Lecture Outline 

Biology 2230, Microbiology          

 

Chapter 1  The Microbial World and You

 

Naming and Classifying Microorganisms

1735 – Linnaeus – set up binomial nomenclature

Genus + specific epithet = species

Ex.  Escherichia coli

After first citation can use E. coli

 

Types of Microorganisms (Fig. 1.1)

 

A.  Bacteria

Unicellular

Prokaryotes (no nucleus)

Three shapes

1.  Bacillus à rods

2.  Coccus à spherical

3.  Spiral à corkscrew

Cell walls of peptidoglycan

Produce asexually by binary fission

 

B.  Archaea

Unicellular

Prokaryotes

Some have cell walls (but not of peptidoglycan)

Live in extreme environments

3 groups

1.  Methanogens – produce methane

2.  Extreme halophiles – in very salty environments

3.  Extreme thermophiles – in hot springs

 

C.  Fungi

Unicellular or multicellular

Eukaryotes ( nucleus)

NO photosynthesis

Cell walls of chitin

Reproduce asexually or sexually

Two forms

1.  Yeasts – unicellular

2.  Molds – multicellular, filamentous

 

D.  Protozoa

Unicellular

Eukaryotic

No cell walls

Reproduce sexually or asexually

Can be free living or parasitic


E.  Algae

Eukaryotes

Most unicellular

Photosynthetic

Sexual and asexual reproduction

Cell walls of cellulose

 

F.  Viruses

Very small

Acellular

All are parasitic

 

G.  Helminths

Flatworms and Roundworms

Parasitic

Multicellular

 

Classification of organisms

A.  Domain Bacteria

B.  Domain Archaea

C.  Domain Eukarya

1.  Kingdom Protista (Protozoa and Algae)

2.  Kingdom Fungi

3.  Kingdom Plantae

4.  Kingdom Animalia

 

History of Microbiology

 

A.  Early microscopy

 

1665 – Robert Hooke looks at cork with microscope and sees cells

 

1673 – Leeuwenhoek – sees live microbes (Fig 1.2)

 

B.  Spontaneous generation vs. biogenesis

Spontaneous generation – life can arise spontaneously from nonliving matter

Biogenesis – living cells can only arise from preexisting living cells

 

1668 – Redi – sealed jars of meat had no maggots

 

1745 – Needham – heated broths, put in covered flasks, grew microbes

 

1765 – Spallanzani – heated broth in flask after sealed à no growth

But others said no growth was due to lack of oxygen

 

1861 – Louis Pasteur

Swan neck flask experiment (Fig 1.3) à Biogenesis was correct

Also developed aseptic techniques (prevent contamination by

unwanted microorganisms) and pasteurization.


C.  The Germ theory of disease – microorganisms cause disease

 

1796 – Jenner – vaccine for smallpox

exposure to cowpox gives immunity to smallpox

 

1840’s – Semmelweiss

Had physicians wash hands

à drastically cut rates of puerperal fever

 

1860’s – Lister

Treated surgical wounds with phenol

à Drastically reduced incidence of infections

 

1876 – Robert Koch

Established Koch’s postulates

à Sequence of experimental steps for directly relating a specific microbe to a specific disease.

 

D.  Chemotherapy

 

1.  Synthetic drugs

1910 – Ehrlich

Developed salvarsan - “magic bullet” for syphilis

1930’s – sulfa drugs

 

2.  Antibiotics

1928 – Alexander Fleming 

Discovered penicillin (Fig. 1.5)

 

 

Beneficial activities of Microorganisms

 

A.  Recycling vital elements

Ex:  nitrogen fixation

 

B.  Sewage treatment

 

C.  Bioremediation

Break down toxins

Ex:  Exxon Valdez oil spill cleanup

 

D.  Insect pest control

Ex:  Bacillus thuringiensis kills agricultural pests

 

E.  Biotechnology

Producing foods and chemicals

 

F.  Genetic engineering

Gene therapy – Ex:  replace faulty cystic fibrosis gene,

Develop disease and drought resistance in crops


Relationship of microbes and humans

 

A.  Normal Microbiota (Fig 1.7)

Live on or in our bodies

Beneficial – prevent overgrowth of harmful microbes, produce vitamins

 

B.  Pathogens

Invade host, carry on life cycle, cause damage to the host

 

 

Emerging infectious diseases (caused by pathogens)

Bovine Spongiform encephalopathy (BSE)

Creutzfeldt-Jakob disease (CJD)

E. coli O157:H7

Invasive Group A Streptococcus “flesh-eating bacteria”

Ebola

Hantavirus

Acquired Immunodeficiency syndrome (AIDS)


Chapter 3  Observing Microorganisms through a Microscope 

 

Units of measurement (Table 3.1) Know metric units and metric equivalents

 

Microscopy:  The instruments (Fig 3.4) (Table 3.2)

 

A.  Brightfield Microscopy (Compound Light Microscopy)

See dark object on light background

Usually have to stain to increase contrast

 

B.  Darkfield Microscopy

Use opaque disc to create light object on a dark background

Gives better contrast

Use on unstained live specimens

 

C.  Phase-contrast microscopy

Better contrast

Use on unstained live specimens

Can see internal structures better

 

D.  Fluorescence Microscopy

Use fluorescent dyes

See glowing objects on dark background

 

E.  Electron Microscopy

Use electrons rather than light

1000X more magnification and resolution than light microscopes

 

1.  Transmission Electron Microscopy

Electrons transmitted though specimen

2-D view

 

2.  Scanning electron Microscopy

Electrons scattered off surface

3-D view

 

 

Relationship between sizes (Fig 3.2)

 

 

Preparation of specimens for light microscopy à staining

 

Stains = salt = Positive ion + negative ion

Only one ion is colored à chromophore

Non-colored ion à auxochrome

DRAW


Types of Staining

 

A.  Negative staining à chromophore is negative à uses acidic dyes

DRAW

 

 

 

 

 

 

 

 

B.  Positive staining à chromophore is positive à uses basic dyes

DRAW

 

 

 

 

 

 

 

 

 

Two types of positive staining

 

1.  Simple staining

Use single dye

Ex:  Crystal violet

 

2.  Differential staining

Use more than one dye

Used to distinguish different bacteria

Ex:  Gram stain (Crystal violet and Safranin) (Fig. 3.10)


Chapter 4  Functional Anatomy of Prokaryotic and Eukaryotic Cells

 

Overview – Prokaryotes vs. Eukaryotes

 

Prokaryotes

Eukaryotes

No nucleus

Nucleus

One circular chromosome

Multiple Chromosomes

No mem. enclosed organelles

Organelles

Cell wall of peptidoglycan

Cell wall, when present, is not peptido.

Divide by binary fission

Divide by mitosis

 

 

 

 

 

 

 

 

The prokaryotic cell

 

A.  Size

.2 – 2.0 mm diameter

2 – 8 mm length

 

B.  Shape

1.  Coccus - spherical

2.  Bacillus – rod shaped

3.  Coccobacilli - oval

4.  Spirals (Fig 4.4)

a.  Vibrios – comma-shaped

b.  Spirilla – corkscrew (rigid)

c.  Spirochetes – corkscrew (flexible)

5.  Pleomorphic – many shapes

 

C.  Arrangements

 

1.  Coccus (Fig. 4.1)

a.  Diplococci – pairs

b.  Streptococci – chains

c.  Tetrads – groups of 4

d.  Sarcinae – cube like groups of 8

e.  Staphylococci – grapelike clusters

 

2.  Bacilli (Fig 4.2)

a.  Diplobacilli – pairs

b.  Streptobacilli – chains

 

3.  Spirals – none


Structure of a typical prokaryotic cell (Fig. 4.6)

 

A.  Structures external to the cell wall

 

1.  Glycocalyx – gelatinous layer of polysaccharide and/or polypeptide

 

a.  Capsule

Organized

Firmly attached to cell

Protects cell from host immune cells

 

b.  Slime layer

Unorganized

Loosely attached

 

c.  Extracellular polysaccharide

Thin polysaccharide fibers

Use to attach to surfaces

 

2.  Flagella

 

a.  Arrangements (Fig. 4.6)

(1)  Monotrichous – single polar flagellum

(2)  Amphitrichous – tuft of flagella or single flagella at each end of cell

(3)  Lophotrichous – 2 or more flagella at one pole

(4)  Peritrichous – flagella all over cell

 

b.  Structure (Fig. 4.8)

Filament

Hook

Basal body

 

c.  Patterns of motility (Fig. 4.9)

Move by “runs and tumbles”

Runs–flagella rotates one way, run in straight line

Tumbles – flagella rotates opposite way, tumbles

Chemotaxis – movement toward or away from chemical

Phototaxis – movement toward or away from light

 

3.  Axial Filaments (Fig 4.10)

Flagella inside outer sheath

Corkscrew motion

Seen in spirochetes

 

4.  Fimbriae

Many short hairlike protein appendages

Function:  attachment


5.  Pili

Longer protein tube

Function:  transfer DNA from one cell to another

DRAW

 

 

 

 

 

B.  Cell Wall

           

1.  Function

a.  Protect cell from osmotic lysis

b.  Maintain shape of cell

c.  Anchorage for flagella

 

2.  Composition (Fig 4.13a)

Made of layers of peptidoglycan either alone or with other substances

Structure of peptidoglycan

DRAW

 

 

 

 

 

 

 

 

3.  Types of cell walls

 

a.  Gram-positive Cell Walls

DRAW

 

 

 

 

 

 

 

 

b.  Gram-Negative cell walls

DRAW


4.  Cell walls and the Gram stain mechanism

a.  Crystal Violet (primary stain) à stains all cells

b.  Iodine  (mordant àintensifies stain)

à forms large CV-I crystals

c.  Alcohol (decolorizer)

à seals off thick cell wall of Gram positive

à destroys outer mem & leaches stain on Gram neg.

d.  Safranin – (counterstain) à stains Gram negative pink

 

5.  Atypical cell walls

a.  Mycoplasma – no cell wall

b.  Archaea – no peptidoglycan, some no cell wall

 

C.  Structures internal to the cell wall

 

1.  Plasma membrane (Fig 4.14)

a.  Structure

Phospholipid bilayer

Proteins

Peripheral proteins

Integral proteins

Fluid mosaic model

DRAW

 

 

 

 

 

 

 

b.  Function

Selective permeability – certain molecules can pass through, others cannot

 

2.  Cytoplasm – substance inside the plasma membrane

 

a.  Nucleoid

Location of bacterial chromosome

 

b.  Plasmids

Small circular DNA molecules

Have 5 to 100 genes

Can carry genes for antibiotic resistance and toxin prod.

 

c.  Ribosomes (Fig. 4.19)

Sites of protein synthesis

Composed of protein and rRNA

70s (eukaryotic ribosomes are 80s)


d.  Inclusions – deposits in cell

 

(1)  Metachromatic (volutin) granules

Source of phosphate used to make ATP

 

(2)  Polysaccharide granules

Consist of glycogen or starch

Used for energy storage

 

(3)  Lipid inclusions

Usually poly-β-hydroxybutyric acid (PHB)

à unique to bacteria, used for energy storage

 

(4)  Magnetosomes (Fig. 4.20)

Iron oxide

Act like magnet

 

D.  Endospores

Dehydrated cells with thick wall

Not a reproductive process

Survival structure

à resistant to heat, drying, chemicals, radiation

à can survive boiling for 19 hours

Seen in the genera Bacillus and Clostridium

Ex:  Bacillus anthracis à causes anthrax

Clostridium tetani à causes tetanus

 

Sporulation – triggered by nutrient depletion (Fig. 4.21)

1.  Spore septum begins to form

2.  Plasma membrane surrounds DNA

3.  Spore septum surround isolated portion

4.  Peptidoglycan layer forms

5.  Spore coat forms

6.  Endospore freed from cell

 

Germination

Occurs when conditions turn favorable

Return to vegetative state

25 million year old endospores in amber have germinated