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Bioremediation Approaches in a Laboratory Activity for the Industrial Biotechnology and Applied Microbiology (IBAM) Course

    Authors: L. Raiger Iustman1, N. I. López1, S. M. Ruzal1, D. L. Vullo1,2,*
    VIEW AFFILIATIONS HIDE AFFILIATIONS
    Affiliations: 1: Área Microbiología, Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, (1428) Buenos Aires, Argentina; 2: Área Química, Instituto de Ciencias, Universidad Nacional General Sarmiento-CONICET, (B1613GSX) Buenos Aires, Argentina
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • Published 06 May 2013
    • Supplemental materials available at http://jmbe.asm.org
    • *Corresponding author. Mailing address: Área Química, Instituto de Ciencias, Universidad Nacional General Sarmiento, J.M. Gutierrez 1150, (B1613GSX) Los Polvorines, Buenos Aires, Argentina. Phone: 5411-4469-7542. Fax: 5411-4469-7506. E-mail: dvullo@ungs.edu.ar.
    • ©2013 Author(s). Published by the American Society for Microbiology.
    Source: J. Microbiol. Biol. Educ. May 2013 vol. 14 no. 1 131-134. doi:10.1128/jmbe.v14i1.558
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    Abstract:

    Industrial Biotechnology and Applied Microbiology is an optional 128h-course for Chemistry and Biology students at the Faculty of Sciences, University of Buenos Aires, Argentina. This course is usually attended by 25 students, working in teams of two. The curriculum, with 8 lab exercises, includes an oil bioremediation practice covering an insight of bioremediation processes: the influence of pollutants on autochthonous microbiota, biodegrader isolation and biosurfactant production for bioavailability understanding. The experimental steps are: (A) evaluation of microbial tolerance to pollutants by constructing pristine soil microcosms contaminated with diesel or xylene and (B) isolation of degraders and biosurfactant production analysis. To check microbial tolerance, microcosms are incubated during one week at 25-28ºC. Samples are collected at 0, 4 and every 48 h for CFU/g soil testing. An initial decrease of total CFU/g related to toxicity is noticed. At the end of the experiment, a recovery of the CFU number is observed, evidencing enrichment in biodegraders. Some colonies from the CFU counting plates are streaked in M9-agar with diesel as sole carbon source. After a week, isolates are inoculated on M9-Broth supplemented with diesel to induce biosurfactant production. Surface tension and Emulsification Index are measured in culture supernatants to visualize tensioactive effect of bacterial products. Besides the improvement in the good microbiological practices, the students show enthusiasm in different aspects, depending on their own interests. While biology students explore and learn new concepts on solubility, emulsions and bioavailability, chemistry students show curiosity in bacterial behavior and manipulation of microorganisms for environmental benefits.

Key Concept Ranking

Applied and Industrial Microbiology
0.97222227
Soil Microbial Communities
0.43604717
Pseudomonas putida
0.4074074
0.97222227

References & Citations

1. Atlas RM, Bartha R 2001 Ecología microbiana y microbiología ambiental 4th ed. Pearson Educación Madrid, Spain
2. Di Martino C, López NI, Raiger Iustman LJ 2012 Isolation and characterization of benzene, toluene, and xylene degrading Pseudomonas spp. selected as candidates for bioremediation Int. Biodeterior. Biodegradation 67 15 20 10.1016/j.ibiod.2011.11.004 http://dx.doi.org/10.1016/j.ibiod.2011.11.004
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/content/journal/jmbe/10.1128/jmbe.v14i1.558
2013-05-06
2017-11-19

Abstract:

Industrial Biotechnology and Applied Microbiology is an optional 128h-course for Chemistry and Biology students at the Faculty of Sciences, University of Buenos Aires, Argentina. This course is usually attended by 25 students, working in teams of two. The curriculum, with 8 lab exercises, includes an oil bioremediation practice covering an insight of bioremediation processes: the influence of pollutants on autochthonous microbiota, biodegrader isolation and biosurfactant production for bioavailability understanding. The experimental steps are: (A) evaluation of microbial tolerance to pollutants by constructing pristine soil microcosms contaminated with diesel or xylene and (B) isolation of degraders and biosurfactant production analysis. To check microbial tolerance, microcosms are incubated during one week at 25-28ºC. Samples are collected at 0, 4 and every 48 h for CFU/g soil testing. An initial decrease of total CFU/g related to toxicity is noticed. At the end of the experiment, a recovery of the CFU number is observed, evidencing enrichment in biodegraders. Some colonies from the CFU counting plates are streaked in M9-agar with diesel as sole carbon source. After a week, isolates are inoculated on M9-Broth supplemented with diesel to induce biosurfactant production. Surface tension and Emulsification Index are measured in culture supernatants to visualize tensioactive effect of bacterial products. Besides the improvement in the good microbiological practices, the students show enthusiasm in different aspects, depending on their own interests. While biology students explore and learn new concepts on solubility, emulsions and bioavailability, chemistry students show curiosity in bacterial behavior and manipulation of microorganisms for environmental benefits.

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Figures

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FIGURE 1

Diagram of the inoculation pattern for diesel degrader selection in plate with small channels.

Source: J. Microbiol. Biol. Educ. May 2013 vol. 14 no. 1 131-134. doi:10.1128/jmbe.v14i1.558
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Image of FIGURE 2

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FIGURE 2

Du Nouy tensiometer.

Source: J. Microbiol. Biol. Educ. May 2013 vol. 14 no. 1 131-134. doi:10.1128/jmbe.v14i1.558
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Image of FIGURE 3

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FIGURE 3

(A) CFU/g soil in microcosms along incubation time at room temperature. (B) Increase of the morphological homogeneity of the colonies obtained after long exposures to contaminants.

Source: J. Microbiol. Biol. Educ. May 2013 vol. 14 no. 1 131-134. doi:10.1128/jmbe.v14i1.558
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FIGURE 4

Biosurfactant production of selected strains: EI analysis by calculating the quotient between emulsion height of liquid phases and the total height.

Source: J. Microbiol. Biol. Educ. May 2013 vol. 14 no. 1 131-134. doi:10.1128/jmbe.v14i1.558
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