Medina College

College of Pharmacy

1st Lab Assignment

In partial fulfillment

of the requirements in

Micro 1 (lab) – Pharmaceutical Microbiology & Parasitology (lab)

Submitted to:

Mr. Mark B. Quiamco, RPh

[ 14 June 2019 ]

Submitted by:

Roa, Josefina Carmen P.

 I. Brief History of Microbiology
Microbiology has had a long, rich history, initially centered in the causes of infectious diseases but
now including practical applications of the science. Many individuals have made significant contributions
to the development of microbiology.

Early history of microbiology. Historians are unsure who made the first observations of
microorganisms, but the microscope was available during the mid‐1600s, and an English scientist
named Robert Hooke made key observations. He is reputed to have observed strands of fungi among
the specimens of cells he viewed. In the 1670s and the decades thereafter, a Dutch merchant
named Anton van Leeuwenhoek made careful observations of microscopic organisms, which he
called animalcules. Until his death in 1723, van Leeuwenhoek revealed the microscopic world to
scientists of the day and is regarded as one of the first to provide accurate descriptions of protozoa, fungi,
and bacteria.

After van Leeuwenhoek died, the study of microbiology did not develop rapidly because
microscopes were rare and the interest in microorganisms was not high. In those years, scientists
debated the theory of spontaneous generation, which stated that microorganisms arise from lifeless
matter such as beef broth. This theory was disputed by Francesco Redi, who showed that fly maggots do
not arise from decaying meat (as others believed) if the meat is covered to prevent the entry of flies. An
English cleric named John Needhamadvanced spontaneous generation, but Lazzaro
Spallanzani disputed the theory by showing that boiled broth would not give rise to microscopic forms of
life.

Louis Pasteur and the germ theory. Louis Pasteur worked in the middle and late 1800s. He
performed numerous experiments to discover why wine and dairy products became sour, and he found
that bacteria were to blame. Pasteur called attention to the importance of microorganisms in everyday life
and stirred scientists to think that if bacteria could make the wine “sick,” then perhaps they could cause
human illness.

Pasteur had to disprove spontaneous generation to sustain his theory, and he therefore devised a
series of swan‐necked flasks filled with broth. He left the flasks of broth open to the air, but the flasks
had a curve in the neck so that microorganisms would fall into the neck, not the broth. The flasks did not
become contaminated (as he predicted they would not), and Pasteur's experiments put to rest the notion
of spontaneous generation. His work also encouraged the belief that microorganisms were in the air and
could cause disease. Pasteur postulated the germ theory of disease, which states that microorganisms
are the causes of infectious disease.

Pasteur's attempts to prove the germ theory were unsuccessful. However, the German
scientist Robert Koch provided the proof by cultivating anthrax bacteria apart from any other type of
organism. He then injected pure cultures of the bacilli into mice and showed that the bacilli invariably
caused anthrax. The procedures used by Koch came to be known as Koch's postulates (Figure ). They
provided a set of principles whereby other microorganisms could be related to other diseases.
The development of microbiology. In the late 1800s and for the first decade of the 1900s,
scientists seized the opportunity to further develop the germ theory of disease as enunciated by Pasteur
and proved by Koch. There emerged a Golden Age of Microbiology during which many agents of
different infectious diseases were identified. Many of the etiologic agents of microbial disease were
discovered during that period, leading to the ability to halt epidemics by interrupting the spread of
microorganisms.

Despite the advances in microbiology, it was rarely possible to render life‐saving therapy to an
infected patient. Then, after World War II, the antibiotics were introduced to medicine. The incidence of
pneumonia, tuberculosis, meningitis, syphilis, and many other diseases declined with the use of antibiotics.
 Work with viruses could not be effectively performed until instruments were developed to help
scientists see these disease agents. In the 1940s, the electron microscope was developed and
perfected. In that decade, cultivation methods for viruses were also introduced, and the knowledge of
viruses developed rapidly. With the development of vaccines in the 1950s and 1960s, such viral diseases
as polio, measles, mumps, and rubella came under control.

Modern microbiology. Modern microbiology reaches into many fields of human endeavor,
including the development of pharmaceutical products, the use of quality‐control methods in food and
dairy product production, the control of disease‐causing microorganisms in consumable waters, and the
industrial applications of microorganisms. Microorganisms are used to produce vitamins, amino acids,
enzymes, and growth supplements. They manufacture many foods, including fermented dairy products
(sour cream, yogurt, and buttermilk), as well as other fermented foods such as pickles, sauerkraut, breads,
and alcoholic beverages.
One of the major areas of applied microbiology is biotechnology. In this discipline,
microorganisms are used as living factories to produce pharmaceuticals that otherwise could not be
manufactured. These substances include the human hormone insulin, the antiviral substance interferon,
numerous blood‐clotting factors and clotdissolving enzymes, and a number of vaccines. Bacteria can be
reengineered to increase plant resistance to insects and frost, and biotechnology will represent a major
application of microorganisms in the next century.

The steps of Koch's postulates used to relate a specific microorganism to a specific disease. (a)
Microorganisms are observed in a sick animal and (b) cultivated in the lab. (c) The organisms are injected
into a healthy animal, and (d) the animal develops the disease. (e) The organisms are observed in the
sick animal and (f) reisolated in the lab.

The development of microbiology. In the late 1800s and for the first decade of the 1900s,
scientists seized the opportunity to further develop the germ theory of disease as enunciated by Pasteur
and proved by Koch. There emerged a Golden Age of Microbiology during which many agents of
different infectious diseases were identified. Many of the etiologic agents of microbial disease were
discovered during that period, leading to the ability to halt epidemics by interrupting the spread of
microorganisms.
 Despite the advances in microbiology, it was rarely possible to render life-saving therapy to an
infected patient. Then, after World War II, the antibiotics were introduced to medicine. The incidence of
pneumonia, tuberculosis, meningitis, syphilis, and many other diseases declined with the use of antibiotics.

Work with viruses could not be effectively performed until instruments were developed to help
scientists see these disease agents. In the 1940s, the electron microscope was developed and
perfected. In that decade, cultivation methods for viruses were also introduced, and the knowledge of
viruses developed rapidly. With the development of vaccines in the 1950s and 1960s, such viral diseases
as polio, measles, mumps, and rubella came under control.

II. Bacterial Classifications/Shapes

a) Bacillus: These are rod-shaped or filament-shaped bacteria. They are of four types like
i) Monobacillus: This is a single rod-shaped bacillus bacteria.
ii) Diplobaciullus: These are a pair of rod-shaped bacteria. Two bacteria cells stick together.
They can also be present as four-celled as a tetrad.
iii) Streptobacilli: This is a chain of rod-shaped bacteria. Bacilli bacteria arranged like a long
chain.
iv) Palisade: Here two cells of Bacillus are arranged side by side like sticks in a matchbox

Bacillus is rod-shaped, cocci

are spherical, cholera
bacteria is comma-shaped
and syphilis bacteria is spiral
shaped. Further, cocci and
bacilli can be in groups or
chains.
 b) Coccus: These are spherical shaped bacteria or oval shaped. Based on the number and
their arrangement they are divided
i) Monococcus which is a single-celled round-shaped bacteria.
ii) Diplococci is two spherical shaped bacteria existing as pairs.
iii) Streptococci is a chain of many round-shaped bacteria.
iv) Staphylococci is a group of spherical bacteria arranged like a bunch of grapes.
v) Sarcina is a type where 8 round shaped bacteria are arranged in cubical shape.
c) Comma-shaped bacteria: Here is the bacteria is slightly bent and looks like a comma.
Ex: Vibrio cholera bacteria causing cholera.
d) Spirillum bacteria: This is a long spiral-shaped bacteria. They are also called as
spirochetes. These are spiral or hair like in shape. Ex: syphilis-causing bacteria.
e) Pleomorphism: Though most bacteria have a specific shape, some do not. They exist in
multiple shapes. Examples include Acetobacter.

III. Bacterial Cell Structure

 Capsule - Some species of bacteria have a third protective covering, a capsule made up of
polysaccharides (complex carbohydrates). Capsules play a number of roles, but the most
important are to keep the bacterium from drying out and to protect it from phagocytosis (engulfing)
by larger microorganisms. The capsule is a major virulence factor in the major disease-causing
bacteria, such as Escherichia coli and Streptococcus pneumoniae. Nonencapsulated mutants
of these organisms are avirulent, i.e. they don't cause disease.
 Cell Envelope - The cell envelope is made up of two to three layers: the interior cytoplasmic
membrane, the cell wall, and -- in some species of bacteria -- an outer capsule.
 Cell Wall - Each bacterium is enclosed by a rigid cell wall composed of peptidoglycan, a protein-
sugar (polysaccharide) molecule. The wall gives the cell its shape and surrounds the cytoplasmic
membrane, protecting it from the environment. It also helps to anchor appendages like the pili and
flagella, which originate in the cytoplasm membrane and protrude through the wall to the outside.
The strength of the wall is responsible for keeping the cell from bursting when there are large
differences in osmotic pressure between the cytoplasm and the environment.

Cell wall composition varies widely amongst bacteria and is one of the most important factors in
bacterial species analysis and differentiation. For example, a relatively thick, meshlike structure
that makes it possible to distinguish two basic types of bacteria. A technique devised by Danish
physician Hans Christian Gram in 1884, uses a staining and washing technique to differentiate
between the two forms. When exposed to a gram stain, gram-positive bacteria retain the purple
color of the stain because the structure of their cell walls traps the dye. In gram-negative bacteria,
the cell wall is thin and releases the dye readily when washed with an alcohol or acetone solution.

 Cytoplasm - The cytoplasm, or protoplasm, of bacterial cells is where the functions for cell growth,
metabolism, and replication are carried out. It is a gel-like matrix composed of water, enzymes,
nutrients, wastes, and gases and contains cell structures such as ribosomes, a chromosome, and
plasmids. The cell envelope encases the cytoplasm and all its components. Unlike the eukaryotic
(true) cells, bacteria do not have a membrane enclosed nucleus. The chromosome, a single,
continuous strand of DNA, is localized, but not contained, in a region of the cell called the nucleoid.
All the other cellular components are scattered throughout the cytoplasm.

One of those components, plasmids, are small, extrachromosomal genetic structures carried by
many strains of bacteria. Like the chromosome, plasmids are made of a circular piece of DNA.
Unlike the chromosome, they are not involved in reproduction. Only the chromosome has the
genetic instructions for initiating and carrying out cell division, or binary fission, the primary means
of reproduction in bacteria. Plasmids replicate independently of the chromosome and, while not
essential for survival, appear to give bacteria a selective advantage.

Plasmids are passed on to other bacteria through two means. For most plasmid types, copies in
the cytoplasm are passed on to daughter cells during binary fission. Other types of plasmids,
however, form a tubelike structure at the surface called a pilus that passes copies of the plasmid to
other bacteria during conjugation, a process by which bacteria exchange genetic information.
Plasmids have been shown to be instrumental in the transmission of special properties, such as
antibiotic drug resistance, resistance to heavy metals, and virulence factors necessary for infection
of animal or plant hosts. The ability to insert specific genes into plasmids have made them
extremely useful tools in the fields of molecular biology and genetics, specifically in the area of
genetic engineering.

 Cytoplasmic Membrane - A layer of phospholipids and proteins, called the cytoplasmic

membrane, encloses the interior of the bacterium, regulating the flow of materials in and out of the
cell. This is a structural trait bacteria share with all other living cells; a barrier that allows them to
selectively interact with their environment. Membranes are highly organized and asymmetric
having two sides, each side with a different surface and different functions. Membranes are also
dynamic, constantly adapting to different conditions.

 Flagella - Flagella (singular, flagellum) are hairlike structures that provide a means of locomotion
for those bacteria that have them. They can be found at either or both ends of a bacterium or all
over its surface. The flagella beat in a propeller-like motion to help the bacterium move toward
nutrients; away from toxic chemicals; or, in the case of the photosynthetic cyanobacteria; toward
the light.
  Nucleoid - The nucleoid is a region of cytoplasm where the chromosomal DNA is located. It is not
a membrane bound nucleus, but simply an area of the cytoplasm where the strands of DNA are
found. Most bacteria have a single, circular chromosome that is responsible for replication,
although a few species do have two or more. Smaller circular auxiliary DNA strands, called
plasmids, are also found in the cytoplasm.

 Pili - Many species of bacteria have pili (singular, pilus), small hairlike projections emerging from
the outside cell surface. These outgrowths assist the bacteria in attaching to other cells and
surfaces, such as teeth, intestines, and rocks. Without pili, many disease-causing bacteria lose
their ability to infect because they're unable to attach to host tissue. Specialized pili are used for
conjugation, during which two bacteria exchange fragments of plasmid DNA.

 Ribosomes - Ribosomes are microscopic "factories" found in all cells, including bacteria. They
translate the genetic code from the molecular language of nucleic acid to that of amino acids—the
building blocks of proteins. Proteins are the molecules that perform all the functions of cells and
living organisms. Bacterial ribosomes are similar to those of eukaryotes, but are smaller and have
a slightly different composition and molecular structure. Bacterial ribosomes are never bound to
other organelles as they sometimes are (bound to the endoplasmic reticulum) in eukaryotes, but
are free-standing structures distributed throughout the cytoplasm. There are sufficient differences
between bacterial ribosomes and eukaryotic ribosomes that some antibiotics will inhibit the
functioning of bacterial ribosomes, but not a eukaryote's, thus killing bacteria but not the eukaryotic
organisms they are infecting.

IV. Comparison of Eukaryotic & Prokaryotic Cells

The main difference between prokaryotes and eukaryotes is this: eukaryotic cells contain membrane-
bound organelles, such as the nucleus, while prokaryotic cells do not. Differences in cellular structure of
prokaryotes and eukaryotes include the presence of mitochondria and chloroplasts, the cell wall, and the
structure of chromosomal in eukaryotes, and the absence of those organelles in prokaryotes. DNA.
Prokaryotes were the only form of life on Earth for millions of years until more complicated eukaryotic cells
came into being through the process of evolution.

Eukaryotic Cell Prokaryotic Cell

Nucleus Present Absent
Number of chromosomes More than one One--but not true
chromosome: Plasmids
Cell Type Usually multicellular Usually unicellular (some
cyanobacteria may be
multicellular)
True Membrane bound Present Absent
Nucleus
Example Animals and Plants Bacteria and Archaea
Genetic Recombination Meiosis and fusion of gametes Partial, undirectional
transfers DNA
Lysosomes and Present Absent
peroxisomes
Microtubules Present Absent or rare
Endoplasmic reticulum Present Absent
Mitochondria Present Absent
Cytoskeleton Present May be absent
DNA wrapping on proteins Eukaryotes wrap their DNA Multiple proteins act together
 around proteins called to fold and condense
histones. prokaryotic DNA. Folded DNA
is then organized into a
variety of conformations that
are supercoiled and wound
around tetramers of the HU
protein.
Ribosomes larger smaller
Vesicles Present Present
Golgi apparatus Present Absent
Chloroplasts Present (in plants) Absent; chlorophyll scattered
in the cytoplasm
Flagella Microscopic in size; Submicroscopic in size,
membrane bound; usually composed of only one fiber
arranged as nine doublets
surrounding two singlets
Permeability of Nuclear Selective Not present
Membrane
Plasma membrane with Yes Usually no
steroid
Cell wall Only in plant cells and fungi Usually chemically complexed
(chemically simpler)
Vacuoles Present Present
Cell size 10-100um 1-10um

V. Microbial Classification of Infectious Disease

The Diseases caused by germs and which may infect any part of the body are called Infectious Diseases. They
can spread by any means where there is a germ. They are caused by pathogenic microorganisms such as bacteria,
virus, parasites and fungi. Germs can be spread by direct or indirect contact. Vaccination, maintenance of proper
hygiene and medicines help in the prevention of infection.

 Bacterial infectious diseases

 Viral infectious diseases
 Parasitic infectious diseases
 Fungal infectious diseases
 Mycobacterial diseases
 Air borne diseases
 Food borne diseases
 Water borne diseases
 Mosquito and tick borne diseases
 Blood borne infectious diseases
 Childhood infectious diseases or pediatric infectious diseases
 Geriatric infectious diseases
 Nosocomial infections or hospital acquired infections
 Sexually transmitted diseases
 Allergic infectious diseases
 Neuro infectious diseases
 Transplant infectious diseases
 Topical infectious diseases
 Inflammatory infectious diseases
 Opportunistic infections
 Infectious diseases in pregnancy
  Deadly infectious diseases
 Rare infectious diseases
 Communicable infectious diseases
 Common infectious diseases
 Zoonotic Bacterial Diseases
 Tetanus
 Typhoid fever
 Cholera
 C. gattii infection collapsed
 Infection Control

REFERENCES:

https://www.cliffsnotes.com/study-guides/biology/microbiology/introduction-to-microbiology/a-brief-history-of-
microbiology

https://www.studyread.com/classification-of-bacteria/

https://micro.magnet.fsu.edu/cells/bacteriacell.html

https://www.diffen.com/difference/Eukaryotic_Cell_vs_Prokaryotic_Cell

https://www.omicsonline.org/conferences-list/classification-of-infectious-diseases