FUNDAMENTAL OF MICROBIOLOGY
Chapter 3
Types of Microorganism - Bacteria
LEARNING OUTCOMES
• Understand the classification of bacteria
• List the structure, size and shape, and
reproduction properties
• Identify the influencing factors on bacteria
infestation
• Describe 4 stages occur in bacterial growth
curve
Bacteria
Bacteria
Known a prokaryotic cell
Need microscope to see (length about 0.5 – 20
µm)
No membrane bound nucleus
Have ribosomes and cytoplasm, but no other
organelles
Contain DNA stored in nucleoid
DNA present in a long circular molecule
Can be found on most materials and surfaces
1 micron (µ) / micrometre (µm) = 1⁄1000 mm
Bacteria
• Are found everywhere :
– Air
– Water
– Soil
– Plants
– Animals
BACTERIA
Bacterial : Structure
• Cell Wall - outermost
structure of the cell.
• Made up of a substance
called peptidoglycan ( a long
chain of sugars linked to
amino acids ).
• Penicillin destroys bacteria
by interfering with the
peptidoglycan molecules.
Bacterial : Structure
• Cell Membrane - found
beneath the cell wall.
– ATP production (energy),
takes place on the folds of
the membrane.
• Bacteria that have capsules
are said to be encapsulated.
• Most pathogenic bacteria are
encapsulated.
How do bacteria reproduce?
• Grow in number not in size
– Humans grow in size from child to adult
• Make copies of themselves by dividing in half,
known as binary fission. This process leads
the bacteria to reproduce every 20 minutes –
rapid bacterial contamination
Influencing Factors on Bacteria
Infestation
• For their optimal growth, bacteria require:
– Oxygen
– Carbon Dioxide
– Temperature
– Light
– pH
– nutrients
• Bacteria also require a source of nitrogen and a
number of salts to have a supply of potassium,
magnesium, iron, phosphate and sulphate.
Influencing Factors on Bacteria
Infestation
Oxygen:
• The need of oxygen for particular bacterium reflects,
it’s mechanism to meet the requirement of energy.
i) Aerobic bacteria
ii) Anaerobic bacteria
iii) Facultative bacteria
Influencing Factors on Bacteria Infestation
Temperature :
• Bacteria vary in their requirement of temperature for
growth.
• For each species, there is temperature range and
growth does not occur above.
• The temperature at which growth occurs best is
known as the ‘optimum temperature’ which most
pathogenic bacteria are 37 degree.
Influencing Factors on Bacteria
Infestation
Carbon dioxide :
• All bacteria require small amounts of carbon dioxide
for growth.
• This requirement is usually met by the carbon
dioxide present in the atmosphere or produced
endogenously by cellular metabolism.
Influencing Factors on Bacteria
Infestation
Light :
• Bacteria (except the phototrophic species)
grow well in the dark.
• Sensitive to ultraviolet light and other
radiations.
• Cultures die if exposed to sunlight.
Influencing Factors on Bacteria
pH : Infestation
• The pH of medium of growth bacteria has
profound effect upon the multiplication of
organisms.
• Most pathogenic bacteria require a pH of 7.2
– 7.6 for their optimal growth.
• The pH of the growth medium keeps a
changing depend to the reaction performed
by the bacterium with the nutrients
Influencing Factors on Bacteria
Nutrient :
Infestation
• Bacteria need food and water
• Different bacteria have different requirements
for certain substances such as vitamins and
amino acids but all bacteria require carbon
dioxide
• Most grow better in neutral or slightly alkaline
solutions, all need phosphate
Classification of Bacteria
• Shape of bacteria
• Nutrition
• Staining
• Oxygen requirements
Bacteria
• 3 basic shapes :
– Rod shaped called bacilli
• Pairs – diplobacilli
• Chains – streptobacilli
• Very short - coccabacilli
– Round shaped called
cocci
• single cells – monococci
• Pairs – diplococci
• Chains - streptococci
Bacteria
• 3 basic shapes (Cont’) :
– Spiral shaped
(spirochaete)
– Some exist as cluster
shape, eg:
staphylococcus (staphyl
means cluster); (coccus =
spheroidal)
Bacteria
Shape of Bacteria
• Diplococcic
arrangement
Shape of Bacteria
Rod-shaped bacilli
Shape of Bacteria
Bacilli with flagella Streptobacilli
Nutrition
• Bacteria are placed in 2 categories, depending on
how they get their food.
• 2 categories :
– Autotrophic bacteria
- Can make their own food
– Heterotrophic bacteria
- Get food from others
Heterotrophic Bacteria
• Heterotrophs must get their
food from a source of pre-
formed organic matter:
• (A) Saprobes- feed on the
remains of dead plants and
animals.
• (B) Parasites (pathogens) -
live on or in the organism
and cause disease. For
example, Mycobacterium
tuberculosis.
Staining
• Classified bacteria using staining technique
• 2 groups of separated organism:
– Gram positive
– Gram negative
Gram stain
• Bacteria are colorless and invisible with naked
eyes
• First smear the substance to be stained (eg:
sputum, pus, etc) onto a slide,
• Then, heat it to fix the bacteria on the slide
Gram stain (cont’)
Gram stain (cont’)
Gram stain +ve Gram stain -ve
Outer thick peptidoglycan layer Thin peptidoglycan layer (5
(60 – 100%) – 10%)
Outer membrane contain low Outer membrane contain high
lipid content lipid content
No periplasmic space There is a space between the
layers of peptidoglycan and the
cell membrane called periplasmic
space
Survive in dry environments Kill more quickly by drying. Grow
better. Often found on skin & better in the presence of toxin
dust chemical and aqueous
Gram stain (cont’)
Gram stain +ve (examples)
• S. saprophyticus • C. jelkenium
• S. bovis • Lactobacillus sp.
• C. urealyticum • [Link]
• Staph aureus • Veillonella
• B. cereus • Eubacterium
• S. pneumoniae • Clostridium sp.
• A. adiacens • C. difficile
• S. mitis group • C. perfringens
• S. agalactiae • Listeria monocytogenes
• S. lugdunensis • Erysipelothrix rhusiopathiae
• Arcanobacterium bemolyticum
Gram stain (cont’)
Gram stain –ve (examples)
• Escherichia coli • Alkaligenes
• Morganella • Burkholderia cepacia
• N. gonorrhea • H. parainfluenzae
• Moraxella catarrhalis • Pasturella multocida
• N. meningitidis • Kingella kingae
• Klebsiella pneumoniae • Brucella sp.
• Aeromonas hydrophila • C. jejuni
• Providencia • Haemophilus aphrophilus
• Enterobacter cloacae • Stenotrophomon
• Cariobacter hominis
• H. influenzae
Oxygen Requirements
Bacteria can be divided into 3 groups with regard to
their need of oxygen :
A) Aerobes
• Which need oxygen to grow, found on the surface
of wound, on skin and mucous membrane.
Eg : Pseudomonas aeroginosa
B ) Anaerobes
• which cannot grow in the presence of oxygen.
These will particularly affect deep wounds where
tissue is dead.
• Eg : stab, gun shot, crush wounds
• They include the Clostridium sp. Which cause
tetanus and gas gangrene.
Oxygen Requirements
C ) Facultative
• Which can grow with or without oxygen .
• Eg : Staphylococci (gram +ve) and
Escherichia coli (gram –ve)
Bacterial Growth Curve
Bacterial Growth Curve
• Growth Rate = doublings per hour
• In the presence of fresh growth medium, a
bacterium shows the following 4 phases :
1. Lag Phase
• Occurs immediately after inoculation
• Cells do not grow because the bacteria adapt
themselves to new environment
Bacterial Growth Curve
2. Growth Phase (exponential phase)
• Known as Log Phase.
• During this phase the microbe is growing at
the maximum rate possible due to nutrients
present in the medium are utilised by the
bacteria and daughter cells.
• Cells per volume increases dramatically.
• Most research is performed on cells during
this phase.
Bacterial Growth Curve
3. Stationary Phase
• Growth level stagnated once the exhaustion of
nutrients occurs.
• Cells per volume does not increase or
decrease.
• The morphology of bacteria is best developed
in this phase.
• Due to : depletion of nutrients & increase in
waste products.
Bacterial Growth Curve
4. Death Phase / logarithmic phase / decline
phase
• Death rate exceeds growth rate
• Cells per volume decreases
• Due to : very depletion of nutrients and very
high concentrations of waste products
Bacteria Media & Culture
Media
• Nutrients (agar2, pH indicator, proteins and CHO) used to
grow organisms outside of their natural habitats
• Must be prepared in a sterile way prior to being inoculated
with bacterial sample, so that when particular type of
bacteria is cultivated (cultured) on the medium, it is the
only type of bacteria present
Culture
• The propagation of microorganisms using various media
Culture Media
Usage :
• Used to grow bacteria
• Encriched the numbers of bacteria
• Differentiate among different kinds of bacteria
• Determine the sensitivity of antibiotic
Culture and Medium
• Culture is the term given to microorganisms
that are cultivated in the lab for purpose of
identifying and studying them
• Medium is the term given to the combination
of ingredients that will support the growth
and cultivation of microorganisms by all the
essential nutrients required for growth
Culture media
Proper Storage of Culture Media
• The expiry date of each product should be shown on
the labels of culture media.
• Light :
– all prepared culture media and their components
should be stored away from light and avoid
exposure to direct sunlight at all times.
• Humidity :
– Avoid preparing and keeping the culture media in
hot, steamy media preparation room.
– Cold room or adequate storage cupboard are
preferable storage area.
Proper Storage of Culture Media
• Temperature and time:
– Culture media : kept sealed, unopened container
should be stored at room temperature 15-20°C.
Shelf life : 1-5 years.
– Broth media : Do not freeze. Store at 2-8°C. Shelf
life : 6 months – 2 years.
