Professional Documents
Culture Documents
Week 1 Lecture
Week 1 Lecture
iv. Inclusions
BACTERIAL PHYSIOLOGY, • Metachromatic Granules – represents reserves of
polyphosphates used in the synthesis of ATP.
METABOLISM, GENETICS, GROWTH • Polysaccharide Granules - consist of glycogen and
starch granules
AND NUTRITION, AND BACTERIAL v. Spores
• highly refractile resting cells that are highly durable
IDENTIFICATION and dehydrated with thick walls
B. CELL ENVELOPE
i. Plasma Membrane (Cell
BACTERIAL PHYSIOLOGY
ii. Cell Wall
iii. Periplasmic space
PROKARYOTIC CELL STRUCTURES
iv. Outer Membrane
• Prokaryotic Cell Components
B. Cell Envelope
A. Cytoplasmic Structures
i. Plasma Membrane (Cell Membrane)
B. Cell Envelope Structures
• A phospholipid bilayer with embedded proteins that
C. Surface Polymers or Appendages
envelop the cytoplasm.
A. CYTOPLASMIC STRUCTURES
i. nuclear Area (nucleoid) ii. Cell Wall (Peptidoglycan or murein layer)
ii. plasmid Maintains the shape of the cell and protects cell
iii. ribosomes from osmotic pressure
iv. inclusions Repeating disaccharide attached by
v. Endospore polypeptides
A. CYTOPLASMIC STRUCTURES
i. Nuclear Area (Nucleoid)
• single circular chromosome attached to a
mesosome
ii. Plasmids
• small circular, dsDNA molecule iii. Periplasmic space
• antibiotic resistance Between cell membrane and cell wall.
iii. Ribosomes
Consist of gel like matrix containing nutrient
• site of protein synthesis
binding nutrients
• consists of RNA and protein
Contains enzymes for degrading and detoxifying
macromolecules
CELL ENVELOPE
CHARACTERISTIC GRAM + GRAM -
Peptidoglycan Thick Absent
layer (multilayere)
Teichoic acid Present in many Absent
INCUBATION
- CO2 incubator is not BACTERIAL IDENTIFICATION
available, used candle gar or Conventional Methods
Gaspak jar with CO2 - Biochemical characterization serves as the
generator fundamental source of information for ID of
most bacteria after microscopy.
REPORTS • Motility test - wet mount and hanging drop preparation
Save culture until satisfactory growth is • Staining - Gram staining, acid-fast staining, structural
obtained and final diagnosis is made. staining
Some cultures made be held for several weeks • Manual biochemical test - carbohydrate fermentation
before final report is made (LIA), catalase and coagulase tests
o Blood culture - 7-10 days • Use of routing and selective media
o Brucella and M. tb - 4 weeks
o L. monocytogenes - 4 months Analytical Profile Index
All media except thioglycolate broth should be - Consists of plastic strips and microtubes that
stored in the refrigerator . contain dehydrated biochemical substrates
The number of media stored should not exceed • Commonly used for gram (-) enteric bacteria
2-3-week supply. • Same principle with biochemical manual tubed method
All media must be checked before use. • Principle: Biochemical substrates (pH-based substrates)
are inoculated with pure culture suspended in sterile
physiologic saline and incubated for 18-24 hrs at 35C, some
reagents may be added after incubation.
STAINING TECHNIQUES
Types of Stains
Simple Stains
Indirect, Relief, or Negative Stains
Special Stains - Capsular Stain, Cell Wall Stain,
Metachromatic Granules, Flagellar
Differential Stain