ASSIGNMENT

Topic: Historical foundation and development

of Microbiology.

Submitted by: Submitted to:

Ameekha Zakeer Dr. Nisha P Thirumeni
S1 MSc. Microbiology Bioscience Department
MES College, Marampally
29/11/2021
CONTENT:
1) Introduction
2) Important discoveries in the field of Microbiology
3) Antony van Leeuwenhoek
4) Edward Jenner
5) The theory of Spontaneous generation
6) Louis Pasteur
7) Joseph Lister
8) Robert Koch
9) Alexander Fleming
10) Conclusion
11) Reference
INTRODUCTION
The study of microbes is known as Microbiology (Micros =small; logos = Study).
Even though man has been experiencing the influence of microbes since time
immemorial, the scientific study of microbes is comparatively recent. The science
of microbiology was unknown before the discovery of that most important tool of
microbiologist – the microscope.

From the first century BC it was believed that disease are caused by invisible beings.
During the 13th Century, Roger Bacon (1220-1292), suggested that disease are
caused by invisible beings. The opinion was supported by Girolamo Francestoro
Verona (1483-1553) and Anton von Plenciz in 1762.Earlier than Anton von Plenciz,
a monk – Anthanasius Kirchers – related the decay of bodies,meat, milk etc., in
invisible worms.

The most important discovery that may be called as the founding step of the
science of microbiology was that Antony van Leeuwenhoek, a Dutch scientist who
lived in Holland from (1632-1723). With his crude optical device, which is the
forerunner of the modern day microscope, he observed tiny microbes in a drop of
pond water which he called animalcules. These are now regarded as bacteria. Since
then started the glorious era of microbiology.
In the history of microbiology three specific phases may be recognized.

These are:

(1) Discovery phase

(2) Transition phase, and
(3) The golden age

During the discovery phase, the microbes were discovered and attention was
devoted to mainly observe and describe the microbes. The names of Marcello
Malphigi and Antony van Leeuwenhoek are important during this period.

The transition period in the development of microbiology coincided with a great

controversy regarding the orgin of microbes. The controversy arose as to the
opinion of some, that life arises spontaneous! From a non- living source (abigonetic
orgin of life), as one can see the sudden appearance of tiny maggots, worms and
flies as if from nowhere on any decaying organic matter. The experiments of
Fransisco Rhedi, John Needham, Spallanzani and Nicolas Apperte are worth
mentioning.

The third phase in the history of microbiology, which may be reffered to as the
golden age, began in 1860 and continued up to 1910 when many important
contributions were made. The most significant contribution during this period
came from Louis Pasteur and Robert Koch.
Important Discoveries In The Field Of
Microbiology:

Name of the Scientist Year Discovery

1.Anthanasius Kircher (1601-1680) Invisible worms cause

diseases.

2.Girolamo Francastoro (1483-1553) Invisible beings cause

diseases.

3.Francesco Rheedi (1626-1697) Disproved the theory of

spontaneous generation.

4.Antony van (1632-1723) Pioneer in the discovery

Leeuwenhoek of microbes.

5.John Needham (1713-1781) Conducted experiments

which seemed to prove
the theory of
spontaneous generation.

6.Lazaro Spallanzani (1729-1799) Experimentally disproved

the theory of
spontaneous generation.
7.Edward Jenner (1749-1823) Discovered vaccine
against small pox

8.Justus von Leibeg (1803-1873) Discovered chemical

basis of fermentation.

9.Jacob Henle (1809-1885) Established the

fundamental principles of
the germ theory of
diseases.

10.Louis Pateur (1822-1895) Discovered fermentation

by living cells;
conclusively disproved
the theory of
spontaneous generation;
developed rabies
vaccine.

11.Joseph Lister (1827-1912) Introduced antiseptic

method in surgery.

12.T.J Burill (1839-1916) Discovered bacterial

plant diseases.

13.John Tyndall (1839-1916) Discovered fractional

killing of bacterial spores.
14.Robert Koch (1843-1910) Proposed Koch’s
postulates for germ
based diseases; identified
tuberculosis causing
bacilli

15.Paul Ehrlich (1854-1915) Developed the principles

of chemotherapy to cure
diseases.

16.Elie Metchnikoff (1845-1916) Discovered the

phenomenon of
phagocytosis

17.Hans Christian Gram (1853-1933) Discovered differential

staining of bacteria using
gentian violet dye

18.N.Winogradsky (1856-1934) Discovered N2 fixing

bacteria in the soil.

19.William Welch (1850-1934) Discovered the relation

of anaerobic bacteria to
gangrene.
20.Walter Reed (1851-1902) Discovered the
transmission of yellow
fever by mosquitoes.

21.Theobald Smith (1909) Discovered rickettisiae

22.Theobald Smith (1859-1934) Discovered that Texas

fever is transmitted by
ticks.

23.Ronald Ross (1897) Discovered the sexual

cycle of malaria parasite
in mosquito.

24.Jules Bordet (1906) Isolated the whooping

cough bacillus.

25.David Bruce Discovered that sleeping

sickness disease causing
microbes is transmitted
by tse tse fly.

26.George Gafky Isolated typhoid bacillus.

27.Shiba saburo Kitasato Isolated teanus bacillus.

28.Emil von Behring Development of anti

diphtheria toxin.
Antony van Leeuwenhoek
Father of microbiology – Antony van Leeuwenhoek

Leeuwenhoek was a merchant by profession but he ground lenses and made

microscopes as a hobby. He had no formal University education but had a keen
mind. During his life time Leeuwenhoek constructed more than 200 microscopes.
The 'microscopes' of Leeuwenhoek could magnify objects about 200-300 times.

With his microscope, Leeuwenhoek observed a variety of things mainly out if

curiosity. Hair fibres, plant structures, crystals, insect, insect's eye, a variety of fluids
such as pond water, blood etc, scrapings from his own teeth and transparent tail
fins of fish were observed by him.

For some 50 years Leeuwenhoek kept meticulous records and made accurate
drawings of his observations. He examined blood and discovered some tiny
microbes which are nothing but erythrocytes. He examined yeasts and found that
they were made up of tiny round particles.

He observed under his microscope minute moving objects which he called

animalcules (small animals) which we now know as protozoa, fungi, and bacteria.
He communicated his findings to the Royal Society of London where his
observations were translated into English and published in the Proceedings of the
Royal Society as a series of letters.

EDWARD JENNER:

❖ Father of Vaccination
❖ Father of Immunology

Edward Jenner, born in mid-18th century England, would eventually become one of
the most famous scientists in medical history and the so-named “Father of
Immunology.”He was a British physician and scientist who pioneered the concept
of vaccines including creating the smallpox vaccine, the world’s first ever vaccine.
The terms vaccine and vaccination are derived from Variolae vaccinae (‘smallpox of
the cow’), the term devised by Jenner to denote cowpox.

In May 1796 a dairymaid, Sarah Nelmes, consulted Jenner about a rash on her hand.
He diagnosed cowpox rather than smallpox and Sarah confirmed that one of her
cows, a Gloucester cow called Blossom, had recently had cowpox. Edward Jenner
realised that this was his opportunity to test the protective properties of cowpox
by giving it to someone who had not yet suffered smallpox.

He chose James Phipps, the eight-year old son of his gardener. On 14th May he
made a few scratches on one of James' arms and rubbed into them some material
from one of the pocks on Sarah's hand. A few days later James became mildly ill
with cowpox but was well again a week later. So Jenner knew that cowpox could
pass from person to person as well as from cow to person. The next step was to
test whether the cowpox would now protect James from smallpox. On 1st July
Jenner variolated the boy. As Jenner anticipated, and undoubtedly to his great
relief, James did not develop smallpox, either on this occasion or on the many
subsequent ones when his immunity was tested again. This may be truly regarded
as one of the greatest achievements ever made by man in his quest for conquering
of disease and is also heralded as a significant milestone in the history of
microbiology.

(NB: Small pox virus, however has been completely eradicated from the earth and
immunization against it is no longer necessary.)

The Theory Of Spontaneous Generation:

The doctrine of 'spontaneous generation’ or 'abiogenesis' was mainly the belief in
the spontaneous formation of living beings from nonliving matter. The theory, that
new forms of life can arise spontaneously from a non-living source, is called the
Theory of Spontaneous generation or the abiogenetic orgin of life. On the other
hand, the theory that states that life can come only from pre-existing life is called
the biogenetic orgin of life.

However, one of van Helmont’s contemporaries, Italian physician Francesco Redi

(1626–1697), performed an experiment in 1668 that was one of the first to refute
the idea that maggots (the larvae of flies) spontaneously generate on meat left out
in the open air. He predicted that preventing flies from having direct contact with
the meat would also prevent the appearance of maggots. Redi left meat in each of
six containers (Figure 1). Two were open to the air, two were covered with gauze,
and two were tightly sealed. His hypothesis was supported when maggots
developed in the uncovered jars, but no maggots appeared in either the gauze-
covered or the tightly sealed jars. He concluded that maggots could only form when
flies were allowed to lay eggs in the meat, and that the maggots were the offspring
of flies, not the product of spontaneous generation.

Figure 1. Francesco Redi’s experimental setup consisted of an open container, a container sealed with a cork top,
and a container covered in mesh that let in air but not flies. Maggots only appeared on the meat in the open
container. However, maggots were also found on the gauze of the gauze-covered container.
In 1745, John Needham (1713–1781) published a report of his own experiments, in
which he briefly boiled broth infused with plant or animal matter, hoping to kill all
preexisting microbes.He then sealed the flasks. After a few days, Needham
observed that the broth had become cloudy and a single drop contained numerous
microscopic creatures. He argued that the new microbes must have arisen
spontaneously. In reality, however, he likely did not boil the broth enough to kill all
preexisting microbes.

Lazzaro Spallanzani (1729–1799) did not agree with Needham’s conclusions,

however, and performed hundreds of carefully executed experiments using heated
broth.As in Needham’s experiment, broth in sealed jars and unsealed jars was
infused with plant and animal matter. Spallanzani’s results contradicted the
findings of Needham: Heated but sealed flasks remained clear, without any signs
of spontaneous growth, unless the flasks were subsequently opened to the air. This
suggested that microbes were introduced into these flasks from the air. In response
to Spallanzani’s findings, Needham argued that life originates from a “life force”
that was destroyed during Spallanzani’s extended boiling. Any subsequent sealing
of the flasks then prevented new life force from entering and causing spontaneous
generation .

Figure 2. (a) Francesco Redi, who demonstrated that maggots were the offspring of flies, not products of spontaneous generation. (b) John
Needham, who argued that microbes arose spontaneously in broth from a “life force.” (c) Lazzaro Spallanzani, whose experiments with broth
aimed to disprove those of Needham.
LOUIS PASTEUR:
Pasteur's contribution to the development of microbiology are enormous, and he
may be rightly called the Father of Modern Microbiology. His main contributions
are :-

1.Tartaric acid, an organic compound, is formed by two types of crystals which

could be separated microscopically.

2.Discovery of microbial fermentation : Pasteur turned his attention to the

phenomenon of fermentation because France was a leading wine exporter.
Fermentation was thought to be a chemical process without the involvement of
living organisms. The French wine industry at that time faced the problem of wine
getting sour or ropy due to spoilage. Pasteur took up the problem of souring of
wine and beer. Pasteur found that acid wines, ropy wines and sour beer were due
to the growth of undesirable organisms, while the desirable organism gave rise to
a good product. After a great deal of work, Pasteur showed that wine did not spoil
if it were held for a few minutes at 50- 60°C.

3.Disproving the theory of spontaneous generation : Pasteur made a series of

flasks with long, twisted necks (“swan-neck” flasks), in which he boiled broth to
sterilize it (Figure 3). His design allowed air inside the flasks to be exchanged with
air from the outside, but prevented the introduction of any airborne
microorganisms, which would get caught in the twists and bends of the flasks’
necks. If a life force besides the airborne microorganisms were responsible for
microbial growth within the sterilized flasks, it would have access to the broth,
whereas the microorganisms would not. He correctly predicted that sterilized broth
in his swan-neck flasks would remain sterile as long as the swan necks remained
intact. However, should the necks be broken, microorganisms would be
introduced, contaminating the flasks and allowing microbial growth within the
broth.

Figure 3. (a) French scientist Louis Pasteur, who definitively refuted the long-disputed theory of spontaneous generation. (b) The unique swan-
neck feature of the flasks used in Pasteur’s experiment allowed air to enter the flask but prevented the entry of bacterial and fungal spores. (c)
Pasteur’s experiment consisted of two parts. In the first part, the broth in the flask was boiled to sterilize it. When this broth was cooled, it
remained free of contamination. In the second part of the experiment, the flask was boiled and then the neck was broken off. The broth in this
flask became contaminated.

4.Discovery of aerobic and anaerobic microbes: Microbes they require oxygen to

grow are called aerobes. Since animals require oxygen for growth, scientists in the
middle of the 19th century, assumed that microbes also require oxygen. However,
Pasteur’s study of the process of butyric acid fermentation, reveled that microbes
can survive in the absence of oxygen. Microscopic observation of the fermented
product showed actively moving microbes. The mobility of the microbes however
stoped when they were exposed to air. Pasteur also observed that if the broth is
aerated, fermentation would stop. He coined the term anaerobic to refer to
organisms that do not require oxygen.

5. Work silkworm disease: In 1865,Pasteur demonstrated that the silkworm

diseases called ‘pebrine' was caused by a microscopic germ – a protozoan, and
showed that the infection could be controlled by choosing worms free from the
parasite for breeding purpose.

6. Work on anthrax and rabies disease: Pasteur demonstrated that the anthrax
disease in sheep are were caused by microbial rods (bacteria) and cultured them in
sterilized yeast water and showed that the inoculation of culture would cause the
disease in healthy animals. He also discovered the principle of protective
inoculation or vaccination against the disease. He showed that inoculation of
weekend microbes to animals would provide them with immunity and they would
not contract the disease- thus was discovered the anthrax vaccine.

Pasteur successfully applied the principle of protective vaccination to another

dangerous human disease – rabies (hydrophobia), caused due to mad dog bite.
Pasteur, however, could not observe the casual agent ( virus) under his microscope.
But, he could develop a vaccine of weakened organisms (viruses) that provided
immunity against this near fatal disease.

7. Pasteurization: The process of partial killing of microbes called pasteurization

that is now widely employed in dairies was originally devised by Pasteur to prevent
spoilage of wine and beer.
JOSEPH LISTER :
Lister's contribution to surgery are many, but his major contribution has been in
preventing wound infection after surgery by following antiseptic methods. Lister,
as a surgeon, was moved by the post – operative complications in patients due to
wound infection and set upon to study the reasons for it. He published his research
papers on this aspect in 1867.He observed a similarity between the wound
inflammation and putrefaction and fermentation which Pasteur had already shown
to be due to microorganisms. This lead him to the conclusion that microbial growth
may be the cause of wound infection. He was sure that if he would protect the
wound with dressing that killed the microbes, inflammation would never occur.
This was the basis of antiseptic technique.

Lister chose carbolic acid (a well known preservative), as the microbiological agent.
He applied carbolic acid soaked dressing to cover the wound of compound fractures
which usually caused (often fatal) inflammation. The results of the application of
carbolic acid dressing were dramatic; wounds healed in double – quick lime. Lister
applied this technique to many fastering wounds and obtained similar results.
Finally, Lister developed a routine for the use of carbolic acid during operations
which came to be known as, Lister's antiseptic system.

ROBERT KOCH:
That germs were responsible for diseases was suspected even before the
experiments of Pasteur. However, the definite knowledge that germs were the real
causes of several ailments was established and the concept ‘germ theory of
disease' was conceived only with the work of Pasteur. Robert Koch, a German
scientist, firmly established that germs were the causes and not the end products
of diseases. Koch was a physician who later became the professor of Hygiene and
Director of Institute of Infective Diseases at Berlin. He discovered bacilli in the blood
of cattle that had died of anthrax. He grew these bacteria in the cultures examined
them microscopically and injected them into healthy animals. The infected animals
developed anthrax symptoms and from the infected animals he reisolated similar
bacilli. These series of experiments led to the establishment of 'Koch’s postulates’
which provided the method to identify the real causative agent of a disease. Koch's
Postulates' are :

1) A specific orgamism should be found constantly in association with a

disease.
2) The organism should be isolated and grown in a pure culture in the
laboratory.
3) The pure culture when inoculated into a healthy susceptible animal should
produce symptoms of the same disease.
4) From the inoculated animal it should be possible to recover the organism
with the same characteristics as the original isolate.

Koch is also remembered for his contribution to the development of pure culture
techniques. For culturing the microorganisms Koch originally used gelatin to
prepare transparent jelly on flat pieces of glass. However, gelatin melted at 25°C
and it was not possible to use this medium at high temperatures. The replacement
of gelatin came rather accidentally. One of Koch's students was W. Hesse. Jesse’s
wife Fannie suggested in 1881 that agar – agar derived from sea weed (Gelidium
sp.) was being used for making jellies in the kitchen and could be used for the
preparation of media. Koch experimented on the idea and found that agar-agar was
most suitable for this purpose as it had a melting point above 100°C and a jelling
point of 45°C which is ideally suited for several laboratory purposes. Besides, agar-
agar is totally inert with no nutritive value and undigestable for most organisms. At
1.5 per cent it forms a perfect transparent gel. Even after more than a century of
use, agar-agar has not yet been displaced by a better substance!

Robert Koch also developed techniques of staining smears of bacteria with aniline
dyes for better microscopic observation. He discovered the Spirillum responsible
for cholera in 1883 and the agent of tuberculosis Mycobacterium Tuberculosis in
1882.His contributions to microbiology provided tremendous advances by the turn
of the 19th century.

ALEXANDER FLEMING:
Sir Alexander Fleming (6 August 1881 – 11 March 1955) was a Scottish physician
and microbiologist, best known for discovering the world’s first broadly effective
antibiotic substance, which he named penicillin. His discovery in 1928 of what was
later named benzylpenicillin (or penicillin G) from the mould Penicillium rubens is
described as the “single greatest victory ever achieved over disease.”He also
discovered the enzyme lysozyme from his nasal discharge in 1922, and along with
it a bacterium he named Micrococcus Lysodeikticus, later renamed Micrococcus
luteus.

By 1927, Fleming had been investigating the properties of staphylococci. He was

already well known from his earlier work, and had developed a reputation as a
brilliant researcher. In 1928, he studied the variation of Staphylococcus aureus
grown under natural condition, after the work of Joseph Warwick Bigger, who
discovered that the bacterium could grow into a variety of types (strains). On 3
September 1928, Fleming returned to his laboratory having spent a holiday with his
family at Suffolk. Before leaving for his holiday, he inoculated staphylococci on
culture plates and left them on a bench in a corner of his laboratory.On his return,
Fleming noticed that one culture was contaminated with a fungus, and that the
colonies of staphylococci immediately surrounding the fungus had been destroyed,
whereas other staphylococci colonies farther away were normal, famously
remarking “That’s funny”.Fleming showed the contaminated culture to his former
assistant Merlin Pryce, who reminded him, “That’s how you discovered
lysozyme.”He identified the mould as being from the genus Penicillium. He
suspected it to be P. chrysogenum, but a colleague Charles J. La Touche identified
it as P. rubrum. (It was later corrected as P. notatum and then officially accepted as
P. chrysogenum; but finally in 2011, it was resolved as P. rubens.)

Fleming grew the mould in a pure culture and found that the culture broth
contained an antibacterial substance. He investigated its anti-bacterial effect on
many organisms, and noticed that it affected bacteria such as staphylococci and
many other Gram-positive pathogens that cause scarlet fever, pneumonia,
meningitis and diphtheria, but not typhoid fever or paratyphoid fever, which are
caused by Gram-negative bacteria, for which he was seeking a cure at the time. It
also affected Neisseria gonorrhoeae, which causes gonorrhoea, although this
bacterium is Gram-negative. After some months of calling it "mould juice" or "the
inhibitor", he gave the name penicillin on 7 March 1929 for the antibacterial
substance present in the mould.
CONCLUSION:
Perhaps no other developments have had such a major impact on the health and
welfare of humankind than those comprising the history of microbiology and
immunology. Over the last 100 years, the mortality burden of infectious diseases
has decreased substantially and the average lifespan has increased by over 30 years
due to advances in public health, sanitation, vaccines, and anti-infective
chemotherapy – all deriving from the sciences of microbiology and immunology
(Centers for Disease Control 1999). Future advances are anticipated when the
genomics era in which we now live and the promise of systems biology and
personalized medicine are fully realized in the next few decades. A remarkable
story of directed inquiry into the fundamental nature of microbes and immune
defenses preceded many of the current advances in medicine. Much work remains
before the benefits of these discoveries can be applied equally worldwide.
REFERENCE:
Books:

1) General Microbiology, Second edition, S. B Sullia, S. Shantharam

2) General Microbiology, Seventh Edition, Hans G. Schlegel

WEBSITE:

1) https://courses.lumenlearning.com/microbiology/chapter/spontaneous-
generation/
2) https://www.frontiersin.org/articles/10.3389/fimmu.2012.00068/full