INTRODUCTION OF

MICROBIOLOGY
Laboratory of Microbiology
Faculty of Medicine
Brawijaya University
 LABORATORY OF MICROBIOLOGY
TEACHING STAFFS
1. Prof .DR. dr. NOORHAMDANI AS., DMM, SpMK. Head of
Microbiology Laboratory
2. dr. ROEKISTININGSIH, DMM, MS, SpMK.
3. Prof. DR. dr. SANARTO SANTOSO, DTM&H, SpMK. Head of
Instatalation Microbiology Laboratory of Saiful Anwar Hospital
4. Prof. DR. dr. SUMARNO, DMM, SpMK. Head of Study Program
Magister Biomedic
5. DR.Dra. SRI WINARSIH, Apt, MSi. Head of Study Program
Pharmacy
6. dr. DEWI SANTOSANINGSIH, MKes. Head of Infection Control in
Saiful Anwar Hospital.
7. DR. dr .DWI YUNI NUR HIDAYATI, Mkes Person in Charge
Medical Education Microbiology Laboratory Medical Faculty
8. dr . YUANITA MULYASTUTI
9. dr. DEWI ERIKAWATI
10.dr. SIWIPENI IRMAWATI R. M Biomed
11. dr . ANDREW WILLIAM TULLE
TECHNICIANS
1. SLAMET RIYANTO 3. NY.SOEYATI POEJIANI (UCI)
2. WARDI 4. ALI SABET
ADMINISTRATION STAFF
microbiology
A branch of biology which is dealing
with microorganisms and their effects on
other living organisms
Branch of Microbiology :
- Environmental Microbiology
- Space Microbiology
- Marine Microbiology
- Agricultural Microbiology
- Food Microbiology . etc.
- Medical Microbiology
MEDICAL MICROBIOLOGY
Basic Microbiology :
Bacteriology
Virology 22 days ( 16 February 27 March
2015 )
Mycology
TEACHING-LEARNING PROCESS
IN BLOCK BASIC MEDICAL SCIENCE
(BMS
4A)
Lecture FOR SEMESTER II
Meeting
Practical Work
Evaluation
Lecture Meeting :
18 times lecture meeting
Each lecture meeting will be held for 90
minutes

Practical Work
9 times practical work
Each practical work will be held for 180 minutes
Every student has to wear laboratory coat
Every student has to do and assign their
practicum report
Every student has to do staining
 EVALUATION
Mid Semester assesment : 1 Point (A)
Final Semester assessment : 2Point (B)
Practical work assesment : 1Point (C)

total score : A+B+C

4
SCORE
>80 :A
76 80 : B+
70 75 :B
60 69 : C+
56 59 :C
50 55 : D+
45 49 :D
44 :E
 DISCIPLINE

1. LECTURE MEETING PARALEL CLASSES 2 (regular

class)
1
(english class)
1.1 COMPULSARY ATTENDANCE
1.2 PUNCTUAL
1.3 MAX ABSENT 20 %
1.4 PROPER WEAR
1.4 HANDPHONE SILENCED
1.5 ONE RECENT PHOTO OF 4 X 6 (coloured)
(TEST RESULT WILL NOT BE ANNOUNCED IF PHOTO
IS NOT SUBMITTED)
 REFERENCES
1. BAKTERIOLOGI MEDIK (Tim Mikrobiologi FK
UNIBRAW)

2. MEDICAL MICROBIOLOGY (Jawetz,Melnic & Adelbergs)

3. ZINSSER MICROBIOLOGY ( Joklik et al )

4. MICROBIOLOGY, AN INTRODUCTION (Tortora et al )

First Session
A BRIEF HISTORY OF
MICROBIOLOGY

MICROSCOPE
A BRIEF HISTORY OF
MICROBIOLOGY
THE FIRST OBSERVATION

THE DEBATE OVER SPONTANEOUS

GENERATION

THE GOLDEN AGE OF MICROBIOLOGY

THE BIRTH OF MODERN CHEMOTHERAPY :

DREAMS OF A MAGIC BULLET

MODERN DEVELOPMENTS IN MICROBIOLOGY

 THE FIRST OBSERVATION
1665 An Englishman, Robert Hooke,
reported to the world that lifes smallest structural units were
little boxes or cells
Using his improving version of a compound microscope
he was able to see individual cells beginning of the cell theory
that all living things are composed of cells

1673 1723, Antony van Leeuwenhoek

The Dutch merchant and amateur scientist was the first to actually
observe live microorganism through his simple single lens microscope
the animalcules in rainwater, in liquid in which peppercorn had
soaked, and in material scraped from his teeth
He drew the basic form of bacteria as coccus, rods and spiral
coccus

spiral

rods
THE DEBATE OVER SPONTANEOUS GENERATION
Until the second half of the 19th century many scientists and philosophers
(Aristoteles, Samson, Virgil) believed that some form of life could arise
spontaneously from nonliving matter spontaneous generation

People commonly believed that toads, snakes, and mice could be born
of moist soil; that flies could emerge from manure; and that maggots
could arise from decaying corpses

Evidence Pro and Con

1668 Fransesco Redi, the Italian physician, a strong opponent of
spontaneous generation demonstrate that maggots do not arise
spontaneously from decaying meat
Redi filled three jars with decaying meat and sealed them tightly.
Then he arranged three other jars similarly but left them open. Maggots
appeared in the open jars.Sealed containers showed no signs of maggots

Redis antagonists were not convinced; they claimed that fresh air was
needed for spontaneous generation
THE DEBATE OVER SPONTANEOUS GENERATION
Redi set up a second experiment three jars were covered with a fine
net instead of being sealed. No larvae appeared in the gauze-covered jars

Many scientists still believed that small organisms (van Leeuwenhoeks

animalcules) were simple enough to be generated from nonliving materials

1745 : spontaneous generation seem to be strengthened, when

John Needham, an Englishman, found that even after he heated nutrient
fluid before pouring them into covered flasks, the cooled solution were
soon teeming with microorganisms.
Needham claimed that microbes developed spontaneously from the fluid

Twenty years later : Lazzaro Spallanzani, an Italian scientist, suggested

that microorganisms from the air probably had entered Needhams
solutions after they were boiled. Spallanzani showed that nutrient fluids
heated after being sealed in a flask did not develop microbial growth

Needham responded by claiming the vital force necessary for sponta-

neous generation had been destroyed by the heat and was kept out of
the flasks by the sealed
THE DEBATE OVER SPONTANEOUS GENERATION

Laurant Lavoisier showed the importance of oxygen to life.

Spallanzanis observations were criticized on the grounds that there was

not enough oxygen in the sealed flasks to support microbial life

The Theory of Biogenesis

Rudolf Virchow challenged spontaneous generation with the concept

of biogenesis, he claim that living cells can arise only from preexisting
living cells

Arguments about spontaneous generation continued until 1861, when the

issue was resolved by the French scientist Louis Pasteur

With a series of ingenious experiments, Pasteur demonstrate that

microorganisms are present in the air and can contaminate sterile
solutions, but air itself does not create microbes
 Pasteurs experiments disproving the theory of
spontaneous generation

(1) Pasteur first poured beef broth into a long-necked flask. (2) Next he heated the neck of the flask
and bent it into an S-shaped curve; then he boiled the broth for several minutes. (3)
Microorganisms did not appear in the cooled solution, even after long periods
 Pasteur showed that microorganisms can be present in
nonliving matter on solids, in liquids, and in the air

He demonstrated that microbial life can be destroyed

by heat form the basis of aseptic techniques

The debate of Spontaneous Generation disproved

However still have a problem of spores resistant to

heat John Tyndall (1820-1893) TYNDALLIZATION
Spontaneous Generation Theory totally finished
The Golden Age Of Microbiology
1857 - 1914
Pasteur & Robert Koch, led to the
establishment
of microbiology as a science
Discoveries during these years included
both the agents of many diseases and the
role of immunity in the prevention and
cure of disease
Fermentation and Pasteurization
The Germ Theory of Disease
Vaccination
 Fermentation & Pasteurization
A group of French merchants asked Pasteur to find out why
wine and beer soured a method that would prevent
spoilage ?
Many scientist believed that air converted the sugars in
these fluids into alcohol
Pasteur found instead that microorganisms called yeasts
convert the sugars to alcohol in the absence of air. This
process, called fermentation, is used to make wine and
beer
Souring and spoilage are caused by different micro-
organisms called bacteria. In the presence of air, bacteria
change the alcohol in the beverage into vinegar (acetic acid)
 The Germ Theory of Disease
Before the time of Pasteur, effective treatments for
many diseases were discovered by trial and error,
but the causes of the disease were unknown
The realization that yeasts play a crucial role in
fermentation was the first link between the activity of
a microorganism and physical and chemical changes
in organic materials
This discovery alerted scientists to the possibility
that microorganisms might have similar relationships
with plants animals - specifically, that
microorganisms might cause disease. This idea was
known as the germ theory of disease
 The Germ Theory of Disease
1840 Ignaz Semmelweis a Hungarian physician, had
demonstrated that physician, who at the time did not
disinfect their hands, routinely transmitted infections
(puerperal, or childbirth, fever) from one obstetrical
patient to another
1860 Joseph Lister an English surgeon, applied the
germ theory to medical procedure. Disinfectants were not
used at the time, but Lister knew that phenol (carbolic
acid) kills bacteria, so he began treating surgical wounds
with a phenol solution
1876 Robert Koch, a German physician, proved that
bacteria actually cause disease discovered rod-shaped
bacteria now known as Bacillus anthracis in the blood of
cattle that had died of anthrax
 The Germ Theory of Disease
Kochs research provides a framework for the study of the
etiology of any infectious disease Koch Postulates :
1. The same pathogen must be present in every case of
disease
2. The pathogen must be isolated from the diseased host
and grown in pure culture
3. The pathogen from the pure culture must cause the
disease when it is inoculated into a healthy,
susceptible
laboratory animal
4. The pathogen must be isolated from the inoculated
animal and must be shown to be the original organism
 VACCINATION
1796 Edward Jenner, a young British physician,
embarked on an experiment to find a way to protect
people from smallpox :
- A young milkmaid informed Jenner that she couldnt get
smallpox because she already had been sick from
cowpox
- First Jenner collected scrapings from cowpox blisters.
Then
he inoculated a healthy 8-year-old volunteer with the
cowpox material by scratching the persons arm with a
pox-contaminated needle. In a few days, the volunteer
became mildly sick but recovered and never again
contracted either cowpox or smallpox. The process was
called vaccination (vacca=cow)
- The protection from disease provided by vaccination is
called immunity
 VACCINATION
1880 Pasteur discovered why vaccination work. He found
that the bacteria that causes fowl cholera lost its ability
to cause disease after it was grown in the laboratory for
long periods. However it and other microorganisms with
decreased virulence was able to induce immunity against
subsequent infections by its virulent counterpart.
Some vaccines are still produced from avirulent microbial
strains that stimulate immunity to the related virulent strain.
Other vaccines are made from killed virulence microbes,
from isolated components of virulent micoorganisms, or by
genetic engineering techniques
The Birth of Modern
Chemotherapy :
Dreams of a between
After the relationship Magic Bullet
microorganisms and
disease was established, medical microbiologists next
focused on the search for substances that could destroy
pathogenic microorganisms without damaging the
infected animal or human
Treatment of disease by using chemical substances is
called chemotherapy
Chemotherapeutic agents prepared from chemicals in
the laboratory are called synthetic drugs
Chemicals produced naturally by bacteria or fungi to act
against other microorganisms are called antibiotics
 The First Synthetic Drugs
Paul Ehrlich, a German physician, was the imaginative
thinker who fired the first shot in the chemotherapy
revolution. Ehrlich speculated about a magic bullet
that could destroy pathogen without harming the
infected host
In 1910, he found a chemotherapeutic agent called
salvarsan, an arsenic derivative effective against
syphylis
By the late 1930s, researchers had developed several
other synthetic drugs, mostly were derivatives of dyes
that could destroy microorganisms
In addition, Domagk (1935) discovered that prontosil
had dramatically effect against streptococcal infections
in the body was changed into sulfanilamide that
analog with PABA
 A Fortunate Accident - Antibiotics
The first antibiotic was discovered by accident
1928 Alexander Fleming, a Scottish physician and
bacteriologist, looked at the curious pattern of growth on the
contaminated plates. There was a clear area around the mold
where the bacterial culture had stopped growing
The mold was later identified as Penicillium notatum, and
Fleming named the molds active inhibitor as penicillin.
The enormous usefulness of penicillin was not apparent until
the 1940s Florey & Chain
1939 Rene Dubos, a French microbiologist, discovered
two antibiotics called gramicidin and tyrocidine. Both were
produced by a bacterium, Bacillus brevis, cultured from soil.
Modern Development in
Microbiology
Bacteriology

Mycology

Parasitology

Immunology

Virology

Recombinant DNA technology

 VIROLOGY
The study of virus, actually originated during the Golden Age
of Microbiology
1892 Dmitri Iwanowski reported that the organism that
cause mosaic disease of tobacco was so small that it passed
through filter fine enough to stop all known bacteria
1935 Wendell Stanley demonstrated that the organism,
called tobacco mosaic virus (TMV), was fundamentally
different from other microbes and so simple and
homogeneous that it could be cristallized like chemical
compound
1940 Since the development of the electron microscope,
microbiologists have been able to observe the structure of
viruses in detail.
 Recombinant DNA Technology
Until the 1930s, all genetic research was based on the study of
plant and animal cells
In the 1940s, the scientists turned to unicellular organisms,
primarily bacteria
In 1941 George W.Beadle and Edward L.Tatum
demonstrated the relationship between genes and enzymes
1944 Oswald Avery, Colin McLeod, and Maclyn McCarty
DNA was established as the hereditary material
1946 Joshua Lederberg and Edward L.Tatum discovered that
genetic material could be transferred from one bacterium to
another by a process called conjugation
1953 James Watson and Francis Crick proposed a model for
the structure and replication of DNA
MICROSCOPE
MICROSCOPE
Compound Light Microscopy
Darkfield Microscopy
Phase-contrast Microscopy
Fluorescence Microscopy
Confocal Microscopy
Electron Microscopy
Compound Light Microscope
Has a series of lenses and use visible light as its
source of illumination
A series of finely ground lenses forms a clearly
focused image that is many time larger than the
specimens itself
This magnification is achieved when light rays
from an illuminator condensor specimen
objective lenses ocular lens
Total magnification = objective lens magnification
x ocular lens magnification
Objective lens : 10 x (low power), 40 x (high
power), and 100 x (oil immersion)
Ocular lens : 10 x
Compound Light Microscope
The total magnifications would be 100 x for low
power, 400 x for high power , and 1000 x for oil
immersion. Some compound light microscopes
can achieve a magnification of 2000 x with the
oil immersion lens.
The oil immersion has the same refractive
index as glass, so the oil becomes part of the
optics of the glass of the microscope.
Unless immersion oil is used, light rays are
refracted as they enter the air from the slide
Compound Light Microscope
 Darkfield Microscopy
Is used for examining live microorganisms that
either are invisible in the ordinary light microscope,
cannot be stained by standard methods, or are so
distorted by staining that their characteristics then
cannot be identified
A darkfield microscope uses a darkfield condensor
that contain an opaque disc the specimen
appears light against a black background
One use for darkfield microscopy is the examination
of very thin spirochetes, such as Treponema
pallidum, the causative agent of syphilis
 Phase-Contrast Microscopy
Is especially useful because it permits detailed examination
of internal structures in living microorganisms
The principle of phase-contrast microscopy is based on the
wave nature of light rays, and the fact that light rays can be
in phase (their peaks and valleys match) or out of phase.
In phase-contrast microscopy, one set of light rays comes
directly from the light source. The other set comes from light
that is reflected or diffracted from particular structure in the
specimen
Phase-contrast microscope is provided with diffraction plate
Brightfield Darkfield Phase-contrast
 Fluorescence Microscopy
The object is stained with one of a group of fluorescent
dyes called fluorochromes (primolin, acridine orange
R,
thiazo yellow-G, auramine O, fluorescein
isothiocyanate)
The light source Ultraviolet light
The principal use of fluorescence microscopy is a
diagnostic technique called fluorescence-antibody
(FA) technique, or immunofluorescence
Confocal Microscopy
A fairly recent development in light microscopy
Like fuorescent microscopy, specimens are
stained with fluorochromes
The light source laser
Most confocal microscopes are used in
conjunction with computers to construct three-
dimensional images
Can be used to evaluate cellular physiology by
monitoring the distributions and concentrations
of substances such as ATP and calcium ions.
Electron Microscopy
Object smaller than about 0.2 m, such as
virus, or the internal structures of cels must be
examined with an electron microscope
A beam of electrons is used instead of light
Objects are generally magnified 10,000
100,000 x
Instead of using glass lenses, an electron
microscope uses electromagnetic lenses to
focus a beam of electrons onto specimen
There are two types of electron microscope :
- The transmission electron microscope
- The scanning electron microscope