Microbiological oil prospecting

• US 5,093,236 A
• Filed: 06/22/1990
• Issued: 03/03/1992
• Est. Priority Date: 09/14/1988
• Status: Expired due to Term

• Current Assignee: Geo-Microbial Technologies Incorporated, Genencor International Incorporated (International Flavors & Fragrances Incorporated)
• Original Assignee: Geo-Microbial Technologies Incorporated, Genencor International Incorporated (International Flavors & Fragrances Incorporated)
• Inventors: Gonzales-Prevatt, Victoria, Munnecke, Douglas M.
• Primary Examiner(s): Rosen, Sam
• Application Number:
  US07/541,469
• Time in Patent Office: 620 Days
• Field of Search: 435/9
• US Class Current: 435/9
• CPC Class Codes: C12M 41/28 of redox potential
• CPC Class Codes: C12M 41/36 of biomass, e.g. colony counters or by turbidity measurements electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence
• CPC Class Codes: C12Q 1/64 Geomicrobiological testing, e.g. for petroleum
• CPC Class Codes: C12Q 2304/22 Resazurin; Resorufin
• CPC Class Codes: Y10S 435/81 Packaged device or kit

Abstract

This invention provides methods and systems for prospecting for subterranean oil and gas deposits by detecting the presence of hydrocarbon-consuming microorganisms in soil samples taken from a field of interest for production. A preferred method of this invention comprises suspending and diluting each soil sample in mineral salt solution, combining the sample with a selective growth, substrate such as an alcohol or aldehyde, and an oxidation-reduction dye in a gas-tight tube, incubating the sample and measuring the color change of the dye. Advantages of this system include: results obtained in short times (24 to 72 hours), substantially eliminates false positives, accurate results particularly when a spectrophotometer is used to measure the color changes, and it is automatable with computer controlled systems for processing samples, reading the color changes and calculating and plotting test results.

Claims

1. Method for microbiological prospecting for oil or gas comprising the steps of:

(i) suspending a soil sample in a mineral salt solution;

(ii) adding to a gas-tight tube;

(a) said suspended soil sample,

(b) an additional portion of said mineral salt solution,

(c) a selective electron-donating growth substrate capable of providing electron-donating potential for a hydrocarbon-consuming microorganism; and

(d) an oxidation-reduction dye capable of indicating by a change in color a change in the redox potential caused by growth of a hydrocarbon-consuming microorganism;

(iii) sealing the tube and incubating the contents of the sealed tube to allow growth of a hydrocarbon-consuming microorganism present therein; and

(iv) observing color change in the tube contents to determine the presence of a hydrocarbon-consuming microorganism in said soil sample.

2. A method according to claim 1 wherein the selective growth substrate comprises an alcohol or an aldehyde.

3. A method according to claim 1 wherein the dye comprises resazurin, methylene blue, or a tetrazolium dye.

4. A method according to claim 2 wherein the dye comprises resazurin, methylene blue , or a tetrazolium dye.

5. A method according to claim 1 wherein the color change is measured by spectrophotometric means and an automatable means compares the color change to predetermined color standards and calculates a quantitative indication of the presence of a hydrocarbon-consuming microorganism in the soil sample.

6. A system for detecting the presence of a hydrocarbon-consuming microorganism in a soil sample comprising:

(i) a gas-tight tube having resealable means for introducing a soil sample into the tube;

(ii) sealed within said tube;

(a) a mineral salt solution;

(b) a selective growth substrate capable of providing electron-donating potential to a hydrocarbon-consuming microorganism; and

(c) an oxidation-reduction dye capable of indicating by a change in color a change in the redox potential caused by growth of a hydracarbon-consuming microorganism in the tube.

7. A system according to claim 6 wherein a soil sample is dispersed in the mineral salt solution.

8. A system according to claim 6 wherein the selective growth substrate comprises an alcohol or an aldehyde.

9. A system according to claim 7 wherein the selective growth substrate comprises an alcohol or an aldehyde.

10. A system according to claim 8 wherein the dye comprises resazurin, methylene blue, or a tetrazolium dye. the dye comprises resazurin, methylene blue, or a tetrazolium dye

11. A system according to claim 9 wherein the dye comprises resazurin, methylene blue, or a tetrazolium dye.

12. A system according to claim 10 wherein the system further comprises spectrophotometric means for measuring a color change in the dye and automatable means for comparing the color change to a predetermined standard and calculating a quantitative indication of the presence of a hydrocarbon-consuming microorganism in the soil sample.

13. A system according to claim 11 wherein the system further comprises spectrophotometric means for measuring a color change in the dye and automa-table means for comparing the color change to a predetermined standard and calculating a quantitative indication of the presence of a hydrocarbon-consuming microorganism in the soil sample.

14. A kit for detecting the presence of hydrocarbon-consuming microorganisms in a soil sample comprising:

(i) a gas-tight tube having resealable opening means for introducing a soil sample into the tube; and

,

(ii) sealed within said tube;

(a) a soil sample suspended in a mineral salt solution;

(b) a selective growth substrate capable of providing electron-donating potential for hydrocarbon-consuming microorganisms; and

(c) an oxidation-reduction dye capable of indicating by a change in color a change in the redox potential caused by growth of a hydrocarbon-consuming microorganism in the tube.

15. A kit according to claim 14 wherein the selective growth substrate comprises an alcohol or an aldehyde.

16. A kit according to claim 14 wherein the dye comprises resazurin, methylene blue, or a tetrazolium dye

17. A kit according to claim 15 wherein the dye comprises resazurin, methylene blue, or a tetrazolium dye.