INTRODUCTIONTOPARASITOLOGY

INTRODUCTION
I.SYMBIOSIS(Sym=Together;Bio=LivingLivingTogether)
anassociationbetweentwodifferentspeciesoforganisms
primaryaim:toobtainfoodandseekshelterandprotectionfromthebiggerorganism
commonlyknownasLivingTogether
therearethreetypesofSymbioticRelationships:Commensalism+Mutualism+Parasitism
ORGANISM
Commensalism
Mutualism
Parasitism

Favorable
Favorable
VeryFavorable

HOST
NotAffected
Favorable
NotFavorable

II.PARASITOLOGY
thestudyoftheparasites
thesciencethatdealswithaspecializedgroupofanimalswhicharephysiologicallydependentonother
organismsfortheirsurvival
Neva&Brown:sciencethatdealswithorganismsthattakeuptheirabodes,temporarilyorpermanently,onor
withinotherlivingorganismsforfood,withtherelationshipoftheseorganismstotheirhost
**IMPORTANTNOTE:
o ThekeyinstudyingParasitologyisintheLifeCycle
o Ifweknowthelifecycleanditsimportantcomponents,wewouldunderstandwhyitisthekey
III.DEFINITIONOFTERMS
A.Parasites
organismsrangingfrommicroscopictomacroscopicwhichareknowntoinflictdiseaseinman
exhibitgreatdiversityinmorphologyandphysiology
animalsdependingonbiggerorganismsforfoodandshelter
hasintimatealmostalwaysobligatoryrelationshipwiththehost(ALWAYSharmsthehost)
**Ascarislumbricoides
competeforavailablenutrients(sugars,proteins,fats)inthefoodwehadalreadyeaten
tangledwormsmayobstructthegut
maydamagevitalorgansduringerraticmigration
**Description:
HarmsitsHost
Associatedcloselywithhostandderivenutritionfromhost
Maykillthehost
B.Hosts
thebiggerorganismsinwhichtheparasitelives
providesthenourishmentandprotectionneededbytheparasites
**Description:

Doesnotharmtheparasites
Providessheltertoitsparasite(parasitesneedthehost)

PARASITES
I.CLASSIFICATIONOFPARASITES
Parasites:organismoranimalthatdependsuponanotherbiggerorganismforfood
parasitescanbeclassifiedaccordingtolocationintheirhosts,degreeofassociation,andtaxonomicgroups
A.AccordingtoLocation
1.Ectoparasites
foundoutsideoronthesurfaceofthebodyofthehost
maycauseINFESTATION
ex)Scabies(Sarcoptesscabieihumanitchmite)
2.Endoparasites
foundlivingwithinthehostinanyinternalorgans(GIT,RespiratoryTract,RBCs)
maycauseINFECTION
examplesofEndoparasites:
Ascaris(GITLumen)
Plasmodium(RBC)MalarialParasite
Schistosoma(MesentericVeins)
B.AccordingtoDegreeofAssociation
1.ObligateParasite
thosewhocannotsurviveorreproduceoutsidethehost
alwaysparasitic(mustalwaysbeincontactwithasusceptibleorganism)
ex)Trichomonasvaginalis
2.FacultativeParasite
thosethatmayexistinafreelivingstateorasacommensal
ifopportunitypresentsitself,theymaybecomeParasitic
theycanbothliveasfreelivingorparasiticparasites
ex)Strongyloidesstercoralis
3.PermanentParasite
ifthelivecontinuouslyinthehost,andremaininthebodyofthehostfromtheearlystage
untilmaturity
requirescontinuouscontactwiththehost
ex)Pediculus,Trichomonasvaginalis
4.Intermittent(Temporary)Parasites
contactwiththehostisonlytemporarilycloselyassociatedwithitshosttemporarilytofeed
ex)Anophelesmosquitoes(theyonlybitewhentheywantyourblood)
5.AccidentalParasites
entersthehostthatisnotnormallyitshost
doesnotdeveloptofullmaturitysincethehostisnotphysiologicallycompatible
associationlastsonlyforashorttime
ex)Echinococcus,Gnathostoma,Toxocara
6.AberrantParasite
movesoutofthenormallocationwithinitshost
itsMigrationtoectopicsitesmayleadtocomplications
alsoknownaserratic

ex)Ascarislumbricoides(intestines,thentheymigratetobileduct,liver,orappendix)
7.SpuriousParasite
organismstakeninbyaccidentandthenpassedoutwithoutcausinganyharmtounsuitablehost
exampleofCommnesalism(exFruitFlylarvaeorPlantNematode)

II.TYPESOFHOST
Host:organisminwhichaparasiteslive
classifiedaccordingtothefollowing:

A.DefinitiveHost
harborsthesexualorthematurestageoftheparasite(sitewheresexualreproductionoccurs,ifpresent)
ex)Man,Pig,Dog,Mosquitoes
Mosquitoes:definitivehostinMalaria
Ascaris:manisthedefinitivehost
B.IntermediateHost
hostthatharborstheasexualortheimmature/larvalformsoftheparasites
sitewheretheasexualreproductionoccurs
ParatenicHost:hostinwhichtheparasitedoesnotundergoanydevelopment
C.ReservoirHost
thesearetheanimalsthatharbortheparasitethatarealsoparasitictoman
sitewheretheasexualreproductionoccurs
ex)Dogmaycarryhookwormeggsfromoneplacetoanother,buteggsdonothatchorpassthrough
anydevelopmentintheseanimals
D.Accidental(Incidental)Host
thesearethehoststhataccidentallyingestedtheParasites(thatusuallyaffecttheanimalkingdom)
infectionoccursinahostotherthanthenormalhostspecies
Zoonosis:conditionwhereinanimalparasitesaccidentallyinfectman
ex)HyatidTapewormsareusuallyfoundindogsandsheepZoonosis
E.Vector
aninvertebrateanimalthattransmitsaparasiticorganismfromonehosttoanother
hastwotypes:MechanicalVector+BiologicalHost
1.MechanicalVector

aninvertebrateanimalwhichmerelytransferparasiticorganismsfromonehost
toanother(bycarryingfecesontheirfeet)
ex)Flies,Cockroaches

2.BiologicalVector

thereisadevelopmentoftheparasiteinsidethehost
harbortheparasiteinternallyinwhichitmayundergofurtherdevelopment,
multiplication,orboth
ex)MosquitoesinMalaria

III.TAXONOMICGROUPINGSOFPARASITES
A.Protozoans(UnicellularEukaryoticParasites)
o PhylumSarcomastigophora
o PhylumApicomplexa
o PhylumMicrospora
o PhylumCiliophora
PHYLUM
REPRODUCTION
Sarcomastigophora:
Asexual
Sarcodina
Sarcomastigophora:
Asexual
Mastigophora
Apicomplexa

Asexual&Sexual

Ciliophora

LOCOMOTION
Pseudopodia

SPECIES
Entamoebahistolytica

Flagella

Giardialamblia
Dientamoebafragilis
Trypnasomaspp
Loishmaniaspp
Plasmodiumspp
Toxoplasma
Sarcocystis
Balantidiumcoli

None
Cilia

1.PhylumSarcomastigophora
hasSubphylum:SarcodinaandMastigophora
bothreproduceAsexually
a.SubphylumSarcodina

movebyPseudopodia(canonlymoveinonedirection)
Pseudopodia:cytoplasmicprotrusions
ex)Amoeba,Entamoeba(pathogen)

b.SubphylumMastigophora

movebyflagella(Flagellates)
Flagellum:longthreadlikeextensionofcytoplasm
ex)Giardia,Trypanosoma,Leishmania,Trichomonas

2.PhylumApicomplexa
nolocomatoryorgan
haveacomplexlifecyclewithalternatingsexualandasexualgenerations
BloodParasitesthatcauseMalaria:Isospora,Cyclospora,Cryptosporidium,Sarcocystis
OtherParasites:Taxoplasma,Sarcocystis
3.PhylumCiliophora
movebybeatingofmanyCilia
mostlyfreelivingandsymbioticspecies
ex)Balantidiumcoli(theonlypathogenicciliate)
4.PhylumMicrospora

nolocomatoryorgan
intracellularparasiteofmanykindsofvertebratesandinvertebrates
rarelycausediseaseinimmunocompetentpersons

B.Helminths(Metazoa;WormlikeInvertebrates)
PHYLUMASCHELMINTHES
CLASSNEMATODA
AKA
Roundworms
Shape
Elongated,Cylindrical
Segmentation
Unegmented
Sexes
Separate
1.PhylumAschelminthes
a.ClassNematoda

PHYLUMPLATYHELMINTHES
CLASSCESTODA
CLASSTREMATODA
Flatworms
Flukes
Elongated,RibbonLike
Leafshaped,Elongated
Segmented
Unsegmented
Hermaphrodite
Hermaphrodite

NematodesorRoundworms
elongated,cylindricalwormsattenuatedatbothends
unsegmentedworms;sexesareseparate
reproductionby:
Oviparous(ex.Ascaris,Enterobius)
Viviparous(ex.Trichinella,FilarialWorms)
**Infectionby:
IngestionofEggs(ex.Ascaris,Trichuris)
SkinpenetrationbyLarvae
VectorBorne
IngestionofEncystedLarvae

2.PhylumPlatyhelminthes
alsocalledFlatwormsmulticellularanimalscharacterizedbyflat,bilaterallysymmetricbody
mostFlatwormsareHermaphroditic,havingbothmaleandfemalereproductivesystems
a.ClassTrematoda

majorityaretransmittedbyingestion(exceptSchistosoma)
unsegmented
Hermaphroditic
flattened,leafshapedorcylindrical
OrderDigenea:containsallspeciesthatareparasitictohumans
Infectionby:MetacercariaeORSkinpenetrationbyCercariae

b.ClassCestoda

segmented
Hermaphroditic,ribbonlike
reproductionby:Oviparous(Ex.Taenia);multiplicationoflarvalforms
Infectionby:IngestionofEncystedLarvae

C.Arthropods
1.PhylumArthropoda
havesegmentedbodyparts:HeadThoraxandAbdomen
havethreetofourpairsofjointedappendagesandcoveredbyhardchitinuousskeleton
bilaterallysymmetricalbody
**ClassesinPhylumArthropoda

CLASS
Crustacea
Chilopoda
Arachnida
Insecta

DESCRIPTION
Someserveasintermediatehosts
spiderlike
Includesmostimportantofarthropods

EXAMPLE
Crabs,shrimps,copepods
Centipedes
Scorpions,spiders,ticks,mites

**ClassInsecta
OrderAnoplura
OrderHemiptera
OrderColeoptera
OrderHymenoptera
OrderSiphonaptera
OrderDiptera

LIFECYCLES
I.TWOTYPESOFLIFECYCLES:
ParasitesundergoaLifeCycleduringitsexistence
itinvolvessurvivalanddevelopmentintheexternalenvironmentandinoneormorehosts
lifecycleconsistsofthevariousmorphologicformsanddevelopmentalstages
lifecyclesummarizesdevelopmentoftheparasitefromthetimeitleavesthehosttostartanewinfectionina
newsusceptiblehost
knowledgeoflifecycles=preventivemeasures
A.DirectLifeCycle
thereisnointermediatehostrequiredtotransmittheparasitefromonehosttoanother
parasitesaretransmittedfromaninfectedindividualtoasusceptiblehostwithoutrequiringan
intermediatehost
**Ex1)Trichomonasvaginalis
directcyclewheretheparasiteisimmediatelyinfectivewhenitleavesthehost
ManMan
**Ex2)Ascarislumbricoides
directlifecyclewheretheparasitedevelopsintotheInfectiveStageintheexternal
environment
ManDevelopmentinExternalEnvironmentVectors(optional)Man
B.IndirectLifeCycle
atleastoneintermediatehost/vectorisrequiredfortheparasitetocompleteitstransmissionfromthe
originallyinfectedhosttothesusceptiblepotentialnewhost
theinfectivestageoftheparasitedevelopsintheIntermediateHostandtheSusceptibleHostacquires
theinfectionthroughcontactorexposuretotheinfectedintermediatehost
**Ex)Schistosomajaponicum,Capillariaphiippinensis
ManIntermediateHostContact/IngestionofRawIntermediateHostMan

II.COMPONENTSOFTHELIFECYCLE
InfectiveStage
PortalofEntry
ModeofTransmission
PathorRouteofMigration
DevelopmentalStages
Habitat
PortalofExit
DiagnosticStage
A.InfectiveStage
stageofdevelopmentoftheparasitewhichwheningestedtothedefinitivehostwillresulttoinfection
inthedirectlifecycle:InfectiveStagecomesfromoriginalinfectedhost
intheindirectlifecycle:itoriginatesfromtheIntermediateHostorVector
ex)AscarisInfectedStageistheEmbryonatedEgg
B.PortalofEntry
referstositeofopeninginthesusceptiblehostthroughwhichtheinfectivestageenters
mostcommonportalofentry:Mouth,Skin,Nose
ex)AscarisPortalofEntry:Mouth
C.ModeofTransmission
referstotheprocessonhowanewhostacquirestheinfectivestageoftheparasite
mancouldacquireitby:Ingestion,Inhalation,InfectedIntermediateHost,Skinpenetration,Inoculation
ex)inAscarisIngestionofEmbryonatedEgg;HookwormbySkinpenetrationofLarva
D.PathorRouteofMigration
whentheinfectivestageofparasiteentersthenewhost,itusuallyundergoeswithoutmaturationstages
differentdevelopmentalstagesofparasitesmayoccurindifferentpartsororgans
E.DevelopmentalStages
theparasiteundergoesdifferentdevelopmentalstagesasitmovesfromtheoriginalinfectedhoststo
theenvironment,insidetheintermediatehosts&evenw/inthenewsusceptiblehostbeforesettlingdown
ex)ForHelminths,developmentalstageisEgg,Larva,ImmatureAdultandAdult

F.Habitat
referstothespecificorganorsiteinthedefinitivehostsbodywherethesexuallymatureparasiteresides
ex)HabitatofAscarisisintheLumenofSmallIntestines
G.PortalofExit
openingorsiteinbodywhereinparasitesleavetheinfectedhoststofindanewsusceptiblehost
ex)EggsofAscarisleavethehostthroughtheAnus
H.DiagnosticStage
stagepassedoutbythehostortakenupbythevectororintermediatehost
servesasabasisforestablishingpresenceoftheparasiteinhostordiagnosisofinfection

EXPERIMENT24:LUNGFLUKES

INTRODUCTION
I.SLIDESTOVIEW:
Paragonimuswestermani
o Adult
o Egg
II.PARAGONIMUSWESTERMANI
Paragonimuswestermani=AffectstheHumanLungs(LungFluke)
Transmission=IngestionofInsufficientlycookedCrabsorCrayFishinfectedwithMetacercariae
InfectiveStage=Metacercaria
Diagnosis=FindingMediumSizedOperculatedOvaintheSputum
Pathology=Acute,SubacuteInflammationsofLungTissue;itmayalsowandererraticallytoBrainandSkin
**IMPORTANTNote:
o Clinically,itisusuallyMistakenwithTuberculosisSymptomsduetoHemoptysis
o TODifferentiate,wefindOperculatedOvaintheSputum,insteadofAcidFastBacilli
III.LIFECYCLEOFP.WESTERMANI
Type=IndirectLifeCycle(OneorTwoIntermediateHostsandOneDefinitiveHost)
ImmatureLarvaedevelopsinsidetheIntermediateHostandMaturesintoAdultStageinsideMan
ModeofTransmission=IngestionofRawInfectedCrabs/Crayfish
DiagnosticStage=EggsinSputum/Stool
InfectiveStage=MetacercariaefromCrabs
DefinitiveHost=Man
1stIntermediateHost=Snails
2ndIntermediateHost=Crabs/Crayfish

DEFINITIVEHOST=MAN
IngestionofRawInfected
Crabs/Crayfish
ImmatureWorm
MigrateintoPeritonealCavity,
DiaphragmandLungs

AdultWorm
EncystedinLungTissue
(LungshaveCavitaryLungLesions,
Cough,PleuriticPain,Hemoptysis)
Eggs
inSputumorStool

SNAILS

CRABS/CRAYFISH

Miracidium
Hatches&PenetratesSnail
Sporocyst

Cercariae
EntertheCrabs
EncyststoformMetacercaria

Radiae
Cercariae

Metacercariae
(InfectiveStage)

Cercariaeisreleasedinthe
Water

III.DESCRIPTIONOFTHESLIDES:
A.ParagonimuswestermaniAdult
Plump,Ovoid
ReddishBrownFluke(820mminlength;59mminbreadth)
**DiagnosticFeaturesofthisFluke
ScaleLikeIntegumentalSpines
TwoEqualSizedVentral&OralSuckerslocatedintheAnteriorHalf
**OtherStructures:
TwoLobateTestessituatedSidebySide
CentrallyLocatedLobateOvaryAnteriortotheTestes
Laterally,theentirebodyisoccupiedbytheHighlyBranchedVitellaria
ExcretoryBladder
CoiledUterus
B.ParagonimuswestermaniEgg
MediumSized,OvalShapedEgg(85x55u)
Operculated:hasacharacteristicBroaderOperculatedAnteriorEndandaThickerPosteriorEnd
expectoratedinSputumorwhenswallowed,itcanbedetectedinFecesinUnembryonatedStage

IV.ANSWERSTOQUESTIONS
DiscusstheSpecimensofChoiceinconfirmingtheDiagnosisofParagonmiasis
o Stool
o Sputum

 GivetheCharacteristicMorphologicalFeatureofParagonimuswestermaniOva
o BroaderOperculatedAnteriorEnd
o ThickenedPosteriorEnd

EXPERIMENT27:SPOROZOANS

INTRODUCTION

Sporozoans=GroupofUnicellularParasiteswithacharacteristicApicalComplex
HasNOLocomotoryOrganelles(exceptforFlagellatedMicrogametesinsomegroups)
Plasmodium=CommonlyseeninManwhichcancauseMalaria
ClassificationofSporozoans(BasedonAffectedOrgan)
o BloodSporozoans
o TissueSporozoans

I.SLIDESTOVIEW:
P.falciparum=Trophozoites,Schizonts,Gametocytes
P.vivax=Trophozoites,Schizonts,Gametocytes
P.malariae=Trophozoites,Schizonts,Gametocytes
P.ovale=Trophozoites,Schizonts,Gametocytes
II.PROCEDURE
StainedBloodSmearsareshown
PayattentiontotheSizeoftheNormalandParasitizedRBC,numberofChromatinDots,AmountofHemozoin
Pigments,AppearanceoftheCytoplasm,andpresence/absenceofStipplingontheInfectedRBCsCytoplasm
III.STRUCTUREOFTHEPARASITE(PLASMODIUM)

10

HostRBC
Stippling
Cytoplasm
Chromatin

Vacuole

IV.LIFECYCLEOFMALARIAPARASITES
TypeofLifeCycle=IndirectCycle
InfectiveStage=Sporozoites
DiagnosticStage=Gametocytes
BiologicVector=FemaleAnophelesMosquito
DefinitiveHost=FemaleAnophelesMosquito
IntermediateHost=Human
TwoPhases=HumanPhase+MosquitoPhase
V.OTHERIMPORTANTSPOROZOANS:
Cryptosporidium
Isospora
Blastocystis
Pneumocystiscarinii
Toxoplasmagondii
V.STAGESOFTHEPARASITE
AsexualStages=Trophozoites+Schizonts
ImmatureSexualStages=Gametocyte
NOTE:ManisjustanIntermediateHostinMalarialInfectionsbecausetheyharboronlytheImmatureFormsof
theParasite(theMatureformsarefoundintheMosquito)
A.Trophozoites(RingForm)
itistheearlieststage
theydevelopfromSporozoitesinthePreErythrocytic(HepaticSchizogony)Cycle;intheBloodCycle,
itdevelopsfromMerozoitestherefore,weseeThrophozoitesinBOTHCyclesofHumanPhases
withONEorTWOChromatinDots
ringlikestructureswithFewMalarialPigment
**ChromatinDots:
UsuallyonlyONE!
P.falciparum(exception)=MorethanOneinTrophozoiteStage(thinringlikecytoplasm)
**Cytoplasms:
CompactCytoplasm=P.ovaleandP.malariae
ThinDelicateCytoplasm=P.falciparum
AmoeboidCytoplasm=P.vivax

11

BandformCytoplasm=P.malariae

B.Schizonts
oncetheChromatinDotdividesbyBinaryFission,ParasiteisnowaSchizont
withTwoorMorechromatinDots(ChromatinDotsnowdeveloptoformMerozoites)
withplentyorincreasingamountofMalarialPigment(HemozoinPigment)inCytoplasm
canbedividedintoGrowing/ImmatureandMature(Cryptozoites)inthePreErythrocyticCycle
**Merozoites
P.falciparum=1232
P.vivax=1224
P.malariae=612
P.ovale=412
**TissueSchizonts:
PrimaryTissueSchizonts
SecondaryTissueSchizonts
C.Gametocytes
itisonlyanImmatureformoftheSexualFormsoftheParasite
BananaShapedinP.falciparumorspecialwithdefiniteChromatinmaterial
developsfromMerozoites

DECSRIPTIONOFTHESPECIES
I.MORPHOLOGY(FromLabManual)
Trophozoite

Schizont

P.falciparum
Size:SmallMedium
Number:Numerous
Shape:Ring&CommaForm
Chromatin:TwoDots
Cytoplasm:Regular
MatureForms:Compact
Pigment:CoarseGrains

P.vivax
Size:SmallLarge
Number:FewModerate
Shape:BrokenRing
Chromatin:Single
Cytoplasm:Irregular
MatureForms:Compact
Pigment:FineScattered

P.malariae
Size:Small
Number:Few
Shape:RingtoRound
Chromatin:Single
Cytoplasm:Regular
Pigment:Scattered

P.ovale
Size:Smallerthanvivax
Number:Few
Shape:RingtoRound
Chromatin:Single
Cytoplasm:Regular
Pigment:Course,Scatter

Associatew/RingForms
Size:Small,Compact
Number:Few

Size:Large
Number:FewModerate

Size:Small,Compact
Number:Few

Size:likemalariae
Number:Few

Pigment:LooseMass

Pigment:Concentrated
DaisyFlowerLike

Pigment:Concentrated

Mature:Round
Pigment:Scattered

Mature:Round
Chromatin:Single

Mature:Round
Chromatin:Single

MatureForm:1213Merozoites

Pigment:SingleDarkMass
Gametocyte

Mature:BananaShaped
Chromatin:Single

MatureForm:1224Merozoite

MatureForm:612Merozoite

Mature:412Merozoite

12

Pigment:Scattered

Pigment:Scattered
Peripherally

Pigment:Scattered

II.GENERALCHARACTERISTICS(FromNotes)
FORM
InfectedRBCSize

P.falciparum
NotEnlarged

P.vivax
Enlarged

P.malariae
NotEnlarged

P.ovale
Enlarged

Shape

RoundbutmaybeCrenated

Round

Round

Oval;Fimbriated

Stipplings

MaurersDots

ShuffnersDots

ZiemansorJamesDots

ShuffnersDots

Trophozoites

SmallRings
Multiplew/AcoleForms

Small&LargeRings
Amoeboid

SmallCompactRings
BandForms

Small;CompactRings

Schizont

Rare(SeeninSevereCases)

IrregularContours

Compact;Round

Compact

#ofMerozoites

1232

1224

612
FormsRegularRosette
LikeClusters

412
FormsIrregularRosette
LikeClusters

Gametocyte

Banana/SausageShape

RoundLarge

Round,Compact

Round,Small

ANSWERSTOQUESTIONS

PlasmodiumfalciparumSchizontVSPlasmodiumvivaxSchizont
FEATURES
P.falciparum
Size
Small&Compact
Number
Few,Uncommon
MatureForms
1232Merozoites
Cluster
CompactClusters
Pigment
SingleDarkMass

P.vivax
Large
FewtoModerate
1224orMoreMerozoites
IrregularClusters
LooseMass

EXPERIMENT28:BLOOD&TISSUEFLAGELLATES

INTRODUCTION

TrypanosomaandLeishmaniaFlagellatesbelongtoOrderKinetoplastidaofSuperclassMastigophora
TheyMultiplyintheBlood(Hemoflagellates)orTissueofHumans
AllSpeciesrequireanArthropodIntermediateHost

I.SLIDESTOVIEW
Trypanosomabruceigambiense(Trypomastigote)
Leishmaniatropica(inCulture)
Leishmaniatropica(Hamster)
II.MORPHOLOGICDIFFERENCES
AKA
Morphology

AMASTIGOTE
Leishmania
Ovoidal/Spherical

PROMASTIGOTE
Leptomonas
SpindleShaped

EPIMASTIGOTE
Crithidia
SpindleShaped

TRYPOMASTIGOTE
Trypanosoma
SpindleShaped

13

Flagellum
Courseof
Flagellum
Foundin

IntracellularAxoneme
Intracellular(NotFree)

Stage

LeishmaniaDiagnosticStage

Macrophages
Reticuloendothelials

FreeFlagellum
ArisefromKinetoplast
intheAnteriorEnd
Vector

FreeFlagellum
EndsinKinetoplast
AnteriortoNucleus

FreeFlagellum
EndsinKinetoplast
PosteriortotheNucleus

LeishmaniaInfectiveStage

Typanosoma

TrypanosomaInfectiveStage

Leishmania=Amastigote+Promastigote
Trypanosoma=Trypomastigote+Epimastigote

III.TRYPANOSOMASPECIES
A.ComparisonofDiseasesandVectors
SPECIES
DISEASE
T.bruceigambiense
WestAfricanSleepingSickness
T.bruceirhodesiense
EastSleepingSickness
T.cruzi
ChagasDisease,Megacolon

VECTOR
RiverineTsetseFly
WoodlandTsetseFly
ReduviidBug(Triatoma)

B.LifeCycles:
1.T.bruceigambiense
InfectiveStage=MetacyclicTrypopmastigote
DiagnosticStage=Trypomastigote
StagethatdevelopsintheSalivaryGlandsoftheVector(TsetseFly)=Epimastigote
MammalianHosts=Humans,Pigs,WildAnimals
Vector=RiverineTsetsefly(Glossinapalpalis)=Daytimebiters
NOTE:EpimastigotesareNOTfoundintheHumansBlood(itdevelopsintheTsetseFly)
2.T.bruceirhodesiense
InfectiveStage=MetacyclicTrypopmastigote
DiagnosticStage=Trypomastigote
StagethatdevelopsintheSalivaryGlandsoftheVector(TsetseFly)=Epimastigote
NOTE:EpimastigotesareNOTfoundintheHumansBlood(itdevelopsintheTsetseFly)
3.T.cruzi
InfectiveStage=MetacyclicTrypomastigote
EpimastigoteDevelopsinsidetheReduviidBug(Vector)
DiagnosticPhases:
Blood=Trypomastigote
HeartTissues=Amastigote
IV.LEISHMANIASPECIES
A.ComparisonoftheDiseasesCaused:
LEISHMANIA
Leishmaniatropica

DESCRIPTION
OldWorldCutaneous
Leishmaniasis

DISEASES
Dry,Chronic,UrbanCutaneousLeishmaniasis
OrientalSore
AleppoorBaghadBoil
RecividansorChronicRelapsingCutaneousL.

Leishmaniamajor

OldWorldCutaneous
Leishmaniasis
OldWorldCutaneous
Leishmaniasis
NewWorldCutaneous

WetorAcuteRuralCutaneousLeishmaniasis

Leishmaniaaethiopica
Leishmaniamexicana

Diffuse/DisseminatedCutaneousLeishmanisis
Diffuse/DisseminatedCutaneousLeishmaniasis

14

Leishmaniasis

ChicleroUlcers

Leishmaniabraziliensis

Mucocutaneous
Leishmaniasis

MucocutaneousLeishmaniasis
Espundia(disfiguresface)

Leishmaniadonovani

VisceralLeishmaniasis

VisceralLeishmaniasis
KalaAzarorBlackDisease
(Zoonotic)

B.LifeCycle
o InfectiveStage=MetacyclicPromastigote
o DiagnosticStage=Amastigote(insideMacrophages)
o Vector=Sandflies(PhlebotomusorLutzomyia)
o ReservoirHosts=Humans,Dogs,WildAnimals
o Habitat=ReticularEndothelialSystem

EXPERIMENT29:FILARIALWORMS

INTRODUCTION

FilarialWorms=SlenderTissuedwellingNematodeswithReducedLipsandBuccalCapsules
AllSpeciesemployArthropodsasIntermediateHost
Transmission=throughArthropodVectorBite
InfectiveStage=ThirdStageLarva(L3)

15

I.SLIDESTOVIEW(BloodSmearswith):
Wuchereriabancrofti
Brugiamalayi
II.PROCEDURE
BloodSmearswithMicrofilariae
TakenoteofMorphologicalDifferencesbetweentheTwoSpeciesandTabulatebasedonthefollowing:Shape&
SizeoftheLarvaerelativetotheHostCells,presence/absenceofaMembraneSheathandNumberofNucleiat
thePosteriororTailEnd
III.FILARIALWORMSOFMEDICALIMPORTANCE
FILARIASIS
VECTOR
Wuchureriabancrofti
Lymphatic
Mosquitoes
Brugiamalayi
Lymphatic
Mosquitoes
Brugiatimori
Lymphatic
Mosquitoes
Onchoceriavolvulus
Subcutaneous
Fly
Loaloa
Subcutaneous
Fly
Manzonellaperstan
Animal
Midget
Manzonellaozzardi
Animal
Midget

DEFINITIVEHOST
Man
Man
Man
Man
Man
Animal
Animal

DISTRIBUTION
Philippines
Philippines
Indonesia

IV.LIFECYCLE
InfectiveStage=L3(ThirdStageLarvae)
DiagnosticStage=Microfilariae
DefinitiveHost=Man,Chimpanzee,Gorilla
MosquitoBorne
L3(InfectiveStage)MigratestoLymphGlandL1L2L3
HumanPhase

Mosquito

EXAMINATIONOFTHESLIDES
I.STAGESOFTHEPARASITE
A.AdultStage
foundinLymphGlands(theycancreateObstruction,leadingtoEdema)

16

longslender,creamywhiteroundworm,smoothcuticle

1.ViviparousFemale
VulvaopenstotheSurfaceoftheWormnearthemiddleofthePharynx
givesbirthtoaPrelarvalForm(Microfilariae)
610cminlength;0.3cmwide
2.Males
possessCaudalAlaewithtwoSpicules
4050mminlength,0.3wide
B.Microfilariae
alsoknownasthePreLarvalForm(L1)
itisNOTanInfectiveStageitisaDiagnosticStage
givenbirthbyaViviparousFemaleWorm(thereisNOeggstage)
BodyNuclei=representsareRudimentsofInternalOrgans
**Morphology:
MorphologyvarieswithSpecies
Maycontainsheathwithorwithoutstriations
BodyNucleirepresentRudimentofInternalOrgans
StainedwithGiemsa
**ComparisonofMicrofilariaebetweentheTwoSpecies:
W.bancrofti
CuticularSheath
Present
CephalicSpace
Short
Length=Width
BodyCurves
Regular,Large
BodyNuclei
RoundMediumSize
WellSeparated
Tail
NoTerminalNuclei
PointedEnd

B.malayi
Present
Long
Length>Width
Irregular,Kinky,Small
Small,Angular
Overlapping(notseparated)
WithTwoTerminalNuclei
BluntTip

*NOTE:MicrofilariaearefoundintheBloodonlyatNightTime
II.COMPARISONOFLYMPHATICANDSUBCUTANEOUSWORMS(BASEDONMICROFILARIAE)
W.bancrofti
Bmalayi
Onchocercavolvulus
Loaloa
Sheath
Present
Present
Absent
Absent
CephalicSpace
Short
Long
Short
Short
BodyCurves
Regular
Irregular
Rigid
Irregular
BodyNuclei
Separated
Overlapping
Separated
Overlapping
Tail
NoNuclei
WithTerminalNucleus
NoNucleus
WithTerminalNuclei
III.ENDEMICPLACESOFFILARIASISINTHEPHILIPPINES
Luzon
Leyte
Marinduque
Mindoro
Palawan
Samar

17

EXPERIMENT30:SCHISTOSOMA(BLOODFLUKES)

INTRODUCTION

Schistosoma=ComprisestheBloodFlukesortheBloodTrematodes
TheyonlyrequireONEIntermediateHosthaveNOMetacercarialStage
AdultshaveSeparateSexes

I.SLIDESTOVIEW:
Schistosomajaponicum=Egg,Cercaria,Adult
Schistosomamansoni=Egg,Adult
Schistosomamekongi=Egg,Adult
II.THREEMAJORSPECIES
SNAIL
MAJOR
INTERMEDIATEHOST
SPECIES
Schistosoma
OncomelaniaHupensis
japonicum
Quadrasi
Schistosoma
mansoni
Schistosoma
haematobium

Biomphalariaspp
(Biomphalariaglabrata)
Bulinusspp
(Bulinus18truncates)

HABITAT
VeinsoftheSmall
Intestines
VeinsoftheLarge
Intestines
Veinsof
GenitourinaryTract

DISEASE

DISTRIBUTION

OrientalSchistosomiasis
KatayamaDisease
Schistosomiasisjaponica
IntestinalBilharziasis
Schistosomiasismansoni
SchistosomalHematuria
VesicalSchistosomiasis
UrinaryBilharziasis

Philippines(MisamisOriental)
China,SulawesiinIndonesia,
Japan,Taiwan
Africa,SouthAmerica,West
Indies,PuertoRico
MiddleEast,Africa,Southern
TipofEurope

**NOTE:Schistosomajaponicum:producestheMostNumberoftheEggs
III.LIFECYCLE
InfectiveStage=Cercaria
DiagnosticStage=EggsinFeces
FormwhichEntertheSnails=Miracidia
IntermediateHost=Snails(differentspeciesforspecificspeciesofparasitesseetable)
ModeofTransmission=SkinPenetration

18

EXAMINATIONOFTHESLIDES
I.PROCEDURE:
NoteMorphologicFeaturesoftheParasitesandtheStrikingDifferencesbetweenthem
II.MORPHOLOGY
A.AdultWorms
0.62.5cminlengthwithOralandVentralSuckers
NarrowElongatedShape;hasseparateSexes;IntegumentisSmoothorTuberculateddependinguponspecies
numberofTestesinMalesandthelengthoftheUterus&NumberofEggsareDistinctivetothespecies
ExcretorySystem=FlameCells,CollectingTubulesleadingintoaSmallBladderwithTerminalExcretoryPore
MALEADULTWORM
ShorterbymoreSturdy;Grayish
HasaCylindricalAnteriorEnd
BodyFoldedtoformaGynecophoralCanal

FEMALEADULTWORM
Longer
MoreSlender
Darker

**GynecophoralCanal=FoundinMalesinwhichtheFemaleisembracedduringcopulation
B.Ova/Egg
withSpineorKnobLikeProjections
dependingontheStageofDevelopmentatthetimetheyarepassedoutwiththeFeces,onemayfind
themfromMulticellularStagetofullEmbryonationwithadevelopedMiracidiumwithintheShell
S.japonicumEgg
S.mansoniEgg
S.haematobiumEgg
*DevelopedMiracidium
*DevelopedMiracidium
*DevelopedMiracidium
*LateralKnob
*LargeLateralSpine
*LargeTerminalSpine
*SmallLateralSpine
C.Miracidium
ciliated,pyriforminshapewithaPrimitiveGut
D.Cercariae
forkedtailedwhenDischargedfromtheSnail
forksituatedatthePosteriorThirdoftheTail
OralandVentralSuckersaredeveloped
III.ANSWERSTOQUESTIONS
DifferentiatetheAppearanceoftheOvaoftheDifferentSpeciesofSchistosoma
S.japonicum
S.mansoni
S.haematobium
Oval
LightYellow=Brown
TerminalSpine
WithminuteSpineor Elongate
Knob
LateralSpine
AcidFast

S.mekongi
Oval
LateralSpineorKnob

WhatSpecimenareyougoingtocollecttodemonstratetheOvaofSchistotomajaponicum?
o FecesareCollected

19

WheneggscannotbefoundinFeces,aRectalBiopsymaybedone

EXPERIMENT36:INTESTINALAMOEBA

INTRODUCTION

ProtozoansintheIntestinesobtainnutrientsfromtheParasitizedHostorPredigestedFoodintheGIT
Trophozoite=ActiveorFeedingStage
Cysts=formwhentheEnvironmentisUnfavorableforSurvival(NotActive,NonFeeding)

I.SLIDESTOVIEW:
Entamoebahistolytica=Cyst,Trophozoite
Entamoebacoli=Cyst,Trophozoite
Entamoebanana=Cyst,Trophozoite
II.ENTAMOEBAHISTOLYTICA
TheONLYPathogen(othersareCommensals)
IthasthePotentialtoInvadetheIntestinalWall&otherOrgans
Entamoebadispar=anotherIntestinalCommensalwhichismorphologicallysimilarw/E.histolytica
TheycouldbedifferentiatedusingSpecificTests(IsoenzymeAnalysis,Immunoassays,PCR)
**NOTE:LIFECYCLEDiscussedBelow

EXAMINATIONOFTHESLIDES
I.TROPHOZOITESANDCYSTS:
TROPHOZOITES
*Karyosome=Center
*Nucleus=Eccentricw/FineChromatinGranules
*DirectionalMovements(ActiveProgressive)
*DefinedEctoplasm
*BladeLikeHyalinePseudopodia
*IndistinctNucleus
*IngestedRBC

CYSTS
*14Nuclei(MatureCyst)
*DiffuseGlycogenMass
*CigarShapedChromatoidalBars

Entamoebacoli

*Karyosome=EccentricKaryosomeinNucleus
*Nucleus=w/CourseChromatinGranules
*MoreSluggishNonDirectionalMovements
*MoreGranularCytoplasm
*ContainIngestedBacteria
*Narrower;LessDifferentiatedEctoplasm
*BlunterandBroaderPseudopodia
*Heavier
*IrregularPeripheralChromatin
*NORBCinside

*Asmanyas8Nuclei(MatureCyst)
*Larger;Slender
*MoreGranularCytoplasm
*SplinterLikeChromatoidalBodies

Endolimaxnana

*Karyosome=Eccentric,LargeIrregular

*Has4Nuclei

Entamoebahistolytica

20

*NoPeripheralChromatinGranules
II.ANSWERSTOQUESTIONS
WhatistheMorphologicFeatureofEntamoebahistolyticathatsignifiesPathogenicity?
o TrophozoiteofE.histolyticaisActive,MotileFeedingStagethatcausesPathologyintheColon
o ItisAnaerobicwithnomitochondria
o Erythrophagocytosis

EXPERIMENT37:INTESTINALFLAGELLATES&CILIATE

INTRODUCTION
I.SLIDESTOVIEW:
Giardialamblia
Dientamoebafragilis
Chilomastixmesnili
Balantidiumcoli
II.FLAGELLATESVSCILIATES
A.Flagellates
o PossessesoneormoreFlagellaLongThreadLikeExtrusionsoftheCytoplasm
o ThePathogenicIntestinalFlagellateofManisGiardialamblia
o ClassifiedintoTwoGroups(AccordingtoHabitat):
Intestinal,Oral,andGenitalFlagellates
BloodandTissueFlagellates
**NOTE:ALLhaveTrophozoiteandCysticStage(EXCEPTTrichomonas)
DientamoebafragiliswasoriginallygroupedunderAmoebas(nowitisaFlagellate)
Now,itisintheOrderTrichomonadida
DientamoebafragilishasTwoNucleiandNOCysticStage
B.Ciliates
o MovebymeansofCiliaThreadsofCytoplasm(shorterandmorenumerousthanFlagella)
o TheonlyPathogenicCiliateofManisBalantidiumcoli
III.REVIEWOFPROTOZOANS
Disease
InfectiveStage
Habitat
Diagnosis
Pathology
Transmission
DrugsofChoice
Trophozoite
Nucleus

Entamoebahistolytica
Amoebiasis
AmoebicDysentery
AmoebicHepatitis
MatureCyst
LargeIntestine(Cecum)
ParasiteinFeces
FlaskShapedUlcer

Trophozoite
LargeIntestines(Cecum)
BinucleateTrophozoites

Balantidiumcoli
Balantidiasis
Balantidosis
BalantidialDysentery
MatureCyst
LargeIntestines
Cysts&Trophozoites

Ingestion
Metronidazole

Giardialamblia
Giardiasis
Lambiasis
TravellersDiarrhea
MatureCyst
SmallIntestines
Cyst&Trophozoites
MalabsorptionSyndrome
SteatorrheacStool
Ingestion
Metronidazole

Dientamoebafragilis

Uncertain
Iodoquinol

Ingestion
Tetracycline

Single&Eccentric

TwoNuclei(Binucleate)

TwoNuclei(Binucleate)

Macro/Micro

21

Karyosome
Movement

Center
Unidirectional

Others

EasilyDestroyed

Center
4Flagella
TwistingMovements
SuckingDiskVentrally
PearShaped(OldMan)
2ParabasalBodies
2Axonemes

CigarChromatoidBodies
14Nuclei

Ellipsoidal
24Nuclei

Cyst
Description

Pseudopods
NonDirectional
NoFlagella

NONE

SurroundedbyCili
Cytosome&Cytopyge

ThickCystWall
RoundorOval

EXAMINATIONOFTHESLIDES
I.TROPHOZOITESANDCYSTS
Giardialamblia

TROPHOZOITE
*SuckingDiskonVentralSurface
*PearShaped
*OldMansFace
*TwoNucleiwithLargeCentralKaryosomes
*TwoAxonemes
*TwoBlepharoplasts
*TwoDeeplyStainingBars(ParabasalBodies)
*4PairsofFlagella

CYST
*24Nuclei
*SurroundedbyaCystWall
*Ellipsoidal(912um)

Dientamoebafragilis

*SluggishNonDirectionalMotility
*TwoNuclei
*NOFlagella!!!
*MayIngestRBC

NONE

Chilomastixmesnili

*OneNucleus
*FourAnteriorFlagella
*CytosomalGroove
*CurvedPosteriorly

*OneNucleus
*LemonPearShaped
*ClearKnob
*Cytosome

Balantidiumcoli

*SurroundedbyCili
*HasVacuolesandInclusionsintheCytoplasm
*Someuniquestructures:
Cytostome=PrimitiveMouth
Cytopyge=ExcretoryPore
Macronucleus=Bean/KidneyShaped
Micronucleus=withintheMacronucleus

*Almostsimilar:RoundorOval
*ThickCystWall

II.ANSWERSTOQUESTIONS
GivetheCharacteristicMorphologicFeatures

22

FLAGELLATE/CILIATE
Giardialamblia

LOCOMOTORYORGANELLE
14Flagella

DISTINCTFEATURES
TwoNuclei
LargeCentralKaryosome

Dientamoebafragilis

Amoebaflagellate

TwoNuclei(Binucleate)
NoCysts!

Chilomastixmesnii

Flagella

Trophozoite=TearDropShape
EvidentNucleus
HasaCystosome(CellMouth)
Cysts=LemonShaped

Balantidiumcoli

Cilia

Trophozoites:Large&Oval

EXPERIMENT38:INTESTINALNEMATODES

INTRODUCTION
I.SLIDESTOVIEW
A.AdultForms:
o Ascarislumbricoides
o Trichuristrichiura
o Enterobiusvermicularis
o Hookworms(Necatoramericanus,Ancylostomaceylanicum,Ancylostomacaninum)
o Capillariaphilippinensis
o Trichinellaspiralis
B.Larva
o Strongyloidesstercoralis
o Hookworms(Necatoramericanus,Ancylostomaceylanicum)
o Trichinellaspiralis
C.Ova
o Ascarislumbricoides(Fertilized,Unfertilized,andDecorticatedOva)
o Trichuristrichiura
o Hookworms(Necatoramericanus,Ancylostomaceylanicum)
o Capillariaphilippinensis
o Enterobiusvermicularis
II.NEMATODES
Nematodes/Roundworms=compriseaLargeGroupofHelminthsofsimplestructure
Elongated,Unsegmented,BilaterallySymmetrical
SeparateSexes
Males<Females
MaleshaveoneortwoCopulatorySpicules

23

EXAMINATIONOFTHESLIDES
I.OVERALLCHARACTERISTICS
Ascaris
Trichuris
lumbricoi
trichiur
des
a
GeneralCharacteristics
Common
GiantRound
Whipworm
Name
Worm
Diseases
Ascariasis
Trichuriasis
AscarisInfection
Trichocephaliasis

Enterobius
vermic
ularis
Pinworm/
Seatworm
Enterobiasis
Oxyuriasis

Hookworms

Threadworm

Ancylostomiasis
Uncinariasis
Necatoriasis

Strongyloides
Strongyloidosis
CochinChina

Capillariasis
PudocsDisease

Habitat

SmallIntestines
HeartLungRoute
FilariformLarva

SmallIntestines
HeartLungRoute
FilariformLarva

SmallIntestines

EggsinFeces

Hookworms

Strongyloides
stercoralis

Capillaria
philippinensis

SmallIntestines
HeartLungRoute
EmbryonatedEgg

LargeIntestines

LargeIntestines

EmbryonatedEgg

EmbryonatedEgg

EggsinStool

EggsinStool

EggsinFeces

Transmission
Definitive
Host
Intermediate
Host
Pathology

Ingestion
Man

Ingestion
Man(Monkeys)

EggsinPerianal
Region
Ingested
ManONLY

SkinPenetration
Man

Rhabditiform
Larva
SkinPenetration
Man

NONE

NONE

NONE

NONE

NONE

FreshwaterFish

Loefflers
Syndrome

RectalProlapse

PruritusAni

CreepingEruptions
MHAnemia
PneumoniaLike

LarvaCurens

Borborygmi
GurglingStomach

Treatment

Mebendazole
PyrantelPamoate
Albendazole

Mebendazole
Albendazole

Mebendazole
PyrantelPamoate
Albendazole

Mebendazole
PyrantelPamoate
Albendazole

Albendazole
Thiabendazole

Mebendazole
Albendazol
Thiabendazole

PointedEnds

BluntPosterior

SharpPointed
End

Parasitic
FreeLiving

CurvedPoserior
End

CurvedPosterior
SingleSpicule

CurvedPosterior
SingleSpicule

Atypical=23
Rows
Typical=1Row
LongSpicular
Sheath

Infective
Stage
Diagnosis

Morphology
FemaleAdult
MaleAdult

Bursatohold
female

NoParasiticMale
FreeLiving

ThirdStageLarva

Ingestion
Man

24

RetractileSheet
Eggs

Rhabditiform
Larva
Filariform
Larva
Distinct
Features

Fertilized=
Embryo
Unfertilized=
Granules

3BuccalLips

Lemon/Barrel
Shaped
BipolarMucus
Plug

AnteriorWhiplike
PosteriorRound

DShaped
WithEmbryo

CephalicAlae

28CellStage

LongBuccal
Capsule
PointedTail

PeanutShaped

ShortBuccal
Capsule
ForkedTail

RoundedEnds

II.RHABDITIFORM&FILARIFORMLARVAE
HOOKWORMS
RhabditiformLarva
BuccalCavity
GenitalPrimordium
FilariformLarva
Tail
Esophagus

LongBuccalCavity
SmallGenitalPrimordium

STRONGYLOIDES

ShortBuccalCavity
LargeGenitalPrimodrium

PointedTail
Notched/ForkedTail
ShorterEsophagus
LongerEsophagus
EXPERIMENT39:DIRECTFECALSMEAR

INTRODUCTION
I.PRINCIPLES
DirectFecalSmear(DFS)=SimplestandmostRapidofallFecalExaminationTechniques
RecommendedforIdentificationofProtozoanTrophozoitesandDetectionofHelminthicInfection
OneDirectFecalSmearPreparationcontainsapproximately2mgofFeces
Salineand/orLugolsIodineSolutioncanbeusedtoEmulsifytheFecalMaterial
A.UnstainedFilm
usefulfortheStudyofLivingParasiteObjects
ex)MotileProtozoanTrophozoites,HelminthEggs,andNematodeLarva
B.IodineFilm
employedtostudytheDiagnosticFeaturesofProtozoanCysts
II.PROCEDURE
Place12DropsSalineattheCenterofaGlassSlide
WithanApplicatorStick,pokeatvariousportionsoftheSpecimen(itwithbloodstreaks,makesureyoutouch
yourapplicatoratthisportion)
MakeasmoothuniformemulsioninthedropofsalinebyRotatoryMotionstartingfromtheCenter
PlacetheedgeofCoverSlipintheslidesothatittouchestheedgeofthewater
SlowlylowerthecoversliptopreventAirBubbles
Examineslide(firstwithLPO,thenwithHighPower)

25

ANSWERSTOQUESTIONS

GivetheAdvantageofusingLogolsIodineoverSalineSolutioninDirectFecalSmear
o LugolsIodineorDAntonisIodinehelpsvisualizeinternalstructuresofTrophozoitesandCysts
o BecauseStainswillkillMotileTrophozoites,itisrecommendedthataSalineSmearandanIodineSmear
bepreparedSeparately
BrieflyDiscusstheAdvantagesandDisadvantagesofaDirectFecalSmear
o Advantages
AllowstheviewertoDetectMotileProtozoa
CanviewHelminthEggs/Larvae,ProtozoanCysts,WBCs,SomeYeast
o

Disadvantages
ShouldNOTbeperformedonPreservedSpecimens
ShouldbeusedforFreshStoolSpecimensthatareverySoftorLiquid
CannotbeexaminedusingOilImmersion

EXPERIMENT40:CELLULOSETAPEPERIANALSWAB

INTRODUCTION
I.PRINCIPLES
PinwormInfection(Enterobiusvermicularis)issuspectedinchildrenwithPerianalItching,Insomnia,and
Restlessness
EvidencedependsonrecoveryofAdultWorms,Eggs,orBoth
TheyareRARELYFoundinStoolExamination
PinwormInfectioncanbebestdiagnosedbySwabbingPerianalAreausingGrahamsCellophane(Cellulose)
TapeMethod
II.GRAHAMSCELLOPHANETAPEMETHOD
HighestSensitivityandSpecificityResults
BesttoCollectSpecimensintheMorningbeforethePatientbathesordefecates
III.PROCEDURE
PlaceaStripofCelluloseTape,StickySideDown,onaslideandononeendaSmallStripofPaper
HoldaSlideAgainstaTongueDepressoroneinchfromtheendandliftthetapeawayfromtheSlide
LooptheTapeovertheendoftheTongueDepressortoexposetheGummedSurface(HoldtheTapebythePaper
TabattachedtoitsEndPortion
HoldtheTapeandSlideagainsttheTongueDepressor

26

PressthegummedsurfaceofthetapeagainstseveralareasofthePerianalArea
ReplacethetapeontheslideandexaminedirectlyunderthemicroscopeforPinwormEggsorTaeniaEggs

ANSWERSTOQUESTIONS

WhyisEarlyMorningtheBESTTimetocollectSpecimens?
o ThehighestPositiveResultsandGreatestNumberofEggscanbedetectedinthemorning
o SpecimensarebestobtainedafewhoursafterthePersonhasretired(1011pm)orthefirstthinginthe
morningbeforebowelmovementorbath

EXPERIMENT41:KATOTHICKSMEARANDKATOKATZTHICKSMEAR

KATOTHICKSMEAR(CELLOPHANETHICKSMEAR)
I.PRINCIPLES
KatoThickSmear=QualitativeMethodtodetectHelminthicInfections(betterthanDirectFecalSmear)
Requires5060mgFreshFecalMaterialsothatevenLightInfectioncanbedetected
CellophaneStripsusedisSoakedinGlycerineMalachiteGreenSolutionfor24hoursbeforeuse
ThismethodisveryusefulinMassExaminationofCommonSoilTransmittedHelminthicInfecitons
**GlycerineMalachiteGreenSolution
o Glycerine=clearstheFecalFilmtovisualizetheHelminthEggs
o MalachiteGreen=DyeusedtoprotecttheeyesfromIntenseLightNeededinexaminingThickSmear
o Preparation:500mlDistilledWater+500mlGlycerine+5ml3%MalachiteGreenSolutioninH 2O
**NOTE:ThisMethodisNOTSuitableforDiarrheicStoolandCANTDetectProtozoanCystsandTrophozoites
II.PROCEDURE
Place5060mgofStoolattheCenterofaGlassSlideandCoverwithaSquarePieceofPreTreatedCellophane

27

UsingaRubberStopper,presstheCellophanegentlytospreadtheStoolSpecimenevenly,approximatingthe
CircumferenceoftheRubberStopper
Leavethepreparedslideatroomtemperaturefor1020minutesduringthistime,theMicroscopicFieldbecomes
clearduetotheactionofGlycerineontheStoolConstituents
ExaminetheSlide(Theslideshouldbeexaminedafter1020minutesorwithin1hourafterpreparation
AllowingtheslidetostandforLongPeriodsoftimewillcauseDryingandShellsofHookwormOvawillbecome
transparentanddifficulttosee

III.ADVANTAGESVSDISADVANTAGES:
A.Advantage
o TimeSaving,Simple,Economical
o UsefulinMassExaminationofCommonSoilTransmittedHelminthInfections
B.Disadvantages:
o NotSuitableforDiarrheicStool
o CannotbeusedtodetectProtozoanCysts&Trophozoites

KATOKATZTHICKSMEAR(MODIFIEDTHICKSMEAR)
I.PRINCIPLES
KatoKatzThickSmearisaModificationofKatoThickSmearMethod
ItisaQuantitativeMethodforcountingHelminthicEggs
TheNumberofEggspergram(NPEG)ofFecescanbecomputedbyMultiplyingtheNumberofEggs
ObservedperThickSmearby24
II.PROCEDURE
PlaceaboutHalfaGramofFecalSampleonFilterPaper
Placethewirenetorscreenontopofthefecalsample
UsingaFlatSidedApplicatorStick,scrapeacrosstheuppersurfaceofthescreentosievetheFecalMaterial
PlaceaTemplateonaCleanMicroscopeSlide
TransferasmallamountofSievedFecalMaterialintotheholeoftheTemplateandcarefullyfillthehole
RemovetheTemplatecarefullysothatalltheFecalMaterialisleftontheslide

28

CovertheFecalSampleontheslidewithaGlycerineMalachiteSoakedCellophaneStrip
InverttheSlideandPresstheFecalSampleagainsttheCellophanegentlyonaSmoothSurfacetospreadevenly
Letstandforabout20minutesandexamine
Countalleggsseeninthewholepreparation
MultiplytheTotalEggscountedbyFactor24toexpresstheCountasEggsperGramFeces(EPG)
**IMPORTANTNotes:
o TheWireMeshNetisusedtoSeparateFecalMaterialfromtheLargeDebriswhichmayaffectthe
ApproximateFecalWeightandClearingofFecalFilmbyGlycerol

ANSWERSTOQUESTIONS

DiscusstheAdvantagesofThickSmearovertheDirectFecalSmear
o ThickSmearcanbeusedtoObtainanAccuratemeasureoftheNumberofEggsinagivenamountof
SamplesothattheWormBurdencanbeinferred
o However,AdultSchistosomestakeweekstomonthstopasseggsintotheFeces
GivetheRationaleofusingGlycerineMalachiteGreenSolutioninthesetwoprocedures
o ItisadyeusedtoDyetheSmearandProtecttheEyesfromtheIntenseLightthatisrequiredforthe
MicroscopicExaminationoftheSpecimen

EXERCISE42:INTESTINALTAPEWORMSANDFLUKES

INTRODUCTION

Cestoda(Tapeworms)&Trematoda(Flukes)ofPhylumPlatyhelminthes(orFlatworms)areExclusivelyParasitic

I.SLIDESTOVIEW
A.Trematodes
1.Adult,Egg
Fasciolopsisbuski
Echinostomailocanum
Clonorchissinensis

29

Opisthorchissp
Fasciolahepatica/gigantica

2.Adult
Fasciolahepatica
Metagonimusyokogaeai
Haplorchistaichui
3.Metacercaria
B.Cestodes
1.Adult
Taeniasolium
Taeniasaginata
2.Egg,Scolex,Mature/GravidProglottidof:
Taeniasolium
Taeniasaginata
Diphyllobothriumlatum
Hymenolepisnana
Hymenolepisdiminuta
Dipylidiumcaninum
3.Cysticercuscellulosae
II.TREMATODES(Flukes)
HaveConspicuousSuckers
LeafShaped,Unsegmented,andDorsoventrallyFlattened
ExternalCuticlewhichmostspeciesmaybecoveredwithScales,Spines,Tubercles,Ridges
TheyhaveComplexLifeCycles,involvingoneormoreIntermediateHosts
Transmission(Majority)=EatingInadequatelyCookedSecondIntermediateHostcontainingtheInfective
Metacercariae
III.CESTODES(Tapeworms)
EndoparasiticWormswithElongated,Segmented,andFlatBodydevoidofaBodyCavity
AdultTapeworm=Scolex(Head)+Neck+ProglottidsthatcomprisetheStrobilaorBodyoftheTapeworm
EachproglottidcontainsoneortwosetsofMaleandFemaleReproductiveOrgans

EXAMINATIONOFSLIDES:
I.TREMATODES(FLUKES)
A.IntestinalFlukes(Trematodes)
Disease
CommonName

Fasciolopsisbuski
Fasciolopsiasis
GiantIntestinalFluke

Echinostomailocanum
Echinostomiasis
GarrisonsFluke

Heterophyesheterophyes
Heterophyiasis
None

Metagonimusyokogawai
Mentagonimiasis
None

30

LifeCycle
DefinitiveHost

Indirect
Humans,Hogs,Dogs

1stIntermediate
2ndIntermediate
Habitat
InfectiveStage
DiagnosticStage
Morphology
Testes
Suckers
Egg
UniqueChar.

PlanorbidSnails
WaterPlants
SmallIntestines
Metacercaria
EggsinFeces

Indirect
Humans,Mammals,
Birds
PlanorbidSnails
Snails(Pilaluzonica)
SmallIntestines
Metacercaria
EggsinFeces

Indirect
Humans,FishEating
Mammals
BrakishWaterSnails
FreshwaterFish
SmallIntestines
Metacercaria
EggsinFeces

Indirect
Humans,FishEating
Mammals
Snails
FreshwaterFish
SmallIntestines
Metacercaria
EggsinFeces

Dendritic
Tandem
Oral+Ventral
Unembryonated
Operculated

Lobed
Tandem
Oral+Ventral
Unembryonated
Operculated

Lobate
SidebySide(Opposite)
Oral+Ventral+Genital
Embryonated
Operculated

Oval
ObliquelySidebySide
Oral+Ventral+Genital
Embryonated
Operculated

VitellariaLateralto
theCeca

SurroundedbyCrownof
Spines

FanShapedVitellaria

B.LiverFlukes(Trematodes)
Fasciolaspp

Clonorchissinensis

GeneralCharacteristics
Disease
Fascioliasis
LiverRot
PharyngealFascioliasis
Halzoun

Opisthorchisspp

Clonorchiasis

Opisthorchiasis

CommonName

SheepLiverFluke(F.hepatica)
LiverFluke(F.gigantica)

Chinese/OrientalLiverFluke

CatLiverFluke

LifeCycle
InfectiveStage
DefinitiveHost
DiagnosticStage
1stIntermediate
2ndIntermediate
ReservoirHost
Habitat

Indirect
Metacercariae
Man,Sheep,Cow,Deer,etc
OvainFeces
LymneidSnail
WaterPlants
BileDucts&BiliaryPassages

Indirect
Metaceracariae
ManandDog
OvainFeces
Planorbid(Operculate)Snails
FreshwaterFish(Cyprinoid)
None
BileDucts&BiliaryPassages

Indirect
Metaceracariae
ManandDog
OvainFeces
PlanorbidSnails
FreshwaterFish(Cyprinoid)
None
BileDucts&BiliaryPassages

LeafShape
Oral<Ventral
Dendritic;Tandem
FanShaped
Unembryonated
Branched
Branched
CephalicCone/Shoulder

Oblong,LanceolateShaped
Oral>Ventral
Branched;Tandem
Oval
Embryonated
Simple
Diffuse,IrregularlyDistributed
EggssmallerthanC.sinensis

Oblong,LanceolateShaped
Oral=Ventral
Lobate;Oblique
Oval
Embryonated
Simple
Cluster/CompressedFollicles

Morphology
Shape
Suckers
Testes
Ovary
Egg
IntestinalCeca
Vitellaria
UniqueFeature

II.CESTODES
Taeniasolium

Taeniasaginata

Diphyllobothrium
latum

Hymenolepisnana

Hymenolepis
diminu
ta

Dipylidium
caninum

31

GeneralCharacteristics
AKA
PorkTapeworm
Disease
Taeniasis
(Adult)

BeefTapeworm
Taeniasis

Diphyllobothriasis
(Adult)

DwarfTapeworm
Hymenolepiasis

RatTapeworm
Hymenoleipasis

DogTapeworm
Dipylidiasis

Sparganosis
(Larva)

Cysticercosis
(Larva)

Habitat
LarvalForm

BroadFishTapeworm

SmallIntestines
Cysticercus
cellulosae
Cysticercus
Egg

SmallIntestines
Cysticercusbovis

SmallIntestines
PlerocercoidLarva

SmallIntestines
Cysticercoid

SmallIntestines
Cysticercoid

SmallIntestines
Cysticercoid

Cysticercus

EmbryonatedEgg

Cysticercoid

Cysticercoid

GravidProglottids
EmbryonatedEgg

GravidProglottids
EmbryonatedEgg

PlerocercoidLarva
ProcercoidLarva
UnembryonatedEgg

EmbryonatedEgg

EmbryonatedEgg

ManOnly

ManOnly

Man,Dog,Cat,etc

Man,Mice,Rats

Rat,Mouse,Man

Embryonated
(Packets)
Dogs,Cats,Man

Pig

Cattle

Fleas,Louse

Ingestion

NONERequired
(RatFleas)
Ingestion

Fleas,Beetle

Ingestion

First:Copepod
Second:Fish
Ingestion

Ingestion

Ingestion

Globular
ArmedRostellum
FourSuckers

Pyriform
NoRostellum
FourSuckers

Spatulate/Almond
Bothria(Grooves)
Grooves

Globular
Retractile.Rostellum
FourSuckers

RowsofSpine
FourSuckers

Mature
Segment

TrilobedOvary

BilobedOvary
MoreTestes

BilobedOvary
Broader>Long
RosetteLikeUterus

BilobedOvary
3Testes

ClubShaped
Unarmed/
Rudiment
Rostellum
BilobedOvary
3Testes

Gravid
Segment
Egg

<13Branches

>13Branches

Embryonated

Embryonated

Embryonated

SaccularUterus
EggMasses
Embryonated

Infective
Stage
Diagnostic
Stage
Definitive
Host
Intermediate
Host
Transmission
Morphology
Scolex

Distinct
Feature

Unembryonated
LongestTapeworm

EggBiggerthan
H.nana

PumpkinSeed
Like
2SetsofRepro.
Organ
Membranous
Capsules
Embryonated
(Packets)
RiceGrain
Appearance

ANSWERSTOQUESTIONS
32

I.CLASSIFICATIONOFFLUKES
A.ClassificationofFlukesAccordingtoSize(ThoseinBoldLettersaretheIntestinalFlukes)
1.LargeFlukes(28cm)
Fasciolopsisbuski(largest)
Fasciolaspp
2.MediumFluke(12cm)
Echinostomailocanum
Clonorchissinensis
Opisthorchisspp
Paragonimuswestermanii
3.SmallFlukes(<1cm)
Heterophyesheterophyes
Metagonimusyokogawai
Phanerophsolusbonnie
Haplorchisspp

B.ClassificationofFlukesAccordingtoSecondIntermediateHost
1.WaterPlantBorne
Fasciolopsisbuski
Fasciolaspp.
2.SnailBorne
Echinostomailocanum
3.FishBorne
Heterophyesheterophyes
Metagonimusyokogawai
Clonorchissinensis
Opisthorchisspp

C.ClassificationofTrematodeEggs
1.LargeUnembryonatedEggs(100160u)
Fasciolopsisbuski
Echinostomailocanum
Fasciolaspp
2.MediumUnembryonatedEggs(7090u)
Paragonimuswestermanii
3.SmallEmbryonatedEggs(2332u)
Heterophyesheterophyes
Metagonimusyokogawai
Clonorchisspp
Opisthorchisspp

II.SCOLICESOFTHEDIFFERENTSPECIES:
CESTODE
TYPEOFSCOLEX
Taeniasolium
Globular
Taeniasaginata

Pyriform

SCOLEX
SuckingDisks(4)
RostellumwithChitinousHooks
SuckingDisks(4)

33

Diphyllobothriumlatum

Elongated/Spatulate

2ElongatedSuctorialGrooves=Bothria

MICROBIOLOGYPRACTICALEXAM:PARASITOLOGY
INTRODUCTION:
INTRODUCTIONTOPARASITOLOGY
EXPERIMENT24:
LUNGFLUKES
EXPERIMENT27:
SPOROZOANS
EXPERIMENT28:
BLOOD&TISSUEFLAGELLATES
EXPERIMENT29:
FILARIALWORMS
EXPERIMENT30:
SCHISTOSOMA(BLOODFLUKES)
EXPERIMENT36:
INTESTINALAMOEBA
EXPERIMENT37:
INTESTINALFLAGELLATES&CILIATE
EXPERIMENT38:
INTESTINALNEMATODES
EXPERIMENT39:
DIRECTFECALSMEAR
EXPERIMENT40:
CELLULOSETAPEPERIANALSWAB
EXPERIMENT41:
KATOTHICKSMEARANDKATOKATZTHICKSMEAR

34

EXERCISE42:
INTESTINALTAPEWORMSANDFLUKES

35