This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Davis, K. E. R.
Right arrow Articles by Janssen, P. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Davis, K. E. R.
Right arrow Articles by Janssen, P. H.
Agricola
Right arrow Articles by Davis, K. E. R.
Right arrow Articles by Janssen, P. H.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, February 2005, p. 826-834, Vol. 71, No. 2
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.2.826-834.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Effects of Growth Medium, Inoculum Size, and Incubation Time on Culturability and Isolation of Soil Bacteria

Kathryn E. R. Davis, Shayne J. Joseph, and Peter H. Janssen*

Department of Microbiology and Immunology, University of Melbourne, Parkville, Victoria, Australia

Received 9 July 2004/ Accepted 7 September 2004

Soils are inhabited by many bacteria from phylogenetic groups that are poorly studied because representatives are rarely isolated in cultivation studies. Part of the reason for the failure to cultivate these bacteria is the low frequency with which bacterial cells in soil form visible colonies when inoculated onto standard microbiological media, resulting in low viable counts. We investigated the effects of three factors on viable counts, assessed as numbers of CFU on solid media, and on the phylogenetic groups to which the isolated colony-forming bacteria belong. These factors were inoculum size, growth medium, and incubation time. Decreasing the inoculum size resulted in significant increases in the viable count but did not appear to affect colony formation by members of rarely isolated groups. Some media that are traditionally used for soil microbiological studies returned low viable counts and did not result in the isolation of members of rarely isolated groups. Newly developed media, in contrast, resulted in high viable counts and in the isolation of many members of rarely isolated groups, regardless of the inoculum size. Increased incubation times of up to 3 months allowed the development of visible colonies of members of rarely isolated groups in conjunction with the use of appropriate media. Once isolated, pure cultures of members of rarely isolated groups took longer to form visible colonies than did members of commonly isolated groups. Using these new media and extended incubation times, we were able to isolate many members of the phyla Acidobacteria (subdivisions 1, 2, 3, and 4), Gemmatimonadetes, Chloroflexi, and Planctomycetes (including representatives of the previously uncultured WPS-1 lineage) as well as members of the subclasses Rubrobacteridae and Acidimicrobidae of the phylum Actinobacteria.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of Melbourne, Parkville, Victoria 3010, Australia. Phone: 61 (3) 8344 5706. Fax: 61 (3) 9347 1540. E-mail: pjanssen{at}unimelb.edu.au.

Applied and Environmental Microbiology, February 2005, p. 826-834, Vol. 71, No. 2
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.2.826-834.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Zhang, J.-Y., Liu, X.-Y., Liu, S.-J. (2009). Nocardioides terrae sp. nov., isolated from forest soil. Int. J. Syst. Evol. Microbiol. 59: 2444-2448 [Abstract] [Full Text]  
  • Aoi, Y., Kinoshita, T., Hata, T., Ohta, H., Obokata, H., Tsuneda, S. (2009). Hollow-Fiber Membrane Chamber as a Device for In Situ Environmental Cultivation. Appl. Environ. Microbiol. 75: 3826-3833 [Abstract] [Full Text]  
  • Ward, N. L., Challacombe, J. F., Janssen, P. H., Henrissat, B., Coutinho, P. M., Wu, M., Xie, G., Haft, D. H., Sait, M., Badger, J., Barabote, R. D., Bradley, B., Brettin, T. S., Brinkac, L. M., Bruce, D., Creasy, T., Daugherty, S. C., Davidsen, T. M., DeBoy, R. T., Detter, J. C., Dodson, R. J., Durkin, A. S., Ganapathy, A., Gwinn-Giglio, M., Han, C. S., Khouri, H., Kiss, H., Kothari, S. P., Madupu, R., Nelson, K. E., Nelson, W. C., Paulsen, I., Penn, K., Ren, Q., Rosovitz, M. J., Selengut, J. D., Shrivastava, S., Sullivan, S. A., Tapia, R., Thompson, L. S., Watkins, K. L., Yang, Q., Yu, C., Zafar, N., Zhou, L., Kuske, C. R. (2009). Three Genomes from the Phylum Acidobacteria Provide Insight into the Lifestyles of These Microorganisms in Soils. Appl. Environ. Microbiol. 75: 2046-2056 [Abstract] [Full Text]  
  • Nichols, D., Lewis, K., Orjala, J., Mo, S., Ortenberg, R., O'Connor, P., Zhao, C., Vouros, P., Kaeberlein, T., Epstein, S. S. (2008). Short Peptide Induces an "Uncultivable" Microorganism To Grow In Vitro. Appl. Environ. Microbiol. 74: 4889-4897 [Abstract] [Full Text]  
  • Bollmann, A., Lewis, K., Epstein, S. S. (2007). Incubation of Environmental Samples in a Diffusion Chamber Increases the Diversity of Recovered Isolates. Appl. Environ. Microbiol. 73: 6386-6390 [Abstract] [Full Text]  
  • Eichorst, S. A., Breznak, J. A., Schmidt, T. M. (2007). Isolation and Characterization of Soil Bacteria That Define Terriglobus gen. nov., in the Phylum Acidobacteria. Appl. Environ. Microbiol. 73: 2708-2717 [Abstract] [Full Text]  
  • Zwolinski, M. D. (2007). DNA Sequencing: Strategies for Soil Microbiology. Soil Sci. 71: 592-600 [Abstract] [Full Text]  
  • Drees, K. P., Neilson, J. W., Betancourt, J. L., Quade, J., Henderson, D. A., Pryor, B. M., Maier, R. M. (2006). Bacterial Community Structure in the Hyperarid Core of the Atacama Desert, Chile. Appl. Environ. Microbiol. 72: 7902-7908 [Abstract] [Full Text]  
  • Buckley, D. H., Huangyutitham, V., Nelson, T. A., Rumberger, A., Thies, J. E. (2006). Diversity of planctomycetes in soil in relation to soil history and environmental heterogeneity.. Appl. Environ. Microbiol. 72: 4522-4531 [Abstract] [Full Text]  
  • Shimomura, Y., Ohno, R., Kawai, F., Kimbara, K. (2006). Method for assessment of viability and morphological changes of bacteria in the early stage of colony formation on a simulated natural environment.. Appl. Environ. Microbiol. 72: 5037-5042 [Abstract] [Full Text]  
  • Sanguin, H., Remenant, B., Dechesne, A., Thioulouse, J., Vogel, T. M., Nesme, X., Moenne-Loccoz, Y., Grundmann, G. L. (2006). Potential of a 16S rRNA-Based Taxonomic Microarray for Analyzing the Rhizosphere Effects of Maize on Agrobacterium spp. and Bacterial Communities.. Appl. Environ. Microbiol. 72: 4302-4312 [Abstract] [Full Text]  
  • Cavaletti, L., Monciardini, P., Bamonte, R., Schumann, P., Rohde, M., Sosio, M., Donadio, S. (2006). New lineage of filamentous, spore-forming, gram-positive bacteria from soil.. Appl. Environ. Microbiol. 72: 4360-4369 [Abstract] [Full Text]  
  • Janssen, P. H. (2006). Identifying the Dominant Soil Bacterial Taxa in Libraries of 16S rRNA and 16S rRNA Genes.. Appl. Environ. Microbiol. 72: 1719-1728 [Full Text]  
  • Sait, M., Davis, K. E. R., Janssen, P. H. (2006). Effect of pH on Isolation and Distribution of Members of Subdivision 1 of the Phylum Acidobacteria Occurring in Soil.. Appl. Environ. Microbiol. 72: 1852-1857 [Abstract] [Full Text]  
  • Kopke, B., Wilms, R., Engelen, B., Cypionka, H., Sass, H. (2005). Microbial Diversity in Coastal Subsurface Sediments: a Cultivation Approach Using Various Electron Acceptors and Substrate Gradients. Appl. Environ. Microbiol. 71: 7819-7830 [Abstract] [Full Text]  
  • Sangwan, P., Kovac, S., Davis, K. E. R., Sait, M., Janssen, P. H. (2005). Detection and Cultivation of Soil Verrucomicrobia. Appl. Environ. Microbiol. 71: 8402-8410 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»