Laboratory Culture and Maintenance of Stenotrophomonas maltophilia

Osama Mahdi1, Bridget Eklund1, Nathan Fisher1

1 Department of Veterinary and Microbiological Sciences, North Dakota State University, Fargo, North Dakota
Publication Name:  Current Protocols in Microbiology
Unit Number:  Unit 6F.1
DOI:  10.1002/9780471729259.mc06f01s32
Online Posting Date:  February, 2014
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Abstract

Stenotrophomonas maltophilia is a ubiquitous soil bacterium that is increasingly recognized as an emerging nosocomial pathogen. This unit includes protocols for the in vitro growth and maintenance of S. maltophilia. Curr. Protoc. Microbiol. 32:6F.1.1‐6F.1.6. © 2014 by John Wiley & Sons, Inc.

Keywords: Stenotrophomonas maltophilia; laboratory growth; nosocomial; hospital acquired infection

     
 
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Table of Contents

  • Introduction
  • Strategic Planning
  • Basic Protocol 1: Growth of S. maltophilia from a Frozen Stock
  • Basic Protocol 2: Growth of S. maltophilia in Liquid Medium
  • Basic Protocol 3: Preparation of S. maltophilia Frozen Stocks
  • Commentary
  • Literature Cited
  • Tables
     
 
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Materials

Basic Protocol 1: Growth of S. maltophilia from a Frozen Stock

  Materials
  • S. maltophilia frozen stock (see protocol 3)
  • LB (Luria) agar plates ( appendix 4A)
  • Inoculating loop or needle, sterile
  • 37°C incubator

Basic Protocol 2: Growth of S. maltophilia in Liquid Medium

  Materials
  • LB (Lennox or Luria), nutrient, or trypticase soy broth, sterile (see appendix 4A)
  • S. maltophilia, grown on LB (Lennox or Luria) or nutrient agar, containing antibiotics if appropriate (see protocol 1)
  • Antibiotics, if required (see Table 6.1.2)
  • Capped test tubes or flask, sterile
  • Inoculating loop or needle, sterile
  • Vortex mixer
  • 37°C shaking incubator

Basic Protocol 3: Preparation of S. maltophilia Frozen Stocks

  Materials
  • S. maltophilia, grown on LB (Lennox or Luria) or nutrient agar, containing antibiotics if appropriate (see protocol 1)
  • LB (Lennox or Luria) broth (see appendix 4A)
  • 50% (v/v) glycerol, sterile
  • 15‐ml screw‐cap conical tubes
  • Inoculating loop or needle, sterile
  • 37°C shaking incubator
  • 1.5‐ml freezer vials, sterile
  • Vortex mixer
  • −80°C freezer
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Figures

Videos

Literature Cited

Literature Cited
  Allen, M.J., Tait, K., Muhling, M., Weynberg, K., Bradley, C., Trivedi, U., Gharbi, K., Nissimov, J., Mavromatis, K., Jensen, C.N., Grogan, G., and Ali, S.T. 2012. Genome sequence of Stenotrophomonas maltophilia PML168, which displays Baeyer‐Villiger monooxygenase activity. J. Bacteriol. 194:4753‐4754.
  Brooke, J.S. 2012. Stenotrophomonas maltophilia: An emerging global opportunistic pathogen. Clin. Microbiol. Rev. 25:2‐41.
  Chauhan, A., Green, S., Pathak, A., Thomas, J., and Venkatramanan, R. 2013. Whole‐genome sequence of five oyster‐associated bacteria show potential for crude oil hydrocarbon degradation. Genome Announc. 1:e00802‐13.
  Corsaro, D., Muller, K.D., and Michel, R. 2013. Molecular characterization and ultrastructure of a new amoeba endoparasite belonging to the Stenotrophomonas maltophilia complex. Exp. Parasitol. 133:389‐390.
  Crossman, L.C., Gould, V.C., Dow, J.M., Vernikos, G.S., Okazaki, A., Sebaihia, M., Saunders, D., Arrowsmith, C., Carver, T., Peters, N., Adlem, E., Kerhornou, A., Lord, A., Murphy, L., Seeger, K., Squares, R., Rutter, S., Quail, M.A., Rajandream, M., Harris, D., Churcher, C., Bentley, S.D., Parkhill, J., Thomson, N.R., and Avison, M.B. 2008. The complete genome, comparative and functional analysis of Stenotrophomonas maltophilia reveals an organism heavily shielded by drug resistance determinants. Genome Biol. 9:R74. doi:10.1186/gb‐2008‐9‐4‐r74.
  Lira, F., Hernandez, A., Belda, E., Sanches, M.B., Moya, A., Silva, F.J., and Martinez, J.L. 2012. Whole genome sequence of Stenotrophomonas maltophilia D457, a clinical isolate and a model strain. J. Bacteriol. 194:3563‐3564.
  Pompilio, A., Pomponio, S., Crocetta, V., Gherardi, G., Verginelli, F., Fiscarelli, E., Dicuonzo, G., Savini, V., D'Antonio, D., and Di Bonaventura, G. 2011. Phenotypic and genotypic characterization of Stenotrophomonas maltophilia isolates from patients with cystic fibrosis: Genome diversity, biofilm formation, and virulence. BMC Microbiol. 11:159.
  Rouf, R., Karaba, S.M., Dao, J., and Cianciotto, N.P. 2011. Stenotrophomonas maltophilia strains replicate and persist in the murine lung, but to significantly different degrees. Microbiology 157:2133‐2142.
  Sasserra, D., Leardini, I., Villa, L., Comandatore, F., Carta, C., Almeida, A., do Ceu Sousa, M., Gaiarsa, S., Marone, P., Pozio, E., and Caccio, S.M. 2013. Draft genome sequence of Stenotrophomonas maltophilia strain EPM1, found in association with a culture of the human parasite Giardia duodenalis. Genome Announc. 1:e00182‐13.
  Song, S., Yuan, X., Liu, S., Zhang, N., Wang, Y., Ke, Y., Xu, J., Huang, L., Chen, Z., and Li, Y. 2012. Genome sequence of Stenotrophomonas maltophilia S028, an isolate harboring the AmpR‐L2 resistance module. J. Bacteriol. 194:6696.
  Taghavi, S., Garafola, C., Monchy, S., Newman, L., Hoffman, A., Weyens, N., Barac, T., Vangrosveld, J., and van der Lelie, D. 2009. Genome survey and characterization of endophytic bacteria exhibiting a beneficial effect on growth and development of poplar trees. Appl. Environ. Microbiol. 75:748‐757.
  Zhang, L., Morrison, M., O Cuiv, P., Evans, P., and Rickard, C.M. 2013. Genome sequence of Stenotrophomonas maltophilia strain AU12‐09, isolated from an intravascular catheter. Genome Announc. 1:e00195‐13.
  Zhu, B., Liu, H., Tian, W.X., Fan, X.Y., Li, B., Zhou, X.P., Jin, G.L., and Xie, G.L. 2012. Genome sequence of Stenotrophomonas RR‐10, isolated as an endophyte from rice root. J. Bacteriol. 194:1280‐1281.
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