Introduction to

Bacteriology

PHARM 232
Medical Microbiology
September 11, 2020
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Outline

- Comparing prokaryotic and eukaryotic cells: an overview

- The prokaryotic cell

- Growth and nutrition of bacteria

- Bacterial growth curve

- Pathogens and opportunistic pathogens

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Learning objectives

1. Distinguish among anaerobes, aerobes, aerotolerant,

facultative anaerobes, and microaerophiles.
2. Describe what occurs at each phase of a bacterial growth
curve.
3. Explain what is meant by generation time.
4. Explain how microbial extracellular enzymes, toxins,
adhesion factors, and antiphagocytic factors affect
virulence.
5. Summarize differences between exotoxins and endotoxins.
6. Compare and contrast the terms pathogenicity, and
virulence.

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What is microbiology & why is it important?

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The good, the bad, and the nasty…

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Main differences between prokaryotic & eukaryotic cells?
Characteristics Prokaryotic Cells Eukaryotic Cells
DNA with a nuclear No Yes
membrane
Cell division Binary fission Involves mitosis
Chromosome number Usually single circular Multiple chromosomes
chromosome
Membrane-bound Absent Present
organelles, such as
mitochondria and
lysosomes
Ribosomes 70S 80S
70S in organelles
Cell wall Usually present; When present, chemically
chemically complex simple (includes cellulose
(typical bacterial cell wall and chitin)
includes peptidoglycan)

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Bacteria are prokaryotic unicellular microorganisms

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Binary Fission

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Shape and size

- Bacteria have 3 basic shapes: cocci (spherical), bacilli (rod-shaped),

and spirals.

- Some bacteria are variable in shape and are said to be pleomorphic.

- The shape of the bacterium is determined by its rigid cell wall.

- The microscopic appearance of a bacterium is one of the most

important criteria used in its identification.

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Arrangements of cocci

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Spiral bacteria

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Bacilli

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Bacterial cell components

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Cell envelope (Outer Covering)

§ Cell wall (composed of the peptidoglycan macromolecule; also known as

murein layer or mucopeptide)
§ The plasma (cytoplasmic) membrane

§ Outer membrane (gram negative only)

§ Periplasm (gram negative only)

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Structure & function of cell wall
§ Cell wall is complex structure surrounding the bacterium

§ The rigid part of the cell wall is the peptidoglycan composed of

alternating strands of N-acetylglucosamine and N-acetylmuramic acid; a
set of tetrapeptide side chains attached to the N-acetylmuramic acid; and
a set of identical peptide cross-bridges
§ Cell wall:
§ Confers protection to the cell against osmotic damage

§ Takes part in cell division

§ Is responsible for the shape of the bacterial cell

§ Possesses target site for antibiotics, lysozymes and bacteriophages

WHY ARE DRUGS THAT TARGET THE CELL WALL SYNTHESIS USEFUL?

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Cell wall structure

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Gram positive cell wall

§ In most gram-positive bacteria, the cell wall consists of many layers of

peptidoglycan layer

§ Lipoteichoic acid and wall techoic acid constitute major surface antigens

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Gram negative outer membrane and cell wall
§ Cell walls of gram-negative bacteria consist of one or a very few layers of
peptidoglycan and an outer membrane

§ The outer membrane is bilayered structure which includes the lipopolysaccharide

(LPS)
§ LPS consists of a complex lipid called lipid A, to which is attached a polysaccharide made
up of a core and a terminal series of repeat units
§ LPS is the endotoxin of gram negative bacteria – the toxicity is associated with the lipid
portion and the O polysaccharide represents a major surface antigen

§ The outer membrane has special channels known as porins, that permit the
passive diffusion of low molecular weight hydrophilic compounds. The number
and types of porins vary with bacterial species
§ Large antibiotic molecules penetrate the outer membrane relatively slowly, which
accounts for the relatively high antibiotic resistance of gram negative bacteria

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Cell wall structure

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Gram staining procedure
Bacteria with defective cell wall

§ Removal of the bacterial cell wall may be accomplished either spontaneously

or in the presence of penicillin or other agents
§ Spheroplasts: derived from gram negative bacteria

§ Protoplasts: derived from gram positive bacteria

§ L-forms – Removal of cell wall typically results in wall-less non-viable

bacteria. If such cells are able to grow and divide, they are called L forms
§ Unstable L-forms – those that revert back to their cell wall containing state when
inducing stimulus e.g. penicillin is removed
§ Stable L-forms – do not revert back to normal form

§ Mycoplasma: naturally occurring bacteria which lack cell wall e.g.

Mycoplasma pneumoniae

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Cytoplasmic membrane

§ The cytoplasmic membrane (deepest layer of the cell envelope) is a thin,

semipermeable layer which consists primarily of proteins & phospholipids

§ Functions:

1) Permeability and transport

2) Biosynthetic functions

3) Electron transport & oxidative phosphorylation

4) Chemotactic systems

Several antimicrobials exert their effects on the cytoplasmic membrane

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Cytoplasm
The cytoplasm has 2 distinct areas when seen in the electron microscope:
1) An amorphous matrix that contains ribosomes, nutrient granules, and plasmids
2) An inner, nucleoid region composed of DNA

It lacks mitochondria which is present in eukaryotic cells

Ribosomes:
They are composed of RNA & protein and organized into two subunits 30S-50S

Nucleoid:
Prokaryotes have no true nuclei; instead they package their DNA in a structure
known as the nucleoid

Several antimicrobials work by inhibiting protein synthesis on prokaryotic

ribosomes
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Cytoplasm (cont’d)

Plasmids:
Some bacteria contain extrachromosomal, double stranded, circular DNA molecules
that are capable of replicating independently of the bacterial chromosome

These confer on the bacteria certain properties such as toxigenicity and drug
resistance

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Other specialized structures

Glycocalyx (capsule or slime layer)

Functions:
1) It may play a role in the adherence of bacteria to human tissues, which is an
important step in causing infection
2) It enhances bacterial virulence

3) It plays a role in biofilm formation

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Other specialized structures (cont’d)

Flagella
§ Filamentous appendages composed of protein (flagellin) that protrudes through
the cell wall
§ They are the organs of locomotion and have characteristic patterns of distribution

Arrangements of bacterial flagella:

§ Peritrichous – numerous flagella all over the bacterial cell

§ Monotrichous – single polar flagellum

§ Lophotrichous – a tuft of flagella at one end

§ Amphitrichous – flagella at both poles of the cell

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Arrangements of bacterial flagella

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Other specialized structures (cont’d)
Pili & Fimbriae:
§ These are thin short filamentous appendages, found mainly in some gram
negative bacteria
§ Composed of structural protein subunits termed pilin

Fimbriae:
§ Adherence
§ Biofilm formation

Pili:
§ Twitching and gliding motility

§ Conjugation “sex” pili (transfer of DNA from one cell to another)

§ Adherence

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Fimbria and Flagellum

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Other specialized structures (cont’d)

Bacterial Endospores:
§ Endospores are highly resistant dormant stage of bacteria formed in unfavorable
environmental conditions such as starvation and desiccation

Endospore Formation (sporulation):

§ Endospore formation is initiated by the appearance of a clear portion of the
cytoplasm near one end of the bacterial cell which incorporates part of the
nuclear material
§ The cell membrane grows inwards forming a double layered membrane structure.
This structure is called a forespore
§ The spore wall synthesizes a thick covering layer, the cortex and a multilayered
tough outer spore coat

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Bacterial endospore formation

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Bacterial endospores
Shape and position of endospores:
§ Endospore may be central, subterminal or terminal in position
§ The diameter of the endospore may be the same as, smaller than, or larger than the
diameter of the vegetative cell

Resistance:
§ Endospores are highly resistant to ordinary boiling, heating and disinfectants
§ The highly impervious endospore coat, low water content, low metabolic activity and
high concentration of calcium dipicolinic acid make spores so much resistant to
heating and drying

Germination:
§ The process of an endospore transformation into a vegetative cell under suitable
environmental conditions
§ It occurs in 3 stages: activation, initiation, and outgrowth
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Growth and nutrition of bacteria

• Bacteria which can synthesize their essential metabolites

(organic compounds) from atmospheric carbon dioxide
and nitrogen are called "autotrophs”.
• Example: cyanobacteria

• Bacteria which cannot synthesize their own metabolites

but require preformed organic compounds for growth, are
known as "heterotrophs”.

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Factors affecting bacterial growth
Oxygen requirements

• Obligate aerobes – undergo aerobic respiration

• Examples: Pseudomonas aeruginosa (gram –ve), Bacillus (gram +ve)

• Obligate anaerobes – that cannot use oxygen in respiration and whose

growth is typically inhibited by oxygen
• Example: Clostridium

• Facultative anaerobes – able to live aerobically or anaerobically

• Examples: Staphylococci (gram +ve), Escherichia coli (gram –ve)

• Aerotolerant anaerobes – do not use aerobic metabolism but have some

enzymes that detoxify oxygen’s poisonous forms
• Example: Streptocossus mutans

• Microaerophiles – aerobes that require oxygen levels from 2 to 10% and

have a limited ability to detoxify hydrogen peroxide and superoxide radicals
• Example: Helicobacter pylori

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Oxygen requirements

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Factors affecting bacterial growth (Cont’d)

Temperature

• Effect of temperature on proteins

• Effect of temperature on lipid-containing membranes of cells and organelles

• For example, if too low, membranes become rigid and fragile

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Typical growth rates of different types of microorganisms
in response to temperature

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Factors affecting bacterial growth (Cont’d)

Hydrogen Ion Concentration (pH)

• Organisms sensitive to changes in acidity because H+ and OH- interfere with

H bonding in proteins and nucleic acids

• Neutrophiles are bacteria that grow best in a narrow range around neutral
pH (6-8)

• Acidophiles are bacteria that grow best in acidic habitats (below about pH
4)

• Alkaliphiles live in alkaline soils and water up to pH 11.5

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Factors affecting bacterial growth (Cont’d)

Water

• Microbes require water to dissolve enzymes and nutrients required in

metabolism

• Water is an important reactant in many metabolic reactions

• Most cells die in absence of water

• Some have cell walls that retain water (e.g. Mycobacterium tuberculosis)
• Endospores cease most metabolic activity in a dry environment for years

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Generation time

• Because one cell gives rise to two progeny cells, bacteria are said to undergo
exponential growth.

• The time required for a bacterial cell to divide is its generation time.

• The doubling (generation) time of bacteria ranges from as little as 20 minutes

for Escherichia coli to about 18 hours for Mycobacterium tuberculosis.

• Most bacteria have a generation time of 1-3 hours.

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Bacterial growth curve

• The doubling time varies not only with the species but also with the amount of
nutrients, the temperature, the pH and other environmental factors.

• In an enclosed vessel, referred to as a batch culture, a typical growth curve for

a population of cells is obtained.

• This growth curve can be divided into several distinct phases called lag
phase, exponential phase, stationary phase and death phase.

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Bacterial growth curve (Cont’d)
1. Lag phase
- When a microbial population is inoculated into a fresh medium, growth
usually does not begin immediately but only after a period of time called the
lag phase, which can last from few minutes up to many hours.

2. The log (logarithmic) phase [exponential phase]

- The cells begin to divide and enter a period of growth, or logarithmic increase.
- Populations in log phase are more susceptible to antimicrobial drugs.

3. The stationary phase

- This phase occurs when nutrient depletion or toxic products cause growth to
slow until the number of new cells produced balances the number of cells that
die.

4. Death Phase
- A marked decline in the number of viable bacteria.

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Bacterial growth curve

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Pathogens and opportunistic pathogens

- A microorganism is a pathogen, if it is capable of causing disease; however,

some organisms are highly pathogenic, that is, they often cause disease,
whereas others rarely cause disease.

- Opportunistic pathogens are those that rarely if ever cause disease in

immunocompetent people but can cause serious infection in
immunocompromised patients.

- Virulence is the quantitative ability of an agent to cause disease. Virulent

agents cause disease when introduced into the host in small numbers.
Virulence involves adherence, invasion and toxigenicity.

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Why do people get infectious diseases?

• People get infectious diseases when microorganisms overpower their host

defenses.

• From the organism’s perspective, the two critical determinants in

overpowering the host are the number of organisms to which the host is
exposed and the virulence of these organisms.

• From the host’s perspective, the two main arms of the host defenses are
innate immunity and acquired immunity.

• A reduction in the functioning of any component of our defenses shifts the

balance in favor of the organism and increases the chance that an infectious
disease will occur.

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Bacterial infections

• Many factors determine bacterial ability to cause infection and disease such as
attachment, toxin production, invasion, and inflammation.

Stages of bacterial
pathogenesis

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Determinants of bacterial pathogenesis

1. Transmission
Why is it important to understand the mode of transmission of bacteria and other
infectious agents?

2. Adherence to cell surface

What are the structures or substances that help bacteria to adhere to the surface
of human cells?

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Major adherence factors

Factor Example
Capsule/slime layer Pathogenic Escherichia coli – capsule
promotes adherence to the brush
border of intestinal villi

Adherence proteins Neisseria gonorrhoeae – Opa protein

on the cell binds to receptors on
epithelium

Lipoteichoic acid Streptococcus pyogenes – facilitates

binding to respiratory mucosal
receptor (along with M protein)

Fimbriae (pili) Neisseria gonorrhoeae – pili facilitates

binding to epithelium

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Adhesins

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Determinants of bacterial pathogenesis (Cont’d)
3. Invasion & intracellular survival
Several enzymes secreted by invasive bacteria play a role in pathogenesis. Among
the most prominent are:

a. Hyaluronidase - which splits hyaluronic acid, an important component of

the ground substance of connective tissue. Thus, it aids in spreading infecting
microorganisms (spreading factor).
b. Coagulase – which works in conjunction with blood factors to coagulate
plasma and leads to formation of fibrin wall around organisms.
c. Hemolysins and leukocidins - Hemolysins (dissolve red blood cells) and
leukocidins (which can destroy both leukocytes and macrophages).
d. Immunoglobulin A1 (IgA1) protease - which degrades IgA, allowing the
organism to adhere to mucous membranes, and is produced chiefly by
Neisseria gonorrhoeae, Neisseria meningitidis, Haemophilus influenzae, and
Streptococcus pneumoniae.

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Determinants of bacterial pathogenesis (Cont’d)
4. Toxin production

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Determinants of bacterial pathogenesis (Cont’d)

5. Intracellular pathogenicity
Some bacteria such as Mycobacterium tuberculosis, Brucella species and
Legionella species are able to live and grow within polymorphonuclear cells,
or macrophages.

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Important terms
Epidemic: “The occurrence in a community or region of cases of disease clearly in
excess of normal expectancy. An epidemic is relative to usual frequency of the
disease in the same area, among the specified population, at the same season of the
year”.

Pandemic: “Referring to an epidemic disease of widespread prevalence around the

globe”.

Endemic: “The constant presence of a disease or an infectious agent within a given

geographic region or population group”.

Latent disease: Some diseases result in a latent state, after which reactivation of
the growth of the organism and recurrence of symptoms may occur.

Carrier: “ A person or animal with asymptomatic infection that can be transmitted

to another susceptible host”.

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Recommended reading
Tortora et al. Microbiology: An Introduction. 12th edition. Pearson-Benjamin Cummings
Please read the following topics from Chapter 4:

- Comparing prokaryotic & eukaryotic cells: An overview

- Glycocalyx

- Fimbriae and pili

- Cell wall
- Atypical cell walls

- Damage to the cell wall (protoplasts, spheroplasts and L forms)

- The cytoplasmic membrane

- Cytoplasm

- The nucleoid
- Ribosomes

- Endospores

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Recommended reading
Tortora et al. Microbiology: An Introduction. 12th edition. Pearson-Benjamin Cummings

Chapter 6: Microbial Growth

- Oxygen
- Bacterial division & generation time
- Phases of growth
Chapter 7: The Control of Microbial Growth
- Terminology of microbial control (sterilization, disinfection, antisepsis)
Chapter 15: Microbial Mechanisms of Pathogenicity
- Adherence
- Capsules
- Enzymes
- Toxins (the difference between exotoxins and endotoxins)

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Additional references

Cappuccino,GJ., Sherman N. Microbiology: A Laboratory Manual. Eight

edition, 2008. Pearson Benjamin Cummings Publication.

Levinson, W. Review of Medical Microbiology and Immunology. Ninth

edition, 2006. McGraw-Hill Publication.

Robert W. Bauman. 2012. Microbiology with Diseases by Body System. 3rd

edition. Pearson-Benjamin Cummings Publishing Company.

Tortora G, Funke B and Case C. Microbiology: An Introduction. 12th edition.

Pearson-Benjamin Cummings.

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