CHARACTERIZATION OF MICROBIAL COMMUNITY STRUCTURE A FTER APPLICATION OF DIFFERENT BIOREMEDIATION APPROACHES IN TNT CONTAMINATED SOIL

Contamination of the environment with explosive res idues presents a serious problem at sites across th e world. Trinitrotoluene (TNT) is one of the most com monly used explosive for military and industrial ap plications. In this study, bioaugmentation, biostimulation and phy toremediation were used as bioremediation strategie s. Effect of the higher plants (rye Secale cereale and blue fenugreek Trigonella caerulea ), amendments and specific bacteria consortium was studied in soil spiked with 118mg TNT/kg. Diversity of microorganisms and fate of TNT were evaluated after application of different bioremedia tion approaches. Results of Biolog Ecoplates data analysis showed that intensity of substrate assimilation by soil microb ial community was altered by application of the consortium. The i mpact of vegetation on the microbial community metabolic pro- files was also demonstrated. Shannon diversity inde x values obtained after 48 hours Biolog EcoPlates in cubation, was the highest in the samples with rye and fenugreek c ultivation. In unplanted soil samples, the Shannon diversity index rose when consortium of bacteria, nitroaromatic compounds and amendments were added. Inoculation of soil samples with mixture of bacterial isolates had effect on mi crobial community composition revealed by 16S rDNA-DGGE analysis. However, no clear effect of the vegetatio n on the microbial community structure was found according to DGGE results. The most pronounced effect of bioaugmentation and biostimulation for TNT degradation was shown in the variants with rye cultivation. At the same t ime, the use of qPCR method allowed to detect the most profound effect of biostimulation and bioaugmentation on soi l microbiological parameters in case of blue fenugr eek application.

[1]  N. Bruce,et al.  Plants disarm soil: engineering plants for the phytoremediation of explosives. , 2009, Trends in biotechnology.

[2]  B. Glick,et al.  Phytoremediation and rhizoremediation of organic soil contaminants : Potential and challenges , 2009 .

[3]  B. Van Aken.  Transgenic plants for enhanced phytoremediation of toxic explosives. , 2009, Current opinion in biotechnology.

[4]  O. Muter,et al.  Effect of plant extract on the degradation of nitroaromatic compounds by soil microorganisms , 2008, Journal of Industrial Microbiology & Biotechnology.

[5]  E. Travis,et al.  Microbial and plant ecology of a long-term TNT-contaminated site. , 2008, Environmental pollution.

[6]  E. Travis,et al.  Short term exposure to elevated trinitrotoluene concentrations induced structural and functional changes in the soil bacterial community. , 2008, Environmental pollution.

[7]  E. Travis,et al.  Impact of transgenic tobacco on trinitrotoluene (TNT) contaminated soil community. , 2007, Environmental science & technology.

[8]  J. Ramos,et al.  Bioremediation of 2,4,6-trinitrotoluene under field conditions. , 2007, Environmental science & technology.

[9]  R. B. Jackson,et al.  Assessment of Soil Microbial Community Structure by Use of Taxon-Specific Quantitative PCR Assays , 2005, Applied and Environmental Microbiology.

[10]  N. Scrutton,et al.  Biotransformation of Explosives by the Old Yellow Enzyme Family of Flavoproteins , 2004, Applied and Environmental Microbiology.

[11]  R. Crawford,et al.  Bioremediation of soils contaminated with explosives. , 2004, Journal of environmental management.

[12]  Xiaohan Yang,et al.  The role of root exudates and allelochemicals in the rhizosphere , 2003, Plant and Soil.

[13]  Peter J. Gregory,et al.  Changes in phosphorus concentrations and pH in the rhizosphere of some agroforestry and crop species , 2002, Plant and Soil.

[14]  E. Lagendijk,et al.  Rhizoremediation: a beneficial plant-microbe interaction. , 2004, Molecular plant-microbe interactions : MPMI.

[15]  M. Drew,et al.  The use of vegetation to remediate soil freshly contaminated by recalcitrant contaminants. , 2003, Water research.

[16]  R. Conrad,et al.  Quantitative Detection of Methanotrophs in Soil by Novel pmoA-Targeted Real-Time PCR Assays , 2003, Applied and Environmental Microbiology.

[17]  Dick B Janssen,et al.  Efficient recovery of environmental DNA for expression cloning by indirect extraction methods. , 2003, FEMS microbiology ecology.

[18]  Chulhwan Park,et al.  Optimization for biodegradation of 2,4,6-trinitrotoluene (TNT) by Pseudomonas putida. , 2003, Journal of bioscience and bioengineering.

[19]  N. Bruce,et al.  Phytoremediation of Explosives , 2002 .

[20]  J. Ramos,et al.  Biological Degradation of 2,4,6-Trinitrotoluene , 2001, Microbiology and Molecular Biology Reviews.

[21]  S. Sørensen,et al.  Effects of mercury contamination on the culturable heterotrophic, functional and genetic diversity of the bacterial community in soil. , 2001, FEMS microbiology ecology.

[22]  Judith C. Pennington,et al.  Monitored Natural Attenuation of Explosives , 2001 .

[23]  E. Travis,et al.  Microbial transformations of explosives. , 2001, Advances in applied microbiology.

[24]  S. Siciliano,et al.  Assessment of 2,4,6‐trinitrotoluene toxicity in field soils by pollution‐induced community tolerance, denaturing gradient gel electrophoresis, and seed germination assay , 2000 .

[25]  R. Drijber,et al.  Fate of 2,4,6‐trinitrotoluene in axenic sand culture systems containing smooth bromegrass , 2000 .

[26]  R. Meagher,et al.  Phytoremediation of toxic elemental and organic pollutants. , 2000, Current opinion in plant biology.

[27]  W. L. Powers,et al.  Growth and development of smooth bromegrass and tall fescue in TNT-contaminated soil. , 2000, Environmental pollution.

[28]  N. Panikov.  Understanding and prediction of soil microbial community dynamics under global change , 1999 .

[29]  B. Wilke,et al.  Soil-Based Phytotoxicity of 2,4,6-Trinitrotoluene (TNT) to Terrestrial Higher Plants , 1999, Archives of environmental contamination and toxicology.

[30]  M. Fuller,et al.  Evidence for differential effects of 2,4,6‐trinitrotoluene and other munitions compounds on specific subpopulations of soil microbial communities , 1998 .

[31]  S. Comfort,et al.  Germination and seedling development of switchgrass and smooth bromegrass exposed to 2,4,6-trinitrotoluene. , 1998, Environmental pollution.

[32]  J. Garland.  Analysis and interpretation of community-level physiological profiles in microbial ecology , 1997 .

[33]  A. J. Stewart,et al.  Multispecies toxicity assessment of compost produced in bioremediation of an explosives‐contaminated sediment , 1997 .

[34]  Charolett A. Hayes,et al.  Effect of Redox Potential and pH on TNT Transformation in Soil-Water Slurries , 1997 .

[35]  L. Forney,et al.  Distribution of bacterioplankton in meromictic Lake Saelenvannet, as determined by denaturing gradient gel electrophoresis of PCR-amplified gene fragments coding for 16S rRNA , 1997, Applied and environmental microbiology.

[36]  M. Fuller,et al.  Aerobic Gram-Positive and Gram-Negative Bacteria Exhibit Differential Sensitivity to and Transformation of 2,4,6-Trinitrotoluene (TNT) , 1997, Current Microbiology.

[37]  A. Uitterlinden,et al.  Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA , 1993, Applied and environmental microbiology.

[38]  Chen.  Effects of m , 1992, Physical review letters.

[39]  V. Sheffield,et al.  Attachment of a 40-base-pair G + C-rich sequence (GC-clamp) to genomic DNA fragments by the polymerase chain reaction results in improved detection of single-base changes. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[40]  Stefan Nowak,et al.  Understanding and prediction , 1976 .