Department of Medical Laboratory, HHSC,

• Module name: Medical Microbiology

• Course: Medical Bacteriology and food MB
• Course code: MeLs
• Credit hours/ECTS: 9
• Category of student: MeLS; Year-III; Semester-I
• Instructor: Wondimagegn
 Course Content

1. Introduction to Microbiology

2. Bacterial cells

3. Bacterial taxonomy

4. Nutrition and metabolism of bacteria

8. Bacterial genetics
 Course content

9. Disinfection and sterilization

10. Principles of staining

11. Preparation of culture media and isolation of bacteria

12. Biochemical and serological tests

13. Antimicrobial chemotherapy and susceptibility testing

 1. Introduction to Microbiology
1.1. Definition: Microbiology is a science defined by smallness.
• Its creation was made possible by the invention of the microscope
(Gr. micro, small skop, to look, see)

• The microscope allowed visualization of structures too small

to see with the naked eye

• The major classes of microorganisms include bacteria, fungi,

parasites and the non living virus particles
 1. Introduction…
• Microbiology considers the microscopic forms of life and deals
about
• Reproduction and physiology
• participation in the process of nature
• helpful and harmful relationship with other living things
• significance in science and industry
 1. Introduction…
1.2. Branches of microbiology
• Food microbiology: study microorganisms (MOs) causing
food spoilage and food borne illness
• Agricultural microbiology: study agriculturally relevant
MOs
• Veterinary microbiology: study the role of microbes in
veterinary medicine
• Environmental microbiology: study the function and
diversity of microbes in their natural environments
 1. Introduction…
• Medical microbiology: study pathogenic microbes and also
– Study microbial pathogenesis
– Study epidemiology of microbial diseases
– Related to the study of immunology
– Study their diagnosis
– Study the prevention and control
• Mainly comprise: Bacteriology, Virology, Mycology
 1. Introduction…
• Microbes have both advantages and disadvantages
– In the ecology: Microbes
 are producers: provide energy to ecosystems
 are fixers: make nutrients available from inorganic sources,
e.g., nitrogen
 are decomposers: free up nutrients from dead sources
 form symbioses (fungi associated with plant roots, the bacteria
found in legume root nodules, etc.)
 serve as endosymbionts (e.g., chloroplasts and mitochondria)
 1. Introduction…
– In the industry:
 Fermentation products (ethanol, acetone, etc.)

 Food and drinks Wine, cheese, yogurt, bread, half-sour

pickles, etc.
 Biotech: Recombinant products (e.g., human insulin, vaccines)

 Environment: Bioremediation
 1. Introduction…
– Microbes and Disease
 Microbes both cause and prevent diseases

 Microbes produce antibiotics used to treat diseases

 The single most important achievement of modern medicine is the

ability to treat or prevent microbial disease
Knowledge of Microbes allows humans to
• Prevent food spoilage
• Prevent disease occurrence
• Led to aseptic techniques to prevent contamination in
medicine and in microbiology laboratories.
 2. History of Microbiology
• The history of microbiology is related to how people try to
define disease at different times in history
• Ancient times: disease was believed to be punishment of God
• Many philosophers during the early period in history, however,
aware of the belief that disease was transmitted by invisible
“animals” since the animal could not be seen, the theory
remains just that a theory.
 2. History of Microbiology…

• Hippocrates: ill health resulted due to changes in air, winds,

water, climate, food, nature of soil and habits of people.
• Varro: Disease was caused by animate particles invisible to
naked eye where carried in air via the mouth and nose to body.
• Fracastorius: Agent of communicable disease was living
germs that transmitted by direct contact and indirect contact
with human, animal and objects respectively.
• No proof b/c no experimental study
2. History of Microbiology…
• The first microbes were observed in 1673.

• In 1665, Robert Hooke (Englishman) reported that living

things were composed of little boxes or cells.
2. History of Microbiology…
• Antony van Leeuwenhoek (1632-1723 G.C): Dutch scientist

– Observe microscopic organisms in pond water and debris

surrounding the teeth
– Leeuwenhoek called this organism “animalcules” meaning
little animals from sample taken on human mouth.
– Later named Bacteria

– He was the first to describe different shapes of bacteria as

rod, spherical and spiral.
 2. History of Microbiology…

• Question: where did these organisms originated from?

• Many scientists were searching for an explanation for

spontaneous appearance of living things from decaying material,
stagnant ponds, infected wound, fermented grains…

• On the bases of this observation, two theories were formulated.

– Theory of Abiogenesis

– Theory of Biogenesis
2. History of Microbiology…
1. Abiogenesis (spontaneous generation): states that living things
originated “spontaneously” from non-living things.
• Aristotle (384-322 BC): The founder of spontaneous
generation.
– He observed spontaneous existence of fishes from dried
ponds, when the pond was filled with rain.
 2. History of Microbiology…
• Spontaneous history myths
 Maggots from rotting meat

 Fleas from hair

 Flies from fresh and rotting fruit

 Mosquitoes from stagnant pond water

 Locusts from green leaves

 Termites are generated from rotting wood

2. History of Microbiology…
2. Biogenesis: States that life comes from pre existing life
a. Francesco Redi (1626-1697): He suggested that flies land on
pieces of exposed meat and lay their eggs, which then hatch to
maggots.
• Utilized jars containing meat. Some were covered, some
were not.
• Maggots appeared in uncovered jars.
• Results not accepted for microscopic organisms.
Jar-1 Jar-2
Jar-3
Left open Covered with netting Sealed
Maggots appeared on No maggots developed
Maggots developed
the netting
Flies were observed
Flies were observed
laying eggs on the
laying eggs on the
meat in the open
netting
jar
2. History of Microbiology…

b. John Needham (1749)

• Performed experiments similar to Redi’s on the origin of life
in microscopic organisms
• Introduced the first culture medium for microbial growth.
• Utilized infusion broth prepared by boiling meat, grain, etc.
• Turbidity indicated growth.
• Broth put in flasks, some were sealed with corks, and some
were not.
 2. History of Microbiology…

• All flasks became cloudy, result different from Redi’s

experiment.
• He suggested that life originate spontaneously from
nonliving matters
• The spontaneous generation opponents didn’t accept his
conclusion, they said it could be due to entrance organisms
from air, flasks or improper seal
• Hard to kill endospores—boiled broths not always sterilized
2. History of Microbiology…

c. Lazzaro Spallanzani (1776)

• Repeated Needham’s experiments to disproof spontaneous
generation in microscopic life.
• Boiled broth after placing in flasks.
• Sealed flasks by plugging with solid stopper.
– Boiled and sealed, did not turn cloudy, microbes not found
– Occasionally sealed flask  cloudy.
2. History of Microbiology…

• Not accepted by spontaneous generation supporters, because:

boiling altered broths so as to prevent spontaneous

generation
absence of air prevented spontaneous generation

heating or chemically treating air removed vital force from

air thereby preventing spontaneous generation
 2. History of Microbiology…

d. Louis pasture (1822- 1895)

• Disproved the theory of abiogenesis once and for all.
• He placed broth in a large curved flask/swan-necked (pasture
goose neck flask)
• Moisture were trapped at the bend and no growth was observed
(Fig A)
• He tilted the flask and broth contacted with dust particles
trapped at the bend and growth was observed (Fig B)
Air freely moved through the tube, but dust particles were
trapped in the curved portion of the flask and no microbial
growth was observed.
Therefore, Pasteur proved that microorganisms entered to the
broth with the air and micro organisms did not evolve
spontaneously
 2. History of Microbiology…
• Other major contributions of Louis Pasteur
– Microbial theory of fermentation
– Principles and practices of sterilization and pasteurization
– Control of disease of silk worm
– Development of vaccines against anthrax and rabies
– Discovery of streptococci.
 2. History of Microbiology…

• Germ theory of disease is a theory that tries to explain the

association between disease and MOs
• Pasture has developed the germ theory of diseases, which states
that a specific disease is caused by a specific type of MOs.
– But this was not experimentally demonstrated

• Robert Koch, in 1876 established an experimental procedure to

prove the germ theory of disease and his scientific procedure is
known as Koch’s Postulate.
 2. History of Microbiology…

• He demonstrated the theory by studying anthrax, a disease of

cows
– He isolated the organism in a pure culture
– He injected the isolate to a healthy animal
– The injected animal became ill and developed anthrax
– The causative agent was re-isolated from the infected
animal
 2. History of Microbiology…

• This sequence of isolation, re-infection and recovery of the

infectious agent is called Koch’s postulate.
• Based on his observation Koch concluded the following
• A Micro-organism can be accepted as a causative agent of an
infectious disease only if the following conditions are
satisfied.
1. The microorganism should be found in every case of the
disease and under conditions, which explain the pathological
 4. The Germ Theory of Disease …

2. It should be possible to isolate the causative agent in pure

culture from the lesion
3. When such pure culture is inoculated in to appropriate lab
animal, the lesion of the disease should be reproduced.
4. It should be possible to re-isolate the causative agent in pure
culture from the lesion produced in the experimental animal.
 2. History of Microbiology…

• Exceptions to Koch’s postulates

1. Many healthy people carry pathogens but do not exhibit

symptoms of the disease.
2. Some microbes are very difficult or impossible to grow
in vitro (in the laboratory) in artificial media.
3. Many pathogens are species specific.
4. Certain diseases develop only when an opportunistic
pathogen invades imuno-compromised host.
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