Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1992 Dec;58(12):4011–4015. doi: 10.1128/aem.58.12.4011-4015.1992

Evaluation of Culture Techniques for Isolation of Pseudomonas pseudomallei from Soil

Leslie R Ashdown 1,*, Sue G Clarke 1
PMCID: PMC183218  PMID: 16348827

Abstract

Three selective enrichment broths and four selective agar media were evaluated for their ability to support the growth of Pseudomonas pseudomallei both at 35°C and at ambient temperature (range, 20 to 32°C; mean, 25°C). Colony counts of 50 strains of P. pseudomallei and recovery studies with 1 soil strain in 60 simulated soil samples demonstrated that enrichment with Trypticase soy broth incorporating 5 mg of crystal violet per liter and 20 mg of colistin per liter (CVCB) and subculture to Ashdown medium supported the growth of all 50 strains and produced the highest recovery rates with the greatest suppression of other soil flora. An enrichment broth of MacConkey broth (purple) incorporating 10 mg of crystal violet per liter, 5 mg of bromcresol purple per liter, 25 mg of gentamicin per liter, and 650 mg of streptomycin per liter showed greater suppression of soil bacteria than CVCB, but it failed to support the growth of three strains of P. pseudomallei. Recovery rates were essentially the same irrespective of whether the soil samples were incubated at 35°C or at ambient temperature, provided cultures were incubated in protected shade for an extended period. This is an important feature for field work in large-scale epidemiological surveys in which resources are limited.

Full text

PDF
4011

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ashdown L. R. An improved screening technique for isolation of Pseudomonas pseudomallei from clinical specimens. Pathology. 1979 Apr;11(2):293–297. doi: 10.3109/00313027909061954. [DOI] [PubMed] [Google Scholar]
  2. Ashdown L. R. Evaluation of antibiotics in Pseudomonas infections. Med J Aust. 1979 Apr 7;1(7):284–284. doi: 10.5694/j.1326-5377.1979.tb112096.x. [DOI] [PubMed] [Google Scholar]
  3. Ashdown L. R., Guard R. W. The prevalence of human melioidosis in Northern Queensland. Am J Trop Med Hyg. 1984 May;33(3):474–478. doi: 10.4269/ajtmh.1984.33.474. [DOI] [PubMed] [Google Scholar]
  4. Ashdown L. R. Identification of Pseudomonas pseudomallei in the clinical laboratory. J Clin Pathol. 1979 May;32(5):500–504. doi: 10.1136/jcp.32.5.500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ashdown L. R. Nosocomial infection due to Pseudomonas pseudomallei: two cases and an epidemiologic study. Rev Infect Dis. 1979 Sep-Oct;1(5):891–894. doi: 10.1093/clinids/1.5.891. [DOI] [PubMed] [Google Scholar]
  6. Ashdown L. R. Rapid differentiation of Pseudomonas pseudomallei from Pseudomonas cepacia. Lett Appl Microbiol. 1992 May;14(5):203–205. doi: 10.1111/j.1472-765x.1992.tb00685.x. [DOI] [PubMed] [Google Scholar]
  7. Ellison D. W., Baker H. J., Mariappan M. Melioidosis in Malaysia. I. A method for isolation of Pseudomonas pseudomallei from soil and surface water. Am J Trop Med Hyg. 1969 Sep;18(5):694–697. [PubMed] [Google Scholar]
  8. Guard R. W., Khafagi F. A., Brigden M. C., Ashdown L. R. Melioidosis in Far North Queensland. A clinical and epidemiological review of twenty cases. Am J Trop Med Hyg. 1984 May;33(3):467–473. doi: 10.4269/ajtmh.1984.33.467. [DOI] [PubMed] [Google Scholar]
  9. Leelarasamee A., Bovornkitti S. Melioidosis: review and update. Rev Infect Dis. 1989 May-Jun;11(3):413–425. doi: 10.1093/clinids/11.3.413. [DOI] [PubMed] [Google Scholar]
  10. McCormick J. B., Weaver R. E., Hayes P. S., Boyce J. M., Feldman R. A. Wound infection by an indigenous Pseudomonas pseudomallei-like organism isolated from the soil: case report and epidemiologic study. J Infect Dis. 1977 Jan;135(1):103–107. doi: 10.1093/infdis/135.1.103. [DOI] [PubMed] [Google Scholar]
  11. Thomas A. D., Forbes-Faulkner J. C. Persistence of Pseudomonas pseudomallei in soil. Aust Vet J. 1981 Nov;57(11):535–536. doi: 10.1111/j.1751-0813.1981.tb05804.x. [DOI] [PubMed] [Google Scholar]
  12. Thomas A. D., Forbes-Faulkner J., Parker M. Isolation of Pseudomonas pseudomallei from clay layers at defined depths. Am J Epidemiol. 1979 Oct;110(4):515–521. doi: 10.1093/oxfordjournals.aje.a112832. [DOI] [PubMed] [Google Scholar]
  13. Wuthiekanun V., Dance D. A., Wattanagoon Y., Supputtamongkol Y., Chaowagul W., White N. J. The use of selective media for the isolation of Pseudomonas pseudomallei in clinical practice. J Med Microbiol. 1990 Oct;33(2):121–126. doi: 10.1099/00222615-33-2-121. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES