Procaryotic and Eucaryotic

Microorganisms
and
Bacterial Diversity
215205
Jomkhwan Meerak, Ph.D

Procaryotic
Microorganisms & Diversity

1
 Sizes of Microbes
• Virus - 10 →1000 nanometers *
• Bacteria - 0.1 → 5 micrometers **

(Human eye ) can see .1 mm (1 x 10 -3 m)

How can we see Microbes???

Eucaryotes Procaryotes Noncellular

:Bacteria :Viruses
:Animal cells :Protozoa :Archaea
:Plant cells :Unicellular algae
:Multicellular :Some fungi (yeasts &molds)
algae
microorganisms

2
 Prokaryotic Cells: An Overview
• Prokaryotes
– Do not have membrane surrounding their
DNA; lack a nucleus
– Lack various internal structures bound with
phospholipid membranes
– Are small, ~1.0 µm in
diameter
– Have a simple structure
– Composed of bacteria
and archaea

ความสั มพันธ์ procaryote กับ eucaryote

เชื" อว่ า organelles บางอย่ างของ eucaryote วิวฒ
ั นาการมาจาก
procaryotic cell ทีถ" ูก endocytosis เข้ าไปใน eucaryotic cell
Endosymbiotic cyanobacteria-->chloroplast

3
 **Origin of Mitochondria **
มาจาก free living aerobic bacteria
------> mitochondria

Prokaryotic vs. Eukaryotic

Http://micro.magnet.fsu.edu/cells.html

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 Prokaryotes Eukaryote Eukaryote
Characteristic Animal Plant
1. nuclear membrane no yes yes
2. plasma membrane yes yes yes
3. cell wall yes no yes
4. ribosomes yes yes yes
5. Endoplasmic reticulum no yes yes
6. Golgi complex no yes yes
7. lysosomes no yes yes
8. peroxisomes no yes yes
9.nucleolus no yes yes
10.mitochondria no yes yes
11.chloroplasts no no yes
12.microtubules no yes yes
13.actin filaments no yes yes
14.chromosome single multiple multiple

The Prokaryotic Cell: Shape

• Typically fixed shape due to cell wall

– Peptidoglycan (murein) in bacteria
– Pseudomurein in archaea
• Rod: bacillus (E.coli)
• Coccus: round, spherical
– Diplococci (N. meningitidis)
– Streptococci (S. pyogenes)
– Tetrades (Micrococci)
– Sarcinae
– Staphylococci (Staphylococcus epidermidis)
• Spiral
– Fixed: spirilla
– Flexible: spirochetes (Treponema pallidum)
10

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Rods Cocci

11

Spiral Shaped

12

6
 Other Prokaryotic Shapes
• Unusual
– Star-shaped
– Squares
– Triangular
• Pleomorphic
– Within a population
various shapes
(Corynebacteria)
• No fixed shape
Shape is influenced by
– Cellwall-less: environmental conditions, age of
Mycoplasma/Ureaplasma culture, antibiotic pretreatment!
13

SIZE OF BACTERIA
• Unit for measurement :
Micron or micrometer,μm:
1μm=10-3mm
• Size:
Varies with kinds of bacteria,
and also related to their age and
external environment.

 Cocci: sphere, 1μm

 Bacilli: rods , 0.5-1 μm in width -3 μm in length
 Spiral bacteria: 1~3 μm in length and 0.3-0.6 μm in width

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 การสื บพันธุ์ของ procaryotes
• Asexual reproduction
พวกทีเป็ นเซลล์เดียว สื บพันธุ์แบบ
Binary fission

พวกทีเป็ นเส้นสาย สื บพันธุ์แบบ

Fragmentation

PROKARYOTIC CELL STRUCTURE

Keywords:
- prokaryotic cell
- archaebacteria vs. eubacteria
- cell wall, plasma membrane
- nucleoid
- flagella, pili
- gram positive bact., gram negative bact.
-inclusion body
-endospore

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 Prokaryotic Cells Structure

[INSERT FIGURE 3.2]

General component of Prokaryotic cell

- Capsule : Found in some bacterial cells, this additional outer
covering protects the cell when it is engulfed by other
organisms, assists in retaining moisture, and helps the cell
adhere to surfaces and nutrients.
- Cell Wall : Outer covering of most cells that protects the
bacterial cell and gives it shape.
- Cytoplasm : A gel-like substance composed mainly of water
that also contains enzymes, salts, cell components, and
various organic molecules.
- Cell Membrane or Plasma Membrane : Surrounds the cell's
cytoplasm and regulates the flow of substances in and out of
the cell.

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General component of Prokaryotic cell
- Pili : Hair-like structures on the surface of the cell that
attach to other bacterial cells. Shorter pili called fimbriae
help bacteria attach to surfaces.
- Flagella : Long, whip-like protrusion that aids in cellular
locomotion.
- Ribosomes : Cell structures responsible for protein
production.
- Nucleiod Region : Area of the cytoplasm that contains the
single bacterial DNA molecule.
- Plasmids : Gene carrying, circular DNA structures that are
not involved in reproduction.

External Structures of Prokaryotic Cells

• Glycocalyces
• Flagella
• Fimbriae and pili

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 Structures External to Cell wall:
Glycocalyces
Outer most layer
Outside cell wall  Extracellular
Usually sticky polysaccharide allows cell
Composed of mostly to attach
of carbohydrates,  Capsules prevent
sometimes of protein phagocytosis
 Capsule: neatly
organized
 Slime layer: unorganized
and loose
 Biofilm or extracellular
polymeric substance
(EPS) 21

External Structures of Prokaryotic Cells

• Glycocalyces
– Gelatinous, sticky substance surrounding the
outside of the cell
– Composed of polysaccharides, polypeptides,
or both

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 Two Types of Glycocalyces
– Slime layer
• Loosely attached to cell surface
• Water soluble
• Protects cells from drying out
• Sticky layer that allows prokaryotes to
attach to surfaces

– Capsule
• Composed of organized repeating units
of organic chemicals
• Firmly attached to cell surface
• Protects cells from drying out
• May prevent bacteria from being recognized
and destroyed by host

[INSERT FIGURE 3.5]

12
Capsule staining:
negative stain technique --> Indian ink or nigrosin

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 Flagella
• Are responsible for movement
• Have long structures that extend beyond cell
surface
• Not all prokaryotes have flagella
• Composed of filament, hook, and basal body
• Flagellin protein (filament) is deposited in a helix
at the lengthening tip
• Base of filament inserts into hook
• Basal body anchors filament and hook to cell wall
by a rod and a series of either two or four rings of
integral proteins
• Filament capable of rotating 360º

Bacterial Flagella Structure

Figure 3.5a

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 Bacterial Flagella Structure

Figure 3.5b

Bacterial flagella

A-Monotrichous

B-Lophotrichous

C-Amphitrichous

D-Peritrichous

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Arrangements of Bacterial Flagella

Arrangements of Bacterial Flagella

Figure 3.6b

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Arrangements of Bacterial Flagella

Figure 3.6c

Flagella examination

• Proteus vulgaris ( growing at 27-30 ºC)

• 1. Staining
• 2. Hanging drop technique
• 3. Culture / stab/ semi solid media

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1. Stain

Hanging drop technique

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Klebsiella Proteus

stab

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Function of Bacterial Flagella
• Rotation propels bacterium through environment
• Rotation can be clockwise or counterclockwise;
reversible

Bacterial Movement

• Bacteria move in response to stimuli (taxis)

– Runs – movements of cell in single direction for
some time due to counterclockwise flagellar
rotation; increase with favorable stimuli (positive
chemotaxis, positive phototaxis)
– Tumbles – abrupt, random, changes in direction
due to clockwise flagellar rotation; increase with
unfavorable stimuli (negative chemotaxis,
negative phototaxis)

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 **Taxis **
• Chemotaxis
• Phototaxis
• aerotaxis
• Geotaxis
• magnetotaxis

Other Bacterial Projections

• Axial Filaments—
bundles of filaments
that arise at the end
of the cell beneath the
outer sheath. Spiral
around the entire cell

• Bacteria with axial

filaments move in a
corkscrew shape
motion

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 Axial filament
• Spirochetes มี endoflagella หรื อ
เรียกว่ า
Axial filament
• Treponema pallidum สาเหตุ
syphilis
• Treponema บางชนิด commensal
ในช่ องปากของคน

Motile by gliding
• ไม่มี flagella แต่เคลือนทีได้บน
solid surface
• ตัวอย่าง filamentous cyanobacetria,
myxobacteria

22
 Fimbriae and Pili
• Nonmotile extensions
• Fimbriae
– Sticky, proteinaceous, bristlelike projections
– Used by bacteria to adhere to one another, to
hosts, and to substances in environment
– May be hundreds per cell and are shorter
than flagella
– Serve an important function in biofilms

Fimbriae Versus Flagella

Figure 3.9

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 Pili
• Long hollow tubules composed of pilin
• Longer than fimbriae but shorter than
flagella
• Bacteria typically only have one or two per
cell
• Join two bacterial cells and mediate the
transfer of DNA from one cell to another
(conjugation)
• Also known as conjugation pili or sex pili

Pilus Versus Fimbriae

Figure 3.10

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 Bacterial Cell Surface

Structure of Bacteria’s Cell Walls

Prokaryotic Cell Walls

• Provide structure and shape and protect cell
from osmotic forces
• Assist some cells in attaching to other cells
or in eluding antimicrobial drugs
• Not present in animal cells, so can target
cell wall of bacteria with antibiotics
• Bacteria and archaea have different cell wall
chemistry

25
• Bacterial Cell Walls

– Most have cell wall composed of peptidoglycan

– Peptidoglycan is composed of sugars, NAG,
and NAM
– Chains of NAG and NAM attached to other
chains by tetrapeptide crossbridges
• Bridges may be covalently bonded to one another
• Bridges may be held together by short connecting
chains of amino acids
– Scientists describe two basic types of bacterial
cell walls: Gram-positive and Gram-negative

External Structures of Prokaryotic Cells

[INSERT FIGURE 3.12]

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 External Structures of Prokaryotic Cells

[INSERT FIGURE 3.13]

• Bacterial Cell Walls

– Gram-positive cell walls

• Relatively thick layer of peptidoglycan
• Contain unique polyalcohols called teichoic acids
– Some covalently linked to lipids, forming lipoteichoic
acids that anchor peptidoglycan to cell membrane

• Retain crystal violet dye in Gram staining

procedure; so appear purple
• Up to 60% mycolic acid in acid-fast bacteria helps
cells survive desiccation

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• Bacterial Cell Walls

– Gram-negative cell walls

• Have only a thin layer of peptidoglycan
• Bilayer membrane outside the peptidoglycan
contains phospholipids, proteins, and
lipopolysaccharide (LPS)
• May be impediment to the treatment of disease
• Appear pink following Gram staining procedure

Gram Positive vs. Gram Negative

Plasma
Membrane

LPS is another important component of G (-)

LPS stimulates the Immune System

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 Hans Christian Joachim Gram, 1852-1938

The test itself is useful in classifying two

distinct types of bacteria based on the
structural differences of their cell walls.

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**KOH test for Gram negative and Gram
positive bacteria**
“String test”

3% KOH - bacteria -- 1 นาที

Gram negative …> สายหนืดๆ
ของ DNA

Damage to the Cell Wall

NAG NAM NAG

Penicillin

Lysozyme

NAG NAM NAG

• Penicillin: inhibits transpeptidases

• Lysozyme: breaks glycosidic bond between NAM and
NAG
• Autolysins: self destruction of peptidoglycan, important for
cell divisions

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 Atypical Cell walls
Acid Fast Cell walls
• Rich in mycolic acids
• E.g. Mycobacterium spec
• Hard to penetrate
• Require heat for staining
Archaea
• No peptidoglycan (murein) but
with pseudomurein
• Contains variety of specialized
polysaccharides and proteins
– Gram-positive archaea stain
purple
– Gram-negative archaea stain
pink

61

Gram variable
: Gram + ve เซลล์ อายุมากๆ อาจเปลีย" นเป็ น
Gram -ve เช่ น Bacillus
: หรื อจาก Gram - ve เปลีย" นไปเป็ น> Gram +ve
เช่ น Streptobacillus moniliformis

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แบคทีเรี ยทีไม่มี cell wall >> Mycoplasma
ใน cell membrane มี sterol ทําให้เซลล์มีความแข็งแรง

Talaro,K.,and Talaro, A.1993.

**Cell with no walls**

:** เลีUยง Gram + ใน isotonic solution
---> lysozyme แยก peptidoglycan (PG)
(break… -1,4 glycosidic bonds
ß

-----> “protoplast” เหลือ cell membrane

: **ถ้าแยก PG จาก Gram - จะได้>--->
“sphaeroplast”เหลือ cell membrane
+ outer membrane

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 (Lister Institute of Preventive Medicine)
: เลีUยงใน antibiotics, temperature shock,
osmotic shock -->inhibit cell wall synthesis เช่น
: Bacillus, Proteus, Streptococcus & Vibrio
: เซลล์เปลียนเป็ น ทรงกลมไม่มี cell wall
*Streptobacillus moniliformis เป็ น pleomorphic
Gram negative ....spotanously produces L-form

Periplasmic space or periplasm is a space

between the inner cytoplasmic membrane and external outer
membrane of Gram-negative bacteria

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 Prokaryotic Cytoplasmic
Membranes
• Structure
– Referred to as phospholipid bilayer; composed
of lipids and associated proteins
– Approximately half composed of proteins that
act as recognition proteins, enzymes,
receptors, carriers, or channels
• Integral proteins
• Peripheral proteins
• Glycoproteins
– Fluid mosaic model describes current
understanding of membrane structure

Prokaryotic Cytoplasmic Membranes

[INSERT FIGURE 3.15]

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Prokaryotic Cytoplasmic Membranes

• Function
– Energy storage
– Harvest light energy in photosynthetic
prokaryotes
– Selectively permeable
– Naturally impermeable to most substances
– Proteins allow substances to cross membrane
• Occurs by passive or active processes
– Maintain concentration and electrical gradient
• Chemicals concentrated on one side of the membrane or
the other
• Voltage exists across the membrane

หน้ า ทีของ cell membrane ของแบคทีเรีย

1. ควบคุม การผ่า นของสาร มีก ลไกทีอาจ

เป็ น
1.1 Simple diffusion
1.2 Diffusionโดยมี carrier protein
1.3 Active absorption มี permeases & ATP
ปั ม$ สารเข้า

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Prokaryotic Cytoplasmic Membranes

[INSERT FIGURE 3.17]

Prokaryotic Cytoplasmic Membranes

[INSERT FIGURE 3.19]

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2. เป็ นบริ เวณทีมีการ respiration ของเซลล์
:-electron transport system to generate
ATP

3. เป็ นทีติดของ flagella

4. Mesosome
- เป็ นบริ เวณทีมีการสร้าง septum
- เป็ นทีติดของ chromosome
เมือเกิด DNA replication

5. เกีย" วข้ องกับ Nitrogen fixation

เช่น Azotobacter
การตรึ ง Nitrogen พบเฉพาะ
ใน procaryote เท่านัUน

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 6. Internal membrane system

• Photosynthetic bacteria & Cyanobacteria

มี chromatophore เปลีย2 นแปลงมาจาก cell
membrane มีรงควัตถุ
ทําหน้าที2 สังเคราะห์แสง

Chromatophore

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 Cytoplasm of Prokaryotes
• Cytosol – liquid portion of cytoplasm
• Inclusions – may include reserve deposits of
chemicals
• Ribosomes – sites of protein synthesis
• Cytoskeleton – plays a role in forming the
cell’s basic shape
• Some bacterial cells produce dormant form
called endospore

Internal components

Nucleoid- with single, circular, supercoiled

DNA molecule

Many bacteria have plasmids

small, extrachromosomal, circular
piece of DNA

genes present are usually not required

but may be advantageous
(antibiotic resistance, resistance to metals)
Now used for genetic engineering

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Bacterial DNA:  Chromosomal DNA
 Plasmid

Prokaryotic Nucleoid

• Bacterial chromosome
• Contains the essential
genetic information
• Circular double-
stranded DNA
Stabilized by histone-
like proteins (not by
histones)
• No nuclear envelope!! 80

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 Prokaryotic Plasmids
• Small circular double-stranded
DNA that can multiply
independently
• Not essential under normal
physiological conditions
• Contain additional genes often
involved in pathogenesis
– virulence factors
– antibiotic resistance
– toxic metal resistance
• Copy number varies (a few –
hundreds)
– Can be exploited for recombinant
protein production

81

Classification of plasmids is by function

5 main classes:
Fertility-F-plasmids, which contain tra-genes. They are capable of
conjugation (transfer of genetic material between bacteria which are
touching).

Resistance-(R) plasmids, which contain genes that can build a resistance

against antibiotics or poisons and help bacteria produce pili. Historically
known as R-factors, before the nature of plasmids was understood.

Col-plasmids, which contain genes that code for (determine the production
of) bacteriocins, proteins that can kill other bacteria.

Degradative plasmids, which enable the digestion of unusual substances,

e.g., toluene or salicylic acid.

 Virulence plasmids, which turn the bacterium into a pathogen (one that
causes disease).

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Commercial plasmid for genetic engineering

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 Prokaryotic Ribosomes
• 70S ribosomes (30S + 50S subunit)
– S = Svedberg unit (sedimentation rate upon centrifugation)
– smaller than eukaryotic ribosome
– sediment differently

• Consist of proteins and ribosomal RNA

• Site of protein synthesis
• Contain 16S rRNA on 30S subunit

Important for Classification and Identification!

85

Bacterial Endospores: Endurance

Forms
Spore

Sporulation Germination

Vegetative Form

• Terminal
• Subterminal
• Central 86

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 Bacterial Endospores: Unique Endurance forms

• Sporulation is a complex
process
• Triggered under
unfavorable conditions
• Very low water content
• Spore is multilayered
• Resistance through spore
coat (protein layer)
• Can survive thousands of
years
• Germination is triggered
under favorable conditions
87

Endospores  Identification
(spores) of Bacteria
 Pathogenesis
 Resistance

• Dormant cell
• Resistant to adverse • Produced when starved
conditions • Contain calcium dipicolinate
- high temperatures DPA, Dipicolinic acid
- organic solvents • Bacillus and Clostridium

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Endospore staining

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 Prokaryotic Inclusions and
granules
• Storage granules (lipids, carbohydrates,
phosphate etc)
• Caroxysomes (important for carbon
dioxide fixation in photosynthesis)
• Gas vesicles for bouyancy
• Magnetosomes

91

• Inclusion body มี membrane ล้อมรอบได้แก่

• 1. Poly-ß-hydroxybutyrate (PHB)
• 2. glycogen
• 3. gas vesicles
• 4. magnetosome

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 Inclusion bodies of PHB

Gas vesicles

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 Magnetosome
• Magnetotactic bacteria
• Fe2O 3 (iron oxide)
• Aquaspirillum magnetotacticum
• Magnetotaxis/เคลือนทีตอบสนองสนามแม่เหล็กโลก
magnetic field
• พบ magnetosome ใน bacillus-shaped structure
ในก้ อนอุกกาบาตจากดาวอังคาร

Granules ไม่มี membrane หุม้

• 1. Sulfur granules of
photosynthetic
bacteria
2. polyphosphate granules /
หรื อเรียก metachromatic
granules หรื อ volutin
granules

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 **Conidium**
• Filamentous bacteria เช่น
actinomycetes จีนสั Streptomyces
• มี chain of conidia
• ไม่ทนร้อน
• เป็ น reproductive structure

**Cysts**
• Gram negative bacteria บางชนิดมี cysts
• Resting cell ทนแล้ งไม่ ทนร้ อน ไม่ ทนสารเคมี
• ตัวอย่างแบคทีเรี ยทีมี cyst ได้แก่ Azotobacter
• Cyst ….germinate ได้ 1 cell
• ดังนัUนจึงไม่เป็ น reproductive structure
• : ไม่มี dipicolinic acid

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