Detection of microorganisms involved in metabolism of organic environmental pollutants

Abstract

A method and point of use kit to determine under field conditions the presence and/or amount of a selected Pseudomonas species capable of bioremediating organic environmental pollutants, such as petroleum hydrocarbons.

What is claimed is: 
     
         1 . A method to enhance detection of selected Pseudomonas microorganisms relative to other  Psuedomonas  microorganisms in water and soil samples, the method comprising
 placing a water and/or soil sample into a selective nutrient medium, and   incubating the medium to at least 20° C. but less than 37° C. for 24 h to 48 h,   visually evaluating without microscopy the incubated medium for presence of microbial colony formation, and   determining the presence and/or relative concentration of at least one of  Pseudomonas putida, Pseudomonas aeruginosa , or  Pseudomonas cepacia  microoganisms by formation of at least one colony on the medium.   
     
     
         2 . A method to enhance bioremediation of a water and/or soil sample potentially contaminated with organic environmental pollutants, the method comprising detecting the level of at least one  Pseudomonas  microorganism in the water and/or soil sample in the absence of microscopy identification and in the absence of incubating the microorganisms at a controlled temperature. 
     
     
         3 . The method of  claim 1  additionally determining at least one of  Pseudomonas stutzeri  or  Pseudomonas fluorescens  microorganisms. 
     
     
         4 . The method of  claim 2  where the  Pseudomonas  microorganisms are limited to  Pseudomonas putida, Pseudomonas aeruginosa, Pseudomonas cepacia, Pseudomonas stutzeri , and/or  Pseudomonas fluorescens.    
     
     
         5 . The method of  claim 1  where the selective growth medium comprises a peptic digest of animal tissue; magnesium chloride; potassium sulfate; erythromycin; ampicillin sodium salt; triclosan; tetrazolium blue chloride; agar; and optionally glycerol. 
     
     
         6 . The method of  claim 5  where the selective growth medium comprises a peptic digest of animal tissue 10 g/L-30 g/L; magnesium chloride 1 g/L-2 g/L; potassium sulfate 8 g/L-12 g/L; erythromycin 0.005 g/L-0.02 g/L; ampicillin sodium salt 0.005 g/L-0.02 g/L; triclosan 0.02 g/L-0.04 g/L; tetrazolium blue chloride 0.2 g/L-0.5 g/L; agar 12 g/L- 15  g/L; and optionally glycerol. 
     
     
         7 . A growth medium selective for growth of only  Pseudomonas putida, Pseudomonas aeruginosa, Pseudomonas cepacia, Pseudomonas stutzeri , and/or  Pseudomonas fluorescens , the medium comprising a peptic digest of animal tissue 10 g/L-30 g/L; magnesium chloride 1 g/L-2 g/L; potassium sulfate 8 g/L-12 g/L; erythromycin 0.005 g/L-0.02 g/L; ampicillin sodium salt 0.005 g/L-0.02 g/L; triclosan 0.02 g/L-0.04 g/L; tetrazolium blue chloride; agar 12 g/L-15 g/L; and excluding glycerol. 
     
     
         8 . A growth medium selective for growth of only  Pseudomonas putida, Pseudomonas aeruginosa , and  Pseudomonas cepacia , the media medium comprising a peptic digest of animal tissue 10 g/L-30 g/L; magnesium chloride 1 g/L-2 g/L; potassium sulfate 8 g/L-12 g/L; erythromycin 0.005 g/L-0.02 g/L; ampicillin sodium salt 0.005 g/L-0.02 g/L; triclosan 0.02 g/L-0.04 g/L; tetrazolium blue chloride 0.2 g/L-0.5 g/L; agar 12 g/L-15 g/L; and glycerol. 
     
     
         9 . A point of use kit to determine the presence or amount of a  Psuedomonas  species in a potentially contaminated water and/or soil sample, the kit comprising
 a container containing a nutrient media medium selective for growth limited to at least one  Pseudomonas  species selected from the group consisting of  Pseudomonas putida, Pseudomonas aeruginosa, Pseudomonas cepacia, Pseudomonas stutzeri, Pseudomonas fluorescens , and combinations thereof, and a dye or stain, and   instructions for assessing at least one of the  Psuedomonas  species in the potentially contaminated soil or water sample indicating potential for bioremediation of the water and/or soil sample.   
     
     
         10 . The kit of  claim 9  where the nutrient medium is a solid medium. 
     
     
         11 . The kit of  claim 9  where the nutrient medium is a liquid medium. 
     
     
         12 . The kit of  claim 9  further comprising an implement to manipulate the sample. 
     
     
         13 . A point-of-use kit to determine the presence or amount of a  Psuedomonas  species in a potentially contaminated water and/or soil sample, the point-of-use kit comprising
 a container containing the growth medium of  claim 8  selective for growth limited to at least one  Pseudomonas  species selected from the group consisting of  Pseudomonas putida, Pseudomonas aeruginosa, Pseudomonas cepacia, Pseudomonas stutzeri, Pseudomonas fluorescens , and combinations thereof, and a dye or stain, and   instructions for assessing at least one of the  Psuedomonas  species in the potentially contaminated water and/or soil sample indicating potential for bioremediation of the potentially contaminated water and/or soil sample.   
     
     
         14 . The point-of-use kit of  claim 13  where the nutrient medium is a solid medium. 
     
     
         15 . The point-of-use kit of  claim 13  where the nutrient medium is a liquid medium. 
     
     
         16 . The point-of-use kit of  claim 13  further comprising an implement to manipulate the potentially contaminated water and/or soil sample.