 INTRODUCTION

 CULTURE MEDIAS
 CULTURE METHODS
 Molds and yeasts are widely distributed in

air, dust, fomites and normal flora.
 Humans are relatively resistant.
 Fu...
 A)BASAL MEDIA
 1)SABOURAUDS DEXTROSE AGAR
 2)NEUTRAL SABOURAUDS DEXTROSE AGAR
 3)SDA WITH ANTIBIOTICS

 B)NUTRITIONA...
 8)CZAPEK`S DOX AGAR

 D)MEDIA FOR STIMULATION OF

ASCOSPORE OF PERFECT FUNGI
 1)ALPHACEL-YEAST EXTRACT AGAR

 2)SOIL ...
 PEPTONE








-10 gm
AGAR
-20 gm
DEXTROSE
-40 gm
DISTILLED WATER
-1000ml
Autoclave at 121*c for 15 min.
Adjust ...
 INGREDIENTS
 CORN MEAL
 AGAR
 DISTILLED WATER
 TWEEN 80

-8 gm
-4 gm
-200 ml
-2 gm
 A Heavy inoculum of yeast is streaked across a








plate containing the medium.
Cover slip is placed over it.
...
 Used for growing fastidious pathogenic fungi such










as
-Histoplasma capsulatum
-blastomyces dermatitis
 ...
 Dissolve ingredients by boiling.
 Dispense into screw capped bottles.
 Autoclave at 121*c for 15 min
 Cool in slanted...
 For cryptococcus neoformans
 Can utilise creatinine as a source of nitrogen
 Colonies are brown to black due phenoloxi...
 Niger seed ectract
 Glucose
 Chloromphenicol
 Gentamicin
 Dipheny solution

 Agar
 Distilled water
 Autoclave at ...
 Used for
 Histoplasma capsulatum
 Blastomyces dermatitidis
 Cryptococcus neoformans

 Ingredients
 Blood agar base
...
CULTURAL METHODS


Sabrourd’s medium is used world-wide and is
generally satisfactory



Some workers prefer malt peptone agar



It is ...
.

Petridish

Test tube

Surface area

Large

Small

O2 supply

Good

Poor

Security of closure

Poor

Good

Detection of ...
Blood culture
• Same as that of microbiology
• For manual system blood culture bottle with
agar and mycological broth or s...
TISSUE
Biopsy and other tissue should be reduced in
size 1 -2mm
 Put these pieces into agar medium, it should
contain ant...
SWABS

o
o

Heavily feed swab rotate over the surface of media several times
Secondary dilution strokes should be made wit...
Urine

Peritoneal fluid

 Spread 0.1ml un concentrated

 Process as urine but take an

urine over the surface of agar
co...
INCUBATION
 Most fungi and moulds grow at room temperature (25





30*c)
Some at body temperature (35-37*C)
Some are...
REFERENCE
BIRD SEED
AGAR
Culture of fungus
Culture of fungus
Culture of fungus
Culture of fungus
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Culture of fungus

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Culture of fungus

  1. 1.  INTRODUCTION  CULTURE MEDIAS  CULTURE METHODS
  2. 2.  Molds and yeasts are widely distributed in air, dust, fomites and normal flora.  Humans are relatively resistant.  Fungi are relatively nonpathogenic
  3. 3.  A)BASAL MEDIA  1)SABOURAUDS DEXTROSE AGAR  2)NEUTRAL SABOURAUDS DEXTROSE AGAR  3)SDA WITH ANTIBIOTICS  B)NUTRITIONALLY DEFICIENT MEDIA  1)CORN MEAL AGAR  2)RICE STARCH AGAR  C)ENRICHED/SELECTIVE MEDIA        1)BRAIN HEART INFUSION AGAR 2)BI PHASIC MEDIUM 3)CYSTEIN HEART AND Hb AGAR 4)BLOOD AGAR 5)BIRD SEED AGAR 6)LJ MEDIUM 7)DERMATOPHYTE TEST MEDIUM
  4. 4.  8)CZAPEK`S DOX AGAR  D)MEDIA FOR STIMULATION OF ASCOSPORE OF PERFECT FUNGI  1)ALPHACEL-YEAST EXTRACT AGAR  2)SOIL EXTRACT AGAR  E)MEDIA USED FOR BIOCHEMICAL TESTS  1)TETRAZOLIUM REDUCTION MEDIUM  2)CARBOHYDRATE FERMENTATION MEDIA  3) CARBOHYDRATE ASSIMILATION MEDIA  4)UREASE MEDIUM  5)DIAZONIUM BLUE B REACTION
  5. 5.  PEPTONE        -10 gm AGAR -20 gm DEXTROSE -40 gm DISTILLED WATER -1000ml Autoclave at 121*c for 15 min. Adjust pH to 5.5 Saprobic fungi may overgrow It obscuring real pathogen
  6. 6.  INGREDIENTS  CORN MEAL  AGAR  DISTILLED WATER  TWEEN 80 -8 gm -4 gm -200 ml -2 gm
  7. 7.  A Heavy inoculum of yeast is streaked across a       plate containing the medium. Cover slip is placed over it. Streak should project beyond cover slip. Examine under low power at edge of cover slip. It is a sort of junction of aerobic and anerobic condition. Clamidiospores are best found in this area. Shows clamidiospores seen in candida albicans after 24-48 hrs incubation at 25*c
  8. 8.  Used for growing fastidious pathogenic fungi such        as -Histoplasma capsulatum -blastomyces dermatitis  INGREDIENTS Brain heart infusion agar Glucose L cysteine hydrochloride Agar Distilled water -37 gm -20 gm -1gm -20gm -900gm
  9. 9.  Dissolve ingredients by boiling.  Dispense into screw capped bottles.  Autoclave at 121*c for 15 min  Cool in slanted position with one inch butt  pH adjusted to 6.7 Store in refrigerator
  10. 10.  For cryptococcus neoformans  Can utilise creatinine as a source of nitrogen  Colonies are brown to black due phenoloxidase produced by organism.
  11. 11.  Niger seed ectract  Glucose  Chloromphenicol  Gentamicin  Dipheny solution  Agar  Distilled water  Autoclave at 121*c for 15 min.  Dispense into plates -200ml -1 gm -400gm -25 mg -10ml -20gm -800ml
  12. 12.  Used for  Histoplasma capsulatum  Blastomyces dermatitidis  Cryptococcus neoformans  Ingredients  Blood agar base  Sheep blood  Distilled water -40gm -50ml -1000ml
  13. 13. CULTURAL METHODS
  14. 14.  Sabrourd’s medium is used world-wide and is generally satisfactory  Some workers prefer malt peptone agar  It is claimed that, in the latter from the syrup is slightly more inhibitory to bacteria and produces more rapid growth and sporulation of fungi than sabouraud’s medium. In additional it is often possible to distinguish mixed cultures of yeast species by their colony morphology on malt agar Two types of containers are used for culture 1. Petri dish 2. Culture tube 
  15. 15. . Petridish Test tube Surface area Large Small O2 supply Good Poor Security of closure Poor Good Detection of mixed culture Easy Hard
  16. 16. Blood culture • Same as that of microbiology • For manual system blood culture bottle with agar and mycological broth or sabouraud broth media may be employed • Aerat the cultures periodically by shaking and sub culture routinely rather than to wait until the medium is cloudy
  17. 17. TISSUE Biopsy and other tissue should be reduced in size 1 -2mm  Put these pieces into agar medium, it should contain antibiotics if the material is contaminated with bacteria 
  18. 18. SWABS o o Heavily feed swab rotate over the surface of media several times Secondary dilution strokes should be made with a sterile loop CSF o A loop full of spun deposit should be take to inoculate the agar media in usual manner
  19. 19. Urine Peritoneal fluid  Spread 0.1ml un concentrated  Process as urine but take an urine over the surface of agar containing antibiotics  Centrifuge the specimen and remove a loop full of sediment to another agar plate of the same medium and streak out from the well in the normal way additional sample of 1ml of the neat dialysate and spread over a plate containing mycological medium .
  20. 20. INCUBATION  Most fungi and moulds grow at room temperature (25    30*c) Some at body temperature (35-37*C) Some are dimorfhic fungi All media are incubated at 25˚ to 30˚C initially and, when a potential dimorphic organism is isolated an attempt is made to convert it to the tissue phase by subeultuning it and incubating the new set of culture at 35˚C The pathogenic fungi are aerobic organisms. A good supply of oxygen is mandatory if they are to be isolated in primary culture
  21. 21. REFERENCE
  22. 22. BIRD SEED AGAR
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