MEDICAL MICROBIOLOGY I

Bacterial Anatomy

School of Nursing & Midwifery

Umma University
2023
 Introduction
 Bacteria are unicellular microorganisms
 Vary in shape ranging from 0.2-0.5 µm
 Smallest bacteria (mycoplasma) is the size of the
largest viruses (pox)
 Irrespective of the shape bacteria have common
structures e.g cell wall, cell membrane, cytoplasm,
ribosomes and the chromosome.
 Other structures such as plasmid, inclusion bodies,
capsule, fimbriae and flagella are possessed only
by some bacteria
Cell structures

EXTERNAL INTERNAL
 Capsule  Ribosomes
 Cell wall  Mesosome
 Cell membrane  Plasmids
 Bacteria Appendages  Nucleoid
 Pili/ Fimbriae  Granules
 Flagella  Spores
BACTERIAL ANATOMY
Bacterial Cell
 External structures
Capsule/Glycocalyx/Slime layer
 Structures surrounding the outside of the cell
envelope.
 A gelatinous polysaccharide or polypeptide coating
secreted by bacteria is called glycocalyx
 Capsule - glycocalyx attached firmly to the cell
wall
 Slime layer-glycocalyx attached loosely to the
cell wall.
 Glycocalyx is an extracellular polysaccharide and
helps in the formation of biofilms on inert surfaces
such as catheters, teeth and heart valves.
Capsule

 Not impermeable.
 Composed of polypeptides or polysaccharides
(exception: Bacillus anthracis (anthrax) poly-
glutamate).
 They have no net charge and hence they can’t be
stained.
 Identification made by using antiserum against the
capsular polysaccharide (Quellung reaction)
 Can also be visualized by India ink preparation or
dark-field microscopy
Capsule

 Bacteria with capsules form smooth colonies

while those without capsules form rough colonies.
 Capsules are sometimes referred as K antigens (in
Enterobacteriaceae) or as Vi antigen (in
Salmonella typhi).
 Examples of bacteria with capsules;
 Streptococcus pneumoniae, Streptococcus
mutans, Klebsiella pneumoniae, Bacillus
anthracis, Neisseria meningitides, Pseudomonas
aeruginosa, Cryptococcus neoformans
Klebsiella pneumoniae Bacillus anthracis
 Significance of capsules
 Antiphagocytic hence a virulence factor. Is
often associated with pathogenic bacteria
because it serves as a barrier against
phagocytosis by white blood cells.
 Prevent complement-mediated bacterial
cell lysis
 Growth in a biofilm prevents access of host
cells or antibiotics
 Capsules protect the cells from lysozyme.
 Permit bacteria to adhere to cell surfaces
and structures such as medical implants
and catheters.
 Significance of capsules
 Capsules can be a source of nutrients and
energy to microbes e.g Streptococcus
mutans, which colonizes teeth, ferments
the sugar in the capsule and acid
byproducts contribute to tooth decay.
 Prevent cell from drying out (desiccation)
 Capsules may protect cells from
bacteriophages.
 Attaches bacterial cell to host cell
 Capsular antigens are used as antigens in
certain vaccines e.g pneumococcal vaccine
 Cell Wall
 Found in all bacteria except mycoplasma and
metanotrophs. Mycoplasma are bacteria that have
no cell wall and therefore have no definite shape.
 Fully permeable
 Unique to bacteria hence ideal target for selective
toxicity
 Degraded by:
 Autolysins (hydrolytic enzymes produced by the
bacterial themselves) – amidases, peptidases
 Lysozymes – present in some secretions e.g. tears,
saliva, mucus
 Functions
 Maintain cell shape and rigidity – structural integrity of the cell.
The three primary shapes in bacteria are coccus (spherical),
bacillus (rod-shaped) and spirillum (spiral).
 Carries bacterial antigens– important in virulence & immunity
 Protect cell from external stresses (host)
 Contributes to virulence, promotes colonization by sticking
bacteria to surface of host cells.
 Target for antimicrobial agents (E.g. Penicillins,
Cephalosporins)
 Protects bacteria cell from osmotic lysis in hypertonic
environment.
 Protect peptidoglycan layer from action of agents such as
lysozomes
 Determines reactivity to Gram stain
 Structure of cell wall
 Made of long chains of polysaccharide
(Peptidoglycan/mucopeptide) cross-linked by short
peptides (amino acid chains).
 The polysaccharides are formed by N-Acetyl
Glucosamine (NAG) and N-Acetyl Muramic Acid
(NAM)
 The peptides contain amino acids D-alanine and
D-glutamate and a diamino acid (LL- or meso-
diaminopimelic acid, L-lysine, L-ornithine or L-
diaminobutyric acid).
 Staining the cell wall

 The cell wall stains using the Gram stain hence

can be divided into two
 Gram positive (blue)
 Gram negative (red)
Acid fast stain
 Acid fast (red on blue/Green)
 Wall-less (can’t stain)
 Spirochaetes, chlamydia & Rickettsia.
 Gram Positive cell wall

 Thick (appx 20-80 nm), multilayered

peptidoglycan layer (50-90% of cell wall material/
40+ layers of chain link fence) outside of the
cytoplasmic membrane
 Accessory polymers linked to peptidoglycan
 Teichoic acid
 Lipoteichoic acid
Teichoic acids

 Water soluble polymers of phosphate or ribitol phosphate

linked via phosphodiester bonds
 There are two types of teichoic acid, wall teichoic acid
(linked to peptidoglycan) and lipoteichoic acid (linked to
membrane)
 serve to act as chelating agents, and also for certain
types of adherence.
 Some gram positive bacteria may lack wall teichoic acid
but all contain lipoteichoic acid
 mediate attachment to mucosal membranes
 Induce septic shock in certain G+ve bacteria
 ▪Lipoteichoic acid
 Are teichoic acids made up of polyglycerol phosphate
or rubitol phosphate with sugars and amino acids
which are linked to a glycolipid in the underlying
membrane
 Induce septic shock in certain gram positive bacteria.
 Role in anchoring the cell wall to the cell membrane
 Also as an epithelial cell adhesion
Polysaccharides and proteins
 Protect peptidoglycan layer from action of agents such
as lysozymes (enzymes present in tears, mucus, saliva)
which can cleave peptidoglycan
 Sometimes promote colonization by sticking the
bacteria to the surface of host cells
 Gram Negative Cell wall
 Consist of a relatively thin single layer of
peptidoglycan (appx 10 nm) comprising 5-10% of
cell wall material.
 Do not retain the primary dye in Gram stain -
appear pink.
 Have an additional outer membrane.
 There is a periplasmic space between the inner
and outer membranes, which contains digestive
enzymes and other transport proteins.
 The outer membrane includes;
 Porin proteins: Specifically allow transport of
solutes in and out of the cell
 Lipopolysaccharides (LPS)
 Lipoprotein stabilizes the outer membrane by
anchoring it to peptidoglycan.
 The S-layer (surface layer) is directly attached to
the outer membrane, rather than the peptidoglycan
 If present, flagella have four supporting rings instead
of two because of presence of double membranes
hence a pair of rings per membrane
 Lipopolysaccharide (LPS)
 Consists of three distinct unit;
 Polysaccharide core of five
sugars,
 Phospoholipid called Lipid
A (endotoxin) responsible for
toxic effects (fever,
hypotension, septicemia)
 Outer polysaccharide of 25
repeating units- major
surface antigen (somatic or
O antigen) serving as an
antigenic determinant
 LPS also protects the cell from phagocytosis
and lysozyme.
 LPS confers a negative charge and also
repels hydrophobic molecules
 Has toxic and pyrogenic properties and is
extremely important in pathogenesis.
Differences between Gram Positive and Gram Negative Bacterial
Cells
Gram Negative Gram Positive

Has an additional cell wall layer making it bi- Has one cell wall layer
layered
Has a thin peptidoglycan layer Has a thick peptidoglycan layer

Has porins in outer membrane that act as pores Has no porin

No teichoic acids or lipoteichoic acids are Is made of teichoic acids linked to peptidoglycan
present polymers of either glycerol phosphate or rubitol
phosphate with sugars and amino acids
The S-layer is directly attached to the outer S-Layer is directly attached to the peptidoglycan
membrane, rather than the peptidoglycan and not to the membrane

If present flagella have four supporting rings If present flagella have two supporting rings
instead of two due to presence of two membrane because it has only one membrane layer
layers
Most do not sporulate Most sporulate

Does not retain the purple stain after washing off Retains the purple color even after washing off
with alcohol following staining with alcohol
 Acid-fast bacteria
 Found in Mycobacteria and Nocardia
 More complex cell envelopes than other bacteria.
 Retain stain/Acid fast-Resist decolourization with
acid-alcohol after staining with carbolfuschin
 Composed of long chained branched fatty acids
(Mycolic acid) are covalently bound via a
polysaccharide to peptidoglycan.
 Other mycolic acid containing compounds and
complex lipids form a thick waxy membranous
layer outside the peptidoglycan layer
 Cell wall-deficient cell bacteria
 Found in Mycobacteria and Nocardia
 Mollicutes- do not have a cell wall e.g
mycoplasma and ureaplasma
 L-forms or L-phase- lack cell walls derived from
bacteria that normally possess cell wall
 Result from action of:
 enzymes lytic for cell wall
 Antibiotics inhibiting peptidoglycan biosynthesis
 Protoplasts - Have their cell wall entirely removed
 Spheroplasts - Have their cell wall only partially
removed
 Cytoplasmic Membrane

 Just inside the peptidoglycan layer of the cell wall

lies the cytoplasmic membrane, which is
composed of a phospholipid bilayer similar in
microscopic appearance to that in eukaryotic cells.
 They are chemically similar, but eukaryotic
membranes contain sterols, whereas prokaryotes
generally do not.
 The only prokaryotes that have sterols in their
membranes are members of the genus
Mycoplasma.
 Functions of Cell Membrane

1) Barrier-selectively permeability determines what

molecules pass through membrane
a) large(protein, starch, etc.) don’t pass through
easily
b) small (H2O, O2, CO2) pass through easily
2) Secrete exoenzymes that breakdown nutrients to
provide energy
a) Examples-amylase & lipase
3) Cell wall synthesis enzymes located here
 Cytoplasm

 Substance located within cell membrane

– 80% water (other-protein, carbs, lipids,
inorganic ions)
– Thick, elastic, semitransparent
– Contains DNA and ribosomes
 Flagella
 Long (3 to 12 µm), filamentous surface appendages
 Originate from the basal body in the cytoplasm .
 Responsible for bacterial motility and chemotaxis by
rotating in a propeller- like manner by means of a "motor"
located just under the cytoplasmic membrane.
 Are composed of flagellins (proteins)
 They are anchored to the plasma membrane and protrude
through the cell wall to the external part of the cell.
 Posesed by most rods/bacilli
 Presence shown by motility test e.g. hanging drop
preparation
 Flagella - Arrangements
1. Monotrichous- a single
flagellum at one end of the cell
2. Lophotrichous- a tuft of
flagella (2 or more) at one end
of the cell
3. Amphitrichous- one flagella
at the opposite ends of the cell
4. Peritrichous- flagella present
around the entire body of the
cell.
5. Amphilophotrichous –
Clusters of flagella at both
ends
 Functions of flagella
 Virulence factors -allow bacteria to evade
phagocytes by swimming away from them
 Allow bacterial cell to come into close contact with
the adhesion receptors on the human cell.
 Source of H-antigen used in serotyping motile
spp. of bacteria.
 Serve as antigenic determinants.
 Recognized by TLR5
 Pilli
 Hollow, Hair-like projections on the surface of the cell
 Shorter, Straighter and thinner than flagella
 Composed of subunits of a protein (pilin)
 Mostly on gram negative bacteria
 Sex pili:
 Relatively long but is often found in few no. (1 to 6)
 Involved in conjugation i.e transfer of DNA between
bacteria
 Receptor for bacteriophages.
 Common pili (Fimbriae)
 Relatively short and many (about 200).
 Provide means for attachment/ adhesions and
colonization ) of gram negative bacteria to host cells
Salmonella
Internal structures
Cytoplasmic Components
 Bacteria have a very simple internal structure,
and no membrane-bound organelles.
 Cytoplasm - Forms the site for all cellular
metabolic processes
 Mesosome
 Ribosomes
 Nucleoid
 Nutrient granules
Mesosomes

 Appear as convoluted indentations in the

cytoplasmic membrane
 More prominent in Gram +ve bacteria
Functions
 Are sites of respiratory enzyme activity
 Coordinate nuclear & cytoplasmic division during
binary fission
 Phagocytosis
Ribosomes
 They are composed of RNA and proteins.
 Site of activity of antimicrobials that disrupt protein
synthesis e.g aminoglycosides, tetracyclines,
chloramphenicol, macrolides – erythromycin
 Ribosomes give the cytoplasm of bacteria a granular
appearance in electron micrographs. Though smaller than
the ribosomes in eukaryotic cells, these inclusions have a
similar function in translating the genetic message in
messenger RNA into the production of peptide sequences
(proteins).
 70S in size with 50S and 30S subunits
 50S subunit contains the 23S and 5S rRNA
 30S contains the 16S rRNA
 Function; Site of protein synthesis
Nucleoid
 This
is the location of DNA in cytoplasm and
has homogenous material.
 Bacterial
DNA consists of a single, circular
double-stranded DNA
 Lacksnuclear membrane (called nucleoid or
nuclear body)
 Thenucleoid also lacks nucleolus, histones,
and mitotic spindle
 Contains genetic material that codes for all
genetic information expressed by the cell
 Nutrient granules/Storage granules
 Reserveof energy and phosphate for cell
metabolism. Nutrients may be stored in the
cytoplasm in the form of glycogen, lipids,
polyphosphate, or in some cases, sulfur or nitrogen.
 Inclusion
granules are composed of volutin, lipid
and polysaccharide.
 Staincharacteristically with certain dyes e.g volutin
stains red with methylene blue.
 Examples; metachromatic granules in diphtheria
bacilli
 Function; Serve as storage area for nutrients and
energy for cell metabolism
Additional Organelles

 Spores
 Plasmids
Spores
 It may be round, oval, or elongated
 Formed inside the parent cell
(Endospores) and contains bacterial DNA
 Formed in response to nutrient depletion,
extreme environments or stress
 They do not multiple to increase in number
because one cell yields just one spore
 Composed of a thick keratinlike coat
responsible for resistance
 Contain dipicolinic acid
Spores
 Resistantto heat, chemicals, radiation and drying and
can remain dormant for hundreds of years
 Formed by bacteria like Clostridia, Bacillus
 Some bacteria of Clostridia and Bacillus species e.g
Clostridium botulinum, form spores that are highly
resistant to drying, high temperature, radiation and
other environmental hazards.(need autoclave at
121˚C, 15 min) Once the hazard is removed, the
spore germinates to create a new population. Hence
the need to bury infected organisms even deeper into
the soil
 The spore may be positioned at the end (terminal) of
the bacterium or centrally (median).
Shape & position of bacterial spore
Oval central

Spherical central

Oval sub terminal

Oval terminal

Spherical terminal
Endospores
 When bacteria are generally subjected to
increasingly unfavourable environmental
conditions, including lack of water, depletion of
nutrients and deviation of temperature from
optimum, which are suitable for growth and
multiplication, they may die. But certain rod-
shaped bacteria under the family Bacillaceae
are able to develop structures called
endospores i.e. spores within cells.
 These spores are formed to enable the
bacteria withstand the unfavourable
conditions.
 Bacterial cells without spores are called
vegetative cells.
Not all spores are equally resistant to adverse
physico-chemical forces. Spores of the common
soil bacterium Bacillus subtilis can withstand 100 oC
for 3 hours while spores of Clostridium botulinum
can withstand the same temperature for 4 hours.
 Spores of thermophilic bacteria such as Bacillus
stearothermophilus are more resistant to heat
whereas non-sporing bacteria are killed by
exposure to temperatures of 65-80 oC for only a few
minutes.
 Spores can remain dormant and stable for many
years, and have been cultured from spores 300
years old. When spores are exposed to favourable
conditions, they germinate into vegetative cells.
Bacillus anthracis
 Plasmids
 Small circular, self-replicating, double-stranded
circular DNA molecules and exist independently of
chromosomal DNA in bacterial cells.
 Are extrachromosomal hence can replicate
independently during cell division and may be
inherited by both daughter cells.
 Transferred between bacteria by conjugation through
sex pili
 Used in genetic engineering
 Free or integrated into the chromosome
 Transmitted to daughter cells during binary fission
 Confer properties like resistance and virulence factors
 Classes of plasmids

1. Transfer and sex factors (plasmids)

 They are conjugative plasmids which determine
their own transfer in bacteria and also promote
transfer of other genes from donor to recipient
cells during conjugation. Some conjugative
plasmids can cause the transfer of chromosomal
genes during conjugation. Such plasmids are
termed as sex factors.
2. Resistance (R) plasmids
 These are plasmids responsible for infectious
antibiotic resistance. The “R” factors plasmids
carry genes that encode for enzymes responsible
resistance to various antibiotics.
3. Virulence / pathogenicity plasmids

 These are plasmids that carry genes which

encode for toxigenic and invasive properties e.g. -
(i) Enteric plasmids which encode for heat stable
(ST), and both heat stable (ST) and heat liable
(LT) toxins in E . coli
4. Bacteriocins/col. plasmids
 These are plasmids commonly found in E . coli
which encode bacteriocins (colicins). Some of
these plasmids are conjugative and others non-
conjugative.
5. Staphylococcal plasmids.
 These are plasmids found in S . aureus which
encode for antibiotic and heavy metal resistance,
and production of enterotoxins and haemolysis.
They are non-conjugative and specific to
Staphylococcus.
6. Mercury resistance plasmids
 They are plasmid common in some strains of enteric
bacteria, Pseudomonas and S. aureus which
enable the host tolerate otherwise toxic
concentrations of mercury and other heavy metal
ions. They also specify the conversion of relatively
harmless mercury compounds to neurotoxic
organomercurial methyl compounds.
7. Degradative plasmids

 They enclose for versatile enzymes which enable

the host to degrade many organic molecules,
including complex organic compounds including
xenobiotics which are reculcitrant. The versatility
of pseudomonas is due to the presence of
plasmids which code for enzymes of catabolic
pathways of a number of complex organic
substrates.
8. Tumour inducing plasmids
 A large plasmid has been found in the bacterium
Agrobacterium tumefaciens which causes crown gall
disease in plants. Avirulent strains do not have the
plasmid.
9.Cryptic plasmids
 They are small plasmids found in many genera of
bacteria, for which no biological role has been
established.
 Transposons (Tn elements)
 Many plasmid genes determining drug resistance
are known to be located in discrete DNA units.
These genes may be dissociated from the plasmids
and be transposed into another DNA molecule, and
from there undergo further translocation.
 Thus a gene can hop from a drug resistance plasmid
to the chromosome, to another plasmid, or to a
temperate phage (prophage, provirus) genome.
Such movable units of DNA have been called
transposons, transposable elements translocatable
elements, drug resistance determinants or jumping
genes.