WhatisMycology?

ViralInfectionsBlogArticles

INTRODUCTIONTOMYCOLOGY
Theterm"mycology"isderivedfromGreekword"mykes"meaningmushroom.Thereforemycologyisthestudy
offungi.
Theabilityoffungitoinvadeplantandanimaltissuewasobservedinearly19thcenturybutthefirst
documentedanimalinfectionbyanyfunguswasmadebyBassi,whoin1835studiedthemuscardinediseaseof
silkwormandprovedthethattheinfectionwascausedbyafungusBeauveriabassiana.
In1910RaymondSabouraudpublishedhisbookLesTeignes,whichwasacomprehensivestudyof
dermatophyticfungi.Heisalsoregardedasfatherofmedicalmycology.
Importanceoffungi:
Fungiinhabitalmosteverynicheintheenvironmentandhumansareexposedtotheseorganismsinvarious
fieldsoflife.
BeneficialEffectsofFungi:
1.Decompositionnutrientandcarbonrecycling.
2.Biosyntheticfactories.Thefermentationpropertyisusedfortheindustrialproductionofalcohols,fats,
citric,oxalicandgluconicacids.
3.Importantsourcesofantibiotics,suchasPenicillin.
4.Modelorganismsforbiochemicalandgeneticstudies.Eg:Neurosporacrassa
5.SaccharomycescerviciaeisextensivelyusedinrecombinantDNAtechnology,whichincludestheHepatitis
BVaccine.
6.Somefungiareedible(mushrooms).
7.Yeastsprovidenutritionalsupplementssuchasvitaminsandcofactors.
8.PenicilliumisusedtoflavourRoquefortandCamembertcheeses.
9.ErgotproducedbyClavicepspurpureacontainsmedicallyimportantalkaloidsthathelpininducinguterine

contractions,controllingbleedingandtreatingmigraine.
10.Fungi(LeptolegniacaudateandAphanomyceslaevis)areusedtotrapmosquitolarvaeinpaddyfields
andthushelpinmalariacontrol.
HarmfulEffectsofFungi:
1.Destructionoffood,lumber,paper,andcloth.
2.Animalandhumandiseases,includingallergies.
3.Toxinsproducedbypoisonousmushroomsandwithinfood(MycetismandMycotoxicosis).
4.Plantdiseases.
5.Spoilageofagricultureproducesuchasvegetablesandcerealsinthegodown.
6.Damagetheproductssuchasmagnetictapesanddisks,glasslenses,marblestatues,bonesandwax.
Generalpropertiesoffungi:
1.Theyareeukaryoticcellscontainmembraneboundcellorganellesincludingnuclei,mitochondria,golgi
apparatus,endoplasmicreticulum,lysosomesetc.Theyalsoexhibitmitosis.
2.Haveergosterolsintheirmembranesandpossesses80Sribosomes.
3.Havearigidcellwallandarethereforenonmotile,afeaturethatseparatesthemfromanimals.Allfungi
possesscellwallmadeofchitin.
4.Arechemoheterotrophs(requireorganiccompoundsforbothcarbonandenergysources)andfungilack
chlorophyllandarethereforenotautotrophic.
5.Fungiareosmiotrophictheyobtaintheirnutrientsbyabsorption.
6.Theyobtainnutrientsassaprophytes(liveoffofdecayingmatter)orasparasites(liveoffoflivingmatter).
7.Allfungirequirewaterandoxygenandtherearenoobligateanaerobes.
8.Typicallyreproduceasexuallyand/orsexuallybyproducingspores.
9.Theygroweitherreproductivelybybuddingornonreproductivelybyhyphaltipelongation.
10.Foodstorageisgenerallyintheformoflipidsandglycogen.
Classificationoffungi:
Fungiwereinitiallyclassifiedwithplantsandwereasubjectofinterestforbotanistshencetheinfluenceof
botanycanbeseenontheirclassification.In1969R.HWhittakerclassifiedalllivingorganismsintofive
kingdomsnamelyMonera,Protista,Fungi,PlantaeandAnimalia.

Traditionallytheclassificationproceedsinthisfashion:
Kingdom
Subkingdom
Phyla/phylum
Subphyla
Class
Order
Family

Genus
Species
Thisclassificationistoocomplicatedtobedealthere.
Therearealternateandmorepracticalapproaches,onebasedonsexualreproductionandtheother
basedonmorphologyofthethallus(vegetativestructure).

BasedonSexualreproduction:
1.Zygomycetes:whichproducethroughproductionofzygospores.
2.Ascomycetes:whichproduceendogenoussporescalledascosporesincellscalledasci.
3.Basidiomycetes:whichproduceexogenoussporescalledbasidiosporesincellscalledbasidia.
4.Deuteromycetes(Fungiimperfecti):fungithatarenotknowntoproduceanysexualspores(ascospores
orbasidiospores).Thisisaheterogeneousgroupoffungiwherenosexualreproductionhasyetbeen
demonstrated.
BasedonMorphology:
1.Moulds(Molds):FilamentousfungiEg:Aspergillussps,Trichophytonrubrum
2.Yeasts:SinglecelledcellsthatbudsEg:Cryptococcusneoformans,Saccharomycescerviciae
3.Yeastlike:SimilartoyeastsbutproducepseudohyphaeEg:Candidaalbicans
4.Dimorphic:Fungiexistingintwodifferentmorphologicalformsattwodifferentenvironmentalconditions.
Theyexistasyeastsintissueandinvitroat37oCandasmouldsintheirnaturalhabitatandinvitroatroom
temperature.Eg:Histoplasmacapsulatum,Blastomycesdermatidis,Paracoccidiodesbrasiliensis,
Coccidioidesimmitis.
Some200"humanpathogens"havebeenrecognizedfromamonganestimated1.5millionspeciesoffungi.
Morphologyoffungi:
Fungiexistintwofundamentalformsthefilamentous(hyphal)andsinglecelledbuddingforms(yeast).But,for
theclassificationsaketheyarestudiedasmoulds,yeasts,yeastlikeanddimorphicfungi.Allfungihavetypical
eukaryoticmorphology.Theyhaverigidcellwallcomposedofchitin,whichmaybelayered
withmannans,glucansandotherpolysaccharidesinassociationwithpolypeptides.Somelowerfungipossess
celluloseintheircellwall.SomefungisuchasCryptococcusandyeastformofHistoplasmacapsulatum
possesspolysaccharidecapsulesthathelpthemtoevadephagocytosis.Innertothecellwallistheplasma
membranethatisatypicalbilayeredmembraneinadditiontothepresenceofsterols.Fungalmembranes
possessergosterolincontrasttocholesterolfoundinmammaliancells.Thecytoplasmconsistsofvarious
organellessuchasmitochondria,golgiapparatus,ribosomes,endoplasmicreticulum,lysosomes,microtubules
andamembraneenclosednucleus.Auniquepropertyofnuclearmembraneisthatitpersiststhroughoutthe
metaphaseofmitosisunlikeinplantandanimalcellswhereitdissolvesandreforms.Thenucleuspossesses
pairedchromosomes.
Moulds:

Thethallusofmouldismadeofhyphae,whicharecylindricaltubelikestructuresthatelongatesbygrowthat
tips.Amassofhyphaeisknownasmycelium.Itisthehyphathatisresponsibleforthefilamentousnatureof
mould.Thehyphaemaybebranchedorunbranched.Theymaybeseptateoraseptate.Hyphaeusuallyhave
crosswallsthatdividethemintonumerouscells.Thesecrosswalls,calledseptahavesmallporesthrough
whichcytoplasmiscontinuousthroughoutthehyphae.Thereforeallhyphalfungitendtobecoenocytic
(multinucleate).Withexceptionofzygomycetes(Rhizopus,Mucor),allmouldsareseptate.Nonseptatehyphae
areconsideredtobemoreprimitivebecauseifahyphalstrandisdamagedtheentirestranddies.Whena
septatehyphalstrandisdamaged,theporesbetweenadjacentcompartmentscanbeplugged,thuspreventing
deathofthewholehyphalstrand.
Myceliumareofthreekinds:

1.Vegetativemyceliumarethosethatpenetratesthesurfaceofthemediumandabsorbsnutrients.
2.Aerialmyceliumarethosethatgrowabovetheagarsurface
3.Fertilemyceliumareaerialhyphaethatbearreproductivestructuressuchasconidiaorsporangia.
Sincehyphaisthestructuralunitofmould,themyceliumimpartscolour,textureandtopographytothe
colony.Thosefungithatpossessmelaninpigmentsintheircellwallarecalledphaeoidordematiaceousand
theircoloniesarecolouredgrey,blackorolive.ExamplesarespeciesofBipolaris,Cladosporium,
Exophiala,Fonsecaea,PhialophoraandWangiellaThosehyphaethatdon'tpossessanypigmentintheircell
wallarecalledhyaline.Hyphaemayhavesomespecialisedstructureorappearancethataidinidentification.
Someoftheseare:
a)Spiralhyphae:ThesearespirallycoiledhyphaecommonlyseeninTrichophytonmentagrophytes.
b)Pectinatebody:Theseareshort,unilateralprojectionsfromthehyphaethatresembleabrokencomb.
CommonlyseeninMicrosporumaudouinii.
c)Favicchandelier:Thesearethegroupofhyphaltipsthatcollectivelyresembleachandelierortheantlers
ofthedeer(antlerhyphae).TheyoccurinTrichophytonschoenleiniiandTrichophytonviolaceum.
d)Nodularorgan:Thisisanenlargementinthemyceliumthatconsistsofcloselytwistedhyphae.Oftenseen
inTrichophytonmentagrophytesandMicrosporumcanis.
e)Racquethyphae:Thereisregularenlargementofoneendofeachsegmentwiththeopposingend
remainingthin.SeeninEpidermophytonfloccosum,Trichophytonmentagrophytes.
f)Rhizoides:Thesearetherootlikestructuresseeninportionsofvegetativehyphaeinsomemembersof
zygomycetes.

g)Therearestructuresinthehyphae,whichariseoutofmodificationofasinglecellandtransformintothick
walledrestingcells.Chlamydospore(orchlamydoconidia),whichareproducedbyTrichophyton
schoenleiniiandTrichophytonverrucosumarethickwalledcellsthatarelargerthanothercellsandarranged
singlyoringroups.InsomefungisuchasTrichosporonbeigeilliandCoccidioidesimmitissomealternating
cellsbecomethickwalledandsubsequentlytheinterveningcellsdisintegrateleavingbehindarthrospores
(orarthroconidia).

Yeasts:
Yeastsareunicellularsphericaltoellipsoidcells.Theyreproducebybudding,whichresultinblastospore
(blastoconidia)formation.Insomecases,asthecellsbudsthebudsfailtodetachandelongatethusforming
achainofelongatedhyphaelikefilamentcalledpseudohyphae.ThispropertyisseeninCandiaalbicans.The
samespeciesalsohavetheabilitytoproducetruehypha,whichisseenasgermtube.Thedifferencebetween
thetwoisthatthereisaconstrictioninpsueudohyphaeatthepointofbudding,whilethegermtubehasno
constriction.

SomeyeastsuchasCryptococcusandtheyeastformofBlastomycesdermatatidisproduce
polysaccharidecapsule.CapsulescanbedemonstratedbynegativestainingmethodsusingIndiainkor
Nigrosin.ThecapsuleitselfcanbestainedbyMeyerMucicarminestain.Someyeastsarepigmented.
Rhodotorulaspsproducespinkcoloniesduetocarotenoidpigmentswhilesomeyeastssuchas
PhaeoannellomyceswerneckiiandPiedraiahortaearedematiaceous,producingbrowntoolivaceous
colonies.TrueyeastssuchasSaccharomycescerviciaedon'tproducepseudohyphae.Yeastlikefungimay
bebasidiomycetes,suchasCryptococcusneoformansorascomycetessuchasCandidaalbicans.
Reproductioninfungi:
Fungireproducebyasexual,sexualandparasexualmeans.
Asexualreproductionisthecommonestmodeinmostfungiwithfungiparticipatinginsexualmodeonly
undercertaincircumstances.Theformoffungusundergoingasexualreproductionisknownasanamorph(or
imperfectstage)andwhenthesamefungusisundergoingsexualreproduction,theformissaidtobe
teleomorph(orperfectstage).Thewholefungus,includingboththeformsisreferredasholomorph.
(Taxonomically,theteleomorphortheholomorphisused,butpracticallyitismoreconvenienttousethe
anamorph.)
Asexualreproduction:
Asexualpropagulesaretermedeithersporesorconidiadependingontheirmodeofproduction.Asexualspores
areproducedfollowingmitosiswhereassexualsporesareproducedfollowingmeiosis.
Theasexualsporesofzygomycetes,whichareknownassporangiosporesformwithinsaclikestructureknown
assporangia.Thesporangiosporesresultfromthemitoticcleavageofcytoplasminthesporangium.The
sporangiaareborneonspecialhyphaecalledsporangiophore.Thisendogenousprocessofsporeformation
withinasacisknownassporogenesis.

Conidiaariseeitherbybuddingoffconidiogenoushyphaeorbydifferentiationofpreformedhyphae.These
developfollowingmitosisofaparentnucleusandareformedinanymannerexceptinvolvingcytoplasmic
cleavage.Thisexogenousprocessisknownasconidiogenesis,aprocessthatoccursbothinyeastsand
moulds.Conidiaareborneonspecialisedstructurescalledconidiophore.Conidiaproductionmaybeblasticor
thallic.Inblasticdevelopmenttheconidiumbeginstoenlargeandaseptumisformed.Heretheconidium
originatesfrompartofparent.Inthallicmodeofdevelopmenttheconidiumisdifferentiatedbyaseptumbefore
itsdifferentiation.Thustheconidiumresultsfromtheconversionofentireparentcellintotheconidium.Thecell
thatgivesrisetoaconidiumiscalledaconidiogenouscell.Conidiophoresarespecialisedhyphaethat
bearconidiaorconidiogenouscells.Inmanycasesconidiogenouscellsarereferredasphialides.
SexualReproduction:

Sexualpropagulesareproducedbythefusionoftwonucleithatthengenerallyundergomeiosis.
Thefirststepinsexualmethodsofreproductioninvolvesplasmogamy(cytoplasmicfusionoftwocells).The
secondstepiskaryogamy(fusionoftwocompatiblenuclei),resultinginproductionofdiploidorzygotenucleus.
Thisisfollowedbygeneticrecombinationandmeiosis.Theresultingfourhaploidsporesaresaidtobesexual
spores,e.g.zygospores,ascosporesandbasidiospores.
Ifasexualsporeisproducedonlybyfusionofanucleusofonematingtypewithanucleusofanothermating
type(+andstrains),thefungusissaidtobeheterothallic.Incontrast,homothallicmouldsproducesexual
sporesfollowingthefusionoftwonucleifromthesamestrain.Forsexualreproductiontooccur,twocompatible
isolatesarerequired.Zygospores,whicharethesexualsporesofzygomycetesareround,thickwalled
reproductivestructuresthatresultfromtheunionoftwogamentagia.Ascomycetesproducesexualspores
calledascosporesinaspecialsaclikecellknownasascus.Inbasidiomycetesthebasidiosporesarereleased
frombasidium,whichistheterminalcellofahyphae.
Parasexualreproduction:

Parasexualreproduction,firstseeninAspergillusisknowntooccurinbasidiomycetes,ascomycetesand
deuteromycetes.Theprocessinvolvesgeneticrecombinationwithouttherequirementofspecificsexual
structures.

ImportanceofSpores:

A.Biological

1)Allowsfordissemination
2)Allowsforreproduction
3)Allowsthefungustomovetonewfoodsource.
4)Allowsfungustosurviveperiodsofadversity.
5)Meansofintroducingnewgeneticcombinationsintoapopulation
B.Practical
1)Rapididentification(alsohelpswithclassification)
2)Sourceofinoculaforhumaninfection
3)Sourceofinoculaforcontamination
ZYGOMYCETES
Commonlyknownasbreadmoulds,thesearefastgrowing,terrestrial,largelysaprophyticfungi.Hyphaeare
coenocyticandmostlyaseptate.Asexualsporesincludechlamydoconidia,conidiaandsporangiospores.
Sporangiophoresmaybesimpleorbranched.Sexualreproductioninvolvesproducingathickwalledsexual
restingsporecalledazygospore.
Medicallyimportantordersandgenerainclude:
1.Entomophthorales:ConidiobolusandBasidiobolusareinvolvedinsubcutaneouszygomycosis
2.Mucorales:Rhizopus,Mucor,Rhizomucor,AbsidiaandCunninghamellaareinvolvedinsubcutaneous
andsystemiczygomycosis(formerlycalledMucormycosis)
BASIDIOMYCETES
Theyexistassaprobesandparasitesofplants.Hyphaearedikaryoticandcanoftenbedistinguishedbythe
presenceofclampconnectionsoverthesepta.Sexualreproductionisbytheformationofexogenous
basidiospores,typicallyfour,onabasidium.Occasionalspeciesproduceconidiabutmostaresterile.
Generaofmedicalimportanceinclude:
1.TeleomorphofCryptococcusneoformans,whichisFilobasidiellaneoformans
2.AgentsofbasidiomycosissuchasCoprinusandSchizophyllium
3.MushroompoisoningbyAminita,Lepiota,CoprinusandPsilocybeetc.
ASCOMYCETES
Theyexistassaprophytesandparasitesofplants.Hyphaeareseptatewithsimpleseptalpores.Asexual
reproductionisbyconidia.Sexualreproductionisbytheformationofendogenousascospores,typicallyeight,in
anascus.
Medicallyimportantgeneraincludethe:
1.Teleomorphsofknownpathogenicfungie.g.Arthroderma(ofTrichophytonandMicrosporum),
Ajellomycesdermatitidis(ofBlastomycesdermatitidis),Pseudallescheriaboydii(ofScedosporium
apiospermum)
2.Agentsofmycetoma,likeLeptosphaeria

3.Agentsofblackpiedra,likePiedraiahortae.
DEUTEROMYCETES
DeuteromycetesarealsoknownasFungiImperfectibecauseofabsenceofsexuallyreproducingforms
(teleomorphorperfectstage).Astheirteleomorphcontinuetobediscovered,theywouldbeclassifiedamong
thepreviouscategories,untilthenthisremainsanartificialandheterogeneousgroup.Therearethreeclasses
ofFungiImperfecti.
1.Blastomycetes:TheseincludeasexualbuddingformsofCryptococcus,Candida,Torulopsisand
Rhodotorula.Dependingonthepresenceofmelaninintheircellwalls,theymaybenondematiaceousor
dematiaceous.
2.Hyphomycetes:Aclassofmycelialmouldswhichreproduceasexuallybyconidiaonhyphae.Hyphae
areseptate.Thisclasscontainsthemajorityofmedicallyimportantfungi.Dematiaceoushyphomycetesare
thoseconidialfungithatproducedarkbrown,greenblack,orblackcoloniesandarethecausativeagents
ofphaeohyphomycosis.Hyalinehyphomycetesincludethoseconidialfungi,whicharenotdarklypigmented
coloniesmaybecolourlessorbrightlycoloured.Theseincludetheagentsofhyalohyphomycosis,
aspergillosis,dermatophytosisandthedimorphicpathogens,likeHistoplasmacapsulatum.
3.Coelomycetes:Theseproduceacervuli,whicharetightlyboundmatsofhyphaeonwhichconidiaare
produced.
Pathogenesisoffungaldiseases(Mycoses):
Mostfungiaresaprophyticorparasitictoplantsandareadaptedtotheirnaturalenvironment.Infectionin
humansisachanceevent,occurringonlywhenconditionsarefavourable.Exceptforfewfungisuchasthe
dimorphicfungithatcausesystemicmycosesanddermatophytes,whichareprimarypathogens,therestare
onlyopportunisticpathogens.Humanbodyisahostileenvironmentandoffersgreatresistancetofungal
invasion.Mostfungiaresaprophyticandtheirenzymaticpathwaysfunctionmoreefficientlyattheredox
potentialofnonlivingsubstratesthanattherelativelymorereducedstateoflivingmetabolizingtissue.Some
fungisuchasCandidaandMalasezziahaveadaptedtohumanenvironmentandexistascommensals.
Thecomplexinterplaybetweenfungalvirulencefactorsandhostdefencefactorswilldetermineifafungal
infectionwillcauseadisease.Infectiondependsoninoculumsizeandthegeneralimmunityofthehost.
FungalPathogenicity(virulencefactors):
Abilitytoadheretohostcellsbywayofcellwallglycoproteins
Productioncapsulesallowingthemtoresistphagocytosis
ProductionofacytokinecalledGMCSFbyCandidaalbicansthatsuppresstheproductionofcomplement.
AbilitytoacquireironfromredbloodcellsasinCandidaalbicans
Abilitytodamagehostbysecretingenzymessuchaskeratinase,elastase,collagenase
Abilitytoresistkillingbyphagocytesasindimorphicfungi
Abilitytosecretemycotoxins
Havingauniqueenzymaticcapacity

Exhibitingthermaldimorphism
Abilitytoblockthecellmediatedimmunedefencesofthehost.
Surfacehydrophobicity
Hostdefencefactors:
Physicalbarriers,suchasskinandmucusmembranes
Thefattyacidcontentoftheskin
ThepHoftheskin,mucosalsurfacesandbodyfluids
Epithelialcellturnover
Normalflora
Chemicalbarriers,suchassecretions,serumfactors
Mostfungiaremesophilicandcannotgrowat37oC.
NaturalEffectorCells(polymorphonuclearleucocytes)andtheProfessionalPhagocytes(monocytesand
macrophages)
Factorspredisposingtofungalinfections:
Prolongedantibiotictherapy
Underlyingdisease(HIVinfection,cancer,diabetes,etc.)
Age
Surgicalprocedures
Immunosuppressivedrugs
Irradiationtherapy
Indwellingcatheters
Obesity
Drugaddiction
Transplants
Occupation
Immunitytofungalinfections:
Mechanismofimmunitytofungalinfectionscanbeinnateoracquired.Thenonspecificimmunityincludesthe
physicalbarriersofferedbyskinandmucusmembranesalongwiththeirsecretionsandnormalflora.ThepH,
bodytemperatureandserumfactorsalongwithphagocyticcellsplayanimportantpartinprovidingnon
specificimmunity.Eventhoughbodymountsbothhumoralandcellmediatedimmunity,itisthelatterthatisthe
mainstayofhostdefence.
Cellmediatedimmunity:
Immunityisprovidednonspecificallybeeffectorcells(polymorphonuclearleucocytes)andprofessional
phagocytes(monocytesandmacrophages)andspecificallybyTlymphocytes.Thephagocytesarevery
importantindefenceagainstCandia,AspergillusandZygomycetesasisevidencedbytheirseverityin
granulomatousdiseases,myeloperoxidasedeficiencyandcytotoxicchemotherapy.
ExpressionofTcellmediatedimmunitytofungiincludes:

delayedtypehypersensitivity
contactallergy
chronicgranulomatousreactions
Humoralimmunity:
Eventhoughantibodiesareproducedagainstmanyfungi,theirroleinprotectionisnotveryclear.However,
antibodieshelpinclearingfungalpathogensthroughopsonisation,whichisimportantagainstCandidaand
Cryptococcus.Anothercomponentofhumoralimmunityisthecomplement,whichcanactasopsoninsand
mayevencausedamagetotheircellsthroughcomplementactivation.Antibodiesareimportanttofungal
serodiagnosis.
Hypersensitivity:
Asaresultofdermatophyteinfectionsomefungusfreeskinlesionsofvariablemorphologyoccurelsewhereon
thebody,whicharethoughttoresultfromhypersensitivitytothefungus.Thesereactionsarecalled"id
reaction".ThesereactionsarealsoseennCandidainfections.Aninflamedboggylesionofthescalpcalledthe
kerionmayresultfromastrongimmunereactiontothedermatophyte.Granulomasduetointracellularfungi
representdelayedhypersensivities.Manyfungiaresignificantallergenstohumans,theallergensbeingspores,
conidia,hyphaeandotherfungalproducts.Oninhalationtheymayproduceallergicpulmonarydiseasessuch
asallergicbronchopulmonaryaspergillosis,farmer'slung,maplebarkstripper'slung,bronchialasthmaetc,
whichmaybeTypeIorIIIhypersensitivity.
FungalDiseases(Mycoses):
Mycosescanbeconvenientlystudiedas:
1.Superficialmycoses
I.Superficialphaeohyphomycosis
II.Tineaversicolor
III.Blackpiedra
IV.Whitepiedra
2.Cutaneousmycoses
I.Dermatophytosis
II.Dermatomycosis
3.Subcutaneousmycoses
I.Chromoblastomycosis
II.Rhinosporidiasis
III.Mycetoma
IV.Sporotrichosis
V.Subcutaneousphaeohyphomycosis
VI.Lobomycosis
4.Systemic(deep)mycoses

I.Blastomycosis
II.Histoplasmosis
III.Coccidioidomycosis
IV.Paracoccidioidomycosis
5.Opportunisticmycoses
I.Candidiasis
II.Cryptococcosis
III.Aspergillosis
6.Othermycoses
I.Otomycosis
II.Occulomycosis
7.Fungalallergies

8.Mycetismandmycotoxicosis

Laboratorydiagnosisofmycoses:
Specimencollection:Specimencollectiondependsonthesiteaffected.Differentspecimensincludehair,
skinscrapings,nailclippings,sputum,blood,CSF,urine,cornealscraping,dischargeorpusfrom
lesionsandbiopsy.

Allspecimensmustbetransportedtothelaboratorywithoutanydelaytopreventbacterialovergrowth.
Incaseofdelayspecimensexceptskinspecimen,bloodandCSFmayberefrigeratedforashortperiod.
Infectedhairsmaybepluckedusingforceps.ThosehairsthatfluoresceunderWoodslampmaybe
selectivelyplucked.Hairsmaybecollectedinsterilizedpaperenvelopes.
Surfaceoftheskinmustbedisinfectedwithspiritbeforespecimencollection.Theadvancingedgeofthe
lesionisscrapedwiththehelpofabluntforcepsandcollectedinsterilizedpaperenvelopes.
Discolouredorhyperkeratoticareasofnailmaybescrapedordiseasednailclippingmaybecollectedin
sterilizedpaperenvelopes.
Specimensfrommucusmembranes(oral)mustbecollectedbygentlescrapingandtransportedto
laboratoryinsteriletubecontainingsaline.Swabsmaybecollectedfromvagina.
Cornealscrapingsmaybecollectedusingafineneedleandinoculatedatbedside.

Pusmaybecollectedbyaspirationuseofcottonswabsmaygivefalsepositivemicroscopicresults.
Cleancatchurinemaybecollectedinasterilewidemouthedcontainer.
Biopsyspecimensmustbetransportedinsaline.
Incertaincases,pusorexudatesmustbelookedforpresenceofgranules.
Microscopy:
Microscopyisusedtoobserveclinicalspecimensforthepresenceoffungalelementsortoidentifythefungus
followingculture.Inthelattercase,lactophenolcottonblueisstainofchoice,whichstainsthefungalelements
blue.Directexaminationofclinicalspecimenscouldbestainedorunstained.
Wetmount:Candidamaybeobservedinurinewetmounts
1020%KOHmount:SeveralspecimensaresubjectedtoKOHmountfordirectexamination.Thematerialis
mixedwith20%KOHonaslideandacoverslipisplaced.Theslideisthengentlyheatedbypassingthrough
theflame23times.Theslideisobservedoncooling.KOHservestodigesttheproteindebrisandclears
keratinisedtissueandincreasesthevisibility.AdditionofDimethylsulphoxide(DMSO)permitsrapidclearingin
theabsenceofheat.
Calcofluorwhite:Thisisafluorescentdye,whichbindsselectivelytochitinofthefungalcellwall.The
specimenthencanbeobservedunderfluorescentmicroscope.
IndiaInk:CapsulesofCryptococcusneoformanscanbedemonstratedbythisnegativestainingtechnique.
PeriodicAcidSchiff(PAS)stain:Onstainingbythisstain,fungalelementsappearbrightmagenta
colouredwhilethebackgroundstainsgreen.Itisusefulinstainingtissuespecimens.
Giemsasstain:ItisparticularlyusefulinthedetectionofHistoplamsacapsulatuminthebonemarrow
smears.
HaematoxylinandEosin(H&E)stain:Usefulforstainingtissuesections.
Gomorismethenaminesilvernitrate(GMS)stain:Outlinesofthefungiareblack,internalpartsstain
pinkblackwhilethebackgroundstainslightgreen.CandidaandAspergillusmaybemissedinH&Estained
sections,thereforeGMSstainedsectionsareessentialfortissuepathology.
Gridleysstain:Itstainshyphaeandyeastsdarkbluepink,tissuesdeepblueandbackgroundyellow.
Meyermucicarminestain:CapsulesofC.neoformansandinnerwallsofRhinosporidiumseeberis

sporangiumarestainedpink.
Gramstain:CandidaisbestdemonstratedinclinicalspecimenbyGramstain.
MassonFontanastainishelpfulinstainingphaeoid(dematiaceous)fungiintissue.
Immunofluorescence:Monoclonalantibodylabelledwithfluorescentdyescanbeusedtodetectseveral
fungiintheclinicalspecimens.
Culture:OneofthemostcommonmediausedtoculturefungiinlaboratoryisSabouraudsDextroseAgar
(SDA).Itconsistsofpeptone,dextroseandagar.HighconcentrationofsugarandalowpH(4.55.5)prevents
growthofmostbacteriaandmakesitselectiveforfungi.EmmonsmodificationofSDAcontains2%dextrose
andhaspHof6.8.
OtherbasalmediatogrowfungiincludePotatoDextroseAgar,MaltExtractAgaretc.Mostfungiareableto
growatroomtemperaturewhilefewpathogenicfungi(e.g,Cryptococcus,dimorphicfungi)cangrowat37oC.
Saprophyticfungigrowmuchquicklythanpathogenicfungi(e.g,dermatophytes).Insuchsituationsthe
saprophyticfungicanbeinhibitedbytheadditionofcycloheximide(actidione)totheSDA.Additionofantibiotics
suchasChloramphenicol,GentamicinorStreptomycintoSDAservestoinhibitbacterialmultiplication.An
exampleofSDAwithcycloheximideandChloramphenicolisMycoselagar.
Otherspecializedmediausedfordifferentfungiinclude:
BrainHeartInfusionAgargeneralisolationoffungiandconversionofdimorphicfungi.
InhibitoryMouldAgar,anisolationmediumwithChloramphenicoltosuppressmostbacteria.
CaffeicAcidAgarandBirdseedAgarforisolationofCryptococcusneoformans.
CornMealAgar:EnhancesproductionofchlamydosporesinCandidaalbicansandformationofconidia
infungi.
TrichophytonAgars:UsedforselectiveidentificationofTrichophytonspecies.
DermatophyteTestMedium:Usedforrecoveryofdermatophytesfromclinicalspecimens.
SabhiMedium:IsolationofHistoplasmacapsulatum.
CHROMagarCandidaisusefulinidentificationofCandidaspecies.
Conversionofmouldtoyeastphasemustbedemonstratedinvitroforidentificationofdimorphicfungi.Since
somefungigrowslowlyculturesshouldnotbediscardedfor46weeks.Fungiareidentifiedonthebasisof
colonymorphology(includingpigmentation)andmicroscopicobservationbyteasemountpreparationorslide
culturetechnique.
Serology:Detectionofantifungalantibodyishelpfulindiagnosisofsubcutaneousandsystemicmycoses,
prognosisandresponsetoantifungaldrugs.Differentserologictechniquesthatareusedincludeagglutination,
immunodiffusion,counterimmunoelectrophoresis,complementfixationtest,immunofluorescence,RIAand
ELISA.
Antigendetection:ItisparticularlyusefulinthediagnosisofcryptococcalmeningitisfromCSFspecimens.
The

testisperformedbyLatexAgglutinationorimmunodiffusiontests.Itisalsohelpfulinthedetectionof
AspergillusandCandidaantigensinsystemicinfections.
Skintests:Delayedhypersensitivityreactionstofungalantigenscanbedemonstratedbyskintests.Apositive
skindoesnotnecessarilyindicateanactiveinfectionitonlyindicatessensitizationoftheindividual.Hence,its
valueisinepidemiologicalstudiesthandiagnosis.ThesetestsmaybeperformedinHistoplasmosis,
Candidiasis,Sporotrichosis,Coccidioidomycosis,Blastomycosis,Paracoccidiodomycosisanddermatophytosis.
Moleculartechniques:NewertechniquessuchasDNAhybridization,PCRareusefulindiagnosisofmycoses
inashorterperiodaswellasdetectthosefungithataredifficultordangeroustocultivateinvitro.