Experiment 2 routine cultivation of bacteria The cultivation of bacteria (1)
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Experiment 2
routine cultivation of bacteria
The cultivation of bacteria (1)
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Contents1. Media for routine cultivation of bacteria
2. Techniques for bacterial cultivation and
inoculation
3. Growth characteristics of bacteria on/in different
media
4. Metabolic activities of bacteria
5. The physical effect on bacteria and commonly
used sterilizers
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1. Media for routine cultivation of bacteria
Media--- The basic ingredients of media
include carbon source 、 nitrogen source、 inorganic salt 、 growth factor and
distilled water etc.
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Classification of culture media
According to function, in general, media can be classified five types:
1. basal medium ( agar plate medium) 2. nutrient medium (blood agar plate medium) 3. selective medium 4. differential medium 5. anaerobic medium.
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According to physical properties, media can be classified three types:
1. solid medium ( 1.5~ 2.5% of agar)
agar plate /slant medium
2. semi-solid medium (0.3~ 0.5% of agar)
3. liquid medium
1 2 31
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2. Techniques for bacterial inoculation
Bacterial inoculation is a basic technique of microbiology. The operation must be under sterile condition.
1) isolation cultivation
2) pure cultivation
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• 1) Isolation cultivation• streak a bacterial culture on a nutrient agar
plate to separate the individual bacterial cells, each isolated cell will develop into a pure colony after incubation.
• continuous streak method
• quadrant streak dilution methodmethod
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2) Pure cultivation transfer a single colony from agar plate or a
pure slant culture to liquid medium, slant or semisolid medium to get a pure culture.
subculture into liquid medium
subculture into agar slant/plate
subculture into agar deep tube (semi-solid)
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3. Phenomena of bacterial growth
on agar plate (on blood agar plate)
in liquid medium
in semi-solid medium
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1) on agar plate
observe the isolated colony on the surface of nutrient agar plate; note size, shape, elevation, margin, surface, transparency and pigment (color of colony).
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on agar plate -- colonycolony
Note:
Size 大小
Shape 形状
Elevation 突起
Margin 边缘
Surface 表面
Transparency 透明度
Pigment 色素
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on blood agar plate
observe hemolysis (complete and incomplete)
phenomena around colony.
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3) in liquid medium
three growth phenomena will be seen: surface growth pellicle 表面生长 uniformly turbid 均匀混浊生长 sediment in bottom 沉淀生长
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In liquid mediumsurface growth pellicle uniformly turbid sediment in bottom
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4) in semi-solid medium nonmotile organism
motile organism
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4. Metabolic Products and Biochemical Testing
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Carbohydrate Fermentation Test
溴甲酚紫【 pH 5( 黄 ) - pH7( 紫 ) 】
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Methyl Red (MR) Test
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Voges-Proskauer (VP) Test
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Indole Test
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• Citrate utilization test
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I M Vi C test E.coli + + - - E.Aerogenes - - + +
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Hydrogen Sulfide (H2S ) Production
sulfur amino acid
sulfur inorganic compoundH2S
Pb2+
Fe2+
PbS
FeS( black )
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Urease Test
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5. The physical effect on bacteria and
commonly used sterilizers 1) Physical factor:
dry 、 moist 、 ultraviolet 、 ultrasonic wave 2) Commonly used sterilizers:
autoclave
dry-air oven
bacteriological filters
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Experiment manipulation
1. Continuous streak method or quadrant streak method strain: S.aureus or E.coli
one piece of plate/person
2. Observe phenomena of bacterial growth on agar plate in liquid medium in semi-solid medium
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Experiment report1. Continuous streak method or quadrant streak
method:
principle
procedure
result: describe the characteristics of colony
analysis
2. Draw phenomena of bacterial growth