Influence of phenolics on the sensitivity of free and immobilized bioluminescent Acinetobacter bacterium.

In this work, the constructed bioluminescent Acinetobacter strain DF4/PUTK2 was employed to assess the toxicity of phenolic compounds and the 5 min EC50 values were calculated. The results of the DF4/PUTK2 assay were further evaluated by comparing with the results of the Vibrio fischeri luminescence inhibition assay. To develop a bioassay system appropriate to be used in continuous toxicity testing, strain DF4/PUTK2 was subjected for immobilization in microtiter plates into the matrices Ca-alginate, polyacrylamide, agar and agarose. After a choice of materials was tried, Ca-alginate was selected as the most suitable candidate material. Because, it could be stored at least 8 weeks at 4 degrees C, during which the ability of the bioreporter DF4/PUTK2 to detect the toxicity of phenolics was maintained. However, the stability of the bioluminescence for DF4/PUTK2 cells immobilized into agarose and agar was significantly less than that of cells stored in alginate suspensions. This study recommended that luxCDABE-marked Acinetobacter strain DF4/PUTK2 could be employed to assay the ecotoxicity of environmental samples contaminated with phenols. The host strain of the bioreporter DF4/PUTK2 is Acinetobacter strain DF4. It is known that members of the genus Acinetobacter are widespread in nature and also involved in biodegradation, leaching and removal of several organic and inorganic man-made hazardous wastes.

Acinetobacter bioreporter assessing heavy metals toxicity.

This work was conducted to employ a whole cell-based biosensor to monitor toxicity of heavy metals in water and wastewater. An isolate of industrial wastewater bacterium, Acinetobacter sp. DF4, was genetically modified with lux reporter gene to create a novel bioluminescent bacterial strain, designated as DF4/PUTK2. This bioreporter can investigate the toxicity through light inhibition due to cell death or metabolic burden and the specific stress effects of the tested soluble materials simultaneously. The use of Acinetobacter DF4/PUTK2 as a bioluminescent reporter for heavy metal toxicity testing and for the application of wastewater treatment influent toxicity screening is presented in this study. Among eight heavy metals tested, the bioluminescence of DF4/PUTK2 was most sensitive to Zn, Cd, Fe, Co, Cr followed by Cu in order of decreasing sensitivity. The same pattern of sensitivity was observed when several contaminated water and wastewater effluents were assayed. This work suggested that luxCDABE -marked Acinetobacter bacterium DF4/PUTK2 can be used to bioassay the ecotoxicity of wastewater and effluent samples contaminated with heavy metals. Using this assay, it is possible to pre-select the more toxic samples for further chemical analysis and to discard wastewater samples with low or no inhibition because they are not toxic to the environment.