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From the Virtual Microbiology Classroom on ScienceProfOnline.com Image: Compound microscope objectives, T. Port
 History of
Microbiology

Image:
From the Virtual Microbiology Classroom on ScienceProfOnline.com Drawing of Death Bringing Cholera in
 Spontaneous Generation Debate

 The belief in the spontaneous

generation of life from
nonliving matter was introduced
by Aristotle, who lived around
350 BC.

 According to Aristotle, it was:

“readily observable that aphids arise from the
dew which falls on plants, fleas from putrid
matter, mice from dirty hay.”

 This belief remained

unchallenged for more than
2000 years.

 Until… Aristotle: 384 – 322 B.C.

Images: Aphids, Flagstaff Fotos;

Bust of Aristotle by Lysippus, Wiki From the Virtual Microbiology Classroom on ScienceProfOnline.com
 Spontaneous Generation Debate

Francesco Redi - Experiments on Flies

First to formally challenge the accepted belief of spontaneous generation.

Redi's Question: Where do maggots come from?

Hypothesis: Maggots come from flies.

Experiment: Redi put meat into three separate jars.

Jar-1
• Left open
• Maggots developed
• Flies were observed laying eggs on the meat in the open jar

Jar-2
• Covered with netting
• Maggots appeared on the netting
• Flies were observed laying eggs on the netting Francesco Redi, Italian physician,
naturalist & poet, 1626 – 1697.

Jar-3
• Sealed
• No maggots developed Images: Redi’s experiment , Unknown;
From the Virtual Microbiology Classroom on ScienceProfOnline.com Fracesco Redi portrait. Wiki
 Spontaneous Generation Debate 1632 - 1723

Anton van Leeuwenhoek’s “Animalcules”

(Pronounced Lay-ven-hook)

 As a draper (merchant who sells cloth and dry goods) , he

used lenses to examine cloth. This probably
led to his interest in lens making.

 He assembled hundreds of microscopes, some

of which magnified objects 270 times.

 As he looked at things with his microscopes, he

discovered “micro” organisms - organisms so
tiny that they were invisible to the naked eye.

 He called these tiny living organisms

“animalcules”. He first described bacteria,
protozoans and many cells of the human body.

LISTEN to the Radiolab episode “Sperm” to

learn how Anton let his freak flag fly!

♬ Sing It SUPER FREAK!! ♬

Images: Leeuwenhoek portrait 1680, Jan Verkolje (I);Replica

of Leeuwenhoek microscope, Jacopo Werther; Man with scope, source
unknown; Background image is sperm that van Leeuwenhoek drew. From the Virtual Microbiology Classroom on ScienceProfOnline.com
 Anton van Leeuwenhoek: “Animalcules”
(Pronounced Lay-when-hook)

Watch Video:
Below is a poem about Van Leeuwenhoek by Maxine Kumin, from the “Pond Life Under the Microsco
fantastic book of science-related poetry The Tree That Time Built. pe”
The Microscope
Anton Leeuwenhoek was Dutch. He sold pincushions, cloth, and such.
The waiting townsfolk fumed and fussed, as Anton’s dry goods gathered
dust.

He worked, instead of tending store,

At grinding special lenses for
A microscope. Some of the things
He looked at were: mosquitoes’ wings,
the hairs of sheep, the legs of lice,
the skin of people, dogs, and mice;
ox eyes, spiders’ spinning gear,
fishes’ scales, a little smear
of his own blood, and best of all,
the unknown, busy, very small
bugs that swim and bump and hop
inside a simple water drop.

Impossible! Most Dutchmen said.

This Anton’s crazy in the head!
We ought to ship him off to Spain!
He says he’s seen a housefly’s brain!
He says the water that we drink
Is full of bugs! He’s mad, we think!

They called him dumkopf, which means dope.

That’s how we got the microscope.

From the Virtual Cell Biology Classroom on ScienceProfOnline.com
 The Controversy Over Spontaneous Generation
John Needham & Lazzaro Spallanzani

The Question:
French
What causes tiny living things to appear in decaying broth? chemist
Louis Pasteur’s
design of this
Needham’s Hypothesis: Spontaneous generation. experiment settled
the argument.
Spallazani’s Hypothesis: Microbes come from the air. Boiling will kill them.

Needham >

1713 - 1781

Spallazani >

1729 - 1799

Image: Louis Pasteur, Pierre Lamy Petit circa

1866; Lazzaro Spallanzani, 1729-1799 From the Virtual Microbiology Classroom on ScienceProfOnline.com
 The Controversy Over Spontaneous Generation
Louis Pasteur & Industrial Microbiology Pasteur’s
Observations:
1. Blobs were alive because
- Q: What is fermentation? they divide and make more of
themselves.
- What causes fermentation?
Some scientists thought that air caused fermentation 2. Put grape juice + yeast in open and
in air-tight containers. Fermentation
Others thought that microbes caused fermentation. occurred in both. This means that
yeast are facultative anaerobes.
- Q: What is pasteurization?
3. Took two flasks of sterile grape juice
and introduced bacteria into one and
yeast into another.
Are these non-living
blobs or living
microbes?
< yeast + grapes = yummy wine  (ethanol)
bacteria + grapes = spoiled wine  (lactic acid) >

Image: Louis Pasteur, Pierre Lamy Petit, circa 1866; Grapes fermenting;
Sachromyces yeast; Lactobacillus, Public health Image Library #1048 From the Virtual Microbiology Classroom on ScienceProfOnline.com
 REVIEW!
Animated lesson of experiments testing
Spontaneous Generation

Images: Redi’s experiment ,

From the Virtual Microbiology Classroom on ScienceProfOnline.com Unknown; Swan neck flask, Wiki
 The Germ Theory of Disease
Sherlock This…

 Oliver Wendell Holmes Sr. & Ignaz

Semmelweis - physicians practicing in
different parts of the world.

 Holmes (US)
Believed death following childbirth (puerperal fever)
often caused by the material on hands of midwives or 1809 - 1894
attending physicians.

 Semmelweis (Austria)
Noticed death rates higher in maternity wards staffed
by medical students than in those attended by midwives.
Death rates decreased in summer.

1818 - 1865

Q: Why?
Images on right: Ignaz Semmelweiss, 1860
From the Virtual Microbiology Classroom on ScienceProfOnline.com Jeno Doby; Oliver Holmes Sr., 1883
 The Germ Theory of Disease
Importance of Hand Washing

Sung to tune of "Row, row, row your boat."

Wash, wash, wash your hands,

Play our handy game.

Rub and scrub, scrub and rub,

Germs go down the drain.

(Sing 2x = about 20 seconds.)

Images: Hand washing diagram, used

From the Virtual Microbiology Classroom on ScienceProfOnline.com with permission © Clinical Skills Ltd
 Meet the Microbe!
• Organism: Streptococcus
pyogenes
Streptococcus

Streptococcal Infections

Streptococcus is a Gram + cocci-shaped genus of

bacteria, which produce toxins that contributes to its
pathogenesis.

Some diseases caused by this bacterium include:

• Puerperal fever
• Strep Throat
• Streptococcal Pneumonia
• Scarlet fever
• Necrotizing fasciitis (flesh-eating bacteria)

Images: Scarlet fever strawberry tongue Public Health Image

Library (PHIL) # 5120; Streptococcus pyogenes, PHIL #2110;
Necrotizing fasciitis, Smuszkiewicz, Trojanowska & Tomczak. From the Virtual Microbiology Classroom on ScienceProfOnline.com
 Germ Theory of Disease
1813 - 1858

Dr. John Snow & the Investigation of Cholera

 Played key role in setting standards for good
public hygiene and preventing spread of
infectious disease.

 Snow skeptic of the then-dominant “miasma

theory” (disease caused by bad air).

 Believed cholera transmitted by water

contaminated with waste of other cholera
sufferers.

 Mapped occurrence of cholera cases during

epidemic in London and found cases centered
around a specific public water supply.

 Asked city of London to dismantle the pump.

 Q: What do you think happened once water

pump was dismantled?

Images: Snow’s map of London, published by C.F.

Cheffins, 1854;. John Snow, 1813-1858, Wiki From the Virtual Microbiology Classroom on ScienceProfOnline.com
 Disease, Please! Cholera
• Infectious gastroenteritis caused by the Gram
- bacterium Vibrio cholerae.

• Transmission occurs through ingesting

contaminated water or food.

• Action on mucosal epithelium lining of the small

intestine responsible for the characteristic
massive diarrhea.

• One of the most rapidly fatal illnesses known.

• Can progress from first liquid

stool to shock in 4 to 12 hours,
with death quickly following
without rehydration treatment.

• Read excerpt from

• “The Dress Lodger” p.114

Image: Drawing of Death Bringing Cholera in Le Petit

Journal, circa 1912; Vibrio cholerae, PHIL #5324 From the Virtual Microbiology Classroom on ScienceProfOnline.com
 Germ Theory of Disease
John Tyndall and the Discovery of Endospores
 Discovered that some bacteria existed in two forms:
1. heat-stable form (endospore)
2. heat-sensitive form (vegetative cell)

 Need prolonged or intermittent heating to destroy

the heat-stable endospores.
1820 - 1893

 His research resulted in a method of sterilizing

liquid by heating it to boiling point on successive
days, referred to as Tyndallization.

 Tyndallization is useful for sterilization of growth

media in science classes and other situations where
autoclaves not available for pressure sterilization.

Variations in
endospore morphology:
(1, 4) central
endospore; (2, 3, 5)  
terminal endospore;
(6) lateral endospore

Images: John Tyndall; , H.B. Hall N.Y.

From the Virtual Microbiology Classroom on ScienceProfOnline.com 1878; Endospore stain T. Port
 Bacterial Genus : Clostridium
GRAM-POSITIVE
Obligate anaerobes
bacillus-shaped
endospore producer

 The members of this genus have a couple of bacterial

“superpowers” that make them particularly tough
pathogens.

 All have a strictly fermentative mode of metabolism

(Don’t’ use oxygen).

 Vegetative cells are obligate anaerobes killed by

exposure to O2, but their endospores are able to
survive long periods of exposure to air.

 Known to produce a variety of toxins, some of which

are fatal.
- Clostridium tetani = agent of tetanus

- C. botulinum = agent of botulism

- C. perfringens = one of the agents of gas gangrene

- C. difficile = part of natural intestinal flora, but resistant

strains can proliferate and cause pseudomembranous colitis.

Images: Man with Tetanus, Sir Charles Bell; Clostridium

botulinum, PHIL #2107; Wet Gangrene, Wiki From the Virtual Microbiology Classroom on ScienceProfOnline.com
 Germ Theory of Disease
Robert Koch
1843 - 1910

 Experimented with medium to grow

bacteria on.

 He tried gelatin, but it did not work.

 Wife of colleague recommended agar (a

gelatin-like product derived from
seaweed).

 Didn’t melt, and bacteria couldn’t digest it.

 He could also add various nutrients

necessary to grow certain organisms.

 Koch (pronounced Coke) originated use of a two

part dish for growing bacteria ( Petri dish
named after Julius Petri, a German
bacteriologist), and a technique for
isolating pure bacterial colonies.

Images. Micrococcus luteus colonies, T. Port; Robert Koch portrait,

From the Virtual Microbiology Classroom on ScienceProfOnline.com 1843-1910, NIH; MAC differential media, T. Port
 Disease, Please!
Anthrax
Gram + bacteria
Bacillus anthracis

 An endospore-producing bacterium.
(Genera Bacillus & Clostridium examples of
Robert Koch's original micrographs of the anthrax bacillus.
endospore producing bacteria.)

 Bacillus anthracis first bacterium

proven to be the cause of a disease.

 Anthrax was a disease killing European livestock. Farm animals,

apparently healthy in the morning, might die by the end of the day,
blood turned black.

 Humans interacting with the animals were also at risk of becoming

ill.

 In 1877, Robert Koch grew Bacillus anthracis in pure culture,

demonstrated its ability to form endospores, and produced
experimental anthrax by injecting it into animals.

 These experiments resulted in Koch formulating guidelines, called

Koch’s Postulates, for linking specific organisms with specific
diseases.

Image: Gram stained Bacillus anthracis, ,Public Health Image

From the Virtual Microbiology Classroom on ScienceProfOnline.com Library #2105; Inhilation Anthrax in monkey spleen tissue, NIH.
Germ Theory of Disease
Koch’s Postulates

REVIEW!
Click through
animated lesson o
n Koch’s Postulate
s
connecting
Bacillus anthracis
with the disease
anthrax.

From the Virtual Microbiology Classroom on ScienceProfOnline.com

 Germ Theory of Disease
Gram Stain
 First of Koch’s postulates demands that the suspected agent must be found in every case of a
given disease.
 That means the tiny microbes must be seen and identified. However, in most cases, microbes are
colorless and difficult to see.

 Christian Gram (1850-1938) developed a technique, the Gram stain, that is still widely used today.

 Differential stain that involves the application of a series of dyes.

 Leaves some microbes purple and others pink.

 Microbes that stain purple, Gram-positive, and those that stain pink, Gram-negative.

From the Virtual Microbiology Classroom on ScienceProfOnline.com Images: Gram positive and Gram negative bacterial stains, T. Port
 The Germ Theory of Disease
Dr. Joseph Lister – Father of Modern Antisepsis
Prevailing belief = Wound infection due to exposed to stinking "miasma" in air.

Lister had read a paper by Pasteur showing that rotting could occur without
oxygen if microorganisms present.
1827 - 1912
Maybe microorganisms were causing gangrene?

Pasteur suggested three methods to get rid of microorganisms:

1. ________________________________

2. ________________________________

3. ________________________________

The first two inappropriate for use in human wounds, so Lister experimented with the third.
Carbolic acid was used for deodorizing sewage, so Lister tried spraying instruments, surgical
incisions, wounds and dressings with solution.

It markedly reduced the incidence of gangrene.

As germ theory became more widely accepted, understood infection best avoided by preventing
bacteria from getting into wounds in the first place. This led to the rise of sterile surgery.

From the Virtual Microbiology Classroom on ScienceProfOnline.com Image: Joseph Lister, circa 1860, Wiki
 The Germ Theory of Disease
Florence Nightingale and Nursing

 English nurse who was a

proponent of cleanliness and
antiseptic techniques to the 1820 - 1910
field of nursing.

 She came to prominence for her

pioneering work in nursing during
the Crimean War (say cry-me-in),
where she tended to wounded
soldiers.

 She was meticulous about

collecting and analyzing data on
her practices, to provide proof
of their efficacy.

 Founded the Nightingale School

for Nurses, the first nursing
school in the world.

Image: Florence Nightingale from Carte de Visite, circa

From the Virtual Microbiology Classroom on ScienceProfOnline.com 1850; Polar area diagram by Florence Nightingale 1858.
 Discovery of Antimicrobial Agents
Sulfa Drugs (Sulfonamides)

• Dr. Gerhard Domagk (say Doe-mock), a

German chemist, discovered that the 1895 - 1964
dye Prontosil was effective against a
wide range of bacteria.

• Sulfanilamide portion of the Prontosil

molecule is responsible for its
antibacterial effect. Inhibits essential
enzyme activity in effected bacteria.

• Won the 1939, Nobel Prize in Medicine.

Images: Dr. Gerhard Domagk, 1964,

From the Virtual Microbiology Classroom on ScienceProfOnline.com Wiki, Sulfanilamide elixer, Source
 Discovery of Antimicrobial Agents
Penicillin

 Alexander Fleming (1881 – 1955),

a Scottish biologist and
pharmacologist, observed bacterial
staphylococci colonies disappearing
on plates contaminated with mold.

 Fleming extracted the compound

from the mold responsible for
destruction of the bacterial
colonies.

 The product of the mold was

named penicillin, after the
Penicillium mold from which it was
derived.

 Nobel Prize in Physiology of

Medicine in 1945.
Images: Penicillium mold, PHIL #8396; Staphylococcus aureus
on antibiotic test plate, PHIL #2641;
Poster attached to a mailbox offering advice to World War II
From the Virtual Microbiology Classroom on ScienceProfOnline.com servicemen
 Discovery of Antimicrobial Agents
Antibiotic resistance
 In 1944, height of WWII, Alexander Fleming was
featured on the cover of Time magazine, with the
subtitle:

“His penicillin will save more lives

than war can spend.”

 About the same time that Fleming’s face appears on

Time magazine, a Stanford researcher publishes
that he has found 5 different strains of Staph that
do not respond to penicillin…The first sign that the
Staph population has responded to penicillin, by
developing resistance.

 This began the “arms race” of development of new

antibiotics in response to antibiotic resistance. A
few examples:
• Streptomycin 1943 (resistance 1948)
• Methicillin 1960 (resistance 1961)
• Clindamycin 1969 (resistance 1970)
• Ampicillin 1961 (resistance 1973)
• Carbenicillin 1964 (resistance 1970)
• Piperacillin 1980 (resistance 1981)
• Linezolid 2000 (resistance 2002) Listen to Radiolab episode
“Staph Retreat”.
From the Virtual Microbiology Classroom on ScienceProfOnline.com
 REVIEW!
See brief SPO Class
Notes articles:
“Early Germ Theory:
Leeuwenhoek,
Semmelweiss
& Snow”

&
“Late Germ Theory:
Contributions
of Pasteur, Lister, Koch,
Domagk & Fleming”

Image: Salmonella, Public Health

From the Virtual Microbiology Classroom on ScienceProfOnline.com Image Library,PHIL #10973
 Early History of Immunology 1749 - 1823

Dr. Edward Jenner and the First Vaccine

 Rural physician aware of farm workers' belief that if

you had cowpox in past, you wouldn’t get smallpox.

 Cowpox caused mild discomfort, aching, a few

pustules, some swelling…symptoms that disappeared in
a few days.

 In contrast, smallpox caused massive disfigurement,

sometimes blindness, and often death.

 Jenner, in the late 1700s, made small incisions or

punctures with cowpox material in arms of human
subjects in order to prevent smallpox.

 At first his peers doubted the safety and efficacy of

his treatment, but eventually the value of the cowpox
inoculum was recognized.

 Jenner’s works are said to have saved more lives than

the efforts of any other person in history.

Images: Dr. Edward Jenner by James Northcote;

From the Virtual Microbiology Classroom on ScienceProfOnline.com Cowpox on Udder Wiki Child with Smallpox, James Hicks, CDC
 Early History of Immunology
Pasteur’s Attenuated Vaccines

 Individuals who recover from an infectious disease

sometimes immune from future attack.

 Prompted Pasteur to try to find a way to prevent fowl

cholera in chickens. 1822 - 1895

 Colleague of Pasteur’s postponed inoculations of cholera

into a group of chickens, a remarkable discovery resulted.

 Inoculation with these neglected cultures made the

chickens immune to fowl cholera.

 The microbes had been weakened or attenuated.

 Pasteur also modified other organisms (anthrax and the

virus causing rabies).

 Ultimately created inoculation procedures … vaccinations.

 REVIEW! See brief SPO Class Notes article: “

Early History of Immunology: Edward Jenner & Louis Past
eur
”
Image: Louis Pasteur, Pierre Lamy Petit, Frenchcirca
From the Virtual Microbiology Classroom on ScienceProfOnline.com 1866, Boy being vaccinated, PHIL #9364.
 Disease, Please! Rabies
 Rabies is an infectious disease caused by a virus.
Rabies virus
 Mortality rate of nearly 100%. looks like little
tiny bullets!
 Description of rabies, from RadioLab “Rodney vs. Death”
Podcast, from WNYC Public Radio (segment time 8:00 to
11:06). See Virtual Microbiology Classroom Main Page for
homework assignment on this podcast.

 Video of rabies in humans.

 Pasteur developed his rabies vaccine

by growing the virus in rabbits, then
drying the infected nerve tissue to
weaken (attenuate) the virus.

 On July 6, 1885, the vaccine was

administered to a 9-year-old boy who
had been attacked by a rabid dog. The
boy did not develop symptoms of
rabies and survived.

 Most rabies cases reported to the CDC

each year occur in wild animals like
raccoons, skunks, bats, and foxes.
Image: Cover of book Rabid, Rabies virus as seen through
electron microscope, PHIL, image #5611;
From the Virtual Microbiology Classroom on ScienceProfOnline.com Portrait of Louis Pasteur in his lab 1885, by Albert Edelfelt
 Confused?
Here are links to fun resources that further
explain aerobic respiration:

• History Microbiology Main Page on the Virtual Microbiology Classroom of Science Prof Online.

• “Got the Time” music video by Anthrax.

• Experiments Testing Spontaneous Generation, animation and quiz.

• The Dress Lodger , a historical novel about cholera, prostitution and body snatching set in Sunderland England
1831.

• Brief History of Microbiology online flashcards from Quizlet.

• Microbe Hunters, a classic book by Paul De Kruif Harvest Books.

• Play “Fling The Teacher”, an interactive “Medicine: Infectious Disease” Quiz in which you get to build a teacher
(victim), then fling them with a trebuchet if you correctly answer the quiz questions.

• History of Microbiology interactive timeline from Microbe World.

• Play Disease Defenders educational video game, Rice University.

From the Virtual Microbiology Classroom on ScienceProfOnline.com
 Are microbes intimidating you?

Do yourself a favor. Use the…

Virtual Microbiology
Classroom (VMC) !
The VMC is full of resources to help you succeed,
including:
• practice test questions
• review questions
• study guides and learning objectives

You can access the VMC by going to the Science Prof Online website
www.ScienceProfOnline.com

Images: Cholera, Vibrio cholerae, Giant Microbes; Prokaryotic cell, Mariana Ruiz