MICROBIOLOGY

Micro – means very small

bio – refers to living organisms
Logy – means the study of

MICROBIOLOGY
-Study of very small living organism.
These includes: Bacteria, algae, protozoa, fungi, viruses (often called
microbes, single-celled organisms. You cannot see microorganisms
without the aid of a microscope.
Pathogens
Disease causing organisms.
Only a small percentage of microbes are pathogens
Some cause diseases only if they accidentally invade the “wrong place”
such as when the host’s resistance is low and growth conditions are
right.
History of Microbiology
Johannes Janssen
-developed the use of 2 or more lenses and increase the magnification.
Robert Hooke
-marked the beginning of cell theory that cell living things are
composed of cells.
Anton Van Leeuwenhoek
-Father of Microbiology
-described bacteria, protozoa, sperm and blood cells and called them
animalcules.
Louis Pasteur
-described the process of fermentation and pasteurization.
Fermentation – converts sugar to alcohol (in the absence of air).
Pasteurization - commonly used to kill potentially harmful bacteria in
milk and dairy products.
Laurent Lavosier
-showed the importance of oxygen to life.
John Needham
-He assumed that microbes spontaneously form from fluids.
John Lister
-Father of aseptic surgery
Robert Koch
-Germ theory of diseases
Edward Jenner
-Father of Immunology
 Alexander Fleming
- Discovered penicillin
Emil von Behring
-developed diphtheria antitoxin.
Paul Erlich
-developed theories of immunity
Selman Waksman
-discovered streptomycin
Rene Dubos
-discovered gramicidin and thyrocidin
John Tyndall
-disproved the theory of spontaneous generation and proved that life
must arise from pre-existing life (theory of biogenesis) 1st proposed by
Rudolf Virchow.
-found bacterial fragments called “endospores” that were not destroyed
by boiling and that germinated into reproducing vegetative bacteria
which led to the development of the fractional sterilization process
(tyndallization)
Carolus Linnaeus
-Established the scientific nomenclature.
Various Types of Microscopes
Light Microscope
-Single lens magnifying glass usually magnifies the image of an object
from 3-20x the actual size.
-Today, a Compound Brightfield Microscope with two lens system and
visible light source that passes through the specimen and the lenses to
the observer’s eye.
magnify – 40-1000x
magnifying lens- x4, 10, 100
Transmission Electron Microscope
-specimen can be viewed on screen.
-Excellent resolution
-allows examination of cellular ultrastructure and viruses
-specimen is non-living
-image is two-dimensional
Scanning Electron Microscope
-For observing surfaces and the 3D image of an object.
-Specimen can be viewed on screen.
-Useful in examining surface structures of cells and viruses.
Darkfield Microscope
-background is dark
-unstained organism can be seen
-useful for examining spirochetes
-slightly more difficult to operate than brightfield.
Phase Contrast Microscope
-can observe dense structures in living prokaryotic and eukaryotic
microorganisms.
Fluorescence Microscope
-fluorescent dye attached to organism
-to detect microorganism in cells, tissue and clinical specimen.
Units of Measurement:
Meter – basic unit of the metric system
Micron – has been replaced by the term micrometer (um), millimicron (mu)
Nanometer (nm)
II. Types of Microorganisms:
Modern classification of living organisms: Objective to establish
relationship among microorganisms.
I. Superkingdom
II. Kingdom
III. Division/ Phylum
IV. Order
V. Family
VI. Genus
VII. Species
Superkingdom Prokaryotes – lack true cell nucleus
Eukaryotes – with true nucleus

Prokaryotes
- The genetic material is in mixture with the cytoplasm
- Bacteria possess DNA and RNA
- Average size ranges 0.5-2um
- Divide by simple division (binary fission)
- All bacteria are prokaryotes
Eukaryotes
-contain genetic material which is separated from the cytoplasm
Bacteria
– single celled organism
-larger than viruses
Fungi
-obtain nourishment by absorbing solution of organic material from their environment (soil, sea,
water)
-more advanced cell structure than bacteria and viruses
-can reproduce by mitosis or meiosis.
Viruses
- Smallest infectious agent with pathogenic potential
- Simply a lifeless arrangement of molecules that can pass through filters
- Less than 1 u
- Best observed in Electron Microscopy.
Algae
- Commonly green, red, golden brown, yellow green and often referred to have chlorophyll.
Parts of a BACTERIAL CELL
1. Cell wall
-The external structure
-Provides rigidity and protection
Eukaryotic cell wall- consist mainly of cellulose but may contain
pectin, lignin, chitin and some mineral salts.
Prokaryotic cell wall- chemical composition (peptidoglycan or
murein)
gram (+) – techoic acid (thick layer)
gram (-) - (thinner layer)lipopolysaccharide, phospholipid,
lipoprotein- site of cell’s permeability
a. Permeability – bacterial cell is permeable to water and maintains an
osmotic pressure between the cytoplasm and the surrounding
medium.
b. Plasmolysis – bacterial cell wall when grown in a saline solution
which is more dense than the concentration of the protoplasm. It
will causes shrinkage because the water passes out forming the so
called shadow form.
c. Plasmoptysis – when bacterial cell is placed in distilled water of low
concentration, water passes in the cell wall (swells/ burst)
2. Cell membrane/ plasma membrane/ cytoplasmic membrane
-site of respiratory enzyme and act as osmotic barrier.
-regulate the passage of nutrients and secretions from outside to the
inside of the cell.
-consist of a watery sap packed with large number of small granules
called ribosomes and few convoluted membranous bodies called
mesosomes where many enzymes are concentrated so respiration
takes place.
3. Capsules
-the outermost layer of a bacterial cell.
-thickened, slimy or gelatinous layer
-delay the digestion of bacteria by phagocytes
-usually associated with virulence
-consist of polysaccharide complex with lipids and proteins
-can be detected and demonstrated by negative staining (nigrosine or india
ink).
Example of organisms with capsule:
S. pneumoniae N. meningitides
K. pneumoniae N. gonorrhea
H. Influenzae B. anthracis
Capsulated organisms are pathogenic but not flagellated
4. Nucleus
-control and integrates the function of the entire cell.
-contain 1 or more chromosomes and number of genes containing DNA.
5. Metachromatic granules
-made up of polymetaphosphates
-represents accumulation of food reserves
Ex. Babes Ernst bodies – C. diptheriae
6. Pili or fimbriae
-tiny hair-like filaments found in some bacteria usually produced by flagellated gram
(-) bacteria (e.g E. coli)
-facilitates adherence of microorganisms to tissues and therefore contribute to
virulence.
-perform some sexual functions which is conjugation.
7. Spores
-Resistant to heat, drying and most chemicals.
8. Flagella
-slender, whiplike structures which serves as means of locomotion or
motility of the organism.
CLASSIFICATION OF BACTERIA ACCORDING TO THE # OF FLAGELLA AND
LOCATION
a. Atrichous – No flagella
b. Monotrichous – One flagellum at one end
c. Amphitrichous – one flagellum on both end
d. Lopotrichous – group or tuft of flagella on one or both end of the
organism.
e. Peritrichous – flagella surrounding the bacterial body.
CLASSIFICATION OF
BACTERIA:
I. Morphology/ Shapes of Bacteria:
1. Cocci (coccus) – round or spherical organism
2. Bacilli (bacillus) – rod-shaped organism
3. Spirilli (spirillum) – curved or spiral in shape
a. Vibrio – straight rod with single rigid curve
b. Spirillum – rigid helical rod
c. Spirochete – flexus helical rod
4. Pleomorphic – vary in size and shape (coccobacilli)
II. Arrangement:
1. Grapelike clusters – e.g. Staphylococcus
2. Chain – Streptococci, Streptobacilli
3. In pairs – diplococcic, diplobacilli, coccobacilli
4. tetrads – packets of four eg. Micrococcus, Peptococcus
5. Sarcinae – packets of eight
6. Palisade – tend to place themselves (side by side) resembling a pack of
cigarettes eg. C. diptheriae
7. Chinese character arrangement – snapping organisms tend to bend at
point of division losing their characteristic shape because of advance
growth conditions resembling a Chinese letter.
III. Size
Average size – 0.5-2.0 um
Haemophilus – smallest pathogenic organism (.2-.5um)
B. anthracis – largest pathogenic organism (1x3 – 10um)
Control of Microbial Growth
Reasons why microbial growth must be controlled:
1. To prevent infectious disease in harming animals and plants.
2. To preserve food.
3. To prvent contaminating microbes from interfering with certain
industrial processes.
4. To prevent contamination of pure culture.
STERILIZATION
Complete removal or destruction of all living organisms.
It is affected by physical or chemical means.

Physical Method of Destruction:

A. Moist Heat Method
◦ Principle: It kills the microorganisms by coagulating the protein.
1.Boiling – used to sterilize surgical instruments, needles, syringes, glass petri
dishes (100°C for 15-30 minutes)
2. Autoclave – most effective
-steam under pressure, temperature employed 121°C
-time of exposure: 15-30 minutes
-pressure 15 psi
3. Arnold’s sterilizer
Tyndallization/Flowing Steam/Fractional/ Intermittent Sterilization
- 3 consecutive days at 100°C for 30 min
-For media such as sugars, gelatin, milk etc.
4. Inspissation
- 75-80°C
- thickening thus evaporations
- 2 hrs for 3 consecutive day
- usually used to sterilize high protein containing media that cannot
withstand the high temperature of the autoclave.
5. Pasteurization
- partial sterilization by heat
- used to sterilize milk and milk products
Methods:
1. LTH – Low temperature holding - 62°C for min known as Batch
Method.
2. HTST – High temperature short time - 72°C for 15 secs known as
Flash Method.
B. Dry Heat Method
◦ Principle: Denaturation of protein by oxidation
1. Flaming mouth of tubes, wireloops and forceps.
2. Oven temp. must be 160-180°C with a time exposure of 1 hr and
30 mins to 2 hrs.
(HOT AIR STERILIZER)
-to sterilize glasswares, certain metals
3. Incineration – burning material to ashes
300-400°C
4. Cremation – burning human bodies to ashes to control
communicable diseases.
Cold temperature:
Refrigeration – ref temp. w/n 3 days (e.g. T. pallidum are killed) 2-8°C

C. Radiation – makes use of UV light of rays/ genetic composition is altered.

D. Trituration or Grinding
E. Mechanical Shaking or Agitation
F. Use of Ultrasonics or Vibrations at high frequency
G. Filtration – used to sterilize heat labile substances such as serum,
plasma and certain carbohydrate solution (urea, broth, sugar
fermentation broth)
H. Freeze Drying/ Lyophilization
I. Ultracentrifugation