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BACTERIA CULTURING

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by

Mariam Kaleem

on 25 September 2014

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Transcript of BACTERIA CULTURING

MICROBIOLOGICAL CULTURES
A culture media is a solid/ semi-solid or liquid preparation used to grow, transport and store microbes.

Agar plate


Agar slant




Agar deep tube


Chemical types of culture media
Defined or Synthetic media
WHAT ARE PIPETTES?
End of Lecture
Protocol Provided
Follow Instructions
Ask questions
Disinfect Tables and throw bio hazards waste in large bins (correct ones ) by incubator


Reminder: Quiz Next Week
Cover Lectures: PowerPoint and things Mentioned in Class (notes?
Things Needed for Lab today
Solid Media
Semi- Solid
much like any other agar media except that a smaller percentage of agar is present making the agar much softer
Liquid Media
The major difference of the 3, is the use of agar - "a solidifying agent" and how much is used
Minimal Media
Auxotrophs

Complex Media
Peptones, Yeast Extract
Fastidious
Prototrophs
Inoculating Needles and loops
Pure culture (stock culture) is a growth of a single species of microbe
Subculturing is transfering inoculum from a stock culture to fresh media
Pure culture (stock culture) is a growth of a single species of microbe

Mixed Culture is a growth of multiple species of microbe

Today's Experiments Techniques: 1. Streak Plate Technique
Isolates single colonies (usually mixed colonies) by streaking
SERIAL DILUTION- For Spread Plate and Pour plate

Volume Transferred
Final Volume
X
dilution factor of initial
Volume Transferred (Each)
Final Volume: 9 + 1 = 10ml
Dilution factor of the diluted Tube
Each dot represents a colony.

A bacterial colony is defined as a visible cluster of bacteria growing on the surface of or within a solid medium, presumably cultured from a single cell.

Colonies are supposed to be counted.

A viable number of colonies is between
25-250

Find the number of Colony Forming Unit/ ml :
Number of colonies from a plate
X
1
Dilution factor of the diluted Tube
Example
Initial tube/ stock culture is always
=10^0
Dilution # = 1
for tube
10
X
10^0
= 10^-1
Dilution # = 1
for Tube
10
x
10^-1
=1
10
x
10^-2
=10^-3
=10^-2
# of colonies Plate 1= 30
# of colonies '
plate 2 =3
Appropriate for cfu/ ml= 30 X 1/10^-2
= 3X 10 ^3 cfu/ml
2.Spread Plate Technique
Low cell count cultures are spread of the surface of a agar petri dish using either an L-shaped glass rode, or a plastic triangular spreader or a hockey stick until the culture has been absorbed by the agar
Bacterial cultures
Inoculating loops
Spreader
Test Tubes
with salines
Agar plates
Micro pipettes
Gloves
Pour Plate Technique

It can be used to determine the number of microbes/mL or microbes/gram in a specimen. It also used to study the motility of the bacteria It has the advantage of not requiring previously prepared plates, and is often used to assay bacterial contamination of foodstuffs.

Aliquot of bacteria unto plate- diluted or undiluted
Melted plate count agar, 45 C is added to sample and mixed
The streak plate method is a rapid qualitative isolation method.
It is essentially a dilution technique that involves spreading a loopful of culture over the surface of an agar plate.
3 Physical Properties
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