Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1997 Mar;63(3):851–856. doi: 10.1128/aem.63.3.851-856.1997

Investigation of persistent colonization by Pseudomonas aeruginosa-like strains in a spring water bottling plant.

P V Morais 1, C Mesquita 1, J L Andrade 1, M S da Costa 1
PMCID: PMC168380  PMID: 9055406

Abstract

Ninety-seven strains, producing a fluorescent pigment under UV light and/or a green diffusive pigment on cetrimide-naladixic acid agar, were isolated from a spring water bottling plant. These strains were presumptively identified as Pseudomonas aeruginosa, but they could not be confirmed as strains of this species nor identified by the API 20NE identification system. The isolates and reference strains were clustered by computer-assisted whole-cell protein sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The numerical analysis of the protein electrophoregrams resulted in the formation of four clusters at a similarity level of 80% and two unclustered type strains. One cluster included strains isolated during a 4-month period and reference strains of several biotypes of P. fluorescens. The remaining isolates formed another cluster with a very high similarity of level, which included two groups of strains based on biochemical characterization by the API 20NE Test System. Strains were typed by random amplified polymorphic DNA (RAPD)-PCR and two different RAPD patterns were obtained, corresponding to each biochemical profile. This persistent colonization seems to be caused by a single species present in the bottling system, with two clonal origins, not related to P. aeruginosa or to any of the other type strains tested. Partial 16S rDNA sequence of a representative strain of one cluster of isolates had a level of similarity of 99.3% with P. alcaligenes. This study shows that characteristics similar to P. aeruginosa on cetrimide-naladixic acid agar can be exhibited by several groups of fluorescent pseudomonads that do not belong to this species, clearly showing that confirmation tests must be performed before a decision regarding the water quality is made.

Full Text

The Full Text of this article is available as a PDF (357.1 KB).

Selected References

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

  1. Bischofberger T., Cha S. K., Schmitt R., König B., Schmidt-Lorenz W. The bacterial flora of non-carbonated, natural mineral water from the springs to reservoir and glass and plastic bottles. Int J Food Microbiol. 1990 Aug;11(1):51–71. doi: 10.1016/0168-1605(90)90039-8. [DOI] [PubMed] [Google Scholar]
  2. Christensen H., Boye M., Poulsen L. K., Rasmussen O. F. Analysis of fluorescent pseudomonads based on 23S ribosomal DNA sequences. Appl Environ Microbiol. 1994 Jun;60(6):2196–2199. doi: 10.1128/aem.60.6.2196-2199.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Costas M., Holmes B., Frith K. A., Riddle C., Hawkey P. M. Identification and typing of Proteus penneri and Proteus vulgaris biogroups 2 and 3, from clinical sources, by computerized analysis of electrophoretic protein patterns. J Appl Bacteriol. 1993 Nov;75(5):489–498. doi: 10.1111/j.1365-2672.1993.tb02806.x. [DOI] [PubMed] [Google Scholar]
  4. Elaichouni A., Verschraegen G., Claeys G., Devleeschouwer M., Godard C., Vaneechoutte M. Pseudomonas aeruginosa serotype O12 outbreak studied by arbitrary primer PCR. J Clin Microbiol. 1994 Mar;32(3):666–671. doi: 10.1128/jcm.32.3.666-671.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Elomari M., Coroler L., Izard D., Leclerc H. A numerical taxonomic study of fluorescent Pseudomonas strains isolated from natural mineral waters. J Appl Bacteriol. 1995 Jan;78(1):71–81. doi: 10.1111/j.1365-2672.1995.tb01676.x. [DOI] [PubMed] [Google Scholar]
  6. Ferreira A. C., Morais P. V., Gomes C., da Costa M. S. Computer-aided comparison of protein electrophoretic patterns for grouping and identification of heterotrophic bacteria from mineral water. J Appl Bacteriol. 1996 May;80(5):479–486. doi: 10.1111/j.1365-2672.1996.tb03246.x. [DOI] [PubMed] [Google Scholar]
  7. González C., Gutiérrez C., Grande T. Bacterial flora in bottled uncarbonated mineral drinking water. Can J Microbiol. 1987 Dec;33(12):1120–1125. doi: 10.1139/m87-196. [DOI] [PubMed] [Google Scholar]
  8. Havelaar A. H., During M., Delfgou-Van Asch E. H. Comparative study of membrane filtration and enrichment media for the isolation and enumeration of Pseudomonas aeruginosa from sewage, surface water, and swimming pools. Can J Microbiol. 1985 Aug;31(8):686–692. doi: 10.1139/m85-130. [DOI] [PubMed] [Google Scholar]
  9. Hunter P. R. The microbiology of bottled natural mineral waters. J Appl Bacteriol. 1993 Apr;74(4):345–352. doi: 10.1111/j.1365-2672.1993.tb05137.x. [DOI] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Manaia C. M., Nunes O. C., Morais P. V., da Costa M. S. Heterotrophic plate counts and the isolation of bacteria from mineral waters on selective and enrichment media. J Appl Bacteriol. 1990 Dec;69(6):871–876. doi: 10.1111/j.1365-2672.1990.tb01586.x. [DOI] [PubMed] [Google Scholar]
  12. Morais P. V., da Costa M. S. Alterations in the major heterotrophic bacterial populations isolated from a still bottled mineral water. J Appl Bacteriol. 1990 Nov;69(5):750–757. doi: 10.1111/j.1365-2672.1990.tb01572.x. [DOI] [PubMed] [Google Scholar]
  13. Moreira L., Agostinho P., Morais P. V., da Costa M. S. Survival of allochthonous bacteria in still mineral water bottled in polyvinyl chloride (PVC) and glass. J Appl Bacteriol. 1994 Sep;77(3):334–339. doi: 10.1111/j.1365-2672.1994.tb03082.x. [DOI] [PubMed] [Google Scholar]
  14. Römling U., Wingender J., Müller H., Tümmler B. A major Pseudomonas aeruginosa clone common to patients and aquatic habitats. Appl Environ Microbiol. 1994 Jun;60(6):1734–1738. doi: 10.1128/aem.60.6.1734-1738.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Veríssimo A., Morais P. V., Diogo A., Gomes C., da Costa M. S. Characterization of Legionella species by numerical analysis of whole-cell protein electrophoresis. Int J Syst Bacteriol. 1996 Jan;46(1):41–49. doi: 10.1099/00207713-46-1-41. [DOI] [PubMed] [Google Scholar]
  16. Warburton D. W. A review of the microbiological quality of bottled water sold in Canada. Part 2. The need for more stringent standards and regulations. Can J Microbiol. 1993 Feb;39(2):158–168. doi: 10.1139/m93-022. [DOI] [PubMed] [Google Scholar]
  17. Wiedmann-al-Ahmad M., Tichy H. V., Schön G. Characterization of Acinetobacter type strains and isolates obtained from wastewater treatment plants by PCR fingerprinting. Appl Environ Microbiol. 1994 Nov;60(11):4066–4071. doi: 10.1128/aem.60.11.4066-4071.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Williams J. G., Kubelik A. R., Livak K. J., Rafalski J. A., Tingey S. V. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 1990 Nov 25;18(22):6531–6535. doi: 10.1093/nar/18.22.6531. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES