Prokaryotic Anatomy
2005 Summer Microbiology class

PROKARYOTIC ANATOMY


Syllabus and Laboratory Handouts
Summer Quarter
©David B. Fankhauser, Ph.D.,
Professor of Biology and Chemistry
             University of Cincinnati Clermont College,
Batavia OH 45103
Microbiology, Summer 2004
Microbiology Students,
Summer 2006
This page has been accessed Counter times since 15 July 2007. 
Microbiology Students,
Summer, 2004

Students performing a 
yeast plate count experiment
Prokaryotic Anatomy (by David Fankhauser)

PROKARYOTIC ANATOMY: CAPSULES, FLAGELLA, PILI

 

6/30/83, rvsd 8 July 1995, 25 June 99, 13 July 01, 7 July 03, 14 July 04, 11 July 05, 5 Apr 06, 9 July 07

TFC, 7th p 77-99, Alcamo, p. 87-, Atlas, pp 111-139, TFC, 8th p 76-96, Black 6th: 77-89, Bauman 2nd, 55-65

 

CELL SHAPE (2):            coccus, bacillus, spirillum, pleomorphic

ARRANGEMENT:     depends on how cells stick together after mitosis:

                                                    diplo, strepto-, tetrad, sarcinae, staphylo

                                                         

GLYCOCALYX (sugar + cup, flower) sticky, gelatinous layer, shown by negative stain. (Not in spirillum)

                          capsule           if firmly attached to bacterial cell (see p 61)

                          slime layer      so called if loose, unorganized

Extracellular Polymeric Substance = EPS, if composed of sugars: used for attachment especially

                          mucopolysaccharide:   polysaccharide [dextran] cross-linked with small peptides

 

Function:         buffers, protects fr phagocytosis, dehydration, adheres organism to substrate

                                       pathogenicity: B. anthracis + S. pneumoniae only pathogenic if encapsulated (smooth)

                                       adherence:       Klebsiella sticks to respiratory tract, S. mutans in forming dental caries.

 

“Ropy” milk, beer due to Alcaligenes viscolactis capsular material, ropy bread to B subtilis

 

Quellung reaction (swelling): swelling of capsule from specific antibody attack, used to type strains

 

FLAGELLA: (p 62) element of motility, spins to move. Flagella cannot be seen unless coated with dye.

             filament is a polymer of flagellin (H antigen), hook at proximal end, attached via basal body to bacterium:

             Gm pos: single pair of rings in basal body

             Gm neg: two pairs of rings in basal body, inner ring rotates, outer is stator            Example:

 

Styles of flagellar arrangement (more)(p63)

             peritrichous                  “around hairs”                                     E. coli, etc

             monotrichous               (single polar flagellum)                           Vibrio

             lophotrichous               [tuft hair] (2 flagellae or more at l end)   Pseudomonas

             amphitrichous              (tufts of flagellae at both ends)                Spirillum

             permit chemotaxis        attractant: steady runs, repellant: runs with many tumbles.

 

AXIAL FILAMENTS: (p 63) in spirochetes, cell wraps around axial filament

             move by boring or snake-like movement (Treponema & Leptospira)

 

MOVEMENTS: (p 64)            Run:                 clockwise rotation, 1 second (favorable conditions)

                                                Tumble:           counterclockwise, 0.1 sec. (Unfavorable conds.)

 

PILI: [hair] (p 65) hollow structures of subunits pilin [antigenic]

             1) fimbriae      aid in attachment to substrate, enhance pathogenicity, eg: Neisseria gonorrhoeae

                                       pellicle: fuzzy ort shiny layer on top of air-water interface by aerobic bacteria

             2) sex pili         aid in transfer of DNA




PROKARYOTIC ANATOMY II: CELL WALLS, PLASMA MEMBRANE, ENDOSPORES

 

6/30/83, rvsd 8 July 1995, 25 June 99, 13 July 01, 7 July 03, 14 July 04, 11 July 05, 13 July 05, 5 apr 06, 9 July 07

TFC, 7th p 77-99, Alcamo, p. 87-, Atlas, pp 111-139, TFC, 8th p 76-96, Black 6th: 90-95, Bauman 2nd,65-92

 

CELL WALL: (p 66)

20x norm pressure inside (300 psi!), cell wall hold contents in, gives structure & protection

             composed of peptidoglycan (murein), a polysaccharide with alternating NAG-NAM (10 to 65 in row):

             glycan:             NAG:   N acetylglucoseamine (p 66)

                                       NAM:  N-acetylmuramic acid, tetra peptide cross linking anchors

             cross connected via 1-5 AA (alternating D and L) with peptide cross-linking side chains .

                                       bacteria grouped according to antigenicity of cell wall (example: group A strept.)

Overview: p 68:

Gram positive     25 nm thick of peptidoglycan plus antigenic teichoic acid (p 68) (PO4 + glycerol (or ribitol) or phospholipid)

Gram negative    only 3 nm thick wall, plus second membrane on exterior of cell wall with lipopolysaccharide

                                          periplasmic space between membranes: protective enzymes

 

LIPOPOLYSACCHARIDE (LPS, in outer membrane of Gram negative bacteria) consists of:

         Lipid A (p. 68)        embedded in outer membrane, released as endotoxin on death. Trigger macrophages to release cytokines: chills, fever, weakness, generalized aches, shock, death.

         O polysaccharide   projects out, constitutes "O" antigen as in O157:H7 of pathogenic E. coli.

p 68: contrast Gm +/Gm-:           As a result of the extra barrier in Gm- bacteria, more resistant to antibiotics, salts and dyes

 

AGENTS AGAINST CELL WALLS:

         Lysozyme               (found in tears, etc) cuts polysaccharide

         Penicillin                inhibits synthesis, leads to protoplast formation in growing cells

 

Mycoplasma have no cell wall, instead have cholesterol in membrane (a unique group of bacteria)

 

PLASMA MEMBRANE: (p 69) 60% protein, 40% phospholipid, fluid mosaic model

         Mesosomes: invaginations of plasm membrane, form septum. DNA attached here incr absorb

Alcohols, quaternary ammonium compounds and polymyxins damage the plasma membrane

 

MOVEMENT ACROSS MEMBRANE: (p 71)

         Diffusion, osmosis (p 73), hypertonic solutions draw water out, cell collapses (plasmolysis)

         Facilitated diffusion, active transport (p 74) semipermeable membrane video

 

CYTOPLASM: Inclusions (stored nutrients), classification of which aid in classification:

         volutin:                       stored poly P04 (red with methylene blue, diagnostic of C.orynebacterium diphtheria)

                                             metachromatic with methylene blue staining (variable staining)

         polysaccharide            starch or glycogen, iodine shows as black/purple grains

         lipid:                          poly β-hydroxybutyric acid, sudan dyes shows (p 75)

 

ENDOSPORES: (p 547)    specialized resting cells, survive extreme heat , toxins, chemicals, xeroduric.

                                             Survives 10,000 yr, possibly 25 mil in amber?

       Four genera, only Bacillus and Clostridium are pathogenic

 

Illustrate process of sporulation:(p. 76)

                        1. spore septum forms by invagination of plasma membrane

2. newly synth DNA surrounded (forespore) by double layer of membrane (spore and cell memb)

3. Cortex of peptidoglycan between two membranes

4. spore coat of protein synthesized on outside.

Layers: 1) spore coat, 2) outer memb, 3) cortex, 4) inner memb, 5) inside: dipicolinic acid, stabilizes DNA

 

EUCARYOTIC CELLS [Not covered?]

 

Flagella and cilia

 

Cell wall

 

Plasma membrane

 

Cytoplasm

 

ORGANELLES: Nucleus, endoplasmic reticulum, golgi complex, mitochondria,

 

Centrosome and centrioles, chloroplasts, inclusions