Bacterial communities in batch and continuous-flow wetlands treating the herbicide S-metolachlor.

Knowledge of wetland bacterial communities in the context of pesticide contamination and hydrological regime is scarce. We investigated the bacterial composition in constructed wetlands receiving Mercantor Gold(®) contaminated water (960 g L(-1) of the herbicide S-metolachlor, >80% of the S-enantiomer) operated under continuous-flow or batch modes to evaluate the impact of the hydraulic regime. In the continuous-flow wetland, S-metolachlor mass removal was >40%, whereas in the batch wetland, almost complete removal of S-metolachlor (93-97%) was observed. Detection of ethanesulfonic and oxanilic acid degradation products further indicated S-metolachlor biodegradation in the two wetlands. The dominant bacterial populations were characterised by terminal restriction fragment length polymorphism (T-RFLP) and 454 pyrosequencing. The bacterial profiles evolved during the first 35 days of the experiment, starting from a composition similar to that of inlet water, with the use of nitrate and to a lesser extent sulphate and manganese as terminal electron acceptors for microbial metabolism. Proteobacteria were the most abundant phylum, with Beta-, Alpha- and Gammaproteobacteria representing 26%, 19% and 17% respectively of total bacterial abundance. Bacterial composition in wetland water changed gradually over time in continuous-flow wetland and more abruptly in the batch wetland. Differences in overall bacterial water structure in the two systems were modest but significant (p=0.008), and S-metolachlor, nitrate, and total inorganic carbon concentrations correlated with changes in the bacterial profiles. Together, the results highlight that bacterial composition profiles and their dynamics may be used as bioindicators of herbicide exposure and hydraulic disturbances in wetland systems.

Using compound-specific isotope analysis to assess the degradation of chloroacetanilide herbicides in lab-scale wetlands.

Compound-specific isotope analysis (CSIA) is a promising tool to study the environmental fate of a wide range of contaminants including pesticides. In this study, a novel CSIA method was developed to analyse the stable carbon isotope signatures of widely used chloroacetanilide herbicides. The developed method was applied in combination with herbicide concentration and hydrochemical analyses to investigate in situ biodegradation of metolachlor, acetochlor and alachlor during their transport in lab-scale wetlands. Two distinct redox zones were identified in the wetlands. Oxic conditions prevailed close to the inlet of the four wetlands (oxygen concentration of 212±24µM), and anoxic conditions (oxygen concentrations of 28±41µM) prevailed towards the outlet, where dissipation of herbicides mainly occurred. Removal of acetochlor and alachlor from inlet to outlet of wetlands was 56% and 51%, whereas metolachlor was more persistent (23% of load dissipation). CSIA of chloroacetanilides at the inlet and outlet of the wetlands revealed carbon isotope fractionation of alachlor (εbulk=-2.0±0.3‰) and acetochlor (εbulk=-3.4±0.5‰), indicating that biodegradation contributes to the dissipation of both herbicides. This study is a first step towards the application of CSIA to evaluate the transport and degradation of chloroacetanilide herbicides in the environment.

Functional characterization of Fospeg, and its impact on cell cycle upon PDT of Huh7 hepatocellular carcinoma cell model.

BACKGROUND: Although several treatment options are available for hepatocellular carcinoma (HCC), their application is mostly restricted to early diagnosed cases or includes liver transplantation, which is rarely available due to donor scarcity. The attractiveness of PDT as a cancer treatment does not only come from its minimal invasiveness, but also from the high selectivity due to tumor localization that can be applied. Precise focusing of light on tumor lesions will result in tumor-specific PDT activation. Novel photosensitizers can be applied in such low concentrations that cells not subjected to irradiation remain healthy. The lethal effect and mechanism of death induction of the photosensitizer Fospeg has never been studied on hepatocellular carcinoma. The aim of the present study is to functionally analyze the impact of PDT on Huh-7 HCC cell line, as well as to analyze its impact on cell cycle protein expression. METHODS: Cellular viability, and proliferation assays were conducted via MTT and BrdU assay, respectively. Transfected cell models of Huh7 with different constructs harboring cell cycle genes and downstream reporter luciferase gene were generated. RESULTS: Our results show a statistically significant decrease in both viability and proliferation of Huh-7 cells following PDT, while maintaining Fospeg and laser concentrations far below toxic levels. Proliferative cell cycle genes show a tendency of inhibition, while p53 levels show a significant increase following PDT. CONCLUSION: Fospeg-mediated PDT is a promising strategy for treatment of hepatocellular carcinoma and needs to be further explored in vivo.