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    Microbial Growth: Dr. Husni Samadin

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    Nur Habib Al-Bangkawi
    Microbial growth refers to the increase in number of cells rather than size. Requirements for growth include physical factors like temperature between 0-100°C and pH between 6.5-7.5, as well as chemical requirements like macro/micronutrients. Bacteria are classified based on oxygen requirements as obligate aerobes, obligate anaerobes, facultative aerobes, or microaerophilic. Culture media can be chemically defined or complex, and special techniques are used to culture anaerobic and microaerophilic bacteria. Selective and differential media allow isolation and identification of specific bacteria. Bacterial growth occurs through binary fission in four phases: lag, log, stationary,

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    Microbial Growth: Dr. Husni Samadin

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    Nur Habib Al-Bangkawi
    31 views37 pages
    Microbial growth refers to the increase in number of cells rather than size. Requirements for growth include physical factors like temperature between 0-100°C and pH between 6.5-7.5, as well as chemical requirements like macro/micronutrients. Bacteria are classified based on oxygen requirements as obligate aerobes, obligate anaerobes, facultative aerobes, or microaerophilic. Culture media can be chemically defined or complex, and special techniques are used to culture anaerobic and microaerophilic bacteria. Selective and differential media allow isolation and identification of specific bacteria. Bacterial growth occurs through binary fission in four phases: lag, log, stationary,

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    Chapter 6 2008

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    Microbial growth refers to the increase in number of cells rather than size. Requirements for growth include physical factors like temperature between 0-100°C and pH between 6.5-7.5, as well as chemical requirements like macro/micronutrients. Bacteria are classified based on oxygen requirements as obligate aerobes, obligate anaerobes, facultative aerobes, or microaerophilic. Culture media can be chemically defined or complex, and special techniques are used to culture anaerobic and microaerophilic bacteria. Selective and differential media allow isolation and identification of specific bacteria. Bacterial growth occurs through binary fission in four phases: lag, log, stationary,

    Copyright:

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    Download as ppt, pdf, or txt
    31 views37 pages

    Microbial Growth: Dr. Husni Samadin

    Uploaded by

    Nur Habib Al-Bangkawi
    Microbial growth refers to the increase in number of cells rather than size. Requirements for growth include physical factors like temperature between 0-100°C and pH between 6.5-7.5, as well as chemical requirements like macro/micronutrients. Bacteria are classified based on oxygen requirements as obligate aerobes, obligate anaerobes, facultative aerobes, or microaerophilic. Culture media can be chemically defined or complex, and special techniques are used to culture anaerobic and microaerophilic bacteria. Selective and differential media allow isolation and identification of specific bacteria. Bacterial growth occurs through binary fission in four phases: lag, log, stationary,

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    of 37

    Microbial Growth

    Dr. Husni Samadin

    Microbial Growth - refers to the # of


    cells, not the size of the cells

    Requirements for Growth


    Physical Chemical

    Physical Requirements

    Temperature
    psychrophiles (cold loving microbes )
    range range 0 C - 20 C 20 C - 40 C

    mesophiles (moderate temp. loving microbes)


    thermophiles
    range

    (heat loving microbes)


    40 C - 100 C

    pH

    Most bacteria grow between pH 6.5 - pH 7.5


    Very few can grow at below pH 4.0
    many foods, such as sauerkraut, pickles, and cheeses are preserved from spoilage by acids produced during fermentation

    Osmotic Pressure

    Microbes obtain almost all their nutrients in solution from surrounding water
    Tonicity
    isotonic hypertonic hypotonic

    Cells

    Chemical Requirements

    Macro & Micro Elements

    C HOPKINS CaFe Mg NaCl

    Oxygen

    Bacteria can be classified base on their oxygen requirements

    1. Obligate Aerobes

    2. Obligate Anaerobes

    3. Facultative Aerobes
    Facultative Anaerobes

    4. Microaerophilic

    Oxygen is lethal to some organisms

    All organisms produce superoxide ( O2-)


    Superoxide is toxic to cells (steals electrons) Superoxide must be neutralized

    Superoxide dismutase

    O2- + O2- + 2 H+ -------> H2O2 + O2

    Hydrogen peroxide is also toxic to cells and it must be neutralized

    Catalase

    2 H2O2 --------> 2 H2O + O2 Obligate Anaerobes lack:


    Superoxide dismutase ( SOD ) Catalase

    Culture Media

    1. Chemically Defined
    the exact chemical composition is known used to grow fastidious organisms

    2. Complex Media
    exact chemical composition is not known most bacteria and fungi are grown with this

    Special Culture Techniques

    1. Anaerobic Bacteria
    a. Reducing Media b. Anaerobic Container c. Agar Stab d. Agar Shake

    Special Culture Techniques

    2. Microaerophilic Bacteria
    grow best under reduced O2 levels and increased CO2 levels Normal Atmosphere 21 % O2 .3 to .03 % CO2

    Microaerophilic Bacteria

    A. Candle Jar

    16 % 4%

    O2 CO2

    Microaerophilic Bacteria
    B. CO2 Generating Packet

    Selective Media

    Inhibits the growth of some bacteria while selecting for the growth of others Example:
    Brilliant Green Agar
    dyes inhibit the growth of Gram (+) bacteria selects for Gram (-) bacteria Most G.I. Tract infections are caused by Gram (-) bacteria

    Selective Media

    EMB (Eosin Methylene Blue)


    dyes inhibit Gram (+) bacteria selects for Gram (-) bacteria G.I. Tract infections caused by Gram (-) bacteria

    Differential Media

    Differentiates between different organisms growing on the same plate Example:


    Blood Agar Plates (TSA with 5% sheep blood)
    used to differentiate different types of Streptococci

    Alpha Hemolytic Streptococci

    Incomplete lysis of RBCs

    Beta Hemolytic Streptococci

    Complete lysis of RBCs

    Gamma Hemolytic Streptococci

    No lysis of RBCs

    Selective and Differential Media

    Mannitol Salt Agar


    used to identify Staphylococcus aureus

    Mannitol Salt Agar


    High salt conc. (7.5%) inhibits most bacteria sugar Mannitol pH Indicator (Turns Yellow when acid)

    Selective and Differential Media

    MacConkeys Agar
    used to identify Salmonella

    MacConkeys Agar
    Bile salts and crystal violet (inhibits Gram (+) bacteria) lactose pH Indicator

    Many Gram (-) enteric non-pathogenic bacteria can ferment lactose, Salmonella can not

    Bacterial Growth - increase in the # of


    cells

    Binary Fission
    Generation Time (Doubling Time)
    time required for a cell to divide most about 1 Hr. To 3 Hrs.
    E. coli - 20 minutes Mycobacterium tuberculosis - 24 Hrs.

    Binary Fission - unchecked

    E. coli - generation time of 20 min. 20 generations (about 7 hrs.)


    1 million cells

    30 generations ( about 10 hrs.)


    1 billion cells

    72 generations ( about 24 hrs.)


    1 x 1021
    1,000,000,000,000,000,000,000 cells

    Limiting factors in the environment

    Lack of food, water or nutrients space accumulation of metabolic wastes lack of oxygen changes in pH temperature

    Phases of Growth

    4 Phases
    1. Lag Phase 2. Log Phase 3. Stationary Phase 4. Death Phase

    1. Lag Phase

    Bacteria are first introduced into an environment or media Bacteria are checking out their surroundings cells are very active metabolically # of cells changes very little 1 hour to several days

    2. Log Phase

    Rapid cell growth (exponential growth) population doubles every generation microbes are sensitive to adverse conditions
    antibiotics anti-microbial agents

    3. Stationary Phase

    Death rate = rate of reproduction cells begin to encounter environmental stress


    lack of nutrients lack of water not enough space metabolic wastes oxygen pH

    Endospores would form now

    4. Death Phase

    Death rate > rate of reproduction Due to limiting factors in the environment

    Enumeration of Bacteria

    Turbid culture Serial Dilution

    10 million bacterial cells per ml

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