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Growth & Culture of Bacteria

Binary Division
1 to 2 to 4 to 8 to ?

Phases of Growth
Lag
Adapt to nutrients

Log
Active growth

Stationary
Death = Growth rate

Death
Nutrients consumed
pH too low (why?)

Optimize curves in production

Log Growth

Measuring Growth
Direct Counts
Petroff-Hauser Chamber
Pros vs. Cons

Serial Dilution
10-fold serial dilutions
Pros vs. Cons

MPN (Most Probable Number)


Put 10, 1, and 0.1 ml into 10-mls broth
Repeat 5 times per volume

Statistical accurate sampling


Public Health Standards are written for MPN

Measuring Growth-contd
Turbidity
Spectrophotometer
Scale
%Transmittance
Optical Density or Absorbance

Filtration
0.45 - 0.2 um sizes
Grid Pattern on Filter
Standards for Public Health
0 E.coli / 100 ml of water

Also used for sterilization

Physical Factors: -philes


pH
Acido-, Neutro-, Alkaliphiles
Canned foods-Hot Pack vs. Pressure Cooking
High Acids- tomatoes
Neutro-corn, beans, meat

Temperature
Psychro-, Meso- Thermophiles
Growth Curve
Problems

Note the drop-off as you get closer to the Optimum!

What effect would have a fever have on Mesophiles??

Temperature Problems
Thermophiles
Hot tubs-public
Heat Resistant Enzymes in PCR

Psychrophiles
Cold food (growth in transit)
Listeria grows in hot dogs

Salt Concentration
Halophiles
Great Salt Lake
Salt Pork

Physical Factors: contd


Moisture
Beef Jerky
Lyophilization (Freeze Dry)

Oxygen
Aerboes vs. Anaerobes + ------ + ----------

Osmotic Pressure
Pseudomonas grows in distilled water
Jelly- Jams (high sugar)
Salt Pork (high salt)

Nutritional Factors

Carbon Source

Auto- or Hetero CO2 or Organic compounds

Nitrogen Source
NO3- ==> NO2 ==> NH3 to Amino Acids ==>
Proteins

Sulfur
Phosphorus
Trace Elements
Fe, some required
Host Sideophores bind Fe

More -trophs
Auxotrophs
Can live on simple compounds
NO3-, Glucose
Make all Amino Acids, Vitamins, etc.

Prototrophs
Require complex compounds
Vitamins, Amino Acids
Fastidious
Most Pathogens

Types of Media
Complex Medium
Peptones, Yeast Extract
Fastidious
Prototrophs

Synthetic Medium
Minimal Media
Auxotrophs

Exoenzymes

Secreted into environment


Catabolize food products
Protease, DNAase, Amylase, Lipase
Used in identification

Oxygen Requirments
Classification

Aerobic
Anaerobic
Falcultative Anaerobic
Microaerophilic

Outline for Topic


Media
Cultivation
Oxygen Toxicity

Flluid Thioglycollate Medium


(FTM)

Peptones, Yeast Extracts, Glucose


Thioglycollate-binds O2
Agar-slow diffusion of O2
Rezaruin- O2 indicator

FTM Media

Growth Chambers
Candle Jars
Microaerophillic
Gas-Paks
Anaerobes & Faculatative Anaerobes

Why Anerobes vs. Aerobes


O2 is reactive-toxic (binds e-s)
Requires 4 e-s to form water
Intermediates are Free Radicals

Free Radicals steal e-s


Oxidization of compounds
Loss of energy from compounds
DNA molecule breaks