RESUMO
Surfactants have a widespread occurrence, not only as household detergents, but also in their application in industry and medicine. There are numerous bioassays for assessing surfactant toxicity, but investigations of their impact on biological systems at the molecular level are still needed. In this paper, luminous marine bacteria and their coupled NAD(P)H:FMN-oxidoreductase + luciferase (Red + Luc) enzyme system was applied to examine the effects of different types of surfactants, including cationic cetyltrimethylammonium bromide (CTAB), non-ionic polyoxyethylene 20 sorbitan monooleate (Tween 80) and anionic sodium lauryl sulfate (SLS), and to assess whether the Red + Luc enzyme system can be used as a more sensitive indicator of toxicity. It was shown that the greatest inhibitory effect of the surfactants on the activity of luminous bacteria and the Red + Luc enzyme system was in the presence of SLS samples. The calculated IC50 and EC50 values of SLS were 10-5 M and 10-2 M for the enzymatic and cellular assay systems, respectively. The results highlight the benefits of using the enzymatic assay system in ecotoxicology as a tool for revealing surfactant effects on intracellular proteins if the cellular membrane is damaged under a long-term exposure period in the presence of the surfactants. For this purpose, the bioluminescent enzyme-inhibition-based assay could be used as an advanced research tool for the evaluation of surfactant toxicity at the molecular level of living organisms due to its technical simplicity and rapid response time.
Assuntos
Surfactantes Pulmonares , Tensoativos , Tensoativos/toxicidade , Dodecilsulfato de Sódio/toxicidade , Polissorbatos , CetrimônioRESUMO
This work is dedicated to developing enzyme biosensor software to solve problems regarding soil pollution analysis. An algorithm and specialised software have been developed which stores, analyses and visualises data using JavaScript programming language. The developed software is based on matching data of 51 non-commercial standard soil samples and their inhibitory effects on three enzyme systems of varying complexity. This approach is able to identify the influence of chemical properties soil samples, without toxic agents, on enzyme biosensors. Such software may find wide use in environmental monitoring.
Assuntos
Monitoramento Ambiental , Software , Poluentes do Solo , Técnicas Biossensoriais , Poluição Ambiental , Padrões de Referência , Solo , Microbiologia do Solo , Poluentes do Solo/análiseRESUMO
A bioluminescent-enzyme-inhibition-based assay was applied to predict the potential toxicity of the full profile of the following soil samples: agricultural grassland, 10-year fallow land (treated with remediation processes for 10 years) and uncontaminated (virgin) land. This assay specifically detects the influence of aqueous soil extracts from soils on the activity of a coupled enzyme system of luminescent bacteria: NAD(P)H:FMN-oxidoreductase + luciferase (Red + Luc). It was shown that the inhibitory effect of the full-profile soil samples on the Red + Luc system decreased with depth for the 10-year fallow-land and virgin-land samples, which correlated with a decrease in the humic organic matter content in the soils. The inhibitory effect of the agricultural grassland on the Red + Luc enzyme system activity was more complex and involved the presence of the humic organic matter content, as well as the presence of pollutants in the whole-soil profile. However, if the interfering effect of humic organic substances on the Red + Luc system's activity is taken into account during full-profile soil toxicity assessments, it might help to detect pollutant mobility and its leaching into the subsoil layer. Thus, this bioluminescent method, due to the technical simplicity, rapid response time and high sensitivity, has the potential to be developed as a biological part of the inhibition-based assay and/or biosensors for the preventive tracing of potential full-profile soil contamination.