Culture media

Culture media introduction, reasons, nutrients required for culture media, classification with description part covered

1. 1
culture media
Prestented by,
Pr. Namdeo Shinde
Assistant Professor
Department of Pharmaceutics,
Satara College of Pharmacy, Satara-415004
India

2. •CULTURE :
Isthetermgiventomicroorganismsthatare
cultivatedinthelabforthepurposeof
identifyingandstudyingthem.
•MEDIUM:
Isthetermgiventothecombinationof
ingredientsthatwillsupportthegrowthand
cultivationofmicroorganismsbyprovidingall
theessentialnutrientsrequiredforthegrowth
(i.e.multiplication)inordertocultivatethese
microorganismsinlargenumbertostudy
them.
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CULTURE AND THE MEDIUM

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Thefoodmaterialorsubstancesrequiredforgrowing
microorganismsinvitro(outsidethebody)iscalled
culturemedium.
Reasonsforbacterialcultural:
-subsequent clinical diagnosis.
-studying its morphology and its identification.
-Bacteria have to be cultured in order to obtain antigens
from developing serological assay for vaccines.
-Certain genetic studies and manipulations of the cells
also need that bacteria be cultured in vitro.
-Culturing on solid media is another convenient way of
separating bacteria in mixture

4. •Microbiologicalculture:whichare
usedforgrowingmicroorganisms,
suchasbacteriaoryeast.
•Themostcommongrowthmediafor
microorganismsarenutrientbrothsandagar
plates
•Specializedmediaaresometimesrequiredfor
microorganismandcellculturegrowth.
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5. CULTURE MEDIA
•Used to grow bacteria
•Can be used to-
Enrich the numbers of bacteria.
Select for certain bacteria and suppress others.
Differentiate among different kinds of bacteria.
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6. NEED FOR CULTURE MEDIA
•Itisusuallyessentialtoobtainacultureby
growingtheorganisminanartificialmedium.
•Ifmorethanonespeciesortypeoforganismare
presenteachrequirestobecarefullyseparatedor
isolatedinpureculture.
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7. BASIC REQUIREMENTS OF CULTURE MEDIA
•NUTRIENTS :
Energy source
-Phototrophs-light-Rhodospirillumrubrum
-Chemotrophs-chemical energy-E. Coli
Carbon source:CO2, organic complounds
Nitrogen source-atm./nitrate,nitrite
•MINERAL SALTS :
Sulphates, phosphates, chlorides and carbonates of K,
Mg and Ca
A suitable pH-7.2-7.4
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8. BACTERIAL GROWTH FACTORS
ARGININE E.COLI
GLUTATHIONE GONOCOCCI
CHOLESTEROL MYCOPLASMA
ARYL SULPHATE, AMIDE ATYPICAL MYCOBACTERIA
GLYCEROL MYCOPLASMA HOMINIS
SULFONAMIDES RIKETTSIA
TRYPTOPHAN SALMONELLA TYPHI
L-CYSTEINE LISTERIA MONOCYTOGENS
SODIUM CHLORIDE VIBRIO PARAHAEMOLYTICUS
FACTOR X & V H.INFLUENZAE
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12. CLASSIFICATION
BASED ON
PHYSICAL STATE
BASED ON
PRESENCE OF
MOLECULAR
OXYGEN AND
REDUCING
SUBSTANCES
BASED ON
NUTRITIONAL
FACTORS
LIQUID MEDIA AEROBIC MEDIA SIMPLE MEDIA
SEMISOLID MEDIA ANAEROBIC MEDIA COMPLEX MEDIA
SOLID MEDIA SYNTHETIC MEDIA
SPECIAL MEDIA
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13. SPECIAL MEDIA
A. ENRICHED MEDIA
B. ENRICHMENT MEDIA
C. SELECTIVEMEDIA
D. DIFFERENTIAL MEDIA
E. INDICATORMEDIA
F. TRANSPORT MEDIA
G. SUGAR MEDIA
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14. SIMPLE MEDIA
Simple or basal media are culture media which contain the minimum adequate
nutrition for non fastidious organisms
Fastidious organisms: Only grow when specific nutrients are included in its diet.
Example:-Nutrientbroth/agar
Peptone water
Composition:-
Lab-Lemco -10gm
peptone-10gm
NaCl-5gm
Distilled water-1000ml
-When 2-3% agar is added ,then we have it as nutrient agar.
-For semisolid media –agar concentration is 0.2-0.4%
Uses:-
1.Thisisbasisofmostofthemediausedinthestudyatcommon
pathogenicbacteria.
2.Itisusedforsubculturesofcertainorganisms.

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16. NUTRIENT BROTH
SEMISOLID
NUTRIENTAGAR
SOLID
LIQUID MEDIA

17. PEPTONE WATER
•TYPE : Basic liquid media
•APPEARANCE : clear, colorless, watery, usually
in test tube
•Composition :
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PEPTONE 10 g
SODIUM CHLORIDE, NaCl 5 g
WATER 1 litre

18. USES OF PEPTONE WATER:
Themediaisusedchieflyasthebasisfor
carbohydrate fermentation media.
NUTRIENTBROTHS:
maycontainasmallamountofsugarderived
frommeatanditisessentialthatthebasal
mediumtowhichvariouscarbohydratesare
addedforfermentationtestsshouldbefree
fromnaturalsugars.
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19. Itisalsousedtotesttheformationofindole.
Cultureoforganismsfordemonstrationofmotility
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NUTRIENTBROTHS:

20. COMPLEX MEDIA
Complex media have added ingredients for bringing out certain properties or
providing special nutrients required for growth of the bacteria.
SYNTHETIC MEDIA
These are prepared from pure chemicals and the exact compositions of
medium is very well known.
Example :-Dubo’s medium
SEMIDEFINED MEDIA
Inthesemediatheexactchemicalcompositionoftheconstituentsisnot
knownbecausesubstanceslikemeatandpeptoneareused.
Mostoftheculturemediausedforroutinediagnosticworkaresemidefined
culturemedia.

21. SPECIAL MEDIUM
• ENRICHED MEDIA
When basal medium is added with some nutrients such as blood,
serum or egg is called enriched media.
They are used to grow bacteria which are more exacting in their
nutritional needs.
Examples:-
Dorset’sEggMedium.
It is a creamy coloured opaque
slope kept in screw copped bottle
Selective medium for isolation
of Mycobacterium tuberculosis.
Composition: Hen’s egg, Nutrient broth

22. BLOOD AGAR
•TYPE : Enriched media.
•APPEARANCE : Red color.
•COMPOSITION :
Sterile Nutrient agar + Defibrinated sheep blood
USES :
Routine culture
Widely used in medical bacteriology
It is also an indicator medium showing the haemolytic
properties of bacteria such as Streptococcus pyogenes.
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23. CHOCOLATE AGAR
Also called Heated blood agar.
•TYPE: Enriched media.
•APPEARANCE : Chocolate brown color.
PROCEDURE:
Melt the desired amount of nutrient agar.
Cool it in a water –bath at 75º C .
Add 10 ml of sterile blood .
Allow the medium to remain at 75º C.
Mixing the bloodand agar by gentle agitation from time
to time until the blood become chocolate brown in
color, within about 10 min. Then pour in plates.
USES: CULTURE OF Neisseria, Haemophilus
influenzae, Pneumococcus
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24. ENRICHMENT MEDIA
Inthismedia,ithasastimulatingeffectonthebacteriatobegrownor
inhibitsitscompetitors.
Thisresultinanabsoluteincreaseinthenumberofwantedbacteria
relatedtootherbacteria.
Example:-SeleniteFbroth
ItisenrichmentmediumforcultureofSalmonellatyphiandparatyphi
bacillifromstoolsample
Principle:-
AtneutralpHsolutionacidsalinityhashightoxicitytocoliformgroupof
bacteriaandnottomostofthesalmonellagroups.

25. SELENITE F BROTH Alkaline peptone water

26. SELECTIVE MEDIA
Itisamediuminwhichcertainsubstancesarepresentwhichinhibitallother
bacteriaexceptthedesiredbacteria.
It encourages the growth of particular species from a mixed inoculum.
Example:-TCBS
-It is light green translucent medium kept in petridish
-It is selective medium for Vibriocholera
-Principle:-
Bile salt inhibit the growth of normal
commensals(unwanted bacteria).
Vibriochloeraeproduce acid by fermentation
of sucrose which acts on bromothymolblue
(indicator) producing yellow colonies.

27. Salmonella-Shigella agar plate (SS)Salmonella Shigella Agar with DCA

28. MAC CONKEY AGAR
•MacConkey agaris a culture medium
designed to grow Gram-negative bacteria. It is
a useful medium for the cultivation of
enterobacteriacea.
•It shows lactose fermenter as red colonies
while non-lactose fermentor as white/pale
colonies.
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DIFFERENTIAL MEDIA

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MacConkeyagar
showing both
lactose and non-
lactose fermenting
colonies.
Lactose fermenting
colonies are pink
whereas non-
lactose fermenting
ones are colourless
or appear same as
the medium.

30. •Gram-negativebacteriagrowingonthemediaare
differentiatedbytheirabilitytofermentthesugar
lactose.
•LactosefermentercausethepHtodropandis
detectedbyneutralred,(redatpH'sbelow6.8.)which
appearasbrightpinktoredcoloniesontheagar.
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31. •Uses
•ActingasavisualpHindicator,theagar
distinguishesthoseGram-negativebacteria
thatcanfermentthesugarlactose(Lac+)from
thosethatcannot(Lac-).
Thismediumisalsoknownasan
•"indicatormedium"
•"lowselectivemedium".
•Absenceofelectrolytesservestoinhibit
swarmingbyProteusspecies
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32. Lac+
•Byutilizingthelactoseavailableinthe
medium,Lac+bacteriasuchas
Escherichiacoli
Enterobacterspp.
Klebsiellaspp.
willproduceacid,whichlowersthepHofthe
agarbelow6.8andresultsintheappearance
ofred/pinkcolonies
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33. CLED
(Cystine Lactose Electrolyte Deficient medium)
•Itisavaluablenon-inhibitorygrowthmedium
usedintheisolationanddifferentiationof
urinaryorganisms.
•Beingelectrolytedeficient,itpreventsthe
swarmingofProteusspecies
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Description:
Lactose&non
lactosefermenters
onCLEDmedium.

35. INDICATOR MEDIUM
Thesemediacontainanindicatorwhichchangescolourwhenbacteria
growonthem.
Example:-WilsonandBlairmedium
ForisolationofSalmonellatyphiandS.paratyphi
Theyappearasblackcolonies
Principle:-Theblackcolourofcoloniesisduetotheabilityofthese
organismstoreducebismuthsulphitetosulphideinthepresenceof
glucosecoliformsareinhibitedbybrilliantgreenandbismuthsullphite

36. TRANSPORT MEDIUM
Theseareusedforthetemporarystorageof
specimensbeingtransportedtothelaboratoryfor
cultivation.
Suchmediaideallymaintaintheviabilityofall
organismsinthespecimenwithoutalteringtheir
concentration.
Transportmediatypicallycontainonlybuffersand
salt.
Thelackofcarbon,nitrogen,andorganicgrowth
factorspreventsmicrobialmultiplication.
Transportmediausedintheisolationofanaerobes
mustbefreeofmolecularoxygen.
STUART TRANSPORT BROTH

37. CHARACTERISTICS OF TRANSPORT MEDIA:
•It should be non-toxic
•It should not promote or inhibit the bacterial growth
•It should be easy to carry and transport
Examples:
1.Venkatraman Ramakrishnan medium
2.Buffered glycerol saline transport medium
3.Cary and Blair medium
Cary and Blair medium

38. ANAEROBIC MEDIUM
These media are used to grow anaerobic organisms.
Examples:-
Thioglycollate broth
Robertsons Cooked Meat Medium

39. •Mueller-Hintonagarisan
microbiologicalgrowth
mediumthatiscommonly
usedforantibiotic
susceptibilitytesting.
•Originallyformulatedfor
isolationofNeisseria
species.
•Itisalsousedtoisolate
andmaintainNeisseria
andMoraxellaspecies.
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Mueller-Hinton agar

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