MICROBIOLOGY (BT 13102)

Contributions of Louis Pasteur : French biologist, microbiologist, and chemist

(1822-1895)

A. He began his career as professor in chemistry at university of Lille, France. Principal industry i.e.
Wine and Beer was facing a problem

• He studied and found that fermentation of fruits and grains which resulted in production of alcohol
was brought about by microbes. He examined many batches and found microbes of different types

• In good lot one type predominated and in poor products another kind was present.

• By proper selection of microbe, consistent good and uniform product can be achieved.

• He suggested that undesirable type of micro organism can be removed by heating which is not
enough to destroy the flavour of fruit juice but enough to destroy a very high % of microbial
population.

• He postulated that holding juices at 62.80C (1450 F) for 30 minute about good results –
Pasteurization – Dairy industry.

B. After solving fermentation problem, French Government requested him to investigate pebrine
silkworm disease that ruined French silk industry. For many years he remained busy with this problem
and ultimately he isolated parasite causing the disease. He advocated that silkworm farmers could
eliminate the disease by using only disease free caterpillars for breeding stock.

C. CHICKEN CHOLERA EXPERIMENT

 He isolated bacterium responsible for chicken cholera and grew it in pure culture. To prove that he
really had isolated organism responsible for chicken cholera he organised a public demonstration.
He inoculated healthy chicken with pure culture but to his disbelief the chicken didn’t die.

 He reviewed each step and found that he had accidently used cultures several week old instead of
fresh one grown specially for demonstration. Some weeks later he again repeated experiment using
two group of chicken.
 One group has been inoculated at first demonstration with old culture and second had not been
previously exposed with group received bacteria from fresh young cultures.

 This time the chicken of second group got sick and died and those in first group remained healthy.

 He explained that in some way bacteria could lose its ability to produce disease (Virulence) after
standing and growing old. But these attenuated culture (decreased Virulence) still retained the
capacity for stimulating to produce substance i.e. antibiotic that protect against subsequent
exposure to virulent organism.

 This demonstration explained the principle involved in Edward Jenner use of small pox virus. In
1798 to immunise people against small pox.

 Pasteur (1822-1895) applied same principle to prevention of anthrax and it again worked and he
called attenuated cultured as Vaccines – (Latin word – Vacca – cow)

D. He was asked to work on Rabies – disease affecting human beings. This disease caused by bite of
dog, cat and other animals. Joseph Mister was bitten by a dog and as the disease was fatal his
family took a chance.

 As rabies virus was too minute to be seen by microscope and never grown in lab culture as it was
not bacterium. The disease was produced in rabbits by inoculating them with saliva from mad dogs.
Then brain and spinal cord if infected rabbit was removed, dried for several days, pulverized and
mixed with glycerine. Injecting this mixture into dogs protected them against rabies but vaccinating
was different from treating a sick boy. He carried the trial which took several weeks (1860-1910) and
Joseph didn’t die.

 Contributions of Robert Koch (1843-1910)

A. He discovered the typical bacilli with squarish ends in the blood of cattle that had died of anthrax.

• He grew these bacteria in culture in his lab and injected them to other animal to see if these animals
become infected and developed clinical symptoms of anthrax.

• From those he had originally found in sheep that had dead of anthrax.

• This was first time that bacteria had been proved to be the cause of animal disease.
• These observations led to establishment of Koch Postulates which provided guidelines to identify
causative agent of disease.

B. Koch’s Postulates/Germ Theory of Disease

1. A specific organism is responsible for causing a particular disease.

2. The organism can be isolated and grow in pure culture in lab.

3. The pure culture will produce the disease when inoculated into susceptible animal.

4. It is possible to recover the organism in pure culture from experimentally infected animal.

C. Koch was concerned with need to develop a technique for the isolation of micro organism in pure
culture in order to establish causative agent of disease.

 He experimented with slices of potato as a solid surface upon which colonies can grow. It proved
ineffective. Then he tried gelatine as solidifying agent.

 It had the advantage of transparent gel but it had disadvantage of becoming liquid above 25 0C
which is opt temp for growth of human disease produce bacteria.

 The solution was provided by Fannie Hesse in 1883 who spent her time working in lab of her
husband Walther Hesse.

 She suggested him to use agar (polysaccharide of algal origin) a substitute of gelatine and he
ultimately reported it to Koch.

Melting point – 1000C ; Solidifies – 450 C

This feature make it possible to incubate the inoculated media at any desired temperature and still
media remained solid.

D. Koch found that smearing bacteria on a glass slide and adding certain dyes to them, the individual
cells can be seen more clearly with microscope.

 He added gelatine and other solidifying materials such as agar to media in order to obtain
isolated growth of organism known as colonies.

E. R. Koch: Anthrax , Tuberculosis, Cholera as well as concept of Pure Culture

 Bacillus antracis, Mycobacterium tuberculosis, Vibrio cholera

Other Contributors

• Richard Petri (Kochi assistant) developed petridish for solid culture media and their made possible
the isolation of pure culture that contained only one type of bacterium.

• The discovery of viruses and their role in disease was made possible when Charles Chamberland
(1851-1908). Pastures also create constructed a porcelain bacterial filter in 1884 and first
pathogen to be studied was TMV.

• Walter Reed – Reported transmission of yellow fever of mosquito.

• Wassermann – Introduced complement fixation test

Medical Microbiology

 Edwin Klebs in 1883 and Frederick Loeffler in 1884 discovered diphtheria bacillus and showed
that it produced its toxin poisons in lab flask.

 Emil von Behring and Shibasaburo Kitasato devised a method of producing immunity to
infections caused by these organisms by injecting their toxins in to animals so that an antitoxin
(substance that neutralizes toxin) would develop.

 Kitasato cultivated Clostridium tetni (tetnus/lock jaw) and with Behring made antitoxin for the
prevention and treatment of the disease. He was awarded Noble prize – Serum therapy.

 Salmon and Theobald Smith demonstrated that immunity to many infections could be produced
by inoculation with killed cultures of mos.

 Elite Metchnikoff (Russian) showed how leucocytes (WBC) could ingest disease producing
bacteria in the body. He called these special defenders against infection as phagocytes and the
process phagocytises. He postulated that phagocytes were the body’s first and most important lie
of defence against infection.

 One of the student of Koch – Paul Ehrlich had different concept of how the body destroys bacteria
and he explained immunity on the basis of certain soluble substances in the blood. He also found
that 606th compound tested i.e. organic chemical compound containing arsenic can destroy
syphilis microbe in body. This was the first chemotherapy substance scientifically discovered and
evaluated.
 Joseph Lister (English Surgeon) was trying to combat the microbes that caused post operative
and wound infection. Disinfection were not known and only carbolic acid would kill bacteria. Lister
used a dilute solution of this acid to soak surgical dressing. Wounds protected in this way didn’t
become infected and healing took place rapidly. This antiseptic surgical practice established the
principles of present day aseptic technique.

Agricultural, Industrial and Environment Microbiology

 In late 1800 Sergi Winogradsky showed importance of bacteria in taking nitrogen from
atmosphere, combining it with other elements and making it available plant food. (N 2 fixing bacteria
in soil)

 Hellriegel and Wilfarth in 1888 showed mutually beneficial relation between bacteria and leg
plants.

 Martines Willem Beijernick (Dutch Microbiologist) found the free living nitrogen bacterium
Azotobacter and described its usefulness in promoting soil fertility.

 1904 – Hiltner – Rhizosphere

 Emil Christian Hansen opened the way to study of industrial fermentations. He developed pure
culture study of yeast and bacteria used in vinegar manufacture and pure cultures were known as
starters.

 Thomas Jonathan Burrill found that in peers a disease called fire blight was caused by
bacterium. This discovery opened a new field plant pathology.

Some of the early microbiologist chose to investigate the ecological role of micro organisms. They
studied microbial involvement in C, N and S cycles taking place in soil and aquatic habitats. Prominent
are Sergei Winogradsky and M. Beijernick.

• Winogradsky – He made many contribution to soil microbiology. He discovered soil bacteria that
could oxides Fe, S and NH4 to obtain energy. He also postulated that many bacteria could
incorporate CO2 into organic matter much like photosynthesis organism do. He also isolated
anaerobic N fixing soil bacteria and studied the decomposition of cellulose.
• M.Beijernck - He made several contribution of microbial ecology. He isolated anaerobic N fixing
Azotobacter, root module bacterium capable of fixing N2 – (Rhizobium) and sulphate reducing
bacteria.

• Beijernick and Winogradsky developed the enrichment culture technique

Branches and Scope:

Medical Causative agents of disease Diagnostic Procedure – identification

Preventive measure

Food Microbiology Preservation, Preparation, Food borne disease, prevention

Agriculture Soil fertility, biofertilizers, biopesticides etc..Eco restoration etc.

Industrial Vaccine, antibiotics, beer, wine, chemicals, protein and hormones by

genetically eng. Microbe

Geochemical Coal, minerals, gas, recovery of mineral ...

Exomicrobiology Outer space exploration

Aeromcrobiology Air, contamination and spoilage dissemination of disease.

Aquatic Water purification,

Microbiological examination, biological degradation etc...