Hydrocarbon biodegradation and dynamic laser speckle for detecting chemotactic responses at low bacterial concentration.

We report on the biodegradation of pure hydrocarbons and chemotaxis towards these compounds by an isolated chlorophenol degrader, Pseudomonas strain H. The biochemical and phylogenetic analysis of the 16S rDNA sequence identified Pseudomonas strain H as having 99.56% similarity with P. aeruginosa PA01. This strain was able to degrade n-hexadecane, 1-undecene, 1-nonene, 1-decene, 1-dodecene and kerosene. It grew in the presence of 1-octene, while this hydrocarbons is toxic to other hydrocarbons degraders. Pseudomonas strain H was also chemotactic towards n-hexadecane, kerosene, 1-undecene and 1-dodecene. These results show that this Pseudomonas strain H is an attractive candidate for hydrocarbon-containing wastewater bioremediation in controlled environments. Since the classical standard techniques for detecting chemotaxis are not efficient at low bacterial concentrations, we demonstrate the use of the dynamic speckle laser method, which is simple and inexpensive, to confirm bacterial chemotaxis at low cell concentrations (less than 10(5) colony-forming unit per millilitre (CFU/mL)) when hydrocarbons are the attractants.

[1]  Lin Aijun,et al.  Bio-removal of mixture of benzene, toluene, ethylbenzene, and xylenes/total petroleum hydrocarbons/trichloroethylene from contaminated water. , 2009 .

[2]  H. Shim,et al.  Bio-removal of mixture of benzene, toluene, ethylbenzene, and xylenes/total petroleum hydrocarbons/trichloroethylene from contaminated water. , 2009, Journal of Environmental Science.

[3]  C. Studdert,et al.  A strain isolated from gas oil-contaminated soil displays chemotaxis towards gas oil and hexadecane. , 2003, Environmental microbiology.

[4]  M. R. Bruins,et al.  Pseudomonas pickettii: a common soil and groundwater aerobic bacteria with pathogenic and biodegradation properties. , 2000, Ecotoxicology and environmental safety.

[5]  S. Panke,et al.  Analysis of Pseudomonas putida alkane-degradation gene clusters and flanking insertion sequences: evolution and regulation of the alk genes. , 2001, Microbiology.

[6]  J. Thompson,et al.  The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. , 1997, Nucleic acids research.

[7]  Q. Meng,et al.  Biodegradation of crude oil by Pseudomonas aeruginosa in the presence of rhamnolipids. , 2005, Journal of Zhejiang University. Science. B.

[8]  M. Ilori,et al.  Differential degradation of crude oil (Bonny Light) by four Pseudomonas strains. , 2009, Journal of environmental sciences.

[9]  C. Harwood,et al.  Bacterial chemotaxis to pollutants and plant-derived aromatic molecules. , 2002, Current opinion in microbiology.

[10]  Sang Moo Kim,et al.  Agarase: Review of Major Sources, Categories, Purification Method, Enzyme Characteristics and Applications , 2010, Marine drugs.

[11]  Kevin C Jones,et al.  Defining bioavailability and bioaccessibility of contaminated soil and sediment is complicated. , 2004, Environmental science & technology.

[12]  J. Wong,et al.  Biosurfactants from Acinetobacter calcoaceticus BU03 enhance the solubility and biodegradation of phenanthrene , 2009, Environmental technology.

[13]  N. Olivera,et al.  Microbial characterization and hydrocarbon biodegradation potential of natural bilge waste microflora , 2003, Journal of Industrial Microbiology and Biotechnology.

[14]  A. Mukherjee,et al.  Crude petroleum-oil biodegradation efficiency of Bacillus subtilis and Pseudomonas aeruginosa strains isolated from a petroleum-oil contaminated soil from North-East India. , 2007, Bioresource technology.

[15]  E. Okada,et al.  Bacillus subtilis as a bioindicator for estimating pentachlorophenol toxicity and concentration , 2009, Journal of Industrial Microbiology & Biotechnology.

[16]  M Trivi,et al.  Decomposition of biospeckle images in temporary spectral bands. , 2005, Optics letters.

[17]  R. Colwell,et al.  Microbial degradation of hydrocarbons in the environment. , 1990, Microbiological reviews.

[18]  J. Pandey,et al.  Accessing microbial diversity for bioremediation and environmental restoration. , 2005, Trends in biotechnology.

[19]  J B Armstrong,et al.  Nonchemotactic Mutants of Escherichia coli , 1967, Journal of bacteriology.

[20]  J. Trevors,et al.  A drop-collapsing test for screening surfactant-producing microorganisms , 1991 .

[21]  J. González,et al.  Degradation of phenol and chlorophenols by mixed and pure cultures , 2004 .

[22]  Ann C. Grimm,et al.  Chemotaxis of Pseudomonas spp. to the polyaromatic hydrocarbon naphthalene , 1997, Applied and environmental microbiology.

[23]  J. González,et al.  Degradation of chlorophenol mixtures in a fed-batch system by two soil bacteria , 2011 .

[24]  J. Lalucat,et al.  Identification of culturable bacteria present in haemodialysis water and fluid. , 2005, FEMS microbiology ecology.

[25]  Fritz Wagner,et al.  New method for detecting rhamnolipids excreted by Pseudomonas species during growth on mineral agar , 1991 .

[26]  A. K. Haritash,et al.  Biodegradation aspects of polycyclic aromatic hydrocarbons (PAHs): a review. , 2009, Journal of hazardous materials.

[27]  K. Koutsoumanis,et al.  Comparison of maximum specific growth rates and lag times estimated from absorbance and viable count data by different mathematical models. , 2001, Journal of microbiological methods.

[28]  M. Malmqvist Purification and characterization of two different agarose-degrading enzymes. , 1978, Biochimica et biophysica acta.

[29]  J. Ortega-Calvo,et al.  Chemotaxis in polycyclic aromatic hydrocarbon-degrading bacteria isolated from coal-tar- and oil-polluted rhizospheres. , 2003, FEMS microbiology ecology.

[30]  Ignacio Durruty,et al.  Multisubstrate monod kinetic model for simultaneous degradation of chlorophenol mixtures , 2011 .

[31]  H. Heipieper,et al.  Mechanisms of resistance of whole cells to toxic organic solvents , 1994 .

[32]  Alicja Szulc,et al.  Interactions between rhamnolipid biosurfactants and toxic chlorinated phenols enhance biodegradation of a model hydrocarbon-rich effluent , 2011 .

[33]  Jeanne Mager Stellman,et al.  Encyclopaedia of occupational health and safety , 1998 .

[34]  Héctor Rabal,et al.  Analysis of bacterial chemotactic response using dynamic laser speckle. , 2009, Journal of biomedical optics.

[35]  Hauke Harms,et al.  Principles of microbial PAH-degradation in soil. , 2005, Environmental pollution.

[36]  J. S. Parkinson A "bucket of light" for viewing bacterial colonies in soft agar. , 2007, Methods in enzymology.

[37]  J. González,et al.  Effect of pH and inoculum size on pentachlorophenol degradation by Pseudomonas sp. , 2007 .

[38]  K. Furukawa 'Super bugs' for bioremediation. , 2003, Trends in biotechnology.

[39]  J. Mueller,et al.  Biodegradation of creosote and pentachlorophenol in contaminated groundwater: chemical and biological assessment , 1991, Applied and environmental microbiology.

[40]  H. Berg,et al.  Migration of bacteria in semisolid agar. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[41]  Q. Shen,et al.  Novel rhamnolipid biosurfactants produced by a polycyclic aromatic hydrocarbon-degrading bacterium Pseudomonas aeruginosa strain NY3. , 2010, Biotechnology advances.

[42]  A. Johnsen,et al.  Detection of Microbial Growth on Polycyclic Aromatic Hydrocarbons in Microtiter Plates by Using the Respiration Indicator WST-1 , 2002, Applied and Environmental Microbiology.

[43]  L. M. Lahner,et al.  Degradation of straight-chain aliphatic and high-molecular-weight polycyclic aromatic hydrocarbons by a strain of Mycobacterium austroafricanum. , 2003, Journal of applied microbiology.

[44]  J. L. Ditty,et al.  Toluene-Degrading Bacteria Are Chemotactic towards the Environmental Pollutants Benzene, Toluene, and Trichloroethylene , 2000, Applied and Environmental Microbiology.

[45]  D. Paul,et al.  Chemotaxis of Ralstonia sp. SJ98 towards p-nitrophenol in soil. , 2006, Environmental microbiology.

[46]  R. Maier,et al.  Pseudomonas aeruginosa rhamnolipids: biosynthesis and potential applications , 2000, Applied Microbiology and Biotechnology.

[47]  N. Das,et al.  Microbial Degradation of Petroleum Hydrocarbon Contaminants: An Overview , 2010, Biotechnology research international.

[48]  M. D. Aitken,et al.  Bacterial Chemotaxis Enhances Naphthalene Degradation in a Heterogeneous Aqueous System , 2000 .

[49]  T. Phelps,et al.  Determining chemotactic responses by two subsurface microaerophiles using a simplified capillary assay method. , 1999, Journal of microbiological methods.