Assignment no.

1
Subject: microbiology
Topic: bacteria
Question no.1
Cellular morphology of bacteria:
(a) structure external to cell wall
(b) cell wall
(c) structure internal to cell wall
Answer:
Introduction
 Bacteria are unicellular prokaryotes.
 They lack true nucleus and other membrane bound
organelles.
 Some species form colonies.
 Most prokaryotes have diameter between 1-5 µm.
MORPHOLOGY
Bacteria are amazingly heterogeneous
collection of microscopic organisms, occurring in all possible
shapes ranging from spheres, rods, spirals to pleomorphic mass.
They are divided into the following groups;
COCCUS

These bacteria are round but can also be oval, elongated or

flattened on one side. They exist either single round
cell(Micrococcus)

In pairs (diplococcus) in chains (Streptococcus),in grape

like clusters (Staphylococcus), in cubical masses of 8 or more
cells(Sarcina). These arrangements result when the daughter
cells formed after division by binary fission in both the planes do
not separate from each other.

BACILLUS

bacteria with rod shaped cells ranging in size from

about 0.2-1.5 μm in diameter and 1-10 μm in length. They also
occur either singly, in pairs (Diplobacillus), in chains
(Streptobacillus), or in palisade arrangement. Mycobacterium, E.
coli, Salmonella, and Clostridium are some of the common
examples of bacilli. The rods either have smooth rounded ends
(E. coli) or tapered(pointed) ends (Fusobacterium), which can be
rigid or flexible. Still other kind of bacteria appear oval and look
more like cocci(Coccobacilli).

SPIRAL BACTERIA

they have one or more twists and are never

straight. When they have a single turn (coma shaped) they are
called Vibrios. Spirilla have a helical shape and a rigid body and
appear coiled like a cork-screw with 1-5 complete turns. They
range from 10-50 μm in length. e.g. Vibrio Cholerae, and Spirillum
undulum.

STALKED BACTERIA

These are unicellular bacteria having well defined

stalks. Sometimes, stalk is a protrusion of the cell as in
Caulobacter, while in others it forms as a result of secretion from
the cell(Gallionella). Typically, these bacteria have a knob-like
base, which is sticky, and the cells join each other at the base
forming a rosette-like structure.
BUDDING BACTERIA
 These bacteria have a thin and small tubular
structure, which elongates and swells forming a new cell. This
type of division results in a network of globular cells, e.g.
Rhodomicrobium.

(a) STRUCTURE EXTERNAL TO CELL WALL

Following are the structure external to cell wall:

GLYCOCALYX
Many prokaryotes secrete on their surface a substance
called glycocalyx. Glycocalyx (meaning sugar coat) is a
general term that use for the substance that surrounds cells.
It is a
 Viscous (sticky)
 Gelatinous polymer (external to cell that is
composed of polysaccharides or polypeptide or
both)
If substance is organized and is firmly attached to the cell
wall, the glycocalyx is describe as a capsule. The presence of
capsule can be determined by using negative staining. And if the
substance is unorganized and only loosely attached to the cell
wall, the glycocalyx is describe as a slime layer.
FUNCTION OF CAPSULE
 Capsule often protect pathogenic bacteria from
phagocytosis by the cells of the host.
 It prevents phagocytosis and allows the bacterium to
adhere to and colonize the respiratory tract.
 It makes bacteria resistant to antibiotics.
 It can also protect a cell against dehydration.
 Its viscosity may inhibit the movement of nutrients out of
the cell.
EXTRACELLULAR POLYSACCHARIDES (EPS)
A glycocalyx made up of sugars is called as extracellular
polysaccharides. The ESP enables a bacterium to survive by
attaching to various surfaces in its natural environment in order to
survive. Through attachment, bacteria can grow on diverse
surfaces such as rocks.
FLAGELLA
Some prokaryotic cells have flagella (flagellum,
meaning whip), which are long filamentous appendages that
propel bacteria. Bacteria lack flagella are referred to as
atrichous. Those that have flagella may have one of four
arrangements of flagella.
 Monotrichous (a single polar flagellum)
 Amphitrichous (a tuft of flagella at each end of the
cells)
 Lophotrichous (two or more flagella at one or both
ends of the cells)
 Peritrichous (flagella distributed over the entire cell)
There are three basic parts of the flagellum, the long outermost
region of filament, is constant in diameter and contains the
globular (roughly spherical) protein flagellin arranged in several
chains that intertwine and form a helix around a hollow core. In
most bacteria, filaments are not covered by a membrane or
sheath, as in eukaryotic cells. The filament is attached to a slightly
wider hook, consisting of a different protein. The third portion of a
flagellum is the basal body, which anchors the flagellum to the cell
wall and plasma membrane. The basal body is composed of a
small central rod inserted into a series of rings.
Gram negative bacteria contains two pairs of rings; the outer
pair of rings is anchored to various portions of the cells wall, and
the inner pair of rings is anchored to the plasma membrane. Gram
positive bacteria, only the inner pair is present.
FUNCTIONS
 The primary function of flagella is locomotion. Flagella
are used for swimming through water, bacterial gliding
and twitching. These movements also change the
buoyancy to allow vertical motion. Swimming bacteria
frequently move near 10 body lengths per second and
a few as fast as fish as 100. This makes them at least
as fast as fish, on a relative scale. In twitching motility,
bacteria use their pili as a grappling hook. Bacteria
repeatedly extend it, anchor it and then retract it with
remarkable force.
 Bacteria can detect a chemical signal with the help of
flagella and move in its response. Such type of
behavior called chemotaxis.

AXIAL FILAMENTS
 Spirochetes are the group of bacteria that
have unique structure and motility. Spirochete move by means of
axial filaments, or endoflagella, bundles of fibrils that arise at the
ends of cells beneath an outer sheath and spiral around the cell.
Axial filaments which are anchored at one end of the spirochete,
have a structure similar to that of flagella. The rotation of the
filaments produces a movement of the outer sheath that propels
the spirochete in a spiral motion.
FIMBRIAE AND PILI
Many gram negative bacteria contain hair like
appendages that are shorter, straighter and thinner than flagella
and are used for attachment and transfer of DNA rather than for
motility. These structure which consists of proteins called pilin are
arranged helically around a central core, are divided into two
types, fimbriae and pili, having very different functions.
Fimbriae can occur at the poles of the bacterial cell, or they
evenly distributed over the entire surface of the cell. They can
vary from a few to hundred per cell. like the glycocalyx, fimbriae
enable a cell to adhere to surfaces, including the surface of the
surface of the other cell.
Pili (singular pilus) are usually longer than fimbriae and number
one or two per cell. Pili join bacterial cells in preparation for the
transfer of DNA from one cell to another, a process called as
conjugation. For this reason, they are sometimes called
conjugation pili.
(a) THE CELL WALL
 The cell wall of the bacterial cell is a complex, Simi rigid
structure responsible for the shape of the cell.
 The cell surrounds the underlying, fragile plasma membrane
and protects it and the interior of the cell from adverse
 changes in the outside environment. Almost all prokaryotes
have cell walls.
 The major function of the cell wall is to prevent bacterial cells
from rupturing when the water pressure inside the cell is
greater than outside the cell.
 It also helps to maintain the shape of a bacterium and serve
as point of anchoring for flagella.
 Clinically cell is important because it contributes to the ability
of some species to cause diseases and is the site of action
of some antibiotics.
COMPOSITION AND CHARACTERISTICS
The bacterial cell wall is composed of a macromolecular
network called peptidoglycan (also known as murein), which is
present either alone or in combination with other substances.
Peptidoglycan consists of a repeating disaccharides attached
by polypeptides to form a lattice that surrounds and protect the
entire cell. The disaccharide portion is made up of
monosaccharides called N-acetyl glucosamine (NAG) and N-
acetylmuramic acid (NAM) (from murus, meaning wall), which
are related to glucose. Peptidoglycan is responsible for the
rigidity of the bacterial cell wall and for the determination of cell
shape. It is relatively porous and is not considered to be a
permeability barrier for small substrates. While all bacterial cell
walls (with a few exceptions e.g. extracellular parasites such as
Mycoplasma) contain peptidoglycan, not all cell walls have the
same overall structures. Since the cell wall is required for
bacterial survival, but is absent in some eukaryotes, several
antibiotics (notably the penicillin and cephalosporins) stop
bacterial infections by interfering with cell wall synthesis, while
having no effects on human cells which have no cell wall, only
a cell membrane. There are two main types of bacterial cell
 walls, those of gram-positive bacteria and those of gram-
negative bacteria, which are differentiated by their Gram
staining characteristics. For both these types of bacteria,
particles of approximately 2 nm can pass through the
peptidoglycan. If the bacterial cell wall is entirely removed, it is
called a protoplast while if it's partially removed, it is called a
spheroplast.
β-Lactam antibiotics such as penicillin inhibit the formation of
peptidoglycan cross-links in the bacterial cell wall. The enzyme
lysozyme, found in human tears, also digests the cell wall of
bacteria and is the body's main defense against eye infections.
The gram-positive cell wall
Gram-positive cell walls are thick and the peptidoglycan (also
known as murein) layer constitutes almost 95% of the cell wall in
some gram-positive bacteria and as little as 5-10% of the cell wall
in gram-negative bacteria. The gram-positive bacteria take up the
crystal violet dye and are stained purple. The cell wall of some
gram-positive bacteria can be completely dissolved by lysozymes
which attacks the bonds between N-acetylmuramic acid and N-
acetyl glucosamine. In other gram-positive bacteria, such as
Staphylococcus aureus, the walls are resistant to the action of
lysozymes. They have O-acetyl groups on carbon-6 of some
muramic acid residues. The matrix substances in the walls of
gram-positive bacteria may be polysaccharides or teichoic acids.
The latter are very widespread, but have been found only in gram-
positive bacteria.
There are two main types of teichoic acid: ribitol teichoic acids
and glycerol teichoic acids. These acids are polymers of ribitol
phosphate and glycerol phosphate, respectively, and only located
on the surface of many gram-positive bacteria. However, the
exact function of teichoic acid is debated and not fully understood.
A major component of the gram-positive cell wall is lipoteichoic
acid. One of its purposes is providing an antigenic function. The
lipid element is to be found in the membrane where its adhesive
properties assist in its anchoring to the membrane.
The gram-negative cell wall
Gram-negative cell walls are much thinner than the gram-positive
cell walls, and they contain a second plasma membrane
superficial to their thin peptidoglycan layer, in turn adjacent to the
cytoplasmic membrane. Gram-negative bacteria are stained as
pink color. The chemical structure of the outer membrane's
lipopolysaccharide is often unique to specific bacterial subspecies
and is responsible for many of the antigenic properties of these
strains.

ATYPICAL CELL WALLS

Among prokaryotes contain types of
cells have no wall or have very little wall material. these include
member of genus Mycoplasma and related organism.
Mycoplasmas are the smallest known bacteria that can grow and
reproduce outside living host cells. Because their size and
because they have no cell walls, they pass through never most
bacterial filters and were first mistaken for viruses. There plasma
membranes are unique among bacteria in having lipids called
sterol, which are thought to help to protect from lysis(rupture).
Archae may lack walls or may have usual walls composed of
polysaccharides and proteins but not peptidoglycans.
STRUCTURE INTERNAL TO CELL WALL
Plasma membrane
 The plasma membrane or bacterial cytoplasmic membrane
is composed of a phospholipid bilayer and thus has all of the
general functions of a cell membrane such as acting as a
permeability barrier for most molecules and serving as the
location for the transport of molecules into the cell.
 In addition to these functions, prokaryotic membranes also
function in energy conservation as the location about which
a proton motive force is generated.
 Unlike eukaryotes, bacterial membranes (e.g. Mycoplasma
and methanotrophs) generally do not contain sterols.
However, many microbes do contain structurally related
compounds called hopanoids which likely fulfill the same
function. bacteria can have a wide variety of fatty acids
within their membranes. Along with typical saturated and
unsaturated fatty acids, bacteria can contain fatty acids with
additional methyl, hydroxy or even cyclic groups. The
relative proportions of these fatty acids can be modulated by
the bacterium to maintain the optimum fluidity of the
membrane (e.g. following temperature change).
 As a phospholipid bilayer, the lipid portion of the outer
membrane is impermeable to charged molecules. However,
channels called porins are present in the outer membrane
that allow for passive transport of many ions, sugars and
amino acids across the outer membrane. These molecules
are therefore present in the periplasm, the region between
the cytoplasmic and outer membranes. The periplasm
 contains the peptidoglycan layer and many proteins
responsible for substrate binding or hydrolysis and reception
of extracellular signals. The periplasm is thought to exist in a
gel-like state rather than a liquid due to the high
concentration of proteins and peptidoglycan found within it.
Ribosomes and other multiprotein complexes
In most bacteria the most numerous intracellular structure is the
ribosome, the site of protein synthesis in all living organisms. All
prokaryotes have 70S (where S=Svedberg units) ribosomes while
eukaryotes contain larger 80S ribosomes in their cytosol. The 70S
ribosome is made up of a 50S and 30S subunits. The 50S subunit
contains the 23S and 5S rRNA while the 30S subunit contains the
16S rRNA. These rRNA molecules differ in size in eukaryotes and
are complexed with a large number of ribosomal proteins, the
number and type of which can vary slightly between organisms.
While the ribosome is the most commonly observed intracellular
multiprotein complex in bacteria other large complexes do occur
and can sometimes be seen using microscopy.
Cytoskeleton
The prokaryotic cytoskeleton is the collective name for all
structural filaments in prokaryotes. It was once thought that
prokaryotic cells did not possess cytoskeletons, but recent
advances in visualization technology and structure determination
have shown that filaments indeed exist in these cells. In fact,
homologues for all major cytoskeletal proteins in eukaryotes have
been found in prokaryotes. Cytoskeletal elements play essential
roles in cell division, protection, shape determination, and polarity
determination in various prokaryotes.
Inclusions
Inclusions are considered to be nonliving components of the cell
that do not possess metabolic activity and are not bounded by
membranes. The most common inclusions are glycogen, lipid
droplets, crystals, and pigments. Volutin granules are cytoplasmic
inclusions of complexed inorganic polyphosphate. These granules
are called metachromatic granules due to their displaying the
metachromatic effect; they appear red or blue when stained with
the blue dyes methylene blue or toluidine blue.
Gas vacuoles
Gas vacuoles are membrane-bound, spindle-shaped vesicles,
found in some planktonic bacteria and Cyanobacteria, that
provides buoyancy to these cells by decreasing their overall cell
density. Positive buoyancy is needed to keep the cells in the
upper reaches of the water column, so that they can continue to
perform photosynthesis. They are made up of a shell of protein
that has a highly hydrophobic inner surface, making it
impermeable to water (and stopping water vapour from
condensing inside) but permeable to most gases. Because the
gas vesicle is a hollow cylinder, it is liable to collapse when the
surrounding pressure increases. Natural selection has fine-tuned
the structure of the gas vesicle to maximize its resistance to
buckling, including an external strengthening protein, GvpC,
rather like the green thread in a braided hosepipe.
Microcompartments
Bacterial microcompartments are
widespread, membrane-bound organelles that are made of a
protein shell that surrounds and encloses various enzymes.
provide a further level of organization; they are compartments
within bacteria that are surrounded by polyhedral protein shells,
rather than by lipid membranes. These "polyhedral organelles"
localize and compartmentalize bacterial metabolism, a function
performed by the membrane-bound organelles in eukaryotes.
Carboxysomes
Carboxysomes are bacterial microcompartments found in many
autotrophic bacteria such as Cyanobacteria, Knallgasbacteria,
Nitroso- and Nitrobacteria. They are proteinaceous structures
resembling phage heads in their morphology and contain the
enzymes of carbon dioxide fixation in these organisms (especially
ribulose bisphosphate carboxylase/oxygenase, RuBisCO, and
carbonic anhydrase).
Magnetosomes
Magnetosomes are bacterial microcompartments found in
magnetotactic bacteria that allow them to sense and align
themselves along a magnetic field (magnetotaxis).
Magnetosomes are composed of the mineral magnetite or greigite
and are surrounded by a lipid bilayer membrane. The morphology
of magnetosomes is species-specific. [citation needed]
Endospores
Perhaps the best known bacterial adaptation to stress is the
formation of endospores. Endospores are bacterial survival
structures that are highly resistant to many different types of
chemical and environmental stresses and therefore enable the
survival of bacteria in environments that would be lethal for these
cells in their normal vegetative form. It has been proposed that
endospore formation has allowed for the survival of some bacteria
for hundreds of millions of years (e.g. in salt crystals). Endospore
formation is limited to several genera of gram-positive bacteria
such as Bacillus and Clostridium. It differs from reproductive
spores in that only one spore is formed per cell resulting in no net
gain in cell number upon endospore germination. The location of
an endospore within a cell is species-specific and can be used to
determine the identity of a bacterium. Dipicolinic acid is a
chemical compound which composes 5% to 15% of the dry
weight of bacterial spores and is implicated in being responsible
for the heat resistance of endospores.
QUESTION NO. 2
BACTERIAL GROWTH CURVE
Bacteria are prokaryotic organisms that most commonly replicate
by the asexual process of binary fission.
There are four distinct phases of the growth curve: lag,
exponential (log), stationary, and death.
 The initial phase is the lag phase where bacteria are
metabolically active but not dividing.
 The exponential or log phase is a time of exponential growth.
 In the stationary phase, growth reaches a plateau as the
number of dying cells equals the number of dividing cells.
 The death phase is characterized by an exponential
decrease in the number of living cells.
DESCRIPTION
Lag Phase: This initial phase is characterized by cellular activity
but not growth. A small group of cells are placed in a nutrient rich
medium that allows them to synthesize proteins and other
molecules necessary for replication. These cells increase in size,
but no cell division occurs in the phase.
Exponential (Log) Phase: After the lag phase, bacterial cells
enter the exponential or log phase. This is the time when the cells
are dividing by binary fission and doubling in numbers after each
generation time. Metabolic activity is high as DNA, RNA, cell wall
components, and other substances necessary for growth are
generated for division. It is in this growth phase that antibiotics
and disinfectants are most effective as these substances typically
target bacteria cell walls or the protein synthesis processes of
DNA transcription and RNA translation.
Stationary Phase: Eventually, the population growth experienced
in the log phase begins to decline as the available nutrients
become depleted and waste products start to accumulate.
Bacterial cell growth reaches a plateau, or stationary phase,
where the number of dividing cells equal the number of dying
cells. This results in no overall population growth. Under the less
favorable conditions, competition for nutrients increases and the
cells become less metabolically active. Spore forming bacteria
produce endospores in this phase and pathogenic bacteria begin
to generate substances (virulence factors) that help them survive
harsh conditions and consequently cause disease.
Death Phase:
As nutrients become less available and waste products increase,
the number of dying cells continues to rise. In the death phase,
the number of living cells decreases exponentially and population
growth experiences a sharp decline. As dying cells lyse or break
open, they spill their contents into the environment making these
nutrients available to other bacteria. This helps spore producing
bacteria to survive long enough for spore production. Spores are
able to survive the harsh conditions of the death phase and
become growing bacteria when placed in an environment that
supports life.
QUESTION NO. 3
PHYSICAL AND CHEMICAL GROWTH REQUIREMENTS OF
BACTERIA
Following are the conditions require for the bacterial growth;
Bacterial Growth and Oxygen
 Bacteria, like all living organisms, require an environment
that is suitable for growth. This environment must meet
several different factors that support bacterial growth. Such
factors include oxygen, pH, temperature, and light
requirements. Each of these factors may be different for
different bacteria and limit the types of microbes that
populate a particular environment.
 Bacteria can be categorized based on their oxygen
requirement or tolerance levels. Bacteria that cannot survive
without oxygen are known as obligate aerobes. These
microbes are dependent upon oxygen, as they convert
oxygen to energy during cellular respiration. Unlike bacteria
that require oxygen, other bacteria cannot live in its
presence. These microbes are called obligate anaerobes
and their metabolic processes for energy production are
halted in the presence of oxygen.
 Other bacteria are facultative anaerobes and can grow with
or without oxygen. In the absence of oxygen, they utilize
either fermentation or anaerobic respiration for energy
production. Aero tolerant anerobes utilize anaerobic
respiration but are not harmed in the presence of oxygen.
Microaerophilic bacteria require oxygen but only grow where
oxygen concentration levels are low. Campylobacter jejuni is
an example of a microaerophilic bacterium that lives in the
digestive tract of animals and is a major cause of foodborne
illness in humans.
Bacterial Growth and pH
Another important factor for bacterial growth is pH. Acidic
environments have pH values that are less than 7, neutral
environments have values at or near 7, and basic environments
have pH values greater than 7. Bacteria that are acidophiles
thrive in areas where the pH is less than 5, with an optimal growth
value close to a pH of 3. These microbes can be found in
locations such as hot springs and in the human body in acidic
areas such as the vagina.
The majority of bacteria are neutrophiles and grow best in sites
with pH values close to 7. Helicobacter pylori is an example of a
neutrophile that lives in the acidic environment of the stomach.
This bacterium survives by secreting an enzyme that neutralizes
stomach acid in the surrounding area. Alkaliphiles grow optimally
at pH ranges between 8 and 10. These microbes thrive in basic
environments such as alkaline soils and lakes.
Bacterial Growth and Temperature
Temperature is another important factor for bacterial growth.
Bacteria that grow best in cooler environments are called
psycrophiles. These microbes prefer temperatures ranging
between 4°C and 25°C (39°F and 77°F). Extreme psycrophiles
thrive in temperatures below 0°C/32°F and can be found in places
such as arctic lakes and deep ocean waters.
Bacteria that thrive in moderate temperatures (20-45°C/68-113°F)
are called Mesophiles. These include bacteria that are part of the
human microbiome which experience optimum growth at or near
body temperature (37°C/98.6°F).
Thermophiles grow best in hot temperatures (50-80°C/122-176°F)
and can be found in hot springs and geothermal soils. Bacteria
that favor extremely hot temperatures (80°C-110°C/122-230°F)
are called hyperthermophiles.
Bacterial Growth and Light
Some bacteria require light for growth. These microbes have light-
capturing pigments that are able to gather light energy at certain
wavelengths and convert it to chemical energy. Cyanobacteria are
examples of photoautotrophs that require light for photosynthesis.
These microbes contain the pigment chlorophyll for light
absorption and oxygen production through photosynthesis.
Cyanobacteria live in both land and aquatic environments and can
also exist as phytoplankton living in symbiotic relationships with
fungi (lichen), protists, and plants. Other bacteria, such as purple
and green bacteria, do not produce oxygen and utilize sulfide or
sulfur for photosynthesis. These bacteria contain
bacteriochlorophyll, a pigment capable of absorbing shorter
wavelengths of light than chlorophyll.
QUESTION NO. 4
MATERIAL AND EQUIPMENT USE IN MICROBIOLOGY
AUTOCLAVE;
Instrument Uses autoclave used for sterilization of glass ware and
media auto-destruct syringes specimen collection
Bijou bottle
a cylindrical small glass bottle with a screw cap used as a culture
medium holder Biosafety cabinet used to work with dangerous
organisms and to work sterile Blood collection bottle to collect
blood by venipuncture Brittany a process of free from spore-
bearing bacteria Bunsen burner used to work aseptic on the
bench
Candle jar
historically used for anaerobiosis; a lit candle was placed in as
airtight jar such that when it went out it would be because it used
up all the available oxygen
Castaneda's medium / Castaneda's bottle
used for simultaneous solid and liquid cultures in many bottles
Durham's tube
used to detect gas production in sugar fermentation media; the
tube is placed in an inverted fashion so that gases produced get
trapped in it and do not float away to the surface
Gas-pak
releases gases to remove oxygen from a closed container,
usually for anaerobiosis Haemagglutination plate for viral culture
detection Hungate Anaerobic tubes for culturing of anaerobic
microbes Incubator used for bacterial or fungal cultures
Inoculation loop: used to inoculate test samples into culture
media for bacterial or fungal cultures, antibiograms, etc. Sterilized
by passing through a blue flame. Laminar flow cabinet used to
work aseptic Latex agglutination tiles for serological analysis
Lovibond comparator a type of a colorimeter McCartney's bottle
or Flat medical bottle for simultaneous solid and liquid cultures.
McIntosh and Filde's anaerobic jar production of anaerobic
conditions for organisms that die in the presence of even little
oxygen (anaerobiosis), e.g. tetanus bacteria Microtitre plates for
ELISA
Nichrome wire loop
used to inoculate test samples into culture media for bacterial or
fungal cultures, antibiograms, etc.; sterilized by flaming to red hot
before use Petri dish/agar plate to act as a supporting container to
hold the culture medium in
Platinum wire loop
used to inoculate test samples into culture media for bacterial or
fungal cultures, antibiograms, etc.; sterilized by flaming to red hot
before use
Pre-sterilized disposable container
Pre-sterilized disposable swabs / NIH swab / postnasal swap
Roux culture bottle
Bottle designed to use laying flat, useful for growing mass
cultures and single or monolayer cultures.
Sterile loops used to inoculate test samples into culture media for
bacterial or fungal cultures, antibiograms, etc.; not heated before
use—these are disposable pre-sterilized
Thermal cycler
used to amplify segments of DNA via the polymerase chain
reaction (PCR) process.
Tissue culture bottles
to grow or keep alive cells or tissue from a living organism, e.g.
stem cells Tuberculin syringe as a normal syringe or to perform
Mantoux test
Universal container
a cylindrical small glass bottle with a screw cap used as a culture
medium holder
Vaccine bath
used to heat vaccine containing medium gently to around 45-55
degrees Celsius during vaccine production
Microscope
to observe microscopic specimens that cannot be seen by the
naked eye.
Vacuum pump
to draw out the air from any closed chamber before pumping back
CO2, O2 or N2, usually for anaerobiosis VDRL rotator for VDRL
test Specimen Dish used to hold specimen or samples.
QUESTION NO. 5
DRY AND MOIST HEAT STERILIZATION

There are different processes available through which you can

sterilize different materials. To be able to choose the right
sterilization process, one must know what method suits the
material in question more. You can’t sterilize materials that are
heat sensitive with autoclaves. Similarly, you can’t use filter
sterilization with chemicals having a greater microbe size when
compared with the membrane pores of the filter.

So, to use the dry and the moist heat sterilization in an effective
manner, you must understand the difference between both of
them.

Moist Heat Sterilization

Water at high pressure level is used in moist heat sterilization.

Autoclave is the instrument in which this process is carried out.
The temperature of the steam in this method is lower when
compared with dry heat sterilization, but the high pressure helps
with effective sterilization to take place.

The structural proteins and the organism’s enzymes are

destroyed through moist heat. This results in the death of the
organisms. Moist heat method is used for heat sensitive materials
and materials through which steam is permeable. Culture media
is also sterilized through moist heat sterilization.

Through moist heat sterilization, the most resistant of the spores

require a temperature of 121°C for around half an hour. It is a
more effective method when compared with dry heat sterilization.
This can be supported by the fact that through moist heat,
sterilization can be achieved at lower temperatures in a shorter
duration.

These were the main difference between the dry and the moist
heat sterilization methods. If you want to sterilize materials that
are more heat sensitive when compared with both these methods,
you should go for filter sterilization or chemical decontamination
methods.

Dry Heat Sterilization

In dry heat sterilization, dry heat is used for sterilizing different

materials. Heated air or fire is used in this process. As compared
to the moist heat sterilization, the temperature in this method is
higher. The temperature is usually higher than 356° F or 180 °C.

Dry heat helps kill the organisms using the destructive oxidation
method. This helps destroy large contaminating bio-molecules
such as proteins. The essential cell constituents are destroyed
and the organism dies. The temperature is maintained for almost
an hour to kill the most difficult of the resistant spores.

Things such as glassware, metal instruments, paper wrapped

things and syringes are effectively sterilized through dry heat. The
materials used in these things are heat resistant or it can be said
that they are heat stable. Powders impermeable to moisture and
anhydrous oils and fats can also be sterilized using dry heat
sterilization.