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A new P. putida instrumental toxicity bioassay

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Abstract

Here, we present a new toxicity bioassay (CO2-TOX), able to detect toxic or inhibitory compounds in water samples, based on the quantification of Pseudomonas putida KT2440 CO2 production. The metabolically produced CO2 was measured continuously and directly in the liquid assay media, with a potentiometric gas electrode. The optimization studies were performed using as a model toxicant 3,5-DCP (3,5-dichlorophenol); later, heavy metals (Pb2+, Cu2+, or Zn2+) and a metalloid (As5+) were assayed. The response to toxics was evident after 15 min of incubation and at relatively low concentrations (e.g., 1.1 mg/L of 3,5-DCP), showing that the CO2-TOX bioassay is fast and sensitive. The EC50 values obtained were 4.93, 0.12, 6.05, 32.17, and 37.81 mg/L for 3,5-DCP, Cu2+, Zn2+, As5+, and Pb2+, respectively, at neutral pH. Additionally, the effect of the pH of the sample and the use of lyophilized bacteria were also analyzed showing that the bioassay can be implemented in different conditions. Moreover, highly turbid samples and samples with very low oxygen levels were measured successfully with the new instrumental bioassay described here. Finally, simulated samples containing 3,5-DCP or a heavy metal mixture were tested using the proposed bioassay and a standard ISO bioassay, showing that our test is more sensible to the phenol but less sensible to the metal mixtures. Therefore, we propose CO2-TOX as a rapid, sensitive, low-cost, and robust instrumental bioassay that could perform as an industrial wastewater-process monitor among other applications.

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Acknowledgments

This project was funded by the Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET) and the Agencia Nacional de Promoción Científica y Tecnológica (AGENCIA); we also want to acknowledge the support of the Subsecretaría de Recursos Hídricos de la Nación (Dr. Andrés Rodríguez), y la Dirección Ejecutiva de la Comisión Trinacional para el Desarrollo de la Cuenca del río Pilcomayo (Ing. Claudio Laboranti). We want to thank Ms. Silvia Rodriguez for the English edition.

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Authors and Affiliations

  1. Laboratory of Biosensors and Bioanalysis (LABB), Departamento de Química Biológica e IQUIBICEN-CONICET, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Pabellón 2, Ciudad Universitaria, Ciudad Autónoma de Buenos Aires, 1428, Argentina

    Federico Figueredo & Eduardo Cortón

  2. Instituto de Astronomía y Física del Espacio, CONICET–UBA, Pabellón IAFE, Ciudad Universitaria, Ciudad Autónoma de Buenos Aires, 1428, Argentina

    Ximena C. Abrevaya

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  1. Federico Figueredo
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  2. Ximena C. Abrevaya
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  3. Eduardo Cortón
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Correspondence to Eduardo Cortón.

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Figueredo, F., Abrevaya, X.C. & Cortón, E. A new P. putida instrumental toxicity bioassay. Environ Monit Assess 187, 294 (2015). https://doi.org/10.1007/s10661-015-4499-1

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