Toxicity of nanoparticulate and bulk ZnO, Al2O3 and TiO2 to the nematode Caenorhabditis elegans

Limited information is available on the environmental behavior and associated potential risk of manufactured oxide nanoparticles (NPs). In this research, toxicity of nanoparticulate and bulk ZnO, Al₂O₃ and TiO₂ were examined to the nematode Caenorhabditis elegans with Escherichia coli as a food source. Parallel experiments with dissolved metal ions from NPs were also conducted. The 24-h median lethal concentration (LC₅₀) and sublethal endpoints were assessed. Both NPs and their bulk counterparts were toxic, inhibiting growth and especially the reproductive capability of the nematode. The 24-h LC₅₀ for ZnO NPs (2.3 mg L⁻¹) and bulk ZnO was not significantly different, but significantly different between Al₂O₃ NPs (82 mg L⁻¹) and bulk Al₂O₃ (153 mg L⁻¹), and between TiO₂ NPs (80 mg L⁻¹) and bulk TiO₂ (136 mg L⁻¹). Oxide solubility influenced the toxicity of ZnO and Al₂O₃ NPs, but nanoparticle-dependent toxicity was indeed observed for the investigated NPs.     

Assessment of cytotoxicity and toxicity for phosphonium-based deep eutectic solvents

In this work, the cytotoxicity and toxicity of phosphonium-based deep eutectic solvents (DESs) with three hydrogen bond donors, namely glycerine, ethylene glycol, and triethylene glycol were investigated. The cytotoxicity effect was tested using brine shrimp (Artemia salina). The toxicity was investigated using the two Gram positive bacteria Bacillus subtilis and Staphylococcus aureus, and two Gram negative bacteria Escherichia coli and Pseudomonas aeruginosa. The cytotoxicity of tested DESs was much higher than that of their individual components, indicating their toxicological behavior was different. It was also found that there was toxic effect on the studied bacteria, indicating their potential application as anti-bacterial agents. To the best of our knowledge, this is the first time the cytotoxicity and toxicity of phosphonium-based DESs were studied.     

Antimicrobial activity of metal oxide nanoparticles supported onto natural clinoptilolite

The antimicrobial activity of Cu₂O, ZnO and NiO nanoparticles supported onto natural clinoptilolite was investigated in the secondary effluent under dark conditions. After 24 h of contact the Cu₂O and ZnO nanoparticles reduced the numbers of viable bacterial cells of Escherichia coli and Staphylococcus aureus in pure culture for four to six orders of magnitude and showed consistent 100% of antibacterial activity against native E. coli after 1 h of contact during 48 exposures. The antibacterial activity of NiO nanoparticles was less efficient. The Cu₂O and NiO nanoparticles showed 100% of antiprotozoan activity against Paramecium caudatum and Euplotes affinis after 1 h of contact, while ZnO nanoparticles were less efficient. The morphology and crystallinity of the nanoparticles were not affected by microorganisms. The metal oxide nanoparticles could find a novel application in the disinfection of secondary effluent and removal of pathogenic microorganisms in the tertiary stage of wastewater treatment.     

Effect of chlorination and ultraviolet disinfection on tetA-mediated tetracycline resistance of Escherichia coli

Antibiotic-resistant bacteria are an emerging threat to public health during drinking water consumption and reclaimed water reuse. Several studies have shown that the proportions of antibiotic-resistant bacteria in waters may increase when exposed to low doses of UV light or chlorine. In this study, inactivation of tetracycline-resistant Escherichia coli and antibiotic-sensitive E. coli by UV disinfection and chlorination was compared to determine the tolerance of tetracycline-resistant E. coli to UV light and chlorine, and tetracycline resistance of a tetracycline-resistant E. coli population was studied under different doses of the disinfectants. Our results showed that relative to antibiotic-sensitive E. coli, tetracycline-resistant E. coli had the same tolerance to UV light and a potentially higher tolerance to chlorination. The mortality frequency distributions of tetracycline-resistant E. coli exposed to tetracycline were shifted by both chlorination and UV disinfection. When compared to the hemi-inhibitory concentrations (IC₅₀) of tetracycline-resistant E. coli with no exposure to UV or chlorination, the IC₅₀ of tetracycline-resistant E. coli treated with tetracycline was 40% lower when inactivation by UV light or chlorination reached 3-log but was 1.18 times greater when inactivation by chlorination reached 4.3-log. Chlorination applied to drinking water or reclaimed water treatment may increase the risk of selection for highly tetracycline-resistant E. coli.     

Ecotoxicity of nanoparticles of CuO and ZnO in natural water

The acute toxicity of CuO and ZnO nanoparticles in artificial freshwater (AFW) and in natural waters to crustaceans Daphnia magna and Thamnocephalus platyurus and protozoan Tetrahymena thermophila was compared. The L(E)C₅₀ values of nanoCuO for both crustaceans in natural water ranged from 90 to 224 mg Cu/l and were about 10-fold lower than L(E)C₅₀ values of bulk CuO. In all test media, the L(E)C₅₀ values for both bulk and nanoZnO (1.1-16 mg Zn/l) were considerably lower than those of nanoCuO. The natural waters remarkably (up to 140-fold) decreased the toxicity of nanoCuO (but not that of nanoZnO) to crustaceans depending mainly on the concentration of dissolved organic carbon (DOC). The toxicity of both nanoCuO and nanoZnO was mostly due to the solubilised ions as determined by specific metal-sensing bacteria.     

Toxicity of imidazolium- and pyridinium-based ionic liquids and the co-metabolic degradation of N-ethylpyridinium tetrafluoroborate

We examined the effects of the ionic liquids (ILs), 1-butyl-3-methylimidazolium hexafluorophosphate [BMIM][PF₆], N-ethylpyridinium tetrafluoroborate [EtPy][BF₄], and N-ethylpyridinium trifluoroacetate [EtPy][CF₃COO] on Pseudomonas fluorescens, a ubiquitous soil bacterium. In the presence of 0.5- and 1% of [BMIM][PF₆] or [EtPy][CF₃COO] the growth of bacteria was inhibited, whereas exposing them to 1% [EtPy][BF₄] increased the lag period wherein bacteria adapt to growth conditions before continuing to grow. However, at higher concentrations (5% and 10%), no growth was observed. The inhibitory effects were evident by a decrease in the optical density of the culture, a decline in the consumption of the carbon source, citric acid, and a change in the size of the bacterium. At concentrations below 1%, [EtPy][BF₄] was metabolized by P. fluorescens in the presence of citric acid. Oxidation of the side alkyl-chain of [EtPy][BF₄] caused the accumulation of N-hydroxylethylpyridinium and pyridinium as major degradation products.     

Are deep eutectic solvents benign or toxic?

In continuation of investigation for environmentally benign protocol for new solvents termed deep eutectic solvents (DESs), it is herein reported results concerning the toxicity and cytotoxicity of choline chloride (ChCl) based DESs with four hydrogen bond donors including glycerine, ethylene glycol, triethylene glycol and urea. The toxicity was investigated using two Gram positive bacteria Bacillus subtilis and Staphylococcus aureus, and two Gram negative bacteria Escherichia coli and Pseudomonas aeruginosa. The cytotoxicity effect was tested using the Artemia salina leach. It was found that there was no toxic effect for the tested DESs on all of the studied bacteria confirming their benign effects on these bacteria. Nevertheless, it was found that the cytotoxicity of DESs was much higher than their individual components (e.g. glycerine, ChCl) indicating that their toxicological behavior is different. For our best knowledge this is the first time that toxicity and cytotoxicity of DESs were studied. The toxicity and cytotoxicity of DESs varied depending on the structure of components. Careful usage of the terms non-toxicity and biodegradability must be considered. More investigation on this matter is required.     

Ecotoxicity and biodegradability of antielectrostatic dicephalic cationic surfactants

Four series of dicephalic cationic surfactants, considered as new antielectrostatic agents have been investigated in order to establish their toxicity and biodegradability. Among them N,N-bis[3,3′-(dimethylamine)propyl]alkylamides, N,N-bis[3,3′-(dimethylamine)propyl]alkylamide dihydrochlorides, N,N-bis[3,3′-(trimethylammonio)propyl]alkylamide dibromides and N,N-bis[3,3′-(trimethylammonio)propyl]alkylamide dimethylsulphates with different hydrophobic chain length (n-C₉H₁₉ to n-C₁₅H₃₁) and type of counterion (chloride, bromide and methylsulfate) have been studied. The inhibitory effect against microorganisms has been examined using model gram-positive and gram-negative bacteria, and yeasts. None of the tested surfactants have shown antimicrobial activity against gram-negative bacteria (Escherichia coli, Pseudomonas putida) and yeasts (Saccharomyces cerevisiae, Rhodotorula glutinis) at a concentration below 1000 μg mL⁻¹, however some of them were moderately active against gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis). The Microtox® test was successfully applied to measure EC₅₀ values of the studied dicephalic cationic surfactants. Their toxicity to Vibrio fischeri depended upon the alkanoyl chain length with the EC₅₀ values in a range of 2.6-980 mg L⁻¹. N,N-bis[3,3′-(dimethylamine)propyl]alkylamide dihydrochlorides 2a-b and N,N-bis[3,3′-(trimethylammonio)propyl]alkylamide dibromides 3a-b comprising n-decanoyl and n-dodecanoyl hydrophobic tails appeared to be the least toxic. Furthermore, the biodegradability under aerobic conditions of 2a-b, 3a-b was evaluated using OECD Method 301F. According to the obtained results 2a, 3a-3b can be considered as almost readily biodegradable and they are not expected to be persistent in the environment. Additionally, partial biodegradation was observed for 2b, indicating its possible biodegradation in wastewater treatment systems.     

Controllable synthesis Fe3O4@POHABA core-shell nanostructure as high-performance recyclable bifunctional magnetic antimicrobial agent

We demonstrated a method to form magnetic antimicrobial POHABA (poly-N,N′-[(4,5-dihydroxy-1,2-phenylene)bis(methylene)]bisacrylamide)-based core-shell nanostructure by free-radical polymerization of OHABA on the Fe₃O₄ core surface. The magnetic antimicrobial agent Fe₃O₄@POHABA can be used in domestic water treatment against bacterial pathogens. The thickness of POHABA shell could be controlled from 10.4 ± 1.2 to 56.3 ± 11.7 nm by the dosage of OHABA. The results of antimicrobial-activity test indicated that POHABA-based core-shell nanostructure had broad-spectrum inhibitory against Gram-negative, Gram-positive bacteria and fungi. The minimum inhibitory concentration (MIC) values of Fe₃O₄@POHABA nanostructure against Escherichia coli and Bacillus subtilis were both 0.4 mg/mL. Fe₃O₄@POHABA nanostructures responded to a permanent magnet and were easily recycled. Fe₃O₄@POHABA nanoparticles retained 100% antimicrobial efficiency for both Gram-negative and Gram-positive bacteria throughout eight recycle procedures.

     

In vivo genotoxicity assessment of titanium,zirconium and aluminium nanoparticles,and their microparticulated forms,in Drosophila

As in vivo system, we propose Drosophila melanogaster as a useful model for study the genotoxic risks associated with nanoparticle exposure. In this study we have carried out a genotoxic evaluation of titanium dioxide (TiO₂), zirconium oxide (ZrO₂) and aluminium oxide (Al₂O₃) nanoparticles and their microparticulated forms in D. melanogaster by using the wing somatic mutation and recombination assay. This assay is based on the principle that loss of heterozygosis and the corresponding expression of the suitable recessive markers, multiple wing hairs and flare-3, can lead to the formation of mutant clones in treated larvae, which are expressed as mutant spots on the wings of adult flies. Third instar larvae were feed with TiO₂, ZrO₂ and Al₂O₃ nanoparticles, and their microparticulated forms, at concentrations ranging from 0.1 to 10 mM. Although a certain level of aggregation/agglomeration was observed in solution, it must be noted than the constant digging activity of larvae ensures that treated medium pass constantly through the digestive tract ensuring exposure. The results showed that no significant increases in the frequency of all spots (e.g. small single, large single, twin, total mwh and total spots) were observed, indicating that these nanoparticles were not able to induce genotoxic activity in the wing spot assay of D. melanogaster. Negative data were also obtained with the microparticulated forms. This indicates that the nanoparticulated form of the selected nanomaterials does not modify the potential genotoxicity of their microparticulated versions. These in vivo results contribute to increase the genotoxicity database on the TiO₂, ZrO₂ and Al₂O₃ nanoparticles.     

Visible light photocatalytic antibacterial activity of Ni-doped and N-doped TiO<Subscript>2</Subscript> on <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and <Emphasis Type="Italic">Escherichia coli</Emphasis> bacteria

The Ni-doped and N-doped TiO₂ nanoparticles were investigated for their antibacterial activities on Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) bacteria. Their morphological features and characteristics such as particle size, surface area, and visible light absorbing capacity were compared and discussed. Scanning electron microscopy, X-ray diffraction, and UV–visible spectrophotometry were used to characterize both materials. The inactivation of E. coli (as an example of Gram-negative bacteria) and S. aureus (as an example of Gram-positive bacteria) with Ni-doped and N-doped TiO₂ was investigated in the absence and presence of visible light. Antibacterial activity tests were conducted using undoped, Ni-doped, and N-doped TiO₂. The N-doped TiO₂ nanoparticles show higher antibacterial activity than Ni-doped TiO₂. The band gap narrowing of N-doped TiO₂ can induce more visible light absorption and leads to the superb antibacterial properties of this material. The complete inactivation time for E. coli at an initial cell concentration of 2.7?×?10⁴ CFU/mL was 420 min which is longer than the 360 min required for S. aureus inactivation. The rate of inactivation of S. aureus using the doped TiO₂ nanoparticles in the presence of visible light is greater than that of E. coli. The median lethal dose (LD50) values of S. aureus and E. coli by antibacterial activity under an 18-W visible light intensity were 80 and 350 mg/ml for N-doped TiO₂, respectively.     

Effect of ingested titanium dioxide nanoparticles on the digestive gland cell membrane of terrestrial isopods

The aim of this study was to find out whether ingested titanium dioxide nanoparticles (nano-TiO₂) cause cell membrane damage by direct contact or by lipid peroxidation. We assessed lipid peroxidation and digestive gland cell membrane stability of animals fed on food dosed with nano-TiO₂. Conventional toxicity measures were completed to determine if cellular effects are propagated to higher levels of biological complexity. An invertebrate model organism (Porcellio scaber, Isopoda, Crustacea) was fed with food containing nanosized TiO₂ and the result confirmed that at higher exposure concentrations after 3 d exposure, nano-TiO₂ destabilized cell membranes but lipid peroxidation was not detected. Oxidative stress as evidenced by lipid peroxidation was observed at longer exposure durations and high exposure doses. These data suggest that cell membranes are destabilized by direct interactions between nanoparticles and cell membrane, not solely via oxidative stress.     

Unexpected toxic interactions in the freshwater amphipod Gammarus pulex (L.) exposed to binary copper and nickel mixtures

To document the toxicity of copper and nickel in binary mixtures, freshwater amphipods Gammarus pulex were exposed to the metals given independently or as mixtures. Toxicity to Cu alone was relatively high: 96-h LC₁₀ and LC₅₀ were found at 91 and 196 μg L^?1, respectively. Toxicity to Ni alone was very low, with 96-h LC₁₀ and LC₅₀ of 44,900 and 79,200 μg L^?1, respectively. Mixture toxicities were calculated from single toxicity data using conventional models. Modeled toxicity was then compared with the measured toxicity of the binary mixture. Two kinds of mixtures were tested. Type I mixtures were designed as combinations of Cu and Ni given at the same effect concentrations, when taken independently, to identify possible interactions between copper and nickel. In type II mixtures, Cu concentrations varied from 0 to 600 μg L^?1 while the nickel concentration was kept constant at 500 μg L^?1 to mimic conditions of industrial wastewater discharges. Ni and Cu showed synergic effects in type I mixtures while type II mixtures revealed antagonistic effects. Low doses of Ni reduced Cu toxicity towards G. pulex. These results show that even for simple binary mixtures of contaminants with known chemistry and toxicity, unexpected interactions between the contaminants may occur. This reduces the reliability of conventional additivity models.     

Distinctive effects of nano-sized permethrin in the environment

Pesticides are an essential tool in integrated pest management. Nanopermethrin was prepared by solvent evaporation from an oil-in-water volatile microemulsion. The efficacy of the formulated nanopermethrin was tested against Aedes aegypti and the results compared to those of regular, microparticular permethrin. The 24 h LC₅₀ for nanopermethrin and permethrin was found to be 0.0063 and 0.0199 mg/L, respectively. The formulated nanopermethrin was tested for toxicity against non-target organisms. Nanopermethrin did not show antibacterial activity against Escherichia coli (ATCC 13534 and 25922) or against Bacillus subtilis. Phytotoxicity studies of nanopermethrin to the seeds of Lycopersicum esculentum, Cucumis sativus, and Zea mays showed no restraint in root length and germination percentage. In the Allium cepa test, regular microparticular permethrin treatment of 0.13 mg/L showed a mitotic index (MI) of 46.8 % and chromosomal aberration of 0.6 %, which was statistically significant (p?<?0.05) compared to control. No significant differences were observed in 0.13 mg/L nanopermethrin exposure as compared to control (MI of 52.0 and 55.03 % and chromosomal aberration of 0.2 and 0 %, respectively). It was concluded that formulated nanopermethrin can be used as a safe and effectual alternative to commercially available permethrin formulation in agricultural practices.     

Phytotoxicity of nanoparticles—problems with bioassay choosing and sample preparation

For a full estimation of the risk related with the presence of engineered nanoparticles (ENPs) in the environment, the use of the current ecotoxicological methods may prove insufficient. In the study presented herein, various methods of assessment of ecotoxicity were applied to compare the phytotoxicity of three ENPs: nano-ZnO, nano-TiO₂ and nano-Ni. The toxicity was assayed both for aqueous solutions of the ENPs (the germination/elongation test and Phytotestkit F^TM) and for ENPs added to soil (Phytotoxkit F^TM and modified Phytotoxkit F^TM). Lepidium sativum was used as a test plant. The scope of the study also included the assessment of the effect of the method of ENP application to the soil (as powder and aqueous suspension) on their phytotoxicity. In the course of the study, no effect of the studied ENPs and their bulk counterparts on the germination of seeds was observed. The root growth inhibition of L. sativum depended on the kind of test applied. The trend between concentration of ENPs and effect depended on the method used and kind of ENPs. For most nanoparticles (despite of the method used), the differences in phytotoxicity between nano and bulk particles were observed. Depending on the kind of ENPs, their phytotoxicity differs between water and soil. ZnO (nano and bulk) and nano-Ni were more toxic in soil than in water. For TiO₂ and bulk-Ni, reverse trend was observed. A different method of ENP application to soil differently affects the phytotoxicity.     

The effect of γ-Fe2O3 nanoparticles on Escherichia coli genome

Extensive production and application of γ-Fe₂O₃ magnetic nanoparticles (MNPs) has increased their potential risk on environment and human health. This report illustrates a genetic impact of γ-Fe₂O₃ magnetic nanoparticles (MNPs) on Escherichia coli (E. coli). After 3000-generation incubation with MNPs addition, obvious genomic variations were revealed by using repetitive extragenic palindromic PCR (rep-PCR) DNA fingerprint technique. The physicochemical interactions between MNPs and bacteria could be responsible for such genomic responses. It was revealed that Fe³⁺ concentration increased in the medium. Transmission electronic microscopy (TEM) and flow cytometry (FCM) analysis consistently demonstrated the occurrences of adsorption and membranes-internalization of MNPs outside and inside cells. Both increased Fe³⁺ ion and the uptake of MNPs facilitated Fe binding with proteins and DNA strands, resulting in enhancing the mutation frequency of E. coli. Our results would be of great help to assessing the potential impact of MNPs on human and environment.     

Novel application of the CORAL software to model cytotoxicity of metal oxide nanoparticles to bacteria Escherichia coli

Convenient to apply and available on the Internet software CORAL (http://www.insilico.eu/CORAL) has been used to build up quantitative structure-activity relationships (QSAR) for prediction of cytotoxicity of metal oxide nanoparticles to bacteria Escherichia coli (minus logarithm of concentration for 50% effect pEC50). In this study six random splits of the data into the training and test set were examined. It has been shown that the CORAL provides a reliable tool that could be used to build up a QSAR of the pEC50.     

Semi-specific Microbacterium phyllosphaerae-based microbial sensor for biochemical oxygen demand measurements in dairy wastewater

Although the long incubation time of biochemical oxygen demand (BOD₇) measurements has been addressed by the use of microbial biosensors, the resulting sensor-BOD values gained from the measurements with specific industrial wastewaters still underestimates the BOD value of such samples. This research aims to provide fast and more accurate BOD measurements in the dairy wastewater samples. Unlike municipal wastewater, wastewater from the dairy industry contains many substrates that are not easily accessible to a majority of microorganisms. Therefore, a bacterial culture, Microbacterium phyllosphaerae, isolated from dairy wastewater was used to construct a semi-specific microbial biosensor. A universal microbial biosensor based on Pseudomonas fluorescens, which has a wide substrate spectrum but is nonspecific to dairy wastewater, was used as a comparison. BOD biosensors were calibrated with OECD synthetic wastewater, and experiments with different synthetic and actual wastewater samples were carried out. Results show that the semi-specific M. phyllosphaerae-based microbial biosensor is more sensitive towards wastewaters that contain milk derivates and butter whey than the P. fluorescens-based biosensor. Although the M. phyllosphaerae biosensor underestimates the BOD₇ value of actual dairy wastewaters by 25–32 %, this bacterial culture is more suitable for BOD monitoring in dairy wastewater than P. fluorescens, which underestimated the same samples by 46–61 %.     

Toxicity of TiO2, ZrO2, Fe,Fe2O3, and Mn2O3 nanoparticles to the yeast, Saccharomyces cerevisiae

The growing application of engineered nanomaterials is leading to an increased occurrence of nanoparticles (NPs) in the environment. Thus, there is a need to better understand their potential impact on the environment. This study evaluated the toxicity of nanosized TiO₂, ZrO₂, Fe⁰, Fe₂O₃, and Mn₂O₃ towards the yeast Saccharomyces cerevisiae based on O₂ consumption and cell membrane integrity. In addition, the state of dispersion of the nanoparticles in the bioassay medium was characterized.     

Rapid biological synthesis of silver nanoparticles using <Emphasis Type="Italic">Leucas martinicensis</Emphasis> leaf extract for catalytic and antibacterial activity

A novel green approach for the synthesis and stabilization of silver nanoparticles (AgNPs) using water extract of Leucas martinicensis leaf has been developed. As obtained, the nanoparticles are characterized by UV-visible (UV-Vis), transmission electron microscope (TEM), and X-ray diffraction (XRD). The crystalline nature of the AgNPs is confirmed by the prominent peaks in the XRD pattern. FTIR spectra suggest that the possible biomolecules are responsible for the efficient stabilization of the sample. The effects of leaf quantity on the biosynthesis of AgNPs are investigated by UV-Vis spectrophotometer. The synthesized AgNPs are observed to have a good catalytic activity on the reduction of methylene blue by L. martinicensis leaf. This is confirmed by the decrease in absorbance maximum values of methylene blue with respect to time through UV-Vis spectrophotometer. Moreover, the antibacterial activity of synthesized AgNPs against Staphylococcus aureus, Bacillus subtilis, Salmonella typhi, and Escherichia coli are screened.