isolation of fungi for microbiology undergraduate .ppt

isolation of fungi

1. Isolation of fungi
Assist.Prof.: Nareen Q. Faqi Abdullah

2. Fungicanbefoundinmanydifferentenvironment
Theywillgrowonalmostanything
Theycanbefoundinforestsandgardens;theycan
alsogrowonourbody
Manyspeciesareoftenfoundonfoodsintheformof
yeastsandmolds
Fungifoundinsoil(especiallycontainingorganic
matter),waterandeveninair

3. Theyhaveimportantroleindecomposing
andrecyclingofcompoundsubstancesin
ecosystem
Fungiisolatefromitsnaturalhabitatand
culturedonselectivemediainordertoidentify
thefungalspeciesdependingonits
morphologyofcolonyandcharactersof
spores

4. Collection, preparation and cultivation of fungal samples
isolated from:
1.Air
ByopeningPetridishthatcontainssterilizedPDAinlaboratory
airforabout5minutes
Thenclosed
Incubatesat25
o
Cfor5-7days

5. 2. Water
a. Tap water:
1mloftapwaterpourintoaPetridishcontainssterilizedPDA
Incubatesat25
o
Cfor5-7days
b.Sewagewater:
Thefungalsporeswhichwasfoundinsewagewatercanbe
isolatebypouring1mlofsewagewaterintoaPetridish
containssterilizedPDA
Thenincubatesat25
o
Cfor5-7days

6. 3. Soil:
a.Directly:
1gmofsoiltakesrandomlyfrom5differentplaces
MixedwellthenspreadsomeofsoilintoaPetridishcontainssterilePDA
Incubateat25
o
Cfor5-7days
b.Indirectly:
Soilfungicanbeisolatedbyserialdilution,forthispurpose1gmofsoil
transferto9mlofSDW
Aftershakingwell1mlofthissuspensiontransfertoatubecontaining9ml
SDWbyusingsterilepipette
Shakingwellserialdilutioncontinuedto10
-6
Then1mlofeachdilutiontransfertoaPetridishthatcontainssterilePDA
Thenincubateat25
o
Cfor5-7days

8. 4. Infected human:
The infected area cleans with 70% ethanol, and then the
specimens were collect as the followings:
1. Skinscales:
Collectbyscrapingthesurfaceofthemarginofthelesion
byusingacleanslide
2. Crusts:
Collected by removing part it on healthy skin, by using
sterilized scissors or forceps
3. Nail pieces:
Collect by taking a part of the infected nail by using
sterilized scissor
4. Hairs:
Collect by removing broken hairs from the margin of the
lesion using sterilized forceps

9. 5. Conjunctivalswabs:
Withsterileswabs,theconjunctivalswabsaretakenfromthe
conjunctivaofpatientwithconjunctivitisinsterilesaline
6. Ear swabs:
Withsterileswabs,theearswabsaretakenfromtheearcanalof
diseasedmanoranimalsinsterilesaline

10. 7. Nasal swabs:
Withsterileswabs,thenasalswabsweretakenfrom
thenasalcavityofdiseasedmanoranimalsinsterile
saline
8. Blood samples:
Bloodsamplesareasepticallycollectedintubes
containinganticoagulant.Thesiteisswabbedwith
alcoholthenitispuncturedwithasterileneedle
attachedtothetubeandanamountof2mlareallowed
topasstothetube

11. 9. Faecalsamples:
Faecalsamplesarecollectedinsterileplasticbags.
Thesamplesarepreparedbymixingsampleinsterile
normalsaline
10. Oral swabs:
Withsterileswabs,theoralswabsweretakenfromthe
siteofinfectioninsterilesaline

12. These samples collect as above then:
1. Direct microscopic examination:
Partofthespecimenusedfordirectmicroscopic
examination,byaddingadropofKOH(20%),putacover
slideonit,thenallowforabout15minutesthendiagnosedby
lightmicroscopedirectly
2. Cultural examination:
TransferthespecimentoaPetridishcontainssterilized
SDA (Sabouraud DextroseAgar+Antibiotic
(Chloramphenicol) + Antifungal-Antisaprophyte-
(Cycloheximide)andthenincubateat37
o
Cfor10-15days