STM 211

INTRODUCTION TO MICROBIOLOGY

COURSE OUTLINE
WEEKS 1-2 The Scope of Microbiology (Scientists, roles of Microbiology)
WEEK 3 Principle of Microscopy (Types of Microscope, Applications and Staining
Techniques).
WEEK 4 Characteristics of Microorganisms (Morphology/ Biochemical Basis for
Classification)
WEEK 5-6 Nutritional Requirements of Microorganisms
WEEK 7 Breakdown of food molecules by Microorganisms, Microbial Growth
Curve
WEEK 8 Types and Composition of Culture Media
WEEK 9-11 Pure / Mixed Cultures, Culture Characteristics, Maintaining Pure Cultures
WEEK 12-14 Controlling Microorganisms
WEEK 15 Transporting Culture Samples

INTRODUCTION TO MICROBIOLOGY
Microbiology is the study of microorganisms. These organisms are microscopic size and can be
viewed by the microscope. The term microbiology was given by French chemist Louis Pasteur
(1822-95).
Many scientists contributed to the field of Microbiology. Notable among them are;

1. Robert Hooke
A 17th-century English scientist, was the first to use a lens to observe the smallest unit of tissues
he called “cells.” Soon after, the Dutch amateur biologist Antoni van Leeuwenhoek observed
what he called “animalcules” with the use of his homemade microscopes.
2. Antoni van Leeuwenhoek (1632-1723)
Of Delft, Holland (Netherland) was the first person to observe and accurately describe
microorganisms (bacteria and protozoa) called ‘animalcules’ (little animals) in 1676.
 Actually he was a Dutch linen merchant but spent much of his spare time constructing simple
microscopes composed of double convex lenses held between two silver plates. He constructed
over 250 small powerful microscopes that could magnify around 50-300 times.
Leeuwenhoek was the first person to produce precise and correct descriptions of bacteria and
protozoa using a microscope he made himself. Because of this extraordinary contribution to
microbiology, Antoni van Leeuwenhoek is considered as the “Father of microbiology”.

The Golden age of microbiology began with the work of Louis Pasteur and Robert Koch who
had their own research institute. More important there was an acceptance of their work by the
scientific community throughout the world and a willingness to continue and expand the work.
During this period, we see the real beginning of microbiology as a discipline of biology.
3. The concept of spontaneous generation was finally put to rest by the French
chemist Louis Pasteur in an inspired set of experiments involving a goose necked flask. When
he boiled broth in a flask with a straight neck and left it exposed to air, organisms grew. When he
did this with his goose-necked flask, nothing grew. The S-shape of this second flask trapped dust
particles from the air, preventing them from reaching the broth. By showing that he could allow
air to get into the flask but not the particles in the air, Pasteur proved that it was the organisms in
the dust that were growing in the broth.
 Pasteur, thus in 1858 finally resolved the controversy of spontaneous generation versus
biogenesis and proved that microorganisms are not spontaneously generated from inanimate
matter but arise from other microorganisms.
 He also found that fermentation of fruits and grains, resulting in alcohol, was brought about by
microbes and also determined that bacteria were responsible for the spoilage of wine during
fermentation. Pasteur in 1862 suggested that mild heating at 62.8°C (145°F) for 30 minutes
rather than boiling was enough to destroy the undesirable organisms without ruining the taste
of the product, the process was called Pasteurization. Pasteurization was introduced into the
United States on a commercial basis in 1892. His work led to the development of the germ
theory of disease.
 Louis Pasteur is known as the “Father of Modern Microbiology / Father of Bacteriology.
He has many contributions to microbiology:
1. He has proposed the principles of fermentation for the preservation of food.
2. He introduced sterilization techniques and developed steam sterilizers, hot air oven, and
autoclave.
3. He described the method of pasteurization of milk.
4. He had also contributed for designing the vaccines against several diseases such as anthrax,
fowl cholera, and rabies
5. He disproved the theory of spontaneous generation of disease and postulated the ‘germ theory
of disease’. He stated that disease cannot be caused by bad air or vapor, but it is produced by
the microorganisms present in the air.
6. Liquid media concept- He used nutrient broth to grow microorganisms.
7. He was the founder of the Pasteur Institute, Paris.
 4. Around the same time that Pasteur was doing his experiments, a doctor named Robert
Koch was working on finding the causes of some very nasty animal diseases (first anthrax, and
then tuberculosis). He gave the first direct demonstration of the role of bacteria in causing
disease. He was a German physician who first of all isolated anthrax bacillus (Bacillus anthracis,
the cause of anthrax) in 1876. He perfected the technique of isolating bacteria in pure culture. He
also introduced the use of solid culture media in 1881 by using gelatin as a solidifying agent. In
1882 he discovered Mycobacterium tuberculosis. He proposed Koch postulate which were
published in 1884 and are the corner stone of the germ theory of diseases and are still in use
today to prove the etiology (specific cause) of an infectious disease.
Koch’s four postulates are:
 The organism causing the disease can be found in sick individuals but not in healthy ones.
 The organism can be isolated and grown in pure culture.
 The organism must cause the disease when it is introduced into a healthy animal.
 The organism must be recovered from the infected animal and shown to be the same as the
organism that was introduced.
5. Lord Joseph Lister (1827-1912): A famous English surgeon is known for his notable
contribution to the antiseptic treatment for the prevention and cure of wound infections. Lister
concluded that wound infections too were due to microorganisms. In 1867, he developed a
system of antiseptic surgery designed to prevent microorganisms from entering wounds by the
application of phenol on surgical dressings and at times it was sprayed over the surgical areas.
He also devised a method to destroy microorganisms in the operation theatre by spraying a fine
mist of carbolic acid into the air, thus producing an antiseptic environment. Thus, Joseph Lister
was the first to introduce aseptic techniques for control of microbes by the use of physical and
chemical agents which are still in use today. Because of this notable contribution, Joseph Lister
is known as the Father of Antiseptic surgery.
6. Alexander Fleming (1881-1955): He discovered the antibiotic penicillin.
He was a British Bacteriologist. He observed a mold (Penicillium notatum) growing on a plate
of Staphylococcus aureus. The growth of Staphylococcus aureus around the mold colony
was inhibited.
 7. Ferdinard Cohn
He was a trained botanist and became an excellent microscopist. His interests in microscopy led
him to the study of unicellular plants. He discovered the important group of bacteria that form
endospores. He laid the groundwork for a system of bacterial classification. He founded a major
scientific journal of plant and microbial biology.
Applications of Microbiology
1. Bioremediation
Living organisms are used to degrade environmental pollutants. This process is called
Bioremediation. Bacteria are being used to degrade oil, assist in the cleaning of oil spills and
treat radioactive wastes. They are also useful in Bio-treatment of wastes (the use of living
organisms in the treatment of waste).
2. Food Production
Many dairy products depend on microbial transformations including the fermentations that yield
cheeses, yoghurt and buttermilk. Baked goods and alcoholic beverages rely on the fermentative
activities of yeast, generating carbon dioxide to raise the dough and alcohol as key ingredient
respectively.
3. Agriculture
Microorganisms are essential for the digestive process in ruminant animals such as cattle and
sheep. Microorganisms play key roles in the cycling of important nutrients in plant nutrition
particularly those of carbon, nitrogen and Sulphur. Apart from their benefits, microorganisms
cause diseases of plant and animals affecting the economic value of their products.
4. Pharmaceutical industry
The most important contribution of microbiology to this industry is the development of
antibiotics. All antibiotics were originally the products of microbial metabolism; however, the
recent genetic manipulations have enabled the production of more enhanced drugs.
5. Medicine
The discipline of Microbiology helps to recognize, isolate, diagnose and treat pathogenic
microorganisms and also produces beneficial organisms such as yeasts and some antibiotics.
Microbiology also develops new methods for preventing illness.

PRINCIPLES OF MICROSCOPY
Visualization of microorganisms require a microscope. Microscopes employ lenses that magnify
the original image. A microscope can either be light microscope or electron microscope. Light
microscopes are used to look at intact cells at low magnifications while an electron microscope
looks at internal cell structure and the details of cell surfaces at very high magnification.
The Compound Light Microscope
The common light microscope used in the laboratory is called a compound microscope because
it contains two types of lenses that function to magnify an object. The light microscope uses
visible light to illuminate cell structures.
Types of light microscopes include;
1. Bright-field
2. Phase-contrast
3. Dark-field
4. Fluorescence
The Bright-field microscope is commonly used in laboratory. it uses light rays to produce a dark
image against a bright background. It is used to view fixed and live specimens that have been
stained with basic stains which gives a contrast between the image and the image background.
The Phase-Contrast microscope is commonly used in research because it allows the observation
of wet-mount (living) preparations. It is used to enhance the contrast of images of transparent
and colourless specimens.
The Dark field microscope is a light microscope in which the light reaches the sides only. The
only light reaching the lens is scattered by the specimen and thus the specimen appears light on a
dark background. Dark field microscopy is also an excellent way to observe the motility of
microorganisms.
The fluorescence microscope is used to visualize specimens that emit light of one colour when
light of another colour shines upon them. Cells emit light or fluorescence either because they
contain naturally fluorescent substances such as chlorophyll or other fluorescing components or
because the cells have been stained with a fluorescent dye.

Electron Microscopy
Electron microscopy uses a flow of electron instead of light to produce an image. It has a far
greater
Resolving power than light microscope. This microscope enhances the images of viruses, protein,
lipids, ribosomes, and even small molecules. have far greater resolving power than light
microscope. Two major types of electron microscopy are performed.
Transmission electron microscopy for observing internal cell structure while scanning
electron
microscopy is useful for three-dimensional imaging and for examining surfaces.

Parts of Microscope
1. Arm
It is in the back of the microscope and supports the objectives and ocular. Also, it is the part that we
use to carry or lift it.

2. Base
It’s the bottom of the scope. In addition, it houses the light source and the back section of base acts
as a handle to carry the scope.

3. Course Focusing Knob

We use it to adjust the position of objective lenses. Also, this should be done keeping in mind that
the objective should not hit the slide. In addition, it should be stopped when the object is completely
visible through the ocular.
4. Fine Focusing Knob
We use it to bring the specimen in perfect focus once the specimen is visible through the course-
focusing knob. Also, focus slowly to avoid contact between the objective and the specimen.

5. Illuminator
It is the light source of the microscope.

6. Numerical Aperture or Objective lens

It is found in a compound scope and is the lens that is closest to the specimen.

7. Ocular Lens
This is the lens closest to the viewer in a compound light microscope.

8. Oil immersion Lens

This is a 100x (100 times) objective lens. Also, this lens is small in order to attain high resolution
and magnification. Furthermore, due to its size, it is important for the lens to get as much light as
possible.

Moreover, by immersion of lens in oil it eliminates the refraction of light, it happens because the
glass and oil have almost the same refractive index. Most noteworthy, in this way the light is
maximized and gives the clearest image. Besides, it oils immersion lens is used without oil then the
produced Image will become unclear and has a poor resolution.

The ability to see clearly two items as separate objects under the microscope is called
the resolution of the microscope. The resolution is determined in part by the wavelength of the
light used for observing. Visible light has a wavelength of about 550 nm, while ultraviolet light
has a wavelength of about 400 nm or less. The resolution of a microscope increases as the
wavelength decreases, so ultraviolet light allows one to detect objects not seen with visible light.
The resolving power of a lens refers to the size of the smallest object that can be seen with that
lens. The resolving power is based on the wavelength of the light used and the numerical
aperture of the lens. The numerical aperture (NA) refers to the widest cone of light that can
enter the lens; the NA is engraved on the side of the objective lens.
Figure 1

Light microscopy. (a) The important parts of a common light microscope. (b) How immersion oil
gathers more light for use in the microscope.