I.
MICROBIAL GROWTH
1. Meaning of Microbial Growth
Microbial growth = increase in number of cells, not cell size.
Growth occurs through binary fission (one cell divides into two).
2. Temperature Requirements
Psychrophiles – grow in very cold temps (0–15°C).
Psychrotrophs – grow at 0°C but prefer 20–30°C; cause food spoilage.
Mesophiles – grow at 25–40°C; includes most human pathogens.
Thermophiles – survive at 50–60°C.
Hyperthermophiles – thrive above 80°C, often in hot springs.
3. pH Requirements
Most bacteria prefer pH 6.5–7.5.
Acidophiles – grow in acidic conditions.
Neutrophiles – prefer neutral pH.
Alkaliphiles – prefer basic pH.
4. Osmotic Pressure
Halophiles – tolerate or require high salt concentrations.
5. Chemical Requirements for Growth
Carbon – makes up about 50% of bacterial dry weight; needed for all organic compounds.
Nitrogen – used to build proteins; usually obtained from decomposed proteins.
Other important elements: sulfur, phosphorus, trace elements.
6. Oxygen Requirements
Obligate aerobes – require oxygen.
Obligate anaerobes – cannot grow in presence of oxygen.
Facultative anaerobes – grow with or without O₂.
Aerotolerant anaerobes – tolerate oxygen but don’t use it.
7. Toxic Forms of Oxygen
Superoxide radical (O₂⁻) – very toxic; must be neutralized by superoxide dismutase (SOD).
Other types: hydrogen peroxide, singlet oxygen, hydroxyl radicals.
8. Biofilms
Communities of microbes embedded in slime layers.
Found on catheters, drains, teeth (plaque), etc.
II. CULTURING MICROORGANISMS
1. Media Types
Medium – nutrients for microbial growth.
Chemically defined medium – exact chemical composition known.
Complex medium – contains extracts (exact composition unknown).
Selective medium – suppresses unwanted microbes; promotes desired ones.
Differential medium – distinguishes colonies by color changes.
Enrichment medium – encourages growth of rare microbes.
2. Culture Techniques
Inoculum – microbes introduced into a medium.
Agar – solidifying agent.
Streak plate method – used to get pure cultures.
Colony or CFU – population of cells from one cell.
3. Preserving Cultures
Deep-freezing: −50° to −95°C
Lyophilization (freeze-drying): frozen and dehydrated in a vacuum
III. BACTERIAL GROWTH CURVE
4 Phases:
Lag phase – little or no cell division.
Log (exponential) phase – rapid cell division.
Stationary phase – growth slows; nutrients depleted.
Death phase – cells die.
Generation time
Time required for a cell to divide.
Measuring Growth
Direct methods: plate counts.
Indirect methods: turbidity using spectrophotometer.
IV. MICROSCOPY
1. Size Units
Bacteria – usually in micrometers (µm).
Viruses – measured in nanometers (nm).
2. Important People
Anton van Leeuwenhoek – first observed microorganisms (“animalcules”); examined dental plaque.
Robert Hooke – coined the word “cell” while looking at cork.
Hans & Zacharias Janssen – created the first compound microscope.
Ernst Ruska & Max Knoll – pioneered electron microscopy.
Ivanovsky & Beijerinck – worked on Tobacco Mosaic Virus.
3. Types of Microscopes
Light microscope – for bacteria.
Electron microscope – for viruses.
4. Properties of Microscopes
Magnification – how large the image becomes.
Resolution – ability to distinguish two close objects.
Contrast – visibility of specimen vs. background.
5. Measuring Microorganisms
Stage micrometer – standard for calibration.
Ocular micrometer – measuring scale inside the eyepiece (must be calibrated first).
6. Historical Contributions
Greeks & Romans – used water lenses.
Galileo – improved lens systems.
Joseph Lister – reduced distortions in lenses.
V. MICROBIAL CONTROL & STERILIZATION
Pasteurization – reduces microbial load, does not sterilize.
HTST pasteurization – 72°C for 15 seconds.
Chlorine – forms hypochlorous acid in water.
Asepsis – absence of significant contamination.
Hydrogen peroxide – oxidizing agent inactivated by catalase.
Antiseptic – used on living tissue.
Disinfectant – used on non-living surfaces.
Alcohol (70–90%) – most effective range.
Sterilization – removes all microbial life, including endospores.
Sanitization – reduces microbes on utensils to safe levels.