Introduction to Microbiology

NT20203
Food Microbiology
MICROBIOLOGY – STUDY ON MICROORGANISMS

 Too small to be seen with unaided human eye

 Cell or other structures are simple and less specialized
 Can be cultured in laboratory
 Traditional methods of growing microorganisms – pure
culture techniques
 An organism is isolated from it natural habitat and
inoculated into a sterile medium where it can grow
well
 Handling with aseptic techniques – avoid organism
from contaminate lab environment or other
microorganisms contaminate the pure cultures.
 Bacteriology – study on bacteria
 Mycology – study on fungi
 Parasitology – study on protozoa and parasitic
worms
 Phycology – study on algae
 Virology – study on virus
 Immunology – immune system
 Specialization
 Medical – pathology, immunology and
epidemiology
 Agricultural – livestock, plants
 Industrial – biotechnology, industrial processes
 Food – food spoilage, production and food
preservation
Topics of interest
1. Food spoilage
2. Foodborne diseases
3. Food hygiene
4. Food preservation
5. Quality assurance/control
6. Food fermentation
7. Water quality
8. Laboratory management
 Prokaryotic Vs Eukaryotic
Distinguishing characteristics of Prokaryotic:
 Their DNA (genetic material) is not enclosed within a
membrane
 DNA is not associated with histones (special
chromosomal protein)
 Lack of membrane enclosed organelles
 Cell wall always contains the complex polysaccharides
peptidoglycan.
 Usually divided by binary fission.

eg. All bacteria

 Eukaryotic cells
 DNA is found in Nucleus separated by nuclear
membrane or found in multiple chromosomes
 Consistently associated with chromosomal protein
 Have number of membrane enclosed organelles, eg
mitochondria, endoplasmic reticulum, Golgi, lysosomes,
etc..
 Cell wall are chemically simple (if present)
 Usually divided by mitosis – chromosome replicate into
identical set.

eg. Animals, plants, fungi, protozoa, algae

 Compound Light Microscope

 Components and their function (Table)

 Resolution: resolving power
 Ability to distinguish fine detail and structure - ability to
distinguish between 2 points at a specified distance.
 Wavelength shorter, resolution 
 To achieve high magnification with good resolution -
immersion oil between slide and objective lens
 Reduce lost of light rays after passed through the specimen
 Same refractive index as glass – same effect as increasing
the diameter of objective lens.
 Dark-field
 Phase contrast,
 Fluorescence,
 Transmission electron (TEM),
 Scanning electron (SEM),
 Confocal etc…
Magnification properties
Gram stained bacteria as viewed with light microscope (left) and SEM (right)
 The structure of microorganisms
Bacteria
 Simple, singe-celled organisms, size: 0.2-3 um x 05.-10 um
 Can only be seen by microscopic techniques.
 Bright field microscope – used in lab to observe bacteria and yield
important info for their identification: shape of cell, arrangement,
Gram stain and spores.
 Bacteria cells are normally stained with colour dyes to increase
contrast
 Gram + and -
 For detail structural: electron microscope – resolving power and
higher magnification
 X 1000 (lab microscope) – 0.2 um x 100,000 (TEM/SEM) – 2.5 nm
 Bacteria structure
 All bacteria cell have:
 Cell wall (except mycoplasma)
 Plasma membrane
 Cytoplasm
 Single chromosome (DNA)
 Ribosomes
 Only some bacteria have:
 Capsules
 Glycocalyx
 Fimbriae (pili)
 Flagella
 Spores
 Inclusion granules (storage)
Prokaryotic Cell Structure
 BACTERIAL CELL WALL
 Rigidlayer surround the cell membrane –
giving the cell its shape
 Protect cell from osmotic lysis and mechanical
damage
 Contains
polymer of amino sugars: N-acetyl
glucosamine and N-acetyl Muramic acid.
 Form 3-D structure: peptidoglycan (Murein or
Mucopeptide or Mucocomplex)
 Gram + or – bacteria differ in chemical
structure
 Gram + almost entirely of peptidoglycan, teichoic
acid is present. Covalent linked between teichoic
acid and peptidoglycan increase strength
 Gram – cells have thinner peptidoglycan layer
(1/10), without teichoic acid
 Cell wall (Cont..)
 There is a layer call lipopolysaccharide (outer
membrane/envelope) – very complex and
responsible for antigenicity, induce antibody
formation and toxicity of Gram – bacteria (fever,
shock, other symptoms of diseases)

 Impermeable protective barrier from toxic

chemicals eg. antibiotic will not penetrate the
outer membrane

 Pore proteins – as channel for movement of

nutrients
 Cell membrane
 Thin flexible lipid and protein envelope that defines
the boundary of cytoplasm and external environment
 Phospholipid bilayer with protein molecules (integral
protein) and other proteins embedded.
 Controls movement of materials in and out cell.
Contains permeases (transport enzymes) move
nutrients through the membrane
 Genetic material is bounded to the cell membrane
during division – redistribution of genetic materials to
new cells.
 Extracellular enzymes are secreted at cell surface.
 Bacterial genome
 Single circular chromosome – double
stranded DNA molecule controls cell
development and metabolic activities.
 Plasmid – small circular structure make up
of 5-100 genes carrying information that is
not essential for survival but may helps in
adapt to changing environment. eg antibiotic
resistance or break down of unusual
compounds
 Can be transferred between strain/species
 Capsules
 Surface layers outside cell wall. Also called slime layer if the
layer are easily detached and diffuse into the surroundings.
 Normally made of polysaccharide (CHO as C source) or
polypeptide (amino acids as a source)

Functions:
– Protect cell from dehydration
– Enable bacteria to stick to inert surfaces or stick to each
other
– Protective barriers against antibiotics and ingestion of
phagocytes
– Capsular polysaccharide may act as receptor for
bacteria viruses attachment
– Protect cell against viral (bacteriophage)

 Glycocalyx – enables bacteria to stick to each other and

also surfaces/host cells
• thick peptidoglycan • thin peptidoglycan
• teichoic acid • no teichoic acid
- ion flow • outer membrane
- protection - LPS = endotoxin
- antigen specificity - protection
- porins
Peptidoglycan layer (yellow); protein (purple); teichoic acid
(green); phospholipid ( brown); lipopolysaccharide (orange).
LPS
1. Chromosomal DNA
2. Plasmids
 Escherichia coli

Enterococcus faecium
Binary fission - one cell splits into two cells, offspring are genetically
identical to parent
Bacterial conjugation - a form of sexual reproduction where bacteria
exchange genetic information before dividing, offspring have new genes
Transformation - bacteria incorporate genes from dead bacteria
Transduction - viruses insert new genes into bacterial cells. This method is
used in biotechnology to create bacteria that produce valuable products
such as insulin
Glycocalyx
• Complex exopolysaccharides
• Adhesion
• Protection from
- biocides
- antibiotics
- bacteriophage
- free-living amoebae
- WBC
 Bacterial flagella
 10-20 um in length and 0.1-0.7 in diameter
 Originate from cell membrane and appear to be
attached to membrane and cell wall by basal body.
 Made of unique helical protein – flagellin
 Responsible in the movement of bacteria in liquid
medium.
 Flagella arrangement – peritrichous Vs polar
 Absence or presence and types of flagella – bacteria
classification.
 Bacillaceae and Enterobacteriaceae – peritrichous
 Rapid darting type of movement
 Psudomonadaceae – polar
 Leisurely in which flagella rotation is synchronised
 Bacteria flagella (cont..)
 Cannot be seen with ordinary microscope and
staining of flagella is not easy/not reliable
 Use indirect methods for examination – hanging drop
technique and motility agars
Flagella attachment in bacteria
 BACTERIAL MOVEMENT
 Thebacterial flagellum can rotate both
counterclockwise and clockwise
 Clockwise rotation results in a tumbling motion and
changes the direction of bacterial movement.
Counterclockwise rotation leads to long, straight or
curved runs without a change in direction
 Motility serves to keep bacteria in an optimum
environment via taxis. Taxis is a motile response to an
environmental stimulus
 chemicals (chemotaxis),
 light (phototaxis),
 osmotic pressure (osmotaxis),
 oxygen (aerotaxis),
 temperature (thermotaxis).
Chemotaxis is a response to a chemical gradient of attractant or
repellent molecules in the bacterium's environment. In an environment
that lacks such a gradient, the bacterium moves randomly.
If they are exposed to a repellent gradient, it tumbles less frequently (has
longer runs) as it moves up the gradient, but tumbles at the normal rate if
it travels down the gradient
 BACTERIAL SPORE
 Found in relatively small number of bacterial
species.
 Resistance of spores to adverse
environmental circumstances:
• Heat
• Chemical including disinfectants and preservatives.
• Freezing
• Extreme pH
• Dehydration and freeze drying
• Low aw

 Some processing operations are designed to

kill bacterial endospores, e.g. canning of low
acid foods – C and t to kill Clostridium
botulinum.
Bacterial spores
 Bacterial spores (Bacillus subtilis)
labeled with luminescent

Malachite green stain

 BACTERIA SPORES (CONT..)
• Spores have structural, physiological and
biochemical characteristics that are very
difference from vegetative cells:
• outer spore coat (protein) resist to chemical agent and pH
• highly dehydrated state (15% H2O)
• low level of metabolic activity
• relatively few enzymes – lower molecular wt and heat
resistant

• Bacterial spores difficult to stain – spore wall

resists stain penetration.
• Require drastic treatment – heating stain in contact with
spores or applying chemical such as phenol.
• eg. Malachite + heat for 5 minutes
 BACTERIA SPORES (CONT..)
• A spore remains dormant in environment after
release until stimulated to germinate
• Chemical or physical stimuli
• Heat shock – require heating at high temperature to
induce germination
Important some food poisoning outbreak caused by
spore-forming bacteria.
Heating the food induce dormant spores to germinated
after food has cooled down and form new vegetative
cells – reproduce & grow to cause food poisoning.
• Life cycle of spore-forming bacterium (Figure)
The developmental cycle of the Endospore
Why do we need to study microorganisms?

• Impact on Human Health

• Balance of Nature - food source, play a role in
decomposition, help other animals digest grass (cattle,
sheep, termites).
• Environmental – provide safe drinking water;
development of biodegradable products; use bacteria
to clean up oil spills, etc. – called bioremediation.
• Industrial – foodstuffs (beer, wine, cheese, bread),
antibiotics, insulin, genetic engineering
• Agricultural - research has led to healthier livestock
and disease-free crops.
 Commercially valuable microbial products
• Antibiotics. Over 10,000 different antibiotics have been
isolated and characterized. About 160 are produced in
commercial quantities. Pfizer is one of the major antibiotic
producer.
• Biocides. Example: Bacillus thuringiensis produces crystalline
material that destroys insect guts of lepidopterans
(caterpillars), very effect and specific pesticide.
• Foods produced by fermentation. Includes cheeses,
fermented milk products (yogurt, sour cream, buttermilk, etc.),
leavened breads, saurkraut, pickles, soy sauce, more.
• Beverages produced by fermentation. Includes wine, beer and
ale, fermented grain mashes used to produce all distilled
liquors.
• Foods and flavoring agents. Includes mushrooms, yeast cells,
vinegar, nucleotides, amino acids, vitamins, organic acids.
• Enzymes. Over 500 tons of purified enzymes are produced
annually, enormous commercial value. Include proteases (used in
detergents), amylases (used in production of syrup, brewing,
baking, animal feed, detergents), rennin (used in cheese
making), Taq polymerase (used in PCR).
 Food Laboratory
fermentation management

Food hygiene
Water quality

Major areas of food

microbiology

Food Foodborne
preservation disease

Food Quality control

spoilage

Main areas of interest/knowledge in food microbiology