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Bacteria

Smallest free-living organism


A prokaryote that lacks true nucleus
Represent a large diverse group of organism

Bacterial diversity and ubiquity


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Bacteria are diverse in shape and size, adaptation to environment, and metabolic
processes
Descriptive term

Adaptive feature

Psychrophile

Growth range -40C to +20C

Mesophile

Growth range +20C - +40C

Thermophile

Growth range +40C - +85C

Thermoduric

Endure high temperatures

Halophile

Salt-tolerant

Acidophile

Acid-tolerant

Aerobe

Air (oxygen) requiring

Obligate anaerobe

Air (oxygen) poisoned

Autotroph

Utilizes inorganic material

Heterotroph

Requires organic material

Cell size and shape


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Majority of bacteria are 1-5 nm long and 1-2nm in diameter


Classification of bacteria is made through morphological grounds
Comma shape (Vibrio), Loosely coiled (Spirillum) , Tightly coiled (Spirochete)

Cell wall
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Maintains the shape and integrity of the bacterial cell


A target for antibiotics (cell lysis)
Provide strong, rigid structural component to withstand osmotic pressures
Most bacterial cell walls have PEPTIDOGLYCAN layer (murein)
Peptidoglycan is composed of N-acetyl muramic acid and N-acetyl
gluosamine

Gram stain

Differentiates gram-negative bacteria and gram-positive bacteria

Steps:
1.
2.
3.
4.

Application of crystal violet


Application of iodine
Alcohol wash
Application of safranin

Ex: gram-positive (staphylococcus aureus), gram-negative (Escherichia Coli)

Gram-positive wall
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Contains techoic acids and lipotechoic acids


During an infection, lipotechoic acid molecules trigger an inflammatory
response.

Gram-negative cell wall


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Compose of proteins, lipoproteins, phospholipids, and lipopolysaccharide


(LPS) that is unique to gram negative bacteria
LPS determine the antigenicity of bacteria
LPS is made up of lipid A, core polysaccharide and O-specific
polysaccharide
Lipid A is responsible for the toxic properties of gram negative bacteria

Cytoplasmic membrane
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Involved in various transport and enzyme functions associated with the


membrane
Transport of nutrients, energy regeneration, and electron transports
Selective barrier between the cytoplasm and the cell environment.

Cytoplasm

- Consists of 80% water and contains enzymes that generate ATP


- Compose of the ribosomes, nucleoid and inclusion granules
Nucleoid

- Singular, closed circular molecule of double-stranded DNA


Ribosomes

Site of protein synthesis


Two subunits: 30s and 50s

Inclusion granules

- Serves as the storage material for carbon, nitrogen and Sulphur


Flagella
a. Monotrichous a single polar flagellum
b. Lophotrichous two or more flagella at one pole of the cell.
c. Amphitrichous single/tuft of flagella at each end of the cell.
d. Peritrichous flagella distributed over the entire cell.
Pili and Fimbriae
Function:
a. Fimbriae for adherence of cells to one another and to environmental surfaces.
b. Pili (pilus) join bacterial cell in preparation of DNA transfer from one cell to
another.
Glycocalyx (Slime layer and capsule)

- General substances that surround cells


- Gelatinous polymer of polysaccharide, polypeptide, or both.
- Functions as protective barrier
Biofilms

- Any surface for microbial habitat


- Usually contains more than one species of bacteria which exist and cooperate
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together
Biofilm formation begins with attaching to surface and form a cement cells to the
surface to protect the bacteria from hazardous materials

Bacterial sporulation

- Not part of the reproductive cycle but enables the bacteria to survive in
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adverse environmental conditions


All sterilization process for pharmaceutical products have been designed to
destroy bacterial spore

Bacterial toxin

- Normal flora bacteria that are not pathogenic but can be opportunistic
- Pathogenic bacteria that is not a normal flora and cause diseases
- Toxins products of bacteria that produce immediate host cell damage
Endotoxin

The lipid A component of LPS


Ability to induce fever and activate B lymphocytes
Generally released from lysed or damaged cell

Exotoxin

A-B toxin contains B subunit that binds to host cell receptor and bound to the A
subunit that mediates enzyme activity for toxicity

Cytolytic toxins does not have A and B subunit but work enzymatically to
attack cell that causes cell lysis

Superantigen toxin stimulate large number of immune response cell resulting to


massive inflammatory response

Multiplication and Division cycle

Bacteria multiply in number by binary fission


Each daughter cell contains same material of the mother cell
DNA replication is dependent on temperature
Example: E. coli grows at 37C and replication of chromosomes will take
approximately 45 minutes
The time interval between one cell division and the next is called the Generation
time

Traditional cultivation
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Cultivation (in vitro) - A process of growing microorganisms in culture. Meet its


nutritional and environmental requirements

Phases of culture media:


1. Broth (liquid) nutrients dissolve in water
2. Agar (solid) with solidifying agent

3. Biphasic with solid and liquid phase


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General concepts of Culture media

Types of bacteria by nutrient requirements

1. Fastidious
- Complex or extensive nutritional requirements
- Culture Media should be supplemented
2. Non fastidious
- Basic nutritional requirements

- Media Classifications and Functions

1. Supportive media- Support growth of most non fastidious organisms. (Nutrient


agar contains beef extract , peptone, water, agar)
2. Enriched media (Non-selective isolation media) - nutrients supplement added to
supportive media for growth of fastidious microbes
3. Selective media - Permits growth of certain bacteria while inhibitory to others.
4. Differential media - Provides a distinct cultural appearance of microorganisms.

Microbial growth and nutrition

Bacterial growth
Growth curve
a. Lag phase little or no cell division; intense metabolic activity
b. Log phase - AKA Exponential growth phase; cell begins to divide; active
cellular reproduction with constant minimum generation time; cells are at their
most active state
c. Stationary phase - -growth rate slows down (# of new cells = # of microbial
deaths = population stabilizes) period of equilibrium
d. Death phase - logarithmic decline; number of deaths exceeds the number of
new cells formed