MICROBIOLOGY

014-1-MI
INTRODUCTION TO THE BACTERIAL CELL

NAME: STUDENT NO. _

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MICROBIOLOGY
014-1-MI
INTRODUCTION TO THE BACTERIAL CELL

TABLE OF CONTENTS

PAGE

OBJECTIVES 2

RESOURCE MATERIAL 4

OVERVIEW OF THE CELL 5

THE OUTSIDE OF THE BACTERIAL CELL 7
FLAGELLA 7
FIMBRIAE AND PILI 9
CAPSULES OR SLIME LAYERS 10
THE PERIPHERY OF THE BACTERIAL CELL 12
THE CELL WALL 12
BACTERIAL CELL MEMBRANE 14
L FORMS:-PROTOPLASTS AND SPHEROPLASTS 15
THE INSIDE OF THE CELL 17
CYTOPLASM 17
INCLUSION BODIES 17
NUCLEAR MATERIAL 17
PLASMIDS 19
RIBOSOMES 19
ENDOSPORES 19

SELF ASSESSMENT 22

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OBJECTIVES

The objectives indicate what you should know, understand and be prepared to explain upon completion
of this module. The self-assessment question will enable you to judge your understanding of the
maternal.

Upon completion of this module, the student should be able to:

1. given a diagram of a bacterium, indicate the following parts of the bacterial cell: flagellum, fimbria
(pilus), capsule, cell wall, cell membrane, cytoplasm, nuclear material, plasmids, ribosomes and, if
applicable, metachromatic granules and spores.

2. indicate which of the parts mentioned in 1. are essential for bacterial cell survival.

3. compare and contrast prokaryotic and eukaryotic cell structures.

4. describe briefly flagellar structure and function.

5. state the differences between true motility and Brownian motion.

6. define: peritrichous, monotrichous.

7. state how flagella and fimbriae may be seen.

8. .give the functions of fimbriae (pili) .

9. give the advantage to a cell of a capsule.

10. state how a bacterial cell capsule can be detected.

11. give two functions of the bacterial cell wall.

12. state the difference between the gram-positive and gram-negative cell walls.

13. state the relationship of the structure of the bacterial cell wall to the gram stain reaction.

14. state two functions of the bacterial cell membrane.

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15. state the difference between protoplasts and spheroplasts.

16. edscribe the structure of mesosomes.

17. . describe the character of the bacterial cytoplasm.

18. state the type of organelles present in the cytoplasm.

19. give the chemical composition of metachromatic granules.

20. describe how metachromatic granules are detected w1thin the cell.

21. give another name for metachromaic granules.

22. state four major d1fferences between the eukaryotic and prokaryotic nucleus.

23. name the process which describes bacterial division.

24. give the basic structure of the nuclear _region, i.e. double-stranded DNA, circular.

25. describe plasmid structure and significance.

26. state the difference between the prokaryotic and eukaryotic r1bosome.

27. state the usefulness of r1bosomes regarding control of bacterial disease.

28. state the general function of spores.

29. name the genera having endospores.

30. define endospore and free spore.

If you have studied this subject previously, you may test your ability us1ng the self assessment questions. If
you are able to obtain 90% or greater, you may choose not to do the unit and merely review the sections, or
parts of sections, where weakness may exist. If you obtain less than 90%, 1t is recommended that the module
be done in its entirety, stressing areas where more review is needed.

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RESOURCE MATERIAL

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MICROBIOLOGY
Ol4-l-MI
INTRODUCTION TO THE BACTERIAL CELL

The bacterial cell is profoundly different from the eukaryotic cell. This dissimilarity allows us to harm only the
bacterial cell by targeting with antibiotics, structures unique to bacteria. As well, some cell features make it
easy to separate bacteria into groups based upon differential staining properties. Still other distinctive cell
structures aid in its identification.

For even Just these few reasons, an examination of the bacterial cell structure is useful. However, the bacterial
cell is also worth studying because it can carry out all the functions of higher organisms using its "primitive"
cell organelles.

OVERVIEW OF THE BACTERIAL CELL

The bacterial cell will be examined first from its outside looking at flagellae, fimbriae and capsules; through
its "shell" consisting of the cell wall and membrane; and finally into its interior where the cytoplasm, nuclear
material, ribosomes, and other organelles may be found. Figure 1 depicts a stylized representation of these
structures.

Of all these structures, only the cell membrane, the nuclear region and ribosomes are essential for cell
survival. The others may be absent depending upon a given organism's needs.

Most information about bacterial structure comes not from light microscopy but from electron microscopy.
Figure 2 shows some of the detail that can be seen ~n an electron micrograph.

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THE OUTSIDE OF THE CELL

FLAGELLA (sing. flagellum)

Flagella are found on some but not all bacteria. They are delicate hair-like structures which have a simpler
chemical composition than the complex structure of eukaryotic flagella and cilia.

These long simple proteinaceous filaments originate in the interior of the cell and extend through the cell wall.

Using a rotary motion, flagella are able to propel the bacterium from one place to another. True bacterial cell
motitity is directional (cell moves from one point to another). Whereas brownian motion is the random
vibration of microscopic particles (including non-motile bacteria) by the energy from water molecules.

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Features used for identification include flagellar:

o presence or absence
o number
o distribution around a cell.

Because there appendage

are so fine, about 0.2 µm wide, they cannot be resolved by the light microscope. Certain special stains,
however, cot the flagellar surface, making the filament physically larger and therefore visible. Flagella are also
visible using the electron microscope.

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FIMBIAE AND PILI ( sing fimbria pilus)

Fimbriae are short filamentous appendages, straighter and more numerous then flagella. They are found on
some, but not all, bacterial cells. Unlike flagella are not involved in locomotion.

Fimbriae arise in the cytoplasmic membrane and penetrate through the cell wall to the exterior where several
hundred are distributed around the cell periphery. An electron microscope must be used to see fimbriae
Functlon

1. Fimbriae are used by the cell to adhere to host cell surfaces. This adherence is considered a virulence
factor since it encourages colonization by pathogens. For example, the organism causing gonorrhea,
Neisserzia gonorrhoeae, is thought to be virulent because it can adhere to the genital tract epithelial
cells

2. Fimbriae are thought to link together forming a layer of bacteria on fluid surfaces thus increasing
oxygen absorption at the air-fluid interface This feature is advantageous for growing bacteria which
require a lot of oxygen

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Some fimbriae are different and are designated as sex pill.. These special fimbriae elongate to form a hollow
conjugation tube stretching from one bacterium to another. This conjugation tube transports genetic
information from a donor to recipient cell.

CAPSULES OR SLIME LAYERS

Some, but again not all, bacteria secrete a slimy gelatinous substance which surrounds the cell outside the cell
wall. This substance is usually composed of polysaccharide or (polysaccharide-protein).

A slime layer is an unorganized accumulation of the polysaccharide on the cell surface capsules are more
organized and better defined accumulations of material.

Bacteria having capsules (Figure 9A) are more virulent than those without (Figure 9B) because phagocytes are
unable to catch and ingest encapsulated cells. Capsules also protect the cell against bactericidal substance in
the blood.

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Laboratory Significance

Organisms with capsular material produce colonies which have a very mucoid appearance. When touched with
an inoculating wire, they are stringy in texture. This feature is often an identification aid.

Capsules stain poorly but, if the encapsulated organism is suspended in an India Ink solution, the capsule will
displace the ink particles producing an empty space around the organism. The capsule is then said to be
negatively stained since the capsule now appears as a retractile halo around the cell.

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THE PERIPHERY OF THE BACTERIAL CELL

THE CELL WALL

Chemically, the bacterial cell wall is unique to prokaryotes. This special chemical is peptidoglycan (also called
murein or mucopeptide). It forms a giant macro –molecule which surrounds the entire cell in a rigid mesh,
providing shape and strength

The “glycan” part of the molecule ae the two amino sugars, NAG (n-acetyl glucosamine) and NAM (n-acetyl
muramic acid) which are alternated in chains. These chains are then cross-linked by the peptide portion
-strands of amino acids.

The diagram is intended only to give an idea of the complexity of the structure.

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As mentioned, the cell wall peptidoglycan is responsible for cell shape and strength.

The three basic bacterial cell shapes are:

o cocci (sing. coccus)
o bacilli (sing. bacillus) or rods
o spiral or he1ical shaped

Because most bacteria have a higher 1nternal pressure than the surrounding environment, a strong cell wall is
essential; otherwise, the cell would take up too much water by osmosis and burst. An analogy can be made
between a bacterial cell without its wall and a bicycle tire with only an inner tube.

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Laboratory Significance

Variations in cell wall structure allows us to differentiate most bacteria 1nto gram-positive and negative cells
using the gram stain.

The gram-positive cell wall is very thick, uniform and composed of up to 90% peptidoglycan.

In contrast, the gram-negative wall is thinner but considerably more complex. Its thin layer of peptidoglycan
comprises less than 20% of the wall and exterior to the peptidoglycan layer is an outer wavy and wrinkled
layer of lipopolysaccharide (LPS).

BACTERIAL CELL MEMBRANE

The bacterial cell membrane, a fragile structure hav1ng no mechanical strength, is located Just inside the rigid
cell wall. Its structure is very similar to that of a eukaryocytic cell having a lipid bilayer with the interspersed
proteins.

Bacterial membrane do lack sterols such which stabilize eukaryotic membrane; interestingly, sterols are found
in the membrane of the one bacterial genus-Mycoplasmas, organisms wh1ch survive and multiply w1thout a
cell wall.

As with higher life forms, the bacterial cell membrane controls the transport of most compounds entering or
leaving the cell. The bacterial cell membrane also contains enzymes needed for bacterial respiration; in
many ways the bacterial membrane is analogous to the mitochondrion in eukaryotes.

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Mesosomes

Mesosomes are relatively large, irregular invaginations of the cell membrane which project into the interior of
the cell of some, but not all bacter1a.

Some investigators be1ieve the mesosome increases the membrane surface area, functioning as an area for cell
enzyme activity and as a site for DNA attachment. However, others have indicated that mesosomes are
artifacts and have no function.

L FORMS: PROTOPLASTS AND SPHEROPLASTS

Some bacter1a exist naturally without cell walls these are the previously mentioned Mycoplasmas. Ordinary
cell-wall bacteria may be converted to living cells without cell walls called L-forms. These cell-defective form
may be divided into two groups, protoplasts and spheroplast.

Protoplasts:

Protoplasts are intact gram-positive bacaria minus a cell wall. How these cells lose their walls and yet remain
viable is useful to understand.

Agents such as lysozyme are able to hydrolyse the amino acid link between NAM and NAG in the
peptidoglycan. As a result the cell wall is stripped away leaving the protoplasm surrounded only by the thin
fragile cytoplasmic membrane. Because the membrane has little mechanical strength and because of the
internal pressure within the cell (greater solute concentration within the cell than w1thout*), the cell takes in
too much water through its semi-permeable membrane and literally bursts. Only the empty ghosts of
membranes remain.

* Cell is hypertonic to environment

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However, if the gram-positive cell is placed .in a solution having an osmotic pressure equivalent to that in the
cell's interior, lysis of the cell does not occur and the cell survives. A protoplast is thus created by action of
lysozyme in a solution isotonic to the cell interior.

Penicillin also is
able to induce protoplast formation because it also acts on the cell wall.

Spheroplasts

Because gram-negative bacteria have an outer lipopolysac charide membrane layer, lysozyme causes less
damage to the peptidoglycan. Spheroplasts, therefore are gram-negative bacteria with remnants of cell wall
material adhering to the cell. As a result they are less osmotically fragile than protoplasts .

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THE INSIDE OF THE CELL

CYTOPLASM

The cytoplasm is that watery fluid in the interior of the cell containing nucleic acids, proteins, carbohydrates,
1ipids, inorganic ions, ribosomes, inclusion bodies, nuclear materials and endospores

Unlike the eukaryotes, the cytoplasm of prokaryotes is less complex and has no endoplasmic reticulum or
mitochrondria.

INCLUSION BODIES

There are numerous inclusions within the cell such as lipid droplets, polysaccharides, glycogen, starch
deposits and inorganic substances.

Metachromatic Granules

These granules are inorganic phosphates which are used for food storage depot and a source of energy.
Metachromatic granules, also called volutin, are characteristically found in species of the genus
Corynebacteria The major pathogen in this genus is Corynebactenum diphtheriae, the causative agent of
diphtheria (note spelling)

A special staining procedure, Albert's stain, which colours the vegetative cell green and the granules black is
used to detect the granules.

Polychrome (or aged) methylene blue is also for staining these granules. This dye stains differentially by using
the principle of metachromasia, which means that the dye itself may change colour. In this case, the vegetative
cell appears blue but granules turn pink.

NUCLEAR MATERIAL

The nuclear region of prokaryotes is distinctly different from that of eukaryotes. For example, the prokaryotic
nuclear material

o lacks a nuclear membrane

o lacks mitotic apparatus
o lacks a nucleoius
o has a single chromosome

The single chromosome is a closed loop or circle of double-stranded DNA and is normally tightly called
within the cell during replication; this material uncoils forming a one micrometer loop of DNA

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Bacterial cells divide by the simple process of binary fission into two identical daughter cells each containing
a copy of the chromosome.

Simply stated, the process is as follows (F1gure 17):

The nuclear material of the parent cell doubles and the cell elongates.

The nuclear material divides. Ingrowth of plasma membrane and cell wall occurs.

A septum divides the protoplasm into two equal parts.

The cell wall septum splis the cell into two daughter cells.

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PLASMIDS

Plasmids are small extrachromosomal bits of circular DNA which replicate independently of the nuclear
region. They do not carry information essential for the life of the bacterium and can be gained or lost without
harm to the organism. Plasmids are important since they convey genetic information, most particularly
resistance to various drugs and antibiotics.

RIBOSOMES

Bacterial cell ribosomes have a similar structure to the eukaryotic cell except that the bacterial ribosome has a
lighter sedimentation constant of 70 S units (Svedberg Units) as compared to the 80 S units of the eukaryote.

In electron micrographs of the bacterial cell, ribosomes are seen as thousands of small granules scattered
throughout the cytoplasm. This dense packing of ribosomes in the cytoplasm indicates a high rate of
metabo1ic activity and protein synthesis.

Since bacterial ribosomes are different from those of eukaryotes, drugs may be directed specifically against
the activity of the bacterial ribosome leaving human ribosomes unaffected. For example, antibiotics such as
tetracyc1ine inhibit bacterial growth by ribosome damage but have little toxicity for the host.

ENDOSPORES

Endospores are refractile, round to oval bodies formed within the bacterial vegetative (ordinary living cell
structure Without a spore) cell. The spore structure is complex. A central core contains the nuclear material,
cytoplasmic membrane and all other essential life structures. The core is surrounded by a cortex which is in
turn surrounded by an outer coat.

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Depending upon the bacterial species, spores mayor may not bulge outside of the cell. Eventually the
vegetative cell disintegrates leaving free or mature spores. Endospore formation is i11ustrated 1n F1gure 20.
The process is as follows:

The nuclear material doubles and divides. The condensed nucleoid moves toward one end of the cell.

The plasma membrane invaginates to form the spore septum

The septum grows toward the tip of the cell unti1 the entire forespore is surrounded.

The spore coats are formed producing an endospore.

The mother cell lyses releasing a free spore.

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The endospore is the most resistant life form known. Spores permit the cell to survive adverse environmental
conditions such as heat, chemicals, drying, x-rays, ultraviolet rays, etc. Once conditions are again favourable
for growth, the spore will germinate into a vegetative form. Spores are not a method of reproduction since
each spore produces one vegetative cell.

Laboratory Significance

Many sterilants and disinfectants are evaluated on their ability to destroy or inactivate spores. Spores are
found in two major bacterial genera: the Bacillus species* and the Clostridium species. Both genera are gram-
positive rods (bacilli). The shape and position of spores within the cells are useful for identification.

Because the spore coat is so thick, special stains must be driven into the spore, usually by heat, so the spore
may be seen. When gram staining is used, spore-bearing bacteria will show only an unstained area whrre the
spore is located.

*N.B.: The genus name is spelled with a capital B. The word bacillus refers to a rod-shaped bacterium.

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 MICROBIOLOGY
014-l-MI
INTRODUCTION TO THE BACTERIAL CELL

SELF ASSESSMENT

MARKS

[3] 1. Which of the following structures are considered essential for bacterial cellsurvival:

cell wall, cell membrane, flagella, fimbriae, capsule, ribosomes, nuclear material, plasmids?
[2] 2. Describe Brownian motion and explain why is it not true motility.

[2] 3. Why does the bacterial cell need a rigid cell wall?

[ 4] 4. Match the structures in Column A with the statements in Column B:

COLUMN A COLUMN B
i) Spheroplasts A) Cell wall completely absent
ii) Protoplasts B) Remnants of cell wall
C) Gram positives
D) Gram negatives
E) Lysozyme induced
F) L-form

[1]5. What chemical, unique to prokaroytes, is found in the bacterial cell wall?

[1]6. What is the effect of lysozyme on the bacterial cell?

[1]7. What is an L-form bacterium?

[1]8. What is the chemical composition of metachromatic granules?

[1] 9. gi.ve another name for metachromatic granules.

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MARKS

[1] 10. How are metachromatic granules detected within the cell?

[1] 11. What are plasmids?

[1] 12. How do the ribosomes of bacteria and yeasts differ?

[1] 13. What structures do Bacillus and Clostridium species have that is unique?

[4] 14. Match the bacterial cell functions listed in Column A with the appropriate parts of the cell in Column
B. Some parts may be used more than once. (1/2 mark each).

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QUESTION 15 -20
[3] This question consists of four lettered headings.

Following these is a list of words or phrases with the letters A, B, C, D beside each.

For each numbered word or phrase BLACKEN the appropnate lettered space on the computer sheet using the
following key:

[4] 21. Match the parts of the bacterial cell listed in Column A with the statements in Column B. (1/2 mark
each)

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MARKS:
[20] 22. Fill in the following chart comparing eukaryotes and prokaryotes. (1 mark for each comparison)

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MARKS

[9] 23. Label the following diagram if the bacterial cell using the items listed on this sheet. (1 mark for each
correct answer)

6O TOTAL

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MICROBIOLOGY
014-1-MI
INTRODUCTION TO THE BACTERIAL CELL

SELF ASSESSMENT ANSWERS

1. Cell wall -not essential, for example: mycoplasma and L-forms survive. Cell membrane -essential
Flagella -not essent1al, many bacteria do not have them fimbriae (pi1i) -not essential many bacteria do not
have them Capsule -not essential Ribosomes -essential, needed for protein synthesis Nuclear material
-essential Plasmids -not essential

2. Brownian motion -It is not true moti1ity since it is not directional. It is random motion of microscopic
particles generated by the energy in water molecules.

3. Provides shape Provides osmotic protect1on

4.

5. Peptidoglycan (murein)

6. Causes the cell to burst

7. A form of bacterium deficient in all or part of its cell wall.

8. Inorganic phosphates

9. Volutin

10. By the use of a granule stain such as Albert's stain or polychrome methylene blue.

11. Plasmids are extra-chromosomal bits of DNA. Plasmids transfer genes for resistance to antibiotics,.

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12. Bacteria have 70 S r ibosomes whereas yeasts (eukaryotes) have 80 S ribosomes.

13. Both genera have spores.

14.

2l.

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[20] 22. Fill ion the following chart comparing eukaryotes and prokaryotes. (1 mark for each comparison).

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23. ANATOMY OF THE PROKARYOTIC CELLS – ANSWERS.

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