CHAPTER 3: BACTERIA

Molina, Gernelyn Shaira C.

BACTERIA
- smallest free-living organisms
- prokaryote (lacks true nucleus)
BACTERIAL DIVERSITY AND UBIQUITY
-can be seen in terms of variation in cell size and shape (morphology), adaptation to
environmental extremes, survival strategies and metabolic capabilities.
Hot Sulphur springs (65C) - deep freezers (-20C)
High (pH1) - low (ph13) acidity
High (0.7m) low osmolarity (water)
Nutritionally rich (compost) Nutritionally poor (distilled water)
Psychrophile
Mesophile
Thermophile
Thermoduric
Halophile
Acidophile
Aerobe
Obligate anaerobe
Autotroph
Heterothroph

-40C to 20C
20C to 40C
40C to 85C
Endure high temp.
Salt-tolerant
Acid-tolerant
Air(oxygen) requiring
Air(oxygen) poisoned
Utilizes inorganic
material
Requires organic
material

CELL SIZE AND SHAPE

-1-5 m long, 1-2 m in diameter
Thiomargarita namibiensis - >5 m in diameter
-unicellular
-round (cocci), cylindrical (rod) or bacillus, ovoid
Vibrio - curved
Spirochaetes -longer rigid curved w/ multiple spirals
Actinomycetes rigid bacteria resembling fungi
CELL WALL
Primary function provide a strong, rigid structural component that can withstand
the osmotic pressures caused by high chemical concentrations of inorganic ions in
the cell.
-Peptidoglycan (murein or glycopeptide) -large molecule containing glycan
(polysaccharide) chains that are cross-linked by short peptide bridges.
- N-acetyl muramic acid and M-acetyl glucosamine
Glycan chains- backbone to peptidoglycan
GRAM STAIN
-differential staining technique
Gram positive Staphylococcus aureus

Gram negative Escherichia coli

GRAM POSITIVE CELL WALL

-60-80% peptidoglycan
-teichoic acids and lipoteichoic acids
-during an infection, lipoteichoicacids trigger an inflammatory response
Teichoic acids- either ribitol phosphate or glycerol phosphate molecules connected
by phosphodiester bridges
-responsible for the negative charge
GRAM NEGATIVE CELL WALL
-10-20 % peptidoglycan
-proteins, lipoproteins, phospholipids and lipopolysaccharide (LPS)
LPS determines the antigenicity of the bacteria
- made up of lipid A, core polysaccharide and O-specific polysaccharide
Lipid A responsible for toxic and pyrogenic properties of the gram negative
bacteria
-Linked to the core polysaccharide by ketodeoxyoctonate (KDO)
Abequose unusual deoxy sugars at O- polysaccharide
CYTOPLASMIC MEMBRANE
-transport nutrients, energy generation and electron transports
- involves in various transport and enzyme functions associated w/ the membrane
- Selective barrier between the cytoplasm and the cell environment
Mesosomes invaginations of the cytoplasmic membrane
CYTOPLASM
-80% water
-contains enzymes that generate ATP
Nucleoid singular, closed circular molecule of double-stranded DNA
Plasmids circular pieces of double-stranded extrachromosomal DNA
-capable of autonomous replication
Ribosomes site of protein synthesis: two subunits: 30S and 50S

Inclusion granules storage material for carbon, nitrogen and Sulphur

FLAGELLA
-bacterial motility
-project from the surface of the cell
FIMBRIAE adherence
-responsible for haemagglutination and cell clumping
-type I fimbriae of enteric(intestina) bacteria- best characterized
PILI join (cell-cell)
-involved in the genetic exchange process of conjugation
GLYCOCALYX (slime layer and capsule)
-general substances that surrounds cells
-gelatinous polymer of polysaccharide, polypeptide or both
a. Capsule organized, firmly attached to the cell wall
b. Slime layer unorganized, loosely attached to the cell wall
SAME FUNCTION:
-to offer protection against desiccation
-to provide a protective barrier against the penetration of biocides, disinfectants and
positively charged antibiotics
-to protect against engulfment by phagocytes and protozoa
-to act as a cement binding cells to each other and to the substratum in biofilms
Polymers that performs all these functions:
Alginate Pseudomonas aeruginosa
Dextran Leuconostoc mesenteroides
EXTRACELLULAR POLYSACCHARIDES (EPS)
-2% carbohydrate 98% water
-provides a gummy exterior to the cell
S-LAYERS - most common cell wall type amongst the archaea
-external permeability barrier
BIOFILM any surface for microbial habitat
BACTERIAL SPORULATION not part of reproductive cycle but enables the
bacteria to survive in adverse environmental conditions
-All sterilization process for pharmaceutical products have been designed to destroy
bacterial spores
ENDOSPORE STRUCTURE
ENDOSPORE differentiated cells that possess a grossly different structure to that
of the parent vegetative cell in which they are formed
-presence of dipicolinic acid and high levels of calcium ions
-the core is partially dehydrated (10-30% of water content of vegetative cells)
Dehydration- increase resistance to both heat and chemicals
BACTERIAL TOXINS
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
-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 enzymes activity for toxicity
Cytolitic toxins does not contain A-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
Ex. E. coli 37C 45 mins
Generation time- time interval between one cell division and the next
1.
2.
3.
4.

Cell elongates and DNA is replicated

Cell wall and plasma membrane begin to divide
Cross-wall forms completely around divided DNA
Cells separate

TRADITIONAL CULTIVATION
Cultivation process of growing microorganisms in culture
-meet its nutritional and environmental requirements
PHASES OF CULTURE MEDIA
1. BROTH (liquid) nutrients dissolved in water
2. AGAR (solid) with solidifying agent
3. BIPHASIC with solid and liquid phase
Broth- 3 mL, Slant- 6 mL, Deep tube- 10 mL
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 CLASSIFICATION AND FUNCTIONS
1. Supportive Media support growth of most non-fastidious organisms
- beef extract, peptone, water, agar
- nutrient agar
2. Enriched Media (non-selective isolation media)
-nutrients supplement added to supportive media for growth of fastidious organisms
- SBA, CAP (chocolate agar plate)
3. Selective Media permits growth of certain bacteria while inhibiting to others
- HE AP, SS AP

4. Differential Media promotes a distinct cultural appearance of microorganism

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

TRANSFORMATION
- when a naked DNA molecule from the environment is taken up by the cell and
incorporated into its chromosome in some heritable form
TRANSDUCTION
- the transfer of bacterial genes by viruses (bacteriophages); it occurs as the result
of the reproductive cycle of the virus
CONJUGATION
- the transfer of genetic information via direct cell-cell contact; this process is
mediated by fertility factors (F plasmids)
PHYSICOCHEMICAL FACTORS THAT AFFECT GROWTH AND SURVIVAL OF
BACTERIA
1. Temperature
2. pH
3. Water activity/ solutes
4. Availability of oxygen