stress, and thereby increasing the toxicity of MWCNTs; while HA could alleviate the toxicity of MWCNTs through limiting cell internalization of MWCNTs and reducing oxidative stress. The outcomes of this work shed new light on the assessment of ecological toxicity of discharged nanomaterials.
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The biological response to pristine and annealed multi-walled carbon nanotubes (MWCNT) was assessed on murine macrophages (RAW 264.7). First, the physicochemical features of the as-produced MWCNT and annealed at 2125 °C for 1 h were fully characterized. A decrease in structural defects, hydrophobicity and catalytic impurities was detected after annealing. Thereafter, their impact on cytotoxicity, oxidative stress, and pro-inflammatory response was investigated at concentrations ranging from 15 to 120 µg mL−1. No effect of the 2125 °C treatment was detected on the cytotoxicity. In contrast, the annealed carbon nanotubes showed a significant increase of the pro-inflammatory response. We assumed that this behavior was due to the reduction in structural defects that may modify the layer of adsorbed biomolecules. Surprisingly, the purification of metallic catalysts did not have any significant impact on the oxidative stress. We suggested that the structural improvements from the 2125 °C treatment can decrease the carbon nanotube scavenging capacity and thus allow a higher free radical release which may counterbalance the decrease of oxidative stress due to a lower content of metallic impurities. Graphical Abstract
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The increasing industrial use of nanomaterials during the last decades poses a potential threat to the environment and in particular to organisms living in the aquatic environment. In the present study, the toxicity of zinc oxide nanoparticles (ZnO NPs) was investigated in Marine algae Chlorella vulgaris (C. vulgaris). High zinc dissociation from ZnONPs, releasing ionic zinc in seawater, is a potential route for zinc assimilation and ZnONPs toxicity. To examine the mechanism of toxicity, C. vulgaris were treated with 50mg/L, 100mg/L, 200mg/L and 300mg/L ZnO NPs for 24h and 72h. The detailed cytotoxicity assay showed a substantial reduction in the viability dependent on dose and exposure. Further, flow cytometry revealed the significant reduction in C. vulgaris viable cells to higher ZnO NPs. Significant reductions in LDH level were noted for ZnO NPs at 300mg/L concentration. The activity of antioxidant enzyme superoxide dismutase (SOD) significantly increased in the C. vulgaris exposed to 200mg/L and 300mg/L ZnO NPs. The content of non-enzymatic antioxidant glutathione (GSH) significantly decreased in the groups with a ZnO NPs concentration of higher than 100mg/L. The level of lipid peroxidation (LPO) was found to increase as the ZnO NPs dose increased. The FT-IR analyses suggested surface chemical interaction between nanoparticles and algal cells. The substantial morphological changes and cell wall damage were confirmed through microscopic analyses (FESEM and CM). Copyright © 2014 Elsevier Inc. All rights reserved.
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Here multi-walled carbon nanotubes (MWNT) of different average diameters with ppm level purity were produced by a low temperature C2H4 pyrolysis on Fe-Co type catalysts combined with forthcoming graphitization at 2200–2800 °C in argon flow. Annealed nanotubes were characterized with X-ray fluorescent analysis, BET surface measurements, HR TEM, X-ray diffraction, DTA analysis, and measurements of temperature and magnetic field dependences of conductivity. The graphitization of MWNT results in removal of residual catalyst metal impurities, reduction of the wall defects, and closure of nanotube tips. It was found that extent of these effects depends on MWNT diameters. It was proposed that the graphitization is caused by a significant thermal displacement and diffusion of carbon atoms at temperatures higher the Debye temperature.
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Carbon nanotubes (CNT) are strong sorbents for organic micropollutants, but changing environmental conditions may alter the distribution and bioavailability of the sorbed substances. Therefore, we investigated the effect of green algae (Chlorella vulgaris) on sorption of a model pollutant (diuron, synonyms: 3-(3,4-Dichlorophenyl)-1,1-dimethylurea, DCMU) to CNT (multi-walled purified, industrial grade, pristine, and oxidized; reference material: Diesel soot). In absence of algae, diuron sorption to CNT was fast, strong, and nonlinear (Freundlich coefficients: 10(5.79)-10(6.24) μg/kgCNT·(μg/L)(-n) and 0.62-0.70 for KF and n, respectively). Adding algae to equilibrated diuron-CNT mixtures led to 15-20% (median) diuron re-dissolution. The relatively high amorphous carbon content slowed down ad-/desorption to/from the high energy sorption sites for both industrial grade CNT and soot. The results suggest that diuron binds readily, but - particularly in presence of algae - partially reversibly to CNT, which is of relevance for environmental exposure and risk assessment.
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Some researchers consider nanotechnology the next industrial revolution, and consumer products and a variety of industries increasingly use synthetic nanoparticles. In this Account, we review the initial accomplishments of nanoecotoxicology, a discipline that is just a decade old. This new subdiscipline of ecotoxicology faces two important and challenging problems: the analysis of the safety of nanotechnologies in the natural environment and the promotion of sustainable development while mitigating the potential pitfalls of innovative nanotechnologies. In this Account, we provide a snapshot of the publicly available scientific information regarding the ecotoxicity of engineered nanoparticles. We pay special attention to information relevant to aquatic freshwater species commonly used for risk assessment and regulation.
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Increasing studies focus on nanotoxicity, but many unknowns are remaining to be investigated. This study examined toxicities of four varieties of oxide nanoparticles (Al2O3, SiO2, ZnO, and TiO2) to the green algae Chlorella sp. Nanoparticulate Al2O3, SiO2, and TiO2 (DJ3, rutile) had no significant toxicity, whereas nano-ZnO and nano-TiO2 (HR3, anatase) greatly inhibited the algal growth with 6 d EC30 of ca. 20 and 30 mg L−1, respectively. Comparative experiments showed the algal toxicities decreased with an order of Zn2+ > nano-ZnO > bulk-ZnO when the concentration of the test material was lower than 50 mg L−1. However, nano-ZnO had higher algal toxicity than Zn2+ ions at concentrations >50 mg L−1 and released less Zn2+ ions into the culture media than bulk-ZnO, which suggests that dissolved Zn2+ ions from nano-ZnO were not the dominant mechanism for the algal growth inhibition. Shading effects of nano-ZnO and HR3 were also excluded from the main mechanism of the nanotoxicity by the comparison of the nanotoxicities in the presence and absence of illumination. Large aggregates of nano-ZnO and HR3 entrapping and wrapping the algal cells were observed, which may contribute to the nanotoxicity. These results will be helpful for understanding the toxicity of oxide nanoparticles.
Article
The ichthyotoxic red tide organism Heterosigma akashiwo (Raphidophyceae) has been associated with fish kill events within the aquaculture industry for many years. The precise toxicological mechanism involved in these fish kills is unclear; however, much research attention has focused on the production of reactive oxygen species (ROS) by these toxic algae. In this study, we investigated the production of hydrogen peroxide (H2O2) by isolates of H. akashiwo and the nontoxic chlorophyte Tetraselmis apiculata. Subsequently, we tested those concentrations of H2O2 on vertebrate cell lines and the invertebrate Artemia salina (brine shrimp) to investigate mortality. Net production rates for the H. akashiwo isolates ranged from 0.46 to 7.89 pmol H2O2 min-1 (104 cells)-1 while obtaining maximum concentrations between 0.14 and 0.91 μM H2O2. Conversely, T. apiculata produced only 0.03 pmol H2O2 min-1 (104 cells)-1 with a maximum level on 0.04 μM. However, toxic effects on UMR-106 and HEK-293 cells were only induced by acute and protracted exposure to concentrations of H2O2 ≥ 0.1 mM. Additionally, significant mortality of A. salina in the presence or absence of ferric and ferrous iron was induced by H2O2 levels ≥ 1 mM. Iron is a redox metal that reduces H2O2 to hydroxy radicals. These data collectively indicate that production of H2O2 by multiple isolates of H. akashiwo is orders of magnitude less than that required for mortality of either the vertebrate cell lines or the invertebrate A. salina. Other nonichthyotoxic roles for extracellular ROS are proposed.
Article
Aim: Double-walled carbon nanotubes (DWNTs) are found in a variety of consumer products, but there are no ecotoxicity data of DWNTs into marine organisms. Materials & methods: Chronic toxicity of DWNTs was investigated with the diatom Thalassiosira pseudonana, copepod Tigriopus japonicus and medaka Oryzias melastigma. DWNTs were dispersed using sonication (so-DWNTs) and stirring (st-DWNTs) for comparison. Results: The median aggregation size (0.89 m2) of so-DWNTs was smaller than that of st-DWNTs (21.8 m 2). Exposure to DWNTs led to growth inhibition of T. pseudonana with EC50s of 1.86 and 22.7 mg/l for so- and st-DWNTs, respectively. Population growth of T. japonicus was reduced to 0.1 mg/l for so-DWNTs and 10 mg/l for st-DWNTs. Growth inhibition in O. melastigma was observed at 10 mg/l for so-DWNTs but not for st-DWNTs. Conclusion: Given that so-DWNTs are consistently significantly more toxic than st-DWNTs, dispersion method and size of aggregations should be considered in DWNT toxicity testing. © 2010 Future Medicine Ltd.
Article
It has become clear over the last 15–20 years that the immediate effect of a wide range of environmental stresses, and of infection, on vascular plants is to increase the formation of reactive oxygen species (ROS) and to impose oxidative stress on the cells. Since 1994, sufficient examples of similar responses in a broad range of marine macroalgae have been described to show that reactive oxygen metabolism also underlies the mechanisms by which seaweeds respond (and become resistant) to stress and infection. Desiccation, freezing, low temperatures, high light, ultraviolet radiation, and heavy metals all tend to result in a gradual and continued buildup of ROS because photosynthesis is inhibited and excess energy results in the formation of singlet oxygen. The response to other stresses (infection or oligosaccharides which signal that infection is occurring, mechanical stress, hyperosmotic shock) is quite different—a more rapid and intense, but short‐lived production of ROS, described as an “oxidative burst”—which is attributed to activation of NADPH oxidases in the plasma membrane. Seaweed species that are able to survive such stresses or resist infection have the capacity to remove the ROS through a high cellular content of antioxidant compounds, or a high activity of antioxidant enzymes.
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Due to growing production, carbon nanotubes (CNT) may soon be found in a broad range of products and thus in the environment. In this work, an algal growth test was developed to determine effects of pristine and oxidized CNT on the green algae Chlorella vulgaris and Pseudokirchneriella subcapitata. CNT suspensions were prepared in algal test medium and characterized taking into account the suspension age, the reduced light transmittance of nanoparticle suspensions defined as shading of CNT and quantified by UV/vis spectroscopy, and the agglomeration of the CNT and of the algal cells. Growth inhibition and photosynthetic activity were investigated as end points. Growth of C. vulgaris was inhibited with effect concentrations of 50% (EC(50)) values of 1.8 mg CNT/L and of 24 mg CNT/L in well dispersed and in agglomerated suspensions, respectively, and 20 mg CNT/L and 36 mg CNT/L for P. subcapitata, respectively. However, the photosynthetic activity was not affected. Growth inhibition was highly correlated with the shading of CNT and the agglomeration of algal cells. This suggests that the reduced algal growth might be caused mainly by indirect effects, i.e. by reduced availability of light and different growth conditions caused by the locally elevated algal concentration inside of CNT agglomerates.
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The multiwalled carbon nanotubes (MWNTs) are novel materials with many potential applications. The ecotoxicity of these materials is not well studied, but it is essential for environmental impact assessments. In this study a commercially available MWNT material was carboxylated by microwave assisted acid oxidation. This functionalized MWNT (f-MWNT) material was examined for toxicity effects using unicellular marine green alga Dunaliella tertiolecta. D. tertiolecta was exposed to f-MWNT which had been pre-equilibrated with culture media for 24 h. Substantial growth lag phase was observed at 5 and 10 mgL(-1) f-MWNT, and the resulting 50% effective concentration (EC50) on 96-h growth was 0.82 ± 0.08 mgL(-1). During mid-exponential growth phase cytotoxicity was evidenced at 10 mgL(-1) f-MWNT in 36% reduction in exponential growth rate, 88 mV more positive glutathione redox potential (indicative of oxidative stress), 5% and 22% reduction in photosystem II (PSII) quantum yield and functional cross section respectively, all relative to the control cultures. However, when the large f-MWNT aggregates in the media with 10 mgL(-1) f-MWNT were removed by 0.2 μm filtration, D. tertiolecta did not show significant cytotoxicity effects in any of the above parameters. This suggests that the cytotoxicity effects originated predominantly from the large f-MWNT aggregates. Analysis of the f-MWNT aggregation dynamics suggests active interaction between f-MWNT and algal cells or cell metabolites that promoted f-MWNT aggregation formation. The f-MWNT particles were also found absorbed on algal cell surface. The direct contact between f-MWNT and cell surface was likely responsible for reduced PSII functional cross section and oxidative stress during exponential growth.
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Toxicities of ZnO, TiO2 and CuO nanoparticles to Pseudokirchneriella subcapitata were determined using OECD 201 algal growth inhibition test taking in account potential shading of light. The results showed that the shading effect by nanoparticles was negligible. ZnO nanoparticles were most toxic followed by nano CuO and nano TiO2. The toxicities of bulk and nano ZnO particles were both similar to that of ZnSO4 (72 h EC50 approximately 0.04 mg Zn/l). Thus, in this low concentration range the toxicity was attributed solely to solubilized Zn2+ ions. Bulk TiO2 (EC50=35.9 mg Ti/l) and bulk CuO (EC50=11.55 mg Cu/l) were less toxic than their nano formulations (EC50=5.83 mg Ti/l and 0.71 mg Cu/l). NOEC (no-observed-effect-concentrations) that may be used for risk assessment purposes for bulk and nano ZnO did not differ (approximately 0.02 mg Zn/l). NOEC for nano CuO was 0.42 mg Cu/l and for bulk CuO 8.03 mg Cu/l. For nano TiO2 the NOEC was 0.98 mg Ti/l and for bulk TiO2 10.1 mg Ti/l. Nano TiO2 formed characteristic aggregates entrapping algal cells that may contribute to the toxic effect of nano TiO2 to algae. At 72 h EC50 values of nano CuO and CuO, 25% of copper from nano CuO was bioavailable and only 0.18% of copper from bulk CuO. Thus, according to recombinant bacterial and yeast Cu-sensors, copper from nano CuO was 141-fold more bioavailable than from bulk CuO. Also, toxic effects of Cu oxides to algae were due to bioavailable copper ions. To our knowledge, this is one of the first systematic studies on effects of metal oxide nanoparticles on algal growth and the first describing toxic effects of nano CuO towards algae.