INTRODUCTION TO

MICROBIOLOGY
MARY LOU DULLONA-BASA
 MICROBIOLOGY
 Study of living things too small to be seen without magnification
 Living things – microorganism or microbes - less than 1mm in diameter
 Bacteria
 Algae
 Protozoans
 Viruses
 Fungi

 Can only be viewed by a microscope

 Some organisms CAN be visualized without the aid of amplification
[bread molds (fungus) and filamentous algae
 Techniques necessary to isolate and culture microorganisms:
 Isolation
 Sterilization
 Culture in artificial media
Branches

 General microbiology: broad range of microbiological questions

 Medical microbiology: microbes that cause human disease
 Public health and epidemiology: Studies and controls transmission,
frequency, and distribution of disease
 Immunology: the immune system
 Agricultural microbiology: impact of microbes on agriculture
 Microbial ecology: relationships between microbes and their habitats
 Food microbiology: Prevention of food borne disease; microbes that make food
and drink
 Industrial microbiology: commercial use of microbes to produce products
 Biotechnology: manipulation of organisms to form useful products
 Biotechnology: harness of cellular and biomolecular processes to develop
technologies and products to improve life and health
 Epidemiology study of how often and why diseases occur in
different groups of people
 Bioremediation use of microorganisms, plants or microbial r plant
enzymes to detoxify contaminants in soil and other
environments
 Microbiology may be interested in specific types of organisms:
 Virology - viruses
 Bacteriology - bacteria
 Phycology - algae
 Mycology - fungi
 Protozoology – protozoa
 Parasitology – parasitic protozoans and helminths
 Microbiologists may have a more applied focus:
 Medical microbiology, including immunology
 Food and Dairy microbiology
 Public Health microbiology (Epidemiology)
 Industrial microbiology
 Agricultural microbiology
 Microbiologists may be interested in various characteristics or
activities of microorganisms:
 Microbial morphology
 Microbial cytology
 Microbial physiology
 Microbial ecology
 Microbial genetics and molecular biology
 Microbial taxonomy
Importance of Microbiology
 Keeping the planet healthy
 Microbes are essentially protectors of the planet, ensuring that minerals such as carbon and
nitrogen are incessantly recycled. They play a crucial role in keeping the atmosphere oxygenated
and also actively degrade dead organic matter, thus transforming organic carbon back into carbon
dioxide.
 Agriculture
 When it comes to creating soils that can support crops and livestock, microbes are indispensable
Studying microbiology helps farmers to optimise nitrate levels and maximise output.
 Combating disease
 Infectious diseases have the capacity to wipe out entire populations, and microbiology is the key
to keeping outbreaks under control. The study of microscopic organisms allows scientist to
develop antibiotics and vaccines, with revelations such as Alexander Flemming’s discovery of
penicillin saving millions upon millions of lives.
 Chemical products
 From antibiotics and solvents to preservatives and pharmaceuticals, microbes are used to create a
myriad of useful products that we take for granted. Uncovering these chemical reactions and
retailing them as commercial goods shapes the face of life as we know it.
 Biotechnology
 Genetic engineering is an incredibly exciting revelation, and microbiology lies at the heart of the
sphere. The scientific process of freely moving genes from one organism to another, isolating
DNA and manipulating results is all hosted by bacteria. 
 Microbiology has come a long way since it was first pioneered by Dutch draper and
hobbyist glass grinder Antonie Van Leeuwenhoek in the 17th century. 
‘A ‘Dyeing’ Art in Microbiology’ explores how the “Father of Microbiology" led us to
discover the staining methods currently used in cytology, histology, haematology,
microbiology and parasitology, 
Microorganisms - microbes

 There are two types of microorganisms:

 Prokaryotes
 have a relatively simple morphology and lack a true
membrane-bound nucleus
 Eukaryotes
 are morphologically complex and have a true, membrane-
bound nucleus
The Microbes
Size of Microbes
Beneficial Uses of Microbes

Synthesis of drugs, hormones and enzymes

Beneficial Uses of Microbes

Bioremediation is the use of microbes to degrade organic

matter in sewage and detoxify pollutants such as oil spills.
Modern Uses of Microbes

 Biotechnology, the use of microbes as miniature biochemical

factories to produce food and chemicals is centuries old.
 Genetic engineering makes use of molecular biology and
recombinant DNA techniques as new tools for biotechnology.
 Gene therapy replaces missing or defective genes in human cells
through genetic engineering.
 Genetically modified bacteria are used to protect crops from pests
and freezing.
 Five Kingdoms of Life -Robert H. Whittaker

 Organisms can be divided into five kingdoms:

 the Monera or Procaryotae,
 Protista,
 Fungi,
 Animalia, and
 Plantae
Microbial Taxonomy – Woese-Fox

 Organisms are
divided into
three domains
 Bacteria
 Archae
 Eukarya
HISTORICAL PERSPECTIVE

 ROBERT HOOKE
 Crude microscope – 1665
 Most important discovery of biology

 Cell – smallest structural unit of life

 ANTONY VAN LEEUWENHOEK
 First to observe living microbes
 His single-lens magnified 50-300X magnification
 Between 1674-1723 he wrote series of papers describing his observations of bacteria, algae,
protozoa, and fungi (Animalcules)
SPONTANEOUS GENERATION

 abiogenesis
 some forms of life could arise from “vital
forces” present in nonliving or decomposing
matter
 organisms can arise from non-living matter.
 LOUIS JABLOT
 1670 - conducted an experiment in which he divided a hay
infusion that had been boiled into two containers:
 a heated container that was closed to the air
 a heated container that was freely open to the air
 Only the open vessel developed microorganisms
 helped to disprove abiogenesis.
REDI’S and JABLOT’S EXPERIMENTS
 Disproved by:
 Schwann, Friedrich Schroder and von Dusch (1830s)
 Air allowed to enter flask but only after passing through
a heated tube or sterile wool
 John Tyndall (1820-1893) – Omission of dust  no growth.
 Demonstrated heat resistant forms of bacteria
(endospores)
LOUIS PASTEUR (1822 - 1895)

 Disproved spontaneous generation of microbes by preventing

“dust particles” from reaching the sterile broth
 In 1861 completes experiments that lays to rest spontaneous
generation.
 Showed microbes caused fermentation
and spoilage
 PASTEUR’S EXPERIMENT

trapped airborne organisms in cotton; he also heated the necks of

flasks, drawing them out into long curves, sterilized the media, and
left the flasks open to the air.
In this way Pasteur disproved the theory of spontaneous generation
Role of Microorganisms in Disease

 Demonstrations that micoorganisms cause disease

 Oliver Holmes (1773 - 1843)
 showed that sepsis could be transmitted by hands of medical student and
may cause disease
 M. J. Berkeley (ca. 1845)
 demonstrated that the Great Potato Blight of Ireland was caused by a
Fungus
 Louis Pasteur (1822 - 1895)
 showed that the pébrine disease of silkworms was caused by a protozoan
parasite
 Edward Jenner (ca. 1798)
 Develop the first Vaccine and used a vaccination
procedure to protect individuals from smallpox
 Louis Pasteur
 developed other vaccines including those for chicken
cholera, anthrax, and rabies
 1796 – First vaccine (smallpox) by Edward Jenner
 1885 - Vaccine against Rabies Louis Pasteur
 Robert Koch (1843 - 1910),
 using criteria developed by his teacher, Jacob Henle
(1809-1895), established the relationship between
Bacillus anthracis and anthrax.
 His criteria became known as Koch’s Postulates and
are still used to establish the link between a
particular microorganism and a particular disease:
 1884 Koch’s Postulates of Disease Transmission -
Robert Koch
Koch’s Postulates

 The causative (etiological) agent must be present in all

affected organisms but absent in healthy individuals
 The agent must be capable of being isolated and
cultured in pure form
 When the cultured agent is introduced to a healthy
organism, the same disease must occur
 The same causative agent must be isolated again from
the affected host
Development of Culture Media

 To enable the isolation of pure cultures (only one type of organism)

 Especially important during Koch’s period
 Gelatin not useful as solidifying agent (melts at >28 ºC and some bacteria
hydrolyze it with enzymes)
 Fannie Hesse, the wife of one of Koch’s assistants, proposed using agar
 Not digested by most bacteria
 Melts at 100 ºC
 Used today - ~2% in solid media
 Richard Petri, another of Koch’s assistants, developed the Petri dish
ROBERT KOCH

 Developed pure culture methods.

 Identified cause of anthrax ( Bacillus anthrax) , TB (Mycobacterium
tubercullosis) , & cholera ( Vibrio cholera).
Development of Vaccines and Antisera

 Vaccination:
 Inoculation of healthy individuals with weakened (or attenuated) forms of microorganisms,
that would otherwise cause disease, to provide protection, or active immunity from disease
upon later exposure.

 Edward Jenner in 1796 discovered that cowpox (vaccinia) induced protection against
human smallpox
 Called procedure vaccination
 Pasteur and Roux reported that incubating cultures longer than normal in the lab
resulted in ATTENUATED bacteria that could no longer cause disease.
 Working with chicken cholera (caused by Pasteurella multocida), they noticed that animals
injected with attenuated cultures were resistant to the disease.
 Pasteur and Chamberland developed other vaccines:
 Attenuated anthrax vaccine
 Chemical and heat treatment (potassium bichromate)

 Attenuated rabies vaccine

 Propagated the virus in rabbit following injection of infected brain and spinal cord extracts
 Passive immunization
 Work by Emil von Behring (1845-1917) and Shibasaburo Kitasato
(1852-1931)
 Antibodies raised to inactivated diphtheria toxin by injection different host (rabbit) with the toxin
(a toxoid form)

 Antiserum recovered
 Contains antibodies specific for the toxin
 Protection from disease when injected non -immune subject.
 JOHN TYNDALL (1820 – 1893)
 In 1876 discovered that there were two different types of bacteria.
 a) Heat sensitive or heat labile forms (vegetative cells)
easily destroyed by boiling
 b) Heat resistant types known as an endospore
 Tyndall demonstrated that alternate process of heating & cooling
if repeated five times, can kill all the endospores.
 This is known as Sterilization process or Tyndallization
FERDINAND COHN

 In 1876, a German botanist, Ferdinand Cohn, also

discovered “heat-resistant forms of bacteria”. This
bacteria are now termed endospores.( Bacillus species and
Clostridium species)
 Anthrax bacillus
GOLDEN AGE OF MICROBIOLOGY

 1860 to 1900
 Golden Age of Microbiology
 rapid advances, spear-headed by Louis Pasteur
and Robert Koch
 led to the establishment of microbiology as a
science.
LOUIS PASTEUR - 1822 – 1895

 1864
 Pasteurization
 established the relationship between microbes and disease in preventing wine
from spoiling
 kills bacteria in the alcohol by heat, thus preventing the formation of acetic acid
(vinegar).
 His discovery of pasteurization, lead Pasteur to introduce the “germ theory of disease”
in 1864. Pasteur stated that diseases are caused by the growth of microbes in the body
and not by sins, bad character, or poverty, etc.
 Joseph Lister (1827 - 1912)
 1867 Antiseptic Surgery( Carbolic acid- Phenol)
 developed a system of surgery designed to prevent
microorganisms from entering wounds – phenol (Carbolic
Acid) sprayed in air around surgical incision
 Decreased number of post-operative infections in patients
 his published findings (1867) transformed the practice of surgery
PAUL EHRLICH

 1890’s - Ehrlich proposed a theory of immunity in which

antibodies were responsible for immunity( Antitoxin)
 father of modern chemotherapy
 speculated about some “magic bullet” that would selectively find
and destroy pathogens but not harm the host (Selective Toxicity)
 develop a staining procedure to identify tubercle bacilli.
ALEXANDER FLEMING

 1928 - Fleming observed that the growth of the bacterium

staphyloccus aureus was inhibited in the areas surrounding the
colony of a mold that had contaminated a Petri plate
 mold was identified as Penicillium notatum, and its active
compound was named penicillin.
 1929
 discovery of Penicillin - first antibiotic
 Walter Hesse ( 1846-1911): Used Agar as a solidifying agent to
harden media. Agar is extracted from seaweeds red algae.
 Rechard Petri ( 1852-1921): Used agar dish to provide a large
area to grow.
 Christian Gram ( 1853-1935): Staining method that demonstrate
bacteria and distinguish between Gram positive and Gram
negative bacteria.
 Raymond Sabouraud ( 1890-1910): Develop culture media to
study yeast and molds.
 Dimitri Ivanovski (1892): Tobacco mosaic virus could pass
through filters used to remove bacteria.
 Selman Waksman (1940): Discovered a number of antibiotic such
as Tetracycline and Streptomycin.
WATSON and CRICK, FRANKLIN, and
WILKINS
 1953 - Watson and Crick determined the structure of DNA
 used their research, together with the research of Franklin and
Wilkins to determine the structure of the DNA molecule.
Microorganisms in the Early 20Th Century
Important Early Discoveries
 George W. Beadle and Edward L. Tatum (ca. 1941)
 studied the relationship between genes and enzymes using the
bread mold, Neurospora
 Precursor ornithine  citrulline  arginine
 One gene, one polypeptide hypothesis
 Salvadore Luria and Max Delbruck (ca. 1943)
 Demonstrated spontaneous gene mutations in bacteria (not
directed by the environment)
 Oswald T. Avery, Colin M. MacLeod, and Maclyn McCarty (1944)
 Following initial studies by Frederick Griffith (1928) they
provided evidence that deoxyribonucleic acid (DNA) was the
genetic material and carried genetic information during
transformation
 Worked with Streptococcus pneumoniae (rough and smooth)
 In the 1970s new discoveries in microbiology led to the development
of recombinant DNA technology and genetic engineering
 THE MICROSCOPE
 The development of magnifying instruments enabled scientists to learn more about microorganisms
 Compound light microscope
 Use series of lenses
 Use glass slides
 Magnification – 1000-2000X
 Uses light
 Transmission electron microscope
 Much better resolution- better details
 100000 X magnification
 Uses rays of electrons
 Uses photographic plate for viewing
 Setback-cant use living cells
 Scanning electron microscope
 Coolest
 Enables to view in 3D image
 uses a beam of electrons moving at low energy to focus and scan specimens
 Reska (1938) – First Electron
Microscope
 The electron microscope is capable
of magnifying biological
specimens up to one million times.
These computer enhanced images
of smallpox, herpes simplex, and
mumps are magnified,
respectively, 150,000, 150,000 and
90,000 times.
 To study detail structures of viruses.
Domains

 The highest – largest category, recent addition

 3 domains
 1. Archaea – ancient “bacteria”, unicellular like bacteria, also simple
cell structure (prokaryote – no nucleus) but have distinct metabolism
(chemistry) allowing them to exist in “extreme” environments
 2. Bacteria – unicellular, prokaryote, found everywhere (Old
kingdom name – Monera)
 3. Eukarya – unicellular to multicellular, complex and organized
cells with nuclei and organelles (mitochondria)
 Domain Bacteria
Prokaryotes

Peptidoglycan cell walls

Binary fission
QuickTime™ and a
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For energy, use of organic
chemicals, inorganic
chemicals or
photosynthesis
Ex: Escherichia coli
 Domain Archaea

Prokaryotes
Lack peptidoglycan
Live in extreme environments
(extremophiles)
Include:
 Methanogens
 Extreme halophiles
 Extreme thermophiles
Domain Eukarya

 The cells of all eukaryotes have a membrane-bound nucleus. Members of the

Domain Eukarya are further categorized into one of four Kingdoms.
 Fungi

Eukaryotes

Chitin cell walls

Molds and mushrooms QuickTime™ and a

TIFF (Uncompressed) decompressor
are multicellular are needed to see this picture.

 Mycelia – hyphae -
filaments

Yeasts are unicellular

 Protozoa

Eukaryotes

Mostly saprobes and

commensals QuickTime™ and a
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May be motile by means
of pseudopod, cilia or
flagella

Absorb or ingest organic

chemicals
 Algae

Eukaryotes
Cellulose cell walls
Photosynthetic QuickTime™ and a
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 Primary producers
Produce molecular oxygen
and organic compounds
Part of food chain
 Helminths

Eukaryotes
Multicellular animals
Parasitic flatworms and QuickTime™ and a
roundworms called TIFF (Uncompressed) decompressor
are needed to see this picture.
helminths
Microscopic stages in life
cycles
 Viruses

Acellular
Obligate intracellular parasites
Genome consist of DNA or
RNA called Core QuickTime™ and a
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Core surrounded by protein
coat called Capsid
Virion (coat) may be enclosed
in lipid envelope
Replicated only when they are
in a living host cell
 Prions

Proteinaceous infectious
agents
Causes Bovine Spongiform
Encephalopathy (BSE) QuickTime™ and a
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are needed to see this picture.

Also causes Creutzfeldt-

Jacob Disease (CJD)
An Emerging Infectious
Disease (EID)
 Infection and Disease
 Infection the entry of a microbe into the host.

 Disease infection followed by the appearance of signs and symptoms.

 Pathogen an infectious or disease agent.

 Saprobe a microbe that lives on dead or decaying organic matter.

 Opportunistic pathogen
is a microbe that cause disease in immunocompromised hosts or when the
normal microbiota is altered.
Emerging Infectious Diseases

 Occurrence of new diseases and increasing incidence of old ones (EID)

 Factors:
a) evolutionary changes in existing organisms
(b) spread of known diseases into new geographic areas by
modern transportation
(c ) ecological changes resulting in introduction of unusual agents
(d) emergence of antimicrobial resistance
Emerging Infectious Diseases

 West Nile Encephalitis, first diagnosed in Uganda in 1937; appeared in New York
City in 1999.
 Invasive Group A Streptococcus, also known as the “flesh eating bacteria”
 Escherichia coli 0157:H7, causes “bloody diarrhea” and hemorrhagic uremic
syndrome (HUS)
 Bovine Spongiform Encephalopathy (BSE) or “mad cow” disease caused by prions
 Acquired Immunodeficiency Syndrome (AIDS) caused by HIV and Africa is
hardest hit
 Anthrax caused by Bacillus anthracis was sensationalized in 2001 when spores were
disseminated via the mail