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Microbe Report 2

Microbe Report 2

Ratings: (0)|Views: 11,921|Likes:
Published by maibma
1. EXAMINATION OF CULTUER FOR MOTILITY
2. CULTURE TECHNIQUE
1. EXAMINATION OF CULTUER FOR MOTILITY
2. CULTURE TECHNIQUE

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Categories:Types, Research, Science
Published by: maibma on Sep 30, 2009
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12/28/2012

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PRACTICAL 2I.EXAMINATION OF CULTURES FOR MOTILITYIntroduction:
The motility of bacteria is determined by the presence of flagella. Flagella are a longthread like structures. We can classify the organism by their motility. There are two types of determining motility, hanging drop method and soft agar stab
Objective
 –To identify and differentiate between true motility and Brownian motion –To familiarize and perfect students aseptic techniques in handling aseptic transfers andisolating a pure culture.
MaterialsPer pair of students
4 soft agar bijous2 cavity slides2 wire loops2 straight wiresCover slips
Per bench
1 broth culture of 
 Pseudomonas aerugenosa
1 broth culture of 
Staphylococcus aureus
Plasticine
Procedure(A)Hanging Drop preparation
1.
A tiny amount of plasticine was placed at each corner of the clean cover slip
2.
One or two loopfuls of the broth culture was placed in the centre of the cover slip.
3.
Concave area of the cavity was placed over the centre of the cover slip. The slip was press down gently and was inverted.4.Under the low power objective and reducing light, the edge of the drop was centred5.The movement of cells was observed. The movement of true motility and Brownianmotion was differentiated.
(A)Soft agar ‘stab’
1.A straight wire was dip into the broth culture by using antiseptic technique.
2.
One single stab was made in the centre of the soft agar down to the bottom.
3.
The soft agar was not disturbed, shacked and was placed upright for incubation
 
I.CULTURE TECHNIQUEIntroduction:
As most microorganisms are potentially pathogenic it is essential that they be handled by procedures which avoid contamination of the microbiologist. Furthermore, since microorganismsare so ubiquitous it is necessary to avoid contamination of microbial culture by stray microorganisms.
Objective
 –To become familiar with the basic movements of aseptic culture technique and our owntechnique.
1) Inoculums transfer(A)Technique of transferring culture from one agar plate to another bystreakingMaterials
1 wire loopPlate culture of 
 Escherichia coli
1 nutrient agar plateBunsen burner 
ProcedureRemember that the agar is a fragile medium and only gentile pressure with the oop isrequired.
1.The Bunsen burner was lighted and the wire loop was holed by right hand.
2.
The wire loop is then sterilized by flaming it in the Bunsen burner until it is red heat. Thewire loop was allowed for 5 seconds to cool it down.
3.
The lid of Petri dish containing
 E.coli
was opened by using left hand. Then, small loopwas removed from the inoculums and the lid was closed immediately.
4.
The second Petri dish was opened with left hand and the inoculum was spread at the smallarea at the top edge of the agar and spread it over a small area (primary inoculums). Thenthe loop was flamed.5.After the loop was cooled make four or five single strokes starting from the primaryinoculums. The loop was flamed again and overlapping series of stroke covering anther area was made until all the surface of the agar was covered.
6.
The plate was closed and was placed inverted for incubation at 37’C for 24-48 hours.
(A)Technique of transferring culture from an agar plate to a bottle of nutrientbroth
 
Materials
1 wire loopPlate culture of 
 Escherichia coli
1 bottle of nutrient brothBunsen burner 
Procedure
1.
The wire loop was sterilized and small loop was removed from the inoculum in the plate(as in 1) a) 3.)
2.
The bottle was taking by using left hand and twist the cover off the bottle by using right hand(while holding the wire loop) and the top of the bottle was flamed evenly near the Bunsen burner.3.With the right hand, the loop was dip in to the nutrient broth and was mix gently withminimum movement.4.The bottle was flamed as in step 3 and its cap was screwed back.
5.
The loop was carefully flamed by placing in the cooler part of the Bunsen flame andgradually drawn to the hotter region of the flame. (to reduce amount of splattering).6.The bottle was labelled and incubated at 37’C for 24-48 hours
(A)Isolating a pure culture from a mixed cultureMaterials
A plate of mixed culture of 
 Bacillus subtilis
and
Staphylococcus aureus
1 wire loopA plate of nutrient agar 1 bottle of nutrient broth
Procedure
1.
The mix culture was streaked on the nutrient agar plate to obtained a pure culture of 
 Bacillus subtilis
and
Staphylococcus aureus
2.
The mixed culture was transferred into the bottle of nnutrient broth3.The bottle and plate was labelled and incubated at 37’C for 24-48 hours.
Discussion:1. EXAMINATION OF CULTUER FOR MOTILITY 

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