Compound-specific isotope analysis (CSIA) is an established tool to track the in situ transformation of organic chemicals at contaminated sites. In this work, we evaluated the potential of multi-element CSIA to assess biodegradation of 2,3-dichloroaniline (2,3-DCA), which is a major industrial feedstock. Using controlled laboratory experiments, we determined, for the first time, negligible carbon (<0.5‰) and hydrogen (<10‰) isotope fractionation and a significant inverse nitrogen isotope fractionation (>10‰) during aerobic 2,3-DCA biodegradation by a mixed enrichment culture. The tentative identification of a glutamate conjugate of 2,3-DCA as a reaction intermediate indicates that the initial multistep enzymatic reaction may be rate-limiting. The formation of the glutamate adduct would increase the bond energy at the N atom, thus likely explaining the observed inverse N isotope fractionation. The corresponding nitrogen enrichment factor was +6.8 ± 0.6‰. This value was applied to investigate the in situ 2,3-DCA biodegradation at a contaminated site where the carbon and nitrogen isotope signatures from field samples suggested similar aerobic processes by native microorganisms. Under the assumption of the applicability of the Rayleigh model in a pilot wetland treating contaminated groundwater, the extent of biodegradation was estimated to be up to 80-90%. This study proposes multi-element CSIA as a novel application to study 2,3-DCA fate in groundwater and surface water and provides insights into biodegradation pathways.

Aniline and its derivate are critical environmental pollutants, and thus, the introduction of an eco-friendly catalyst for removing them is an important research future. The ZnO supported on the ball-mill prepared clinoptilolite nanoparticles (CNPs) was prepared via an ion-exchange process followed by the calcination process. The amount of loaded ZnO in the ZnO-CNP (CZ) samples varied as 0.54, 0.63, 0.72, and 0.86 meq/g as the Zn(II) concentration in the ion-exchange solution varied from 0.1 to 0.5 M. The ZnO-CNP catalyst was briefly characterized by XRD, FTIR, and DRS techniques. The pHpzc value for the various ZnO-CNPs was about 7.1 that had no change with the ZnO loading. By applying the Scherrer equation on the XRD results, a nano-dimension of about 50 nm was obtained for the catalyst. Bandgap energy of the ZnO-CNP samples was estimated by applying the Kubelka-Munk equation on the DRS reflectance spectra. The value for the CZ2 catalyst was about 3.64 eV. The supported ZnO-CNP sample was then used in the photodegradation of 2,4-dichloroaniline (DCA). Raw zeolite showed a relatively low photocatalytic activity. The degradation efficiency was followed by recording the absorbance of the DCA solution by UV-Vis spectrophotometer. The effects of the essential critical operating factors on the degradation efficiency were kinetically studied by applying the Hinshelwood equation to the results. The ZnO-CNP catalyst with 2 w% ZnO showed the best photocatalytic rate in the optimal conditions of 0.75 g/L, CDCA: 15 ppm, and the initial pH: 5.8. Finally, HPLC analysis of the blank and the photodegraded DCA solutions at 180 and 300 min confirmed 74 and 87% of DCA molecules were degraded during these times. The results confirm that supported ZnO onto clinoptilolite caused enhanced photocatalytic activity because the zeolite internal electrical field prevents the e-/h+ recombination.

The ball-mill clinoptilolite nanoparticles (CNP) was ion-exchanged in Ni(II) solutions and calcined to obtain NiO-CNP catalysts with various NiO loadings. The resultant CNP was ion-exchanged in 0.1, 0.2, 0.3, and 0.4 M Ni(II) solutions and then calcined at 450 °C. The resultant NiO-CNPs contained 1.9, 2.3, 3.0, and 3.2% NiO, respectively. The XRD, FTIR, and DRS characterization techniques were applied. By applying the Scherrer equation on the XRD results, the average crystallite size for the NiO-CNP samples was estimated in the range of 42-65 nm. The pHpzc of the NiO-CNP species was slightly changed from 6.8 to 7.6 by an increase in the loaded NiO. The band gap energy of the samples was calculated by applying the Kubelka-Munk equation on the DRS results. The band gap energies of 3.81, 4.05, and 3.63 eV were estimated for the direct electronic transitions of the CN2, CN2.3, and CN3.2 samples, respectively. The boosted photoactivity was obtained in 2,4-dichloroanilyne (DCA) degradation when NiO supported onto both micronized clinoptilolite and its nanoparticles. The effects of the most important experimental variables on DCA photodegradation rate were kinetically studied by applying the Hinshelwood model on the results. The faster rate for the DCA photodegradation was achieved at the optimal conditions, including the catalyst dose: 0.5 g/L, CDCA: 5 ppm, and the initial pH: 3. Some new peaks were observed in the HPLC chromatograms for the photodegraded DCA solutions after 180 min and 300 min, which showed 84% and 95% DCA photodegradation.

Fish embryos are excellent models for studies aimed at understanding toxic mechanisms and indications of possible acute and chronic effects. For the past 3 yr, an Arabian killifish (Aphanius dispar) fish embryo test has been developed in the authors\' laboratory as a routine ecotoxicological test that can be used to support risk assessment of potential contaminants in Arabian Gulf coastal waters. Tests were conducted with 3 reference toxicants (3,4-dichloroaniline [DCA], sodium dodecyl sulfate, and zinc sulfate [Zn]) and chlorine, a disinfectant used widely in industrial cooling systems around the Arabian Gulf region. The 50% effect concentration (EC50) for DCA was 0.47 mg/L and 1.89 mg/L for embryos exposed before 6 hpf and after 168 hpf, respectively. Sublethal effects were mainly observed at concentrations above 2.5 mg/L, the effects included severe pericardial edema and tail shortage. The sodium dodecyl sulfate ionic surfactant caused mortality at both early and late stages of embryo development; it caused coagulation, severe deformity, and hemolysis. Both the EC50 and the 50% lethal concentration (LC50) for sodium dodecyl sulfate were 9.37 mg/L. Salinity influenced the toxicity of Zn to killifish embryos: at 40 psu Zn was found not to be toxic, whereas at 20 psu toxicity had increased significantly (p < 0.05). Values of EC50 and LC50 were 2.5 mg/L and 4 mg/L, respectively. Concentrations above 15 mg/L in embryos were often accompanied by upper abdominal edema and inhibition of growth, especially evident in the tail. Chlorine caused mortality at a lower concentration; for example, at 0.05 mg/L 33% of embryos were found dead at the end of the experiment. The LC50 for chlorine was determined to be 0.08 mg/L. Examination of the existing literature showed similar results to the present study\'s findings. The results suggest a more comparable sensitivity of killifish embryos to that of other fish embryo test recommended species. The present study\'s findings support the ability of killifish to be an indicator organism for environmental risk assessments of Arabian Gulf waters. Benefits include sensitivity to a wide range of substances and conditions, animal alternative, ease of fish breeding, and clarity of the embryos.

Soil organic amendment addition is an effective practice in Mediterranean areas due to its associated high agricultural benefits and its potential to reduce the pesticide impact on water resources. However, their metabolites have received scarce attention, even when they may pose more risk than their parent compounds. Two winery vermicomposts obtained from spent grape marc (V1) and the mixture vine shoot-biosolid vinasses (V2) have been investigated as low cost organic amendments to minimize the leaching of diuron, imidacloprid and their metabolites in columns packed with a sandy loam (S1) and a silty-clay loam soil (S2) under steady state flow conditions. In the unamended soil columns, leached amounts of diuron were 75% and 53% in S1 and S2, respectively. Its metabolites (3-(3,4-dichlorophenyl)-1-methylurea, DPMU; and 3,4-dichlorophenylurea, DPU) percolated less than 35% of the total applied amount. The amount of the metabolite 3,4-dichloroaniline (DCA) was 2% and 30% for S1 and S2, respectively. Leaching of imidacloprid was 79% and 96% for S1 and S2, respectively, while its metabolite 6-chloronicotinic acid (CNA) was entirely leached. In the vermicompost-amended columns, the leaching of diuron was reduced 2 to 3-fold. DPMU and DPU were also significantly reduced (more than 6-fold). DCA did not appear in any of the leachates of the amended soil columns. Imidacloprid leaching was reduced 1 to 2-folds in the amended columns. The amendments did not affect the transport of CNA. The dissolved organic carbon (DOC) from the vermicomposts did not enhance pesticide transport throughout the soil in any case. This qualitative study presents these vermicomposts as an effective potential low-cost tool in reducing pesticide and metabolite leaching. The next step would be to test them under more realistic conditions.