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Bio 127 Lecture 4b Culture Media

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78 views19 pages

Bio 127 Lecture 4b Culture Media

For lecture class

Uploaded by

cheramae ances
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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LABORATORY CULTURE OF

MICROORGANISMS
 Microorganisms can be grown in the
laboratory in culture media
containing the nutrients they
require.
 Successful cultivation and
maintenance of pure cultures of
microorganisms can be done only if
aseptic technique is practiced to
prevent contamination by other
microorganisms.
CULTURE MEDIA: TYPES

Media can be classified on three primary levels


1. Physical State
2. Chemical Composition
3. Functional Type
CULTURE MEDIA: COMPOSITION
 Culture media supply the nutritional
needs of microorganisms ( C ,N, Phosphorus,
trace elements, etc)
 defined medium : precise amounts of highly
purified chemicals
 complex medium : highly nutritious
substances.
 In clinical microbiology,
 Selective : contains compounds that selectively
inhibit
 Differential: contains indicator
LIQUID MEDIA
 Water-based solutions
 Do not solidify at temperatures above freezing /
tend to be free flowing
 Includes broths, milks, and infusions

 Measure turbidity

 Example: Nutrient Broth, Lactose Broth


SEMI-SOLID MEDIA
 Exhibits a clot-like consistency at ordinary room
temperature

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 Determines motility

 Used to localize a reaction at a specific site.

 Example: SIM for hydrogen sulfide production


and indole reaction and motility test.
SOLID MEDIA

 Firm surface for discrete colony growth


 Advantageous for isolating and culturing

 Two Types

1. Liquefiable (Reversible)
2. Non-liquefiable
 Examples: Gelatin and Agar (Liquefiable)
Cooked Meat Media,
Potato Slices (Non-liquefiable)
SELECTIVE MEDIA

 Contains one or more agents that inhibit the


growth of a certain microbe and thereby
encourages, or selects, a specific microbe.
 Example: Mannitol Salt Agar [MSA] encourages
the growth of S. aureus. MSA contain 7.5% NaCl
which inhibit the growth of other Gram positive
bacteria
DIFFERENTIAL MEDIA
 Differential shows up as visible changes or
variations in colony size or color, in media

9
color changes, or in the formation of gas
bubbles and precipitates.
 Example: Spirit Blue Agar to detect the
digestion of fats by lipase enzyme. Positive
digestion (hydrolysis) is indicated by the dark
blue color that develops in the colonies. Blood
agar for hemolysis (α,β,and γ hemolysis), EMB,
MacConkey Agar, …etc.
10
ENRICHMENT MEDIA

 Is used to encourage the growth of a particular


microorganism in a mixed culture.
 Blood agar , chocolate agar
METHODS USED TO MEASURE MICROBIAL
GROWTH

 Count colonies on plate or filter (counts


live cells)
 Microscopic counts

 Flow cytometry (FACS)

 Turbitity
VIABLE COUNTS

 Each colony on plate or filter arises from single live cell


 Only counting live cells
Direct Count
Pour Plate

16
17
Direct Count
Spread or
Streak Plate

18
MICROSCOPIC COUNTS

 Need a microscope, special slides, high power


objective lens
 Typically only counting total microbe numbers,
but differential counts can also be done
TURBITITY

 Cells act like large


particles that scatter
visible light
 A spectrophotometer
sends a beam of visible
light through a culture
and measures how much
light is scattered
 Scales read in either
absorbance or %
transmission
 Measures both live and
dead cells

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