BACTERIAL ANATOMY &

PHYSIOLOGY
DENNIS R. ESQUIVEL,
RMT, MD, MPH, DPASMP,CSPSH, FPAFP
Department of Microbiology & Parasitology
Remedios Trinidad Romualdez Medical Foundation
College of Medicine
DISTINGUISHING CHARACTERISTICS
Prokaryotes Eukaryotes
Size: 1 – 10 um 10 – 100 um

No nuclear membrane Nuclear membrane present

Single chromosome Multiple chromosome

No DNA-associated histones Histones associated with DNA

Binary fission Mitotic division

Lack membranous compartments Have membranous compartments

Peptidoglycan (cell wall) Chitin or Cellulose in the cell wall

Steroids absent Steroids maybe present

70s Ribosomes 80s Ribosomes

Anaerobic respiration possible No anaerobic respiration except some

yeasts
BACTERIAL STRUCTURE
2 Parts:
1. Cell envelope
2. Cytoplasmic components
BACTERIAL STRUCTURE

Cell Envelope:
 Cell wall

 Cytoplasmic Membrane
CELL WALL

 Peptidoglycan: principal structural component

 Thicker in Gram (+) organisms (40 sheets)
 Hydrolyze by Lysozyme through cleavage of the glycosyl bonds
 Autolysin: dissolve the peptidoglycan layer necessary for:
 Cell wall growth
 Cell septation
 Sporulation
 Competency for transformation
CELL WALL
Functions
1. Maintains the morphology of the cel
2. Allows the cell to withstand osmotic pressure up
to 20 atmospheres
3. Site for various antigenic determinants
4. Source of endotoxin for Gram (-) organism
CYTOPLASMIC MEMBRANE
“Plasma membrane”
 The physical & metabolic barrier of the cell

 Functions:

1. selective permeability & transport of the solutes into

the cell
2. Electron transport & oxidative phosphorylation
3. Secretion of hydrolytic enzymes
4. Biosynthetic functions:
a. Site for assembly of carrier lipids for cell wall
synthesis & site for enzymes involved in cell wall
synthesis
b. Site for enzymes involved in phospholipid synthesis
MESOSOMES
 Complex invaginations of cell membrane
 Has role in reproduction of the cell

 First site for cell division to occur

DIFFERENCE BETWEEN GRAM (+) & (-)
CELL WALL
 Gram (+) organism cell wall:
 Have simplier but thicker cell wall made up of
multiple layers of peptidoglycan layer with
teichoic acid polymers
DIFFERENCE BETWEEN GRAM (+) & (-)
CELL WALL
Gram (-) organism cell wall:
 Thinner wall ( 5 sheets only of peptidoglycan)

 No associated Teichoic acids

 Has an outer membrane on top of the

peptidoglycan layer
 Outer membrane: thicker than the peptidoglycan
 Composed of:
 Lipid bilayer
 Proteins

 Lipopolysaccharides (LPS)

 Endotoxin
DIFFERENCE BETWEEN GRAM (+) & (-)
CELL WALL
Gram (-) organism cell wall:
 Lipid Bilayer: attached to the Peptidoglycan
layer by lipoproteins that cross the periplasmic
space
 Proteins: “Porins”
 Form as transmembrane channels
 Transport of Ions & hydrophilic compounds from
extracellular compartment to the periplasm
DIFFERENCE BETWEEN GRAM (+) & (-)
CELL WALL
Gram (-) organism cell wall:
 Lipopolysaccharides:

 Composition:
 Lipid portion (Lipid A)
 Polysaccharide-rich core
 Polysaccharide side chain

 Polysaccharide portion: antigenic & designated

as O antigen
 Lipid portion: Heat stable – responsible for the
biologic effects of Endotoxin
BACTERIAL LYSIS
 Results from the removal of the cell wall
 Effects on Gram (+) cell wall:
 Protoplast: results from complete removal of cell wall
 Composed of cytoplasmic membrane & bacterial
contents
 Require ISOTONIC SOLUTION to assume a
spherical configuration
 Can not maintain integrity in a hypo- or hypertonic
medium
BACTERIAL LYSIS
 Results from the removal of the cell wall
 Effects on Gram (-) cell wall:
 Gram (-) organism: resistant to enzymatic
destruction because of the complex cell wall
 Spheroplasts: cells with damage cell wall
 Still assume spherical shape even in non-isotonic
medium
CYTOPLASMIC COMPONENTS
 Composed of Nuclear body or Nucleoid,
Ribosomes, Endospores & Granules
Nucleoid:
 DNA concentrated in the cytoplasm
 Consists of one double – stranded, circular,
covalently closed, supercoiled DNA molecule

Plasmids
 Small amount of DNA that persists as
extrachromosomal element
 Code for genes that determine virulent behavior
of the microorganisms
CYTOPLASMIC COMPONENTS
Ribosomes:
 Complex globular structures composed of RNA
molecules & associated proteins
 70 Svedberg units (70s) (sedimentation coefficient)

 Composed of 2 sub-units: 50s & 30s

Storage Granules:
 Hold temporarily excess metabolites

 Their presence & amount vary with species of

bacteria & its metabolic activity
 Ex: Babes Ernst Granules or Volutin- present in
Corynebacterium diphtheriae
CYTOPLASMIC COMPONENTS
Endospores
 Resistant to heat,
drying & chemical
agents

External Structures:
1. Capsule
2. Flagella
3. Pili
CYTOPLASMIC COMPONENTS
Capsule:
 Composed of complex polysaccharides

 Visualized through Quellung (Swelling) Reaction

Antibody specific
Culture
For the Capsule in ?

Binding of antibody to the capsule; there is change in light defracting properties

So that the capsule will appear as swollen
CAPSULE
 Polysaccharide in nature except for Bacillus anthracis which
have polypeptide capsule composed of D – Glutamic Acid
 D-amino acid isomer: Resistant to proteolytic enzymes
thereby preventing phagocytosis
 It can be visualized through NEGATIVE staining technique
 Capsule appear as “halo” surrounding the cell
 Stains: Nigrosin, India ink technique
CAPSULE
Functions:
1. Protects the cell from phagocytosis
2. Prevents some viruses to attach to the cell wall
3. Prevents penetration of antibiotics

Flagella:
 Organ for locomotion

 “whip – like” structure

 Composed of Flagellin
PILUS
 Protein fibers that cover entire surface of Gram (-)
organism
 Nomenclature between types depends its functions
 Evasin: prevent phagocytosis
 Adhesin: prevent adhesion
 Aggressin: Leukocidal
 Sex Pilus: Reproduction; causes drug resistance

 Function: play a role in bacterial adherence to mucosal

surfaces which is essential step for colonization
BACTERIAL PHYSIOLOGY
NUTRITIONAL REQUIREMENT
1. Carbon
 Required by all bacteria for growth
 Autotrophs: use CO2 as the sole source of carbon
 Heterotrophs: use carbohydrates & amino acids
as source of carbon

2. Inorganic Ions:
 Phosphates, Potassium, Mg, N, S & other trace
metals
 Halophilic bacteria: require high concentration of
Sodium for optimal growth
BACTERIAL PHYSIOLOGY
NUTRITIONAL REQUIREMENT
3. Organic Nutrients:
 Carbohydrates: use by organotrophs;
 Use as energy source & initial source of carbon
skeletons for biosynthetic pathways
 Amino acids & peptone
 Vitamins

 Purines & Pyrimidines

BACTERIAL PHYSIOLOGY
NUTRITIONAL REQUIREMENT
4. Electron Donors: Ammonia, Nitrates, H2S
 Serve as sources of energy

 Litotrophic: use reduced inorganic compounds

 Heterotrophic: use reduced organic compounds

5. Electron Acceptors:
 Play a role in respiration & fermentation

 Oxygen: serves as terminal electron acceptor in

aerobic respiration
 Pyruvate, Lactate & organic compounds: products of
the terminal electron acceptance in fermentation
BACTERIAL PHYSIOLOGY
NUTRITIONAL REQUIREMENT
Oxygen
5 Groups of Bacteria based on Oxygen Requirements
1. Obligate aerobes: growth is dependent on the
availability of oxygen
2. Obligate anaerobes: growth is inhibited by the
presence of oxygen even at a low O2 tension
3. Facultative anaerobes: able to use both molecular
O2 & organic compounds as terminal electron
acceptors
4. Microaerophilic: grow best under increased CO2
tension
5. Aerotolerant: can survive but not grow for a short
period of time in the presence of atmospheric
oxygen.
BACTERIAL PHYSIOLOGY
NUTRITIONAL REQUIREMENT
Oxygen
 Tolerance to oxygen: related to the ability to detoxify
superoxide & hydrogen peroxide which are products
of aerobic respiration
 Enzymes involved in detoxification:

1. Superoxide dismutase: converts superoxide to H2O2;

present in aerobic & aerotolerant organisms
2. Catalase: converts H2O2 to water & O2; present in
all aerobic organisms except aerotolerants
 Strict anaerobes: lack both enzymes

 Superoxide: most toxic metabolites

 ENERGY METABOLISM OF
BACTERIA

Energy produced by the bacteria is produced by

 a. Fermentative Pathway

 b. Respiratory Metabolic Pathways

FERMENTATIVE METABOLISM
 Uses organic compounds as both the electron
donors & acceptors
 ENERGY METABOLISM OF
BACTERIA
3 Major Metabolic Pathways:

1. Glycolytic (Embden-Meyerhof) Pathway

 Anaerobic pathway

 Major glucose utilization pathway used by most

organisms
 Fructose-6-PO4 dehydrogenase:
 Rate limiting enzyme of the pathway
 Produces 2 moles of ATP/mole of glucose
 GLYCOLYTIC (EMBDEN-MEYERHOF)
PATHWAY
2 Phases:
a. Phase 1 Glycolysis
 Glucose is converted to Fructose-6-PO4 then to
Pyruvate
 Product: 2 moles of Pyruvate

b. Phase 2 Glycolysis
 Pyruvate formed in Phase 1 is used to oxidize
NADH
 GLYCOLYTIC (EMBDEN-MEYERHOF)
PATHWAY
Secondary Processes:
1. Homolactic fermentation
 Simplest

 Pyruvate is converted to lactate

 Utilized by Streptococci & Lactobacilli

2. Alcoholic fermentation
 Pyruvate is converted to CO2 to ethanol
 GLYCOLYTIC (EMBDEN-MEYERHOF)
PATHWAY
Secondary Processes:
3. Propionic Fermentation
 Pyruvate + CO2 forms oxaloacetate then converted to
Propionic Acid
4. Mixed Acid Fermentation
 Utilized by most members of the Family Enterobacteriaceae
 End Products: Carbon dioxide, hydrogen & ethanol

 End products of Embden Pathway: Alcohol, Gas, mixed

acids
GLYCOLYSIS
 ENERGY METABOLISM OF
BACTERIA
3 Major Metabolic Pathways:

2. Entner Duodoroff Pathway

 Aerobic pathway
 Major hexose-degrading pathway in organisms that lack
Phosphofructokinase (Pseudomonas)
 Glucose is degraded into glyceraldehyde -3-PO4 & Pyruvate
 One mole pyruvate + one mole ATP molecule formed per mole of
Glucose
 ENERGY METABOLISM OF
BACTERIA
3 Major Metabolic Pathways:

3. Warburg-Dickens Pathway
 “Pentose PO4 shunt” (Phosphogluconate Pathway)

 Generates glucose 6-PO4 which is oxidized into 6-

phosphogluconate in the presence of NADP
 Only 1 ATP generated per Glucose molecule

 Produces C4 & C5 compounds necessary for the

synthesis of different bacterial products
 RESPIRATORY METABOLISM
Aerobic Respiration:
 Most common type of respiration for human pathogens &
commensals
 Pyruvate is converted to CO2 via Acetyl CoA & TCA (Tricarboxylic
acid) cycle
 Facilitated by Electron Transport System
 Major elements of ETS:
 Cytochromes (Hemoproteins)
 Flavoproteins
 Ubiquinones (e.g Coenzyme Q
 Complete respiration: 38 molecules ATP/1 mole of Glucose
 AUTOTROPHIC METABOLISM
 Uses inorganic sources for energy
 In photosynthesis: bacteria use energy from light to
convert CO2 to Triose PO4

Anaerobic Respiration
 Bacteria use inorganic substrates (e.g Nitrogen)
as terminal acceptors instead of O2.
 BACTERIAL GROWTH
 Refers to an orderly increase in mass or
in number of all the components of cell

Measurement of Bacterial Growth:

1. In terms of Cell concentration
 Number of cells per unit volume of culture
2. In terms of Cell density
 Refers to the dry weight of cells per unit volume of
culture
 BACTERIAL GROWTH
Cells are measured through:
1. Photoelectric or turbidimetric
 Bacterial suspension scatters
light proportionally to the
number of bacteria present
 BACTERIAL GROWTH

Cells are measured through:

2. Determination of weight per unit volume
 Done through centrifugation of a known volume
of broth & determine the mass of the pellet (wet
weight)
 BACTERIAL GROWTH
Cells are measured through:
3. Microscopic counting using
Petroff-Hauser Chamber
 Principle similar to counting
Blood cells
 BACTERIAL GROWTH
Cells are measured through:

4. Electronic counting using

Coulter Counter
 Based on the negative
charge inherent to all
microorganisms
BACTERIAL GROWTH CURVE

 Bacteria divide by binary fission

 4 distinct phases of bacterial growth

a. Lag phase
b. Logarithmic (exponential) phase
c. Stationary phase
d. Decline or death phase
BACTERIAL GROWTH CURVE
 BACTERIAL GROWTH CURVE

Lag phase
 Adjustment period

 Induction of new
enzymes
 Synthesis and
assembly of ribosomes
 BACTERIAL GROWTH CURVE
Logarithmic (Exponential) Phase
 Characterized by maximal rates of cell
division
 Generation time is not similar for
different microorganisms
 Ex: 14 minutes: Pseudomonas
 24 hours: Mycobacterium
tuberculosis
 Generation time: time required for
doubling the number of bacteria
 BACTERIAL GROWTH CURVE
Stationary Phase
 Balance between cell growth
& division and cell death
 Dependent on the availability
of essential nutrients
Decline or death phase
 Bacterial cells undergo lysis