Learning Resource Material
MODULE 01: INTRODUCTION TO GENERAL VETERINARY MICROBIOLOGY
Target Outcomes
At the end of the lesson, you are expected to:
1. Define what Microbiology is;
2. Identify the significant findings of the scientists behind the development of
Microbiology;
3. Explain the theories on the origin of diseases; and
4. Elucidate experiments that dispute the Abiogenesis Theory.
Abstraction
Microbiology is the study of living organisms of microscopic size (microorganisms), which
include bacteria, fungi, algae, protozoa and viruses.
Importance of microorganisms in our lives:
- they are important in the maintenance of an ecological balance on earth
- some live in humans and animals and are needed to maintain the animal’s health
- some are used to produce foods and chemicals
- some cause disease
DISCOVERY OF THE MICROBIAL WORLD
Prior to the 16th century, most people (including scientists) thought that disease was caused
by one of a variety of things:
• An imbalance of the body's humors (fluids), leading to sickness (hence cupping and
bleeding)
• Miasma (bad air or "vapors", hence "influenza")
• A curse from God or witchcraft
• Possession by demons or other spiritual forces
Even before microorganisms were seen, some investigators suspected their existence and
responsibility for disease. Among others, the Roman philosopher Lucretius (about 98-55 B.C.)
and the physician Girolamo Fracastoro (1478-1553) suggested that disease was caused by
invisible living creatures.
1589 Thomas Moffet – the vectors of disease began to be identified, and a variety of infections
were identified as being caused by fungal invasion.
Plenciz – stated that living agents are the cause of diseases and that different germs were
responsible for different diseases.
The earliest microscopic observations appear to have been made between 1625 and 1630
on bees and weevils by the Italian Francesco Stelluti, using a microscope supplied by Galileo.
About 1674, the invisible world of microorganisms was discovered by the Dutch merchant Anton
van Leeuwenhoek (1623-1723).
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1665 Robert Hooke (English) – first to use the word cell; he observed the honeycomb-like
structure in a thin slice of cork.
1673 Antoni van Leeuwenhoek (Dutch) – first to observe, using home-ground lenses, and
accurately record and report microorganisms (which he referred to as “animalcules”).
1838-1839 Matthias Schleiden and Theodore Schwann (Germans) – credited with the cell theory
(concept that all living organisms are made up of cells); they recognized that all cells from
any organism are similar in structure.
CONTROVERSY OVER SPONTANEOUS GENERATION
Spontaneous Generation – the idea that living organisms could arise from nonliving matter.
Leeuwenhoek’s observations, though their importance was not fully recognized until
nearly 200 years later, came at a time when the theory of spontaneous generation was being
challenged. This theory had been advanced to explain the occurrence of flies on putrefying
meat, mice in decomposing fodder, and snakes in stagnant water. To disprove these ideas, it
was necessary to carry out carefully controlled experiments.
1626-1697 Francesco Redi (Italian) – demonstrated in experiments using covered and uncovered
jars that maggots appear on decaying meat only when flies are able to lay eggs on the meat.
The results were a serious blow to the long-held belief on spontaneous generation.
1713-1781 John Needham – believer of spontaneous generation. He claimed in a paper published
in 1749 that even after he heated nutrient fluids (chicken broth and corn infusions) before
putting them into covered flasks, the cooled solutions were soon teeming with microbes. He
thought organic matter contained a vital force that could confer the properties of life on
nonliving matter.
1729-1799 Lazzaro Spallanzani– improved on Redi’s experimental design by first sealing the
glass flasks that contained water and seeds. If the sealed flasks were placed in a boiling
water for ¾ of an hour, no growth took place as long as the flasks remained sealed. He
proposed that air carried germs to the culture medium, but also commented that the external
air might be required for growth of animals already in the medium. The supporters of
spontaneous generation maintained that heating the air in sealed flasks destroyed its ability
to support life.
1815-1873 Franz Schultze – passed air through strong acid solutions into boiled infusions.
Microbes did not appear. His results supported Spallanzani’s observations.
1810-1882 Theodor Schwann – passed air into his flasks through red-hot tubes. Microbes did
not appear, too. His results also supported Spallanzani’s observations.
1850 Georg Friedrich Schröder and Theodor von Dusch – passed air through cotton stoppers into
flasks; the microbes were filtered by the cotton fibers so that growth did not occur even
though the air had not been heated.
1858 Rudolf Virchow (German) – challenged spontaneous generation with the concept of
biogenesis. An extension of the cell theory, biogenesis claims that living cells can arise
only from preexisting living cells).
1859 Felix-Archimede Pouchet – revived the concept of S.G., published reports “proving” its
occurrence.
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EXPERIMENTS TO DISPROVE SPONTANEOUS GENERATION
As mentioned above Redi, Spallanzani, Schultze, Schwann, Schröder and von Dusch made
experiments with results against spontaneous generation. Aside from them Pasteur and Tyndall
also made experiments to finally disprove spontaneous generation.
1861 Louis Pasteur (French) – resolved the issue on spontaneous generation with a series of
ingenious and persuasive experiments. He first filtered air through cotton and found that
objects resembling plant spores had been trapped. If a piece of the cotton was placed in
sterile medium after air had been filtered through it, microbial growth appeared. Next he
placed nutrient solutions in flasks, heated the necks in flame, and drew them out into a
variety of curves (swan-neck shape), while keeping the ends of the necks open to the atmosphere.
Pasteur then boiled the solutions for a few minutes and allowed them to cool. No growth took
place even though the contents of the flasks were exposed to air. Pasteur pointed out
that no growth occurred because dust and germs had been trapped on the walls of the curved
necks. If the necks were broken, growth commenced immediately. He further demonstrated that
if he placed a series of these flasks along a busy road, opened them and resealed them a few
minutes later, microorganisms would grow in nearly all the flasks. On the other hand, if he
performed the same experiment on top of a mountain where there is little activity, practically
none of his flasks became contaminated.
1879 Thomas Huxley – gave his "Biogenesis and Abiogenesis" lecture. The speech offered
powerful support for Pasteur's claim to have experimentally disproved spontaneous generation.
1877 John Tyndall – he dealt a final blow to spontaneous generation through his specially
designed box. He observed that when one shines a bright beam of light through the air, the
pathway of light can be seen because it is refracted by the dust particles in the air. If
however, the air were 100% free of dust, one could not see the beam.
After the box had settled so that he can no longer see a beam of light pass through
the air, he carefully placed a box of sterile infusions in the box. As long as the air was
not disturbed, the infusions remained sterile even though they were open to air. This
demonstrated that microorganisms existed in dust particles in the air and that they were not
spontaneously generated in hay or in meat infusions.
Tyndall’s work resulted in a second interesting observation. He noted that on some
occasions, only a few minutes of boiling would sterilize his meat infusions, whereas at other
times even 5 ½ hours of boiling was not sufficient. He eventually recognized that there were
heat-stable microorganisms know as endospores. He found that if an infusion was heated to
boiling for 10 minutes, the heat-labile forms known as vegetative cells were destroyed. If
this infusion was then allowed to remain at room temperature overnight, most of the endospores
would germinate to form vegetative cells that are easy to kill.
ROLE OF MICROORGANISMS IN DISEASE CAUSATION
1796 Edward Jenner – inoculated an 8-yr. old boy named James Phipps with cowpox, then 8 weeks
later inoculated him with pus from a smallpox lesion. The boy showed no effects of smallpox
and Jenner repeated the experiment.
1805-1885 Jacob Henle (Friedrich Gustav Jakob Henle) – established principles for the Germ
Theory of Disease. He developed the concepts of contagium vivum and contagium animatum,
respectively - thereby following ideas of Fracastoro and the work of Bassi; thus co-founding
the theory of microorganisms as the cause of infective diseases. He did not find a special
species of bacteria himself - this was achieved by his student Robert Koch. These two put up
the fundamental rules of cleanly defining disease-causing microbes: the Henle-Koch postulates
(more popularly known as Koch postulates only).
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1834 Agostino Bassi – made the first specific causal relationship between microorganism and
disease by proving that a fungus infection caused a silkworm disease.
1809-1894 Oliver Wendell Holmes (American) – stressed contagiousness of puerperal fever; that
agent was carried from one mother to another by doctors.
1840’s Ignaz Philipp Semmelweis (Hungarian) – He showed that hand washing between visiting
mothers can prevent childbirth fever. He pioneered the use of antiseptics in obstetrical
practice; death due to perinatal infections were minimized.
1854 John Snow (English) - Dr. John Snow studied a cholera outbreak in the Soho neighborhood
of London and determined it was caused by contaminated water at the Broad Street pump. His
methods found the field of epidemiology.
1865 Louis Pasteur – the “Grandfather of Microbiology.” He showed that a protozoan caused
pebrine, another silkworm disease that was destroying the silk industry in France. He
established criteria whereby infected silkworm moths could be recognized microscopically.
Therefore, when females free of infection were used, the silkworms remained free of disease.
It is Pasteur who is considered to be the father of germ theory, bringing the theory of
disease away from superstition and folklore into a more scientific arena.
1867 Joseph Lister (English) – he was credited with the first attempt to prevent postsurgical
infection by the use of aseptic technique by using diluted phenol both in wound dressings
and as an aerosol during surgical procedures.
1820-1910 Florence Nightingale – organized hospitals which minimized cross-infection.
1873 Gerhard Henrik Armauer Hansen – discovered the leprosy bacillus (Mycobacterium leprae)
and demonstrated that leprosy is a contagious disease and not inherited as was the popular
belief. In many countries leprosy is still called Hansen's disease in his honor.
1875 Ferdinand Cohn - effectively founded the science of bacteriology (a branch of
microbiology which studies bacteria). His main contribution was the classification of
bacteria, and he coined the term Bacillus.
1876 Robert Koch (German) – discovered that anthrax was caused by a typical bacterium Bacillus
anthracis. In 1882, he isolated the tuberculosis bacillus, Mycobacterium tuberculosis.He
established a sequence of experimental steps to directly relate a specific microbe to a
specific disease, called “Koch’s postulates” in 1884.
Koch’s Postulates
1. The microorganism must be found in all organisms suffering from the disease, but not
in healthy organisms.
2. The microorganism must be isolated from a diseased organism and grown in pure
culture.
3. The cultured microorganism should cause disease when introduced into a healthy
organism.
4. The microorganism must be reisolated from the inoculated, diseased experimental host
and identified as being identical to the original specific causative agent.
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Koch also developed techniques to grow
bacteria for study, and further developed
methods of controlling the spread of
bacteria through sterilisation. The field
of bacteriology was born.
Koch developed the technique of growing
bacteria and observed individual colonies
of identical, pure cells.
1850-1934 Frannie Eilshemius Hesse – she
was wife of Koch’s assistant, Walther Hesse.
She was the first to propose the use of
agar to solidify a nutrient medium. She discovered that agar was a good solidifying agent
because agar will melt only at about the temperature of boiling water and will not resolidify
until it is cooled to approximately 43°C. Her discovery proved to be a major advance in
bacteriologic technique and silenced critics who strongly proclaimed that there was only one
species of bacteria that might assume different shapes and forms.
Richard Petri – one of Koch’s assistants, he developed the Petri dish (plate), a container
for solid culture media. These developments made possible the isolation of pure cultures
that contained only one type of bacterium, and directly stimulated progress in all areas of
bacteriology.
1886 John Brown Buist – was the first person to see a virus.
1892 Dmitri Ivanowski – published the first evidence of the filterability of a pathogenic
agent, the virus of tobacco mosaic disease.
1899 Martinus Beijerinck – recognized the unique nature of Ivanowski's discovery. He coins
the term contagium vivum fluidum - a contagious living fluid.
1899 Friederich Loeffler and Paul Frosch – discovered that foot and mouth disease was also
caused by a filterable agent.
1915-1917 Frederick Twort and Felix d'Herelle – discovered bacterial viruses. d'Herelle
coined the term bacteriophage (devourer of bacteria).
1918 In the fall of 1918, as World War I was ending, an influenza pandemic of unprecedented
virulence swept the globe, leaving some 40 million dead in its wake. A search for the
responsible agent began in earnest that year, leading to the first isolation of an influenza
virus by 1930.
1957 D. Carleton Gajdusek – proposed that a slow virus was responsible for the wasting disease
kuru. In subsequent years several diseases are shown to be caused by slow viruses (later
renamed prions) including mad cow disease.
*Some Famous Microbiologists
and their Breakthroughs*
BACTERIA & ARCHAEA
Ferdinand J Cohn published an early classification of bacteria (genus name Bacillus) for
the first time in 1875.
Ilya Ilich Metchnikoff received the Nobel Prize in 1908 with Ehrlich, for demonstrating
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phagocytosis - the consumption of foreign particles and bacteria by the body's own
antibodies.
Alice Catherine Evans (1881-1975). Her work in Wisconsin Dept. of Agriculture led to the
identification of bacteria in fresh milk. Her later research, at the National Institutes
of Health (NIH), improved the treatment of epidemic meningitis and she became first female
president of the American Society for Microbiology in 1928.
Rebecca Craighill Lancefield (1895-1981). Developed a system of classification for Group
A streptococcal bacteria - the Lancefield Grouping - which identifies bacteria including
those causing scarlet fever, sore throat and erysipelas. She received the Lasker Award
and was elected to the National Academy of Sciences.
Holger Jannasch was one of the world's leading experts on life around mid-ocean
hydrothermal vents. His team discovered Pyrolobus fumarii, an Archaea, at the mid-
Atlantic ridge in 1996. Holger died in 1999.
IMMUNIZATION & TREATMENT
Louis Pasteur (1822 -1895). Developed a method of immunizing against a disease (chicken
cholera) using a weakened (attenuated) strain of the pathogen in 1880. In 1885 he carried
out successful, but unethical, experiments with rabies on a child. The term virus
(poison) was coined by Pasteur.
Emil von Behring received the Nobel Prize in 1901 for his work with Shibasaburo Kitasato
on the antitoxin serum for diptheria.
Paul Ehrlich in 1912, announced the discovery of an effective cure for syphilis, the first
chemotherapeutic agent for a bacterial disease.
Margaret Pittman (1901-1995). Identified the cause of whooping cough, which led to the
development of an improved vaccine. She became the first woman to direct a laboratory at
the NIH and was cholera consultant to the World Health Organization and a leader in the
standardization of vaccines.
Gerhard J Domagk used a chemically-produced antimetabolite to kill streptococci in mice,
in 1935. It was later used on human patients and he received the 1939 Nobel Prize for his
work.
Alexander Fleming discovered penicillin in 1928 whilst working at St Mary's Hospital in
London and published the first paper on it the following year. He received the Nobel
Prize in 1945, with Howard Florey and Ernst Chain, for their work on Penicillium notatum.
William A Hinton (1883-1959). Directed the Massachusetts State Wasserman Laboratory from
1915 and taught for 30 years at Harvard University Medical School, becoming full professor
there in 1949. He developed a widely-used test for diagnosing syphilis. He was
instrumental in establishing the Eisenhower Scholarship at Harvard University.
Albert Shatz, E Bugie and Selman Waksman discovered streptomycin in 1944, which was then
used to counter tuberculosis. Selman Waksman received the Nobel Prize in 1952.
PLANTS & SOIL
Sergei Winogradsky, in 1890, isolated nitrifying bacteria in soil and described the
organisms which are responsible for nitrification.
Dmitri Ivanowski published the first evidence of tobacco mosaic virus, in 1892.
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CB Van Niel, by his work on photosynthetic bacteria, in 1931 explained the fixation of
carbon-dioxide in plants and suggested that plants use water as a source of electrons and
release oxygen.
Wendell Stanley, in 1935, demonstrated the tobacco mosaic virus remains active even after
crystallization. He received the Nobel Prize in 1946 with Northrop and Sumner.
VECTORS
Theobald Smith and F L Kilbourne, in 1893 provided evidence of a zoonotic disease (in this
case animal host and arthropod vector) by establishing that ticks carry Babesia microti.
Walter Reed worked on the viral agent for yellow fever being transmitted to humans by
mosquitoes, which inspired mosquito eradication and the Yellow Fever Commission in 1900.
VIRUSES & PRIONS
Frederick Twort, between 1915 and 1917, first discovered a bacterial virus which was also
independently described and named as a bacteriophage by Felix d'Herrelle.
Francis Peyton Rous was awarded the 1966 Nobel Prize for work he carried out on chickens
in 1911, that gave the first experimental proof of a virus causing cancer.
Stanley Prusiner found evidence in 1982 that a class of infections he called "prions"
cause scrapie, a fatal neurodegenerative disease of sheep and was awarded the Nobel Prize
in 1997.
Luc Montaigner and Robert Gallo announced in 1983 the discovery of the human
immunodeficiency virus (HIV) believed to cause AIDS.
CELLS & CULTURES
Joseph Lister published his study of lactic fermentation of milk in 1878, using a method
of isolating a pure culture of the bacterium responsible.
Martinus Beijerinck developed an enrichment culture to create the best conditions for
growth of required bacterium in 1889. Whilst working on tobacco mosaic virus in 1899, he
discovered that a filtrate free of bacteria can still transmit the disease, by some other
agent.
Robert Koch published a paper on the bacterium which causes anthrax in 1876. In 1881 he
developed the use of gelatin on glass plates as a means for culturing bacteria colonies
for experiments. He was awarded the Nobel Prize in 1905 for his work on the Tubercule
bacillus of tuberculosis.
Albert Jan Kluyver and Hendrick Jean Louis Donker in 1926 proposed a model suitable for
aerobic and anaerobic organisms, for metabolism in cells based on the transfer of hydrogen
atoms.
John Franklin Enders, Thomas H Weller and Frederick Chapman Robbins were awarded the Nobel
Prize in 1954 for developing a technique to grow the poliovirus in test tube cultures of
human tissue, thus enabling the isolation and study of viruses in the laboratory.
Peter Mitchell proposed the chemiosmotic theory in 1959, which explains ATP synthesis,
solute accumulations/expulsions, and cell movement. He was awarded the Nobel Prize in
1978.
George Kohler and Cesar Milstein in 1975, produced specific antibodies that can survive
indefinitely in tissue culture, which can then be used for diagnostic tests and to study
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cell function. With Jerne, they were awarded the Nobel Prize in 1984.
GENETICS
Frederick Griffith discovered transformation in bacteria in 1928 and established the
foundation of molecular genetics.
Oswald Avery, Colin MacLeod and Maclyn McCarty in 1944 took up Griffith's work and showed
that Streptococcus pneumoniae could transform from an avirulent to a virulent phenotype by
the transfer of DNA.
George Beadle and Edward Tatum published a paper in 1941 on fungus experiments, which
demonstrated that specific genes are expressed through action of designated enzymes the
"one gene - one enzyme" concept. They were awarded the Nobel Prize with Lederberg in
1958.
Joshua Lederberg and Edward Tatum published the first paper on conjugation in bacteria in
1946. Joshua Lederberg and Norton Zinder showed that a phage of Salmonella typhimurium
can carry DNA from one bacterium to another and reported on transduction (transfer of
genetic information by viruses) in 1952.
Alfred Hershey and Martha Chase suggested in 1952 that only DNA is required for viral
replication, after using radioactive isotopes to track protein and DNA.
Max Delbruck and Salvador Luria, demonstrated statistically in 1943 that inheritance in
bacteria follow Darwin's principles and that mutant bacteria occurring randomly can still
bestow viral resistance without the virus being present. They received the Nobel Prize
with Hershey in 1969.
Sydney Brenner, Francois Jacob and Matthew Meselson used phage-infected bacteria to
confirm the existence of messenger RNA in 1961.
Francis Crick, Maurice Wilkins and James Watson were awarded the Nobel Prize in 1962 for
describing the double-helix structure of DNA. This was based on the X-ray crystallography
of DNA done by Rosalind Franklin, who had died of cancer four years earlier.
Charles Yanofsky and colleagues in 1964, defined the relationship between the order of
mutatable sites in the gene coding for Escherichia coli.
Francois Jacob and Jacques Monod (together with David Perrin and Carmen Sanchez) proposed
the operon concept for control of bacteria gene action. Jacob, Monod and Lwoff were
awarded the Nobel Prize in 1965.
Marsha Nirenberg and J H Matthaei realized in 1961 that the triplet UUU must code for
phenylalanine and thus started to decipher the genetic code. Nirenberg, Robert Holley and
Har Gorbind Khorana were awarded the Nobel Prize in 1968.
Stanley Cohen, Annie Chang, Robert Helling and Herbert Boyer in 1973, showed that
recombinant DNA molecules will reproduce if inserted into bacteria cells, this paved the
way for gene modification and cloning.
Howard Temin and David Baltimore independently discovered reverse transcriptase in RNA
viruses in 1970, establishing a pathway for genetic information flow from RNA to
DNA. With Dulbecco they were awarded the Nobel Prize in 1975.
Carl Woese in 1977, used ribosomal RNA analysis to identify Archaea whose genetic makeup
is distinct from, but related to, both Bacteria and Eucarya.
Frederick Sanger was the first British scientist to be awarded two Nobel Prizes. He
received his first Nobel Prize in 1958 for discovering the sequence of amino acids in the
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hormone insulin. Sanger, Walter Gilbert and Berg received the Nobel Prize in 1980 for
their work on the chemical structure of genes.
Kary Mullis used a heat-stable enzyme to establish Polymerase Chain Reaction (PCR)
technology and amplify target DNA in 1986. He received the Nobel Prize in 1993.
Craig Venter, Hamilton Smith, Claire Fraser and colleagues determined the first complete
genome sequence of a microorganism - Haemophilus influenzae RD, in 1995.
Utilization of Learning
Self-gauging assessment:
1. Define Microbiology.
2. What is the Cell Theory?
3. Who introduced the use of cotton plug?
4. Who developed the first rabies vaccine?
5. Who introduced the use of Polymerase Chain Reaction to identify microorganism?
IMPLICATIONS:
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