Microbial World and You

Chapter 1
What is Microbiology?
Micro - too small to be seen with the naked
eye
Bio - life
ology - study of
Organisms included in the study
of Microbiology
1. Bacteria Bacteriology
2. Protozoans Protozoology
3. Algae Phycology
4. Parasites Parasitology
5. Yeasts and Molds
• Fungi Mycology
6. Viruses Virology

Microorganisms - Microbes - Germs

5 Kingdoms of Living Organisms
1. Animalia
2. Plantae
3. Fungi
4. Protista
5. Monera - Bacteria and Cyanobacteria

Eukaryotic vs. Prokaryotic

5 Characteristics of Life
1. Cells
2. Maintain structure by taking up
chemicals and energy from the environment
3. Respond to stimuli in the external
environment
4. Reproduce and pass on their organization
to their offspring
5. Evolve and adapt to the environment
Taxonomic Classification
Man
Kingdom Animalia
Phylum Chordata
Class Mammalia
Order Primate
Family Hominidae
Genus Homo
species sapien
Taxonomic Classification
Cat
Kingdom Animalia
Phylum Chordate
Class Mammalia
Order Carnivora
Family Felidae
Genus Felis
species domestica
Binomial System of Taxonomic
Classification
Use only the Genus and species
Homo sapien
Felis domestica
Escherichia coli
Genus and species are either underlined or
italicized
Genus is always capitilized
species is never capitilized
Classification System
3 Domains 1978 Carl Woese
• 1. Bacteria
• Unicellular prokaryotes with cell wall containing
peptidoglycan
• 2. Archaea
• Unicellular prokaryotes with no peptodoglycan in
cell wall
• 3. Eukarya
• Protista
• Fungi
• Plantae
• Animalia
Bacteria - what comes to mind?
Diseases
Infections
Epidemics
Food Spoilage
Only 1% of all known bacteria cause human
diseases
About 4% of all known bacteria cause plant
diseases
95% of known bacteria are non-pathogens
Microbes Benefit Humans
1.Bacteria are primary decomposers -
recycle nutrients back into the environment
(sewage treatment plants)
2. Microbes produce various food products
• cheese, pickles, sauerkraut, green olives
• yogurt, soy sauce, vinegar, bread
• Beer, Wine, Alcohol
3. Microbes are used to produce Antibiotics

Penicillin

Mold
• Penicillium notatum

1928 Alexander Fleming

4. Bacteria synthesize chemicals that our
body needs, but cannot synthesize
Example: E. coli
• B vitamins - for metabolism
• Vitamin K - blood clotting

Escherichia coli
• Dr. Escherich
• Colon (intestine)
5. Biochemistry and Metabolism
Very simple structure
rapid rate of reproduction
provides “instant” data
6. Microbial Antagonism
Our normal microbial flora prevents
potential pathogens from gaining access to
our body
7. Insect Pest Control
Using bacteria to control the growth of
insects

Bacillus thuringiensis
• caterpillars
• bollworms
• corn borers
8. Bioremediation
Using microbes to clean up pollutants and
toxic wastes
Exxon Valdez - 1989

2 Genera
• Pseudomonas sp.
• Bacillus sp.
9. Recombinant DNA Technology
Gene Therapy
Genetic Engineering

Bacteria can be manipulated to produce

enzymes and proteins they normally would
not produce
• Insulin
• Human Growth Hormone
• Interferon
10. Microbes form the basis of
the food chain
Marine and fresh water microorganisms
Microbes do benefit us, but they
are also capable of causing many
diseases
Pneumonia Whooping Cough
Botulism Typhoid Fever Measles
Cholera Scarlet Fever Mumps
Syphilis Gonorrhea Herpes 1
Chlamydia Tuberculosis Herpes 2
Meningitis Tetanus RMSV
Strep Throat Lyme Disease AIDS
History of the Study of
Microorganisms
1665 Robert Hooke
• “little boxes” - “cells”
• Cell Theory - all living things are made up of
cells
Anton van Leeuwenhoek 1674
- 1st person to actually see living microorganisms

“wee animalcules”
Spontaneous Generation
Theory that life just “spontaneously”
developed from non-living matter

Example:
• toads, snakes and mice - moist soil
• flies and maggots - manure and decaying flesh
Experiments to disprove
Spontaneous Generation
Francesco Redi 1668

Rudolph Virchow 1858

• Theory of Biogenesis
• Cells can only arise from preexisting cells

Louis Pasteur 1861

Pasteur designed special “swan-necked flasks”
with a boiled meat infusion

Shape of flask allowed air in (vital force) but trapped

dust particles which may contain microbes
Germ Theory of Disease
Hard for people to believe that diseases
were caused by tiny invisible “wee
animalcules”
Diseases, they thought, were caused by:
• demons
• witchcraft
• bad luck
• the wrath of God
• curses
• evil spirits
Robert Koch - 1st to prove that
bacteria actually caused diseases
1876
Microbial Etiology of Infectious Disease
• etiology - the cause of a disease
Established “scientific rules” to show a
cause and effect relationship between a
microbe and a disease
• Koch’s Postulates
Koch’s Postulates
1. The same organisms must be found in all
cases of a given disease.
2. The organism must be isolated and grown
in pure culture.
3. The isolated organism must reproduce
the same disease when inoculated into a
healthy susceptible animal.
4. The original organism must again be
isolated from the experimentally infected
animal.
Exceptions to Koch’s Postulates
1. Some organisms have never been grown in
pure culture on artificial media

Treponema pallidum - Syphilis

 Exceptions to Koch’s Postulates

Mycobacterium leprae
Leprosy

Never been grown in pure culture on artificial media

Abdominal cavity of the Seven Banded Armadillo

Exceptions to Koch’s Postulates
In exclusively human diseases, it is not
morally acceptable to inoculate a deadly
pathogen into a “human guinea pig”

HIV
Koch established the Microbial
Etiology of 3 important diseases
of his day
1. Cholera (fecal-oral disease)
• Vibrio cholerae
2. Tuberculosis (pulmonary infection)
• Mycobacterium tuberculosis
3. Anthrax (sheep and cattle)
• Bacillus anthracis
Anthrax
Bacillus anthracis
• Gram (+), non-motile, aerobic, spore forming rod
• Streptobacilli with central spores
• Livestock
• Sheep, cattle, goats
• Humans
• Handle hides, wool, goat hair, handicrafts from the Middle
East made from animal products
 3 Forms of Human Anthrax

1. Cutaneous Anthrax
• Enters thru cut or
abrasion
• Results in painless
ulcer (1-3 cm) with
black (necrotic) center
• About 20% mortality
rate in untreated cases
2. Gastrointestinal Anthrax
• Contaminated meat
• Abdominal pain, fever,
vomiting blood, severe
diarrhea
• 25% to 60% mortality rate
3. Inhalation Anthrax
• Initial symptoms
resemble common cold
• Progress to severe
breathing problems and
shock
• Usually results in death
1-2 days after onset of
acute symptoms
• Mortality rate 99% in
untreated cases
• Treatment usually not
effective after
symptoms are present
Anthrax as a Biological Weapon
Deadly if not treated early
Spores can be produced in large quantities using basic
knowledge of biology
Spores may remain viable for years (60 at least)
Spores can be spread
• Missiles, rockets, bombs, mail, crop dusters ?
No cloud or color
No smell
No taste
Antibiotics – only effective if administered early (within
24 –48 hours)
Golden Age of Microbiology 1857 - 1914

Hooke-First observation of cells

Van Leeuwenhoek-First observation of live
microorganisms
Linaeus - Nomenclature for organisms
Pasteur
• Pasteurization
• Fermentation
• Disproved spontaneous generation
Joseph Lister
Phenol to treat surgical wounds – 1st
attempt to control infections caused
by microorganisms (aseptic surgery)
Robert Koch (Germ theory of
disease)
Koch’s Postulates
Edward Jenner
vaccination
Paul Erlich
1st synthetic drug used to treat
infections
Salvarsan - arsenic based chemical to
treat Syphilis
“salvation” from Syphilis
Naming of Bacteria
Genus and species - Binomial System of
Taxonomic Classification

Information usually given:

• 1. Describes an organism
• 2. Identifies a habitat
• 3. Honors a scientist or researcher
Bacterial Morphology
Bacilli

Cocci

Spiral
Arrangements
Staphylo
Strepto
Diplo
Sarcinae
Tetrad

Vibrio comma shaped

bacter bacilli
bacterium bacilli
Staphylococcus aureus Escherichia coli
Staphylococcus Bacillus anthrasis
epidermidis Salmonella enteridis
Streptococcus Streptococcus
pneumoniae pyogenes
Steptococcus lactis
Vibrio cholerae
Streptococcus faecalis
Rhodospirillium Erlichia canis
rubrum Campylobacter jujuni
Bacillus subtilis Helicobacter pylori
Micrococcus luteus Enterobacter
aerogenes
Definitions:
Eubacteria: with murein or peptidoglycan
on cell wall
Archaebacteria: Without murin
Fermentation: metabolic process that
converts sugar to acids, gases or alcohol
(occurs in yeast and bacteria and in oxygen
starved muscle cells.
 Ch 3:

Observing
Microorganisms
Through a
Microscope
Q&A
Acid-fast staining of a
patient’s sputum is a
rapid, reliable, and
inexpensive method to
diagnose tuberculosis.
What color would
bacterial cells appear if
the patient has
tuberculosis?
Objectives
Review the metric units of measurement
Define total magnification and resolution
Explain how electron and light microscopy differ
Differentiate between acidic and basic dyes
Compare simple, differential, and special stains
List the steps in preparing a Gram stain. Describe the
appearance of Gram-positive and Gram-negative
cells after each step
Compare and contrast Gram stain and acid-fast stain
Explain why endospore and capsule stains are used
 Units of Measurement

Review Table 3.1

• 1 µm = ______ m = ______ mm
• 1 nm = ______ m = ______ mm
• 1000 nm = ______ µm
• 0.001 µm = ______ nm
 Sizes Among Microorganisms
• Protozoa: 100 µm
Cells Alive –
• Yeasts: 8 µm How big is a . . .?

• Bacteria: 1 - 5 µm (some much longer than

wide)
• Rickettsia: 0.4 µm = _________ nm
• Chlamydia and Mycoplasma: 0.25 µm
• Viruses: 20 – 250 nm
Principles of the Compound Light
Microscope
Magnification: Ocular and
objective lenses of
compound microscope (total
mag.?)
Resolution: Ability of lens to . . .
Maximum resolving power ___ m
Contrast: Stains change refractive
index  contrast between
bacteria and surrounding medium Fig 3.1
 Refractive Index
• Measures light-bending
ability of a medium
• Light may bend in air so
much that it misses the
small high-magnification
lens.
• Immersion oil is used to
keep light from bending.
Fig 3.3
 Microscopy: The Instruments
Darkfield Microscopy
Brightfield Microscopy
• Light objects visible
• Simplest of all the against dark background.
optical microscopy • used to enhance the
illumination. techniques contrast in unstained
• Dark objects are visible samples.
against a bright • Instrument of choice for
spirochetes
background.
Spirochetes (Treponema pallidum) viewed with darkfield microscope
Fluorescence Microscopy
• Uses UV light.
• Fluorescent substances
absorb UV light and emit
visible light.
• Cells may be stained with
fluorescent chemicals
(fluorochromes).
• Immunofluorescence

Fig 3.6; T. pallidum

Figure 3.6a

Fig 3.6b
Principle of
Immunofluorescence
Electron Microscopy: Detailed Images of
Cell Parts
Uses electrons, electromagnetic lenses, and
fluorescent screens
Electron wavelength ~ 100,000 x smaller than
visible light wavelength
Specimens may be stained with heavy metal
salts
Two types of EMs:?
SEM or TEM?

Bacterial division

Leaf surface
 ?

10,000-100,000; resolution 2.5 nm.

Rod-shaped Mycobacterium avium
Preparation of Specimens for Light
Microscopy
• Staining Techniques Provide Contrast
• Smear  air-dry  heat-fix
• Basic dyes: cationic chromophore
• Acidic dyes: anionic chromophore 
negative staining (good for capsules)
• Three types of staining techniques:
Simple, differential, and special
 Simple Stains
Differential Stains
• Use a single basic
dye. React differently with
• A mordant may be different bacteria
used to hold the • Gram stain
stain or coat the
specimen to • Acid fast stain
enlarge it.
Review of different staining techniques
Important Staining Reactions in Microbiology

For Gram stain

technique compare
to Fig 3-12
Gram Stain

crystal violet

safranin
 Acid Fast Stain
• Cells that retain a basic stain in the presence of
acid-alcohol are called acid-fast.
• Non–acid-fast cells lose the primary stain when
rinsed with acid-alcohol, and are counterstained
with a different color basic stain
Special Stains See Fig 3.14

• Endospore stain: Heat is

required to drive a stain into the
endospore.

• Flagella staining: requires a

mordant to make the flagella wide
enough to see.
• Capsule stain uses basic stain
and negative stain
e.g., 1 m = 10 dm = 100 cm = 1000 mm=10-6micr=10-9nm
That is exponential notation is a power of 10, for example 6.75 x 109,
then 6.75 is called the coefficient and 9 is the power or exponent.

Total magnification we multiply objective lens magnification (power) by

the ocular lens magnification (power)
Objective lens magnification ocular lens magnification
(power) (power)
Low power 10x X 10x = 100x
High power 40x X 10x = 400x
Oil immersion 100x X 10x = 1000x

Resolution or resolving power is defined as the ability of lenses to

distinguish fine detail and structure or it refers to the ability of lenses to
distinguish two points a specified distance apart.
https://www.youtube.com/watch?v=3LIZBn7bS4s

Electron Microscopy:
The better resolution of electron microscopes is due to the shorter
wavelength of electrons about 100,000 times smaller than wavelength
of visible light.
Two types of EM:
1. Transmission electron microscope (TEM):
Produces transmission electron micrograph (resolution 10 pm and
magnify 10,000-100,000X). Uses salts of heavy metals e.g., lead,
tungsten. Used for positive staining (staining specimen) or negative
staining (staining background).
Disadvantages of TEM:
1. No three-dimensional view of examined object.
2. Specimens must be fixed. This kills the specimen and this
treatment may cause production of artifacts.

2. Scanning electron microscope (SEM):

The produced image is called scanning electron micrograph. It is useful
in study the surface structure of intact cells and viruses.
It can resolve objects as close together as 10 nm and total
magnification of 1000-10,000X.
Disadvantages of SEM
1. Compared to TEM, the SEM can maintain lower magnification
power.
2. Lower resolution powers than TEM.
Stains
Salts composed of a positive and negative ions, one of which is colored
and is known as the chromophore.
The color of basic dyes is in the positive ion
While in acidic dyes, it is in the negative ion
Bacterial cells are slightly negatively charged at pH 7
Therefore, basic dyes are used to stain the negatively charged bacterial
cell.
Basic dyes include: crystal violet, methylene blue, malachite green and
safranin.
Acidic dyes are not attracted to most bacterial cells, because the dye’s
negative ions are repelled by the negatively charged bacterial surface,
but they are useful for:
Negative staining (preparation colorless bacteria against a colored
background
Negative staining is used for observing overall cell shape, size and
capsules.
Distortion of cell size and shape are minimized, because fixing is not
necessary and the cells do not pick up the stain

Examples for acidic dyes: eosin, acid fuchsin and nigrosine.

Smear preparation link:
https://www.youtube.com/watch?v=J0LJZ2QsXPE

Differential stains
1. Gram’s stain
2. Acid Fast stain: binds to bacterial waxy material in the cell walls of
Mycobacterium e.g., M. tuberculosis and M. leprae + Nacardia
 Ch 4

Functional
Anatomy of
Prokaryotic and
Eukaryotic
Cells
 Objectives
Compare and contrast the overall cell structure of prokaryotes and
eukaryotes.
Identify the three basic shapes of bacteria.
Describe structure and function of the glycocalyx, flagella, axial filaments,
fimbriae, and pili.
Compare and contrast the cell walls of gram-positive bacteria, gram-negative
bacteria, acid-fast bacteria, and mycoplasmas.
Differentiate between protoplast, spheroplast, and L form.
Describe the structure, chemistry, and functions of the prokaryotic plasma
membrane.
Identify the functions of the nuclear area, ribosomes, and inclusions.
Describe the functions of endospores, sporulation, and endospore
germination.

What you should remember from Bio 31:

Define organelle. Describe the functions of the nucleus, endoplasmic
reticulum, ribosomes, Golgi complex, lysosomes, vacuoles, mitochondria,
chloroplasts, peroxisomes. Explain endosymbiotic theory of eukaryotic
evolution.
Comparing Prokaryotic and
Eukaryotic Cells
Common features:
➢DNA and chromosomes
➢Cell membrane
➢Cytosol and Ribosomes

Distinctive features: ?
Prokaryotes

◼ One circular chromosome,

not membrane bound
◼ No histones
◼ No organelles
◼ Peptidoglycan cell walls
◼ Binary fission
Size, Shape, and Arrangement
Average size: 0.2 -1.0 µm  2 - 8 µm
Three basic shapes
1. Bacillus, -i
2. Coccus, -i
3. Spirals (Vibrio,
Spirillum, Spirochete)

Most monomorphic, some pleomorphic

Variations in cell arrangements (esp. for
cocci) Review Figs. 4.1, 4.2, and 4.4
Spiral Bacteria

Figure 4.4
Pleomorphic
Corynebacteria

Monomorphic
E. coli
Cell Arrangement
External Structures
located outside of cell wall

◼ Glycocalyx
◼ Flagellum /-a
◼ Axial filaments
◼ Fimbria /-ae
◼ Pilus /-i
Glycocalyx
◼ Many bacteria secrete external surface layer
composed of sticky polysaccharides,
polypeptide, or both
◼ Capsule: organized and firmly attached to cell
wall
◼ Slime layer: unorganized and loosely attached
◼ Allows cells to attach
→ key to biofilms
◼ Prevents phagocytosis
→ virulence factor
◼ E.g.: B. anthracis, S. pneumoniae,
S. mutans
Flagellum – Flagella
◼ Anchored to wall and membrane
◼ Number and placement determines if atrichous,
monotrichous, lophotrichous,
amphitrichous, or peritrichous

Fig 4.7
 Flagellar Arrangement

_______

___________
Motility
◼ Due to rotation of flagella
◼ Mechanism of rotation: “Run and tumble”
◼ Move toward or away from stimuli (taxis)
◼ Chemotaxis (phototaxis and
magnetotaxis)
◼ Flagella proteins are H antigens
(e.g., E. coli O157:H7)
“Run and Tumble”

Fig 4.9
Axial Filaments Fimbriae and Pili
◼ Endoflagella ◼ Fimbriae allow
◼ In spirochetes attachment
◼ Anchored at one end ◼ Pili are used to
of a cell transfer DNA from
one cell to another
◼ Rotation causes cell
to move

Fig 4.10
 Cell Wall
◼ Rigid for shape & protection
 prevents osmotic lysis
◼ Consists of Peptidoglycan (murein) →
polymer of 2 monosaccharide subunits
 N-acetylglucosamine (NAG) and
 N-acetylmuramic acid (NAM)

◼ Linked by polypeptides (forming peptide

cross bridges) with tetrapeptide side chain
attached to NAM
◼ Fully permeable to ions, aa, and sugars
(Gram positive cell wall may regulate movement of cations)
Fig 4.13
 Gram + Gram –
Cell Wall Cell Wall
◼ Thick layer of
peptidoglycan ◼ Thin peptidoglycan
◼ Negatively charged ◼ Outer membrane
teichoic acid on ◼ Periplasmic space
surface
 Gram-Positive Cell Walls
◼ Teichoic acids
 Lipoteichoic acid links to plasma membrane
 Wall teichoic acid links to peptidoglycan

◼ May regulate movement of cations

◼ Polysaccharides provide antigenic variation

Fig.4.13b
 Gram-negative Cell Wall
Lipid A of LPS acts as endotoxin; O polysaccharides
are antigens for typing, e.g., E. coli O157:H7
Gram neg. bacteria are less sensitive to medications
because outer membrane acts as additional barrier.
LPS layer = outer layer of outer membrane

(protein rich gel-like fluid)

Fig 4.13
Gram Stain Mechanism
◼ Crystal violet-iodine crystals form in cell.
◼ Gram-positive
 Alcohol dehydrates peptidoglycan
 CV-I crystals do not leave

◼ Gram-negative
 Alcohol dissolves outer membrane and leaves holes
in peptidoglycan.
 CV-I washes out For further details and
practical application see lab
 Bacteria with No Cell Wall:
Mycoplasmas
◼ Instead, have cell
membrane which
incorporates cholesterol
compounds (sterols),
similar to eukaryotic
cells
This EM shows some typically
pleomorphic mycoplasmas, in this
◼ Cannot be detected by case M. hyorhinis
typical light microscopy
 Acid-fast Cell
Walls

◼ Genus Mycobacterium and Nocardia

◼ mycolic acid (waxy lipid) covers thin
peptidoglycan layer
◼ Do not stain well with Gram stain → use
acid-fast stain
 Damage to Cell Wall
◼ Lysozyme
digests
disaccharide in
peptidoglycan.

◼ Penicillin inhibits
peptide bridges in
peptidoglycan.
Internal Structures: Cell Membrane
Analogous to eukaryotic cell membrane:
◼ Phospholipid bilayer with proteins (Fluid
mosaic model)
◼ Permeability barrier (selectively permeable)
◼ Diffusion, osmosis and transport systems
Different from eukaryotic cell membrane:
◼ Role in Energy transformation (electron
transport chain for ATP production)
Damage to the membrane by alcohols, quaternary
ammonium (detergents), and polymyxin antibiotics
causes leakage of cell contents.
Fig 4.14
Movement of Materials across
Membranes
See Bio 31!

Review on your own if necessary (pages 92 – 94)

Cytoplasm and Internal Structures
Location of most biochemical activities
◼ Nucleoid: nuclear region containing DNA
(up to 3500 genes). Difference between human
and bacterial chromosome?
◼ Plasmids: small, nonessential, circular
DNA (5-100 genes); replicate independently
◼ Ribosomes (70S vs. 80S)
◼ Inclusion bodies: granules containing nutrients,
monomers, Fe compounds (magnetosomes)
Compare to Fig. 4.6
 Endospores
Dormant, tough, non-reproductive structure; →
germination → vegetative cells
Spore forming genera: __________
Resistance to UV and  radiation, desiccation,
lysozyme, temperature, starvation, and chemical
disinfectants
Relationship to disease
Sporulation: Endospore formation
Germination: Return to vegetative state
Sporulation

Fig. 4.21
Green endospores within pink bacilli. Many spores
have already been released from the vegetative cells
 The Eukaryotic Cell
See Bio 31!

Review on your own if necessary (pages 98 – 106)

Eukaryotes compared to prokaryotes:
1. DNA found in nucleus as multiple chromosomes
separated from the cytoplasm by nuclear
membrane.
2. DNA associated with histones.
3. They have organelles e.g., chloroplast and
mitochondria.
4. Cell walls are chemically simple.
5. Cell division by mitosis.

When the glycocalyx helps cells in a biofilm formation

then it is called extracellular polymeric substance (EPS).
Functions for EPS: protects bacteria against dehydration
and sometimes is used as a source of nutrients (sugars) in
low energy environments.

Flagella:
A. Bacteria lacking flagella are called (atrichous)
Flagella arrangements:
B. Peritrichous (distributed over the entire cell)
C. Polar (at one or both poles):
C-1. Monotrichous (single flagellum at one pole)
C-2. Lophotrichous (tuft of flagella at one pole)
C-3. Amphitrichous (flagella at both poles of the cell)
The protein of flagella is called flagellin (globular
protein).
Flagellum has three basic parts:
1. Filament: the long outermost region (made of
flagellin which is arranged in helical structure around
a hollow core.
2. Hook: made of a different protein.
3. Basal body: anchors the flagellum to the cell wall and
plasma membrane (composed of central rod
structure inserted into series of rings).
In gram negative bacteria (two pairs of rings)
1. Outer pair of rings anchored to cell wall
(peptidoglycan and outer membrane).
2. Inner pair of rings anchored to cytoplasmic
membrane.
In gram positive bacteria (one pair of rings)
Anchored to cell wall and cytoplasmic membrane.
Prokaryotic flagellum rotation is either clockwise or
counter clockwise, while for Eukaryotes it’s a wavelike
motion
Flagella motility:
1. Run “swim” and tumbles
2. Swarm across a solid culture medium

The flagellar protein H antigen is used to distinguish

different serovars. e.g., There are 50 different H antigen
for E. coli (O157:H7)

Axial filaments (endoflagella): in spirochetes (spiral

movement
Fimbriae and pili: Mostly found in Gram -ve bacteria
Pili used for motility and DNA transfer: twitching motility
Pseudomonas aeruginosa and E. coli + gliding in
myxobacteria (used for environments with low water)
Protein making the pili is called pilin
Main differences between gram positive and gram-
negative cell walls:
For gram positive cell wall main features:
1. Thick peptidoglycan, containing two types of teichoic
acids: wall teichoic acid and lipotheichoic acid, both
are negatively charged due to their phosphoate
groups. Teichoic acids are composed of: phosphate
group + alcohol (glycerol or ribitol)
2. The periplasmic space consists of only one larger or
its absent (G+ve bacteria)

In gram negative bacteria cell wall main features:

1. Thin peptidoglycan (or murein) layer.
2. No teichoic acids
The periplasmic space maintains the murein layer inside
its structure. The periplasmic space function: contains
degradative enzymes & transport proteins.
3. Cell walls of gram negative bacteria have outer
membranes, which consists of four main structures:
a. Lipopolysaccharides (LPS): composed of Lipid A,
core polysaccharides, O-polysaccharides (O
antigen)
 b. Lipoproteins (LPP): helps in supporting the
structure of the outer membrane.
c. Phospholipids: adds to the negative charge of
cell wall in G-ve bacteria
d. Porins: allow passage of nucleotides,
disaccharides, peptide, amino acids and vitamin
B12. But will restrict passage of antibiotics e.g.,
penicillin
Cell walls and Gram stain:
Some Gram positive bacteria will stain as Gram negative
bacteria (these are called Gram variable e.g., when
cultures of Bacillus & Clostridium ages)
Damage to cell wall:
In Gram positive (complete removal of C.W) forms
protoplast
In Gram negative (incomplete removal of C.W produces
spheroplast), this is due to presence of outer membrane.
In Proteus they swell and become irregularly shape cells
(called L forms Named after Lister Institute)
Cytoplasmic membrane proteins:
1. Enzymatic reactions for energy production:
A. Cellular respiration (ATP production)
B. Photosynthesis (carbohydrate production)
2. DNA replication and cell division
Ribosomes:
In Prokaryotic cells: 70S (30S and 50S)
The 30S is target for streptomycin and gentamicin,
while the 50S is target for erythromycin and
chloramphenicol
In Eukaryotic: 80S (40S and 60S)
The size of the ribosomes in prokaryotic cells is smaller
than their counterparts in Eukaryotic cells.
Endospores: Found only in Gram’s positive bacteria
(Clostridium and Bacillus). The only few exceptions,
one Gram negative bacteria Coxiella burnetii forms
endospore-like structures.
Endospores are used in classification:
1. Terminal
2. Subterminal
3. Central
4. Lateral
 Ch 5
Microbial
Metabolism
 Objectives:
Differentiate between, anabolism, and catabolism.
Identify the components of an enzyme and describe the
mechanism of enzymatic action.
List the factors that influence enzymatic activity.
Explain what is meant by oxidation–reduction.
Describe the chemical reactions of glycolysis.
Explain the products of the Krebs cycle.
Describe the chemiosmotic model for ATP generation.
Compare and contrast aerobic and anaerobic respiration.
Describe the chemical reactions and some products of
fermentation.
Categorize the various nutritional patterns among organisms
according to energy and carbon source.
Catabolic and Anabolic Reactions
• Metabolism: The sum of all chemical
reactions in an organism
• Catabolism: Provides energy and building
blocks for anabolism.
• Anabolism: Uses energy and building blocks
to build large molecules
 Role of ATP in Coupling Reactions
A metabolic pathway is a sequence of enzymatically
catalyzed chemical reactions in a cell.
Metabolic pathways are determined by enzymes, which are
encoded by genes.

Fig 5.1
 Collision Theory
• states that chemical reactions can occur when
atoms, ions, and molecules collide
• Activation energy is needed to disrupt
electronic configurations
• Reaction rate is the frequency of collisions
with enough energy to bring about a reaction.
• Reaction rate can be increased by enzymes or
by increasing temperature or pressure
Enzymes lower
Activation Energy

Compare
to Fig 5.2
 Fig 5.3

Enzymes

• Biological catalysts; specific; not used up in that

reaction
• Enzyme components:
– Apoenzymes, Cofactors, Holoenzymes
– Coenzymes (NAD+, NADP+, FAD)
• Naming of enzymes (see Table 5.1): Lactate
dehydrogenase; Cytochrome oxidase; ligase,
transferase etc.
Mechanism of Enzymatic
Reactions

Compare to
Fig 5.4
 Factors Influencing Enzyme Activity

Enzymes can be denatured by temperature and pH

Fig 5.5c
Fig 5.6

Substrate concentration
influencing enzyme activity
Inhibitors
Competitive Noncompetitive –
vs
inhibitors allosteric inhibitors

Fig 5.7
Sulfa drugs
 Sulfa drugs
• PABA (para-aminobenzoic acid) is used by bacterial
cells to produce folic acid
 Feedback Inhibition

Also known as end-

product inhibition
Controls amount of
substance produced by
a cell
Mechanism is allosteric
inhibition
Fig 5.8
 Energy Production:
Oxidation-Reduction Reactions
• Oxidation = removal of e - Redox reaction =
oxidation reaction paired
-
• Reduction = gain of e with reduction reaction.

Fig 5.9
 Oxidation-Reduction cont.
In biological systems, the electrons are often
associated with hydrogen atoms.
Biological oxidations are often dehydrogenations.

Fig 5.10
The Generation of Phosphorylation:
ATP 1. Substrate level
phosphorylation: transfer
of a high-energy PO4– to
ADP.
2. Oxidative
phosphorylation: transfer
of electrons from one
compound to another is
used to generate ATP by
chemiosmosis.
3. Photophosphorylation
Metabolic Pathways of Energy Production: COH
Catabolism
• Cellular respiration
– Aerobic respiration
– Anaerobic respiration
• Fermentation

The three steps of aerobic respiration

1. Glycolysis (oxidation of glucose to 2 pyruvic acids)
2. Krebs cycle (oxidation of 2 acetyl CoA to 4 CO2)
3. Oxidative phosphorylation (e- transport chain)
Glycolysis
Multi – step breakdown of glucose into pyruvate

Generates
• small amount of ATP (how many?)
• small amount of reducing power – (?)
• Alternative pathways: Pentose phosphate and
Entner-Doudoroff
The Steps of
Glycolysis

Compare to
Fig. 5.12
 Glycolysis & alternative pathways
• Glycolysis or Embden-Meyerhof pathway: the outputs of
glycolysis: 2 pyruvates, 2 NADH, 4 ATPs and 2 net ATP gain
• The pentose phosphate pathway: works by breakdown of
five carbon sugars (pentoses) and it breaks glucose as well.
The intermediates produced by this pathway: nucleic acids,
glucose, amino acids. The output form glucose oxidation:
only one ATP and NADH
• The Enther-Doudoroff pathway (this pathway is found in
gram negative bacteria only): from glucose oxidation the
following are produced: 2 NADPH, one ATP
 Krebs Cycle
• Other names?
• Transition step generates
acetyl-CoA from pyruvate (decarboxylation)
• Acetyl group of acetyl-
CoA enters TCA cycle (tricarboxylic acid cycle)
• Generates ATP and reducing power
• Generates precursor metabolites
 Krebs Cycle

Compare to
Fig 5.13
 Electron Transport Chain
• Formed by series of electron carriers (cytochromes)
located in c.m
• Oxidation/Reduction reactions. Electron carriers (reducing
power) from glycolysis and TCA cycle transfer their electrons
to the electron transport chain
• Generates proton gradient or proton motive force (pmf)

• In chemiosmosis, pmf generates energy via oxidative

phosphorylation
Electron Transport and the Chemiosmotic Generation of ATP

Fig. 5.16
Overview of Respiration and Fermentation

Foundation
Figure
Fig 5.11
Fig 5.17
 Anaerobic Respiration

• Inorganic molecule is final electron acceptor,

e.g.:
-
– NO3
– SO42-

• ATP yield lower than in aerobic respiration

because only part of Krebs cycle operates
under anaerobic conditions.
 Fermentation
• Any spoilage of food by microorganisms (general use)
• Any process that produces alcoholic beverages or acidic dairy
products (general use)
• Any large-scale microbial process occurring with or without
air (common definition used in industry)
Scientific definition:
• Uses an organic molecule as the final electron acceptor
• Does not use the Krebs cycle or ETC
• Energy yield low
• Diversity of end products: _____________________
(see Table 5.4)
The Relationship of
Fermentation to
Glycolysis

Not in book
Also view Fig 5.18
 Location of Carbohydrate Catabolism

Pathway Eukaryote Prokaryote

Glycolysis Cytoplasm Cytoplasm

Intermediate step Matrix of mitochondria Cytoplasm

Krebs cycle Matrix of mitochondria Cytoplasm

ETC Cristae of mitochondria inner membrane

 Energy produced from complete oxidation
of one glucose molecule using aerobic
respiration

Pathway ATP Produced NADH FADH2

Produced Produced
Glycolysis 2 2 0

Intermediate step 0 2 0

Krebs cycle 2 6 2

Total ATP 4 30 4
produced
 ATP produced from complete oxidation of
one glucose using aerobic respiration

By Oxidative Phosphorylation
Pathway By Substrate-Level
Phosphorylation
From NADH From FADH

Glycolysis 2 2x3=6 0

Intermediate step 0 2x3=6 0

Krebs cycle 2 6 x 3 = 18 2x2=4

Total ATP produced 4 30 4

 Carbohydrate Catabolism
• 36 ATPs are produced in eukaryotes

By Oxidative Phosphorylation
Pathway By Substrate-Level
Phosphorylation
From NADH From FADH

Glycolysis 2 6 0

Intermediate step 0 6

Krebs cycle 2 18 4

Total 4 30 4
 Catabolism of Other Compounds

• Polysaccharides and disaccharides

–Amylases for digestion of ___________ (very
common)
–Cellulase for digestion of cellulose (only
bacteria and fungi have this enzyme)
–Disaccharidases

• Lipid catabolism not covered

 Protein Catabolism
Extracellular proteases
Protein Amino acids
Deamination, decarboxylation, dehydrogenation,
desulfurylation
Organic acid Krebs cycle

Decarboxylation
 Biochemical Tests and Bacterial Identification:
Fermentation Tests
Different species produce different enzymes 
test detects enzyme
Mannitol Fermentation:
 Metabolic Diversity among Organisms

• Energy source: Phototrophs vs. Chemotrophs

• Principal carbon source: Autotrophs vs.
Heterotrophs
• Chemoheterotrophs use same organic
compound as energy source and carbon source.
Most medically important bacteria.
• Saprophytes vs. parasites
 Anabolic Pathways
Biosynthesis not covered,
except for
Protein biosynthesis (see Ch 8)
 Ch 6
Microbial
Growth
Objectives:
 Classify microbes into five groups on the basis of preferred temperature
range.
 Explain the importance of osmotic pressure to microbial growth.
 Provide a use for each of the four elements (C, N, S, P) needed in large
amounts for microbial growth.
 Explain how microbes are classified on the basis of O2 needs.
 Identify ways in which aerobes avoid damage by toxic forms of O2.
 Describe the formation of biofilms and their potential for causing infection.
 Distinguish between chemically defined and complex media.
 Justify the use of each of the following: anaerobic techniques, living
host cells, candle jars, selective, differential, and enrichment media.
 Define colony and CFUs and describe how pure cultures can be isolated with
streak plates.
 Explain how microbes are preserved by deep-freezing and lyophilization.
 Distinguish between binary fission and budding.
 Define generation time and explain the bacterial growth curve.
 Review some direct and indirect methods of measuring bacterial cell growth.
Microbial Growth
Microbial growth: Increase in cell number, not cell size!

Physical Requirements for Growth: Temperature

 Minimum growth temperature
 Optimum growth temperature
 Maximum growth temperature
Five groups based on optimum growth temperature
1. Psychrophiles
2. Psychrotrophs
3. Mesophiles
4. Thermophiles
5. Hyperthermophiles

Fig. 6 .3
Fig 6.3: Effect of amount of food on its cooling rate
 Physical Requirements for Growth:
pH and Osmotic Pressure
Most bacteria grow best between pH 6.5 and 7.5:
Neutrophils
Some bacteria are very tolerant of acidity or thrive
in it: Acidophiles (preferred pH range 1 to 5)
Molds and yeasts grow best between pH 5 and 6
Hypertonic environments (increased salt or sugar)
cause plasmolysis
Obligate halophiles vs.
facultative halophiles Fig 6.4
Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings
 Chemical Requirements for Growth: Carbon,
N, S, P, etc.
 Carbon
  Half of dry weight
 Chemoheterotrophs use organic carbon sources

 Nitrogen, Sulfur, Phosphorus

 Needed for ? Vit B1
 Found in amino acids and proteins
(most bacteria decompose proteins)
 S in thiamine and biotin
Vit B7
 Phosphate ions (PO43–)

 Also needed K, Mg, Ca, trace elements (as cofactors),

and organic growth factors
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Chemical Requirements for Growth: Oxygen

O2 requirements vary greatly

Table 6.1: The Effects of Oxygen on the Growth of Various Types of Bacteria

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Toxic Forms of Oxygen
 Singlet oxygen: O2 boosted to a higher-energy state
 Superoxide free radicals: O2–

 Peroxide anion: O22–

 Hydroxyl radical (OH)

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 Fig 6.5
Biofilms
Microbial communities form
slime or hydrogels
Starts via attachment of
planctonic bacterium to surface structure.
Bacteria communicate by chemicals via quorum sensing
Sheltered from harmful factors (disinfectants etc.)
Cause of most nosocomial infections
Clinical Focus: Delayed Bloodstream Infection Following
Catheterization

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings

Culture Media
 Culture medium: Nutrients prepared for microbial growth
 Have to be sterile (not contain living microbes)
 Inoculum: Microbes introduced into medium
 Culture: Microbes growing in/on culture medium
 Chemically defined media: Exact chemical compo-
sition is known (for research purposes only)
 Complex media: Extracts and digests of yeasts,
meat, or plants, e.g.:
 Nutrient broth
 Nutrient agar
 Blood agar
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 Agar
 Complex polysaccharide
 Used as solidifying agent for
culture media in Petri plates,
slants, and deeps
 Generally not metabo-
lized by microbes
 Liquefies at 100°C
 Solidifies ~40°C

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Anaerobic Culture Methods
 Use reducing media, containing chemicals (e.g.:
thioglycollate) that combine with O2

 Are heated shortly before use to drive off O2

 Use anaerobic jar

 Novel method in clinical labs:

Add oxyrase to growth media
 OxyPlate (no need for anaerobic jar)

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Fig 6.5
Capnophiles: Aerobic Bacteria Requiring High
CO2
 Low oxygen, high CO2
Candle jar
conditions resemble those Fig 6.7
found in
 intestinal tract
 respiratory tract and
 other body tissues where
pathogens grow
 E.g: Campylobacter jejuni
 Use candle jar, CO2-
generator packets, or CO2
incubators
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 Selective Media and Differential Media
Selective medium:
Additives suppress
unwanted and encourage
desired microbes – e.g.
EMB, mannitol salt agar etc.

Differential medium:
changed in recognizable
manner by some bacteria 
Make it easy to distinguish
colonies of different
microbes – e.g.  and 
hemolysis on blood agar;
MacConkey agar, EMB,
mannitol salt agar etc.
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Enrichment Media/Culture
 Encourages growth of desired microbe
 Example: Assume soil sample contains a few phenol-
degrading bacteria and thousands of other bacteria
 Inoculate phenol-containing culture medium with the
soil and incubate
 Transfer 1 ml to another flask of the phenol medium
and incubate
 Transfer 1 ml to another flask of the phenol medium
and incubate
 Only phenol-metabolizing bacteria will be growing
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 Pure Cultures
Contain only one species or strain.
Most patient specimens and
environmental samples contain
several different kinds of bacteria
Streak-plate method is commonly used
Colony formation: A population of cells arising from
a single cell or spore or from a group of attached
cells (also referred to as CFU).
Only ~1% of all bacteria can be successfully cultured
Aseptic technique critical!
Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings
 Streak Plate Method

3 or 4
quadrant
methods
Preserving Bacterial Cultures

 Deep-freezing: Rapid cooling of pure culture in

suspension liquid to –50°to –95°C. Good for several
years.
 Lyophilization (freeze-drying): Frozen (–54° to –72°C)
and dehydrated in a vacuum. Good for many years.

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The Growth of Bacterial Cultures
Binary fission – exponential growth

Budding

Generation time – time required for cell to divide

(also known as doubling time)
Ranges from 20 min (E. coli) to > 24h (M. tuberculosis)

Consider reproductive potential of E. coli

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings

Fig 6.13

Figure 6.12b
 Bacterial Growth Curve
Foundation
Illustrates the dynamics of growth Fig 6.15
Phases of growth
 Lag phase
 Exponential or
logarithmic (log) phase
 Stationary phase
 Death phase (decline phase)
Compare growth in liquid and on solid media
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Bacterial Growth Curve: Arithmetic vs.
Exponential Plotting

Fig 6.14
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 Direct Measurements of Microbial Growth

Viable cell counts: Plate counts: Serial

dilutions put on plates CFUs form colonies

Fig 6.16
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Fig 6.17

Figure 6.15, step 1

Additional Direct Measurements
1. Filtration method of choice for low counts
2. Direct microscopic count: Counting chambers
(slides) for microscope

Fig 6.20

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 Estimating Bacterial Numbers by Indirect
Methods

Spectrophotometry to measure turbidity

OD is function of cell number

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Measuring Microbial Growth - Overview

Direct Methods Indirect Methods

 Plate counts  Turbidity
 Filtration  Metabolic activity
 MPN  Dry weight
 Direct microscopic count

Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings

 Chapter 7

The Control
of Microbial
Growth
SLOs
 Define sterilization, disinfection, antisepsis, sanitization, biocide, germicide,
bacteriostasis, and asepsis.
 Describe the microbial death curve.
 Describe the effects of microbial control agents on cellular structures.
 Compare effectiveness of moist heat (autoclaving, pasteurization) vs .dry
heat.
 Describe how filtration, low temperature, high pressure, desiccation, and
osmotic pressure suppress microbial growth.
 Explain how radiation kills cells.
 List the factors related to effective disinfection.
 Interpret results of use-dilution tests and the disk-diffusion method.
 Identify some methods of action and preferred uses of chemical disinfectant
 Differentiate between halogens used as antiseptics and as disinfectants.
 Identify the appropriate uses for surface-active agents.
 List the advantages of glutaraldehyde over other chemical disinfectants.
 Identify the method of sterilizing plastic labware.
 Explain how microbial control is affected by the type of microbe.
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Terminology
 Sepsis: microbial
contamination.
 Asepsis: absence of significant contamination.
 Aseptic surgery techniques prevent microbial
contamination of wounds.
 Antimicrobial chemicals, expected to destroy
pathogens but not to achieve sterilization
 Disinfectant: used on objects
 Antiseptic: used on living tissue
 Nosocomial
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 . . . More Terminology
 Sterilization: Removal of all
microbial life (heat, filtration)
 For food: Commercial sterilization to kill
C. botulinum endospores
 Sanitization: reduces microbial numbers to safe
levels (e.g.: eating utensils)
 Bacteriostatic: Inhibits bacterial
reproduction
 Bactericidal: Kills bacteria
 Fungicide, sporicide, germicide,
biocide
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 Rate of Microbial Death
Bacterial populations subjected to heat or
antimicrobial chemicals die at a constant rate.

Microbial Death
Curve, plotted
logarithmically,
shows this
constant death
rate as a straight Rate: 90% / min
line.
Figure 7.1a
Copyright © 2010 Pearson Education, Inc. Fig 7.1
 How is it possible that a solution containing a million
bacteria would take longer to sterilize than one
containing a half-million bacteria? 7-2

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 Effectiveness of Antimicrobial Treatment
Depends on

 Time it takes to kill a microbial population is

proportional to number of microbes.
 Microbial species and life cycle phases (e.g.:
endospores) have different susceptibilities to
physical and chemical controls.
 Organic matter may interfere with heat treatments
and chemical control agents.
 Exposure time: Longer exposure to lower heat
produces same effect as shorter time at higher
heat.
Copyright © 2010 Pearson Education, Inc.
Actions of Microbial Control Agents

 Alternation of membrane permeability

 Damage to proteins
 Damage to nucleic acids

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 Physical Methods of Microbial Control
 Heat is very effective (fast and cheap).
 Thermal death point (TDP): Lowest temperature at
which all cells in a culture are killed in 10 min.
 Thermal death time (TDT): Time to kill all cells in a
culture
 Decimal Reduction Time (DRT):
Minutes to kill 90%
of a population at a
given temperature

Table 7.2
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 Moist Heat Sterilization
 Denatures proteins
 Autoclave: Steam under pressure
 Most dependable sterilization method
 Steam must directly contact material to be sterilized.
 Pressurized steam reaches
higher temperatures.
 Normal autoclave conditions:
121.5C for 15 min.
 Prion destruction:
132C for 4.5 hours
 Limitations of the autoclave
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 Pasteurization
 Significant number reduction (esp. spoilage and
pathogenic organisms)  does not sterilize!
 Historical goal: destruction of M. tuberculosis
 Classic holding method: 63C for 30 min
 Flash pasteurization (HTST): 72C for 15 sec.
Most common in US.
Thermoduric organisms survive
 Ultra High Temperature (UHT):
140C for < 1 sec.
Technically not pasteurization because it
sterilizes.

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Dry heat sterilization kills by oxidation
 Flaming of loop
 Incineration of carcasses
 Anthrax
 Foot and mouth disease
 Bird flu

 Hot-air sterilization

Hot-air Autoclave
Equivalent 121˚C, 15
170˚C, 2 hr
treatments min
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 Filtration
 Air filtration using high efficiency particulate
air (HEPA) filters. Effective to 0.3 m
 Membrane filters for fluids.
 Pore size for bacteria: 0.2 – 0.4 m
 Pore size for viruses: 0.01 m

Fig 7.4

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 Low Temperature
 Slows enzymatic reactions  inhibits microbial growth
 Freezing forms ice crystals that damage microbial cells
 Refrigeration (watch out for _________________!, deep
freezing, lyophilization

Various Other Methods

 High pressure in liquids denatures bacterial proteins

and preserves flavor
 Desiccation prevents metabolism
 Osmotic pressure causes plasmolysis
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 Ionizing Radiation
 X-rays, -rays, electron beams 
dislodge e- from atoms  production
of free radicals and other highly reactive
molecules
 Commonly used Cobalt-60 radioisotope
 Salmonella and Pseudomonas are particularly
sensitive
 Sterilization of heat sensitive materials: drugs,
vitamins, herbs, suture material
 Also used as ―cold pasteurization‖ of food 
Consumer fears!?
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Nonionizing Radiation: UV light

 Most effective wave legnth

~ 260 nm
 Effect: thymine dimers
 Actively dividing organisms are more sensitive
because thymine dimers cause . . . .?
 Used to limit air and surface contamination. Use at
close range to directly exposed microorganisms
 E.g.: germicidal lamps in OR, cafeteria, and our lab ??

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 Nonionizing Radiation: Microwave
 Wavelength: 1 mm – 1m
 H2O quickly absorbs energy 
release as heat to environment
 Indirect killing of bacteria through heat
 Solid food heats unevenly, why?

Fig 7.5

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 Chemical Methods of Microbial Control
 Few chemical agents achieve sterility.
 Consider presence of organic matter, degree of contact
with microorganisms, and temperature
 Disinfectants regulated by EPA
Antiseptics regulated by FDA
 Use-dilution test
1. Metal rings dipped in test bacteria are dried.
2. Dried cultures of S. choleraesuis, S. aureus, and P.
aeruginosa are placed in disinfectant for 10 min at
20C.
3. Rings are transferred to culture media to determine
whether bacteria survived treatment.

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 Disk-diffusion Method
Disk of filter paper is soaked with a chemical and
placed on an inoculated agar plate; a zone of
inhibition indicates effectiveness.

Fig 7.6

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 Types of Disinfectants
 Phenol = carbolic acid
(historic importance)
 Phenolics: Cresols (Lysol)
- disinfectant
 Bisphenols
 Hexachlorophene
(pHisoHex, prescription),
hospitals, surgeries, Fig 7.7
nurseries
 Triclosan (toothpaste,
antibacerial soaps, etc.)

Phenol and derivatives disrupt plasma membranes

(lipids!) and lipid rich cell walls (??)
Remain active in presence of organic compoundsP
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 Halogens
Chlorine
 Oxidizing agent
 Widely used as disinfectant
 Forms bleach (hypochlorous acid) when added to water.
 Broad spectrum, not sporicidal (pools, drinking water)

Iodine
More reactive, more germicidal. Alters protein synthesis and
membranes.
Tincture of iodine (solution with alcohol)  wound
antiseptic
Iodophors combined with an organic molecule  iodine
detergent complex (e.g. Betadine®). Occasional skin
sensitivity, partially inactivated by organic debris, poor
sporicidal activity.
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 Alcohols

 Ethyl (60 – 80% solutions)

and isopropyl alcohol
 Denature proteins, dissolve
lipids
 No activity against spores
and poorly effective against
viruses and fungi
 Easily inactivated by
organic debris
 Also used in hand sanitizers
and cosmetics
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Table 7.6
Heavy Metals

Oligodynamic action: toxic effect due to metal ions

combining with sulfhydryl (—SH) and other groups
 proteins are denatured.
 Mercury (HgCl2, Greeks & Romans
for skin lesions); Thimerosal
 Copper against chlorophyll containing organisms
 Algicides
 Silver (AgNO3): Antiseptic for eyes of newborns
 Zinc (ZnCl2) in mouthwashes, ZnO in antifungal in
paint
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 Surface Acting Ingredients / Surfactants
 Soaps and Detergents
 Major purpose of soap: Mechanical removal and use as
wetting agent
 Definition of detergents
 Acidic-Anionic detergents Anion reacts with plasma
membrane. Nontoxic, non-corrosive, and fast acting.
Laundry soap, dairy industry.
 Cationic detergents  Quarternary ammonium
compounds (Quats). Strongly bactericidal against against
wide range, but esp. Gram+ bacteria

Soap Degerming
Acid-anionic detergents Sanitizing
Quarternary ammonium compounds Strongly bactericidal, denature
(cationic detergents) proteins, disrupt plasma membrane
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 Chemical Food Preservatives
Sulfur dioxide
wine
Organic acids
Inhibit metabolism
Sorbic acid, benzoic acid, and calcium propionate
Control molds and bacteria in foods and cosmetics
Sodium nitrate and nitrite prevents endospore germination.
In meats. Conversion to nitrosamine (carcinogenic)

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 Aldehydes and Chemical Sterilants
Aldehydes (alkylating agents)
 Inactivate proteins by cross-linking
with functional groups
(–NH2, –OH, –COOH, –SH)
 Glutaraldehyde: Sterilant for
delicate surgical instruments
(Kills S. aureus in 5,
M. tuberculosis in 10 min)
 Formaldehyde: Virus inactivation
for vaccines
Chemical Sterilants for heat sensive material
 Denature proteins
 Ethylene oxide
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Plasma
 Luminous gas with free radicals that destroy
microbes
 Use: Tubular instruments, hands, etc.

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 Hydrogen Peroxide: Oxidizing agent
Inactivated by catalase 
Not good for open wounds
Good for inanimate objects; packaging for
food industry (containers etc.)
3% solution (higher conc. available)
Esp. effective against anaerobic
bacteria (e.g.:
Effervescent action, may be useful for
wound cleansing through removal of
tissue debris
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Microbial
Characteristics and
Microbial Control
Fig 7.11

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