COLLEGE OF AGRICULTURE AND NATURAL RESOURCES

MANAGEMENT
General Microbiology

By
Dr. Befekadu Haile
(PhD/MSc in Plant Pathology; BSc in Biology)
 Chapter- one
1. Introduction

What is microbiology?

What are the branches of microbiology?

How have we gained our present knowledge of

microbiology?

Why is it such an important subject?

 1.1. Definition and Scope of Microbiology
 Microbiology :
 is a specialized area of biology that concerns with the study of
microbes ordinarily too small to be seen without magnification
 (Greek words; mikros - small + bios-life+ logos-to study).
 Size < 1 mm in diameter
 MOs. include diverse group of microscopic organisms
• exist as single cell or cell clusters (e.g., bacteria, archaea, fungi,
algae, protozoa and helminthes) and
• the viruses, which are microscopic but not cellular.
 Microbiology is concerned with the study of the
– structure
– Function
– Classification of microriganisms and
– with ways of controlling and using their activities
 Organisms included in the study of
Microbiology
1. Bacteria  Bacteriology

2. Protozoans  Protozoology

3. Algae  Phycology

4. Parasites  Parasitology

5. Fungi
– Yeasts and Molds  Mycology
6. Viruses  Virology
 Beneficial
 MOs are found
– required for the production of:
everywhere on earth, • bread
• cheese, yogurt,
which includes: • alcohol
– living creatures • antibiotics, vaccines
• vitamins, enzymes
(humans, animals,  Detrimental
plants) – Pathogenic (only a minority of
microorganisms)
– non-living creatures
(soil, water and  Microbiology helps us to understand
the fundamental nature and
atmosphere) characteristics of microbes
 Branches of Microbiology

1. Basic (pure) Microbiology:

• Fundamental nature of MOs is studied

• Microbial physiology- how the microbial cell functions biochemically

• Microbial taxonomy- The naming and classification of microorganisms

• Microbial ecology- The relationship between MOs and their ENV’T

• Microbial genetics- The study of genes of microbes

• Cellular microbiology: generally study about microbial cells

• Evolutionary microbiology: developmental process from earlier life

• Microbial cytology: cell structures and functions

 Branches of Microbiology…cont’d

2. Applied microbiology
– where the information learned from basic microbiology is employed for
control or use of the microorganisms
 It includes:
– Medical microbiology-
The study of the pathogenic microbes and the role of microbes in
human illness.
– Pharmaceutical microbiology
 The study of MOs that are related to the production of pharmaceutical
products and that cause pharmaceutical contamination and spoil.
– Industrial microbiology
• microbes for use in industrial processes
– Food microbiology
• MOs causing food spoilage and foodborne illness; and
• using microorganisms to produce foods
 Applied microbiology……cont’d

– Agricultural microbiology
 concerns with the study of agriculturally relevant MOs
studies relationships among microbes and crops/plants

 it deals with all aspects of farming and microbial

reactions in soil, diseases of plants

 emphasis on control of plant diseases and improvement

of yields
1.2. Historical Development of
Microbiology
 i. The First Observations

 MOs were discovered more than 300 years ago

 In 1665, Robert Hooke reported that living things were

 composed of little boxes or cells.

 Antony van Leeuwenhoek (1632-1723)

made first observation of MOs and
laid the foundation for bacteriology and protozoology.
 Antony Van Leeuwenhoek
 Father of microbiology who first properly described the different
shapes of bacteria

 Observed “animalcules” using simple microscope with one

lens.

11
 Edward Jenner (1749-1823)- English physician
 his great gift to mankind was his vaccine for smallpox

 a less pathogenic agent could confer protection against a more

pathogenic one (vaccines and vaccination)

 The modern era of vaccines and vaccination began in 1798 with

Edward Jenner’s
– use of cowpox as a vaccine against smallpox

 the nature of the virus was not known (b/c of infancy of

microscope)

 the mid 1800s- MB. became an experimental science

 MB.’s foundations were established in the later 19thc with the work
of Louis Pasteur and Robert Koch
 ii. The golden age of microbiology (1857–1914)
 Golden Era of Microbiology
 The period began in 1857 with the work of Louis Pasteur and
continued into the 20th C
– numerous branches of microbiology were laid for the maturing
process
– Dev’t of the Germ Theory of Disease
– Solution for the debate over Spontaneous Generation

 Louis Pasteur (1827-1895)- French chemist and microbiologist

 was the first to report the role of MOs in fermentation
(the role of yeasts in fermentation & role of bacteria for spoilage of wine)
 He firmly disproved the spontaneous generation doctrine
 He proposed germ theory of disease
 discovered the existence of anaerobic MOs
 Pasteurization
 Vaccination (anti rabies and anthrax vaccines)
 Josef Lister (1827-1912)-Surgeon and Medical Scientists

 developed a system of antiseptic surgery in 1867

the Father of antiseptic surgery
 In 1878, Lister studied the lactic acid fermentation of milk

 He developed a method for isolating a pure culture of Bacterium

lactis

 Metchnikoff (1845–1916)

coined the term “phagocytosis” which means “the eating of

cells”
notable contribution was on the Bacillus bulgaricus therapy
 Dr. Joshua Lederberg
noted for two landmark discoveries in bacterial
genetics:
bacterial conjugation and transduction
both laying foundations for:
genetic engineering,
modern biotechnology and
genetic approaches to medicine
 Carl Woese
is famous for defining the Archaea
(a new domain or kingdom of life) in 1977 by phylogenetic
analysis of 16S ribosomal RNA
1.2.1. Theories on the origin of life

Where did microorganisms originate??

 On the basis of the observation, two major theories were
formulated.

1. Theory of spontaneous generation

 This theory states that living things originated “spontaneously”
from non-living things.

 Aristotle: observed spontaneous existence of ;-

- fishes from dried ponds, when the pond was filled with rain.

- toads, snakes and mice in moist soil

-flies and maggots in manure and decaying flesh

 Theories…

2. Theory of biogenesis

 This theory states that life comes from pre-existing life.

 Francesco Redi: The first scientist who tried to set an experiment to

disprove the theory of spontaneous generation.
 Utilized jars containing meat. (Some were covered, some were not).

 Maggots appeared in uncovered jars  developed from flies

eggs.

Results not accepted for microscopic organisms.

 Francesco Redi…

covered meat

Exposed meat Maggots No maggots

 John Needham
 Introduced the first culture medium for microbial growth.
 Utilized infusion broth prepared by boiling meat, grain, etc. to
extract nutrients.
 Broth put in flasks, some were sealed with corks, and some were
not.
All flasks became cloudy. i.e. Turbidity indicated growth
He suggested that life originated spontaneously from nonliving
matters
 The spontaneous generation opponent didn’t accept his conclusion, they
said it could be due to entrance organisms from air or flasks, due to
improper seal.
 Lazzaro Spallanzani

 Repeated Needham’s experiments to disproof spontaneous

generation in microscopic life.

Boiled broth after placing in flasks.

Sealed flasks by plugging with solid stopper.

Results more consistent with Redi’s.

Occasionally sealed flask  cloudy

 Not accepted by spontaneous generation supporters, because they

said that heating may have destroyed, degraded “vital force” and
air was not allowed to enter.
 Louis Pasteur

 The scientist who disproved the theory of spontaneous

generation once and for all.
He designed a large curved flask i.e. Pasteur goose flask
(Swan-necked flask) and placed a sterile growth broth
medium.

Shape of flask allowed air in but trapped dust particles

which may contain microbes
Louis Pasteur…
 Louis Pasteur…

 Air freely moved through the tube, but dust particles were trapped
in the curved portion of the flask.
 And, no microbial growth was observed.
 Louis Pasteur…

 Therefore, Pasteur proved that microorganisms entered to the

broth with the air but they did not evolve spontaneously
 The Germ Theory of Disease
 Before the time of Pasteur:
effective treatments for many diseases were discovered by
trial and error
the causes of the diseases were unknown
 The germ theory of disease:
 The germ theory of disease states that a specific disease is
caused by a specific type of microorganism
 1835: Agostino Bassi showed a silkworm disease was caused by a
fungus
 1865: Pasteur believed that another silkworm disease was caused
by a protozoa
 Koch: discovered the cattle disease anthrax was caused by bacteria
now known as Bacillus anthracis
 The Germ Theory of Disease…

 Koch: established a sequence of experimental steps

directly relating a specific microbe to a specific disease

 The first proof that bacteria actually cause disease came

from Robert Koch in 1876.

 Koch’s postulates, used to prove that a specific microbe

causes a specific disease
 Robert Koch (1843-1910)- German Physician
 Koch’s postulates
1. the suspected pathogenic organisms should be present in all
cases of the disease & absent from healthy animals

2. the suspected organisms should be grown in pure culture.

3. cells from a pure culture of suspected organism should cause

disease in healthy animal

4. the organism should be re-isolated & shown to the same as

the original.

Koch’s postulates remain fundamental to pathology

Koch’s postulates
 Summary of Koch’s postulates
 A specific disease is caused by a specific organism

 The organism should be isolated and grown in the lab

into a pure culture

 When the artificially cultured organism inoculated into

healthy but susceptible animal it should produce
symptoms of the same diseases

 The organism should be re-isolated from artificially

infected animal and grown into pure culture in the lab
 Drawbacks (Limitations) of Koch’s postulate

 Not every microorganism associated with diseases

Example: Normal flora
 Many healthy people carry pathogens but do not exhibit symptoms
of the disease. i.e asymptomatic individuals were not considered.
 Opportunistic microorganisms can cause diseases only in immuno-
compromised individuals

 Not all diseases are caused by microorganisms

Example: Diabetes, Asthma, hypertension etc.

 Some microbes are very difficult or impossible to grow in the

laboratory in artificial media, such as most viruses and some bacteria
Example: Treponema pallidum  cause of Syphilis
Contributions of Koch’s postulates

Emphasizes the importance of lab cultivation of

microorganism in artificial growth media

Showed that a specific microbe has specific activity

Contributed to the development of pure culture

 Modern Developments in Microbiology
 Bacteriology is the study of bacteria.

 Mycology is the study of fungi.

 Virology is the study of viruses.

 Parasitology is the study of protozoa and parasitic worms.

 Immunology is the study of immunity.

 Vaccines and interferon's are being investigated to prevent and cure

viral diseases.

 Recombinant DNA technology or genetic engineering involves

microbial genetics and molecular biology.
 Vaccination

1796: Edward Jenner inoculated a person with cowpox

virus.
 The person was then protected from smallpox.

Called vaccination from vacca for cow

The protection is called immunity.

 The Birth of Modern Chemotherapy
Treatment with chemicals is chemotherapy.

Chemotherapeutic agents used to treat infectious disease can be

synthetic drugs or antibiotics.

Antibiotics are chemicals produced by bacteria and fungi that

inhibit or kill other microbes.
 Quinine from tree bark was long used to treat malaria.

1910: Paul Ehrlich developed a synthetic arsenic drug,

salvarsan, to treat syphilis.

1930s: Sulfonamides were synthesized.

 The Birth of Modern Chemotherapy…

1928: Alexander Fleming discovered

the first antibiotic.

He observed that Penicillium fungus

made an antibiotic, penicillin, that
killed Staphyloccus aureus.

1940s: Penicillin was tested clinically

and mass produced.
 Importance of microorganisms for mankind
1. Microbes and health:

 Microbes cause diseases both in plants and animals

 help to devise methods to prevent and identification of causative

microbe.
Streptomycin from Streptomycetes, Penicillin-Penicillium,
Bactracin - Bacillus etc

2. Agriculture and food production

– nutrient recycling: nitrogen and carbon - increases soil fertility

which boosts production and productivity

– food industry
 Importance of microorganisms …
3. Production of essential nutrients
 Microorganisms that live in the intestinal tract produce digestive
enzymes or other nutrients useful for nutrition
4. Production of O2 in the atmosphere
 50% of O2 on Earth from alegea cyanobacteria
5. Primary source of organic material
 Autotrophs - take up CO2 and convert it to organic matter; this is
called primary production
 cryptobiotic soil- turning sand into topsoil
 utilizing nitrogen, oxygen and carbon dioxide from the air and
using sunlight as a source of energy
 The Microbial
Importance World and Us …
of microorganisms
6. Replenishment of soil nitrogen
 Break down wastes and incorporate nitrogen gas from the air into
organic compounds.

 By recycling chemical elements between the soil, water, life, and air

7. Biodegradation:
 Bacteria degrade organic matter in sewage.
 Bacteria degrade or detoxify pollutants
such as oil and mercury
 The Microbial
Importance World and Us …
of microorganisms

8. Source of antibiotics and drugs

9. Manufacture of fermented foods and beverages

10. Biological Insecticides

 Microbes that are pathogenic to insects are alternatives to chemical
pesticides to prevent insect damage to agricultural crops and
disease transmission.
• Bacillus thuringiensis infections are fatal in many insects but
harmless to other animals including humans and to plants.
Development of pure culture

What is pure culture???

 Pure Culture Methods
 Pure culture-

the one that contains only a single species of microbe

 A pure culture is sometimes called an axenic culture

 Obtaining pure culture is most important laboratory procedures

in microbiology

 Sterilization of growth medium

 The most commonly used methods of pure cultures in routine
laboratory work:
Pour plate method,
Streak plate method, and
Spread plate method
 Culture Media
Culture- any growth or cultivation of microorganisms.
Culture media is a growth media which support the growth of
MOs or cells in the laboratory.
 For successful isolation:
• Choice of the culture medium is very important
Some media (selective media) inhibit the growth of unwanted
organisms.
• Nutrients-
Nutrients must be added to the medium to support growth.
These usually include a carbon source (carbohydrate) such as
glucose, a nitrogen source like amino acids or nitrogen salts,
vitamins such as thiamine or biotin and minerals.
• The pH of the medium is also important
 For example, a pH below 4.5 inhibits the growth of most bacteria.
 Types of Culture Media
Culture media can be classified based on:
physical states
chemical composition
functional properties
 Culture media classified based on physical States
 Liquid Media (Broth): These media do not contain any
solidifying agent.
 Solid Media: Contain solidifying agent such as agar, silica gel or
gelatine
E.g. Nutrient agar
 .
Culture media classified based on physical States….

Semisolid Media:
• Fall in between liquid and solid media

• they are more jellylike than solid media

• Contain lower % of solidifiers

• useful in determining whether certain bacteria are motile or not

 Solidifying agent
Agar:
 derived from certain marine algae

 gels the medium

 permits easy observation

 keeps fungal or bacterial colonies separate

 permits diffusion of O2 and dissolved nutrients

 A culture medium solidified by agar is used for isolation of MOs
 Most media contain 15-20 gms agar/ liter
 Media containing agar will melt at about 95 ○C and gel at about 40 ○C
 Solidifying agent…
Gelatin
Gelatin is a protein source and solidifying agent for use in
preparing microbiological culture media.

sets at about 25°C but is liquefied by proteolytic bacteria

Silica gel
Silica is a desiccant (drying agent) - it absorbs moisture
from its surroundings.

form of silicon dioxide (silica)

is tedious to prepare.