ldp2014

Bio Sci 106N (LEC):

Microbiology
and
Parasitology
(MIDTERM NOTES)
_______________________
Lennon D. Ponta-oy,
RMT
COVERAGE:
Unit 4. Microbial Genetics
Unit 5. Control of Microbial Growth
Unit 6. Drugs, Microbes, Host the
Elements of Chemotherapy
Unit 7. Infection, Infectious
Diseases and Epidemiology
Unit 8. Principles of Immunology

UNIT 4. MICROBIAL GENETICS

GENETICS

Study of the inheritance of biological

characteristics by living things (heredity)
Examines:
o The transmission of biological properties
(traits) from parent to offspring
o The expression and variation of those
traits
o The structure and function of the genetic
material
o How this material changes or evolves
Several levels:
o Organismal genetics observes the
transmission and expression of genetic
factors in the whole organism or cell
o Chromosomal genetics examines the
characteristics and actions of
chromosomes
o Molecular genetics deals with the
biochemistry of gene function

LEVELS OF STRUCTURE & FUNCTION OF

THE GENOME

GENOME sum total of genetic material carried within

a cell
Includes the chromosomes and the plasmids
CHROMOSOME discrete cellular structure composed
of a neatly packaged DNA molecule;
contains the gene
GENE

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in classic genetics, it refers to the fundamental

unit of heredity responsible for a given trait in an
organism
in the molecular and biochemical sense, it is a
portion of the chromosome that provides
information for a given cell function
specific segment of DNA (or RNA in some viruses)
that contains the necessary codes for functional
products
directs all functions of the cell, providing it with its
own particular traits and individuality
falls into three categories:
Structural Genes code for proteins
Genes that code for the RNA
Regulatory Genes control gene expression

In prokaryotes, the circular chromosome is

packaged by the action of topoisomerase
(specifically DNA gyrase).
o Bacteria package DNA in bacterial
chromosomes (single circular DNA) and
plasmids.
In eukaryotes, more complex:
o Formation of nucleosomes (linear DNA +
histone)
o Folding of nucleosomes in a spiral
formation
o Supercoiling of the spiral to form an even
greater spiral
Eukaryotes package DNA in chromatin and
chromosomes.

GENOTYPE Also the genome

Complete collection of genes
Genetic makeup, the information that
codes for all the particular characteristics
of an organism
PHENOTYPE all the physical traits or characteristics
of an organism
PHENOTYPE is the manifestation of GENOTYPE.
CONSTITUTIVE GENES genes that are expressed at all
times
INDUCIBLE GENES expressed only when needed

PACKAGING OF DNA

Packing the mass of DNA into the cell involves

compacting the DNA molecule by means of
supercoils or superhelices.

STRUCTURE & COMPOSITION OF THE

GENETIC MATERIAL

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NUCLEOTIDES building blocks of nucleic acids:
DNA (deoxyribonucleic acid) and RNA
(ribonucleic acid)
Consist of three subunits:
o Nitrogenous Bases
Purines have two rings
Adenine (A)
Guanine (G)
Pyrimidines have single ring
Thymine (T) (in DNA)
Cytosine (C)
Uracil (U) (in RNA)
o Pentose (5-Carbon sugar)
Ribose (in RNA)
Deoxyribose (in DNA)
o Phosphate group
3 Parts to Every
Nucleotide
1) Nitrogen Base

2) Pentose
3) Phosphate
Group

Four DNA
Nucleotides
Adenine (A)
Guanine (G)
Cytosine (C)
Thymine (T)
Deoxyribose
Phosphate
Group

Four RNA
Nucleotides
Adenine (A)
Guanine (G)
Cytosine (C)
Uracil (U)
Ribose
Phosphate
Group

DNA STRUCTURE
Double-stranded helix
Sugar-phosphate backbone: each deoxyribose
sugar bonds covalently in a repeating pattern with
two phosphates
Complementary base pairs (in DNA: A and T, G and
C) are held together by hydrogen bonds.
Chargaffs Rule
The number of purines is equal to the
number of pyrimidines (A=T and G=C, thus
A+G=C+T)
Antiparallel arrangement: one side of the helix
runs in the opposite direction of the other (one
helix runs from 5 to 3, the other helix runs from
3 to 5)

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NOTE:
Adenine (A) forms two H-bonds with thymine
(T).
Cytosine (C) forms three H-bonds with guanine
(G).
The bases are attracted to each other in this
pattern because each has a complementary
three-dimensional shape that fits together with
its pair.
Johann Friedrich Miester discovered DNA
(nuclein)
James Watson and Francis Crick discovered the
double-helix
structure of DNA
Oswald Avery, Colin MacLeod and Maclyn
McCarty
purified DNA and demonstrated that it
was indeed the blueprint for life.
Erwin Chargaff Chargaffs Rule

THE SIGNIFICANCE OF THE DNA

STRUCTURE

The arrangement of the N-bases has two essential

effects:
1. Maintenance of the genetic message during
reproduction
The constancy of base-pairing guarantees
that the correct order of the DNA bases will
be retained during cell division.
When the two strands are separated, each
one provides a template (pattern or model)
for the replication (exact copying) of a new
molecule.

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Because the sequence of one strand

provides the correct pattern for its
complementary strand, the codes can be
duplicated with accuracy.
2. Providing variety
The order of bases along the length of the
DNA strand provides information needed to
produce RNA and protein molecules, which
in turn are responsible for the phenotype of
the cell.
RNA STRUCTURE
It is a single-stranded molecule that can assume
secondary and tertiary levels of complexity due
to bonds within the molecule, leading to
specialized forms of RNA (mRNA, tRNA, and
rRNA).
RNA contains uracil, instead of thymine, as the
complementary base-pairing mate for adenine.
This does not change the inherent DNA code in
any way because the uracil still follows the
pairing rules.
Although RNA, like DNA, is structured with a
backbone of alternating sugar and phosphate
molecules, the sugar in RNA is ribose rather
than deoxyribose.

DNA REPLICATION

One parental double-stranded DNA molecule

is converted to two identical daughter
molecules
Requires unwinding the double helix and
exposing the nucleotides to serve as templates
for synthesis of 2 identical molecules by DNA
polymerase.

Is semiconservative (each new double-stranded

DNA molecule contains one original or
conserved strand and one new strand) and
proceeds in the 5 to 3 direction of the newly
synthesized DNA .
Replication fork point in the DNA molecule at w/c
replication occurs
Origin of Replication site that serves as the site
where replication will be initiated
Some Enzymes Involved in DNA Replication and their
Functions
Enzyme
Function
Helicase
Unzipping the DNA helix
Primase
Synthesizing an RNA
primer
DNA Polymerase III
Adding bases to the new
DNA chain; proofreading
the chain for mistakes
DNA Polymerase I
Removing RNA primers,
replacing gaps between
Okazaki fragments with
correct nucleotides,
repairing mismatched
bases
Ligase
Final binding of nicks in
DNA during synthesis and
repair
Gyrase
Supercoiling

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DNA replication by some bacteria, such as E. coli,

goes bidirectionally around the chromosome.

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THE CENTRAL DOGMA

Proposed by Francis Crick in 1957 to explain the

flow of genetic information within the cell
Also known as the one gene-one protein
hypothesis
States that:
o The genetic information contained in one
gene of a DNA molecule is used to make
one molecule of mRNA by a process
known as transcription.
o The genetic information in that mRNA
molecule is then used to make one
protein by a process known as
transcription.

make them. This blueprint exists in the

order of triplets along the DNA strands.
The order of triplets directs a proteins
primary structurethe order and type of
amino acids in the chainwhich
determines its characteristic shape and
function
Proteins contribute significantly to the
phenotype by functioning as enzymes
and structural molecules.

THE GENE-PROTEIN CONNECTION

The language of DNA exists in the order of

groups of three consecutive bases, or triplets,
on one DNA strand.
Each triplet represents a code for a particular
amino acid.
When the triplet code is transcribed and
translated, it dictates the type and order of
amino acids in a polypeptide (protein) chain.
Key points that connect DNA and protein
function:
o DNA is a blueprint that indicates which
kinds of proteins to make and how to

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THE GENETIC CODE

Set of rules specifying the relationship between

the sequence of bases in a DNA or mRNA
molecule and the order of amino acids in the
polypeptide chain encoded by that DNA or
mRNA.
The genetic code is a triplet code that is, a
code in which three base pairs in doublestranded DNA are required to specify each
amino acid in a polypeptide.
The genetic code is a degenerate code that is,
a given AA can be specified by more than one
nucleotide triplet.
The genetic code is nonoverlapping that is ,
each nucleotide is part of one, and only one
triplet.
Nucleotide triplets in mRNA, called CODONS, are
the actual coding units read by the translational
machinery during protein synthesis.
The genetic code is unambiguous that is,
every codon has one and only one meaning.
Start Codon (AUG) initiates the process of
protein synthesis
Stop Codon (UAA, UAG, and UGA)
instructs the cell to terminate
synthesis of the polypeptide chain

Major Types of RNA Involved

RNA type
Contains codes
for:
Messenger
Sequence of
(mRNA)
AA in proteins

in Protein Synthesis
Function
Translated
Carries the
DNA master
code to the
ribosomes

Yes

Transfer
(tRNA)

A cloverleaf
tRNA to carry
AA

Ribosomal
(rRNA)

Several large
structural RNA
molecules

Primer

An RNA that
can begin DNA
replication

Brings AA to
ribosomes
during
translation
Forms the
major parts of
the ribosomes
and involved
in protein
synthesis
Primes DNA

No

No

No

TRANSCRIPTION

It is the synthesis of an RNA molecule whose

base sequence is complementary to the base
sequence of a template DNA strand.
During transcription, an RNA molecule is
synthesized using the codes on DNA as a guide
or template.
Enzyme involved: RNA Polymerase
It proceeds in three stages:
o Initiation requires the RNA polymerase
to recognize a region on a gene called the
promoter region
o Elongation
o Termination
*promoter site determines where RNA synthesis
starts and which DNA strand is to
serve as the template strand
*Sigma factor guides the RNA polymerase to
the correct position on the
promoter
*template strand one strand of DNA that is
transcribed

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*nontemplate strand sometimes called the
sense, or coding strand because its
sequence is the same order as

mRNA (although it will have

thymine and uracil); not
transcribed

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Transcription
Sequence of Bases in the
DNA Template
A
T
G
C
C
G
A
A
T

Sequence of Bases in the

mRNA Template
U
A
C
G
G
C
U
U
A

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TRANSLATION (Protein Synthesis)

It refers to the synthesis of polypeptide chains

on ribosomes using a process that employs
mRNA to determine the AA sequence.
Decoding the language of nucleic acids and
converting that information into the language
of proteins
Language of mRNA is in the form of codons,
groups of three nucleotides, such as AUG, GGC,
or AAA.
o The sequence of codons on an mRNA
molecule determines the sequence of
amino acids that will be in the protein
being synthesized.
o Each codon codes for a particular amino
acid. This is the genetic code.
o Of the 64 codons, 61 are sense codons,
and 3 are nonsense codons.
o Sense codons code for AA
o Nonsense (stop) codons do not code
for AA; signal the end of the protein
molecules synthesis (UAA, UAG, UGA)
o Start codon initiates protein synthesis;
AUG codes for formylmethionine in
bacteria (in eukaryotes, methionine)
The codons of an mRNA are read sequentially;
and, in response to each codon, the appropriate
amino acid is assembled into a growing chain.
The three-base sequence of the codon
determines which tRNA brings its specific AA to
the ribosome, because the tRNA molecule
contains an anticodon (a 3-base sequence
complementary to the code of the mRNA).
Key Components:

rRNA component of the ribosome helps

position the mRNA and catalyzes peptide
bond formation.
o tRNA molecules align AA in the correct
order along the mRNA template
o aminoacyl-tRNA synthetases attach AA
to their appropriate tRNA molecules
o mRNA molecules encode the AA
sequence information for the
polypeptides being synthesized
o protein factors- facilitate several steps in
the translation process
Mechanism:
o Translation involves initiation, elongation
and termination stages.
o During the initiation stage, initiation
factors trigger the assembly of mRNA,
ribosomal subunits and initiator
aminoacyl tRNA into an initiation
complex.
o Chain elongation involves sequential
cycles of aminoacyl tRNA binding, peptide
bond formation, and translocation
(enzyme-directed shifting of the ribosome
to the next position on the mRNA strand,
which causes the blank tRNA to be
discharged from the ribosome at the E
site). The net result is that aminoacyl
tRNAs add their AA to the growing
polypeptide chain in an order specified by
the codon sequence in mRNA.
o Chain termination occurs when a stop
codon in mRNA is recognized by release
factors, which cause the mRNA and newly
o

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formed polypeptide to be released from
the ribosome.
Example:
DNA
template
a.

mRNA
(codon)

tRNA
(anticodon)

Amino Acid

G
G
C

C
C
G

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G
G
C

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Proline

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contain genes coding for anabolic
enzymes

MUTATION

GENETIC REGULATION OF
PROTEIN
SYNTHESIS AND METABOLISM
Control mechanism ensures that genes are
active only when their products are required.
Major form of gene regulation in prokaryotes is
through systems called operons
Operon section of DNA that contains one or
more structural genes along with a
corresponding operator gene that controls
transcription
o Inducible operons - the operon is turned
on (induced) by the substrate of the
enzyme for which the structural genes
code; e.g., catabolic operons, lactose (lac)
operon in bacteria
o Repressible operons -several genes in
series are turned off (repressed) by the
product synthesized by the enzyme; often

Changes in the base sequence of a DNA

molecule
It can involve the loss of base pairs, the addition
of base pairs, or a rearrangement in the order of
base pairs.
Wild type/strain - A microorganism
that exhibits a natural, nonmutated
characteristic
Mutant strain A microorganism that
bears a mutation
- show variance in morphology,
nutritional characteristics, genetic
control mechanisms, resistance to
chemicals, temperature preference,
and nearly any type of enzymatic
function
- very useful for tracking genetic
events, unraveling genetic
organization, and pinpointing genetic
markers

SPONTANEOUS MUTATION
random change in the DNA arising from errors in
replication that occur without a known cause
INDUCIBLE MUTATION
result from exposure to known mutagens, which
are primarily physical or chemical agents that
damage DNA and interfere with its functioning
Selected Mutagenic Agents and Their
Effects
Agent
Effect

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Chemical
Nitrous acid,
bisulfite

Remove an
amino group
from some
bases
Ethidium
Inserts
bromide
between the
paired bases
Acridine dyes
Cause
frameshifts
due to
insertion
between
base pairs
Nitrogen
Compete
base analogs
with natural
bases for
sites on
replicating
DNA
Physical (primarily types of radiation)
Ionizing
Form free
(gamma
radicals that
rays, X rays)
cause single
or double
breaks in
DNA
Ultraviolet
Causes
cross-links
between
adjacent
pyrimidines

Categories of Mutations:
Point (base) mutations

Involve addition, substitution or deletion

of bases
Missense mutation
o If the base substitution results in an
amino acid substitution in the synthesized
protein
Nonsense mutation
o changes a normal codon into a stop
codon that does not code for an amino
acid and stops the production of the
protein wherever it occurs
Silent mutation
o alters a base but does not change the
amino acid and thus has no effect
Back-mutation
o occurs when a gene that has undergone
mutation reverses (mutates back) to its
original base composition
Frameshift mutation
o occurs when one or more bases are
inserted into or deleted from a newly
synthesized DNA strand
o so named because the reading frame
(specific order of nucleotides or codons
provided by the mRNA formed during
transcription) has been changed

Repair of Mutations:
Proofreading mechanism of the DNA
Photoactivation or light repair
o Fix DNA damaged by UV radiation
o requires visible light and a light-sensitive
enzyme, DNA photolyase, which can
attach to sites of abnormal pyrimidine
o

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bonding and restore the original DNA

structure
Excision repair
o Mutations can be excised by a series of
enzymes that remove the incorrect base
and add the correct ones.
Mismatch Repair
o Targets errors made during DNA
replication, when improperly base-paired
nucleotides sometimes escape the normal
proofreading mechanisms
o Because mismatched base pairs do no
hydrogen-bond properly, their presence
can be detected and corrected

Ames Test
A mutant strain of Salmonella is used
to learn whether a particular
chemical (e.g., a food additive or a
chemical used in some type of
cosmetic product) is a mutagen
If exposure to the chemical causes a
reversal of the organisms mutation
(back mutation), then the chemical
has been shown to be mutagenic.
If the chemical is mutagenic, then it
might alse be carcinogenic (cancercausing) and should be tested using
laboratory animals or cell cultures.

BACTERIAL RECOMBINATION

An event in which one bacterium donates DNA

to another bacterium

End result of which is a new strain different from

both the donor and the original recipient strain
In general, any organism that has acquired
genes that originated in another organism is
called a recombinant
Depends largely on the bacterias extreme
versatility in acquiring and expressing the
genetic material of other bacteria and even
other organisms
Are generally beneficial: They can provide
additional genes for resistance to drugs and
metabolic poisons, new nutritional and
metabolic schemes, increased virulence, and
adaptations to changing environmental
conditions

Ways in Which Bacteria Acquire New

Genetic Information
1. Mutations involve changes in the base
sequence of genes
2. Lysogenic Conversion
Involves bacteriophages and the
acquisition of new viral genes
While the prophage is integrated into the
bacterial chromosome, the bacterial cell
can produce gene products that are
coded for by the prophage genes.
3. Transduction
Means to carry across
Involves bacteriophages and the
acquisition of new bacterial genes
Process by which a bacteriophage serves
as the carrier of DNA from a donor cell to
a recipient cell

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Although it occurs naturally in a broad

spectrum of bacteria, the participating
bacteria in a single transduction event
must be the same species because of the
specificity of viruses for host cells
Only small segments of DNA are
transferred from cell to cell

4. Transformation

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Involves the uptake of naked DNA

This nonspecific acceptance by a

bacterial cell of small fragments of
soluble DNA from the surrounding
environment.
Cells that are capable of accepting
genetic material through this means
are termed competent.

5. Conjugation
Involves the transfer of genetic
information from one cell to another
through a hollow sex pilus
Discovered by Joshua Lederberg and
Edward Tatum in 1946 while
experimenting with E. coli.

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Involves a specialized type of pilus

called the sex pilus (F pilus or
conjugation bridge)
A bacterial cell (donor cell or F+ cell)
possessing a sex pilus attached by
means of the sex pilus to another
bacterial cell (recipient or F- cell).
Some genetic material (plasmid) is
then transferred through the hollow
sex pilus from the donor cell to the
recipient cell.
Has great biomedical importance:
o Special resistance (R)
plasmids, or factors, that bear
genes for resisting antibiotics
and other drugs are commonly
shared among bacteria through
conjugation. Transfer of R
factors can confer multiple
resistance to antibiotics such as
tetracycline, chloramphenicol,
sulfonamides, and penicillin.
o Other types of R factors carry
genetic codes for resistance to
heavy metals (nickel and
mercury) or for synthesizing
virulence factors (toxins,
enzymes, and adhesion
molecules) that increase the
pathogenicity of the bacterial
strain.

GENETIC ENGINEERING

Manipulation of an organisms genome and is

often used in conjunction with biotechnology,
the use of an organisms biochemical and
metabolic pathways for industrial production of
proteins
Plasmids are frequently used as vectors or
vehicles for inserting genes into cells
Bacteria, yeasts, human leukocytes,
macrophages, and fibroblasts have been used

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as genetically engineered manufacturing

plants for proteins such as human growth
hormone (somatotropin), somatostatin,
plasminogen-activating factor, insulin and
interferon.
Applications:
o Therapeutic applications
Synthetic genes linked to the galactosidase gene (lacZ) in a
plasmid vector were inserted into
E. coli, allowing E. coli to produce
and secrete the two polypeptides
used to make human insulin
Cells and viruses can be modified
to produce a pathogens surface
protein, which can be used as a
vaccine.
DNA vaccines consist of
recombinant DNA cloned in
bacteria.
Gene therapy can be used to cure
genetic diseases by replacing the
defective or missing gene.
o Genome Projects
Nucleotide sequences of the
genomes over 1000 organisms,
including humans, have been
completed.
This leads to determining the
proteins produced in a cell.
o Scientific Applications
DNA can be used to increase
understanding of DNA, for genetic
fingerprinting, and for gene
therapy

DNA sequencing machines are

used to determine the nucleotide
base sequence of restriction
fragments in shotgun sequencing.
Bioinformatics is the use of
computer applications to study
genetic data; proteomics is the
study of a cells proteins
DNA probes can be used to quickly
identify a pathogen in body tissue
or food
Forensic microbiologists use DNA
fingerprinting to identify the source
of bacterial or viral pathogens
Bacteria may be used to make
nano-sized materials for
nanotechnology machines.
Agricultural Applications
Cells from plants with desirable
characteristics can be cloned to
produce many identical cells. These
cells can then be used to produce
whole plants from which seeds can
be harvested.
Incorporating nitrogen-fixing
capabilities into additional soil
microorganisms
Make plants that are resistant to
insects, as well as to bacterial and
fungal diseases.
Increase the size and nutritional
value of foods.

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UNIT 5. CONTROL OF MICROBIAL

GROWTH

Definition of Terms:
Sterile
Free of life of every kind
Sterilization
Complete destruction or removal of all forms
of microbial life including endospore.
Usually done by steam under pressure,
sterilizing gas, or ethylene oxide.
Disinfection
Destruction of vegetative pathogens on
inanimate objects
Partial destruction through physical and
chemical methods
Antisepsis
Destruction of vegetative pathogens on living
tissue
Treatment is almost always by chemical
antimicrobials

Sanitization
Treatment intended to lower microbial counts
on eating and drinking utensils to safe public
health levels.
May be done with high temperature washing
or by dipping into a chemical disinfectant.
Degermining
Removal of microbes from a limited area,
such as the skin around an injection site.
Mostly mechanical removal by an alcoholsoaked swab
Microbicidal
Property of destroying microbes
Bactericidal, fungicidal, etc.
Microbiostatic
Property of inhibiting microbial growth and
multiplication
Bacteriostatic, fungistatic, etc.
Disinfectant
Chemical agents applied on inanimate
surface, too toxic to be applied on living
tissues
Antiseptic
Chemical agents which could have a
cidal or static effect, applied topically on
living tissues.
Septic
Condition characterized by presence of
pathogens (particularly on living tissues)
Aseptic
Condition characterized by the absence of
pathogens
Thermal death time

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Minimum time required to kill suspension
of microbes in a given temperature in a
specified environment
Thermal death point
Maximum temperature in a given time to
destroy all microbes present

Several Factors that Influence the

Rate at which Antimicrobial Agents
Work:
Exposure time of the agent
Numbers of microbes present
Relative resistance of microbes (for example,
endospores versus vegetative forms)
Activity of the agent (microbicidal versus
microbiostatic)

2 General Methods of Microbial

Growth
A. Physical methods - control is achieved by
modifying environmental condtions
a. Scrubbing with soap and water
Washing based on standard operating
procedure before and after an activity
b. Filtration
Used to remove microbes for liquids
or gases, with use of filters 0.2 to
0.45m pore.
Filters most bacteria, while viruses
and small bacteria pass through
them.
For antibiotic solutions, CHO
solutions, vaccines, culture media
which cannot be heated

Liquid: pulling solution through

cellulose acetate or cellulose nitrate
medium with vacuum
Air: HEPA filters (removes organisms
> 0.3m)
c. Sedimentation
Allowing solid or solutes or particulate
matter to settle at bottom of liquid
d. High temperature
i. Pasteurization
Use brief exposures to
moderately high temp to reduce
& eliminate pathogens, w/o
eliminating viable beneficial
microbes and w/o altering
chemical nature of food.
Kills non-sporeforming
organisms in heat sensitive
materials (milk products, wine,
etc.)
Batch/LTH 63C for 30 mins
Flash/HTST 72C for 15 secs
ii. Moist Heat
Denatures bacterial proteins;
water hastens breaking of Hbonds which hold proteins in
their 3 dimensional
configurations; more
penetrating
Boiling
100C for 15 mins or 30
mins
Kills vegetative cells but
not spores

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For sterilization of
apparatus that are heat
resistant
Suitable in situations from
which endospores and
hepatitis virus are known
to be absent
Autoclaving
Steam under pressure
Most effective method
121C at 15lbs psi for
15mins
Destroys spores
For sterilization of culture
media and surgical
instruments
Biologic indicator: Bacillus
stearothermophilus
Tyndallization
Also
Fractional/Discontinuous/
Intermittent Sterilization
For heat labile, sporecontaining material
100C for 30 mins for 3
consecutive days; 60C for
1hr for 5-6 days
Inspissation
75C - 80C for 2 hrs for 3
consecutive days
Principle: Thickening
through evaporation
For high protein content
media

e. Dry Heat
Denatures proteins; kills organisms
by oxidation; requires higher
temperature and long exposure to
ensure complete sterilization
Passing through a flame
For loops needles, mouth
of tubes as well as plates
Hot Oven
For sterilization of
glasswares
160C - 170C for 1.5 to 2
hrs
Incineration
For infectious wastes;
burning wastes into ashes
870C - 980C (800C to
6500C)
Banned in the Philippines
Cremation
Burning dead bodies (with
communicable disease) to
ashes
f. Low Temperature
Limits rate of microbial
reproduction
Microbiostatic
Commonly used to preserve food,
media, and cultures
5C for refrigeration temperature;
0C or subzero; freeze drying
through sublimation
g. Radiation
Ionizing Radiation

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ldp2014
Short wavelength, high
energy gamma rays and
x-rays
For plastic syringes,
catheter or gloves
Has deep penetrating
power and works by
causing breaks in the DNA
of target organisms
Non-ionzing Radiation
Long wavelength low E UV
light
Very little penetrating
power
Works by creating dimers
between adjacent
pyrimidines, which
interferes with replication
B. Chemical Methods
a. Halogens
Chemicals based on elements from
group VII of the periodic table
i. Chlorine
Kills microbe by disrupting the
plasma membrane
Chlorine gas, hypochlorites,
chloramines all work by
disrupting disulfide bonds
Na hypochlorite or bleach
(1:10)
ii. Iodine
Reactive by precipitating
proteins and oxidizes essential
enzymes
I2 + detergent = iodophor

I2 in alcohol = I2 tincture
iii. Fluorides
In toothpaste, H2O supply
b. Phenolic Compounds
Act by disrupting lipid containing
membranes, resulting in leakage of
cellular contents
Carbolic acid, Lysol, cresol-opolyphenol
Standard disinfectant in the lab
c. Detergents
i. Anionic Detergents (Soaps)
Removing grease and soil that
contains microbes
ii. Quaternary Ammonium Compounds
(QUATS)
Cationic detergents
Act as surfactants that alter the
membrane permeability of
some bacteria and fungi
Benzalkonium chloride
(Zephiran)
For digestion and
decontamination of sputum
Inactivated by ORGANIC
SUBSTS
Disadvantage: Nonsporicidal;
Nontuberculoidal
d. Alcohols
Most effective and most used
Act as surfactants, dissolving
membrane lipids and
coagulating proteins of

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ldp2014
vegetative bacterial cells and
fungi
Non-sporicidal
Used on skin (as antiseptic) and
on thermometers and injection
vial rubber septum (as
disinfectant)
Evaporates easily
i. Ethanol
70% is more effective than
95% EtOH
70% EtOH kills nearly 90% of
cutaneous microbiota w/in 2
mins
ii. Isopropanol
Highest bactericidal activity at
70-80%
Less activity against
endospores, fungi and viruses
e. Aldehydes
Denature proteins and DNA by
alkylation
Used for disinfecting surgical
instruments
Formaldehyde and
Glutaraldehyde (pH 7.5 kills
Staphylococci in 5 mins, tubercle
bacilli in 10 mins and endospore
for 12 mins
f. Acids destroy or inhibit microbial cells
i. Organic Acids
widely used in food preservation
because they prevent spore
germination and bacterial and
fungal growth and because they

are generally regarded as safe to

eat
Acetic acid, Lactic acid, Benzoic
acid, Sorbic acid
Vinegar (household disinfectant)
g. Alkalis inactivates proteins
i. Ammonium hydroxide
`reliably destroys prions
h. Dyes
antimicrobial effects are apparently
due to the way they insert into
nucleic acids and cause mutations
interfere with cell wall synthesis
i. Hydrogen peroxide
produces highly reactive hydroxyl
free radicals that damage protein
and DNA while also decomposing
to O2 gas, which is toxic to
anaerobes
3 to 6% solutions antiseptic
6 to 25% solutions for sterilization
mixtures, disinfects implants,
prostheses and contact lenses
Strong solutions are sporicidal
j. Ethylene oxide
Colorless gas, soluble in water and
organic solvents used for heat
sensitive items
Used for sterilizing aircrafts for
space explorations
Can be used for sterilizing
disposable utensils, instruments
and prostheses
k. Ozone

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ldp2014
Strong oxidizing agent
Oxidizes cellular biochemical
Disinfect drinking water
l. Heavy Metals
Acts with sulfhydryl groups of
proteins, thus inactivates them
Inactivated by organic materials
(e.g., blood)
Mercury-containing compound is no
longer recommended toxic to the
environment
Silver Nitrate eyedrop used to
prevent N. gonorrhoeae infection in
newborns
m. Chlorhexidine
surfactant and protein denaturant
with broad microbicidal properties,
although it is not sporicidal
Solutions of chlorhexidine are used
as skin degerming agents for
preoperative scrubs, skin cleaning,
and burns
n. Antibiotics
Antimicrobial substances produced
by microbes, used for treating
humans and animals
Modes of action: inhibits CW
synthesis, inhibits CM function,
inhibits protein synthesis, inhibits
nucleic acid synthesis

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