Section A: Basic Microbiology

1
SCOPE AND HISTORICAL DEVELOPMENTS
IN MICROBIOLOGY
“Science contributes to our culture in many ways, as a
creative intellectual activity in its own right, as a light which
has served to illuminate man’s place in the uni-verse, and
as the source of understanding of man’s own nature”
—John F. Kennedy (1917–63)
The President of America

The bacterium Escherichia coli

INTRODUCTION AND SCOPE

MICROBIOLOGY is a specialized area of biology (Gr. bios-life+ logos-to study) that concerns with the
study of microbes ordinarily too small to be seen without magnification. Microorganisms are
microscopic (Gr. mikros-small+ scopein-to see) and independently living cells that, like humans, live
in communities. Microorganisms include a large and diverse group of microscopic organisms that exist
as single cell or cell clusters (e.g., bacteria, archaea, fungi, algae, protozoa and helminths) and the
viruses, which are microscopic but not cellular. While bacteria and archaea are classed as prokaryotes
(Gr. pro-before+ karyon-nucleus) the fungi, algae, protozoa and helminths are eukaryotes (Gr. eu-true
or good+ karyon-nucleus). Microorganisms are present everywhere on earth, which includes humans,
animals, plants and other living creatures, soil,water and atmosphere.
Microorganisms are relevant to all of our lives in a multitude of ways. Sometimes, the influence
of microorganisms on human life is beneficial, whereas at other times, it is detrimental. For
example, microorganisms are required for the production of bread, cheese, yogurt, alcohol, wine,
beer, antibiotics (e.g., penicillin, streptomycin, chloramphenicol), vaccines, vitamins, enzymes and
many more important products as shown in the Tables 1.1, 1.2, and 1.3. Many products of microbes
contribute to public health as aids to nutrition, other products are used to interrupt the spread of
disease, still others hold promise for improving the quality of life in the year’s ahead.

1
2 A TEXTBOOK OF BASIC AND APPLIED MICROBIOLOGY

Table 1.1: Major antibiotics and their microbial sources

Antibiotic Microbial source

Bacitracin Bacillus licheniformis

Cephalosporin Cephalosporium acremonium
Chloramphenicol Streptomyces venezuelae
Cycloheximide Streptomyces griseus
Cycloserine Streptomyces orchidaceus
Erythromycin Streptomyces erythraeus
Griseofulvin Penicillium griseofulvum
Kanamycin Streptomyces kanamyceticus
Lincomycin Streptomyces lincolnensis
Neomycin Streptomyces fradiae
Nystatin Streptomyces noursei
Penicillin Penicillium chrysogenum
Polymyxin B Bacillus polymyxa
Streptomycin Streptomyces griseus
Teicoplanin Actinoplanes teichomyceticus
Tetracycline Streptomyces rimosus
Vancomycin Streptomyces orientalis

Table 1.2: Major industrial enzymes from bacteria, molds and yeasts and their applications

Enzyme Microorganism Application

Bacterial Enzymes
Amylase (α and β ) Bacillus Starch coatings (paper), desizing
(textiles), removal of stains, detergents
(drycleaning)
Glucose isomerase Bacillus, Streptomyces Fructose syrup
Penicillin amidase Bacillus Pharmaceutical
Protease Bacillus Detergent, spot removing, desizing,
wound cleaning

Mold Enzymes
α-Amylase Aspergillus Baking (Bread)
Glucoamylase Aspergillus, Rhizopus Syrup and glucose manufacture,
digestive aid (pharmaceutical)
Rennet (aspartic proteinases) Mucor miehei Cheese
Pectinase Aspergillus, Sclerotinia Drinks
Protease (aspartic proteinases) Aspergillus Baking
Cellulase Aspergillus, Trichoderma Liquid, coffee concentrates, digestive
aid, degradation of wood or wood by-
products
 SCOPE AND HISTORICAL DEVELOPMENTS IN MICROBIOLOGY 3

Enzyme Microorganism Application

α-Galactosidase Aspergillus niger Pharmaceutical (helps in digestion

(commercial name Beano) of sugar in humans)
Yeast Enzymes
Invertase Saccharomyces Confectionary
Lactase (β-galactosidase) Kluyveromyces Dairy
Raffinase (α-galactosidase) Saccharomyces Food

Table 1.3: Fermented foods from microorganisms

Fermented Substrate Microorganism Country/

Food region

Idli Rice and Urad bean Leuconostoc mesenteroides, India

Streptococcus faecalis
Ang-kak Rice Monascus purpureus China
Gari Cassava Corynebacterium West Africa
manihot, Geotrichum
candidum
Kaffir beer Sorghum caffrorum Lactobacillus delbrueckii South Africa
or Eleusine coracana Saccharomyces cerevisiae
Kefir Milk Lactobacillus and Yeast Russia
Yoghurt Milk Streptococcus ther- Worldwide
mophilus, Lactobacillus
bulgaricus
Cheese Milk Penicillium roqueforti Worldwide
P. camemberti

Microbes are also an important and essential component of an ecosystem. Molds and bacteria
play key roles in the cycling of important nutrients in plant nutrition particularly those of
carbon, nitrogen and sulphur. Bacteria referred to as nitrogen fixers live in the soil where they
convert vast quantities of nitrogen in air into a form that plants can use. Microorganisms also
play major roles in energy production. Natural gas (methane) is a product of bacterial activity,
arising from the metabolism of methanogenic bacteria. Microoragnisms are also being used to
clean up pollution caused by human activities, a process called bioremediation (the introduction
of microbes to restore stability to disturbed or polluted environments). Bacteria and fungi
have been used to consume spilled oil, solvents, pesticides and other environmentally toxic
substances.
Microorganisms have also harmed humans and disrupted societies over the millennia. Microbial
diseases undoubtedly played a major role in historical events, it was in the year 1347 when plague
or ‘black death’ struck Europe and within 4 years killed 25 million people, that is, one third of
the population. Some of the common human diseases caused by bacteria, fungi (molds and yeasts),
protozoa, helminths are shown in the Tables 1.4–1.7.
4 A TEXTBOOK OF BASIC AND APPLIED MICROBIOLOGY

Table 1.4: Human diseases caused by bacteria

Type Species Disease

Spirochetes Treponema pallidum Syphilis

Borrelia recurrentis Relapsing fever
Borrelia burgdorferi Lyme disease
Leptospira interrogans Leptospirosis

Helical, vibrioid, Gram- Campylobacter jejuni Food borne campylobacter enteritis

negative bacteria Helicobacter pylori Peptic ulcer and chronic gastritis
(=Campylobacter pylori)

Gram-negative aerobic Legionella pneumophila Legionnaires’ disease

rods and cocci Neisseria gonorrhoeae Gonorrhoea
Neisseria meningitidis Meningococcal meningitis
Brucella melitensis Brucellosis
Bordetella pertussis Whooping cough
Francisella tularensis Tularemia (Rabbit fever)

Facultatively aerobic, Escherichia coli Oppurtunistic infections

Gram-negative rods Salmonella typhi
Shigella dysenteriae
Klebsiella pneumoniae
Proteus sp.
Yersinia pestis
Vibrio cholerae
Haemophilus influenzae
Gardnerella vaginalis
Typhoid fever
Bacillary dysentry (Shigellosis)
Pneumonia, Meningitis
Urinary tract infections
Bubonic plague
Cholera
Meningitis, Ear infections
Vaginitis

Rickettsias and Rickettsia rickettsiae Rocky mountain spotted fever

Chlamydias Rickettsia prowazekii Epidemic typhus
Rickettsia typhi Murine typhus
Coxiella burnetii Q-Fever
Chlamydia trachomatis Trachoma
Chlamydia psittaci Ormithosis (Psittacosis)
Chlamydia pneumoniae Pneumonia

Mycoplasmas Mycoplasma pneumoniae Primary atypical pneumonia

Gram-positive cocci Staphylococcus aureus Boils, wound infections, Toxic shock

Streptococcus pneumoniae
Streptococcus pyogenes
Streptococcus mutans
syndrome, Food poisoning, Impetigo
Pneumococcal pneumonia
Strep throat, Glomerulonephritis,
Rheumatic fever, Impetigo
Dental caries

(Contd.)
 SCOPE AND HISTORICAL DEVELOPMENTS IN MICROBIOLOGY 5

Type Species Disease

Spore forming Gram- Bacillus anthracis Anthrax

positive rods and cocci Clostridium tetani Tetanus
Clostridium perfringens Gas gangrene
Clostridium botulinum Botulism
Clostridium difficile Pseudomembranous colitis

Regular, non-sporing Lactobacillus sp. Normal human flora

Gram-positive rods Listeria monocytogenes Listeriosis

Irregular, non-sporing, Corynebacterium diphtheriae Diphtheria

Gram-positive rods Propionibacterium acne Acne

Mycobacteria (Acid- fast Mycobacterium tuberculosis Tuberculosis

organisms) Mycobacterium leprae Leprosy

Table 1.5: Human diseases caused by fungi

Disease Pathogen

(A) Superficial mycoses

Black piedra Piedraia hortae
White piedra Trichosporon beigelii
Dandruff or Tinea versicolor Malassezia furfur (Pityrosporum ovale)

(B) Dermatomycoses ( cutaneous mycoses)

Tinea capitis (Ringworm) Microsporum audouinii
Tinea pedis (Athlete’s foot) Trichophyton spp.
Tinea cruris (Jock itch) Epidermophyton floccosum
Tinea unguium (Ringworm of nails) Trichophyton rubrum

(C) Subcutaneous mycoses

Chromoblastomycosis Fonsecaea pedrosoi (Phialophora verrucosa)
Maduromycosis Madurella mycetomatis
Sporotrichosis Sporothrix schenckii

(D) Systemic mycoses (deep mycoses)

Blastomycosis Blastomyces dermatitidis (Ajellomyces dermatitidis)
Coccidioidomycosis (valley fever) Coccidioides immitis
Cryptococcosis Cryptococcus neoformans (Filobasidiella neoformans)
Histoplasmosis Histoplasma capsulatum
(Contd.)
6 A TEXTBOOK OF BASIC AND APPLIED MICROBIOLOGY

Disease Pathogen

(E) Opportunistic mycoses

Aspergillosis Aspergillus fumigatus
Candidiasis (oral, napkin (diaper) Candida albicans
candidiasis, Candidal vaginitis)
Pneumocystis pneumonia (PCP) Pneumocystis jiroveci (P.carinii)
Zygomycosis Mucor and Rhizopus spp.

(F) Food poisoning

Ergotism (ergot poisoning) Claviceps purpurea

Table 1.6: Human diseases caused by protozoans

Phylum Pathogen Disease

Apicomplexa Babesia microti Babesiosis

Plasmodium falciparum, Malaria
P. ovale, P. vivax, P. malariae
Toxoplasma gondii Toxoplasmosis
Cryptosporidium parvum Cryptosporidiosis

Rhizopoda Acanthamoeba sp. Amoebic keratitis

Entamoeba histolytica Amoebic dysentery
Naegleria fowleri Microencephalitis

Mastigophora (Flagellata) Giardia lamblia (G. Intestinalis) Giardiasis

Trichomonas vaginalis Protozoal vaginitis
Trypanosoma brucei African sleeping sickness
Trypanosoma cruzi Chaga’s disease

Ciliophora (Ciliata) Balantidium coli Balantidial dysentery

Table 1.7: Human diseases caused by helminths

Phylum Pathogen Disease

Platyhelminthes Paragonimus westermanni Paragonimiasis

(Lung fluke)
Schistosoma sp. (Blood flukes) Schistosomiasis
Clonorchis sinensis Clonorchiasis
(Chinese liver fluke)

(Contd.)
 SCOPE AND HISTORICAL DEVELOPMENTS IN MICROBIOLOGY 7

Phylum Pathogen Disease

Taenia saginata (Beef tapeworm) Taeniasis

Taenia solium (Pork tapeworm) Taeniasis
Hymenolepsis nana Hymenolepasis
(Dwarf tapeworm)
Diphyllobothrium latum Diphyllobothriasis
(Fish tapeworm)
Echinococcus granulosus Echinococcosis
(Dog tapeworm)
Fasciola hepatica Fascioliasis
(Sheep liver fluke)

Nematoda (Roundworms) Strongyloides stercoralis Strongyloidiasis

(Threadworm)
Ascaris lumbricoides Ascariasis
(roundworm)
Necator americanus (hookworm) New world hookworm disease
Ancylostoma duodenale Old world hookworm disease
(hookworm)
Enterobius vermicularis Pinworm feotalism
(Pinworm)
Trichuris trichiura Trichuriasis
(Whipworm)
Trichinella spiralis Trichinosis
(Trichinaworm)
Wuchereria bancrofti Elephantiasis or bancroftian
filariasis
Dirofilaria immitis (Heartworm) Filariasis

HISTORICAL DEVELOPMENTS IN MICROBIOLOGY

The beginnings
The study of microorganisms, or microbiology began when the first microscopes were developed in
1665 by the English scientist, Robert Hooke who viewed many small objects and structures using a
simple lens that magnified approximately 30 times. His specimens included the eye of a fly, a bee
stinger, and the shell of a protozoan. Hooke also examined thin slices of cork, which was the bark of
a particular type of oak tree. He found that cork was made of tiny boxes that Hooke referred to as
‘cells’. He published his work in a book Micrographie which contained a miscellany of his thoughts
on chemistry as well as a description of the microscope and its uses. Hooke in 1665 described the
fruiting structures of molds. Thus, Robert Hooke was the first person to describe microorganisms.
 8 A TEXTBOOK OF BASIC AND APPLIED MICROBIOLOGY

MICROFOCUS 1.1
Antony van Leeuwenhoek (pronounced Layu-wen- hoek) was born on
October 24, 1632 in Delft, Holland (now Netherlands). In 1674, he made first
observation of microoraganisms and was the first person to observe and
accurately describe and measure bacteria and protozoa, termed by him, as
“animalcules” which he thought were tiny animals. In 1677, he became the
first person to describe spermatozoa and was one of the earliest to describe
red blood corpuscles. In 1680, he was elected a fellow of the Royal Society
of London, and with Isaac Newton and Robert Boyle, he became one of the
first famous men of his time. He died on August 30, 1723 at the age of 90.
Because of his extraordinary contribution to microbiology, he is considered
as the father of bacteriology and protozoology. Antony van
Leeuwenhoek
(1632-1723)

Unicellular life was first described just a few years after Hooke recorded his observations of
the microscopic world. Antony van Leeuwenhoek (Microfocus 1.1) was a Dutch merchant who
polished grains of sand into lenses which were able to magnify 300 times and added a simple focus
mechanism. With his microscope, van Leeuwenhoek viewed rain and pond water, infusions made
from peppercorns, and scrapings from his teeth in the year 1674 and termed the tiny microorganisms
as ‘animalcules’. In 1676, van Leeuwenhoek sent his drawings to the Royal Society of London. This
has special significance to microbiology because it contained his first detailed description of the
microorganism.

The transition period

Biology of the 1700s was a body of knowledge without a focus. It consisted of observations of plant
and animal life and the attempts by scientists to place the organisms in logical order. The dominant
figure of the era was Carolus Linnaeus (1707–1778), a Swedish botanist who brought all the plant
and animal forms together under one Binomial nomenclature (naming of an organism by two
names—the genus and species) system of classification scheme. His book, Systema naturae, was
first published in 1735.
Discovery of the microscopic world raised some interesting queries and eventually led
scientists to question some of the long-held beliefs. At that time in history, the scientific community
used a theory known as ‘spontaneous generation’ (the doctrine that holds that lifeless objects
give rise to living organisms) to explain the apparently magical origins of life. The theory proposed
that simple life forms arose spontaneously from non-living materials and had its basis in the
findings of Aristotle in the fourth century BC.
Although most people accepted spontaneous generation, the theory did have some strong
opponents. Among the first to dispute the theory of spontaneous generation was the Italian
scientist, Francesco Redi (1626–1697). He reasoned that flies had reproductive organs while
observing van Leeuwenhoek’s drawings. He suggested that flies land on pieces of exposed meat
and lay their eggs, which then hatch to maggots. This would explain the ‘spontaneous’ appearance
of maggots. In the 1670s, Redi performed a series of tests in which he covered jars of meat with
fine lace, thereby preventing the entry of flies. The meat would not produce maggots as it was
protected and Redi temporarily put to rest the notion of spontaneous generation.
 SCOPE AND HISTORICAL DEVELOPMENTS IN MICROBIOLOGY 9

Although Redi’s work became widely known, the doctrine of spontaneous generation was too
firmly entrenched to be abandoned. In 1748, British clergyman, John Needham (1713–81) put forth
the notion that in flasks of mutton gravy, microorganisms arise by spontaneous generation. He even
boiled several flasks of gravy and sealed the flasks with corks as Redi had sealed his jars. Still,
the microorganisms appeared.
Italian scientist Abbe Lazzaro Spallanzani (1729–99) criticized Needham’s work. In 1767,
Spallanzani boiled meat and vegetable broths for long period of time and then sealed the necks
by melting the glass. As control experiments, he left some flasks open to the air, stoppered some
loosely with corks, and boiled some briefly, as Needham had done. After two days, he found the
control flasks swarming with organisms, but the sealed flasks had no organisms. Needham
countered that Spallanzani had destroyed the ”vital force” of life with excessive amounts of heat.
While the spontaneous generation was being debated, some of the scientists were concerned
about the transmission of the disease. In 1546, Italian scientist Girolamo Fracastoro held the concept
that “contagion is an infection that passes from one thing to another”. He recognized three forms of
passage, namely contact, lifeless objects, and air (Table 1.8). This notion received little credibility that
microorganisms were the substance of contagion. The German Athanasius Kircher was paid little
attention when he reported “microscopic worms” in the 1600s in the blood of plague victims.
Christian Fabricius was also neglected when he suggested in 1700s that fungi might be the cause of
rust and smut diseases in plants. Edward Jenner (Microfocus 1.2) was accorded honours in 1798
when he discovered immunization for smallpox, despite the fact that he could not explain the cause
of the disease. In 1847, Hungarian physician, Ignaz Semmelweis reported that blood poisoning
agent was transmitted to maternity patients by physicians fresh from performing autopsies in the
mortuary. Semmelweis showed that hand washing in chlorine water could stop the spread of disease.
His call for disinfection practices were however largely unheeded because it implied that physicians
were at fault.

MICROFOCUS 1.2

Edward Jenner, born in 1749, was an English physician from Berkeley,

Gloucestershire, England. His great gift to mankind was his vaccine for smallpox
(characterized by production of skin lesions called pox (pocks), caused by Variola,
belonging to the category of pox viruses). Jenner’s discovery, that a less
pathogenic agent could confer protection against a more pathogenic one, is
especially remarkable in view of the fact that microscopy was still in its infancy
and the nature of the virus was not known. The modern era of vaccines and
vaccination, thus began in 1798 with Edward Jenner’s use of cowpox as a
vaccine against smallpox.

Edward Jenner
(1749–1823)

John Snow, a British physician, traced the source of cholera to the municipal water supply of
London during an 1854 outbreak. He reasoned that by avoiding the contaminated water source,
people could avoid the disease. Snow’s recommendations were adopted and the spread of disease
was halted. Both Semmelweis and Snow drew attention to the fact that a poison or unseen object
in the environment was responsible for the disease, but the proof was still lacking. Joseph Lister
(Microfocus 1.3) in 1867, developed a system of antiseptic surgery designed to prevent micro-
organisms from entering wounds.
10 A TEXTBOOK OF BASIC AND APPLIED MICROBIOLOGY

MICROFOCUS 1.3

Joseph Lister was born in 1827. He developed a system of antiseptic surgery

designed to prevent microorganisms from entering wounds in 1867. In 1878,
Lister studied the lactic acid fermentation of milk and demonstrated the specific
cause of milk souring. He also developed a method for isolating a pure culture of a
bacterium, named as Bacterium lactis. Because of his notable contribution-first
introduction of principles of sterile surgery in medical practice, which was so far
reaching in its effects—Lister will always be known as the Father of antiseptic
surgery. He died at the age of 85 in the year 1912.
Joseph Lister
(1827–1912)

Table 1.8: Some early observations in microbiology before the dawn of golden era

Time period Investigator Observations

Fourth Century Aristotle Living things do not need parents, spontaneous generation
BC. apparently occurs.
Mid 1500s Fracastoro “Contagion” passes among individuals, objects and air.
Mid 1600s Kircher “Microscopic worms” are present in blood of plague victims.
Mid 1600s Francisco Redi Fly larvae arise by spontaneous generation.
Late 1600s Van Leeuwenhoek Microscopic organisms are present in numerous environ-
ments.
Early 1700s Christian Fabricius Fungi cause rust and smut diseases in plants.
Early 1700s Joblot Existence of various forms of protozoa.
Mid 1700s John Needham Microorganisms in broth arise by spontaneous generation.
Mid 1700s Lazzaro Spallanzani Heat destroys microorganisms in broth.
Late 1700s Edward Jenner Recoverers from cowpox do not contract smallpox.
Mid 1800s Ignaz Semmelweis Chlorine disinfection prevents disease spread.
Mid 1800s John Snow Water is involved in disease transmission.

The classical golden age of microbiology (1854–1914)

The science of microbiology blossomed during a period of about 60 years referred to as the Golden
Era of Microbiology. The period began in 1857 with the work of Louis Pasteur and continued into the
twentieth century until the advent of World War I. During this period, numerous branches of
microbiology were laid for the maturing process that has led to modern microbiology.
Louis Pasteur (Microfocus 1.4) was the first to report the role of microorganisms in fermentation
in 1848, he achieved distinction in organic chemistry for his discovery that tartaric acid, a four-
carbon organic compound, forms two different types of crystals. Pasteur successfully separated the
crystals while looking through the microscope. In 1854, at the age of 32, he was appointed Professor
of Chemistry at the University of Lille in northern France.
Pasteur in 1857 unravelled the mystery of sour wines. In a classic series of experiments, Pasteur
clarified the role of yeasts in fermentation of fruits and grains resulting in the production of alcohol.
 SCOPE AND HISTORICAL DEVELOPMENTS IN MICROBIOLOGY 11

He also found that bacteria were responsible for spoilage of wine. He firmly disproved the
spontaneous generation doctrine by his Swan-Neck Flask experiment (Fig. 1.1). He proposed germ
theory of disease and discovered the existence of life in the absence of free oxygen (anaerobic
growth). He showed that mild heating could be used to kill microorganisms in broth (pasteurization).
Pasteur suggested methods to control pebrine disease in silkworm, isolated the causative agent of
cholera (Vibrio cholerae) and rabies (Lyssa) virus and also developed anti rabies and anthrax
(Bacillus anthracis) vaccines.
Although Pasteur failed to relate a specific organism to a specific disease, his work stimulated
others to investigate the nature of microorganisms and to ponder their association with disease.
German botanist, Ferdinand Cohn (1828–98), discovered that bacteria multiply by dividing into
two cells. He also observed that certain bacteria form an extremely resistant structure called
endospore in the cell.

Fig. 1.1: Pasteur’s experiment with the swan-necked flasks to disprove spontaneous generation. (a) Life appeared
in broth in flasks exposed to air. (b) No life appeared in sealed flasks. (c) No life appeared in flasks where the neck
was continuously heated. (d) No life appeared in flasks when the microorganisms were trapped in the bend of the side
arm.
12 A TEXTBOOK OF BASIC AND APPLIED MICROBIOLOGY

MICROFOCUS 1.4

Louis Pasteur- Notable Contributions

1857 – Lactic acid fermentation is due to a microorganism
1860 – Yeasts are involved in alcoholic fermentation
1861 – Disproved the theory of spontaneous generation
1861 – Introduction of the terms aerobic and anaerobic for yeasts.
Production of more alcohol in the absence of oxygen during
sugar fermentation- The Pasteur Effect
1862 – Proposed germ theory of disease
1867 – Pasteur devised the process of destroying bacteria known
as pasteurization.
1881 – Development of anthrax vaccine. Resolved Pebrine
problem of silkworms.
Louis Pasteur
1885 – Development of a special vaccine for rabies (the Pasteur
(1822–1895) treatment)
Louis Pasteur, a French microbiologist, was born on December 27,
1822 in Dole, France. He studied at the French school, the Ecole Normale Superieure. In 1848, he achieved
distinction in organic chemistry for his discovery that tartaric acid, a four carbon organic compound
forms two different types of crystals. Using a microscope, Pasteur successfully separated the crystals and
developed a skill that would aid his later studies of microorganisms. In 1854, at the young age of 32, he
was appointed Professor of Chemistry at the University of Lille in northern France. He died in 1895, at
the age of 73.

Cohn described the entire life cycle of Bacillus (vegetative cell → endospore → vegetative cell). He
is credited with the use of cotton plugs for closing flasks and tubes to prevent the contamination of
sterile culture media. In 1866, Cohn studied the filamentous sulphur-oxidizing bacterium Beggiatoa
mirabilis and was the first to identify the small granules present in the cell that are of sulphur,
produced from the oxidation of H2S.
The definite proof of the germ theory of disease was offered by Robert Koch (Microfocus 1.5)
from East Russia, now part of Germany. Koch’s primary interest was anthrax, a deadly blood
disease in cattle and sheep. In 1875, he injected mice with the blood of diseased sheep and cattle.
He then performed meticulous autopsies and noted that the same symptoms appeared regularly.
He isolated a few rod shaped bacilli from a mouse’s blood by placing the bacilli in the sterile
aqueous humor from an ox’s eye. The symptoms of anthrax appeared within hours. Koch autopsied
the animals and found their blood swarming with bacilli. He reisolated the bacilli in sterile aqueous
humor. Koch’s procedures came to be known as Koch’s postulates (Fig. 1.2). The four postulates
are:
• The suspected microorganism must always be found in diseased but never in healthy
individuals.
• The microorganism must be isolated in pure culture (one free of all other types of microbes) on
a nutrient medium.
• The same disease must result when the isolated microorganism is inoculated into a healthy
host.
• The same organism must be reisolated from the experimentally infected host.
 SCOPE AND HISTORICAL DEVELOPMENTS IN MICROBIOLOGY 13

Fig. 1.2: The diagrammatic representation of the Koch’s criteria for proving that a specific microorganism causes a
specific disease, i.e., the Koch’s postulates.

MICROFOCUS 1.5

Notable contributions of Robert Koch

1876 – Koch demonstrated that anthrax is caused by Bacillus anthracis.
1877 – Methods for staining bacteria, photographing and preparing
permanent visual records on slides.
1881 – Koch developed solid culture media and the methods for studying
bacteria in pure cultures.
1882 – Isolated the bacterium—Mycobacterium tuberculosis—that causes
tuberculosis.
1882 – Use of agar as a support medium for solid culture in Koch’s lab by
Hesse.
1883 – Isolation of Vibrio cholerae, the cause of cholera.
1883 – Verification of the germ theory of disease by relating a specific Robert Koch
organism to the specific disease. (1843–1910)
1884 – Koch put forth his postulates—known as Koch’s postulates.
Robert Koch was born in Hanover, Germany in 1843. For his contributions on tuberculosis, Robert Koch
was awarded the 1905 Nobel Prize for Physiology or Medicine. He died in the year 1910 at the age of 67.
14 A TEXTBOOK OF BASIC AND APPLIED MICROBIOLOGY

Koch chanced to observe in 1880 that a slice of potato contained small masses of bacteria,
which he termed colonies. Colonies contained millions of just one kind of bacteria. Koch con-
cluded that bacteria could grow and multiply on solid surfaces, and he added gelatin to his broth
to prepare a solid culture medium. He then inoculated bacteria to the surface and set the medium
aside to incubate. When colonies of the same bacterium grew together, a pure culture (an
accumulation of one type of microorganism formed by the growth of colonies of the organism)
formed. Koch could now inoculate laboratory animals with a pure culture of bacteria and be certain
that only one species of bacterium was involved. His work also proved that bacteria, not toxins
in the broth were the cause of the disease.
Gelatin was replaced with agar as a solidifying agent in the culture media as suggested by
Fannie Eilshemius Hesse, wife of Walter Hesse, an assistant in the Koch’s lab. Petri dish was also
invented about this time by Julius Petri, one of Koch’s assistants. In 1881, Koch demonstrated his
pure culture techniques in the International Medical Congress.
Koch’s proof of the germ theory was presented in 1876. Within two years, Pasteur had verified
the proof and gone a step further. He reported that bacteria were temperature-sensitive because
chickens did not acquire anthrax at their normal body temperature of 420C but did so when the
animals were cooled down to 370C. He also recovered anthrax spores from the soil and pointed out
that cattle were probably infected during grazing. This explained the periodic recurrence of the
disease.
One of Pasteur’s more remarkable discoveries was made in 1880 when a group of inoculated
chickens failed to develop chicken cholera. He had been working on ways to enfeeble bacteria using
heat, different growth media, passages among animals, and virtually anything he thought might
weaken them. Finally, he had developed two cultures whose ability to cause disease was reduced.
The trick was to suspend the bacteria in a mildly acidic medium and allow the culture to remain
undisturbed for a long period of time.When it was inoculated to chickens and later followed by
a dose of lethal cholera bacilli, the animals did not become sick. This principle is the basis for the
use of many vaccines for immunity. Pasteur applied the principle to anthrax in 1881 and found
he could protect sheep against the disease.
Koch, isolated the tubercle bacillus, the cause of tuberculosis. In 1884, Koch’s associate George
Gafky, cultivated the typhoid bacillus, and that same year another coworker, Friederich Loeffler,
isolated the diphtheria bacillus. In later years, Koch’s coworker, Emil von Behring, successfully
treated diphtheria by injecting antitoxin, a blood product (preparation of antibodies) obtained from
animals given injections of the toxin. For his work, von Behring was awarded the first Nobel Prize
in Physiology or Medicine.In 1885, Pasteur reached the zenith of his carrier when he successfully
immunized young Joseph Meister against the dreaded disease rabies. Although he never saw the
agent of rabies, Pasteur was able to cultivate it in the brains of animals and inject the boy with
bits of the tissue. The experiment was a triumph for Pasteur because it fulfilled his dream of
applying the principles of science to practical problems. A comparison of Pasteur and Koch’s
achievements is given in the Table 1.9.

Other pioneers of microbiology

Shibasaburo Kitasato of Japan studied with Koch and successfully cultivated the tetanus bacillus,
an organism that grows only in the absence of oxygen. One of the Pasteur’s associates was Elie
Metchnikoff (Microfocus 1.6), who in 1884, published an account of phagocytosis, a defensive
process in which the body’s white blood cells engulf and destroy microorganisms.
 SCOPE AND HISTORICAL DEVELOPMENTS IN MICROBIOLOGY 15

Table 1.9: A comparison of contributions of Louis Pasteur and Robert Koch

Characteristic Louis Pasteur Robert Koch

Country of origin France Germany (Prussia)

Preparatory Chemistry Medicine
education
Initial Milk souring, beer, Cause of anthrax
investigations wine fermentations
Accomplishments • Proposed germ theory of disease • Proved germ theory of disease
• Disproved theory of spontaneous • Developed cultivation methods for
generation bacteria
• Developed immunization • Isolated bacterium that causes
techniques tuberculosis
• Resolved pebrine problem of • Developed staining methods for bacteria
silkworms
• Developed rabies vaccine • Investigated cholera, malaria, sleeping
sickness
Associates Roux, Yersin, Metchnikoff Gaffky, Loeffler, von Behring, Richard
Pfeiffer
Nobel Prize No 1905 Nobel Prize in Physiology or
Medicine

MICROFOCUS 1.6

Elie Metchnikoff, one of the associates of Louis Pasteur, was a Russian zoologist
who lived in Paris and did his work at the Institute Pasteur, France. He was
born in Kharkor priovince of Ukraine (USSR) in 1845. By the 1860s he had
completed his formal studies in Embryology from various Universities of
Kharkor, Russia, Germany and Italy. Metchnikoff coined the term
“phagocytosis” which literally means” the eating of cells”. In 1884, he published
account of phagocytosis, a defensive process in which the body’s white blood
cells (WBCs) engulf and destroy microorganisms. Thus, he formulated the
basic theory on which the science of immunology is founded: that the body is
protected from infection by leukocytes that engulf bacteria and other invading
organism (cellular immunity). He became an administrator to the Institute
Pasteur in 1888 and eventually became its director. He was awarded the Nobel
Prize in 1908. Metchnikoff’s notable contribution was on the Bacillus bulgaricus Elie Metchnikoff
therapy and his underlying concept of health. Metchnikoff belived that (1845–1916)
streptococci and lactobacilli in yogurt assume residence in the intestine and
replace organisms that contribute to aging. Despite eating large quantities of yogurt, Metchnikoff died
an early death, in 1916, at age seventy-one.

A Pasteur Institute scientist, Charles Nicolle, proved that typhus fever was transmitted by lice.
Albert Calmette, also of the Institute, developed a harmless strain of the Tubercle bacillus used for
immunization. Jules Bordet, a Belgian bacteriologist isolated the bacillus of pertussis (whooping
cough) and developed the complement fixation test, a procedure once widely used in the diagnosis
of disease.
16 A TEXTBOOK OF BASIC AND APPLIED MICROBIOLOGY

Ronald Ross, an English physician working in the Far East in 1898 proved that mosquitoes were
the vital link in malaria transmission. The discovery earned him the 1902 Nobel Prize. Another
Englishman, David Bruce, isolated the cause of undulant fever. Bruce also showed that tsetse flies
transmit sleeping sickness. A third British subject, Almroth Wright, described opsonins, the chemical
substances that promote phagocytosis in the body.
In 1897, the Tokyo physician Masaki Ogata reported that rat fleas transmit bubonic plague.
This discovery solved a centuries old mystery of how plague spread. A year later, Kiyoshi Shiga
isolated the bacterium that causes bacterial dysentery, an important intestinal disease. The
organism was later named Shigella.
The American microbiologists, Daniel E. Salmon and Theobald Smith, were among the first
to use heat killed bacteria for immunizations. Salmon later studied swine plague and lent his name
to Salmonella, the cause of typhoid fever. Smith showed that Texas fever, a disease of cattle, was
transmitted by ticks. The University of Chicago pathologist Howard Taylor Rickkets located the
agent of Rocky Mountain spotted fever in the human bloodstream and demonstrated its
transmission via ticks. Another American, William Welch, isolated the gas gangrene bacillus at his
laboratory at John Hopkins University. Walter Reed led a contingent to Cuba and pinpointed
mosquitoes as the insects involved in yellow fever transmission.
In addition, Winogradsky and Beijerinck began examining the role of non-infectious micro-
organisms in the soil and reported that microorganisms play an important role in nitrogen, sulphur
and carbon cycling as well as process of nitrogen fixation by symbiotic or free living soil bacteria.
Iwanowsky and Beijerinck provided the first evidence for virus as infectious agent.
The advent of World War I in 1914 signaled a dramatic pause in microbiology research and
brought to an end the Golden Era of Microbiology.

The era of chemotherapy and microbial genetics

Paul Ehrlich in collaboration with Sakahiro Hata, discovered the drug, Salvarsan, an arsenobenzol
compound in 1910 for the treatment of syphilis caused by Treponema pallidum. Ehrlich laid important
foundation of the era of chemotherapy which is defined as the use of chemicals that selectively
inhibit or kill pathogens without causing damage to the victim.
Gerhard Domagk of Germany in 1935 reported that Prontosil, a red dye used for staining
leather, was active against pathogenic streptococci and staphylococci in mice even though it had
no effect against the same infectious agent in the test tube. The two French scientists Jacques and
Therese Trefonel in the same year showed that the compound Prontosil was broken down within
the body of the animal to sulphanilamide (sulpha drug) which was the true active factor. Domagk
was awarded Nobel Prize in 1939 for the discovery of the first sulpha drug.
The credit for the discovery of the first”wonder drug”, penicillin goes to a Scottish physician
and bacteriologist, Sir Alexander Fleming (Microfocus 1.7) in 1929 from the mold Penicillium
notatum. Fleming discovered the first antibiotic which is a microbial product that can kill
susceptible microorganisms and inhibit their growth. Sir Howard. W. Florey and Ernst B.
Chain at Oxford University in 1941 developed methods for industrial production of penicillin
in England. Fleming, Florey and Chain shared the Nobel Prize in 1945 for the discovery and
production of penicillin.
 SCOPE AND HISTORICAL DEVELOPMENTS IN MICROBIOLOGY 17

MICROFOCUS 1.7

Alexander Fleming, a Scottish, was born in the year 1881. He was a

physician by training, but spent most of his time studying bacteria. Sir
Alexander Fleming, in 1922 discovered that lysozyme, an enzyme
found in tears, saliva and sweat, could kill bacteria, the first body
secretion shown to have chemotherapeutic properties. He in 1928
discovered the first antibiotic (Gr. anti-against + bios- life, the microbial
product that can kill susceptible microorganisms or inhibit their
growth), penicillin. In 1929, Alexander Fleming published his findings
in the paper describing penicillin and its effect on Gram-positive
bacteria. Fleming died in 1955, at the age seventy-four.
Alexander Fleming
(1881–1955)

At the time of World War II (1939–44), S. A. Waksman of Rutgers’ University, USA discovered
another antibiotic, streptomycin along with Albert Schatz in 1944 from an actinomycete, Streptomyces
griseus. Waksman received the Nobel Prize in 1952 for his notable contribution and for the discovery
of streptomycin used in the treatment of tuberculosis, a bacterial disease caused by Mycobacterium
tuberculosis, that had been discovered by Robert Koch in 1882.
Dr. Paul R. Burkholder in 1947 isolated chloramphenicol (chloromycetin) from Streptomyces
venezuelae. Dr. B.M. Dugger in 1948 identified aureomycin from Streptomyces aureofaciens and
terramycin was discovered by Finlay, Hobby and collaborators in 1950 from Streptomyces rimosus.
Antibiotic production continues to be the important area of industrial research. Currently, there are
over 8000 antibiotics known, of which only a few are being used as chemotherapeutic agents.
In 1943, Italian microbiologist Salvador Luria and the German physicist Max Dulbriick carried
out a series of experiments with bacteria and viruses. They used the bacterium Escherichia coli to address
a basic question regarding the nature of mutations, spontaneous or induced. Luria and Dulbriick
showed that bacteria could develop spontaneous mutations that generate resistance to viral infection.
Besides the significance of their findings to microbial genetics, their use of E. coli as a microbial model
system showed to other researchers that these relatively simple microorganisms could be used to study
general principles of biology. The experiments carried out by Americans George Beadle and Edward
Tatum, using the fungus, Neurospora, showed that one gene codes for one enzyme i.e., one-gene one-
enzyme hypothesis. Oswald Avery, Colin Mcleod, and Maclyn McCarty, working with the bacterium
Streptococcus pneumoniae, suggested that deoxyribonucleic acid (DNA) is the genetic material in cells.
In 1953, American biochemist Alfred Hershey and geneticist Martha Chase, using bacterial viruses,
provided irrefutable evidence that DNA is the substance of genetic material. Joshua Lederberg
(Microfocus 1.8) in 1958 received the Nobel Prize in Physiology or Medicine for his discoveries
concerning genetic recombination and organization of genetic material in bacteria.
The small size of bacteria hindered scientists’ abilities to confirm that bacteria were “cellular” in
function. In the 1940s and 1950s, an electron microscope was developed that could magnify objects
and cells thousands of times more than typical light microscopes. With the electron microscopes, for
the first time bacteria were seen as being cellular like all other microbes, plants and animals. However
studies showed that they were organized in a fundamentally different way from other organisms. It
was shown that animal and plant cells contained a cell nucleus that stores the genetic information in
the form of chromosomes and was separated physically from other cell structures by a membrane
18 A TEXTBOOK OF BASIC AND APPLIED MICROBIOLOGY

envelope. This type of cellular organization is called eukaryotic (eu= true+karyon = kernel, nucleus).
Microscopic observations of the Protista and Fungi had revealed that these organisms also had a
eukaryotic organization.

MICROFOCUS 1.8
Dr. Joshua Lederberg was born on May 23, 1925 in Montclair, New Jersey.
Joshua Lederberg is noted for two landmark discoveries in bacterial genetics:
bacterial conjugation and transduction, both laying foundations for genetic
engineering, modern biotechnology and genetic approaches to medicine.
Interdisciplinary in his scientific interests and methods, he became a pioneer
of Exobiology and the exploration of space, and was instrumental in
introducing computers and artificial intelligence into laboratory research and
biomedical communication. Lederberg, along with Beadle and Tatum, was
awarded the Nobel Prize at the age 33, for his discoveries concerning genetic
recombination and organization of the genetic material of bacteria. In addition
to receiving the Nobel Prize, Lederberg has received many other awards and
honours. It can only be said that Joshua Lederberg single-handedly changed Dr. Joshua Lederberg
the nature of bacterial genetics and changed the course of both genetics and
biochemistry.

Studies with the electron microscope revealed that bacterial cells had few of the cellular structures
typical of eukaryotic cells. They lacked a cell nucleus, indicating the bacterial chromosome was not
surrounded by a membrane envelope. Therefore, bacteria have a prokaryotic (pro= primitive + karyon
= nucleus) type of cellular organization. Eubacteria and Archaea, thus, are prokaryotes.

The modern molecular biology era

By the 1970s, research on bacterial physiology, biochemistry and genetics had advanced to such an
extent that it was possible to experimentally manipulate the genetic material of living organisms.
With the invention of restriction enzymes, it became possible to introduce DNA from foreign sources
into bacteria and control its replication. This led to the development of fascinating field of
Biotechnology. In 1967, Carl Woese (Microfocus 1.9) originated the RNA World Hypothesis and
also discovered the extremophiles, Archaea. Prof Har Gobind Khorana (Microfocus 1.10) along with
Nirenberg and other coworker deciphered the genetic code and was awarded the Nobel Prize in
1968. Many diseases that were previously thought to have only behavioural or genetic components
have been found to involve microorganisms.

MICROFOCUS 1.9

Carl Woese, an American microbiologist, was born on July 15, 1928 in Syracuse,
New York. He is famous for defining the Archaea (a new domain or kingdom of
life) in 1977 by phylogenetic analysis of 16S ribosomal RNA, a technique pioneered
by Woese and which is now standard practice. He is also the originator of RNA
World Hypothesis in 1967, although not by name.

Carl Woese
 SCOPE AND HISTORICAL DEVELOPMENTS IN MICROBIOLOGY 19

Two Australians, Barry J. Marshall and Robin Warren won the 2005 Nobel Prize for showing
that bacterial infections of Helicobacter pylori (= Campylobacter pylori) and not the stress, is responsible
for painful ulcers in the stomach and intestine.The 1982 discovery transformed peptic ulcer disease
from a chronic, frequently disabling condition to one that can be cured by a short regimen of antibiotics
and medicines.
MICROFOCUS 1.10

Prof. Har Gobind Khorana was born on 2nd January, 1922 in Rajpura, Punjab,
India. He was awarded the Nobel Prize in Physiology/Medicine in 1968 for his
contribution to the elucidation of the genetic code. His research explained how
messages inscribed in genes are translated into proteins. He was also the first
person to successfully synthesize a gene in 1970. This achievement established
the foundation for the Biotechnology industry. The proteomics is defined as
where custom-designed genes are being widely used to engineer new plants and
animals.

Prof. H.G.

At the same time, nucleic acid sequencing methods were developed which left its impact in all
the areas of biology. Sequencing technology helped microbiologists to reveal phylogenetic
(evolutionary) relationships among prokaryotes, which led to evolutionary new concepts in the field
of biological classification. The field of Genomics is also a contribution of sequencing technology, in
which the comparative analysis of the genes of different organisms is carried out. The huge amounts
of genomic information now in hand are leading to major advances in medicine, microbial ecology,
industrial microbiology, and many other areas of biology. The genomics era has given birth to a new
subdiscipline, Proteomics. The proteomics is defined as the study of protein expression in cells. The
significance of such developments in molecular biology to all of biology is understood by the fact that
numerous Nobel Prizes have been awarded to researchers for their work in this field as shown in the
table 1.10.

Table 1.10: Nobel Laureates in Physiology or Medicine since 1901

Year Investigator(s) Discovery

1901 Emil von Behring Serum therapy, especially its application against diphtheria
1902 Ronald Ross Malaria, by which he has shown how it enters the organism
1903 Niels Ryberg Finsen Treatment of diseases, especially lupus vulgaris, with
concentrated light radiation
1904 Ivan Pavlov Physiology of digestion
1905 Robert Koch Investigations and discoveries in relation to tuberculosis
1906 Camillo Golgi and Santiago Structure of the nervous system
Ramony Cajal
1907 Alphonse Laveran Role played by protozoa in causing diseases
1908 Ilya Metchnikoff and Work on immunity
Paul Ehrlich

(Contd.)
20 A TEXTBOOK OF BASIC AND APPLIED MICROBIOLOGY

Year Investigator(s) Discovery

1909 Theodor Kocher Physiology, pathology and surgery of the thyroid gland
1910 Albrecht Kossel Cell chemistry, work on proteins, including the nucleic substances
1911 Allvar Gullstrand Dioptrics of the eye
1912 Alexis Carrel Vascular suture and the transplantation of blood vessels
and organs
1913 Charles Richet Anaphylaxis
1914 Robert Bárány Physiology and pathology of the vestibular apparatus
1919 Jules Bordet Discoveries relating to immunity
1920 August Krogh Capillary motor regulating mechanism
1922 Archibald V. Hill and Discovery relating to the production of heat in the muscle (Hill)
Otto Meyerhof and discovery of the fixed relationship between the consumption
of oxygen and the metabolism of lactic acid in the muscle
(Meyerhof)
1923 Frederick G. Banting Discovery of insulin
and John Macleod
1924 Willem Einthoven Mechanism of the Electrocardiogram
1926 Johannes Fibiger Discovery of the Spiroptera carcinoma
1927 Julius Wagner-Jauregg Therapeutic value of malaria inoculation in the treatment of
dementia paralytica
1928 Charles Nicolle Work on typhus
1929 Christiaan Eijkman and Discovery of the antineuritic vitamin (Eijkman) and discovery
Sir Frederick Hopkins of the growth stimulating vitamins (Hopkins)
1930 Karl Landsteiner Discovery of human blood groups
1931 Otto Warburg Nature and mode of action of the respiratory enzyme
1932 Sir Charles Sherrington Functions of neurons
and Edgar Adrian
1933 Thomas H. Morgan Role played by the chromosome in heredity
1934 George H. Whipple, Liver therapy in cases of anaemia
George R. Minot and
William P. Murphy
1935 Hans Spemann Organizer effect in embryonic development
1936 Sir Henry Dale and Chemical transmission of nerve impulses
Otto Loewi
1937 Albert Szent-Györgyi Biological combustion processes, with special reference to vitamin
C and the catalysis of fumaric acid
1938 Corneille Heymans Role played by the sinus and aortic mechanisms in the regulation
of respiration
1939 Gerhard Domagk Discovery of the antibacterial effect of prontosil
1943 Henrik Dam and Discovery of vitamin K and study on the chemical nature of
Edward A. Doisy vitamin K
(Contd.)
 SCOPE AND HISTORICAL DEVELOPMENTS IN MICROBIOLOGY 21

Year Investigator(s) Discovery

1944 Joseph Erlanger and Highly differentiated functions of single nerve fibres
Herbert S. Gasser
1945 Sir Alexander Fleming, Discovery of penicillin and its curative effect in various
Ernst B. Chain and infectious diseases
Sir Howard Florey
1946 Hermann J. Muller Production of mutations by means of X-ray irradiation
1947 Carl Cori, Gerty Cori Discovery of the course of the catalytic conversion of glycogen
and Bernardo Houssay (Cori and Cori) and discovery of the part played by the hormone
of the anterior pituitary lobe in the metabolism of sugar (Bernardo
Houssay)
1948 Paul Müller High efficiency of DDT as a contact poison against several
arthropods
1949 Walter Hess Discovery of the functional organization of the interbrain as a
and Egas Moniz coordinator of the activities of the internal organs (Walter Hess)
and discovery of the therapeutic value of leucotomy in certain
psychoses (Egas Moniz)
1950 Edward C. Kendall, Hormones of the adrenal cortex, their structure and biological
Tadeus Reichstein effects
and Philip S. Hench
1951 Max Theiler Yellow fever and how to combat it
1952 Selman A. Waksman Discovery of streptomycin, the first antibiotic effective against
tuberculosis
1953 Hans Krebs and Discovery of the citric acid cycle and discovery of co-enzyme
Fritz Lipmann A and its importance for intermediary metabolism
1954 John F. Enders, Thomas Ability of poliomyelitis viruses to grow in cultures of various
H. Weller and Frederick types of tissue
C. Robbins
1955 Hugo Theorell Nature and mode of action of oxidation enzymes
1956 André F. Cournand, Heart catheterization and pathological changes in the
Werner Forssmann and circulatory system
Dickinson W. Richards
1957 Daniel Bovet Discoveries relating to synthetic compounds that inhibit the
action of certain body substances, and especially their action
on the vascular system and the skeletal muscles
1958 George Beadle, Edward Genes act by regulating definite chemical events (Beadle and
Tatum, and Joshua Tatum) and discoveries concerning genetic recombination and
Lederberg the organization of the genetic material of bacteria (Lederberg)
1959 Severo Ochoa and Arthur Mechanisms in the biological synthesis of ribonucleic acid
Kornberg (RNA) and deoxyribonucleic acid (DNA)
1960 Sir Frank Macfarlane Acquired immunological tolerance
Burnet and Peter Medawar
1961 Georg von Békésy Physical mechanism of stimulation within the cochlea
(Contd.)
22 A TEXTBOOK OF BASIC AND APPLIED MICROBIOLOGY
Year Investigator(s)
1962 Francis Crick, James
Watson and Maurice Wilkins
1963 Sir John Eccles, Alan L.
Hodgkin and Andrew F.
Huxley
1964 Konrad Bloch and Feodor
Lynen
1965 François Jacob, André
L woff and Jacques Monod
1966 Peyton Rous and Charles
Brenton Huggins
1967 Ragnar Granit, Haldan K.
Hartline and George Wald
1968 Robert W. Holley, H. Gobind
Khorana and Marshall
W. Nirenberg
1969 Max Delbrück, Alfred D.
Hershey and Salvador
E. Luria
1970 Sir Bernard Katz, Ulf von
Euler and Julius Axelrod
1971 Earl W. Sutherland, Jr.
1972 Gerald M. Edelman and
Rodney R. Porter
1973 Karl von Frisch, Konrad
Lorenz and Nikolaas
Tinbergen
1974 Albert Claude, Christian
de Duve and George
E. Palade
1975 David Baltimore, Renato
Dulbecco, and Howard
M. Temin
1976 Baruch S. Blumberg and
D. Carleton Gajdusek
1977 Roger Guillemin, Andrew V.
Schally and Rosalyn Yalow
1978 Werner Arber,
Daniel Nathans and
Hamilton O. Smith
Discovery
Molecular structure of nucleic acids and its significance for
information transfer in living material
Ionic mechanisms involved in excitation and inhibition in the
peripheral and central portions of the nerve cell membrane
Mechanism and regulation of the cholesterol and fatty acid
metabolism
Genetic control of enzyme and virus synthesis
Discovery of tumour inducing viruses (Rous) and discoveries
concerning hormonal treatment of prostatic cancer (Huggins)
Primary physiological and chemical visual processes in the eye
Interpretation of the genetic code and its function in protein
synthesis
Replication mechanism and genetic structure of viruses
Humoral transmittors in the nerve terminals and the
mechanism for their storage, release and inactivation
Mechanisms of the action of hormones
Chemical structure of antibodies
Organization and elicitation of individual and social behaviour
patterns
Structural and functional organization of the cell
Interaction between tumour viruses and the genetic material
of the cell
New mechanisms for the origin and dissemination of
infectious diseases
Discoveries concerning the peptide hormone production of
the brain (Roger and Andrew) and for the development of
radioimmunoassays of peptide hormones (Rosalyn)
Discovery of restriction enzymes and their application to
problems of molecular genetics
(Contd.)
 SCOPE AND HISTORICAL DEVELOPMENTS IN MICROBIOLOGY 23

Year Investigator(s) Discovery

1979 Allan M. Cormack and Development of computer assisted tomography

Godfrey N. Hounsfield
1980 Baruj Benacerraf, Genetically determined structures on the cell surface that
Jean Dausset and regulate immunological reactions
George D. Snell
1981 Roger W. Sperry, David Discoveries concerning the functional specialization of the
H.Hubel, Torsten cerebral hemispheres (Roger) and for discoveries concerning
N. Wiesel information processing in the visual system (Hubel and Wiesel)
1982 Sune K. Bergström, Prostaglandins and related biologically active substances
Bengt I. Samuelsson and
John R. Vane
1983 Barbara McClintock Discovery of mobile genetic elements
1984 Niels K. Jerne, Georges Theories concerning the specificity in the development and
J.F. Köhler, control of the immune system and the discovery of the principle
César Milstein for production of monoclonal antibodies
1985 Michael S. Brown and Regulation of cholesterol metabolism
Joseph L. Goldstein
1986 Stanley Cohen and Discoveries of growth factors
Rita Levi-Montalcini
1987 Susumu Tonegawa Genetic principle for generation of antibody diversity
1988 Sir James W. Black, Important principles for drug treatment
Gertrude B. Elion and
George H. Hitchings
1989 J. Michael Bishop and Cellular origin of retroviral oncogenes
Harold E. Varmus
1990 Joseph E. Murray and Organ and cell transplantation in the treatment of human
E. Donnall Thomas disease
1991 Erwin Neher and Bert Function of single ion channel in cells
Sakmann
1992 Edmond H. Fischer and Reversible protein phosphorylation as a biological regulatory
Edwin G. Krebs mechanism
1993 Richard J. Roberts and Split genes
Phillip A. Sharp
1993 Kary Mullis Invention of the polymerase chain reaction (PCR)
1993 Hamilton Smith Specificity of action of restriction enzymes to splice foreign
components into DNA
1994 Alfred G. Gilman and G-proteins and their role in signal transduction in cell
Martin Rodbell
1995 Edward B. Lewis, Genetic control of early embryonic development
Christiane Nüsslein-
Volhard and
Eric F. Wieschaus
(Contd.)
24 A TEXTBOOK OF BASIC AND APPLIED MICROBIOLOGY

Year Investigator(s) Discovery

1996 Peter C. Doherty and Rolf Specificity of the cell mediated immune defence
M. Zinkernagel
1997 Stanley B. Prusiner Discovery of prions
1998 Robert F. Furchgott, Louis Nitric oxide as a signaling molecule in the cardiovascular system
J. Ignarro and Ferid Murad
1999 Günter Blobel Proteins have intrinsic signals that govern their transport
and localization in the cell
2000 Arvid Carlsson, Paul Signal transduction in the nervous system
Greengard and Eric
R. Kandel
2001 Leland H. Hartwell, Tim Key regulators of the cell cycle
Hunt and Sir Paul Nurse
2002 Sydney Brenner and Genetic regulation of organ development and programmed
H. Robert Horvitz and cell death
John E. Sulston
2003 Paul C. Lauterbur and Magnetic resonance imaging (MRI)
Sir Peter Mansfield
2004 Richard Axel and Linda Odorant receptors and the organization of the olfactory system
B. Buck
2005 Barry J. Marshall and Discovery of the bacterium Helicobacter pylori and its role in
J. Robin Warren gastritis and peptic ulcer disease
2006 Andrew Z. Fire RNA interference, gene silencing by double-stranded RNA
and Craig C. Mello
2007 Mario Capecchi, Oliver Gene targeting on knockout mouse using embryonic stem cells
Smithies and Martin Evans and in understanding gene disease relationship

Basic and applied sciences in microbiology

Microbiology, that has played a major role in the advancement of human health and welfare, is one
of the largest and most complex of the biological sciences as it deals with many diverse biological
disciplines. In addition to studying the natural history of microbes, it also deals with every aspect
of microbe-human and environmental interaction. These interactions include: ecology, genetics,
metabolism, infection, disease, chemotherapy, immunology, genetic engineering, industry and
agriculture. The branches that come under the large and expanding umbrella of microbiology are
categorized into basic and applied disciplines. The categorization is given below in the Table 1.11.

Table 1.11: Basic and applied disciplines in microbiology

Discipline Nature of study

A. Basic disciplines
Algology or Phycology Study of algae-simple aquatic organisms ranging from single-celled forms
to large seaweeds.
(Contd.)
 SCOPE AND HISTORICAL DEVELOPMENTS IN MICROBIOLOGY 25

Discipline Nature of study

Bacteriology Study of bacteria—the smallest, simplest, single-celled prokaryotic micro-

organisms and archaea – prokaryotic microorganisms which constitute an
ancient group intermediate between the bacteria and eukaryotes.
Mycology Study of fungi – microscopic eukaryotic forms (molds and yeasts), higher
forms (mushrooms, toadstools and puffballs), and slime molds.
Protozoology Study of protozoans—animal like and mostly single-celled, eukaryotic organisms.
Virology Study of viruses (infectious agents containing either DNA or RNA that require
living cells for their replication/ or reproduction) and viral diseases.
Parasitology Study of parasitism and parasites that include pathogenic protozoa, helminth
worms and some insects.
Microbial Ecology Study of interrelationships between microbes and environment.
Microbial Morphology Study of detailed structures of microorganisms.
Microbial Systematics Classification, naming, and identification of microorganisms and cons-
tructions of the phylogenetic tree of life.
Microbial Physiology Metabolism of microbes at the cellular and molecular levels.
Microbial Biochemistry Study of discovery of microbial enzymes and the chemical reactions carried
out by them.
Molecular Microbiology Study of genome (i.e., genomics) of microorganisms and construction of
phylogenetic tree based on rRNA.
Microbial Genetics Study of heredity and variation in varieties.
Molecular Biology The advanced study of the genetic material (DNA, RNA) and protein synthesis.
B. Applied disciplines
Immunology The immune system that protects against infections and attempts to under-
stand the many phenomena that are responsible for both acquired and
innate immunity, in addition to the study of antibody-antigen reactions in
the laboratory.
Agricultural Microbiology Study of relationships of microbes and crops with an emphasis on control
of plant diseases and improvement of yields.
Food Microbiology Interaction of microorganisms and food in relation to food bioprocessing,
food spoilage, food borne diseases and their prevention.
Dairy Microbiology Production of and maintenance in quality control of dairy products.
Industrial Microbiology Industrial uses of microbes in the production of alcoholic beverages,
vitamins, amino acids, enzymes, antibiotics and other drugs.
Marine Microbiology Study of microorganisms and their activity concerning human and animal
health in fresh, estuarine and marine waters.
Air Microbiology Role of aerospora in contamination and spoilage of food and dissemination
of plant and animal diseases through air.
Exomicrobiology Exploration for microbial life in outer space.
Diagnostic Microbiology Fundamental principles and techniques involved in the study of pathogenic
organisms as well as their application in the diagnosis of infectious diseases.
Epidemiology and Public Monitoring, control and spread of diseases in communities.
Health Microbiology
Biotechnology The scientific manipulation of living organisms, especially at the molecular
and genetic level to produce useful products.
26 A TEXTBOOK OF BASIC AND APPLIED MICROBIOLOGY

The new frontiers

The long span of four hundred and fifty years of microbiology has brought amazing insight into the
biology of microorganisms and has also brought with it new challenges, which have both positive
and negative effects upon the society. Diseases like AIDS, Bird’s flu and SARS seem to appear without
a trace and have challenged the basic understanding of microbial diseases. On the other hand, new
discoveries have opened a door for understanding how a cell works at the most fundamental level,
and newly discovered bacteria stretch the already overwhelming picture of microbial diversity.
Microbial ecology is providing new clues to the roles of microorganisms in the environment. Biofilms
are recognized as the dominant form of organization of microbial communities. The vast number of
unculturable microbes can be studied and characterized with genomic tools. The understanding of
microbial evolution has advanced with the use of genomic technologies and has provided new
perspectives on the relationships between microorganisms. Microorganisms play more positive roles
than simply causing infectious diseases. The majority of microbes are seen as rulers of the world
because of their essential and important beneficial roles that can provide humanity with an even
better and more healthful existence.

REVIEW QUESTIONS

1. Define microbiology. Enlist the various basic and applied areas of microbiology.
2. Why was the abandonment of the spontaneous generation theory so significant? Using the scientific
method, describe the steps you would take to test the theory of spontaneous generation.
3. Which early microbiologist was the most responsible for developing sterile laboratory techniques?
4. Which scientist is the most responsible for finally laying down the theory of spontaneous generation
to rest?
5. Enlist the contributions of Antony van Leeuwenhoek, Edward Jenner, Joseph Lister, Louis Pasteur,
Robert Koch and Joshua Lederberg.
6. What are the recent developments in the field of molecular microbiology?
7. List important commercial enzymes and their sources.
8. Name the scientists who first discovered Archaea?
9. What is a binomial system of nomenclature, and who proposed it?
10. Name the causative agents of: syphilis, whooping cough, blastomycosis, tinea cruris, toxoplasmosis,
giardiasis and schistosomiasis.
11. What are Koch’s postulates and how did they influence the development of microbiology?
12. How did Metchnikoff contribute to the development of immunology?
13. Describe the notable contributions of five scientists that resulted in the award of Nobel prizes to
them in microbiology.
14. How did Ferdinand Cohn and Carl Woese contribute to bacteriology and molecular biology respectively.
15. How did Pasteur’s Swan-neck experiment defeat the theory of spontaneous generation?
16. For what contributions are Hooke, Beijerinck, and Ehrlich remembered in microbiology?
17. How did the discovery of first antibiotic take place? Name the antibiotic and that mold from which
it was isolated.