Laboratory Maintenance of Acinetobacter baumannii

Anna C. Jacobs1, Daniel V. Zurawski1

1 Walter Reed Army Institute of Research, Department of Wound Infections, Silver Spring, Maryland
Publication Name:  Current Protocols in Microbiology
Unit Number:  Unit 6G.1
DOI:  10.1002/9780471729259.mc06g01s35
Online Posting Date:  November, 2014
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Abstract

Acinetobacter baumannii has recently drawn great interest in the microbiology research community due to the increase in clinical antibiotic resistance of this organism, and persistence of this bacterial species in the hospital environment. This unit outlines protocols for the growth and maintenance of A. baumannii in the laboratory. © 2014 by John Wiley & Sons, Inc.

Keywords: Acinetobacter baumannii; laboratory growth; minimal medium; glycerol stocks; storage

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Growth of A. baumannii from a Frozen Stock
  • Alternate Protocol 1: Direct Growth of A. baumannii in Liquid Medium from a Frozen Stock
  • Basic Protocol 2: Growth of A. baumannii in Liquid Medium from a Single Colony
  • Basic Protocol 3: Making Frozen Stocks of A. baumannii
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Growth of A. baumannii from a Frozen Stock

  Materials
  • Frozen stock of A. baumannii (see protocol 4)
  • LB agar plate (see recipe)
  • Sterilized inoculation needle or wooden stick

Alternate Protocol 1: Direct Growth of A. baumannii in Liquid Medium from a Frozen Stock

  Materials
  • Frozen stock of A. baumannii (see protocol 4)
  • Lysogeny broth (LB; see recipe)
  • 14‐ml sterile plastic round‐bottom tubes
  • Sterile inoculation needle or wooden stick
  • Pipetman and serological pipets

Basic Protocol 2: Growth of A. baumannii in Liquid Medium from a Single Colony

  Materials
  • A. baumannii grown on LB agar plate (see protocol 1)
  • LB (see recipe)
  • Sterile inoculation needle or wooden stick
  • Pipetman and serological pipets
  • 14‐ml sterile plastic round‐bottom tubes

Basic Protocol 3: Making Frozen Stocks of A. baumannii

  Materials
  • A. baumannii grown on LB agar plate (see protocol 1)
  • LB (see recipe)
  • 80% glycerol, sterile (see recipe)
  • Sterile inoculation needle or wooden stick
  • Pipetman and serological pipets
  • 14‐ml sterile plastic round‐bottom tubes
  • 2‐ml freezer tube
  • Vortex
  • −80°C freezer
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Figures

Videos

Literature Cited

Literature Cited
  Barbaro, S.E., Trevors, J.T., and Inniss, W.E. 2002. Effect of different carbon sources and cold shock on protein synthesis by a psychrotrophic Acinetobacter sp. Can. J. Microbiol. 48:239‐244.
  Dijkshoorn, L., Van Ooyen, A., Hop, W.C., Theuns, M., and Michel, M.F. 1990. Comparison of clinical Acinetobacter strains using a carbon source growth assay. Epidemiol. Infect. 104:443‐453.
  Djeribi, R., Boucherit, Z., Bouchloukh, W., Zouaoui, W., Latrache, H., Hamadi, F., and Menaa, B. 2013. A study of pH effects on the bacterial surface physicochemical properties of Acinetobacter baumannii. Colloids Surf. B Biointerfaces 102:540‐545.
  Gerner‐Smidt, P., Tjernberg, I., and Ursing, J. 1991. Reliability of phenotypic tests for identification of Acinetobacter species. J. Clin. Microbiol. 29:277‐282.
  Hujer, K.M., Hujer, A.M., Hulten, E.A., Bajaksouzian, S., Adams, J.M., Donskey, C.J., Ecker, D.J., Massire, C., Eshoo, M.W., Sampath, R., Thomson, J.M., Rather, P.N., Craft, D.W., Fishbain, J.T., Ewell, A.J., Jacobs, M.R., Paterson, D.L., and Bonomo, R.A. 2006. Analysis of antibiotic resistance genes in multidrug‐resistant Acinetobacter sp. isolates from military and civilian patients treated at the Walter Reed Army Medical Center. Antimicrob. Agents Chemother. 50:4114‐4123.
  Keen, E.F., III, Murray, C.K., Robinson, B.J., Hospenthal, D.R., Co, E.M., and Aldous, W.K. 2010. Changes in the incidences of multidrug‐resistant and extensively drug‐resistant organisms isolated in a military medical center. Infect. Control Hosp. Epidemiol. 31:728‐732.
  McConnell, M.J., Actis, L., and Pachón, J. 2013. Acinetobacter baumannii: Human infections, factors contributing to pathogenesis and animal models. FEMS Microbiol. Rev. 37:130‐155.
  O'Shea, M.K. 2012. Acinetobacter in modern warfare. Int. J. Antimicrob. Agents 39:363‐375.
  Obeidat, N., Jawdat, F., Al‐Bakri, A.G., and Shehabi, A.A. 2014. Major biologic characteristics of Acinetobacter baumannii isolates from hospital environmental and patients' respiratory tract sources. Am. J. Infect. Control 42:401‐404.
  Seifert, H. and Dijkshoorn, L. 2008. Overview of the microbial characteristics, taxonomy, and epidemiology of Acinetobacter. In Acinetobacter Biology and Pathogenesis (M. Bendinelli, H. Friedman, and E. Bergogne‐Bérézin, eds.) pp. 19‐45. Springer, New York.
  Spellberg, B. and Rex, J.H. 2013. The value of single‐pathogen antibacterial agents. Nat. Rev. Drug Discov. 12:963.
  Rocha, L.L., Colares, G.B., Angelim, A.L., Grangeiro, T.B., and Melo, V.M. 2013. Culturable populations of Acinetobacter can promptly respond to contamination by alkanes in mangrove sediments. Mar. Pollut. Bull. 76:214‐219.
  Tomaras, A.P., Flagler, M.J., Dorsey, C.W., Gaddy, J.A., and Actis, L.A. 2008. Characterization of a two‐component regulatory system from Acinetobacter baumannii that controls biofilm formation and cellular morphology. Microbiology 154:3398‐3409.
  Yun, H.C., Branstetter, J.G., and Murray, C.K. 2008. Osteomyelitis in military personnel wounded in Iraq and Afghanistan. J. Trauma 64:S163‐S168.
  Zhang, J., Gorkovenko, A., Gross, R.A., Allen, A.L., and Kaplan, D. 1997. Incorporation of 2‐hydroxyl fatty acids by Acinetobacter calcoaceticus RAG‐1 to tailor emulsan structure. Int. J. Biol. Macromol. 20:9‐21.
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