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Microbiology

Microbial Growth

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

Log Phase 3. Death Phase .Phases of Growth • • • • 1. Lag Phase 2. Stationary Phase 4.

1.evtl.cells adapt to new medium and factors .g.no immediate increase in cell number and mass occurs . type of medium.length of lag phase dependent on different factors: e. presence or absence of important co-factors or supplements . temperature.temperature adaptation . age of cell culture. new enzymes and metabolic pathways are induced . Lag Phase .

Log Phase • Rapid cell growth (exponential growth) • bacteria are dividing and doubling in number at constant rate • microbes are sensitive to adverse conditions • antibiotics • anti-microbial agents - .2.

2.lgN0 / lg2 Division rate or mean growth rate constant k: k = n / t = lgN . Log Phase the cell count N of a cell population with a number of N0 cells (at the beginning of the inoculation) and after n cell divisions can be calculated using following formula: N(n) = N0 2n Number of cell divisions n: n = lgN .lgN0 / lg2 (t .t0) --> number of generations per unit of time Mean generation time or Doubling time g: g = 1/ k .

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increased peptidoglucan cross-linking.3. amino acids) • Energy-consuming production of starvation proteins. DNA-protecting proteins (Dps). water. glucose. carbon source (e. Stationary Phase • population growth ceases and growth curve becomes horizontal • Death rate = rate of reproduction • bacterial population density reaches maximum number: usually 109 cells / ml • cells begin to encounter environmental stress • lack of nutrients.g. stress-response proteins • not enough space • metabolic wastes • Oxygen down • pH down (becomes acid) • Endospores would form now . chaperons. e.g.

4. water or nutrients • space • accumulation of metabolic wastes • lack of oxygen • changes in pH • temperature • decline in number of viable cells due to build-up of toxic metabolic end products • cells loose ability to reproduce and die off. Death Phase • Death rate > rate of reproduction • Due to limiting factors in the environment • Lack of food. some produce endospores death rate may be exponential .

pH. Osmotic pressure) • Chemical (nutrient/elements.refers to the # of cells.Microbial Growth . oxygen) . not the size of the cells • Requirements for Growth • Physical (Temperature.

loving microbes) • range • thermophiles • range 20 C .40 C (heat loving microbes) 40 C .20 C • mesophiles (moderate temp.100 C .Physical Requirements • Temperature • psychrophiles (cold loving microbes ) • range 0 C .

0 • many foods.pH 7.5 .5 • Very few can grow at below pH 4.pH • Most bacteria grow between pH 6. pickles. and cheeses are preserved from spoilage by acids produced during fermentation . such as sauerkraut.

Osmotic Pressure • Microbes obtain almost all their nutrients in solution from surrounding water • Tonicity • isotonic • hypertonic • hypotonic .

V. Ca. S.Chemical Requirements • Macro elements (there are ten): C. • micro. O. P. N. . Ni. Zn. K. Cu. Mg and Fe. Co. B. Cl. Mo. Na. H. Si and W.or trace-elements f(or optimum growth): Mn. Se.

Microaerophilic. . Obligate Anaerobes. Facultative Anaerobes.Oxygen • Bacteria can be classified base on their oxygen requirements Obligate Aerobes.

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 • Catalase • 2 H2O2 --------> 2 H2O + O2 • Obligate Anaerobes lack: • Superoxide dismutase ( SOD ) • Catalase .

tryptone or meat . Tryptic soy broth 2.a minimum medium is a solution which contains just enough of the chemicals and compounds to allow good growth of the specific microorganism • Complex media: nutrient solution which contains defined amounts of chemically not defined extracts. MacConkey agar . peptone.commonly used complex media in microbiology are: 1.g. extracts from yeast. Luria Bertani (LB) broth 5. Nutrient broth 3.Growth Media • Synthetic media : solution which is comprised of defined amounts chemical ingrediences and compounds . Peptone yeast extract broth 4.they are often the choice or only option if the exact growth conditions and requirements of a given microorganism are not known . e.

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Tract infections are caused by Gram (-) bacteria .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 caused by Gram (-) bacteria .I.Selective Media • EMB (Eosin Methylene Blue) • dyes inhibit Gram (+) bacteria • selects for Gram (-) bacteria • G.

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 Beta Hemolytic Streptococci .

(7.5%) inhibits most bacteria • Contains sugar mannitol • pH Indicator (Turns Yellow when acid) • MacConkey’s Agar • used to identify Salmonella • Bile salts and crystal violet (inhibits Gram (+) bacteria) • Contains lactose • pH Indicator .Selective and Differential Media • Mannitol Salt Agar • used to identify Staphylococcus aureus • High salt conc.

Thank you! .

Complex Media • exact chemical composition is not known • most bacteria and fungi are grown with this .Culture Media • 1. Chemically Defined • the exact chemical composition is known • used to grow fastidious organisms • 2.

Agar Shake . Reducing Media b.Special Culture Techniques • 1. Anaerobic Bacteria • • • • a. Anaerobic Container c. Agar Stab d.

Special Culture Techniques • 2. 0.03 % CO2 .3 to 0. Microaerophilic Bacteria • grow best under reduced O2 levels and increased CO2 levels Normal Atmosphere 21 % O2.

Microaerophilic Bacteria A. Candle Jar 16 % O2 4% CO2 .

Microaerophilic Bacteria B. CO2 Generating Packet .

Enumeration of Bacteria • Turbid culture .10 million bacterial cells per ml • Serial Dilution .

Petroff-Hauser Counting Chamber • 2. .• 1.