Antibiotics 1

Antibiotics
Lecture 1: Classification of Bacteria

Learning Objectives
Be able to classify common bacteria by gram
stain, morphology, and other discriminating
characteristics.
Bacterial Names
Most pathologic bacterial species are colloquially referred to by genus

name only (e.g. Klebsiella, Proteus, Salmonella, Pseudomonas).
Some are referred to by specific species name (e.g. E. coli).
Some are referred to by genus name only, but probably should be referred
to more specifically (e.g. Enterococcus, “Staph”, and “Strep”).
Classification of Bacteria
Morphology (cocci vs. rod/“bacilli” vs. coccobacilli)
Gram Stain (positive vs. negative vs. variable)
Growth Requirements (aerobic vs. anaerobic)
Biochemical Reactions (lactose fermenting vs. non-fermenting)
Serotype (group A vs. B vs. D streptococcus)
Antibiotic Resistance Patterns (MSSA vs. MRSA)
rRNA Sequence Analysis
Bacterial Morphology
Along with gram staining, general morphology are the two

most important methods of differentiating bacteria.
Coccus / Cocci Rod / Rods

Gram Stain
Different types of bacteria have different cell wall structures.
“Gram Positive” Bacteria “Gram Negative” Bacteria

Outer Membrane
(lipopolysaccharide & protein)
Peptidoglycan
(polymer of sugars & amino acids)
Plasma Membrane
(phospholipid bilayer)
Gram staining uses these differences to create distinguishing colors

after application of a sequence of stains.
Gram Stain
Application of crystal violet Application of iodine Alcohol wash Application of safranin

Gram Stain
Staphylcoccus
Clostridium
Streptococcus Actinomyces
Enterococcus
Peptostreptococcus
E.coli, Proteus, Klebsiella,
Serratia, Enterobacter,
Neisseria Shigella, Salmonella,
Moraxella Yersinia, Pseudomonas,
Stenotrophomonas,
Bacteroides
Gram Stain
Coccobacilli
Gram + Listeria
Haemophilus
Gram - Legionella
Bordetella
Gram Acinetobacter
Variable (sometimes listed as
rods, or pleomorphic)
Additional Clinically Important Tests
Coagulase test
Hemolysis test
Lancefield serotyping
Lactose Fermentation
Coagulase Test
Coagulase is an enzyme produced by some bacteria that converts

fibrinogen to fibrin.
The presence of coagulase is used predominantly to differentiate

Staphylococcus spp.
Coagulase Positive Coagulase Negative

S. aureus Most other Staph. spp.
Some S. intermedius
(e.g. S. epidermidis, S. capitis,
S. lugdunensis, S. saprophyticus)
Hemolysis Test
Streptococcal species are further classified based on their ability to

hemolyze red blood cells.
Usually tested by culturing known Streptococcal bacteria on blood

agar.
Hemolysis Test
Blood Agar
Alpha Hemolytic Beta Hemolytic Gamma Hemolytic

(“Partial Hemolysis”) (Complete Hemolysis) (No Hemolysis)
S. pneumoniae S. pyogenes S. bovis

Viridans strep. S. agalactiae Enterococcus
Lancefield Serotypes
Streptococcus/Enterococcus are sometimes grouped according to

specific carbohydrates present on the bacterial wall.
Group A – S. pyogenes
Beta hemolytic
Group B – S. agalactiae
Group D – Enterococcus, S. bovis Gamma hemolytic
No Lancefield antigens – S. pneumoniae,

Alpha hemolytic
Viridans strep
Ability to ferment lactose is a major differentiating characteristic of gram

negative rods.
Usually tested by culturing unknown bacteria on MacConkey agar.
Component Function
Agar Structural support for growth
Bile Salts Inhibits growth of gram positive bacteria
Crystal Violet Inhibits growth of gram positive bacteria
Lactose Food for lactose-fermenting bacteria  Results in ↓ pH
Peptides Food for non-lactose fermenting bacteria  Results in ↑ pH
Neutral red Dye which changes to a red color at low pH
MacConkey Agar
Lactose Fermenters Lactose Non-Fermenters

(i.e. “Lactose positive”) (i.e. “Lactose negative”) Lactose Slow Fermenters
E. coli, Pseudomonas, Citrobacter,

Klebsiella, Proteus, H. Flu, Serratia
Enterobacter Salmonella, Shigella,
Stenotrophomonas,
Acinetobacter
A Word About Enterococcus…
Enterococcus, formerly known as group D streptococcus,

became a separate genus in the 1980s.
Some micro labs don’t differentiate Enterococcus species...
Species Prevalence Trend in Antibiotic Resistance

E. faecalis Majority of enterococcal Relatively less resistant to
infections ampicillin, vancomycin and
aminoglycosides
E. faecium Sizable minority of Relatively more resistant to
enterococcal infections, amipicillin, vancomycin and
particularly nosocomial aminoglycosides
“Anaerobes”
Anaerobes include all bacteria which grow and reproduce only in the
absence of oxygen.
They are predominantly found in the GI tract (including the oral cavity).
While they are generally stainable via gram stain, they are rarely referred
to as gram positive or negative in clinical practice.
Examples: Clostridium
Bacteroides
Peptostreptococcus
Actinomyces
“Atypicals”
An inexact term applied to bacteria which are particularly unusual in

cellular structure, morphology, biochemistry, or life-cycle.
Examples: Mycoplasma (lack a cell wall, highly pleomorphic)
Chlamydia (lack peptidoglycan in the cell wall,

multiphased life-cycle, obligate intracellular parasites)
Rickettsia (obligate intracellular parasites, generally

require arthropod vectors)
Legionella (does not grow on traditional media)

A Practical Classification of Bacteria
Gram Positive Gram Negative Anaerobic “Atypical”
Cocci Coccobacilli Cocci Rod Coccobacilli Clostridium Mycoplasma

Bacteroides Chlamydia
Peptostreptococcus Rickettsia
Actinomyces Legionella
Listeria Neisseria
Moraxella
Clusters Pairs, Chains Haemophilus

Bordetella
Lactose Lactose Non- Lactose Slow
Streptococci Fermenting Fermenting Fermenting
Staphylcocci Enterococci Gram Variable,
Coccobacilli
E. coli Pseudomonas Serratia
Coagulase Test Hemolysis Test Klebsiella Proteus Citrobacter Acinetobacter
Lancefield Serotyping Enterobacter Stenotrophomonas
More Bacteria Shorthand
Streptococcus pneumoniae – “Pneumococcus”
Haemophilus influenzae – “H. flu”
Enterobacteriaceae (e.g. E. coli, K. pneumoniae, P. mirabilis) –

“Enterics”, “Enteric gram negative rods”
Clostridium difficile – “C. diff”
Any and all anaerobic species – “Anaerobes”

More Bacteria Shorthand
Methicillin-sensitive Staph. aureus – MSSA
Methicillin-resistant Staph. aureus – MRSA
Vancomycin-resistant Enterococcus – VRE
Extended-spectrum beta-lactamase producing member of the

Enterobacteriaceae family – ESBL
Music: Invention #13 in A minor, BWV 784
by Johann Sebastian Bach
Original arrangement for 2 guitars

Rendered in Finale 2012
Sound provided by Garritan
Copyright © 2013 by Eric Strong
This work is made available under the terms of the Creative Commons
Attribution-NonCommerical-NoDerivs 3.0 Unported License
For details, please refer to: creativecommons.org/licenses/by-nc-nd/3.0/deed.en_US

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Antibiotics

Lecture 1: Classification of Bacteria

Most pathologic bacterial species are colloquially referred to by genus

Some are referred to by specific species name (e.g. E. coli).

Along with gram staining, general morphology are the two

Coccus / Cocci Rod / Rods

“Gram Positive” Bacteria “Gram Negative” Bacteria

Gram staining uses these differences to create distinguishing colors

Application of crystal violet Application of iodine Alcohol wash Application of safranin

Coagulase is an enzyme produced by some bacteria that converts

The presence of coagulase is used predominantly to differentiate

Coagulase Positive Coagulase Negative

Streptococcal species are further classified based on their ability to

Usually tested by culturing known Streptococcal bacteria on blood

Alpha Hemolytic Beta Hemolytic Gamma Hemolytic

S. pneumoniae S. pyogenes S. bovis

Streptococcus/Enterococcus are sometimes grouped according to

Group D – Enterococcus, S. bovis Gamma hemolytic

No Lancefield antigens – S. pneumoniae,

Ability to ferment lactose is a major differentiating characteristic of gram

Usually tested by culturing unknown bacteria on MacConkey agar.

Lactose Fermenters Lactose Non-Fermenters

E. coli, Pseudomonas, Citrobacter,

Enterococcus, formerly known as group D streptococcus,

Some micro labs don’t differentiate Enterococcus species...

Species Prevalence Trend in Antibiotic Resistance

An inexact term applied to bacteria which are particularly unusual in

Examples: Mycoplasma (lack a cell wall, highly pleomorphic)

Chlamydia (lack peptidoglycan in the cell wall,

Rickettsia (obligate intracellular parasites, generally

Legionella (does not grow on traditional media)

Gram Positive Gram Negative Anaerobic “Atypical”

Cocci Coccobacilli Cocci Rod Coccobacilli Clostridium Mycoplasma

Clusters Pairs, Chains Haemophilus

Streptococcus pneumoniae – “Pneumococcus”

Haemophilus influenzae – “H. flu”

Enterobacteriaceae (e.g. E. coli, K. pneumoniae, P. mirabilis) –

Clostridium difficile – “C. diff”

Any and all anaerobic species – “Anaerobes”

Methicillin-sensitive Staph. aureus – MSSA

Methicillin-resistant Staph. aureus – MRSA

Vancomycin-resistant Enterococcus – VRE

Extended-spectrum beta-lactamase producing member of the

Original arrangement for 2 guitars

Copyright © 2013 by Eric Strong

For details, please refer to: creativecommons.org/licenses/by-nc-nd/3.0/deed.en_US

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