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PHYSIOLOGY AND BIOTECHNOLOGY

N-Acetyl-l-Cysteine Affects Growth, Extracellular Polysaccharide Production, and Bacterial Biofilm Formation on Solid Surfaces

Ann-Cathrin Olofsson, Malte Hermansson, Hans Elwing
Ann-Cathrin Olofsson
1Department of Cell and Molecular Biology—Interface Biophysics
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  • For correspondence: anki.olofsson@gmm.gu.se
Malte Hermansson
2Department of Cell and Molecular Biology—Microbiology, Göteborg University, 405 30 Göteborg, Sweden
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Hans Elwing
1Department of Cell and Molecular Biology—Interface Biophysics
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DOI: 10.1128/AEM.69.8.4814-4822.2003
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    FIG. 1.

    Effect of NAC on bacteria cultured in 20% LB. Bacterial growth, expressed as optical density at 600 nm [OD(600nm)], was measured for 24 h. Symbols: •, no NAC;, ×, 0.25 mg of NAC ml−1; □, 0.5 mg of NAC ml−1; ▪, 1.0 mg of NAC ml−1; ○, 2.0 mg of NAC ml−1. The growth of A. lwoffii (a), A. baumannii (b), E. cloacae (c), K. pneumoniae (d), P. mendocina (e), a multispecies community consisting of A. baumannii, P. mendocina, K. pneumoniae, A. lwoffii, E. cloacae,S. warneri, and Bacillus sp. (f), Bacillus sp. (g), B. cereus (h), B. megaterium (i), B. subtilis (j), and S. warneri (k) was examined. The error bars indicate the standard deviations of the means for three experiments.

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    FIG. 2.

    Effect of NAC on EPS production. Bacteria were grown in 20% LB overnight in the presence or absence of NAC and then starved in PS for 48 h. Open columns, no NAC; cross-hatched columns, 0.25 mg of NAC ml−1; grey columns, 0.5 mg of NAC ml−1. The bars indicate the standard deviations of the means for four experiments. Abbreviations: K.p, K. pneumoniae; A.b, A. baumannii; B.m, B. megaterium; B.s, B. subtilis; B.c, B. cereus; A.l, A. lwoffii; P.m, P. mendocina; E.c, E. cloacae; S.w, S. warneri; B, Bacillus sp. NM, not measured.

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    FIG. 3.

    Effect of NAC on EPS production by starving bacteria. Bacteria were grown in 20% LB overnight in the absence of NAC, washed, and subsequently exposed to 0.5 mg of NAC ml−1 in PS for 48 h. Open columns, no NAC; cross-hatched columns, 0.5 mg of NAC ml−1. The bars indicate the standard deviations of the means for three experiments. Abbreviations: B.c, B. cereus; A.l, A. lwoffii; P.m, P. mendocina; S.w, S. warneri; B, Bacillus sp.

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    FIG. 4.

    Effect of NAC on degradation of EPS. Isolated EPS from K. pneumoniae, B. megaterium, B. subtilis, B. cereus, E. cloacae, and P. mendocina, as well as dextran, were incubated in the presence or absence of 0.5 mg of NAC ml−1 for 1 h at room temperature. Open columns, no NAC; cross-hatched columns, 0.5 mg of NAC ml−1. The bars indicate the standard deviations of the means for three experiments. The values for dextran (0.5 mg ml−1) were not affected by NAC (the values were too small to show). Abbreviations: K.p, K. pneumoniae; B.m, B. megaterium; B.s, B. subtilis; B.c, B. cereus; E.c, E. cloacae; P.m, P. mendocina.

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    FIG. 5.

    Numbers of cells that adhered to stainless steel surfaces after 24, 48, and 72 h of incubation in 20% LB. The seven-member multispecies community comprised A. baumannii, A. lwoffii, Bacillus sp., E. cloacae, K. pneumoniae, P. mendocina, and S. warneri. Open columns, no NAC; cross-hatched columns, 0.5 mg of NAC ml−1. The bars indicate the standard deviations of the means for three experiments. Abbreviations: K.p, K. pneumoniae; E.c, E. cloacae; mix, multispecies community.

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    FIG. 6.

    Numbers of cells that adhered to stainless steel surfaces after 24, 48, and 72 h of incubation in process water. The seven-member multispecies community comprised A. baumannii, A. lwoffii, Bacillus sp., E. cloacae, K. pneumoniae, P. mendocina, and S. warneri. Open columns, no NAC; cross-hatched columns, 0.5 mg of NAC ml−1. The bars indicate the standard deviations of the means for three experiments.

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    FIG. 7.

    Acridine orange-stained K. pneumoniae attached to stainless steel surfaces after 72 h of incubation in 20% LB. (a) No NAC in the medium; (b) 0.5 mg of NAC ml−1 in the medium.

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    FIG. 8.

    Attachment of K. pneumoniae to stainless steel surfaces after 72 h of incubation in 20% LB in the absence of NAC (a) and in the presence of 0.5 mg of NAC ml−1 (b). WGA lectin labeled with the fluorescent probe tetramethyl rhodamine isocyanate was used to visualize CPS and EPS.

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    Bacterial strains used in this study

    TaxonIsolation sourceCCUG ref- erence no.Reference or source
    Gram-negative bacteria
        Acinetobacter baumanniiBiofilm in white water39226This study
        Acinetobacter lwoffiiBiofilm in wire water39223This study
        Enterobacter cloacaeBiofilm in wire water39230This study
        Klebsiella pneumoniaeBiofilm in white water39227This study
        Pseudomonas mendocinaBiofilm in white water39225This study
    Gram-positive bacteria
        Bacillus sp.Biofilm in white water39224This study
        Bacillus cereus6514CCUG
        Bacillus megaterium1817CCUG
        Bacillus subtilis1638CCUG
        Staphylococcus warneriBiofilm in white water39229This study
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N-Acetyl-l-Cysteine Affects Growth, Extracellular Polysaccharide Production, and Bacterial Biofilm Formation on Solid Surfaces
Ann-Cathrin Olofsson, Malte Hermansson, Hans Elwing
Applied and Environmental Microbiology Aug 2003, 69 (8) 4814-4822; DOI: 10.1128/AEM.69.8.4814-4822.2003

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N-Acetyl-l-Cysteine Affects Growth, Extracellular Polysaccharide Production, and Bacterial Biofilm Formation on Solid Surfaces
Ann-Cathrin Olofsson, Malte Hermansson, Hans Elwing
Applied and Environmental Microbiology Aug 2003, 69 (8) 4814-4822; DOI: 10.1128/AEM.69.8.4814-4822.2003
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KEYWORDS

Acetylcysteine
bacteria
biofilms
Polysaccharides, Bacterial

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