Despite the increasing use of rare earth elements (REEs) and oxides (REOs) in various technologies, the information on their ecotoxicological hazard is scarce. Here, the effects of La³⁺, Ce³⁺, Pr³⁺, Nd³⁺, Gd³⁺, CeO₂, and eight doped REOs to marine bacteria Vibrio fischeri and freshwater protozoa Tetrahymena thermophila were studied in parallel with REO dopant metals (Co²⁺, Fe³⁺, Mn²⁺, Ni²⁺, Sr²⁺). The highest concentrations of REOs tested were 100 mg/L with protozoa in deionized water and 500 mg/L with bacteria in 2% NaCl. Although (i) most REOs produced reactive oxygen species; (ii) all studied soluble REEs were toxic to bacteria (half-effective concentration, EC₅₀ 3.5–21 mg metal/L; minimal bactericidal concentration, MBC 6.3–63 mg/L) and to protozoa (EC₅₀ 28–42 mg/L); and (iii) also some dopant metals (Ni²⁺, Fe³⁺) proved toxic (EC₅₀ ≤ 3 mg/L), no toxicity of REOs to protozoa (EC₅₀ > 100 mg/L) and bacteria (EC₅₀ > 500 mg/L; MBC > 500 mg/L) was observed except for La₂NiO₄ (MBC 25 mg/L). According to kinetics of V. fischeri bioluminescence, the toxicity of REEs was triggered by disturbing cellular membrane integrity. Fortunately, as REEs and REOs are currently produced in moderate amounts and form in the environment insoluble salts and/or oxides, they apparently present no harm to aquatic bacteria and protozoa.

The effects of microplastics (MP) are extensively studied, yet hazard data from long-term exposure studies are scarce. Moreover, for sustainable circular use in the future, knowledge on the biological impact of recycled plastics is essential. The aim of this study was to provide long-term toxicity data of virgin vs recycled (mechanical recycling) low density polyethylene (LDPE) for two commonly used ecotoxicity models, the freshwater crustacean Daphnia magna and the terrestrial crustacean Porcellio scaber. LDPE MP was tested as fragments of 39.8 ± 8.82 µm (virgin) and 205 ± 144 µm (recycled) at chronic exposure levels of 1–100 mg LDPE/L (D. magna) and 0.2–15 g LDPE/kg soil (P. scaber). Mortality, reproduction, body length, total lipid content, feeding and immune response were evaluated. With the exception of very low inconsistent offspring mortality at 10 mg/L and 100 mg/L of recycled LDPE, no MP exposure-related adverse effects were recorded for D. magna. For P. scaber, increased feeding on non-contaminated leaves was observed for virgin LDPE at 5 g/kg and 15 g/kg. In addition, both LDPE induced a slight immune response at 5 g/kg and 15 g/kg with more parameters altered for virgin LDPE. Our results indicated different sublethal responses upon exposure to recycled compared to virgin LDPE MP.

Secretory phospholipasesA₂ (sPLA₂s) form a large family of structurally related enzymes widespread in nature. Herein, we studied the inhibitory effects of sPLA₂s from Vipera lebetina (VLPLA₂), Vipera berus berus (VBBPLA₂), and Naja naja oxiana (NNOPLA₂) venoms on (i) human platelets, (ii) four different bacterial strains (gram-negative Escherichia coli and Vibrio fischeri; gram-positive Staphylococcus aureus and Bacillus subtilis) and (iii) five types of cancer cells (PC-3, LNCaP, MCF-7, K-562 and B16-F10) in vitro. sPLA₂s inhibited collagen-induced platelet aggregation: VBBPLA₂ IC₅₀ = 0.054, VLPLA₂ IC₅₀ = 0.072, NNOPLA₂ IC₅₀ = 0.814 μM. p-Bromophenacylbromide-inhibited sPLA₂ had no inhibitory action on platelets. 36.17 μM VBBPLA₂ completely inhibited the growth of gram-positive Bacillus subtilis whereas no growth inhibition was observed towards gram-negative Escherichia coli. The inhibitory action of sPLA₂s (~0.7 μM and ~7 μM) towards cancer cells depended on both venom and cell type. VBBPLA₂ (7.2 μM) inhibited significantly the viability of K-562 cells and the cell death appeared apoptotic. The sPLA₂s exhibited no inhibitory effect towards LNCaP cells and some effect (8%–20%) towards other cells. Thus, already sub-μM concentrations of sPLA₂s inhibited collagen-induced platelet aggregation and from the current suite of studied svPLA₂s and test cells, VBBPLA₂ was the most growth inhibitory towards Bacillus subtilis and K-562 cells.

Clinical use of CuO nanoparticles (NPs) as antibacterials can be hampered by their toxicity to human cells. We hypothesized that certain surface functionalizations of CuO NPs may render NPs toxic to bacteria, but still be relatively harmless to human cells. To control this hypothesis, the toxicity of differently functionalized CuO NPs to bacteria Escherichia coli vs human cells (THP-1 macrophages and HACAT keratinocytes) was compared using similar conditions and end points. CuO NPs functionalized with polyethylene glycol (CuO–PEG), carboxyl (CuO–COOH, anionic), ammonium (CuO–NH₄⁺, cationic) and unfunctionalized CuO NPs and CuSO₄ (controls) were tested. In general, the toxicity of Cu compounds decreased in the following order: CuO–NH₄⁺ > unfunctionalized CuO > CuSO₄ > CuO–COOH > CuO–PEG. Positively charged unfunctionalized CuO and especially CuO–NH₄⁺ proved most toxic (24-h EC₅₀ = 21.7–47 mg/l) and had comparable toxicity to bacterial and mammalian cells. The multivariate analysis revealed that toxicity of these NPs was mostly attributed to their positive zeta potential, small hydrodynamic size, high Cu dissolution, and induction of reactive oxygen species (ROS) and TNF-α. In contrast, CuO–COOH and CuO–PEG NPs had lower toxicity to human cells compared to bacteria despite efficient uptake of these NPs by human cells. In addition, these NPs did not induce TNF-α and ROS. Thus, by varying the NP functionalization and Cu form (soluble salt vs NPs), it was possible to “target” the toxicity of Cu compounds, whereas carboxylation and PEGylation rendered CuO NPs that were more toxic to bacteria than to human cells envisaging their use in medical antibacterial products.

Bacterial levansucrases polymerize fructose residues of sucrose to β-2,6 linked fructans—fructooligosaccharides (FOS) and levan. While β-2,1-linked FOS are widely recognized as prebiotics, the health-related effects of β-2,6 linked FOS are scarcely studied as they are not commercially available. Levansucrase Lsc3 (Lsc-3) of Pseudomonas syringae pv. tomato has very high catalytic activity and stability making it a promising biotechnological catalyst for FOS and levan synthesis. In this study we evaluate feasibility of several high-throughput methods for screening and preliminary characterization of levansucrases using 36 Lsc3 mutants as a test panel. Heterologously expressed and purified His-tagged levansucrase variants were studied for: (1) sucrose-splitting activity; (2) FOS production; (3) ability and kinetics of levan synthesis; (4) thermostability in a Thermofluor assay. Importantly, we show that sucrose-splitting activity as well as the ability to produce FOS can both be evaluated using permeabilized levansucrase-expressing E. coli transformants as catalysts. For the first time we demonstrate the key importance of Trp109, His113, Glu146 and Glu236 for the catalysis of Lsc3. Cost-effective and high-throughput methods presented here are applicable not only in the levansucrase assay, but have a potential to be adapted for high-throughput (automated) study of other enzymes.