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Use of Blue Agar CAS Assay for Siderophore Detection

    Authors: Brian C. Louden1, Daniel Haarmann1, Aaron M. Lynne1,*
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    Affiliations: 1: Department of Biological Sciences, Sam Houston State University, Huntsville, TX 77341
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • Published 19 May 2011
    • *Corresponding author. Mailing address: Box 2116, Department of Biological Sciences, Sam Houston State University, 1806 Avenue J, Huntsville, TX 77341. Phone: 936-294-1544. Fax: 936-294-3940. E-mail: aml027@shsu.edu.
    • Copyright © 2011, American Society for Microbiology
    Source: J. Microbiol. Biol. Educ. May 2011 vol. 12 no. 1 51-53. doi:10.1128/jmbe.v12i1.249
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    Abstract:

    Schwyn and Neiland developed a universal siderophore assay using chrome azurol S(CAS) and hexadecyltrimethylammonium bromide (HDTMA) as indicators. In Schwyn and Neiland’s original paper, the procedure given for making CAS agar is written in general terms and can be difficult to follow, especially for an individual who has limited experience making more complex media. Here, we give a step-by-step protocol for making the CAS agar plates, and we discuss how this media can be incorporated in a comprehensive project in a microbiology lab course for biology majors.

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Escherichia coli
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References & Citations

1. Ames-Gottfred NP, Christie BR, Jordan DC 1989 Use of the Chrome Azurol S agar plate technique to differentiate strains and field isolates of Rhizobium leguminosarum biovar trifolii Appl Environ Microbiol 55 707 710 16347877
2. Arnold LD, Viswanatha T 1983 The use of bis(mercaptoacetato-S,O)hydroxoiron(III) complex for the determination of hydroxamates J Biochem Biophys Methods 8 307 320 10.1016/0165-022X(83)90005-2 6663004 http://dx.doi.org/10.1016/0165-022X(83)90005-2
3. Arnow LE 1937 Colorimetric determination of the components of 3,4-Dihydroxyphenylalaninetyrosine mixtures J Biol Chem 118 531 537
4. Atkin CL, Neilands JB, Phaff HJ 1970 Rhodotorulic acid from species of Leucosporidium, Rhodosporidium, Rhodotorula, Sporidiobolus, and Sporobolomyces, and a new alanine-containing ferrichrome from Cryptococcus melibiosum J Bacteriol 103 722 733 5529038
5. Machuca A, Milagres AM 2003 Use of CAS-agar plate modified to study the effect of different variables on the siderophore production by Aspergillus Letters Appl Microbiol 36 177 181 10.1046/j.1472-765X.2003.01290.x http://dx.doi.org/10.1046/j.1472-765X.2003.01290.x
6. Miethke M, Marahiel MA 2007 Siderophore-based iron acquisition and pathogen control Microbiol Molecul Biol Rev 71 413 451 10.1128/MMBR.00012-07 http://dx.doi.org/10.1128/MMBR.00012-07
7. Pérez-Miranda S, Cabirol N, George-Téllez R, Zamudio-Rivera LS, Fernández FJ 2007 O-CAS, a fast and universal method for siderophore detection J Microbiol Methods 70 127 131 10.1016/j.mimet.2007.03.023 17507108 http://dx.doi.org/10.1016/j.mimet.2007.03.023
8. Schwyn B, Neilands JB 1987 Universal chemical assay for the detection and determination of siderophores Analytical Biochem 160 47 56 10.1016/0003-2697(87)90612-9 http://dx.doi.org/10.1016/0003-2697(87)90612-9
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/content/journal/jmbe/10.1128/jmbe.v12i1.249
2011-05-19
2017-09-25

Abstract:

Schwyn and Neiland developed a universal siderophore assay using chrome azurol S(CAS) and hexadecyltrimethylammonium bromide (HDTMA) as indicators. In Schwyn and Neiland’s original paper, the procedure given for making CAS agar is written in general terms and can be difficult to follow, especially for an individual who has limited experience making more complex media. Here, we give a step-by-step protocol for making the CAS agar plates, and we discuss how this media can be incorporated in a comprehensive project in a microbiology lab course for biology majors.

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FIGURE 1

Example CAS plate with four different bacteria plated. Moving clockwise from top left: .

Source: J. Microbiol. Biol. Educ. May 2011 vol. 12 no. 1 51-53. doi:10.1128/jmbe.v12i1.249
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