The removal of antibiotics has attracted much attention due to their extremely high adverse impacts on the environment. However, the potential risks of degradation intermediates are seldom reported. In this work, the influence of different factors on the electro-catalytic degradation efficiency of tetracycline hydrochloride (TCH) by the prepared carbon nanotubes/agarose/indium tin oxide (CNTs/AG/ITO) electrode was investigated. Under optimal conditions (10 wt% CNTs dosage, pH = 7), the maximum degradation efficiency for TCH (10 mg L⁻¹) reached up to 96% within 30 min treatment with 4 V potential. Superoxide anions (•O₂⁻) played an important role in the electro-catalytic degradation. Totally 10 degradation intermediates were identified using HPLC-MS/MS, and the degradation pathway was proposed. Toxicities of the parent antibiotic and the identified intermediates were calculated using the ECOSAR (Ecological Structure Activity Relationship) program in EPISuite, and results showed that more toxic intermediates were generated. The maximal chronic toxicity for green algae of the intermediate increased 1439.92 times. Furthermore, antimicrobial activity was further verified by disk agar biocidal tests with Escherichia coli ATCC25922 and higher biotoxicity intermediates compared with parent compounds were confirmed to be formed. Therefore, more attention should be paid on the potential risk of degradation intermediates in the treatment of wastewater containing antibiotics.

Antibiotic resistance in environmental matrices becomes urgently significant for public health and has been considered as an emerging environmental contaminant. In this work, the ampicillin-resistant Escherichia coli (AR E. coli) and corresponding resistance genes (blaTEM₋₁) were effectively eliminated by the electrocatalytic process, and the dissemination risk of antibiotic resistance was also investigated. All the AR E. coli (∼8 log) was inactivated and 8.17 log blaTEM₋₁ was degraded by the carbon nanotubes/agarose/titanium (CNTs/AG/Ti) electrode within 30 min. AR E. coli was inactivated mainly attributing to the damage of cell membrane, which was attacked by reactive oxygen species and subsequent leakage of intracellular cytoplasm. The blaTEM₋₁ was degraded owing to the strand breaking in the process of electrocatalytic degradation. Furthermore, the dissemination risk of antibiotic resistance was effectively controlled after being electrocatalytic treatment. This study provided an effective electrocatalytic technology for the inactivation of antibiotic resistant bacteria and control of antibiotic resistance dissemination risk in the aqueous environment.

The development of nanotechnology has increased the amount of nanoparticles in the environment inducing pollution. In view of increasing amounts, their toxicity assessment becomes important. Aluminum oxide nanoparticles (Al₂O₃ NPs) have a wide range of applications in industry. The present study aims to reveal the time-dependent (24, 48, 72, 96 h) and dose-dependent (0, 5, 25, 50 mg/ml) effects of 13-nm-sized Al₂O₃ NPs on an agronomic plant wheat (Triticum aestivum L.) roots correlating with the appearance of various cellular stress responses. Al₂O₃ NPs reduced the root elongation by 40.2 % in 5 mg/ml, 50.6 % in 25 mg/ml, and 54.5 % in 50 mg/ml after 96 h. Histochemical analysis revealed lignin accumulation, callose deposition, and cellular damage in root cortex cells correlating the root elongation inhibition. Although the nanoparticle application decreased the total protein content with respect to control after 96 h, the peroxidase activity increased significantly which is considered to be one of the oxidative stress factors. Moreover, agarose gel results revealed that Al₂O₃ NPs induced DNA fragmentation being one of the important markers of programmed cell death. In conclusion, direct exposure to Al₂O₃ NPs leads to phytotoxicity significantly in wheat roots culminating in morphological, cellular, and molecular alterations.

The diffusive gradient in thin film technique was recently adapted to organic compounds. The diffusional coefficient (D) is a key parameter needed to calculate the time-weighted average concentration. In this study, two methods are used for D measurement in two gels (agarose and polyacrylamide): the diffusion cell method (Dcₑₗₗ) and the slice stacking method (Dₛₜₐcₖ). Thus, D were discussed and compared for 112 organic compounds, including pesticides, hormones, and pharmaceuticals. Dₛₜₐcₖ tends to be higher than Dcₑₗₗ. It could be explained by the presence of a non-negligible diffusive boundary layer thickness in diffusion cell. Consequently, the use of sampling rates (RS) should be more adequate to determine water concentration, for a given bulk flow velocity. Dₛₜₐcₖ also corresponds to the diffusion in gel only, allowing the determination of the maximal RS, and would be considered as a reference value that can be adjusted to in situ conditions, by applying the appropriate DBL thickness. The range and variability of D values found in the literature and obtained in this work were discussed. Relationships between D and compound physicochemical properties (molecular mass, log Dow, polar surface area, van der Waals volume) were investigated. We did not find clear and robust correlation between D and any single physicochemical property, for the set of compounds tested.

The residual antibiotics in different environmental media pose a serious threat to human health and the ecosystem. The high-efficient elimination of antibiotics is one of the foremost works. In this study, chloramphenicol (CAP) was eliminated efficiently by electrocatalytic advanced oxidation process with carbon nanotubes/agarose/indium tin oxide (CNTs/AG/ITO) electrode. The influences of different experimental parameters on the degradation efficiency were systematically studied. Under the optimal conditions (4 V potential, 10 wt% CNTs dosage, and pH = 10), the maximum degradation efficiency of CAP (20 mg L⁻¹) achieved 88% within 180 min. Besides, the electrocatalytic degradation pathway and mechanism for CAP were also investigated, •O₂⁻ played a major role in the process of electrocatalytic degradation. Based on the QSAR (quantitative structure-activity relationship) model, the toxicities of CAP and identified intermediates were analyzed. Compared with the parent compound, the maximal chronic toxicity of intermediate ((E)-3-(4-nitrophenyl)prop-1-ene-1,3-diol) for daphnid increased 197-fold. Besides, the hybrid toxicity of the degradation system was further confirmed via disk agar biocidal tests with Escherichia coli ATCC25922, which changed slightly during the degradation process. Based on the above results, it is worth noting that the degradation pathway and toxicity assessment should be paid more attention to the treatment of antibiotic wastewater.

Heat- and pH-stable phytase efficiently hydrolyzes phytic acid. In this research, heat- and pH-stable mutant phytases, T⁸³R, L²⁸⁷R, and T⁸³R/L²⁸⁷R were generated by site-directed mutagenesis from Yersinia intermedia. After the induction and expression of recombinant wild-type and mutant phytases in E. coli BL21, the enzymes were purified using nickel sepharose affinity chromatography, and characterized kinetically and thermodynamically using spectroscopy methods. The mutants showed optimum activity at pH 5.15 and 55–61 °C. The catalytic efficiencies of T⁸³R, L²⁸⁷R, T⁸³R/L²⁸⁷R, and wild-type phytases were calculated to be 2941, 29346, 4906, and 6917 mmol/L⁻¹s⁻¹, respectively. Moreover, after the incubation of T⁸³R, L²⁸⁷R, wild-type, and T⁸³R/ L²⁸⁷R phytases at 100 °C for 1 h, the enzymes retained 22, 5, 4, and 2% of their initial activities, respectively. In addition, T⁸³R, T⁸³R/L²⁸⁷R, L²⁸⁷R, and wild-type phytases retained 82, 44, 16 as well as 11% of their initial activities after 1 h at pH 5.15, respectively. Among these mutants, T⁸³R mutant showed 18% increase in thermal stability, 71% increase in pH stability, and +0.103 KJ/mole increase in ΔΔG, while the catalytic efficiency and ΔΔG value of L²⁸⁷R mutant increased by 4 times and +0.0903 KJ/mole, respectively. Thus, the mutants have the potential to be used in feed industries to increase the bioavailability of minerals while decreasing soil and water pollution.

The gill tissue is the main site of metabolic enzymes or compensation, with the kidney tissue playing a supporting role. At the gill tissue, carbonic anhydrase enzymes (CAs) catalyze the hydration of CO₂ to HCO₃⁻ and H⁺ for production to the H₂O. In this work, the CA enzyme was purified from horse mackerel (Trachurus trachurus) gill with a specific activity of 21,381.42 EU/mg, purification fold of 150.61, total activity of 2347.68 EU/mL, and a yield of 16.13% using sepharose 4B-L-tyrosine-sulfanilamide affinity gel chromatography. For recording the enzyme purity, gel electrophoresis was performed, and single band was seen. The molecular weight of this enzyme was found approximately 35 kDa. Also, the inhibitory effects of different pesticides such as thiram, clofentezine, propineb, deltamethrin, azoxystrobin, and thiophanate and heavy metal ions such as Fe²⁺, Cu²⁺, Co²⁺, Pb²⁺ Hg²⁺, and As³⁺ on horse mackerel gill tissue CA enzyme activities were investigated. Our results indicated that these pesticides and metal ions showed inhibitory effects at low nanomolar and millimolar concentrations for fish gill CA enzymes, respectively.

Metal oxide nano-particles have definitely unique toxicological properties than currently investigated oxides. Therefore, this study was aimed to comparatively evaluate the genotoxicity of nano-CuO and bulk CuO particles on a model fish species Oreochromis niloticus. Fish were exposed to two selected concentrations (¹/₁₀ and ¹/₂₀ of the LC50/96 h) of both nano-CuO and bulk CuO for 30 days. Genotoxic effects associated with DNA ladder formation and chromosomal damage were investigated using DNA fragmentation and micronucleus techniques. Based on DNA fragmentation of fish hepatocytes, the two selected concentrations of bulk and nano-CuO were found to induce DNA damage. Analysis of the DNA fragments initiated by bulk CuO on agarose gel revealed DNA ladder pattern, which is commonly considered as a hallmark of apoptosis, while fish exposed to nano-CuO particles showed a molecular hallmark of necrosis which is the fragmentation of the nuclear DNA into a smear-like pattern. Also, DNA damage was further confirmed quantitatively using the image analysis software Image J. In this context, nano-CuO-treated groups exhibited a maximum DNA damage especially at the lower concentration (¹/₂₀ LC50/96 h). To ensure CuO genotoxicity, micronucleus and other nine nuclear abnormalities were studied in peripheral erythrocytes and significant (p < 0.05) elevation was observed in nano-CuO-exposed groups at the lower concentration followed by a decrease in extent of chromosomal damage at the higher concentration, while fish groups treated with bulk CuO showed a more or less dose-dependent effect.

Passive samplers based on diffusive gradients in thin hydrogel films (DGT) were recently modified for sampling of polar organic compounds in water. However, since the sampling rates of the commonly used DGT design with the surface area of 3.1 cm² are low, we propose to increase them by applying a two-sided design with a larger sampling surface area of 22.7 cm². The sampler design consists of two sorptive hydrogel disks compressed between two diffusive hydrogel disk layers strengthened by nylon netting and held together by two stainless steel rings. Sorbent/water distribution coefficients (KSW) were determined, and the sampler was calibrated for monitoring 11 perfluoroalkyl substances and 12 pharmaceuticals and personal care products in water at laboratory conditions using a closed system with artificial flow generated by submersible pumps. A field performance test was conducted at five locations in the Morava River basin in Czech Republic. The median value of laboratory-derived sampling rates was 43 mL day⁻¹ with extreme values of 2 mL day⁻¹ and 90 mL day⁻¹ for perfluorotridecanoic and perfluoroheptanoic acids, respectively. The log KSW values of tested compounds ranged from 3.18 to 5.47 L kg⁻¹, and the estimated halftime to attain sampler-water equilibrium ranged from 2 days to more than 28 days, which is the maximum recommended exposure period, considering potential issues with the stability of hydrogel. The sampler can be used for assessment of spatial trends as well as estimation of aqueous concentration of investigated polar compounds.

This study deals with further and systematic laboratory evaluation of the already known ammonium 12-molybdophosphate (AMP)-diffusive gradient in thin film (DGT) method, which is used for measuring total Cs concentration in environmental waters. This study confirms that the AMP-binding gel is not stable for pH > 6. In order to reveal a potential impact of AMP degradation on DGT application, time-series experiments were performed by deploying AMP-DGT samplers in Cs-doped moderately basic soft and hard water up to total AMP-binding gel degradation (60 and 175 h of deployment time, respectively). Linear accumulation of Cs by AMP-DGT samplers was observed up to 48 and 58 h in hard and soft waters, respectively. For this deployment time range, AMP-DGT measured over 77 ± 10 and 94 ± 16% of total Cs concentration in hard and soft water, respectively. The difference in DGT response was attributed to Ca²⁺ and Mg²⁺ competition reducing the uptake of AMP-DGT samplers in hard water. Shrinkage of agarose-polyacrylamide diffusive gel was experimentally observed only in hard water due to more intensive AMP-binding gel degradation in hard water. Even if the AMP-DGT response was not impacted in this study, it is recommended to use agarose hydrogel as standard diffusive gel. Based on the experience obtained from this detailed validation process, the authors propose a number of key requirements that need to be considered when developing DGT devices, with testing the performance over longer deployment times being critical. Graphical abstract ᅟ