Lesson 1.

HISTORICAL PERSPECTIVE OF MICROBIOLOGY

Module 1. History and scope of microbiology

Lesson 1
HISTORICAL PERSPECTIVE OF MICROBIOLOGY

1.1 Introduction

The history of microbiology can be traced to ‘First Epidemics’ on earth, which

affected Caveman and were probably waterborne. People had no real
understanding of why disease occurred. As the civilization progressed, people
started clustering into cities. They increasingly shared communal water, handled
unwashed food, and stepped in excrement from casual discharge. The crowding
increased and spread water-borne, insect-borne and skin-to-skin infectious
diseases. Yet there was no general understanding of why disease occurred. By the
13th century fear of the diseased took a drastic turn in the formation of small leper
colonies intended to isolate people carrying the devastating disease caused
by Mycobacterium leprae. In 1348, a mass epidemic caused by a single
organism, Yersinia pestis, wiped out nearly one third of Europe's population. The
Plague spread rapidly in the unsanitary conditions of the Middle Ages, leaving
Medieval Europeans defenseless against its devastation. Entire towns succumbed
to the disease, leaving the living to dispose of thousands of contaminated corpses.
Perhaps the deadliest disease in history, the plague or ‘Black Death’, claimed over
20 million lives and contributed to the fall of empires.

1.1.1 Bubonic plague

The social and political dislocations as a result of Bubonic plague were immense,
and the resulting terror was particularly intense because the cause of the disaster
was unknown. Not until 500 years later, in 1890, did microbiologists identify the
causative organism, Yersinia pestis, carried by infected fleas. Infection spreads
among the rats as fleas bite them. But rats have little resistance to plague and
usually die. When rats become scarce, the fleas move to humans and a plague
epidemic is under way. Although rare today, Bubonic plague still occurs in parts of
the world.

1.1.2 Potato blight

The Irish famine Potato blight, a disease of plants, rather than humans, caused by a
fungus rather than a bacterium, had even greater impact than the plague. Potato
blight was responsible for the great Irish famine of the 1800s. Potatoes were the
staple of the Irish diet, so when the fungus Phytophthora infestans infected
potatoes, causing them to rot in the fields, the result was devastating. By 1846, the
potato harvest was so meager that starvation and hunger-based diseases were
widespread. An estimated 1,240,000 people had died, and 1,200,000 more had
migrated to other countries. Potato blight remains an economic threat to potato
farmers even today.

1.2 Disease and Warfare

Warfare and infectious diseases have always been intimately connected. The poor
sanitation, movement of peoples, and malnutrition that a war brings, all foster
outbreaks of disease. For example, in 1812 when Napoleon invaded Russia he lost
more of his troops to typhus (a bacterial disease) than to all other causes combined.
And more of his soldiers died from wound-related bacterial infections (such as
tetanus and gas gangrene) than from the wound itself.

1.3 Optical Visual Aids

The first vision aid was invented (inventor unknown- possibly a monk) called a
reading stone around 1000 AD. It was a glass sphere that magnified when laid on
top of reading materials. In the year 1590, two Dutch eye-glass makers, Zaccharias
Janssen and son Hans Janssen experimented with multiple lenses placed in a tube.
The Janssens observed that viewed objects in front of the tube appeared greatly
enlarged, creating both the forerunner of the compound microscope and the
telescope.

1.3.1 Microscopy

Robert Hooke (1635-1703): Robert Hooke, a young English scientist, became the
first person to view and describe fungi using a simple compound microscope. In
1665, Hooke published Micrographia, which detailed his observations of tiny cork-
like cells resembling ‘little boxes’. Hooke is known principally for his law of
elasticity (Hooke's Law) and for his work as ‘the father of microscopy’ - it was
Hooke who coined the term ‘cell’ to describe the basic unit of life.
Antony van Leeuwenhoek (1632-1723): Antony van Leeuwenhoek, a Dutch
merchant, made small hand-held microscopes as a hobby. Squinting through the
lens at specimens held on a pin, he discovered a world of invisible creatures he
called animalcules (small animals). He found them almost everywhere he looked:
in water droplets, particles of soil and his teeth scrapings. In 1674, Leeuwenhoek
communicated his discoveries to the Royal Society of London, sending detailed
drawings. His primary observations shook the scientific community and led to
expanded use of microscopy as a standard scientific tool (Fig. 1.1). All his
drawings and nine of the estimated 500 microscopes that Leeuwenhoek made still
exist. The most powerful of these has a magnification of 266, powerful enough to
magnify an average-sized bacterial cell to the size of the period at the end of this
sentence.
 Fig. 1.1 Leeuwenhoek’s microscope

1.4 Spontaneous Generation

The idea that life routinely arises from non-life was supported by Aristotle (Circa
350 BC). According to him, 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. From ancient times,
spontaneous generation was thought to be the origin of many organisms (such as
rats and flies) that routinely appeared in certain materials. Microorganisms always
appeared suddenly in certain materials (meat juices or plant extracts, for example)
that had previously been free of them. It seemed logical that microbes were
products of spontaneous generation (the formation of living things from inanimate
matter).
Needham versus Spallanzani: For 80 years the above debate continued. Then the
proponents of spontaneous generation seemed to gain ground when in 1745 an
English clergyman named John Needham did a well-publicized experiment.
Everyone knew boiling killed microorganisms. Therefore, he boiled chicken broth,
put it in a flask, and sealed it. Microorganisms could develop in it only by
spontaneous generation. Experiments with gravy seemed to show that life could be
generated from non-living materials.
But an Italian priest and professor named Lazzaro Spallanzani was not convinced.
According to him, perhaps microorganisms entered the broth after boiling but
before sealing. Therefore, Spallanzani put broth in a flask, sealed it, and then
boiled it. No microorganisms appeared in the cooled broth. Still the critics were not
persuaded. Spallanzani didn’t disprove spontaneous generation, they said, he just
proved that spontaneous generation required air.
Gradually, spontaneous generation was rejected as the origin of visible organisms.
Francesco Redi, a physician in Italy, played a major role. In 1665 he did
experiments with covered and uncovered jars of meat. He showed that maggots
(fly larvae) developed only in meat that flies could reach to lay eggs on.
Apparently, spontaneous generation did not occur, at least in the case of flies.
Instead, flies and by extension all living things come only from preexisting living
things. Still, many people believed that microorganisms were an exception to this
rule. They are very simple and they always appear, in large numbers, soon after a
plant or animal dies. Could decomposition form microorganisms instead of
microorganisms causing decomposition? Controversy of spontaneous generation
gained momentum during the late 18th and 19th centuries, when further advances
in microscopy allowed people to view bacteria and other microorganisms.
1.5 Pasteur’s Experiments

Remarkably, the controversy continued another 100 years and became a significant
barrier to the development of microbiology as a science. Finally, in 1859 the
French Academy of Science sponsored a competition to prove or disprove the
theory of spontaneous generation of microbes. A young French chemist named
Louis Pasteur (1822-1895) entered to counter the argument that air was necessary
for spontaneous generation. Pasteur used barriers that allowed free passage of air
but not microorganisms. In his most famous experiment, Pasteur boiled meat broth
in a flask and then drew out and curved the neck of the flask in a flame in the shape
of a swan’s neck (Fig. 1.2). No microorganisms grew in the flask. But when he
tilted the flask so some broth flowed into the curved neck and then tilted it back so
the broth was returned to the base of the flask, the broth quickly became cloudy
with the growth of microbial cells. Gravity had caused the microbial cells that had
entered the flask to settle at the low point of the neck. They never reached the broth
in the base until they were washed into it. Thus, Pasteur convinced the scientific
world that spontaneous generation of microorganisms does not occur even in the
presence of air. Pasteur’s simple but elegant experiments grounded microbiology
in scientific reality. Microorganisms could now be studied by rational scientific
means. Probably the most famous contribution to microbiology by Pasteur is the
heating process he developed to kill spoilage microbes while still preserving
flavor.

Fig. 1.2 Pasteur’s swan-neck flask

1.6 Fermentation

Another important contribution of Pasteur was defining fermentation. In 1856, the

father of one of Pasteur's chemistry students asked him to help him solve some
problems he was encountering in his attempt to make alcohol by fermenting
beetroot. Often, instead of alcohol, the fermentations yielded lactic acid. At the
time, fermentation was believed to be a pure chemical process in which sugar was
transformed into alcohol. But in 1857, Pasteur proved that a microscopic plant
caused the souring of milk (lactic acid fermentation). Pasteur was able to prove
that living cells, the yeast, were responsible for forming alcohol from sugar, and
that contaminating microorganisms found in ordinary air could turn the ferments
sour. Next, he identified the microorganisms responsible for both normal and
abnormal fermentations, and found that through heating wine, beer, milk, or
vinegar briefly, certain living organisms could be killed, thereby sterilizing or
'pasteurizing' the substances. He reported his findings in "Mémoire sur la
fermentation appelée lactique" (Memoir on the fermentation of lactic acid) in
1857, and "Mémoire sur la fermentation alcoholique" (Memoir on the
fermentation of alcohol) in 1860. Pasteur also made notable contributions in the
field of vaccination and immunity. Studying cholera, Pasteur found that attenuated
organisms, inoculated into poultry, offered protection against virulent strains.
Based on this research, he developed the first rabies and anthrax vaccines.

1.7 Immunology

Edward Anthony Jenner (1749-1823) was an English scientist who studied his
natural surroundings in Berkeley, Gloucestershire. Jenner is widely credited as the
pioneer of smallpox vaccine, and is sometimes referred to as the 'Father of
Immunology'. Though Pasteur's achievements in microbial immunity were
revolutionary, Jenner is credited with inventing the first vaccine against smallpox
in late 1700s. In 1796, Jenner developed a controversial experiment to determine
the validity of rumours that were circulating in rural communities. Milkmaids and
villagers often recanted, "if you want to marry a woman who will never be scarred
by the pox, marry a milkmaid." Jenner speculated that becoming infected with
cowpox could offer protection against the more virulent smallpox. To test his
hypothesis, he created an inoculation with scrapings of cowpox lesions from the
fingers of Sarah Nelmes, a young milkmaid and injected it into an 8-year old boy
named James Phipps. As expected, James developed the mild fever and cowpox
lesions typical of the disease. After a few weeks of recovery, Jenner injected James
with the live smallpox virus and found that the boy was indeed protected from the
disease. In 1798, Jenner published his findings and presented them to the Royal
Society.

1.8 Germ Theory of Disease

Once spontaneous generation of microbes was disproved, microbiology exploded.

It changed from an observational science to an experimental science. The way was
opened to study the cause of infectious diseases. Building on Pasteur’s work, a
German physician, Robert Koch, proved that microorganisms (germs, as they were
and are still sometimes called) cause disease. He showed further that specific
microorganisms cause specific diseases. Koch also introduced higher scientific
standards of rigor to microbiology, as exemplified by those called Koch’s
postulates.

1.8.1 Koch’s postulates

In 1876 Robert Koch while studying anthrax, a disease of cattle and sheep that also
affects humans established “scientific rules” to show a cause and effect
relationship between a microbe and a disease known as Koch’r postulates as
follows:
 The same organisms must be found in all cases of a given disease.
 The organism must be isolated and grown in pure culture.
 The isolated organism must reproduce the same disease when inoculated
into a healthy susceptible animal.
 The original organism must again be isolated from the experimentally
infected animal.
During his work on anthrax, Koch made another critically important contribution
to microbiology. He developed a technique to obtain a pure culture of a bacterium
(one that contains only a single kind of bacteria) and propagate it. In nature many
kinds of bacteria are found growing together (mixed cultures). It is difficult to do
valid experiments using mixed cultures because they are so complicated, although
such studies are now becoming more common.

1.9 Antiseptic Surgery

Joseph Lister (1827-1912), a British surgeon was the pioneer of antiseptic surgery,
who promoted the idea of sterile surgery while working at the Glasgow Royal
Infirmary. Lister successfully introduced carbolic acid to sterilise surgical
instruments and to clean wounds, which led to reduced post-operative infections
and made surgery safer for patients.

1.10 Sterilization

In 1877, John Tyndall published his method for fractional sterilization and clarified
the role of heat resistant factors (spores) in putrefaction. Tyndall's conclusion
added a final footnote to the work of Pasteur and others in proving that
spontaneous generation is impossible. For some of his experiments on light and
gases he needed to cleanse the air of particles and so developed a completely novel
way to assay the purity of air. ‘The eye being kept sensitive by darkness’, Tyndall
reported, ‘a concentrated beam of light was found to be a most searching test for
suspended matter – a test indeed indefinitely more searching than that furnished by
the most powerful microscope’. Using his new piece of equipment, which relied on
observing light scattered by dust particles, Tyndall quickly made a series of
important observations on the properties of the ‘floating matter’ of air.

1.11 Milk Fermentation

In 1878, Joseph Lister published his study of lactic fermentation of milk,

demonstrating the specific cause of milk souring. His research was conducted
using the first method developed for isolating a pure culture of a bacterium, which
he named Bacterium lactis.

1.12 Virology

Virology, the study of viruses, began in 1892, when the Russian microbiologist
Dmitri Iwanowski discovered the tobacco mosaic virus. Iwanowski was studying a
disease of tobacco plants called tobacco mosaic disease. To identify its cause, he
forced juice from diseased plants through filters that retained the smallest bacteria.
He found the filtered juice still caused disease. Because bacteria were believed to
be the smallest microorganisms, Iwanowski first thought his methodology might be
flawed. But repeated experimentation convinced him that minute disease-causing
agents were passing through the filter. He called these tiny agents ‘filterable
viruses’. They could not be seen, even under the most powerful microscopes of
that time. Until the electron microscope was developed in the 1930s, we knew
viruses existed.

1.12.1 Bacteriophage

In 1915, the first discovery of bacteriophage was done by Frederick Twort. Twort's
discovery was something of an accident. He had spent several years growing
viruses and noticed that the bacteria infecting his plates became transparent. Later
on in 1917 Felix d'Herrelle independently described bacterial viruses and coined
the name ‘bacteriophage’. In 1926, Thomas Rivers distinguished between bacteria
and viruses, establishing virology as a separate area of study.