Bacteria PDF

CELL WALL
PLASMA MEMBRANE
RIBOSOMES
NUCLEOID
CAPSULE
PILI
FLAGELLA
CYTOPLASM
General Structure
Gram (+) Bacteria
Gram (-) Bacteria
Growth & Nutrition
Lecturer: Susceptibility Testing

Lea D. Ballares, RMT, M. Bio. Ed.
Bacterial Diseases
Slides by LDBallares
BACTERIA Slides by LDBallares
 Average size: 0.2-10.0 um in diameter
 Basic shapes:
BACILLI
COCCI
SPIRAL
 Elongated cocci:
COCCOBACILLI
 Examples:
1. Listeria monocytogenes
2. Haemophilus influenzae

 Unusual shapes
 Star-shaped Stella
 Square Haloarcula
 Genetically, most bacteria are monomorphic (one

shape)
 A few are pleomorphic based on environmental
conditions

 Pairs: diplococci,
diplobacilli
 Packets of four:
tetrads
 Packets of eight:
octads
 Clusters:
staphylococci
 Chains: streptococci,
streptobacilli
 Pallisade arrangement: Corynebacterium diphtheria

MEDICALLY MEDICALLY MEDICALLY
IMPORTANT IMPORTANT IMPORTANT
COCCI BACILLI CURVED/SPIRAL
Enterococcus spp. Enterobacter spp. CURVED BACILLI
Neisseria spp. Escherichia spp. Vibrio cholera
Streptococcus spp. Klebsiella spp. Campylobacter
Proteus spp. spp.(gull-wing)
Salmonella spp.
Shigella spp. SPIROCHETE
Pseudomonas Treponema pallidum
aeruginosa Borrelia spp. (Lyme
Bacillus spp. disease & relapsing
Clostridium spp. fever)

 Some bacteria may lose their
characteristic shape because
of adverse growth conditions
 CELL WALL DEFICIENT
BACTERIA – shapeless but
revert back to their original
shape when placed under
favorable growth conditions.

PLEOMORPHIC
 No cell wall
 Has the ability to
exist in variety of
shapes
 Example:
Mycoplasma spp.

LIVING STATE WET MOUNT
 To observe shape &
arrangement of
organism
 A drop of bacterial
suspension on slide,
cover it with coverslip
& focus.
LIVING STATE HANGING DROP
 To observe organism’s
motility.
 A hanging drop slide w/
concavity at the center
is used.

FIXED STATE  Adhere organism on
slide & apply stain.

 Bacteria are colorless, transparent &
difficult to see.
It is the process
coloring the
microorganisms with a
dye that emphasizes
certain structures.

PREPARATION  A thin film of a
FOR STAINING solution of microbes
on a slide is a smear.
 A smear is usually
FIXED to attach the
microbes to the slide
and to kill the
microbes.

• Stains consist of a positive and negative ion.
• In a basic dye,
the chromophore
is a cation.
• In an acidic dye,
the chromophore
is an anion.

SIMPLE  It is made up of an aqueous
STAINS solution
 To observe bacterial shape &
arrangement.
 A mordant may be used to
hold the stain or coat the
specimen to enlarge it.

DIFFERENTIAL GRAM STAIN
STAINS  Developed in 1884 by Danish
Bacteriologist Hans Christian
Gram.
 It is one of the most useful
procedures because it divides
the bacteria into 2 large
groups: gram (+) and gram (-).

STEPS Color of Color of
Gram + cells Gram – cells
Primary stain: Purple Purple
Crystal violet
Mordant: Purple Purple
Iodine
Decolorizing agent: Purple Colorless
Alcohol-acetone
Counterstain: Purple Red
Safranin

DIFFERENTIAL ACID-FAST STAIN
STAINS  Acid Fast stain binds only to
those bacteria that have a
waxy material in their cell
walls.
 It is used to identify
Mycobacterium spp.
1. Mycobacterium tuberculosis
2. Mycobacterium leprae
*** Nocardia spp.
DIFFERENTIAL  Cells that retain a basic stain
STAINS (bacteria that have a waxy
material in their cell walls) in
the presence of acid-alcohol
are called acid-fast bacteria
(stained red).
 Non–acid-fast cells (stained
blue) lose the basic stain
when rinsed with acid-
alcohol, and are usually
counterstained to see them.
SPECIAL  Negative staining is the
STAINS process in which the
 Are used to
background & not the organism
color and is stained.
isolate
specific
parts of
microorgani
sms.

SPECIAL  Heat is required to drive a
STAINS stain into endospores.
 Are used to
color and
isolate
specific
parts of
microorgani
sms.

SPECIAL  Flagella staining requires a
STAINS mordant to make the flagella
 Are used to wide enough to see.
color and
isolate
specific
parts of
microorgani
sms.

CULTURE MEDIA – anything that possess
nutritional & environmental requirements for
bacterial growth.

3 TYPES OF CULTURE:
 Pure Culture – made up of one specie of
bacteria
 Mix Culture – made up of organisms
belonging to different species.
 Stock Culture – pure culture of
microorganisms as a source of supply in the
industry.

CLASSIFICATION OF
CULTURE MEDIA
(According to Physical
State):
 Liquid culture
medium – contains no
agar, hardening or
solidifying substances
 AGAR – polysaccharide extracts of seaweed &

commonly used base medium.
CLASSIFICATION OF
CULTURE MEDIA
(According to Physical
State):
 Semi-solid medium –
contains gelatin or 0.5-
1% agar.
 Solid culture medium
– contains 2-3% agar.

CLASSIFICATION OF CULTURE MEDIA
(According to Composition):
 Synthetic culture medium – exact
composition is known
 Non-synthetic culture medium – exact
composition is not known.
 Tissue culture medium – made up of living
cells.

(According to how it is distributed):
 PLATED MEDIUM– distributed in petri dishes

(According to how it is distributed):
 TUBED MEDIUM– distributed in test tubes

(According to use):
SIMPLE  Supports the growth &

MEDIUM multiplication of
microorganisms.
 It is used for routine cultivation
& maintenance of
microorganisms.
 Example: Nutrient Agar

(According to use):
ENRICHMENT  Enhances the propagation of

MEDIUM certain organisms.
 Examples: selenite broth,
tetrathionate broth

(According to use):
ENRICHED  Contains nutritive supplements

MEDIUM needed for the growth of some
organisms.
 Examples: blood agar plate,
chocolate agar plate

(According to use):
DIFFERENTIAL  Distinguishes organisms

MEDIUM growing together by
differences in their cultural
characteristics.
 Examples: EMB, Mc Conkey
Agar

(According to use):
SELECTIVE  Promotes the growth of

MEDIUM desirable organisms but at
the same time inhibiting the
growth of others.
 Examples: Bismuth Sulfide
Agar – isolates Salmonella

Esherichia coli on Eosin Methylene Blue Agar (EMB).
EMB is both a selective & differential media. It is
selective for the growth of gram (-) bacilli

(According to use):
SPECIAL  Specifically prepared to

CULTURE support the growth of specific
MEDIUM organisms.
 Examples:
1. Petragnani medium –
Mycobacterium tuberculosis
2. Thayer Martin – Neisseria
spp.
GENERAL STEPS IN PREPARING CULTURE MEDIA
WEIGH  In plated media: Sterile first
INGREDIENTS before distribution.
DISSOLVE  In tubed media: Distribute
INGREDIENTS first before sterilization.
ADJUST TO
PROPER PH
DISTRIBUTION IN STERILE
STERILIZATION
PETRI DISHES
TECHNIQUES  LIQUID CULTURE MEDIUM –
OF inoculate the organism & shake.
INOCULATION  SLANT TUBED MEDIUM –
streaking from the bottom with a
zigzag fashion
INOCULATION
 BUTT MEDIUM – stabbing
– To introduce
microorganisms  BUTT/SLANT – stab, then
into a culture streak
medium or host.  PLATED MEDIUM - streaking

METHODS OF STREAKING PLATED MEDIUM
 RADIAL STREAK METHOD – place the
inoculum one side of the plate, then bring on
the other side concentric fashion.
 OVERLAP STREAK METHOD – keeps on
overlapping, used in sensitivity testing.
 MULTIPLE STREAK MEDIUM – divide the
medium into several division, then streak
separately

METHODS OF STREAKING PLATED MEDIUM
 INTERRUPTED STREAK METHOD – start
streaking on one side of the plate, stop, then
turn for 180 degrees & streak again
 MULTIPLE INTERRUPTED STREAK
METHOD – used to obtain pure isolated
colonies.

Works well when the bacteria is present in high
numbers.
 A pure culture contains only one species or
strain
 A colony is a population of cells arising from
a single cell or spore or from a group of
attached cells
 A colony is often
called a colony-
forming unit
(CFU)

TYPES OF COLONIES:
COLONY
 S or Smooth colonies –
MORPHOLOGY
with uniform texture &
homogenicity, associated
w/ virulent organisms
 M or Mucoid Colonies –
exhibits slimy & watery
appearance, associated w/
capsulated & virulent
organisms.
TYPES OF COLONIES:
COLONY
 R or Rough Colonies –
MORPHOLOGY
granulated in appearance.

 A colony contains millions
COLONY of organisms.
MORPHOLOGY
 Size, shape, color,
elevation & margin are
observed to identify the
bacteria.

GROWTH PATTERNS IN PLATES
• SIZE: pinpoint, small,

moderate, or large
• PIGMENTATION: color
of colony.
• FORM: The shape of
the colony.
(1) Circular: unbroken
peripheral edge.
(2) Irregular: indented
peripheral edge.
(3) Rhizoid: rootlike
spreading growth.
MARGIN: The
appearance of the outer
edge of the colony .
(1) Entire: sharply defined,
even.
(2) Lobate: marked
indentations.
(3) Undulate: wavy
indentations
(4) Serrate: toothlike
appearance
(5) Filamentous:
threadlike, spreading
edge

• ELEVATION: The degree

to which the colony
growth is raised on the
agar surface.
(1) Flat: elevation not
discernible.
(2) Raised: slightly
elevated.
(3) Convex. Dome
shaped elevation.
(4) Umbonate. Raised
with elevated convex
region

 It refers to the number of cells, not the cell
size.
 “Growing microbes” increases in number.

 Bacteria multiply by a
process called binary
fission.
 The time required for the
cell to divide or the
population to double is
called generation
(doubling) time.

STAGES OF BINARY FISSION
Bacilli following division
Chromosome division, cell growth

by lengthening
Chromosome divided, cell fully lengthened,
growth of envelope,Chromosomes
segregated
Cross wall completed
Daughter cells separate

Rate of reproduction is equivalent
to death rate due to exhaustion of
nutrients
Bacteria are multiplying at a

constant & maximum rate
The rate of reproduction has
completely stopped due to lack of
nutrients & formation of toxic
Bacteria are adjusting to a new products
environment, synthesizing
enzymes & actively metabolizing
 All organisms, whether they be
NUTRITIONAL bacteria, humans, or trees,
REQUIREMENTS
need a constant supply of food
in order to live.
 Water which is absolutely
essential for cellular function.
 Carbon which is the major
structural element in cell
constituents

 Energy required for cellular
NUTRITIONAL growth.
REQUIREMENTS
 Nitrogen which is also an
important structural elements,
being a constituent of proteins
& nucleic acids
 Traces of other elements
required for life processes.

 All organisms, whether they be
NUTRITIONAL bacteria, humans, or trees,
REQUIREMENTS
need a constant supply of food
in order to live.
 FASTIDIOUS – organisms that
has demanding nutritional
requirements.

REQUIREMENTS FOR
GROWTH:
1.Physical –
temperature, pH,
osmotic pressure
2.Chemical – Water,
sources of carbon &
nitrogen, minerals,
oxygen & organic
growth factors.
PHYSICAL TEMPERATURE
REQUIREMENTS
 PSYCHROPHILES – “cold
loving” microbes.
 MESOPHILES –
moderate-temperature
loving microbes
 THERMOPHILES – “heat
loving” microbes.

PHYSICAL TEMPERATURE RANGE
REQUIREMENTS  MINIMUM GROWTH
TEMPERATURE – is the
lowest temperature at
which species will grow
 OPTIMUM GROWTH
TEMPERATURE – is the
temperature at which the
species grows best.

PHYSICAL TEMPERATURE RANGE
REQUIREMENTS  MAXIMUM GROWTH
TEMPERATURE - is the
highest temperature at
which growth is possible.

PHYSICAL TEMPERATURE
REQUIREMENTS
PSYCHROTROPHS
 Grow between 0°C and 20-
30°C
 Cause food spoilage
 Also known as moderate
psychrophiles or facultative
psychrophiles

PHYSICAL pH
REQUIREMENTS
 Most bacteria grow between
pH 6.5 and 7.5.
 Molds and yeasts grow
between pH 5 and 6 (greater
pH range compared to
bacteria).

PHYSICAL pH
REQUIREMENTS
 ACIDOPHILIC BACTERIA
– remarkably tolerant of
acidity
 BASOPHILIC BACTERIA –
grows at pH near neutrality.

PHYSICAL  OSMOTIC PRESSURE is
REQUIREMENTS the pressure that is exerted
on a cell membrane by
OSMOTIC
solutions inside & outside
PRESSURE
the cell.
 What are the effects of the
ff. solutions in a bacterial
cell?
1. Hypertonic
2. Isotonic
3. Hypotonic
PHYSICAL HYPERTONIC SOLUTION
REQUIREMENTS  The cell membrane & cytoplasm
shrink away from the bacterial
OSMOTIC
cell wall – PLASMOLYSIS.
PRESSURE
 Salts & Sugars are added to
certain foods to preserve them.
 Bacteria in hypertonic
environment will die as a result of
desiccation.

PHYSICAL HYPOTONIC SOLUTION
REQUIREMENTS  If a bacterial cell is placed in a
hypotonic solution, the fluid
OSMOTIC
pressure w/in the cell increases
PRESSURE greatly.
 If the pressure becomes so great
& cell bursts, cytoplasm escapes
from the cell – PLASMOPTYSIS.

PHYSICAL ISOTONIC SOLUTION
REQUIREMENTS  In an isotonic environment, water
neither leaves nor enter the cell.
OSMOTIC
PRESSURE

PHYSICAL HALOPHILES
REQUIREMENTS  halo referring to “salt” & philic
OSMOTIC meaning to “love”
PRESSURE  Bacteria that loves salty
environment.
 Example: Vibrio cholera

PHYSICAL HALODURIC
REQUIREMENTS  Organisms that do not prefer
OSMOTIC to live in a salty environment
PRESSURE but are capable of surviving
there.
 Example: Staphylococcus
aureus

PHYSICAL BAROPHILES
REQUIREMENTS  Organisms that thrive deep in
BAROMETRIC ocean & oil wells, where
PRESSURE atmospheric pressure is very
high.

CHEMICAL  Water in the liquid state is
REQUIREMENTS essential for the existence of all
living organisms.
WATER OR  Approximately 75% water is
MOISTURE present in the cells of every living
organism, including bacteria.
 This amount of water is required
to maintain the cell in an active
state, and without liquid water,
living organisms will not be able to
grow or reproduce.

CHEMICAL  Substances such as sugar and
REQUIREMENTS salt makes the water unavailable
for bacteria.
WATER OR  The amount of water available for
MOISTURE microbial growth is referred to as
Water Activity.

CHEMICAL  Oxygen is essential for the
REQUIREMENTS growth of many bacteria, but
OXYGEN for others it is lethal.
 All bacteria have cell
components that are sensitive
to oxygen and metabolic by-
products of oxygen.
 Organisms that live in air
have enzymes that detoxify
these products.
CHEMICAL • Obligate Aerobes absolutely
REQUIREMENTS require oxygen for their
OXYGEN growth.
Example: Mycobacteria spp.
• Obligate Anaerobes are
those that are unable to grow
in the presence of free
oxygen because O2 kills or
inhibits them.

CHEMICAL • Aerotolerant anaerobe does
REQUIREMENTS not require oxygen, grows
better in the absence of
OXYGEN oxygen, but can survive in the
presence of oxygen.
• Facultative anaerobes are
capable of surviving in either
the presence or absence of
oxygen.
Examples: Enterobacteriaceae,
most Streptococci & Staphylococci.
CHEMICAL • Microaerophiles need a
REQUIREMENTS small quantity of oxygen, but
OXYGEN large quantities inhibit their
growth or even kill them.

A. Aerobic
B. Anaerobic
C. Facultative
D. Microaerophilic
E. Aerotolerant
anaerobe

CHEMICAL  An atmosphere of 5-10%
REQUIREMENTS CO2 is required by some
organisms, referred to as
CAPNOPHILES.
 Examples: Neisseria spp.,
Campylobacter spp.,
Haemophilus spp.


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CELL WALL

Gram (+) Bacteria

Gram (-) Bacteria

Growth & Nutrition

Lecturer: Susceptibility Testing

BACTERIA Slides by LDBallares

 Genetically, most bacteria are monomorphic (one

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

 AGAR – polysaccharide extracts of seaweed &

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

SIMPLE  Supports the growth &

BACTERIA Slides by LDBallares

ENRICHMENT  Enhances the propagation of

BACTERIA Slides by LDBallares

ENRICHED  Contains nutritive supplements

BACTERIA Slides by LDBallares

DIFFERENTIAL  Distinguishes organisms

BACTERIA Slides by LDBallares

SELECTIVE  Promotes the growth of

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

SPECIAL  Specifically prepared to

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

• SIZE: pinpoint, small,

BACTERIA Slides by LDBallares

• ELEVATION: The degree

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

Bacilli following division

Chromosome division, cell growth

Daughter cells separate

BACTERIA Slides by LDBallares

Bacteria are multiplying at a

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares

BACTERIA Slides by LDBallares