Chap 2: Structure + Morphology Comparison between bacteria, yeast and mold

Bacteria Fungi (Eucaryotic)

(Procaryotic)
Including archaea Yeast (Eukaryotes) Mold

Morphology Size: 1µm Unicellular structure, immobile Color: some molds are
Shape: Cocci (spherical shape), Bacilli (rod-shape), Spirilium Size: 1 dimension 5-7 µm colorful from spore color
(spiral shape) Shape: sphere, egg shape, oval Hyphae without
Arrangement: shape bottle shape, long tube embranchment
The Cocci shape Shape : mycelium (septate
 Coccus or aseptate)
Arrangement: polymorphism
 Streptococci (long chain) Penicillium colony
 Diplococci (form pairs)
such as pseudomycelium, Single cell (aseptate hypae)
 Sarcina (3 planes, producing cubical packets of eight mycelium or multi cell (septate hypae)
cells ) Colony morphology: Similar Size: 1-10um x(>)10cm
 Tetrad (2 plane to form square groups of four cells) to bacterial colonies, yeast long filament (hyphae).
 Staphylococci (random planes, grapelike cluster) colonies are colorless and Can be very long.
The Bacilli (bacillus and bacterium) typically non-mucoid or dry in Form colony on solid
 Bacillus texture. culture
 Coccobacillus Cellular structure :
 Spore - former multicellular filaments
 Diplobacillus (in twos) Eukaryote
 Palisades (like a stack) Flamentus fungite
 Streptobacillus (in chains) A hyphaee : branching
Colony morphology: same with yeast, maybe color tubular structure, (2-10 µm
in diameter) which is
usually divided into cell-like
units by crosswalls called
septa
Arrangement:

Structure Cell wall: Protection from osmotic stress, helps maintain cell - One genuine cell wall made of -Plasma membrane
shape. cellulose, or hemicellulose -Cell wall: made of chitin
2 main types: surrounds the yeast's body's one -Cytoplasm : Mitochondria,
 Gram(+): 1 layer (20-80nm), made of peptidoglycan. cell. ER, Golgi apparatus,,
Components: - cell wall: beta-glucan and inclusion, lysosome,
Peptidoglycan: 30-95% galactomannan peroxisome
Teichoic acid: High - Mitochondria Organelles:
Lipopolysaccharides: 0% - Cytoplasm - Endoplasmic reticulum
Protein: 0%  Cytoskeleton: 3 types of - Golgi apparatus
 • Thick peptidoglycan filaments form: - Nucleus, nuclei
• Teichoic acids microfilaments, -Vacuole
• Disrupted by lysozyme microtubules, intermediate - Ribosome: 2 types: 80S
• High Penicillin sensitive filaments and 70S (same as bacteria)
• Produce exotoxins - Ribosomes: 70S, 80S = dimer of Cytosol:
 Gram(-): 2 layers (2-7nm, 7-8nm), made of a 60S and a 40S subunit, has a -Cytoskeleton:
peptidoglycan, periplasmic space. sedimentation coefficient of 80S microfilaments and
Components: - Nucleus intermediate filaments
Peptidoglycan: 5-20% - Golgi apparatus
Teichoic acid: 0% - Plasma membrane( the same
Lipopolysaccharides: 20% mozaic model and function with
Protein: High bacteria and mold) structure diff is
• Thin peptidoglycan symmetric and asymmetric
• Outer membrane - Vacuole
• Periplasmic space - Bud Scar
• Tetracycline sensitive - Peroxisome
• Produce Endotoxins and Exotoxins - Periplasm
- Endoplasmic reticulum
Plasma membrane: - Centrioles
- Function: Selectively permeable barrier, mechanical boundary
of cell, nutrient and waste transport, location of many metabolic
Structure processes (respiration, photosynthesis), detection of
environmental cues for chemotaxis

- Structure: phospholipid is symmetric

- Components:
• Bilayer phospholipid
membrane that encompasses
the cytoplasm
• Thin layer about 5-10 nm
• Fluid mosaic model
• Composed of lipid, protein
(peripheral proteins and
integral proteins),
oligosaccharides
• No sterol, but hopanoid
Cytoplasm and other organelles
1. Nuclear body (thể nhân)
2. Cytoplasm (tế bào chất):
Function:
• Matrix lying between the plasma membrane and the nucleoid
 • Is largely water (about 70% )
• Lacks unit membrane-bound organelles
• Often is packed with ribosomes and highly organized
• Medium for internal cell organelles
(protein, DNA, ribosomes, inclusion bodies ...)
Structures:
+Organelle structures:
Nucleoid:
• Is an irregularly shaped
region where located
bacterial chromosome
• Attached to plasma
membrane
• Contains single circle of
double-stranded
deoxyribonucleic acid (DNA)
(some have a linear DNA chromosome.
Vibrio cholerae have more than one
chromosome)
• Composed of about 60%
DNA, 30% RNA, and 10%
protein by weight
Plasmids:
• Locate in cytoplasma
• Do not attached to plasma
membrane (Float freely in
cytoplasma)
• Are circular double-stranded
deoxyribonucleic acid
(DNA)
• Able to replicate separately
with chromosome
• Able to be lost during cell
division
• Not required for host
Structure growth and reproduction
Inclusion (hạt dự trữ):
• Located in cytoplasma
• Storage granules of organic and inorganic materials
• Normally bounded by a single layer membrane (2-4 nm) made
from protein or phospholipid
Ribosome:
• Located in cytoplasma and may linked to plasma membrane:
matrix ribosomes and plasma ribosomes
• Called 70S ribosome, size about 14-15 x 20 nm
Svedberg unit - unit of the sedimentation coefficient
• very complex objects made of both protein and ribonucleic
acid (RNA)
• Constructed of 02 sub-units: 30S sub-unit + 50S sub-unit
• site of protein synthesis
matrix ribosomes synthesize proteins destined to remain within
the cell
plasma membrane ribosomes make proteins for transport to the
outside.
+Cytosol: Bacterial cytosol is not as simple as once thought
Water
Cytoskeletal proteins
3. Vacuole (không bào): An inclusion that provides buoyancy
for floating in aquatic environments.
4. Spore, endospore (bào tử): Survival under harsh
environmental conditions

External structure:
10. Capsule/ Slime layer (màng nhầy/ lớp nhầy): Resistance to
phagocytosis, adherence to surfaces
11. S-Layers
12. Flagella and Fimbriae /pilus (tiên mao, khuẩn mao):
Swimming and swarming motility; Attachment to surfaces,
bacterial conjugation and transformation, twitching
 Chap 3: Cell requirements

Chemical requirement (Nutrition requirements): Physical requirements

 Oxygen Aw
Structure organic compound (carb, lipid, nucleic acid and pH
protein) and inorganic salts Temperature
Enzymatic functions O2
 Hydrogen
Pressure
From all organic compounds and several inorganic
Radiation
compounds such as water
Role: pH, hydrogen bond, respiration
 Carbon
Role: skeleton of all organic molecules
 Phosphate
Phospholipid in cell membranes; and coenzymes such as
NAD+
Energy: ATP
 Sulfur
Vitamins, amino acids (methionine, cysteine)
disulfide bonds (protein 3 dimensional structure)
 Nitrogen
indispensable structure to the structure of protein, DNA,
RNA, ATP)

Bacteria 1. Divided as quantity of nutrient - Radiation:

Photooxidation: visible light
Macronutrient: protein, carb Oxygen quenching colorants
 required in large quantities, play principal roles in cell Ionizing radiation: causes a variety of changes in cells:
structure and metabolism • breaks hydrogen bonds
• oxidizes double bonds
Micronutrient: Mn, Zn, Ni • destroys ring structures
 required in small amounts, involved in enzyme • polymerizes some
function and maintenance of protein structure molecules
Ultraviolet (UV) radiation:
Trace elements: K, Na, Ca, Mg, Fe • short-UV (260 nm): most effectively absorbed by and damaging to
 Inorganic elements required in small amounts DNA
• near-UV (325 to 400 nm): induce the breakdown of tryptophan to toxic
2. Divided as types of nutrient photoproducts produce breaks in DNA strands
Visible light: give energy to photosensitizers (chlorophyll,
bacteriochlorophyll, cytochromes, and flavins) = photooxidation
Inorganic nutrient: metal and their salt, (magnesium
MOs use of carotenoids to quench singlet oxygen
sulfate, ferric nitrate, sodium phosphate, sulfur hydro - H S),
2
gases (oxygen, carbon dioxide) and water
Organic nutrient: CH4, carbohydrates, lipids, proteins and - Water activities- osmosis effect on microbial cells.
nucleic acids. + Halophile
Nutritional type of microbe: Requires high levels of sodium chloride, usually above about 0.2 M to
Carbon sources: grow
 Heterotroph: mus obtain carbon in an organic form: + Osmotolerant
protein, lipids, nucleic acid Able to grow over wide ranges of water activity or osmotic
 Autotroph : use CO2 and not dependent on other concentration
living things + Xerophile
Energy sources: Organisms that grow best at low water activity, typically with optima at
 Phototrophs: derive energy from sunlight 0.85 or below
 Chemotrophs: obtain energy from chemical reactions Require more water than Yeast
Electron sources:
 Lithotrophs: use reduced inorganic substances as - pH: Maintain the intracellular pH; Prevent protein denaturation
electron sources + Bacteria: pH 6.5 - 7.5
 Organotrophs: extract electrons from reduced Acidophile
organic compounds Growth optimum between pH O and 5.5
15 and 8.0
Chemoorganotrophic heterotrophy: Fungi Neutrophile
Chemolithotrophic autotrophy: Bacteria Growth optimum between pH 5.5 and 8.0
Photoorganotrophic heterotrophy: Bacteria Alkaliphile
Photoorganotrophic autotrophy: Bacteria Growth optimum between pH 8.0 and 11.5

Growth factor: amino acid, nitrogenous base, vitamin. Only - Temperature ( effect both function and cell structure )
heterotrophs require . Cardinal temperature of a microbe
Classification : • Minimum temperature – lowest temperature that permits a microbe’s
 Essential amino acids : protein synthesis growth and metabolism
 Purines and pyrimidines : nucleic acid synthesis • Maximum temperature – highest temperature that permits a microbe’s
 Vitamins : enzymes cofactors synthesis growth and metabolism
• Optimum temperature – promotes the fastest rate of growth and
metabolism
• Psychrophile: optimum growth temperature of 15C
• Psychrotroph
• Mesophile: growth optima around 20°C to 45°C
• Thermophile: grow at temperatures
between 45°C and 85°C, and they often have optima between 55°C
and 65°C.
• Extreme thermophile

• MOs cannot regulate their internal temperature

• Sensitivity of enzyme-catalyzed reactions
• At low temperatures: optimum temp
A temperature rise increases the growth rate because the velocity of an
enzyme-catalyzed reaction
• At high temperatures:
Denaturing enzymes, transport carriers and other proteins.
Disruption of membranes
Melting and disintegration of the lipid bilayer
• At very low temperatures:
Solidification of membranes solidify and cessasion of enzymes activity

- Oxygen:
Obligate aerobe
Completely dependent on atmospheric O2, for growth
Facultative anaerobe
Does not require O2, for growth but grows better in its presence
Aerotolerant anaerobe
Grows equally well in presence or absence of O2
Obligate anaerobe
Does not tolerate O2, and dies in its presence
Microaerophile
Requires O2, levels between 2-10% for growth and is damaged by
atmospheric O2 levels (20%)

- Pressure:
• Barophile (piezophile):
Growth more rapid at high hydrostatic pressures

Osmotic Pressure
• Hypertonic environments, or an increase in
salt or sugar, cause plasmolysis
• Hypotonic: the solute concentration of the external environment is
lower than that of the cell’s internal environment.
• Isotonic: the environment is equal in solute concentration to the cell’s
internal environment

• Extreme or obligate halophiles require high

osmotic pressure
• Facultative halophiles tolerate high osmotic
pressure

MOs living in deep sea undergo a hydrostatic pressure that can reach
 600 to 1,100 atm and the temperature is about 2-3 C. These high
o

hydrostatic pressures affect membrane fluidity and membrane

associated function.

What is the difference between Barotolerant and Piezophilic

(barophilic)?
increase the amount of unsaturated fatty acids in their membrane lipids
as pressure increases.
may also shorten the length of their fatty acids.

Yeast Chemical requirement Physical requirements

Chemical energy source (organic) Oxygen :
Organic H donor  When yeast respires, O2 will be consumed. CO2 is produced
Organic C source  Fermentation : Yeast need anaerobic environment
The energy source is alcoholic fermentation, which Temperature :
produces ethanol and carbon dioxide as the end product  The optimum temperature for growth of yeasts is in the
of carbohydrate metabolism. mesophilic range of 25–30 °C.
 Yeasts in general can grow over a range of temperatures from 0
°C to 47 °C.

pH (minimum, maximum, optimum):

 Yeasts grow well under acid conditions, at pH 4.0–4.5.
 They can grow at lower pH than most bacteria, but do not grow
well under alkaline conditions
Water : Need plentiful water supply, same osmotic pressure as
bacteria
Pressure : Ideal pressure is 10PSI - 12PSI
(1 psi = nearly 6900 Pa)

Mold Chemical requirement Physical requirements

Chemical energy source (organic) pH : Molds grow between pH 5 and 6
Organic H donor Temperature: Most molds cannot grow below 4.4 °C. Mold grows best
Organic C source between 77° F and 86° F (25°C - 30°C), especially if the air is humid.
The molds produce a hydrolytic enzyme that degrades Water: Mold will grow in places with a lot of moisture but less water
the biopolymers like starch and cellulose into simpler supply than Yeast, same osmotic pressure as bacteria
carbohydrates. Oxygen: obligate aerobes.
These simpler compounds are then absorbed by the cell. This means that they need oxygen to survive. Mold grows even at very
Vitamin low concentrations of oxygen, however, which makes it difficult to fight
mold growth by limiting oxygen
 Chap 3: Microbial growth and control
Pattern of microbial growth and death
growth control

Mode of reproduction: (asexual) death pattern

Pattern  binary fission: bacteria • Death is a phenomenon that involves the permanent termination of an
 budding: yeast organism’s vital
 Reproductive spores: mold processes. Death means the irreversible loss of the ability to reproduce (grow
(conidiospores, sporangiospores, and divide)
and arthrospores) • The permanent loss of reproductive capability, even under optimum growth
Growth curve (model): conditions,
1. Lag phase – “flat” period of adjustment, has become the accepted microbiological definition of death.
enlargement; little growth − Permeability of plasma membrane
2. Exponential growth phase – a period of − Enzyme activities
maximum growth will continue as long as − Cultivability
cells have adequate nutrients & a favorable • A microbial population is not killed instantly when exposed to a lethal agent.
environment (µ = µmax) • Population death is generally exponential (logarithmic);
3. Stationary phase – rate of cell growth the population is reduced by the same fraction at constant intervals
equals rate of cell death cause by depleted Microbial death pattern
nutrients & O excretion of organic acids & Bacterial populations usually die at a constant rate when heated or when
pollutants (µ = 0) treated with antimicrobial chemicals.
4. Death phase – as limiting factors Figure 2
intensify,
cells die exponentially in their own wastes Parameters to measure an antimicrobial agent’s killing efficiency
5. Long-Term Stationary Phase - some Decimal reduction time (D) or D value:
microbes have a long period where the − is the time required to kill 90% of the microorganisms or spores in a sample at
population size remains more or less a specified temperature
constant. − used to estimate the relative resistance of a microorganism
Figure 1 Figure 3

Factors influence antimicrobial agent’s killing efficiency

1. Population size (Number of microbes)
More microbes mean longer time to die because die with a constant rate
2. Population composition (Nature of microbes in
the population)
3. Temperature & pH of environment
Higher temperature will reduce the time
4. Concentration of agent (dosage, intensity, time
of exposure)
For example, UV radiation is most effective at 260 nm, and most disinfectants
are more active at higher concentrations.
 5. Mode of action of the agent
6. Presence of solvents, organic matter, or inhibitors
Inhibitors will slow down the time of killing. For example, saliva, blood, and
feces can inhibit the actions of disinfectants and even of heat.
7. The type of microbial growth

Techniques to Direct Methods

count the -Plate counts
number of - Take 1 ml of sample solution, dilute to the required concentration, take the inoculum on a petri dish, wait for the colonies to
microbes grow.
(enumeration/ count colonies
measure the - Advantages:
cell density) + is the standard method , used to verify other methods
- Defect:
+ Depends on the technique of the performer
The diluent must be homogeneous between each dilution
+ Takes a long time to wait for colony growth
+ Colony count results are not current values
+ Only detects cells that are still alive

-Filtration
Filter the suspension containing microorganisms through multiple filter funnels, the funnels and racks must be sterile
- Advantages: quickly check the density of microorganisms in the suspension with low microorganisms concentration
- Defect:
+ an only count suspensions with few microorganisms
+ The suspension solution containing microorganisms to be tested must have a small viscosity
+ Only Identify Living Cells
+ Not suitable for analysis of solid food samples

-MPN
- Buy a set of tubes, put the sample into the tube and leave for a period of time depending on the type of microorganism. Then
observe the color and check the results according to the sample table attached to the set of tubes
- Advantages: quantification of microorganisms density in suspensions with low microbial density
- Defect:
+ can only be used once
+ the result is not very accurate
+ takes a lot of time to wait for microorganisms to develop
+ only counts live cells
+ the need for large numbers of replicates at the appropriate dilution to narrow the confidence intervals.

In this method, numerous replicates of several dilutions of a culture are made and added to tubes containing a suitable liquid
growth medium. After incubation, each tube is examined to determine if growth occurred. It is assumed that the last tube in the
dilution series that demonstrates growth was inoculated with between one and 10 cells, while the next tube had between 11
and 100 cells, and so on. If no growth is observed, then the tube is assumed not to have received any cells. In this way, the
number of cells in the original sample is estimated.

-Direct microscopic count: A small sample is placed on the grid under a cover glass. Individual cells, both living and dead,
are counted. This number can be used to calculate the total count of a sample. Counting can be automated by sensitive
devices such as the Coulter counter, which electronically scans a culture as it passes through a tiny pipette.

-Flow cytometry: Fluorescently labeled cell samples are injected into a fluid stream that narrows using a technique called
hydrodynamic focusing. This forces only one cell at a time to pass a laser beam and scatter light in all directions. Forward-
scattering light is a function of cell size, while the nature of laterally scattered light is determined by morphological complexity.
This information is detected, measured, and converted to an electrical signal that reports the number and types of cells
present.

Indirect Methods
- Turbidity: measure light absorption. They depend on the fact that microbial cells scatter light that strikes them. The amount
of scattering is directly proportional to the biomass of cells present and indirectly related to cell number.
Advantages: rapid and sensitive.
Defect:
Can't distinguish between living or dead cells.
Cannot apply for milk detection as it is not transparent enough to measure the light absorption.
phai tu lap duong chuan
The population needs to be high enough to give detectable turbidity.

- Metabolic activity : dùng máy đo mật độ ATP

Advantages: Saving time, quick method, easy to use
Disadvantages: Expensive ,can't distinguish between living or dead cells.

- Dry weight : used for filamentous organisms such as fungi

Advantages:
+ immediately determine the density of microorganisms, no waiting time
+ easy to do
Defect:
Can't distinguish between living or dead cells.
 Chap 3: Methods to remove or control a microbial population
method to grow/control is not about the requirements.
Media: Level to control:
-Yeasts can flourish in a variety of settings, including the skin and guts of Sterilization: disinfection and antiseptics
mammals, fruits, and berries. - Complete removal of all viable microorganism (include virus and
- Molds require damp and moist places for growth. spores). Not always practical. because may be not destroy no-spore
Temperature: Disinfection: removal of vegetative pathogens but no bacteria
-The ideal temperature for mold’s growth is 37℃. endospores. on inanimate objects
- The ideal temperature for mold’s growth is 28℃. antiseptics: destroy or inhibit vegetative pathogens on living tissue
pH : Decontamination: sanitization and degermation
-Molds grow between pH 5 and 6 sainitization: reduce to a public health standard
-Yeasts grow well under acid conditions, at pH 4.0–4.5. degermination: remove microbe form a limited area
 They can grow at lower pH than most bacteria, but do not grow Use biocide or bacteriostasic (to kill or inhibit, respectively)
well under alkaline conditions Methods to control
Oxygen : Physical methods:
-When yeast respires, O2 will be consumed. CO2 is produced  Heat
-Fermentation : Yeast need anaerobic environment Moist heat and dry heat
-obligate aerobes. In moist heat: use hot water or steam
This means that they need oxygen to survive. Mold grows even at very +Pasteurisation: kill vegetative but not spore
low concentrations of oxygen, however, which makes it difficult to fight +Boiling 100C for 30 mins to destroy non-spore forming
mold growth by limiting oxygen pathogens
Solute Concentration and Water Activity: +Tydallisation: successive pasteurization, destroy spores
+ Halophile: Requires high levels of sodium chloride, usually above about Dry heat use more temperature than moist heat
0.2 M to grow  Filtration: Physical removal of microbes by passing a gas or liquid
+ Osmotolerant: Able to grow over wide ranges of water activity or through filter
osmotic concentration  Low temperatures: Slows the growth of microbes
+ Xerophile: Organisms that grow best at low water activity, typically with  High pressure: Alters the molecular structures of proteins and
optima at 0.85 or below carbohydrates
Pressure  Desiccation: Gradual remove of water from cells, leads to
+Piezophile: Growth more rapid at high hydrostatic pressures metabolic inhibition
Method  Osmotic pressure: Decrease the moisture needs for cells growth
+Closed system (batch culture): no food added, no wastes remove)  Radiation
 Lag phase: adaptation period when microbes are adjusting in + Ionizing radiation: Radiation of very short wavelength and
their new condition high energy → Causes a variety of changes in cells ( breaks
 Log phase (exponential phase): is marked by predictable hydrogen bonds, destroys ring structures…)
doublings of the population + Non ionizing radiation: (Ultraviolet radiation) → Damage to
 Stationary phase: At this point the number of new cells being DNA in the form of thymine dimer mutations
produced is equal to the number of cells dying off Chemical method: chap 3- Methods to remove or control a microbial
 Death phase: the number of viable cells decreases in a population – page 25-34
predictable (or exponential) fashion. Biological methods: control of MOs by natural such as predation,
+Continuous culture: food added, wastes remove, viral lysis, and toxins
 Purpose: Control in density of the cells (population)