INTRODUCTION TO MICROBIOLOGY matter.

They also decompose dead plants

and animals.
 Microbiology is the study of living organisms
that are too small to be seen by the naked 4. Agriculture – farming practices have been
eye. It involves the biological activities of improved by recognizing and capitalizing on
microorganisms such as bacteria, yeasts, the role of microorganisms. Microbes have
molds, protozoa, viruses, and rickettsia. been used to decompose crop residues and
Microorganisms are otherwise called organic matter for fertilizers. Nitrogen-fixing
microbes or “germs”. microbes have been helpful in the growth of
leguminous crops, some microbes are also
The study of microorganisms includes the symbionts to ruminant animals. Recently,
following form, structure, reproduction, some microbes have been used for the
metabolism, and identification. It also includes: biological control of pests and diseases in
plants and for genetic engineering.
1. Study their distribution in nature.
2. Relationship to each other and other 5. In research – microbes are extensively used
living things. to provide explanations on:
3. Beneficial and detrimental effects to 1. Growth and population phenomena
man. 2. Cell division, metabolic pathways
4. Physical and chemical changes they 3. Molecular genetics
cause within the environment. 4. Absorption and permeability of cells

Microorganisms occur everywhere in nature. A Brief History of Microbiology

Since the conditions that favor the survival and Microbiology has had a long, rich history, initially
growth of microbes are those under which centered in the causes of infectious diseases
people normally live, it is inevitable that we live but now including practical applications of the
among the multitudes of microbes. Microbes science. Many individuals have made
are found everywhere except in: significant contributions to the development of
1. Blood and internal tissues of animals. microbiology.
2. Internal tissues of normal plants.
Early history of microbiology
3. Tip of active volcano.
Historians are unsure who made the first
observations of microorganisms, but the
Important roles of Microorganisms
microscope was available during the mid‐1600s,
1. Medicine – world health has been improved
and an English scientist named Robert
through the discovery of the microbial
Hooke made key observations. He is reputed to
causes of most human diseases, leading to
have observed strands of fungi among the
the development of vaccines, antibiotics,
specimens of cells he viewed. In the 1670s and
and chemical agents to combat these
the decades thereafter, a Dutch merchant
diseases.
named Anton van Leeuwenhoek made careful
2. Food – foods have been improved in quality observations of microscopic organisms, which
and protected from spoilage to enable he called animalcules. Until his death in 1723,
wide distribution and storage. van Leeuwenhoek revealed the microscopic
world to scientists of the day and is regarded as
3. Environment – methods have been
one of the first to provide accurate descriptions
developed to break the chain of diseases
of protozoa, fungi, and bacteria.
transfer through waterborne diseases.
Microbes also enhance the quality of rivers After van Leeuwenhoek died, the study of
and lakes by degrading naturally organic microbiology did not develop rapidly because
microscopes were rare and the interest in
microorganisms was not high. In those years, set of principles whereby other microorganisms
scientists debated the theory of spontaneous could be related to other diseases.
generation, which stated that microorganisms
arise from lifeless matter such as beef broth. This
theory was disputed by Francesco Redi, who
showed that fly maggots do not arise from
decaying meat (as others believed) if the meat
is covered to prevent the entry of flies. An
English cleric named John Needham advanced
spontaneous generation, but Lazzaro
Spallanzani disputed the theory by showing that
boiled broth would not give rise to microscopic
forms of life.

Louis Paster and the germ theory. Louis

Pasteur worked in the middle and late 1800s. He
performed numerous experiments to discover
why wine and dairy products became sour, and
he found that bacteria were to blame. Pasteur
called attention to the importance of The steps of Koch's postulates used to relate a
microorganisms in everyday life and stirred specific microorganism to a specific disease. (a)
scientists to think that if bacteria could make Microorganisms are observed in a sick animal
the wine “sick,” then perhaps they could cause and (b) cultivated in the lab. (c) The organisms
human illness. are injected into a healthy animal, and (d) the
animal develops the disease. (e) The organisms
Pasteur had to disprove spontaneous are observed in the sick animal and (f)
generation to sustain his theory, and he reisolated in the lab.
therefore devised a series of swan‐necked
flasks filled with broth. He left the flasks of broth The development of microbiology
open to the air, but the flasks had a curve in the In the late 1800s and for the first decade of the
neck so that microorganisms would fall into the 1900s, scientists seized the opportunity to further
neck, not the broth. The flasks did not become develop the germ theory of disease as
contaminated (as he predicted they would enunciated by Pasteur and proved by Koch.
not), and Pasteur's experiments put to rest the There emerged a Golden Age of
notion of spontaneous generation. His work also Microbiology during which many agents of
encouraged the belief that microorganisms different infectious diseases were identified.
were in the air and could cause disease. Many of the etiologic agents of microbial
Pasteur postulated the germ theory of disease, disease were discovered during that period,
which states that microorganisms are the leading to the ability to halt epidemics by
causes of infectious disease. interrupting the spread of microorganisms.
Despite the advances in microbiology, it was
Pasteur's attempts to prove the germ theory rarely possible to render life‐saving therapy to
were unsuccessful. However, the German an infected patient. Then, after World War II,
scientist Robert Koch provided the proof by the antibiotics were introduced to medicine.
cultivating anthrax bacteria apart from any The incidence of pneumonia, tuberculosis,
other type of organism. He then injected pure meningitis, syphilis, and many other diseases
cultures of the bacilli into mice and showed that declined with the use of antibiotics.
the bacilli invariably caused anthrax. The
procedures used by Koch came to be known Work with viruses could not be effectively
as Koch's postulates (Figure ). They provided a performed until instruments were developed to
help scientists see these disease agents. In the
1940s, the electron microscope was developed The germ theory of disease states that "every
and perfected. In that decade, cultivation infectious disease has its invisible living cause”;
methods for viruses were also introduced, and this theory was advocated by Louis Pasteur
the knowledge of viruses developed rapidly. (1865) when he studied the silkworm disease
With the development of vaccines in the 1950s that destroyed the silk industry in France.
and 1960s, such viral diseases as polio, measles, Infected silkworm moths could be recognized
mumps, and rubella came under control. microscopically. When females free of infection
were used, the silkworm remained free of
Modern microbiology infection.
Modern microbiology reaches into many fields
Joseph Lister (1827-1912) discovered that
of human endeavor, including the
infections were the result of living microbes. He
development of pharmaceutical products, the
then used diluted carbolic acid in wound
use of quality‐control methods in food and dairy
dressings and during Surgical procedures. He
product production, the control of disease‐
was later called the "Father of antiseptic
causing microorganisms in consumable waters,
techniques”
and the industrial applications of
microorganisms. Microorganisms are used to In 1843-1910, Robert Koch discovered the
produce vitamins, amino acids, enzymes, and anthrax bacillus which he cultured in the
growth supplements. They manufacture many laboratory and then injected them in other
foods, including fermented dairy products (sour healthy animals from which he was able to
cream, yogurt, and buttermilk), as well as other isolate the original bacilli after they contracted
fermented foods such as pickles, sauerkraut, the disease. Thus, the Koch's postulates were
breads, and alcoholic beverages. formulated.

Historical Background Microbiology 1. A microorganism must be present in all

1. Prehistoric Period – period of speculation cases of disease and absent in all
2. Supernatural – advocators of this theory healthy individuals.
attributed diseases as a punishment of 2. The microorganism in a suspected case
wickedness. could be isolated in pure culture.
3. Natural theory – proponents of this theory 3. The isolated organism should be able to
believed that diseases were caused by cause or reproduce the disease in a
comets, eclipses and earthquakes. susceptible animal
4. Miasmatic theory – this theory believes that 4. The organism can be re-isolated or
diseases were caused by particular vapors in recovered from an experimentally
the air. infected animal
5. Tb, Diptheria, typhoid fever, cholera and
The Early Development of Microbiology gonorrhea.
Microorganisms were discovered with the
discovery of the microscope by Antoine van Robert Koch is also called the "Father of
Leeuwenhoek (1632-1723). A Dutch merchant Bacteriologic Technique" because he devised a
who grounded lenses and made a simple method of growing or culturing bacteria on
microscope with a magnification of 300x. With agar media. He also discovered the Koch's
the simple microscope, he observed organisms bacillus or Mycobacterium tuberculosis.
in rainwater, seawater, and teeth scrapings In 1798, Edward Jenner presented a paper and
which he called "animalcules”. These were later pointed out that many of the milkmaids in
identified to be protozoans and bacteria. England were immune to smallpox. He stated
that their immunity stemmed from an infection
Germ theory of Disease "Von Plenciz" with cowpox virus, a disease not uncommon in
cows. He then invented the vaccine for
smallpox, thus he was called the "Father of Staining Procedures
Immunology”. Staining – process of application of colored
compounds. Dyes or stains on surfaces and
There are many other scientists who earned
render it colored or visible.
recognition in the history of Microbiology. Some
even died after they were infected by the
Purpose of staining
organisms they were studying. G.T. Gaffky
1. To render structures to become visible
isolated the typhoid bacillus; F. A. Loeffler
which are semi-transparent
identified the diphtheria bacillus and S. Kitasato
2. They reveal the shape, forms and sizes
cultured tetanus bacillus. Early in the 20th
3. To show the presence of external and
century, Howard Taylor Ricketts accidentally
internal structures
contracted typhus during the course of his
4. To produce specific chemical and
research and died of it.
physical reaction

Microscopes and Microscopy

Types of Staining Techniques
Microscope – an instrument or device used to
1. Simple Staining technique – one stain eg.
magnify and see specimens that are too small
Methylene blue
and can't be seen by the naked eye.
2. Differential staining technique – shows
difference between two cell types
Two kinds of Microscope
3. Gram Staining technique – by Christian
1. Simple microscope – possess only one lens
Gram divides the bacteria into
2. Compound microscope – has two lenses
4. Gram positive (+) – colored violet; absorbs
a. ocular lens/ eyepiece
crystal violet stain
b. objective lens
5. Gram-negative bacteria (G-) – colored pink,
absorb the counterstain carbol fuchsin or
Two Kinds of Microscope as to source of
safranin
illumination
6. Acid-fast staining technique – helps to
1. Light microscope – natural or artificial light
identify mycobacterium
2. Electron microscope – beams of electrons to
light the specimens
Isolation of bacteria into Pure Culture
3. Scanning electron microscope
In nature, microbes are found in mixtures so
4. Transmission electron microscope
they have to be isolated into pure culture to
have & a true picture of its:
Kinds of Light Microscope
1. appearance, color and structure
1. Bright-Field Microscope – the specimen
2. cultural characteristics
appears dark against a bright background.
3. metabolic activities
2. Dark-field microscope – the specimen
4. importance
appears bright against a dark background.
3. Phase-contrast microscope – Phase contrast
Culture – refers to the cultivation of organisms
objective is installed to increase the contrast
Culture media – a substance where
between the specimen and the
microorganisms are grown in the laboratory
background
Microbial culture – growing of microbes
4. Fluorescence
5. Ultraviolet microscope
Types of Culture
6. X-ray microscope
1. Pure Culture – culture of only one species of
7. Polarization microscope
organisms
2. Mixed culture – growth/culture of two or
Preparations for Light Microscopic Examination
more species
1. Wet mounts
2. Fixed prepared slides
Methods of obtaining Pure Culture
1. Enrichment Procedure
2. Isolation into colonies
3. Streak plate method
4. Pour plate method
5. Spread plate method
6. Serial dilution technique
7. Single cell technique

Kinds of Culture Media

As to appearance or Physical State

1. Liquid – eg. nutrient broth
2. Solid – eg. nutrient agar, agar is the
solidifying agent

As to chemical composition
1. Synthetic culture media – the exact
chemical component are known
2. Non-synthetic culture media – unknown
chemical components eg. blood

As to function
1. General Culture media – for general use,
could be used in all types of microbes. Eg.
nutrient broth and nutrient agar
2. Selective culture media – use for growing
specific microbes
3. Salmonella Shigella agar (S-S agar) - for the
culture of Salmonella that causes typhoid
and Shigella that causes dysentery
4. Differential culture media culture media that
shows differences between two species.
5. Eosin-Methylene Blue Agar produces
chaotic growth of E.coli where colonies
appear as greenish metallic green
while Enterobacter aerogenes appear as
pink colonies.