DIVYA MANASVI RAJ MEHTA

Microbiology Assignment

Topic: Introduction to Pharmaceutical Microbiology

Definition:
Microbiology is the study of microorganisms, which are unicellular or cell-
cluster microscopic organisms. This includes eukaryotes such as fungi and prostates,
and prokaryotes. Viruses, though not strictly classed as living organisms, are also
studied. In short; microbiology refers to the study of life and organisms that are too small
to be seen with the naked eye.

Introduction:
Microbiology typically includes the study of the immune system,
or Immunology. Generally, immune systems interact with pathogenic microbes; these two
disciplines often intersect which is why many colleges offer a paired degree such as
"Microbiology and Immunology".
Microbiology is a broad term which includes virology, mycology, parasitological,
bacteriology and other branches.
 Microbiology is researched actively, and the field is advancing continually.
We have probably only studied about one percent of all of the microbe species on Earth.
Although microbes were directly observed over three hundred years ago, the field of
microbiology can be said to be in its infancy relative to older biological disciplines such
as zoology and botany.

Scope:
The field of microbiology can be generally divided into several sub disciplines:

Microbial physiology: The study of how the microbial cell functions biochemically.
Includes the study of microbial growth, microbial metabolism and microbial cell structure.

Microbial genetics: The study of how genes are organized and regulated in microbes in
relation to their cellular functions. Closely related to the field of molecular biology.

Cellular microbiology: A discipline bridging microbiology and cell biology.

Medical microbiology: The study of the pathogenic microbes and the role of microbes in
human illness. Includes the study of microbial pathogenesis and epidemiology and is
related to the study of disease pathology and immunology.

Veterinary microbiology: The study of the role in microbes in veterinary medicine or

animal taxonomy.

Environmental microbiology: The study of the function and diversity of microbes in their
natural environments. Includes the study of microbial ecology, microbial-
mediated nutrient cycling, geomicrobiology, microbial diversity and bioremediation.
Characterisation of key bacterial habitats such as
the rhizosphere and phyllosphere, soil and groundwater ecosystems, open oceans or
extreme environments (extremophiles).
Evolutionary microbiology: The study of the evolution of microbes. Includes the study of
bacterial systematics and taxonomy.

Industrial microbiology: The exploitation of microbes for use in industrial processes.

Examples include industrial fermentation and wastewater treatment. Closely linked to
thebiotechnology industry. This field also includes brewing, an important application of
microbiology.

Aeromicrobiology: The study of airborne microorganisms.

Food microbiology: The study of microorganisms causing food spoilage and food borne
illness. Using microorganisms to produce foods, for example by fermentation.

Pharmaceutical microbiology:The study of microorganisms causing pharmaceutical

contamination and spoil
History:
In 1546 Girolamo Fracastoro proposed that epidemic diseases were caused by
transferable seedlike entities that could transmit infection by direct or indirect contact or
even without contact over long distances.

Bacteria, and other microorganisms, were first observed by Antonie van

Leeuwenhoek in 1676 using a single-lens microscope of his own design. In doing
so Leeuwenhoek made one of the most important discoveries in biology and initiated the
scientific fields of bacteriology and microbiology. The name "bacterium" was introduced
much later, by Ehrenberg in 1828, derived from the Greek βακτηριον meaning "small
stick".

The field of bacteriology (later a subdiscipline of microbiology) is generally

considered to have been founded by Ferdinand Cohn (1828–1898), a botanist whose studies
on algae and photosynthetic bacteria led him to describe several bacteria
including Bacillus and Beggiatoa. Cohn was also the first to formulate a scheme for
the taxonomic classification of bacteria.

Microscopy:
Microscopy is the technical field of using microscopes to view samples or
objects. There are three well-known branches of microscopy, optical, electron and scanning
probe microscopy.

Optical and electron microscopy involve the diffraction, reflection,

or refraction of electromagnetic radiation/electron beam interacting with the subject of
study, and the subsequent collection of this scattered radiation in order to build up an
image. This process may be carried out by wide-field irradiation of the sample (for example
standard light microscopy and transmission electron microscopy) or by scanning of a fine
beam over the sample (for example confocal laser scanning microscopy and scanning
electron microscopy). Scanning probe microscopyinvolves the interaction of a scanning
probe with the surface or object of interest.

The development of microscopy revolutionized biology and remains an

essential tool in that science, along with many others including materials science and
numerous engineering disciplines.

Classification Of Microbes:

Microbes - bacteria, fungi, algae, protozoa and viruses -

affect every aspect of life on earth. They have an amazing diversity of form and can exist in
a wide range of habitats from hot springs to the icy wastes of Antarctica and inside the
bodies of animals and plants. Microbes cause diseases like 'flu or malaria, but most are
completely harmless.

They are essential to the cycling of nutrients in the ecosystems of the planet.
Microbial activity is exploited for the benefit of humankind in many ways, such as the
production of medicines, food and enzymes, in the clean-up of sewage and other wastes and
in the exciting advances resulting from developments in molecular biology techniques.

Until the middle of the 19th century all living organisms were classified into two
groups, animals and plants. There were problems with this simplistic system, for example
fungi look like plants but they do not photosynthesise. Over the years, a number of
classification systems were put forward; the most well known is the Five Kingdom system
which groups all living things by cell type, level of organization and nutrition as follows:

1. Animals
2. Plants
3. Fungi
4. Protoctista (algae and protozoa)
5. Monera (bacteria)

The first four groups represent eukaryotes (cells have a nucleus and membrane-bound
organelles). The Monera includes all prokaryotes (cells lack a nucleus and membrane-
bound organelles).

In the light of recent advances in molecular biology, which allow the comparison of the
sequencing of ribosomal RNA of organisms, a new classification system is preferred by
scientists. It is based on three lines of descent from a common ancestor. Each group is
called a Domain:

1.Bacteria (true bacteria) - prokaryotes

2. Archaea (archaebacteria) - prokaryotes
3. Eukarya – eukaryotes
The archaebacteria are prokaryotic in general structure and share many bacterial
characteristics. However they share with eukaryotes a some ribosomal sequences that are
not found in bacteria.
Viruses are not usually included in classification systems as they are non-cellular and they
are dependent on a host cell for their replication and metabolic processes.
Within their domains, identification of microbes begins with their physical appearance,
followed by biochemical and genetic tests.

Bacteria:

Bacteria consist of only one cell, but they're a very complex group of living
things. Some bacteria can live in temperatures above the boiling point and in cold below
the freezing point.

There are thousands of species of bacteria, but all of them are basically one of
three different shapes. Some are rod- or stick-shaped; others are shaped like little balls.
Others still are helical or spiral in shape. Some bacteria cells exist as individuals while
others cluster together to form pairs, chains, squares or other groupings.

Some bacteria can make their own food from sunlight, just like plants. Also
like plants, they give off oxygen. Other bacteria absorb food from the material they live on
or in. Some of these bacteria can live off iron or sulfur! The bacteria that live in your
stomach absorb nutrients from the digested food you've eaten.
Some bacteria move about their environment by means of long, whip-like structures called
flagella. They rotate their flagella like tiny outboard motors to propel themselves through
liquid environments. They may also reverse the direction in which their flagella rotate so
that they tumble about in one place. Other bacteria secrete a slime layer and ooze over
surfaces like slugs. Others stay almost in the same spot.
Bacteria live on or in just about every material and environment on Earth from soil to
water to air, and from your body to the Arctic ice to the Sahara deserts. Each square
centimeter of your skin averages about 100,000 bacteria. A single teaspoon of soil contains
more than a billion (1,000,000,000) bacteria.
Stucture of Bacterial Cell:
Viruses:
A virus is too small to be seen without a microscope. A virus is basically a tiny
bundle of genetic material carried in a shell called the viral coat. Some viruses have an
additional layer around this coat called an envelope. That's basically all there is to viruses.

There are thousands of different viruses that come in many shapes. Many are
multi-sided or polyhedral. If you've ever looked closely at a cut gem, like the diamond in an
engagement ring, you've seen an example of a polyhedral shape. Unlike the diamond in a
ring, however, a virus does not taper to a point, but is shaped the same all around. Other
viruses are shaped like spiky ovals or bricks with rounded corners. Some are like skinny
sticks while others look like pieces of looped string. Some are more complex and shaped
like little spaceship landing pods.

Viruses are found on or in just about every material and environment on Earth
from soil to water to air. They're basically found anywhere there are cells to infect. Viruses
can infect every living thing. However, viruses tend to be somewhat picky about what type
of cells they infect. Plant viruses are not equipped to infect animal cells, for example,
though a certain plant virus could infect a number of related plants. Sometimes, a virus
may infect one animal and do no harm, but cause a great deal of damage when it gets into a
different but closely related animal.
Viruses exist to reproduce only. To do that, they have to take over suitable host cells. Upon
landing on a suitable host cell, a virus gets its genes inside the cell either by tricking the
host cell to pull it inside, or by connecting its viral coat with the host cell wall or membrane
and releasing its genes inside, or by injecting their genes into the host cell's DNA.

The viral genes are then copied many times, using the process the host cell
would normally use to reproduce its own DNA. The new viral genes then come together
and assemble into whole new viruses. The new viruses are either released from the host cell
without destroying the cell or eventually build up to a large enough number that they burst
the host cell.

Stucture of a Virus:
Electron Microscopy:

Transmission electron microscope (TEM):

The original form of electron microscope,

the transmission electron microscope (TEM) uses a high voltage electron beam to create an
image. The electrons are emitted by an electron gun, commonly fitted with
a tungsten filament cathode as the electron source. The electron beam is accelerated by
an anode typically at +100 keV (40 to 400 keV) with respect to the cathode, focused
by electrostatic and electromagnetic lenses, and transmitted through the specimen that is in
part transparent to electrons and in part scatters them out of the beam. When it emerges
from the specimen, the electron beam carries information about the structure of the
specimen that is magnified by the objective lens system of the microscope. The spatial
variation in this information (the "image") is viewed by projecting the magnified electron
image onto a fluorescent viewing screen coated with a phosphor or scintillator material
such as zinc sulfide. The image can be photographically recorded by exposing
a photographic film or plate directly to the electron beam, or a high-resolution phosphor
may be coupled by means of a lens optical system or a fibre optic light-guide to the sensor
of a CCD (charge-coupled device) camera. The image detected by the CCD may be
displayed on a monitor or computer.

Resolution of the TEM is limited primarily by spherical aberration, but a new

generation of aberration correctors have been able to partially overcome spherical
aberration to increase resolution. Hardware correction of spherical aberration for the High
Resolution TEM (HRTEM) has allowed the production of images with resolution below
0.5 Ångström (50 picometres at magnifications above 50 million times. The ability to
determine the positions of atoms within materials has made the HRTEM an important tool
for nano-technologies research and development.

Scanning electron microscope (SEM):

 Unlike the TEM, where electrons of the high
voltage beam carry the image of the specimen, the electron beam of the Scanning Electron
Microscope(SEM)[9] does not at any time carry a complete image of the specimen. The
SEM produces images by probing the specimen with a focused electron beam that is
scanned across a rectangular area of the specimen (raster scanning). At each point on the
specimen the incident electron beam loses some energy, and that lost energy is converted
into other forms, such as heat, emission of low-energy secondary electrons, light emission
(cathodoluminescence) or x-ray emission. The display of the SEM maps the varying
intensity of any of these signals into the image in a position corresponding to the position of
the beam on the specimen when the signal was generated. In the SEM image of an ant
shown at right, the image was constructed from signals produced by a secondary electron
detector, the normal or conventional imaging mode in most SEMs.
Generally, the image resolution of an SEM is about an order of magnitude poorer than that
of a TEM. However, because the SEM image relies on surface processes rather than
transmission, it is able to image bulk samples up to many centimetres in size and
(depending on instrument design and settings) has a great depth of field, and so can
produce images that are good representations of the three-dimensional shape of the sample.

Reflection electron microscope (REM):

In the Reflection Electron Microscope (REM) as

in the TEM, an electron beam is incident on a surface, but instead of using the transmission
(TEM) or secondary electrons (SEM), the reflected beam of elastically scattered electrons is
detected. This technique is typically coupled with Reflection High Energy Electron
Diffraction (RHEED) and Reflection high-energy loss spectrum (RHELS). Another
variation is Spin-Polarized Low-Energy Electron Microscopy (SPLEEM), which is used for
looking at the microstructure of magnetic domains.

Scanning transmission electron microscope (STEM):

The STEM rasters a focused

incident probe across a specimen that (as with the TEM) has been thinned to facilitate
detection of electrons scattered through the specimen. The high resolution of the TEM is
thus possible in STEM. The focusing action (and aberrations) occur before the electrons hit
the specimen in the STEM, but afterward in the TEM. The STEMs use of SEM-like beam
rastering simplifies annular dark-field imaging, and other analytical techniques, but also
means that image data is acquired in serial rather than in parallel fashion.

Low voltage electron microscope (LVEM):

The low voltage electron microscope (LVEM) is a

combination of SEM, TEM and STEM in one instrument, which operated at relatively low
electron accelerating voltage of 5 kV. Low voltage increases image contrast which is
especially important for biological specimens. This increase in contrast significantly
reduces, or even eliminates the need to stain. Sectioned samples generally need to be
thinner than they would be for conventional TEM (20-65nm). Resolutions of a few nm are
possible in TEM, SEM and STEM modes.

Dark Field Microscopy:

Dark field microscopy (dark ground microscopy) describes
microscopy methods, in both light and electron microscopy, which exclude the unscattered
beam from the image. As a result, the field around the specimen (i.e. where there is no
specimen to scatter the beam) is generally dark.

Dark field microscopy is a very simple yet effective

technique and well suited for uses involving live and unstained biological samples, such as a
smear from a tissue culture or individual water-borne single-celled organisms. Considering
the simplicity of the setup, the quality of images obtained from this technique is impressive.

The main limitation of dark field microscopy is the low light

levels seen in the final image. This means the sample must be very strongly illuminated,
which can cause damage to the sample.
Dark field microscopy techniques are almost entirely free of artifacts, due to the nature of
the process. However the interpretation of dark field images must be done with great care
as common dark features of bright field microscopy images may be invisible, and vice
versa.

While the dark field image may first appear to be a

negative of the bright field image, different effects are visible in each. In bright field
microscopy, features are visible where either a shadow is cast on the surface by the incident
light, or a part of the surface is less reflective, possibly by the presence of pits or scratches.
Raised features that are too smooth to cast shadows will not appear in bright field images,
but the light that reflects off the sides of the feature will be visible in the dark field images.

Applications:

Conventional darkfield imaging:

Briefly, conventional darkfield imaging involves tilting

the incident illumination until a diffracted, rather than the incident, beam passes through a
small objective aperture in the objective lens back focal plane. Darkfield images, under
these conditions, allow one to map the diffracted intensity coming from a single collection
of diffracting planes as a function of projected position on the specimen, and as a function
of specimen tilt.
In single crystal specimens, single-reflection darkfield images of a specimen tilted just off
the Bragg condition allow one to "light up" only those lattice defects, like dislocations or
precipitates, which bend a single set of lattice planes in their neighborhood. Analysis of
intensities in such images may then be used to estimate the amount of that bending. In
polycrystalline specimens, on the other hand, darkfield images serve to light up only that
subset of crystals which is Bragg reflecting at a given orientation.

Weak beam imaging:

Weak beam imaging involves optics similar to conventional

darkfield, but use of a diffracted beam harmonic rather than the diffracted beam itself.
Much higher resolution of strained regions around defects can be obtained in this way.

Low and high angle annular darkfield imaging:

Annular darkfield imaging requires one

to form images with electrons diffracted into an annular aperture centered on, but not
including, the unscattered beam. For large scattering angles in a scanning transmission
electron microscope, this is sometimes called Z-contrast imaging because of the enhanced
scattering from high atomic number atoms.

Phase Contrast Microscopy:

Phase contrast microscopy is an optical

microscopy illumination technique in which small phase shifts in the light passing through
a transparent specimen are converted into amplitude or contrast changes in the image.

A phase contrast microscope does not require staining to view the slide. This type
of microscope made it possible to study the cell cycle.
As light travels through a medium other than vacuum, interaction with this medium causes
its amplitude and phase to change in a way which depends on properties of the medium.
Changes in amplitude give rise to familiar absorption of light which gives rise to colours
when it is wavelength dependent. The human eye measures only the energy of light arriving
on the retina, so changes in phase are not easily observed, yet often these changes in phase
carry a large amount of information.
The same holds in a typical microscope, i.e., although the phase variations introduced by
the sample are preserved by the instrument (at least in the limit of the perfect imaging
instrument) this information is lost in the process which measures the light.
 In order to make phase variations observable, it is necessary to combine the light
passing through the sample with a reference so that the resulting interference reveals the
phase structure of the sample.

Common Communicable Diseases:

Sr.No. Name of Mode of Incubation Symptoms Management

Disease Spread period of the Patient
1. Cholera Food and 1-5 days (i) Sudden Dehydration
Water onset of can be
severe, dangerous,
watery so give plenty
diarrhoea. of fluids. Give
The faeces Oral
look like rice rehydration
water solution
(ii) Vomitting (ORS).
(iii) Cramps
in the legs
(iv) Patient
feels very
thirsty
2. Typhoid Food and 14-21 days (i) Severe Blood culture
Water headache and other
(ii) Fever tests
with low should be
pulse done. Give the
(iii) Dry white prescribed
coated tongue medication to
the patient.
3. Hepatitis Food and 20-35 days (i) Fever Give a
(jaundice) Water (ii) Dark carbohydrates
yellow urine rich diet.
(iii) Yellowish Keep the
tinge in eyes patient in bed
(iv) General as long
paleness as there is
fever and till
appetite
returns to
normal.
4. Influenza Air 1-3 days (i) Fever Control the
(Flu) (ii) Cold, fever with
cough, medicines
sneezing and cough
(iii) Headache with steam
and body inhalation.
ache
(iv) Nausea
5. Tuberculosis Air 4-6 days (i) Persistent Treatment is
(T.B) cough prolonged so
 (ii) Loss of constant
weight and monitoring by
appetite the doctor
(iii) Excessive is essential.
weakness
(iv) Rapid
pulse
(v) Chest pain
(vi) Breath
has bad smell
6. Malaria Mosquito 10-14 days (i) Fever Get the blood
bite (ii) test done to
Alternating confirm
chill and malaria. Then
perspiration give
(iii) Headache prescribed
and bodyache medicines
(iv) Nausea
(v) Vomiting
7. Tetanus Wound 4 days to 2 (i) Put a ball of
exposed to weeks Restlessness cotton
dust or (ii) Headache between teeth
rusted item (iii) Fever0 to prevent
(iv) Stff neck biting of
(v) Difficulty tongue.
in chewing
and
swallowing
(vi) Spasm of
muscles of
jaw and face
(vii) Bending
of back in
shape of bow
(viii)Severe
pain

Bacterial Resistance:

Our body’s immune system uses specially designed cells to

locate and shut down microscopic invaders like bacteria, usually stopping them before they
can cause trouble. We get sick — what is called a bacterial “infection” — when bacteria in
our body reproduce faster than our immune system can kill them.

Antibiotics are powerful bacteria-killing drugs that help our bodies regain the upper hand
when a bacterial infection develops. Today, there are hundreds of antibiotics in use, most
tailored to treat a specific kind of bacterial infection. (That’s why taking unused antibiotics
prescribed for one kind of bacterial infection won’t necessarily work against another.
Never “save” anti-biotics; always finish the full course of treatment as prescribed.)
Doctors have noticed that some bacteria are getting tougher to kill. The usual antibiotic
drugs don’t seem to work as well — or work at all. Such bacteria are said to be resistant.
Bacterial resistance makes an infection much harder to treat. Higher doses or stronger
drugs may be required. In extreme cases, bacterial resistance can be fatal.
xperts like the scientists at the Centers for Disease Control and Prevention (CDC) agree
that the overprescription and misuse of anti-biotic drugs are the main causes of bacterial
resistance. The CDC says that up to half of the roughly 100 million prescriptions for
antibiotics written each year are unnecessary.

Other causes for Bacterial Resistance:

• Bacteria reproduce by dividing to create copies of themselves; sometimes the copies

aren’t exact and the new organism has different characteristics than the original.
These characteristics could make the new organism resistant to an antibiotic.
• Bacteria have the ability to “share” resistant characteristics with each other outside
of reproduction. This makes it possible to transfer resistance from one person to
another through exposure to resistant bacteria.
• Bacterial resistance in humans may be increased by the use of preventive antibiotics
in animal feed. In 1995, an estimated 4.5 million pounds of antibiotics were used to
reduce the spread of disease and enhance the growth of cattle, swine and poultry.
The U.S. Food and Drug Administration (FDA) is now reviewing this practice to
determine its potential health impact.
• The U.S. Food and Drug Administration (FDA) convened a panel of experts to
examine the possible role of antibacterial hand and body wash products in
promoting bacterial resistance. The panel reviewed the available science and
determined that antibacterial wash products were not a public health concern.

Tips for fighting bacterial resistance:

1. Never take an antibiotic for viral infections such as colds or flu.

Don’t ask your doctor to prescribe antibiotics if he or she doesn’t think they are
necessary.

2. If an antibiotic is called for, use it exactly as the doctor prescribes.

Follow the doctor’s treatment instructions and finish the full amount of antibiotic
prescribed. Don’t stop taking the medication just because you are feeling better. Never save
anti-biotics to treat yourself or others later.

3. Always wash your hands thoroughly.

Scrub your hands vigorously for 10 to 15 seconds using soap and warm water. Many
leading brands have the word “antibacterial” on the label. (Remember to wash between
your fingers, where germs accumulate.)

4. Always handle food correctly.

Basic sanitation and proper food handling can go a long way toward preventing foodborne
illness.
Stay safe:
Keep your hands, utensils and food preparation surfaces clean.
Avoid cross-contamination; don’t let raw meat, poultry and fish — or their juices — come
into contact with other foods.
Cook foods to the proper temperature to kill off dangerous microorganisms.
Refrigerate foods promptly to keep harmful bacteria from growing and multiplying.

5. Get vaccinated.
If you’re 65 or older or you have a chronic illness, you should get vaccinated for
pneumococcal pneumonia. It’s a major cause of death in older adults.

Bibliography
www.wikipedia.org/microbiology

www.wikipedia.org/microscopy

www.textbookof bacteriology.net

www.nclnet.org/microbes