Louis Pasteur ---the master experimenter

N K Srinivasan Ph.D

Louis Pasteur has been an inspiring scientist --chemist and founder of micro-biology--
for thousands of scientifically minded young people and researchers. There are many
facets of this great man that one cannot but admire such a scientist even after more than a
hundred and twenty years. There is also much to learn from his life on the methodologies
of experimentation and analysis of scientific results.

Louis Pasteur was an experimenter of par excellence…Only a few scientists could come
close to him in this respect. He always did simple , elegant experiments which became
‘critical’ in pinning down and establishing the new concepts with conclusive
‘evidences’ or proofs. In my opinion, only a few scientists measure up to him ---they are
Sir Isaac Newton, Michael Faraday and in modern times, Ernest Rutherford and Enrico
Fermi.

The output of Louis Pasteur was also enormous , in several related fields—one field
leading to another. He was not only a ‘microbe hunter’ but also a chemist and an applied
scientist. . In this respect again , the scientists I had mentioned can be comparable to
him.There was a logical progression to his field of research---from fermentation work to
germ theory to immunology.!
In this article , I highlight only some of the achievements and methods of Pasteur ,mainly
with a view to recall them for the present generation of students. In passing, I will also
briefly describe the work of Robert Koch and Paul Ehrlich ,which are related to Pasteur’s
great contributions.

Early Work
Louis Pasteur was born in 1822. He loved to do experiments as a young boy and also
study insects. At the age of 18 or so, Pasteur discovered optical isomerism. He prepared
crystals of sodium ammonium tartarate and observed closely with a magnifying glass.
Observing with a magnifier had been a life –long habit with him! He found two kinds of
crystals, one left-handed and the other right handed---like your left-handed glove and the
right –handed glove. He quickly analyzed the phenomenon behind this difference ---due
to two different molecular structure with asymmetry or isomerism , though both were of
the same composition. What is more, he rightly developed the ‘technique’ for
distinguishing them—one set of crystals turned or rotated the plane of polarization of
light to the left and the other to the right, even after dissolving them in water. What is
more, when he made a solution with equal quantity of the two isomers, there was no
rotation of plane of polarization of light. [This reminds us of the famous experiment by
Sir Isaac Newton; he let in a thin beam of light through a chink in the window and placed
a prism in its path; he obtained the spectrum with rain-bow colors. When he placed
another prism side by side, he found again the white light.]

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Therefore the structure is at the molecular level, the outer shape of the crystal faces
manifesting it.…Thus he established ‘optical isomerism’---a principle widely used in
organic chemistry and applied in saccharimeters and other instruments……This
discovery alone, at that young age, is sufficient to establish Pasteur as a first –rank
scientist in the annals of science.

Critical Experiments

Oftentimes, text books present the final results in a simple, elegant manner. They do not
provide the background leading to the result—how the scientists battled with conflicting
views/hypotheses/theories , often confusing the whole issue or side-tracking it, before
reaching the conclusion regarding the mechanism or concept.
For instance, for a long time, heat was considered a substance which entered or left a
body and not a form of energy created somewhere within the substance. When cannons
were bored, the frictional heat increased the temperature of the cannon, without adding to
any weight! So, critical experiments are required to clinch the issue, nail down a concept
or theory….Pasteur was a superb experimenter in devising such critical experiments. He
would marshal all the known facts, then proceed to design critical experiments which
would eliminate one wrong theory after another. Then he would zero in on the correct
theory.

Let us begin with the case of fermentation of milk and spoiling of wines and beer.
.Pasteur was professor at the University of Lille then. He wanted to help the local
breweries too.
He wanted to prove that the wines soured or fermented due to airborne tiny germs. Kill
those germs already swarming in the wine and also in the surrounding air…then you can
keep wine without getting sour for years.Here is the critical experiment he devised.
He took a glass flask with a long , thin neck. He heated the fermented wine for about 70
deg C to kill the germs in the brew. Then he evacuated the glass flask with a crude pump,
eliminating the air-borne germs and sealed off the long neck with a gas torch….No more
fermentation and he could preserve the brew for months. He would examine the brew for
signs of fermentation…no fermentation at all!…He beautifully demonstrated one more
thing. Break the thin neck of the flask, let in air, and in a few hours, fermentation would
start . Thus the famous ‘swan-neck duct’ experiment was the critical one ---to demolish
the prevalent theory of ‘spontaneous generation’ of germs.
Such critical experiments are the stuff of scientific work---something that a young student
or researcher should learn…Thus Pasteur announced the discovery of air borne-germs
theory in 1857.

From 1857 to 1864---nearly seven years, Pasteur kept a bottle of milk, ‘Pasteurized and
sealed’ without fermentation because he had kept it from ‘ the germs which float in the
air.’
During these years, he also studied the silk-worm disease to find the insect causing it and
also the potato –blight disease.
Note that the process of heating to 70 deg C [or just 65 deg C would do] to kill the germs
is the process now known as “Pasteurization” , announced in 1862..During this

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experimentation, Pasteur had developed a practical means for preserving such food stuff.
This was a milestone in the development of food sciences. We use pasteurized milk
every day even now.!

Spreading a technique

Pasteur was not a scientist doing some isolated work or adding foot note to what others
had done. Pasteur ,after this fermentation work, opened up the germ theory for studying
various human diseases and epidemics. He reasoned that the germs could cause diseases
in animals and humans.

Note that Pasteur was a ‘mere chemist’ and the established medical profession jeered at
him…The doctors said: “What does a mere chemist know about human
diseases.?..”..They engaged him in wordy duels with derogatory remarks in university
halls.Pasteur, like Machael Faraday, had the habit of giving public lectures, with clear
experimental demonstrations, to silence his critics!

Pasteur was undaunted and did work on anthrax and chicken pox at once.
[Joseph Lister, a Glascow Univ professor of Chemistry, was another such breed of
scientists who dare to venture into many fields or even professions. Lister noticed that
when there was an epidemic in cattle at Carlisle, throwing carbolic acid into sewers
controlled the epidemic and the cows recovered….Can carbolic acid work with humans
too.? Inspired by the work of Pasteur, Lister cleaned the wounds of operated patients
with carbolic-acid soaked lints , covered with thin tin foil to exclude air..Lister next
treated the hospital air with hand-operated carbolic acid spray. This technique
revolutionized surgery and brought down death in hospitals due to infections.]

Pasteur worked on anthrax disease [“foot and mouth disease”] among cattle and also
other infections. He wanted to isolate the germ responsible for this disease.

At that time, there was bitter warfare between France and Germany. But science knows
no national boundaries. A village general practitioner [GP], Dr Robert Koch [1843-1910]
with a simple optical microscope in his tiny clinic, found the anthrax bacilli under the
microscope, and cultured it in 1876. He also developed the technique of staining the
organism with chemical dyes with the help of his friend Paul Ehrlich[1854-1915] another
microbe hunter. Koch rightly proved that these bacilli produced spores in the dead
animal tissues which spread the disease by depositing endospores in loose soil. Instead
of using a broth to culture these bacteria, Koch used ‘synthetic solutions’ with gelatin
and nutrients to grow them---Thus isolate the bacteria, grow them in synthetic culture
and observe under the microscope with dye-staining technique; [using aniline dyes at that
time.] Thus the branch of bacteriology was born with this trio—Louis Pasteur, Robert
Koch and Paul Ehrlich. [1]

Pasteur always promoted exchange of information and communication between

scientific workers. He encouraged Lister in his work in Scotland. He was communicating

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with Koch who was not a university professor , but a private investigator and Paul
Ehrlich in Berlin among others.

Fighting diseases and immunology

The eventful year of 1882 dawned. Pasteur and Koch both announced the isolation of the
bacteria for anthrax disease which was killing cattle in hundreds. What a triumph.!
Koch also isolated the great killer, the tubercle bacilli , responsible for tuberculosis in
humans.
It is one thing to find the micro-organism or microbe for the cause of a disease, but
another to find a cure or preventive method.… How to protect oneself against these
microbe-invaders?..They are always there swarming around us. Pasteur , inspired by
early work on small pox [by Edward Jenner], theorised that the degraded forms of micro-
organisms can create immunity in the body against the same disease…Now we know that
they produce anti-bodies in us. This principle, Pasteur tested with graduated doses of the
bacilli. He tested in animals and then in humans.

Thus Pasteur established the branch of study of micro-biology and founded

immunology…Swift results followed. Dr Robert Koch visited India and isolated the
bacilli for cholera which swarmed in soiled clothes and contaminated water ,with his
simple microscope and staining technique. In 1879, gonorrhoea bacillus was isolated; in
1881,streptococus;in 1883,diphtheria;in 1884 typhoid and tetanus; in 1905 ,syphilis
bacillus. Koch, Ehrlich and their students found the bacilli causing other diseases as
well---meningitis, leprosy, bubonic plaque and elephantiasis…By the first decade in 20th
century, almost all bacterial diseases have been identified with their specific bacteria.
Thus Pasteur ushered in the era of microbe hunters and their victory over several
bacteria-induced serious diseases. [2]

Vaccines for many diseases

Pasteur was not a sceintist who would rest on his laurels. He was sixty-two years old
now. That was a pretty senior year in those days when average life span in Europe was
around 50 years. He had his first stroke in 1868 and was half-paralysed. He walked with
great difficulty, limping always.
His devoted wife of several decades, Marie Laurent, daughter of the rector in the Univ
at Strassborg, helped him in all daily work. Her main role was to record the results of
experiments systematically in a log book in the evening hours, while Pasteur dictated..
Pasteur was meticulous in this task. His lab notebooks themselves served to clarify
problems in a well-ordered manner.[Incidentally, both Michael Faraday and Thomas
Edison had similar habits of writing down their lab work systematically---something that
our young science students should learn---record your observations, both positive and
negative then and there---or at least that day itself; or else you may forget many subtle
observations!!]

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Now Pasteur prepared himself to attack the problem of infection from bite of mad dogs
—hydrophobia or rabies. He had seen many young persons suffering from that disease in
his village. Could he find a vaccine for this? The next three years he worked hard and
found the vaccine which is used through out the world even today. There are poignant
stories of his testing this vaccine with small boys. In the first instance, injecting a boy,
Joseph Meister , nine year old, Pasteur was taking a bold , risky step. Pasteur was not a
medical doctor. How could he inject an infected boy? He consulted with his colleagues
and then decided to go ahead. The boy recovered and soon the medical profession
accepted his vaccine.[3]
Pasteur , after having multiple strokes, died peacefully in 1895 at the age of 72.
A famous quotation of Pasteur is this: “success comes to those with a prepared mind.”

His religion
His religion was centered around serving the humanity through science. . He was a
devout Catholic throughout his life. Historians have debated whether he attended mass
regularly and all that. He had great reverence of Catholic saints and he died listening to
the story of St Vincent de Paul whom he admired. He was buried in Notre Dame
Cathedral in Paris.

References

1 Paul de Krieuf ---Microbe Hunters—Dover Pub

2 Axel Munthe—The story of San Michele
3 Rene J Dubois—Louis Pasteur--- ( a definitive biography)
4 A C Quiller-couch – Roll Call of Honor—Clarenden Press, London
5 James Burke—The day the universe changed – Little,Brown and company,London
6 C Barlow—Robert Koch—Heron Books

Notes
1 Nobel prizes were instituted in 1900 and thus Pasteur could not receive the award. But
Koch received the Nobel prize for medicine in 1905 and Paul Erhlich received it in the
year 1909.
2 It may be noted that the average life span of humans increased substantially due these
victories, supported by public health measures. The next major step was with the
discovery of antibiotics in 1940’s.
3 Pasteur Institutes were founded in many major cities to prepare vaccines in large
quantities following his methods in his own lifetime.

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About the Author

The author , Nenmeli K Srinivasan ,studied chemistry at University of Madras, India and
then studied metallurgical engineering at Indian Institute of Science , Bangalore. He
obtained his doctorate from Columbia University , New York in 1972, working on
nuclear materials with US Atomic Energy Commission fellowship. He is now retired
after active service and writes articles for science journals and magazines. He spends his
time between Palo Alto,California and Bangalore,India.
Contact: nksrinivasan@hotmail.com
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