Toxicity impact of hydrogen peroxide on the fate of zebrafish and antibiotic resistant bacteria.

Hydrogen peroxide (H2O2) is applied in various environments. It could be present at concentrations ranging from nanomolar to micromolar in a water system. It is produced through pollutants and natural activities. Since few studies have been conducted about the impact of naturally produced H2O2 on aquatic organisms, the objective of the present study was to monitor changes in responses of aquatic model organisms such as zebrafish and antibiotic-resistant bacteria to different exogenous H2O2 exposure. Increases in exposure concentration and time induced decreases in the perception of zebrafish larvae (up to 69%) and movement of adult zebrafish (average speed, average acceleration, movement distance, and activity time) compared to the control (non-exposed group). In addition, as a function of H2O2 exposure concentration (0-100,000 nM) and time, up to 20-fold increase (p = 5.00*10-6) of lipid peroxidation compared to control was observed. For microorganisms, biofilm, an indirect indicator of resistance to external stressors, was increased up to 68% and gene transfer was increased (p = 2.00*10-6) by more than 30% after H2O2 exposure. These results imply that naturally generated H2O2 could adversely affect aquatic environment organisms and public health. Thus, more careful attention is needed for H2O2 production in an aquatic system.

Metabolite tracking to elucidate the effects of environmental pollutants.

The purpose of this study was to determine whether behavioral tests and metabolic profiling of organisms can be promising alternatives for assessing the health of aquatic systems. Water samples from four potential pollution sources in South Korea were collected for toxicity evaluation. First, conventional acute toxicity test in Daphnia magna and behavioral test in zebrafish was conducted to assess water quality. Second, metabolomic analysis was performed on zebrafish exposed to water samples and on environmental fish collected from the same source. Acute toxicity test in D. magna showed that none of the water samples exerted significant adverse effects. However, activity of zebrafish larvae exposed to samples from the zinc smelter (ZS) and industrial complex (IND) sites decreased compared to those exposed to samples from the reference site (RS). Metabolomic analysis using the Manhattan plot and Partial Least Square (PLS)/Orthogonal PLS Discriminant Analysis (OPLS-DA) showed differences in metabolic profiles between RS and ZS, and between IND and abandoned mine site (M). Interestingly, applying the same metabolomic analysis to environmental fish revealed patterns similar to those for zebrafish, despite the uncontrollable variables involved in environmental sampling. This study shows that metabolomics is a promising tool in assessing the health of aquatic environments.

Sub-lethal pharmaceutical hazard tracking in adult zebrafish using untargeted LC-MS environmental metabolomics.

Antibiotics in the aquatic environment are dispersed through anthropogenic activities at low concentrations. Despite their sub lethal concentration, these biologically active compounds may still have adverse effects to non-target species. This study examined the response of adult zebrafish to 0.1mg/L concentration of clarithromycin, florfenicol, sulfamethazine, and their mixture using environmental metabolomics. Embryo and larvae of the fish were also used to assess fish embryo acute toxicity and behavior tests respectively. The fish embryo toxicity test did not show any inhibition of growth and development of the embryos after 96h of exposure to the antibiotics. Changes in swimming activity were seen in 5-dpf larvae which is believed to be correlated with the length of exposure to the compounds. Meanwhile, environmental metabolomics revealed diverse metabolites and pathways that were affected after 72h of exposure of the adult fish to sub-lethal concentration of the compounds. We found that even at low concentration of the antibiotics, behavioral and metabolic effects were still observed despite the lack of visible morphological changes. Further studies involving other aquatic organisms and bioactive compounds are encouraged to strengthen the findings presented in this novel research.

Significance of metabolite extraction method for evaluating sulfamethazine toxicity in adult zebrafish using metabolomics.

Recently, environmental metabolomics has been introduced as a next generation environmental toxicity method which helps in evaluating toxicity of bioactive compounds to non-target organisms. In general, efficient metabolite extraction from target cells is one of the keys to success to better understand the effects of toxic substances to organisms. In this regard, the aim of this study is (1) to compare two sample extraction methods in terms of abundance and quality of metabolites and (2) investigate how this could lead to difference in data interpretation using pathway analysis. For this purpose, the antibiotic sulfamethazine and zebrafish (Danio rerio) were selected as model toxic substance and target organism, respectively. The zebrafish was exposed to four different sulfamethazine concentrations (0, 10, 30, and 50mg/L) for 72h. Metabolites were extracted using two different methods (Bligh and Dyer and solid-phase extraction). A total of 13,538 and 12,469 features were detected using quadrupole time-of-flight liquid chromatography mass spectrometry (QTOF LC-MS). Of these metabolites, 4278 (Bligh and Dyer) and 332 (solid phase extraction) were found to be significant after false discovery rate adjustment at a significance threshold of 0.01. Metlin and KEGG pathway analysis showed comprehensive information from fish samples extracted using Bligh and Dyer compared to solid phase extraction. This study shows that proper selection of sample extraction method is critically important for interpreting and analyzing the toxicity data of organisms when metabolomics is applied.

Evaluation of acidity estimation methods for mine drainage, Pennsylvania, USA.

Eighteen sites impacted by abandoned mine drainage (AMD) in Pennsylvania were sampled and measured for pH, acidity, alkalinity, metal ions, and sulfate. This study compared the accuracy of four acidity calculation methods with measured hot peroxide acidity and identified the most accurate calculation method for each site as a function of pH and sulfate concentration. Method E1 was the sum of proton and acidity based on total metal concentrations; method E2 added alkalinity; method E3 also accounted for aluminum speciation and temperature effects; and method E4 accounted for sulfate speciation. To evaluate errors between measured and predicted acidity, the Nash-Sutcliffe efficiency (NSE), the coefficient of determination (R (2)), and the root mean square error to standard deviation ratio (RSR) methods were applied. The error evaluation results show that E1, E2, E3, and E4 sites were most accurate at 0, 9, 4, and 5 of the sites, respectively. Sites where E2 was most accurate had pH greater than 4.0 and less than 400 mg/L of sulfate. Sites where E3 was most accurate had pH greater than 4.0 and sulfate greater than 400 mg/L with two exceptions. Sites where E4 was most accurate had pH less than 4.0 and more than 400 mg/L sulfate with one exception. The results indicate that acidity in AMD-affected streams can be accurately predicted by using pH, alkalinity, sulfate, Fe(II), Mn(II), and Al(III) concentrations in one or more of the identified equations, and that the appropriate equation for prediction can be selected based on pH and sulfate concentration.

Transfer of antibiotic resistance plasmids in pure and activated sludge cultures in the presence of environmentally representative micro-contaminant concentrations.

The presence of antibiotics in the natural environment has been a growing issue. This presence could also account for the influence that affects microorganisms in such a way that they develop resistance against these antibiotics. The aim of this study was to evaluate whether the antibiotic resistant gene (ARG) plasmid transfer can be facilitated by the impact of 1) environmentally representative micro-contaminant concentrations in ppb (part per billion) levels and 2) donor-recipient microbial complexity (pure vs. mixed). For this purpose, the multidrug resistant plasmid, pB10, and Escherichia coli DH5α were used as a model plasmid and a model donor, respectively. Based on conjugation experiments with pure (Pseudomonas aeruginosa PAKexoT) and mixed (activated sludge) cultures as recipients, increased relative plasmid transfer frequencies were observed at ppb (µg/L) levels of tetracycline and sulfamethoxazole micro-contaminant exposure. When sludge, a more complex community, was used as a recipient, the increases of the plasmid transfer rate were always statistically significant but not always in P. aeruginosa. The low concentration (10 ppb) of tetracycline exposure led to the pB10 transfer to enteric bacteria, which are clinically important pathogens.

Nitrate and ammonium ions removal from groundwater by a hybrid system of zero-valent iron combined with adsorbents.

Nitrate (NO(3)(-)) is a commonly found contaminant in groundwater and surface water. It has created a major water quality problem worldwide. The laboratory batch experiments were conducted to investigate the feasibility of HCl-treated zero-valent iron (Fe(0)) combined with different adsorbents as hybrid systems for simultaneous removal of nitrate (NO(3)(-)) and ammonium (NH(4)(+)) ions from aqueous solution. The maximum NO(3)(-) removal in combined Fe(0)-granular activated carbon (GAC), Fe(0)-filtralite and Fe(0)-sepiolite systems was 86, 96 and 99%, respectively, at 45 °C for 24 h reaction time. The NO(3)(-) removal rate increased with the increase in initial NO(3)(-) concentration. The NO(3)(-) removal efficiency by hybrid systems was in the order of sepiolite > filtralite > GAC. The NH(4)(+) produced during the denitrification process by Fe(0) was successfully removed by the adsorbents, with the removal efficiency in the order of GAC > sepiolite > filtralite. Results of the present study suggest that the use of a hybrid system could be a promising technology for achieving simultaneous removal of NO(3)(-) and NH(4)(+) ions from aqueous solution.

Removal of nitrate and ammonium ions from livestock wastewater by hybrid systems composed of zero-valent iron and adsorbents.

The feasibility of hybrid systems for simultaneous removal of nitrate (NO3-) and ammonium ions (NH4+) from livestock wastewater was examined in batch experiments. As a part of efforts to remove nitrate and ammonium simultaneously, Fe0 and adsorbents including coconut-based granular activated carbon (GAC), sepiolite and filtralite were used. Various parameters such as adsorbent dosages and temperature were studied. Removal of NO3- increased with increase in temperature. Maximum NO3- removal (85.3%) was observed for the Fe0-filtralite hybrid system at 45 degrees C for a 24 h reaction time. Increase in GAC and sepiolite dosages had significant (P < 0.01) effect on the NH4+ removal efficiency, which was primarily due to the net negative surface charge of the adsorbents. The efficiency of hybrid systems for the removal of NO3- was in the order of filtralite > sepiolite > GAC, and the order of the removal of NH4+ was GAC > sepiolite > filtralite. The results of the present study suggest that the use of hybrid systems could be a promising innovative technology for achieving simultaneous removal of NO3- and NH4 from livestock wastewater.

Degradation of PCE, TCE and 1,1,1-TCA by nanosized FePd bimetallic particles under various experimental conditions.

The degradation of chlorinated organic compounds, such as PCE (tetrachloroethene), TCE (trichloroethene) and 1,1,1-TCA (1,1,1-trichloroethane), was conducted using nanosized FePd bimetallic particles. In order to enhance the reactivity of ZVI (zero valent iron) nanoparticles, surface modification of ZVI nanoparticles was performed using Pd and CMC (carboxymethyl cellulose). The surface modification was found to form CMC-stabilized FePd bimetallic nanoparticles (CMC-FePd). The average TCE removal efficiency by the CMC-FePd was significantly increased by ∼85% compared to employing conventional ZVI nanoparticles (∼15%). This increase in the TCE removal efficiency was most likely due to the increased amount of atomic hydrogen produced by the formation of CMC-FePd. For PCE and 1,1,1-TCA, the removal efficiencies by CMC-FePd were approximately 80% and 56%, respectively. For all three chlorinated organic compounds, the amount of Cl- ions in the aqueous phase during the degradation increased with increasing reaction time. This result suggests that the main degradation mechanism of the chlorinated compounds by CMC-FePd was reductive dechlorination.