A.
OVERVIEW OF THE MICROBIAL WORLD
B. CLASSIFICATION/TAXONOMY
C. COMPARISON OF PROKARYOTIC & EUKARYOTIC CELL STRUCTURE
D. BACTERIAL MORPHOLOGY
OBJECTIVES
o Describe microbial classification (taxonomy) & accurately apply the
rules of scientific nomenclature for bacterial names
o Differentiate between prokaryotic & eukaryotic cell types
o Differentiate the cell walls of gram-positive from gram-negative
bacteria
INTRODUCTION TO MICROBIOLOGY
Microbiology
Study of living organisms of microscopic size (microorganisms)
Medical Microbiology
Subdivision of microbiology concerned with the causative agents
of infectious diseases of man, the response of the hosts to
infection & various methods of diagnosis, treatment &
prevention
Microscopy
Most common method used both for the detection of
microorganisms directly in clinical specimens & for the
characterization of organisms grown in culture
Defined as the use of a microscope to magnify objects too small
to be visualized with the naked eye so that their characteristics
are readily observable
MICROBIOLOGY HISTORY
st
Antonie van Leeuwenhoek – 1 person to describe
microorganisms
Louis Pasteur – “Father of Microbiology”
Robert Koch: Koch’s postulates are used to prove a direct
relationship between a suspected pathogen & a disease
MICROORGANISMS
Large & diverse group of microscopic organisms that exist as
single cells or cell clusters
Includes viruses, which are microscopic but not cellular
Includes prions, viruses, bacteria, fungi & parasites
Naming Microorganisms
In a nomenclature system designed by Carolus Linnaeus (1735),
each living organism is assigned two names
The two names consists of a genus & a specific epithet, both of
which are underline or italicized
Prions
Infectious protein capable of causing chronic neurologic diseases
Virus
Acellular entities that must invade a host cell in order to replicate
Smallest of all microbes
Consist of a nucleic acid core (DNA or RNA) surrounded by a
protein coat
Diverse group of microorganisms that range from unicellular
forms (yeasts) to molds & mushrooms
Absorb nutrients from their environment including the organic
molecules that they use as a source of carbon & energy
Parasites
Organism that lives in or on another organism (its host) and
benefits by deriving nutrients at the host's expense
Bacteria
Unicellular organisms that lack a nuclear membrane & true
nucleus
Classified as prokaryotes (having no mitochondria, endoplasmic
reticulum or golgi bodies)
Absence of the preceding bacterial cell structures differentiate
them from eukaryotes
Characteristics of Bacteria
Prokaryotic
Has both RNA & DNA & multiplies by binary fission
PROKARYOTES VS. EUKARYOTES
Prokaryotic & Eukaryotic cells are similar in their chemical
composition & chemical reactions
It is primarily the structures of cell walls & membranes & the
absence of organelles (specialized cellular structures that have
specific functions) that distinguish prokaryotes from eukaryotes
Chief distinguishing characteristics of Prokaryotes
Their DNA is not enclosed within a membrane & is usually a
singular circularly arranged chromosome
Their DNA is not associated with histones; Other proteins are
associated with the DNA
They lack membrane-enclosed organelles
Their cell walls almost always contain the complex
polysaccharide peptidoglycan
They usually divide by binary fission. During this process, the
DNA is copied & the cell splits into two cells. Binary fission
involves fewer structures & processes than eukaryotic cell
division
Chief distinguishing characteristics of Eukaryotes
Their DNA is found in the cell’s nucleus which is separated from
the cytoplasm by a nuclear membrane & the DNA is found in
multiple chromosomes
Their DNA is consistently associated with chromosomal proteins
called histone & with nonhistones
They have a number of membrane-enclosed organelles including
mitochondria, endoplasmic reticulum, golgi apparatus,
lysosomes & sometimes chloroplasts
Their cells walls when present are chemically simple
Cell division usually involves mitosis in which chromosomes
replicated & an identical set is distributed into each of two
nuclei. Division of the cytoplasm & other organelles follows so
that the two cells produced are identical to each other
 ☺
An envelope may surround the coat
BACTERIAL CLASSIFICATION, MORPHOLOGY & ULTRASTRUCTURES Microbiology

Ends may be square or rounded Can

CHARACTERISTIC PROKARYOTES EUKARYOTES have tapered, pointed ends &
TYPICAL SIZE 0.2 – 2.0 m 10 – 100 m
NUCLEUS No nuclear ☺ are termed as fusiform
True nucleus consisting of
membrane or nuclear membrane & nucleoli Spirochetes
nucleoli Spiral in form
MEMBRANE- Absent Present; examples include
ENCLOSED lysosomes, golgi apparatus, Vary in length & in the number of helical
ORGANELLES endoplasmic reticulum, turns (not all helical bacteria are
mitochondria & choloroplasts called spirochetes)
GENETIC Nucleoid Nucleus
MATERIAL
EXAMPLE Bacteria Human Cell, Fungi, Parasites,
Animal Cell
SIZE OF NUCLEUS 70s with 50s & 80S with 60S & 40s subunits
30s subunits
REPRODUCTION Asexual (Binary Sexual & Asexual
Fission)
CELL WALL Present except ☺
Some have cell wall
Mycoplasma

BACTERIAL MORPHOLOGY

Pleomorphic

When bacterial specifies varies in size & shape within a pure culture

THE GRAM STAIN

Significance of Staining:
Cocci
From kokkos meaning berry Impart color or fluorescence needed to visualize bacteria under the
microscope
Spherical or nearly spherical
Gram Staining
May occur singly, in pairs (diplococci) or
in clusters (staphylococci) Most common used stain in the clinical microbiology laboratory
Provides a mechanisms form the rapid presumptive
Bacilli
From baculus meaning rod identification of pathogens

Relatively straight, rod-shaped Gives important clues related to the quality of a specimen &
(cylindrical) cells whether bacterial pathogens from a specific body site are
considered normal flora colonizing the site or the actual cause
of infection
May vary greatly in size & length from
very short coccobacilli to long
filamentous rods First devised by Hans Christian Gram

Used to divide most bacterial speciies into two large groups:

 Spirochetes (gram negative cell wall but are too small to be seen
◌ Gram-positive (blue to purple) with light microscopy; hard to stain because it is very thin)
◌ Gram-negative (pink)

Cell wall of the bacteria determines the gram-staining characteristics

of a species PARTS OF BACTERIAL CELL
REAGENT PURPOSE GRAM (+) GRAM (-)
1. Cell Envelope
CRYSTAL Primary Stain Stain Purple Stain Purple
VIOLET
2. Cellular Appendages
GRAM’S Mordant Remains Remains
IODINE Purple Purple
95% Decolorizer Remains Becomes 3. Cytoplasmic Structures
ALCOHOL OR Purple Colorless
ACETONE
SAFRANIN Counterstain Remain Stain Pink
Purple
Capsule
Often referred to as the “slime layer” if not well-defined or

“glycogcalyx” if well-defined capsule

Rules in Gram Staining
All encapsulated microorganism are made up of polysaccharide gel
1. All COCCI are Gram Positive EXCEPT Bacillus anthracis Polypeptide / Poly D glutamic acid
except: Neisseria
Serve either to inhibit phagocytosis or to aid in adherence to host tissue
Veilonella or synthetic implants (virulence factor), immunogenicity
Moraxella
Also a cellular appendage
2. All Bacilli are Gram Negative
except: Mycobacterium Seen in both gram positive & negative bacteria

Corynebacterium ENCAPSULATED ORGANISMS

Actinomyces SOME Streptococcus pneumoniae
Streptomyces KILLERS Klebsiella pneumoniae
Bacillus HAVE Haemophilus influenza
PRETTY Pseudomonas aeruginosa
Clostridium NICE Neisseria meningitides
Erysipelothrix CAPSULE Cryptococcus neoformans (only fungi with capsule)
Listeria
Other Encapsulated organisms (BES)
Nocardia 1. Bacillus anthracis & Bordatella pertussis
Bacterium (Bifido-, Proprioni-, Eu-, Arcano-)
2. Enterobacter aerogenes & Escherichia coli (meningeal strains)

3. Not Gram Stained are:

3. Streptococcus pyogenes
Rickettsiae (intracellular)
4. Salmonella typhi (virulence) virulence antigen
Mycoplasma & Ureaplasma (No Cell Wall)
BACTERIAL CLASSIFICATION, MORPHOLOGY & ULTRASTRUCTURES
CELL ENVELOPE
Outer Membrane
Contains proteins, phospholipids & lipopolysaccharides (LPS)
◌ LPS contains three regions:
□ Antigenic O
□ Core polysaccharide
□ Inner lipid A (endotoxin)
Responsible for producing fever & shock conditions in patients
infected with gram-negative bacteria
Functions In the following ways:
◌ Acts as a barrier to hydrophobic compounds & harmful
substances – “major permeability barrier’
◌ Provides attachments sites that enhance attachment to
host cells
Only seen in gram-negative bacteria
Microbiology
Cell Wall
Often referred to as the peptidoglycan or murein layer
Structure is unique & composed of alternating sugar components
(N-acetyl-d-glucosamine [NAG] & N-acetyl-d-muramic acid
[NAM])
◌ Polymers of these subunits cross-link to one another
forming a multilayered, cross-linked structure of
considerable strength
Gives the bacteria cell shape & strength to withstand changes in
environmental osmotic pressures that would otherwise result in
cell lysis
Usually the target of antibiotics & enzymes
Found in all free living bacteria whether gram-positive or
gram-negative except mycoplasma & ureaplasma (no cell wall)
☺
Basis of gram staining
Gram-Positive VS. Gram-Negative Cell Wall
Gram-Positive Bacteria
◌ Thicker peptidoglycan layer (85% of cell wall)
◌ Contains teichoic acid for attachment to host
cell Gram-Negative Bacteria
◌ Thinner Peptidoglycan layer (15% of cell wall)
Acid-Fast Cell Wall
Certain bacteria have a gram-positive cell wall structure but also
contain a waxy layer of glycolipids & fatty acids (mycolic acid)
bound to the exterior of the cell wall
Phospholipid bilayer with embedded proteins that envelop the
cytoplasm (hydrophobic “cell sack”
Location of the ETC where energy is generated
Transport of solutes into & out of the cell

More than 60% of the cell is lipid making it resistant to drying & Seen in both gram-positive & gram-negative bacteria
chemicals making it difficult to stain with the Gram stain

Best stained with an acid-fast stain in which the bacteria are stained
with carbolfuchsin, followed by acid-alcohol as a decolorizer

Other bacteria are decolorized by acid-alcohol but they retain the

stain making them designated as acid-fast bacterias

Acid Fast Bacterias include:

1. Crytosporium sp. 4. Mycobacterium sp.

2. Legionella micdadei5. Nocardia sp. (partially acid-fast)

3. Isospora sp.

GRAM-POSITIVE CELLS GRAM-NEGATIVE CELLS

WITH 2 LAYERS: WITH 3 LAYERS:
Periplasmic Space 1. Inner Cytoplasmic 1. Inner Cytoplasmic
Membrane Membrane
Bounded by the internal surface of the outer membrane & the 2. Outer THICK 2. THIN Peptidoglycan layer
external surface of the cell wall Peptidoglycan Layer 3. Outer Membrane with
LPS
Consists of gel-like substances containing nutrient-binding proteins Low Lipid Content High Lipid Content
NO Endotoxin (+) Endotoxin (LPS) – Lipid A
Also contains several enzymes involved in the degradation of NO Periplasmic Space (+) Periplasmic Space
macromolecules (like - lactamases) & detoxification of (+) Teichoic Acid NO Teichoic Acid
environmental solutes Vulnerable to antibiotics & Resistant to antibiotics &
enzymes enzymes
Only seen in gram-negative bacteria Protoplast Spheroplast

Why is Gram-positive bacteria more vulnerable to antibiotics?

Cytoplasmic Membrane Because it has a thicker cell wall which is the target of antibiotics
BACTERIAL CLASSIFICATION, MORPHOLOGY & ULTRASTRUCTURES Microbiology

5. Peritrichous – flagella that occurs on

Protoplast all sides of bacteria

Bacteria who have their cell wall entirely removed

Derived from gram-positive bacteria

Axial Filaments
Seen in spirochetes (internal flagellum)
Spheroplast
Bundles of fibrils that are used by spirochetes to move
Bacteria who have their cell wall only PARTIALLY removed
Derived from gram-negative bacteria Rotation of the filaments produces a movement of the outer sheath
that propels the spirochetes in a spiral motion similar to the
way a corkscrew moves through a cork

CELLULAR APPENDAGES

CYTOPLASMIC STRUCTURES
Fimbriae or Pili
Includes Nucleoid, Plasmids, Ribosomes, Cytoplasmic Granule &
Hairlike, proteinaceous structures that extend from the cell Endospores
membrane into the external environment
All cytoplasmic structures are found in both gram-positive &
Consist of a protein called pilin gram-negative bacteria EXCEPT endospore (gram—positive
Three types: only)

◌ Common (Ordinary) Pili – used for the attachment of the

bacteria to host cell
◌ Sex (Conjugation) Pili – used to attach bacteria to another
bacterial cell
◌ Virulence Pili – for the production of
infection/pathogenicity (observed with N. meningitides)
Only seen in gram-negative bacteria
Nucleoid
AKA Nuclear Body, Chromatin Body, Nuclear Region
DNA in the bacterial cell is generally in the central region
Unlike in eukaryotes, it is not enclosed within a membrane-bound
nucleus

Highly coiled DNA is intermixed with RNA & various proteins that
lend structural support
Flagella
Organ of locomotion

Exterior protein filaments that rotate & cause bacteria to be motile

Number and arrangement of flagella are sometimes used for

identification purposes

Seen in both gram-positive & gram-negative bacteria

Classification of bacteria according to flagella

1. Atrichous – no flagella

2. Monotrichous – single flagellum at

one end

3. Lophotrichous – tuft of flagella at

one or both ends

4. Amphitrichous – single flagellum at

both ends
Plasmids
Extrachromosomal DNA Endospores

Non-essential genetic material that are not connected to the main Some bacteria produce endospores in response to hard,
bacterial chromosome environmental conditions

Generally not crucial for the survival of bacterium under normal
conditions BUT under certain conditions, plasmids are an
advantage to cells Plasmids may carry genes for such activities
as antibiotic resistance, production of toxins & synthesis of
enzymes
Resistant structure enabling bacteria to withstand adverse
conditions (exposure to chemical agents, temperature changes,
starvation, dehydration, UV & gamma radiations & desiccation
Highly resistant because of calcium dipicolinate/dipicolonic acid

Transferred through conjugation Bacteria with spores:

1. Bacillus sp.
2. Clostridium sp.
3. Coxiella burnetti
Ribosomes

Function as sites of protein synthesis Called

70S ribosomes with two subunits
Summary
◌ 50S subunit
Structures Gram-Positive Gram-Negative
Bacteria Bacteria
◌ 30S subunit
Capsule (+) (+)
Outer Membrane (-) (+)
with LPS
Cell Wall Thick Thin
Cytoplasmic Granules Periplasmic Space (-) (+)
Function as storage deposits (accumulated food reserves)
Cytoplasmic (+) (+)
Membrane
Consists of polysaccharides such as glycogen, lipids &
polyphosphates Fimbriae or Pili (-) (+)
Flagella (+) (+)
Examples: Nucleoid (+) (+)
1. Corynebacterium Babes-Ernst Granules Plasmids (+) (+)
Ribosomes (+) (+)
2. Mycobacterium Much Granules Cytoplasmic (+) (+)
Granules
3. Yersinia pestis Volutin granules Endospores (+) (-)
Axial Filaments Spirochetes ONLY