Microbiology veteriner1 pendahuluan

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MICROBIOLOGY
VETERINER
PROGRAM KEDOKTERAN HEWAN
PROGRAM STUDI PENDIDIKAN DOKTER HEWAN
Dr.Sri Murwani,drh,MP

09/21/13www.themegallery.com Company Logo
Dr.Sri Murwani, drh,MP
drh. Masdiana C. Padaga, M.AppSc
Program Kedokteran Hewan-UB, Program Studi Pendidikan Dokter Hewan
drh. Aldilla Noviantri
drh. Nurina Titisari

3rd
SEMESTER 4th
SEMESTER
MICROBIAL BIOLOGY
BAKTERIOLOGY
VIROLOGY
MIKOLOGY
3 credit points
IMMUNOLOGY
3 credit points
MICROBIAL
INFECTION
3 credit points

Lecture meeting
Structured Program
(Seminar)
Laboratory Practical
Evaluation:
Middle assessment (25%
Final assessment (25%)
Presentation (15%)
Quiz (10%)
Lab.pract. Assessment
(25%)

COMPULSARY
ATTENDANCE
DICIPLINE
punctual
attendance > 80 %
proper wear
Hp silenced
1 colour recent
Photo 3x4

A BRIEF HISTORY
OF MICROBIOLOGY

Microbiology ????? Microorganisms
Bacteria
Virus
Fungi
Parasite ?? --- protozoa
Microscopic algae

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The branch of
biology that
deals with
microorganisms
and their effects
on other living
organisms
Microbiology
What the m.o function ?
Balancing
-Soil microbes
-Photosynthetic m.o
-Intestine bacteria
-Commercial applications
-Etc.

Pathogenic m.o
Non-Pathogenic m.o
Opportunistic m.o
Microorganisms
Normal Flora m.o

Many branches of Microbiology :
- Environmental Microbiology
- Space Microbiology
- Marine Microbiology
- Agricultural Microbiology
- Food Microbiology … etc.
- Medical Microbiology
Vet. Microbiology

Contents
MODERN DEVELOPMENTS IN MICROBIOLOGY
THE BIRTH OF MODERN CHEMOTHERAPY :
DREAMS OF A “MAGIC BULLET”
THE GOLDEN AGE OF MICROBIOLOGY
THE DEBATE OVER SPONTANEOUS
GENERATION
THE FIRST OBSERVATION

THE FIRST OBSERVATION
1665 – An Englishman, Robert Hooke,
reported to the world that life’s 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 of soaked peppercorn, and in
material scraped from his teeth
the basic form of bacteria as coccus, rods and
spiral

coccus
rods
spiral

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 atau abiogenesis
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
cover tightly open
no maggots maggots
Redi’s antagonists were not convinced; they claimed that
fresh air was needed for 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
Leeuwenhoek’s 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 Needham’s 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 spontaneous generation had been destroyed
by the heat and was kept out of the flasks by the sealed
Laurant Lavoisier showed the importance of oxygen to life
Spallanzani’s 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

Pasteur’s 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
 Chemical activities m.o
 Improved the techniques for performing microscopy &
culturing mo
 Vaccination
 Surgical techniques

Fermentation & Pasteurization
A group of French merchants asked Pasteur to find out why
wine and beer soured  a method that would prevent
spoiled ?
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 spoiled 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

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

Koch’s 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 disease 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 couldn’t get
smallpox because she already had been sick from cowpox
- Jenner scrapings from cowpox blisters inoculated to
a healthy 8-year-old volunteer by scratching the 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 “ Magic Bullet “
After the relationship between 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 mold’s 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.

Mycology
Eukaryotic
Morphologically divided into 2 different types:
1. Filamentous fungi
2. Yeast
Mostly are opportunistic pathogen, rare cause serious
disease in healthy people

Recombinant DNA Technology
1928 by Griffith study about pathogenecity of Diplococcus
pneumoniae. The important material of this bacteria is it
capsule which can be move between bacteria.
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
1946 – Joshua Lederberg and Edward L.Tatum
discovered that genetic material could be transferred from
one bacterium to another by a process called conjugation
Genetic engineering: induction hormone production,
vaccines, diagnostic tool, etc …..

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

 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

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 fLuorescent 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 µm, 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

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