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    Wilking Fundamentalintro To Biofilms 2016 v3

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    Gustavo
    The document discusses biofilms and their impact on industry, environment, and health. It describes how biofilms form and undergo life cycles of growth, spreading, and detachment. It also explains how biofilms are more tolerant than planktonic bacteria to antimicrobial agents due to their ability to slow penetration and alter microenvironments through stress responses.

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd

    Wilking Fundamentalintro To Biofilms 2016 v3

    Uploaded by

    Gustavo
    21 views48 pages
    The document discusses biofilms and their impact on industry, environment, and health. It describes how biofilms form and undergo life cycles of growth, spreading, and detachment. It also explains how biofilms are more tolerant than planktonic bacteria to antimicrobial agents due to their ability to slow penetration and alter microenvironments through stress responses.

    Original Description:

    Introducción: Fundamentos Biofilms

    Original Title

    Wilking Fundamentalintro to Biofilms 2016 v3

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    The document discusses biofilms and their impact on industry, environment, and health. It describes how biofilms form and undergo life cycles of growth, spreading, and detachment. It also explains how biofilms are more tolerant than planktonic bacteria to antimicrobial agents due to their ability to slow penetration and alter microenvironments through stress responses.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    21 views48 pages

    Wilking Fundamentalintro To Biofilms 2016 v3

    Uploaded by

    Gustavo
    The document discusses biofilms and their impact on industry, environment, and health. It describes how biofilms form and undergo life cycles of growth, spreading, and detachment. It also explains how biofilms are more tolerant than planktonic bacteria to antimicrobial agents due to their ability to slow penetration and alter microenvironments through stress responses.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 48

    Center for Biofilm Engineering

    Fundamentals of
    Microbial Biofilms
    Center for Biofilm Engineering at
    Montana State University

    James N. Wilking
    Assistant Professor, ChBE
    Center for Biofilm Engineering
    Montana State University
    wilkinglab.com
    2011, CLSM biofilm: K Hunt, MSU-CBE PhD student

    CFP Biennial Meeting | 2016

    Center for Biofilm Engineering


    Bozeman, Montana

    Free-swimming microbes vs. biofilms

    Biofilms are all around us

    Photos by CBE staff and students


    unless otherwise noted.

    Extracellular matrix:
    a hallmark of microbial biofilms

    JW Costerton

    Extracellular matrix:
    a hallmark of microbial biofilms

    Extracellular matrix:
    a hallmark of microbial biofilms

    The extracellular matrix provides the


    biofilm with mechanical integrity

    Biofilms undergo a life-cycle

    Biofilms undergo a life-cycle

    Biofilm growth and spreading

    Biofilms form complex structures

    Biofilms undergo a life-cycle

    Biofilm detachment

    Planktonic bacteria ready to be released!

    Biofilms impact industry, environment & health

    Center for Biofilm Engineering


    Industrial Associates
    Consumer Products
    Church & Dwight
    Colgate-Palmolive
    Johnson & Johnson
    Kimberly-Clark
    Masco
    Procter & Gamble
    Reckitt Benckiser
    Sherwin Williams
    Testing Laboratories
    WuXi AppTec*
    Energy
    ExxonMobil
    BP

    Specialty Chemicals
    BASF
    BCG Solutions *
    Clorox
    Dow Corning
    Dow Microbial Control
    Ecolab/Nalco
    NCH Corp.
    Novozymes A/S
    Sani-Marc
    Sample6 Technologies*
    Sealed-Air
    US Govt Programs/Labs
    NASA

    Health Care/Biomedical
    3M
    Bard Access Systems
    Bausch & Lomb
    Baxter Healthcare
    Bend Research*
    Covidien
    ICU Medical
    Kane Biotech*
    KCI
    Next Science*
    Semprus Biosciences*
    Steris
    W.L. Gore

    Biofilm in oilfield pipeline system

    Cooling tower biofilms

    Health-related biofilms

    Chronic wounds

    Ellen Swogger, Garth James et al., MSU-CBE, 2005. Cover image on Wound Repair and Regeneration
    16(1), January-February 2008, ISSN 1067-1927, Blackwell Publishing

    Elbow prosthesis biofilm

    Urinary catheter encrustation

    Morris, N.S. and Stickler, D.J. (2001) BJU Int 88:192

    Dental caries

    Where ever moisture and nutrients are


    present, there will be biofilms!

    Biofilm in a water distribution system

    Biofilm in a water distribution system

    Why not just eradicate them with


    antimicrobial agents?

    Biofilms are more tolerant of


    antimicrobial agents than are free bugs
    Glutaraldehyde (50 mg/l)
    against P. aeruginosa

    Log Reduction

    Biofilm

    Planktonic
    3

    200

    400

    Time (min)

    600

    800

    Biofilms are more tolerant of


    antimicrobial agents than are free bugs
    2

    control

    biofilm

    Log (X/Xo)

    0
    -1
    -2

    Tobramycin
    against P. aeruginosa

    -3
    -4

    planktonic

    -5
    -6
    0

    25

    50
    Time (h)

    75

    100

    Biofilm Tolerance
    of Antimicrobial
    Antimicrobial
    tolerance
    can be Agents
    understood in light of three concepts

    1. Slow penetration

    Measure penetration with


    microelectrodes

    Hydrogen
    Peroxide
    Hydrogen
    PeroxideConcentration
    Concentration (mM)(mM)

    H2O2 fails to penetrate the biofilm


    60

    chloride

    bulkfluid
    fluid
    Bulk

    50
    40
    30

    H2O2

    20

    catalase

    hypochlorite

    10
    0

    H2O + 1/2O2

    base
    biofilm
    Base
    ofofbiofilm
    0

    600

    1200

    1800

    2400

    Time (sec)
    Time
    (seconds)

    3000

    3600

    Chloride ions penetrate readily but


    hypochlorite ions do not
    Relative Penetration

    1.0

    Cl-

    0.8
    0.6
    0.4
    0.2
    0.0

    OCl0

    200

    400
    600
    Time (sec)

    800

    Hypochlorite penetrates biofilm slowly


    1200
    1000
    800

    600

    5460 s

    400
    200
    0

    1511 s
    47 s

    200 400 600 800 1000 1200

    Depth (microns)

    2. Altered Microenvironment

    Oxygen in P. aeruginosa Biofilm


    Oxygen Concentration (mg/l)

    7
    sterile control

    6
    5
    4
    3
    2
    1
    0
    -50

    untreated

    50

    100

    Depth ( mm)

    150

    200

    GFP Expression in P. aeruginosa Biofilm

    DNA Replication Pattern Biofilm

    3. Stress Response

    Catalase Induction in P. aeruginosa


    Biofilm
    65

    biofilm + H2O2

    60

    katB-lacZ

    55
    50
    45
    40

    planktonic + H2O2

    35
    30

    biofilm - untreated

    25
    0

    10

    20

    30

    Time (min)

    40

    50

    60

    Imaging antimicrobial action

    S. epidermidis
    biofilm staining
    with Calcein AM
    green (CAM)
    Time: 1 hr
    Vel = 6 cm/sec

    Imaging antimicrobial action

    CAM-stained
    S. epidermidis
    biofilm treated
    with 10 mg/L
    sodium
    hypochlorite
    Time: 1 hr
    Vel = 6 cm/sec
    W. Davison

    Imaging antimicrobial action

    CAM-stained
    S. epidermidis
    biofilm treated
    with 50 mg/L
    sodium
    hypochlorite
    Time: 1 hr
    Vel = 6 cm/sec
    W. Davison

    Options for Microbial Control

    Biofilm control in industrial systems


    Regular cleaning and antimicrobial dosing
    Antimicrobial access to the biofilm

    Contact time
    Materials Compatibility
    Measuring successful treatment

    Contents of these slides


    were provided by faculty,
    staff, and students in the
    Center for Biofilm
    Engineering at Montana
    State University
    www.biofilm.montana.edu
    Thank you for your
    attention!

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