PARASITOLOGY( Intro & Protozoan) Flashcards
Branch of medicine or biology concerned with the phenomenon of dependence of one living organism with another living organism
Parasitology
Deals primarily with clinically significant parasites which are capable of causing a disease or infection to the host
Medical Parasitology
Study of protozoans (unicellular organism)
Protozoology
Unicellular organisms
Protozoans
Study of helminths (multicellular organisms)
Helminthology
Multicellular organisms
Helminths
Study of insects and arthropods that are of medical importance
Medical Entomology
Living together of phylogenetically different organims
Symbiosis
One organism gains benefits while the other organism neither benefits nor is harmed
Commensalism
Neutral to the other
Commensalism
Beneficial to one organism, harmful to the other
Parasitism
One organism benefits at the expense of the other
Parasitism
4 Types of Hosts
Definitive/ Final Host
Intermediate host
Paratenic Host
Reservoir host
Harbors the adult or sexual stage of parasite
Definitive
Harbors the larval or asexual stage of parasite
Intermediate
Allows the parasite’s life cycle to continue and serves as a source of human infection
Reservoir
Depends entirely upon its host for existence; cannot live outise of a host
Obligate
Capable of parasitic and free-living existence
Facultative
Capable of living independently outside of a host
Facultative
Larval stage is passed in a host while the adult is free living
Transitory
2 Types of Parasite according to duration of parasitism
Permanent
Temporary
Comple its life cycle in 1 host
Permanent
Parasites wherein all of its life cycle will happen in different host
Temporary
Deals with tropical diseases and other medical problems of tropical regions
Tropical Medicine
3 Types of Host-Parasite Relationship / Types of Association of Living Things
Mutualism
Commensalism
Parasitism
Beneficial to both organisms
Mutualism
Harbors a parasite that does not develop but remains alive and is infective to the next host
Paratenic
3 Example of Definitve host
Cat- DH of Toxoplasma gondii
Mosquito- DH of Plasmodium
Dog- DH of Echinococcus spp
1 Example of Intermediate host
Snail- IH of Trematodes/flukes
2 Example of Paratenic host
Large fish- PH of Diphylobothrium latum
Snake and Bird - PH of G. spinogerum
1 Example of Reservoir host
Pigs- RH of Balantidium coli
8 Types of Parasites according to mode of living
Endoparasite
Ectoparasite
Intermittent
Obligate
Facultative
Accidental
Erratic
Zoonotic
Parasites wherein all of its life cycle will happen only within a single host
Permanent
Requires 2 or more host for the larval and adult stages
Periodic
Primarily infects animals and may be acquired by man
Zoonotic
Occurs in an unsual host
Accidental
Occurs in an unusual organ or habitat
Erratic
Passes through the intestinal tract without causing any disturbance
Spurious or coprozoic
Lives inside the body of a host
Endoparasite
Lives on body surfaces
Ectoparasite
Visits the host only during feeding time
Intermittent
2 Example of Ectoparasite
Lice (Pediculus humanus capitis, P.h. humanus, Phthirus pubis)
Itch mite (Sarcoptes scabiei)
5 Example of Intermittent parasite
Bed bug
Mosquito
Flea
Tick
Biting fly
(BBM FT)
Presence of endoparasite
Infection
Presence of an ectoparasite
Infestation
5 Types of parasite accrording to pathologic locations
Spurious
Hematozoic
Cytozoic
Coelozoic
Enterozoic
Parasites that pass the gastrointestinal tract without infecting or causing harm to the host
Spurious
Parasites found in blood
Hematozoic
Parasites found in cells
Cytozoic
Parasites found in body cavities
Coelozoic
Parasites found within gastrointestinal tract
Enterozoic
2 Types of Vectors
Biologic
Mechanical/ Phoretic
Transmits a parasite only after the latter has completed part of its development
Biologic
Not essential in the parasite’s life cycle and is responsible only for transmitting the parasite
Mechanical/ Phoretic
3 Parasites transmitted by mosquitoes
Plasmodium spp.
Wuchereria bancrofti
Brugia malayi
3 Parasites transmitted by biting flies
Leishmania spp - Sandflies
Trypanosoma brucei - Tsetse flies
Other filariae -Loaloa
Parasites transmitted by kissing bugs
Trypanosoma cruzi
Parasites transmitted by ticks
Babesia spp.
Cockroaches and House flies is a mechanical vector of what parasites?
- Ascaris lumbricoides and
other enteric parasites (such as E. histoloytica)
MOT of mouth/oral cavity
Ingestion
Oral-anal intercourse
MOT of skin
Larval skin penetration
Skin inoculation (vector-borne)
MOT of urogenital tract
Sexual intercourse (venereal)
MOT of nasal passages
Intranasal
Type of Autoinfection within the intestine
Internal
MOT of Transplancental
Vertical transmission (mother to fetus)
Type of Autoinfection hand to mouth
External
2 Types of autoinfection
External
Internal
5 Example of portal of entry
Mouth/oral-anal cavity
Urogenital tract
Nasal passages
Transplancental
Skin
(MUNTS)
4 example of Portal of exit
Anus
Mouth (P. westermani)
Urogenital tract (T. vaginalis)
Non-intact skin (D. medinensis)
Study of patterns, distribution and occurence of disease
Epidemiology
Number of new cases of infection
Incidence
Number of individuals in a population estimated to be infected with particular parasite
Prevalence
Number of worms per infected person
Intensity of infection/ worm burden
Other term for intensity of infection
Worm burden
Use of anti-helminthic/parasitic drug in a public health program
Deworming
Permanent reduction to zero of the incidence of infection worldwide
Eradication
Reduction to zero of the incidence of infection in a specific region or geographical area
Elimination
Female cell within uterus
Ova
Early developing stage
Embryo
Motile, feeding stage of protozan
Trophozoite
Female cell outside uterus
Egg
Active stage of parasites
Larva
Non-motile, non-feeding stage of protozoan
Cyst
2 parasitic periodicity
Diurnal
Nocturnal
Day activity of parasites
Diurnal
Night activity of parasites
Nocturnal
3 Parasitic laying/giving birth characteristic
Viviparous
Ovo-viviparous
Oviparous
Parasite that do not lay eggs but instead produce fully developed larvae
Viviparous
Parasite that produce fully developed embryonated eggs
Ovo-viviparous
Parasite that produce unembryonated eggs that undergo external embryonation
Oviparous
4 Malarial terminologies
Microgametocytes
Macrogametocyte
Sporogony
Schizogony
Male sex cell of malarial parasites
Microgametocytes
Female sex cell of malarial parasite
Macrogametocyte
Sexual reproduction of malarial parasite
Sporogony
Sporogony takes place in?
Mosquito
Asexual reproduction of malarial parasite
Schizogony
Schizogony takes place in?
Man
2 Parasitic development
Encystation
Excystation
Trophozoite into cyst
Encystation
Cyst emerges into trophozoite
Excystation
3 Disease distribution
Endemic
Epidemic
Pandemic
Disease which is constant in a community
Endemic
Outbreak of a disease affecting a wide area
Epidemic
Worlwide epidemic
Pandemic
Represents the taxon to which the species belongs
Genus
First letter is capitalized
Genus
Trivial name that distinguishes the species within the genus
Specific epithet
3 Classification of Protozoa
Phylum Sarcomastigophora
Phylum Ciliophora
Phylum Apicomplexa
Both parts of the organism’s name are ____ in normal text or ______ in handwriting
Italicized
and
Underline
2 Classification of Sarcomastigophora
Amebae
Flagellates
1 Classification of Ciliophora
Ciliates - Balantidium coli
2 Classification of Phylum Apicomplexa
Sporozoans
Coccidians
2 Classification of Helminths
Phylum Nemathelminthes
Phylum Platyhelminthes
1 Type of Nemathelminthes
Nematodes
Other term of nematodes
Roundworm
2 types Platyhelminthes
Trematodes
Cestodes
Other term of trematodes
Flukes
Other term of cestodes
Tapeworms
3 Other unicellular parasites
Blastocytis spp.
Microsporidians
Pnemocystis jirovecii
3 Class of arthropods
Class Arachnida
Class Insecta
Class Crustacea
3 Example of Arachnida
Ticks
Mites
Chiggers
8 Example of Insecta
Lice
Fleas
Cockroaches
Bugs
Beetles
Flies
Mosquito
Midges
3 Example of crustacea
Crabs
Crayfish
Copepods
6 Example of stool preservatives
- 10% Formalin
- Schaudinn’s solution
- Polyvinyl alcohol (PVA)
- Merthiolate-iodine formalin (MIF)
- Sodium acetate-acetic acid formalin
- Modified PVA
All-purpose fixative
Formalin
5% concentration for protozoan cysts
Formalin
How many concentration of formalin for protozoan cysts
5%
5% formalin is used for
Protozoan cysts
10% concentration for helminth eggs and larvae
Formalin
How many concentration of formalin for helminths eggs and larvae
10%
It contains mercuric chloride (higly toxic to humans)
Schaudinn’s solution
PVA
Preserve fresh stool in preparation for staining stool smears
Schaudinn’s solution
Schaudinn’s solution contains what chemicals that is highly toxic to humans
Mercuric chloride
Plastic resin that serves as an adhesive for stool
PVA
Normally incorporated in Schaudinn’s solution
PVA
Stool preserved in those preservatives can be concentrated using FECT
PVA
Major drawback: use of mercuric chloride
PVA
Serves as staining agent in MIF preservatives
Merthiolate
Iodine
Acts as preservative in MIF
Formalin
Fixation of intestinal protozoan, helminth eggs and larvae
MIF
Component should always be fresh because it is unstable
Lugol’s iodine
Does not contain mercuric chloride
Sodium acetate-acetic acid formalin
Liquid fixative with a long shelf-life
Sodium acetate-acetic acid formalin
Images are not as sharp as PVA and Schaudinn’s solution after staining
Sodium acetate-acetic acid formalin
Characteristics of container
Clean, watertight,with tight fitting lid
Acceptable amount of stool
Formed: 2-5 g (thumb-sized/wallnut)
Liquid/Watery: 5-6 tbsp
Time collection of liquid stool
30 mins
Time collection of soft or semi-formed stool
1 hour
Time collection of formed stool
up to 24 hours
Fixative to stool ratio
3:1
Specimen must be fixed in the preservative for at least:
30 minutes before processing
Ova and parasite samples collected
3 samples in 10 days (intermittently)
How many samples are used for diagnosis of amebiasis
6 samples in 14 days
Patients undergoing barium,bismuth or mineral oil therapy
7-10 days before collection
Patients undergoing _____,_____, or _______ must have 7-10 days before collection of stool
Barium
Bismuth
Mineral oil therapy
Patients taking antibiotics or antimalarial drugs, gallbladder dyes
Delayed for 2 weeks
5 specimen used in parasitology
Stool
Blood
Urine
Sputum or BAL
Cerebrospinal fluid
Type of blood which is best for demonstrating blood parasites
Capillary blood
In processing blood parasites, avoid using _______
Anticoagulated blood
If EDTA blood is used, specimen must be processed within
1 hour
If _____ blood is used, specimen must be processed within 1 hour
EDTA
Time of collection in Malaria
Before the next anticipated fever spike
Time of collection in Filariasis
Depend on the periodicity of filariae
Specimen that useful of T. vaginalis trophozoites, S. haematobium eggs,rarely W. bancrofti adult
Urine
Specimen useful in demonstration of P. westermani eggs, E. histolytica trophozoites, migrating larvas of Ascaris, Strongyloides, and Hookworms, and P. jirovecii
Sputum or BAL
Urine is useful in demonstration of
T. vaginalis trophozoites (most frequent parasite)
S. haematobium eggs
W. bancrofti adult ( in cases of hydrocele) rarely
Specimen useful in demonstration of Naeglaria and Acanthamoeba trophozoites, Trypanosomes and Panstrongylus larvae
Cerebrospinal fluid
Sputum or BAL is useful in demonstration of
P. westermani eggs
E. histolytica trophozoites (in cases of pumonary abscess)
Migrating larvas of Ascaris
Strongyloides
Hookworms
P. jirovecii
10% formalin formalin + stool + 10% HCl strained in a gauze
Acid-ether concentration technique
CSF is useful in demonstration of
Naeglaria
Acanthamoeba trophozoites
Trypanosomes
Panstrongylus larvae
Intradermal test for T. spiralis
Bachmann Test
For detection of Strongyloides
Baermann Funnel
Wrap the sample in cheesecloth and submerge it in a funnel filled with water
Baermann Funnel
Immunologic test for T. spiralis and E. granulosus
Bentonite Flocculation Test
Bentonite Flocculation Test is used for detection of
T. spiralis
E. granulosus
Detects malarial aldolase
BinaxNOW test
Use in Acid-ether concentration technique
10% formalin
Stool
10% HCl strained in a gauze
Intradermal test for E. granulosus
Casoni test
Culture medium for T. cruzi
Chang’s medium
Chang’s medium is use to culture
T. cruzi
Casoni test is use to test
E. granulosus
For detection of Schistosoma spp
Circumoval Precipitin test
Circumoval Precipitin test is use to detect
Schistosomma spp
Patient serum + schistosomal eggs
Circumoval Precipitin test
Use in Circumoval Precipitin test
Patient serum
Schistosomal eggs
Culture medium for T. vaginalis
Diamond Medium
Feinberg-Whittington Medium
Diamond Medium is use to detect
T. vaginalis
Intradermal test for Leishmania spp.
Dr. Montenegro’s test
Dr. Montenegro’s test is used to detect
Leishmania spp.
Detects Giardia,Strongyloides, Clonorchis,Opisthorchis,Fasciola, and Cryptosporidium
Entero (string) test
Entero test is used to detect
Giardia
Strongyloides
Clonorchis
Opisthorchis
Fasciola
Cryptosporidium
Other term of Entero test
String test
Uses a gelatin capsule attached to a string
Entero (string) test
Egg hatching and sedimentation technique for detecting Schistosoma spp. eggs
Faust and Malloney’s Test
Faust and Malloney’s test is used for detecting
Schistosoma spp. eggs
Feinberg-Whittington Medium is used to detect
T. vaginalis
Most commonly used method for concentrating eggs, larvae and protozoan cysts
Formalin-Ether/ Ethyl Acetate Concentration Technique
10% formalin + stool — centrifuge—–> Ether/Ethyl acetate to the sediment
Formalin-Ether/ Ethyl Acetate Concentration Technique
Use in Formalin-Ether/ Ethyl Acetate Concentration Technique
10% formalin
Stool
Centrifuge
4 layers of Formalin-Ether
Ether/Ethyl acetate (Top)
Debris
Formalin
Sediment (Bottom, parasites)
Uses filter paper strip with water
Harada- Mori technique
Strongyloides larvae move upwards, and Hookworm larvae move downwards
Harada- Mori technique
In Harada- Mori technique this larvae move upwards
Strongyloides
In Harada- Mori technique this larvae move downwards
Hookworm
Uses wire mesh, newpaper, and cellophane soaked in glycerol-malachite green
Kato-Katz smear
For detection of microfilariae
Knott’s concentration
EDTA blood + 2% formalin — centrifuge—> Smear + Giemsa or Methylene blue
Knott’s concentration
Used in knott’s concentration
EDTA blood
2% formalin
Centrifuge
Smear
Giemsa/ Methylene blue
Detects histidine-rich protein II (HRP-II) antigen produced by P. falciparum
MalaQuick test
MalaQuick test is used to detect
HRP-II antigen
Parasite involve in MalaQuick test
P. falciparum
For detection of Trichinella (gold standard), T. solium, Spirometra and Sarcocystis
Muscle Biopsy
Muscle Biopsy is used to detect
Trichinella (gold standard)
T. solium
Spirometra
Sarcocystis
Culture medium for Leishmania spp and T. cruzi
Novy-MacNeal-Nicole (NNN) Medium
Now-MacNeal-Nicole (NNN) Medium is used to detect
Leishmania spp.
T. cruzi
Medium used for detection of Acanthamoeba spp.
Non-Nutrient agar with E. coli
Non-Nutrient agar with E. coli can detect what spp
Acanthamoeba spp.
Detects parasite LDH and differentiates the 4 Plasmodium species
Optimal Assay
For detection of Trypanosoma, Plasmodium and Wuchereria
Optimal Assay
Optimal Assay is use to detect
Trypanosoma
Plasmodium
Wuchereria
Uses fluorescent microscope and acridine orange fluorochrome
Quantitative Buffy Coat
Gold standard test for detection of T. gondii
Sabin-Feldman Dye Test
Sabin-Feldman Dye Test is used to detect
T. gondii
Tachyzoites + serum (Toxoplasma’s Ab’s) –> Distorted and colorless tachyzoites
Sabin-Feldman Dye Test
Preparation used in Sabin-Feldman Dye Test
Tachyzoites
Serum (Toxoplasma’s Ab’s)
Distorted and colorless tachyzoites
For detection of E. vermicularis eggs
Scotch Tape Swab
Scotch tape swab is used to detect
E. vermicularis eggs
Uses boiled sucrose solution with phenol
Sheather’s Sugar Flotation Technique
For coccidian oocyst (Cryptosporidium, Cyclospora, Cytoisospora), and B. coli cyst
Sheather’s Sugar Flotation Technique
Skin biopsy for detection of O. volvulus
Skin snip
Stain for rapid processing of thick smears, especially in malaria
Field’s stain
Stain for diagnosis of malarial thick and thin smear
Giemsa stain
Stain for SAF-, formalin-preserved, fresh and unpreserved stools
Iron hematoxylin
Stain for demonstration of uterine branches of Taenia spp.
India ink
Stain for bone marrow samples and blood flagellates
Leishman’s stain
Stain for emulsifying agent for stool and stains nuclei of protozoans
Lugol’s iodine
Stain for coccidian parasites
Modified acid-fast stain
Recommended stain for intestinal microsporidia
Modified Trichome stain
Stain for fresh and unpreserved stools
Trichome stain
Stain for blood cell morphology, but must be confirmed by Giemsa
Wright’s stain
0.1 M NaOH + stool –> counted in an ordinary slide
Stoll Egg count
Stoll Egg count uses
0.1 M NaOH
Stool
For screening malarial parasites (quantitation and identifying pigments)
Thick Blood film
For identifying malarial species
Thin Blood film
Routine O&P examination
Wet Mount (NSS, Iodine)
Iodine stains cysts yellow-brown but kills trophozoites
Wet Mount
In Wet Mount. iodine stain cysts what color
Yellow-brown
In Wet Mount, iodine stains cysts but kills what
Trophozoites
In Wet mount, this are not stained
Chromatoid bodies
Animal test for detection of T. spiralis, T. cruzi, W. bancrofti
Xenodiagnosis
Xenodiagnosis is used to detect what parasite
T. spiralis
T. cruzi
W. bancrofti
What parasite is detected in xenodiagnosis when albino mice is used
T. spiralis
What parasite is detected in xenodiagnosis when reduviid bugs is used
T. cruzi
What parasite is detected in xenodiagnosis when mosquitoes is used
W. bancrofti
For protozoan cysts and nematode eggs except Trichuris and Capillaria
Zinc Sulfate Flotation Technique
In Zinc Sulfate Flotation Technique, this parasites are not detected
Trichuris
Capillaria
Not appropriate for operculated eggs, Schistosomes, or infertile Ascaris eggs
Zinc Sulfate Flotation Technique
Specific gravity of ZnSO4
1.18
What are the 2 layer of Zinc Sulfate Flotation Technique
Supernatant (Top, parasites)
Liquid suspension (Bottom)
What is the epidemiologic triangle
Host
Environment
Agent
(HEA)
Order of Chain of Infection
Infectious Agent
Reservoir
Exit
Mode of Transmission
Entry
Susceptible host
(IREMES)
Man serves as what host in Echinococcus spp
Accidental IH
2nd IH of D. Latum
Small fish
2 Example of Facultative parasites
S. stercoralis
FLAs (Free-living amoebas) - Naegleria, Acanthamoeba spp
1 example of Transitory parasite
S. stercoralis (indirect cycle)
1 example of permanent parasite
H. nana - moxenous/homoxenous; only cestode
2 example of periodicity
Nocturnal
Diurnal
What is Heteroxenous
Requires 2 or more host
Example of zoonotic
B. coli
D. immitis
Agents of larva migrans (Toxocara spp)
What is Larva migrans?
Wondering larva
2 example of accidental host
Echinoccocus in man
Agents of Larva migrans
1 example of erratic parasites
Ascaris lumbricoides -cause ectopic ascariasis
2 example of spurious/coprozoic
Fasciola hepatica eggs - Infective stage is Metacercaria
Capillaria hepatica eggs
Transmitted by Reduviid/ Triatomine
Trypanosoma cruzi
Transmitted by Ixodes spp.
Babesia spp
Most common portal of entry
Mouth
Parasites transmitted through oral-anal intercourse
G. lamblia
C. Parvum
E. vermicularis
D. fragilis
E. histolytica
(G CEDE)
3 Parasites transmitted through skin penetration
- Hookworms
S. stercoralis - L3 larva/Filariform larva
Schistosoma spp. - Cercaria
1 example of parasites transmitted through skin inoculation(vector-borne)
Arthropods
Parasites transmitted through sexual intercourse
T. vaginalis
Parasites transmitted through intranasal
FLAs (N. flowleri and Acanthamoeba)
Parasites transmitted through vertical transmission
Plasmodium spp
Toxoplasma
Other agents such as syphilis,VZV
Rubella
CMV
HIV
(TORCH)
4 parasites that cause internal autoinfection
S. stercoralis
C. philippinensis
C. parvum
T. solium *
Cause reverse peristalsis, Tissue infection,Migrate to brain and fatal
T. solium
Cause external autoinfection
E. vermicularis - its egg is infective and contain fully develop larva
H. nana
Most common portal of exit
Anus
What phylum is both asexual reproduction and motile?
Sarcomastigophora
Ciliophora
What phylum has both sexual and asexual reproduction and non-motile
Apicomplexa
Reproduction of Sporozoans and coccidians ?and is it motile or non-motile
Sexual and Asexual
Non-motile
Reproduction of Amoeba,Flagellates,Ciliates?is it motile or non-motile?
Asexual
Motile
Not protozoans but consider stramenophiles
Blastocystis
2 example of microsporidians
Encephalitozoon spp
Enterocytozoon spp
Obligate intracellular fungi; smallest intestinal parasite (1.5-3 um)
Microsporidians
Size of Microsporidians
1.5 - 3 um
Atypical fungus
P. jirovecii
Nematodes (roundworms) are considered
Hermaphroditic/ Non-hermaphroditic?
Non-hermaphroditic (sexes are separate)
Trematodes are considered
Hermaphroditic/ Non-hermaphroditic?
All are Hermaphroditic(monoecious flukes/sexes are combined)
except Schistosomes (Non-Hermophroditic/ Diecious flukes)
Cestodes are considered
Hermaphroditic/ Non-hermaphroditic?
All are hermaphroditic (sexes are combined)
2 example of 2nd IH of P. westermani
- Crabs and Crayfish
IH of D. medinensis and G. spinigerum
Copepods
1st IH of D. latum and spirometra
Copepods
Serve as fixative in MIF
Merthiolate
Most common delivered specimen in Parasitology because most parasite are intestinal
Stool
Clearance period of Antacids,antidiarrheals,barium,bismuth,laxatives,mineral oil therapy
7-10 days
Clearance period of antimicrobial agents
2-3 weeks
What may form in stool if clearance period of Antacids,antidiarrheals,barium,bismuth,laxatives is not followed?
Leave crystalline residues
Cause of antimicrobial agents in stool
Decrease the number of protozoans
Clearance period of gallbladder dyes
2 weeks
Avoid contamination of stool in urine because ______
water because ______
soil because _____
Urine - trophozoites is toxic to urine
Water - may dilute the spx
Soil- may contain foreign parasites
Capacity of stool container
1/2 pint
Number of stool specimen examined?
3 spx collected separate days within 10 days (every other day)
Reason why does it need 3 spx examined on separate days?
To recover parasites that shed diagnostic forms intermittently (such as G. lamblia and S. stercoralis)
Liquid/Watery stool may contain what type of parasite
Trophozoite
Soft/Semi-formed stool may contain what type of parasite
Mixture of Trophozoite and Cysts
Formed stool may contain what type of parasite
Cysts
Why does water stool should be process immediately?
Trophozoite is sensitive to environmental change
Stool must be left at room temp,incubated, frozen
True or False
False - Not be left at …
Beyond 1 hour, stool must be ____ and its is short term preservation
Refrigerated
For longer period of stool preservation:
2 vial system
( 10% formalin & PVA)
Ratio of stool- preservative
- 1:3 ( 1:stool = 3: preservative)
10% Formalin can be performed in:
WT/CT/PS/IA?
All can be used except in Permanent staining
Stool fixative that can fix and stain
MIF
MIF cannot be performed in:
WT/CT/PS/IA?
Permanent Staining and Immunoassay(due to red color of merthiolate)
Stool fixative that can be used as permanent stain
SAF (Sodiuam acetate-acetic acid formalin)
SAF can be performed in:
WT/CT/PS/IA?
All of them can be performed
Stool fixative that contain Mercuric chloride
Schaudinns’ fluid
PVA
Schaudinn’s fluid do not contain
PVA
SF with PVA
PVA
Schaudinn’s fluid can be performed in :
WT/CT/PS/IA?
Permanent staining only
PVA can be performed in:
WT/CT/PS/IA?
Permanent staining only
Modified PVA was introduce to avoid hazard of mercuric chloride and can be used in
NAATs
PCR
Modified PVA contain
ZnSO4
CuSO4
Modified PVA can be performed in:
WT/CT/PS/IA?
Permanent staining only
Immunoassay need
( fresh/old stool? ) to detect ______
Fresh
E. histolytica
G. lamblia
C. parvum
Normal color of stool
Brown
Brown color of stool is due to
Stercobilin or Urobilin
Black stool is due to
Upper GI bleeding
Mucoid/Bloody stool is due to
Lower GI bleeding or dysentery cause by E. histolytica or B. coli
Normal consistency of stool
Soft to semi-formed
Gives clues on parasite stages present in stool
Consistency
Determine the sensitivity of egg-counting technique in stool
Consistency
In egg counting technique, the ___ the stool, the ____ the count
drier, higher
In egg counting technique, watery stool has ___ count
High/ low?
Low
In egg counting technique, Formed stool has ___ count
High/ low?
High
In O & P exam, this procedure can be performed
Wet mount
Concentration technique
Permanent staining
This O & P procedure has low sensitivity due to small amount of stool
Wet mount
Wet mount is AKA
Direct fecal smear
Wet mount is prepared by mixing ____ of stool with a drop of _____
2 mg
NSS
How many percent of NSS is used in wet mount
0.85%
O&P procedure that is useful in detection of motile trophozoites, helminths and larvae
Wet mount
Wet mount can be stained using
Lugol’s iodine/ D’ Antoni’s iodine - protozoan cysts and detecting glycogen inclusions
Nair’s Buffered Methylene Blue
- for trophozoites because troph can be killed by iodine
Concentration technique has ____ to parasite detection
Increase/Low sensitivity?
Increase sensitivity
2 classification of concentration technique
Sedimentation procedures
Flotation procedures
2 classification of sedimentation procedures
FECT - Formalin ether (ethyl acetate) concentration technique
AECT- Acid ether concentration technique
Most common sedimentation procedures
FECT
Preservative used in FECT
Formalin
In FECT, sediments may be stored in this
Formalin
In FECT, it is less hazardous and dissolve fats
Ether/ ethyl acetate
Sedimentation procedure that can be used with specimens preserved in formalin, MIF or SAF
FECT
Efficient in recovering most protozoan cysts, helminth eggs,operculate eggs and moderately effective in schistosome eggs
FECT
Less distortion of protozoan cysts compared to zinc sulfate flotation and AECT
FECT
Used fresh stool
AECT/ FECT
AECT
Sedimentation medium used in AECT
40% HCl
Used in AECT to precipitate and remove albuminous material and mucoidal substances
40% HCl
Recommended for animal parasites, Trichuris, Capillaria and Schistosoma eggs
AECT
Parasites may be lost to the plug of debris and possible destruction of protozoan cysts due to increase acidity
AECT
SG of parasite > SG of sedimentation medium
Sedimentation procedures - parasites settle at the bottom
Arrangement of sedimentation procedure in a tube
Ether/Ethyl Acetate –>
Plug of debris —>
Sedimentation medium (Formalin/ HCL) —>
Sediments (parasite if present)
Which is decanted in sedimentation procedure
Ether/Ethyl Acetate –>
Plug of debris —>
Sedimentation medium (Formalin/ HCL) —>
SG of medium > SG of parasite
Flotation procedure (parasite floats)
3 classifcation of flotation procedure
Zinc sulfate centrifugal flotation
Brine’s flotation
Sheather’s flotation
Arrangement of flotation procedure in a tube
Surface film(parasites present)
—>
Medium (Zinc Sulfate,HCl, Sucrose) —>
Sediment (may still contain parasites that failed to float)
Concentration of medium in Zinc sulfate centrifugal flotation
33% ZnSO4
SG of ZnSO4 centrifugal flotation
1.18 - fresh stool
1.2 - formalinized stool (need adjusted for parasites to float)
Unreliable for the recovery of nematode larvae, infertile eggs of Ascaris,eggs of Taenia,schistosomes and other cestodes or trematode eggs
-Not for heavy or dense eggs
Zinc sulfate centrifugal flotation
Medium used in Brine’s flotation
Saturated salt concentration
SG of Brine’s flotation
1.2
No centrifugation is required; not applicable for operculate and thin-shelled eggs because it might shrank due to high SG
Brine’s flotation
Medium of Sheather’s flotation
Boiled sugar solution/sucrose
Uses Boiled sugar solution/sucrose preserved with phenol
Sheather’s flotation
Sheather’s flotation is preffered for concentrating ____
coccidian oocysts
SG of Sheather’s flotation
1.25-1.27
Used for detecting coccidian
Sheather’s flotation and Modified acid fast stain
4 Permanent stains used in stool
Iron hematoxylin
Wheatley’s trichrome
Modified trichrome
Modified Acid fast stain
(IW MM)
Microscopic field used in Iron hematoxylin,Wheatley’s trichrome, Modified trichrome
300 OIFs
OIO = 100x objective
Microscopic field used in Modified Acid Fast stain
300 HPFs
(Red structure against blue background)
Iron hematoxylin has shade of
Gray blue to black
Wheatley’s trichrome is AKA
Gomori’s trichrome
Color of the ff in Gomori’s trichrome:
Cytoplasm:
Nucleus:
Inclusions (RBC,Bacteria):
Charcot Layden crystals
Cytoplasm: blue to green
Nucleus, Inclusions, Charcot Layden crystals : Red to Purple
Microscopic ojectives used in protozoa
OIO
Weaker acid decolorizer (1-3% H2SO4) as decolorizer; demonstrated Cryptosporidium,Cytoisospora,Cyclospora oocysts
Modified acid fast stain
Cryptosporidium
Acid fast/ Acid fast variable/Non-Acid fast?
Acid fast
Cytoisospora,Cyclospora oocysts
Acid fast/ Acid fast variable/Non-Acid fast?
Acid fast variable
2 example of Modified acid fast stain
Kinyoun’s method - Cold method; most common
Ziel-Neelsen method - Hot method
Artifacts that resemble parasites:
1.Neutrophils (Multilobed:
2. Macrophages,squamous and columnar epithelial cells:
3. Yeast, fungal conida, mushroom spores, diatomes:
4. Mite eggs:
5. Pollen grains:
- Mature E. histolytica cysts due to 4 nucleus
- Amebic trophoizotes
3.Protozoan cysts or helminth eggs - Hookworms eggs/ larva
- Taenia or Ascaris eggs
How many stool is usd in Kato-thick smear
50-60 mg
In Kato-thick smear the cellophane is soaked in a mixture of
Glycerin - clearing agent,dissolve debris, allow parasites
Malachite green - stain the background green;lower the light intensity of microscope
Usefull in mass stool examination and not easily dry up
Kato-thick smear
Used for assessing worm burden and monitoring the efficacy of antihelminthics in cases of STH and Schistosomiasis
Egg counting technique
Parasite involved in STH
Hookworms
Ascaris lumbricoides
Trichuris trichuria
(HAT)
Modification of kato-thick smear and stool has been sieved through mesh wire
Kato-Katz
Formula for egg counting technique
EPG (eggs per gram feces) = number of eggs counted x factor( depends on template used)
Standard template for egg counting technique
1.5mm thick and 6mm hole
Amount of feces delivered & factor in:
1.5 mm thick and 6 mm hole
1mm thick and 9 mm hole
0.5 mm thick and 6.5 mm hole
Amount of feces delivered & factor:
41.7 mg = 24
50 mg =20
20 mg = 50
Make used of calibrated flask and stoll pipette
Stoll dilution
Diluent used in stoll dilution
0.1 N NaOH
Used to determine the severity of Schistosoma infection
Faust-Malloney egg hatching technique
EPG of the following parasites in Light intensity
A. lumbricoides
T. trichuria
Hookworms
Schistosoma japanicum/mansoni
Light intensity:
A. lumbricoides = <5000
T. trichuria = <1000
Hookworms =< 2000
Schistosoma japanicum/mansoni = <100
EPG of the following parasites in Moderate intensity
A. lumbricoides
T. trichuria
Hookworms
Schistosoma japanicum/mansoni
Moderate intensity:
A. lumbricoides = 5000 -49,999
T. trichuria = 1000 - 9,999
Hookworms = 2000- 3,999
Schistosoma japanicum/mansoni = 100 - 399
EPG of the following parasites in Heavy intensity
A. lumbricoides
T. trichuria
Hookworms
Schistosoma japanicum/mansoni
Heavy intensity:
A. lumbricoides = ≥ 50,000
T. trichuria = ≥ 10,000
Hookworms = ≥ 4,000
Schistosoma japanicum/mansoni = ≥ 400
Stool culture method is routinely used and less hazardous
True/ False?
False
Not routine and hazardous
Stool culture method can be classified into 2
Copro culture
Harada-Mori or Test tube method
Uses moistened soil and granulated charcoal
Copro culture
In copro culture, larvae are harvested using what technique
Baermann technique
what method involves applying positive stool to a filter paper strip and placing it into a test tube with distilled water for ___ days at ____ °C
Harada-Mori or Test tube Method or Filter paper Method
10 days at 25-30 °C
Hookworm Vs. S. stercoralis
In Fresh stool?
In Old Stool?
Fresh stool:
Hookworms = eggs
S. stercoralis = Rhabditiform larva
Old stool:
Hookworms & S. stercoralis = Rhabditiform larva
Used to detect eggs of E. vermicularis
Perianal swab/ Scotch tape method/ Cellulose tape
E. vermicularis is
Nocturnal/Diurnal?
Nocturnal
Scotch tape method is performed at
Night?Morning?Afternoon?
Early morning because eggs deposition happens at night
In Perianal swab, a drop of ___ or __ may be added to improve the visualization of eggs
Xylene or Toluene
What procedure is used for diagnosis of amoebiasis (E. histolytica)
Sigmoidoscopy
Entero test is AKA
String test or Duodenal capsule
Entero test is used for the recovery of what parasites
G. lamblia
C. parvum
S. stercoralis
or other Intestinal parasites
Contact time of Entero test
4 hours
Collected material in Entero test must be analyzed within
1 hour
Prepared specimen for Malaria to avoid distortion of parasite morphology
Fresh
Non-anticoagulated blood/Capillary blood
Why does anticoagulated blood is not prepared for malarial examination
Can disrupt morphology of parasite and difficult to examine
Using anticoagulated blood (EDTA> Heparin) must be delivered to the laboratory within _____ and must be processed within _____
30 mins
1 hour
Timing of collection for Malaria
Before the next anticipated fever spike / at the height of fever
Timing of collection of filariasis
Depends on the periodicity of microfilariae
Gold standard for malarial examination
Stained smears of thick and thin blood smear
How many drops of blood for thick and thin smear
Thick smear - 2-3 small drops (6ul)
Thin smear - 1 drop (2-3ul)
Thick smear is dehomoglobinized using ____ prior to staining
distilled water
Why does thick smear need to be dehomoglobinzed?
For rbc to lyze and parasite will stand out and wbc will remain
Purpose of thick smear
Rapid diagnosis of malaria and demonstration of trypomastigotes and microfilariae
What type of smear, which is thick at one end and thin and feathery at the other end
Thin smear
Smear which is fixed with alcohol prior to staining
Thin smear
Smear for malarial species identification
Thin smear
Degree of parasitemia for Thick smear
Parasites/ul of blood = (parasites/WBCs counted) x WBC count per uL or 8,000 WBCs
How many parasites/ WBC need to be counted in thick smear?
500 parasites or 1000 WBC -whichever comes first
If WBC count is unkwown, assume 8,000 wbcs per ul of blood
Degree of parasitemia for thin smear
% parasitemia = (parasitized rbcs/ total RBCs counted) x 100
In thin smear, if parasitemia is high (>10%) examine ______ RBCS
If it is low (<1%) examine _____ RBCs
500
2000
4 Stains used in blood parasites
- Giemsa
- Wright’s
- Delafield hematoxylin
- Kawamoto technique
2 stains that are categorized as Romanowsky stain
Giemsa
Wright’s stain
Recommended for detection and identification of blood parasites (such as plasmodium & babesia); allows demonstration of stippling
Giemsa
Not optimal for blood parasites; does not demonstrate stippling
Wright’s stain
What stain that demonstrated detailed structure of microfilariae;requires dehomoglobinazation of thick films with _______
Delafied hematoxylin
2% formalin in 1% acetic acid
What stain are which blood smear are stained with ______ which results in a differential staining of nuclear DNA in _____ ( color)and cytoplasmic RNA in _____ (color)
Kawamoto technique
acridine orange
green
red
What artifacts resemble trypomastigotes or malarial stages
Elongated and degenerating platelets
What artifacts resemble plasmodium schizont
NRBCS
What artifacts resemble amastigotes of leishmania or T. cruzi
Yeast/ fungal elements
Uses capillary tube pre-coated with acridine orange which the parasites’ DNA takes up
Quantitative buffy coat (QBC)
QBC is pre-coated with
acridine orange
Type of microscope used in QBC and its positive color
Fluorescence microscope
Brilliant green
QBC is useful in demonstration of malarial parasites, microfilariae, trypomastigotes and babesia
True or False?
True
Concentration technique in blood specimen is useful in
Microfilaremia - microfilaria in blood
2 types of concentration technique in blood spx
Knott’s concentration
Membrane filtration
Knott’s concentration uses how many ml of blood?mixed with?
1 ml of blood mixed with 10 ml of 2% formalin followed by centrifugation
Sediments in Knott’s concentration is examined as a ____ or ____
Wet mount or stained smear
Membrane filtration uses how many ml of blood?lysed with?size of membrane filter?
1ml of blood, lysed with 10 ml of distilled water in a syringe with a 5 um membrane filter attached
Spx that is stat in parasitology
Blood and CSF
Spx for demonstration of Naegleria and Acanthamoeba trophozoites,trypomastigotes and cysticerci (T. solium larvae)
CSF
Spx used to detect T. vaginalis trophozoites,S. haematobium ova, rarely W. bancrofti microfilariae
Urogenital tract specimen
W. bancrofti usually found in ____
It may produce ____ urine
Hydrocele -swelling in the scrotum
Chyluric/Milky urine
Spx used for demonstration of P. westermani ova, migrating helminth larvae, E. histolytica trophozoites, and P. jirovecci
Sputum or BAL
In immunodiagbostic test, it is for detecting current infection
Antigen/Antibody/LFA/EIA?
Antigen detection
In immunodiagbostic test, it is for detecting past/chronic
Antigen/Antibody/LFA/EIA?
Antibody detection
Immmunodiagnostic test in which antibody is fluorescent labelled
DFA
Immmunodiagnostic test in which AHG is fluoresecent labelled
IFA
Preparation for SFDT
Px sample + Live T. gondii
–> incubate
–> Add Methylene Blue
–> Microscopic exam
(+) T. gondii in SFDT
Remain unstained - looses affinity to methylene blue when incubate and distorted
Preparationf for COPT
Px serum + Live schistosoma eggs
–> incubate
–> Microscopic exam
(+) result for COPT
Formation of blebs or separate precipitate around the eggs
Meaning of COPT
Circumoval precipitin test
Assay format and specimen used for:
E. histolytica
G. lamblia
C. parvum
LFA,DFA,EIA
Stool
Assay format and specimen used for:
T. vaginalis
LFA, DFA
Vaginal Swab
Assay format and specimen used for:
Plasmodium spp
P. falciparum
LFA
Blood
Target antigen in plasmodium spp for antigen detection
Parasite LDH
Target antigen for P. falciparum for antigen detection
HRP-2 (Histidine Rich Protein-2)
Assay format and specimen used for:
Leishmania spp.
IFA,EIA,CF
Serum
Assay format and specimen used for:
T. cruzi
IFA,EIA,CF,IB
Serum
Assay format and specimen used for: (in abs detection)
Plasmodium spp
IFA
Serum
Assay format and specimen used for:
T. gondii
IFA,EIA, SFDT
Serum/ CSF
Assay format and specimen used for:
W. bancrofti
EIA
Serum
Assay format and specimen used for:
T. spiralis
EIA, BF
Serum
Assay format and specimen used for:
Schistosoma spp
EIA,IB,COPT
Serum
Assay format and specimen used for:
T. solium (cysticercosis)
EIA,IB
Serum/ CSF
Assay format and specimen used for:
E. granulosus
EIA, IB, IHA, IFA
Serum/CSF
Molecular diagnostic which is highly sensitive and specific
PCR
Principle of PCR
NAATs (Nucleic Acid Amlification Test)
2 Types of PCR
Conventional PCR
Real time PCR
Involves detection of amplicons using gel electrophoresis
Conventional PCR
Real time PCR is AKA
qPCR (Quantitative PCR)
Uses fluorophores to enable detection of amplicons as they are produced
RT-PCR
Fluorophores used in RT-PCT
SYBR Green
Taq Man
Common target in PCR
rDNA
Steps in PCR
Denaturation
Annealing
Extension/Elongation
(DAE)
Separation of template DNA to single strands; Break hydrogen bonds
Denaturation
°C for denaturation
94 °C
Binding of primers specific for each target strand sequence
Annealing
°C for Annealing
50-65 °C
Have reverse and forward primer
Annealing
Addition of deoxyribonucleotides to produce new strands
Extension/Elongation
°C for Extension/Elongation in PCR
72 °C
Heat stable; from thermophilic bacteria
Taq polymerase
Only atrial amebae, does not have cysts, exist in trophozoite,
ingest WBCS
Entamoeba gingivalis
Only pathogenic amoeba, can lyse tissue, hematophacous
E. histolytica
Amoeba are mostly found in the
Intestine
Identical to E. histolytica, not capable of ingesting rbc
E. dispar/ E. moshkovski/E. bangladeshi
Small race of E. histolytica
E. hartmanni
Smallest intestinal amoeba
E. hartmanni
Endolimax nana
Amoeba which has 8 nuclei in its mature cysts
Entamoeba coli
Cross-eyed cyst, small/dwarf intestinal slug
Endolimax nana
Entamoeba which has 1 nucleus, large iodophilic mass/glycogen vacuole
Iodamoeba butschlii
Entamoeba which has 1 nucleus, non-iodophilic mass, parasite of pigs and monkeys
Entamoeba polecki
- Difference between E. histolytica and E. polecki
E. histolytica - 4 nucleus
E. polecki - 1 nucleus
Entamoeba cysts which has 1-4 nucleus ; its chromatoidal bodies is described as cigar-shaped
E. histolytica
E. dispar/E. moshkovski/E. bangladeshi
E. hartmanni
Entamoeba cysts which has 1-8 nucleus and its chromatoidal bodies is characterized as broom stick-splinter-like with pointed or rough ends
Entamoeba coli
Entamoeba cysts;4 nucleus; small,spherical,oval
E. nana
Hallmark of E. histolytica
Ingested rbcs
Entamoeba trophozoites; basket nucleus
Iodamoeba butschlii
2 Types of pathology in Amoebiasis
Intestinal Amebiasis
Extraintestinal Amebiasis/ Tissue Abscess
Intestinal Amebiasis can be
Acute/Symptomatic
Chronic/ Asymptomatic
Amoeboma
Amoebic ulcers
Cause amebic dysentery
Acute /Chronic?
Acute
Incubation period is ___ days; presents with bloody diarrhea and abdominal pain
Acute amebiasis
2-5 days
Cardinal sign of dysentery
Bloody diarrhea
Abdominal pain
Type of amoebiasis in which patients are considered as cyst passers/carriers; stool is soft to well-formed
Chronic
Carcinoma like or mass of granulomatous tissue in the intestine
Amoeboma
Flask-shaped ulcers due to hyaluronidase
Amoebic ulcers
Most common site in extraintestinal amebiasis
Liver (hepatic abcess)
Why does the liver is the most common site in extraintestinal amebiasis?
Proximity to large intestines
4 Examples of Free-Living Amoebae
N. flowleri
Acanthamoeba spp.
Balamuthia mandrillaris
Sappinia spp
N. fowleri is AKA
Amoeboflagellate
Trophozoite stage can transform into a flagellate stage or a resistant cyst; Biphasic
N. fowleri
Opportunistic free-living amoebae which include A. castellani, A. culbertsoni, A. polyphaga,A. hatchetti, A. rhysodes
Acanthamoeba spp
Morphologically similar to Acanthamoeba;
Cyst= trilayered
Trop=finger like pseudopods
Balamuthia mandrillaris
Rare cause of meningoencephalitis
Sappinia spp.
Not present in tissue,small,uninucleate,double- walled with smooth outer wall
N. fowleri cyst
- Present in tissue,large,uninucleate, double-walled with wrinkled-outer wall
-Ectocyst is wrinkled
Acanthamoeba spp cyst
2 biphasic form of N. fowleri
Amoeboid form
Flagellate form
Form of N. fowleri which has lobate pseudopods (lobopods)
Amoeboid form
Form of N. fowleri which has 2 anterior flagella
Flagellate form
Rapidly motile
N. fowleri/ Acanthamoeba spp?
N. fowleri
Slowly motile
N. fowleri/ Acanthamoeba spp?
Acanthamoeba spp
Characterized by spiny/filiform pseudopods (acanthopods); No flagellate form
Acanthamoeba spp
It cause PAM(Primary Amoebic Meningoencephalitis) acute
N. fowleri/ Acanthamoeba spp?
N. fowleri
It cause GAE (Granulomatous Amebic Encephalitis) chronic , keratitis, skin lesion
N. fowleri/ Acanthamoeba spp?
Acanthamoeba spp
Diagnostic stage of N. fowleri
Cyst/ Trop?
Trophozoites (amoeboid/flagellate)
Diagnostic stage of Acanthamoeba spp
Cyst/ Trop?
Cyst & Trophozoites
G. lamblia is AKA
G. duodenale
4 nuclei, parabasal bodies, remnants of flagella
G. lamblia
1 nucleus (lemon, nipple cyst), cytostome with curved fibril referred to as “shephered’s crook fibril”
C. mesnili
2 or 4 nucleus, bipolar arrangement of nuclei, resembles E. nana cyst
E. hominis
E. hominis resemble
E. nana cyst
1 nucleus, cystostome extends above the nucleus “ bird’s beak”
R. intestinalis
G. lamblia is found in
small/large intestine?
Small intestine
It is considered as atrial flagellates and have no cyst
P. hominis
T. tenax
T. vaginalis
Is is known as “old man with eyeglasses”
G. lamblia
2 nuclei, 8 flagella, prominents sucking disc and axostyle,
* only bilaterally symmetrical
G. lamblia
Motility of G. lamblia
Falling leaf-like
How many is the flagella of G. lamblia?
8 Flagella
C. mesnili, E. hominis, R. intestinalis, D fragilis. P. hominis is found in the:
small/ large intestine?
Large intestine
1 nuclei, 3 anterior flagella & 1 fibril,prominent cytostome with spiral groove
C. mesnili
Motility of C. mesnili
Cork-Screw/ Spiral
How many is the flagella of C. mensnili
3 Anterior flagella
1 nuclei, 4 flagella, one side of the body is flattened
E. hominis
Motility of E. hominis
Jerky
How many is the flagella of E. hominis
4 flagella =
3 Anterior flagella
1 Posterior flagella
1 nuclei, 2 flagella; cytostome extends 1/2 of the length of the body
R. intestinalis
Motility of R. intestinalis
Jerky
How many flagella of R. intestinalis
2 Flagella =
1 anterior flagella
1 posterior flagella
1 or 2 nuclei , no flagella, multiple hyaline,leaf-like pseudopods, fragmented tetrakaryosome
D. fragilis
1 nuclei, 5 flagella, undulating membrane; axostyle extends beyond the organism’s body, cytoplasmic granules
P. hominis
T. tenax
T. vaginalis
Motility of D. fragilis
Amoeboid
Motility of P. hominis, T. tenax,
T. vaginalis
Rapid, jerky, tumbling
How many is the flagella of T. vaginalis
5 flagella =
4 Anterior flagella
1 Posterior flagella
T. tenax is found in:
Vagina/ Urogenital/ small intestine/ large /intestine/mouth?
Mouth
T. vaginalis is found in
Mouth/ Intestine/ Urogenital tract?
Urogenital tract
Longest undulating membrane; have axostyle extend beyond the body ; as long as costa
Pentatrichomonads hominis/ P. hominis
Undulating membrane is almost as long as costa
T. tenax
Undulating membrane is 1/2 or 2/3 of costa; shortest undulating membrane
T. vaginalis
What are the trichomonads parasite?
P. hominis
T. tenax
T. vaginalis
Cause traveler’s diarrhea and has been associated with homosexual men with gay bowel syndrome; pathogenic
G. lamblia
G. lamblia use what type of test?
Entero test
Intermittent diarrhea,nausea,vomiting, abdominal pain; pathogenicity depends on parasite loads
D. fragilis
Diagnostic technique for D. fragilis
Trichrome stain
Cause non-specific vaginitis,urethritis, or prostatitis, foul smelling discharge (mucopurelent);always pathogenic
T. vaginalis
Gold standard medium for T. vaginalis
Diamond’s medium
Categorized as Hemoflagellates
Leishmania spp
Trypanosoma brucei complex (T. b gambiense, T. b. rhodisiense)
T. cruzi
What parasite?
Skin inoculation of promastigote by sand flies
Leishmania spp
Parasite involves in Phlebotomus fly
Leishmania spp
Parasite involve in Lutsomyia fly
Leishmania spp
Diagnostic stage of Leishmania spp or in Man
Amastigote (intracellular replication form)
What parasite?
Skin inoculation of metacyclic trypomastigote by Tse-tse flies
Trypanosoma brucei complex
Diagnostic stage of T. brucei complex
Trypomastigote (extracellular)
What genus is involve in Tse-tse fly?
Glossina spp
What parasite?
Indirect inoculation of metacyclic trypomastigote from feces of reduviid/Triatomine bug/ Kissing bug?
T. cruzi
Vector involve in T. cruzi?
Reduviid
Triatomine
Kissing bug
Diagnosic stage of T. cruzi
Amastigote & Trypomastigote
Developmental stage of Hemoflagellate
Promastigote
Amastigote
Metacylic Trypomastigote
Epimastigote
Trypomastigote
(PAMET)
Infective stage in sandflies/ Leishmania?
Promastigote
Culture medium used to grow Leishmania
Novy-MacNeal-Nicolle (NNN) or
Schneider’s Orosophilia medium
Infective stage of T. cruzi and T. brucei
Metacylic trypomastigote
Occurs in vectors; short undulating membrane
P/A/M/E/T?
Epimastigote
Full undulating membrane due to kinetoplast; diagnostic stage in man; C,S, or U shaped in stain thick blood fim
Trypomastigote
What is the life cycle of Leishmania spp
Promastigote (infective stage-sand fly) –> Amastigote (diagnostic stage-man)
What is the life cycle of T. brucei
Metacyclic Trypomastigote (infective stage is in the tse-tse fly; infective form is in the salive)
—> Trypomastigote (diagnostic stage in man)
–> Epimastigote (Tsetse fly)
—> Metacyclic Trypomasigote
(MET)
What is the life cycle of T. cruzi?
Metacyclic trypomastigote (IS)
–>Amastigote (DS in man)
—>Trypomastigote (DS in man)
–> Epimastigote (hindgut)
—> Metacylic trypomastigote
(AMET)
3 classification of Leishmaniasis
Cutaneous
Muco-cutaneous
Visceral
Other term for Old World CL
Oriental sore
Delhi ulcer
Aleppo boil
Baghdad boil
Other term for New World CL
Chiclero’s ulcer
site: earlobe
Other term for Old World VL
Dumdum fever
Black fever
* Kala-azar
Darkening of the skin;
vector: sandfly
Kala-azar
Agent of old world CL
L. tropica
L. major
Agent of new world CL
L. mexicana (transmitted by Lutzomyia fly)
Skin lesions with elevated margins that leave ugly scars on healing
Cutaneous Leishmaniasis
Specimen for diagnosis in CL
Skin biopsy
Cause espundia or tapir nose
Muco-cutaneous leishmaniasis
Causative agent of Muco-cutaneous leishmaniasis
L. braziliensis
Leprosy-like cutaneous lesions that spread to the mucous membranes
Muco-cutaneous Leishmaniasis
Spx for diagnosis of Muco-cutaneous leishmaniasis
Skin and Mucosal biopsy
Old world VL
L. donovani
New world VL
L. chagasi
Fever, organomegaly, ascites,anemia, hyperpigmentation of the skin
Visceral Leishmaniasis
Spx for diagnosis in VL
Tissues aspirate
Biopsy
African Trypanosomiasis is AKA
Sleeping sickness
Causative agent of West African Trypanosomiasis
T. b. gambiense (chronic)
Causative agent of East African Trypanosomiasis
T.b. rhodesiense (acute)
3 Manifestation of Sleeping sickness
- Trypanosome chancre - lesion on the bite site
- Hemolymphatic stage - febrile stage or cervical lymphanopathy (Winterbottom’s sign)
- Meningoencephalitic or sleeping stage - somnolence (daytime sleepiness), coma, death
Causa Winterbottom’s sign
African trypanosomiasis
Spx for diagnosis in sleeping sickness
Blood
Lymph node aspirate
CSF
New World or American Trypanosomiasis is AKA
Chagas disease
Causative agent of Chagas disease
T. cruzi
Causative agent of New World or American Trypanosomiasis
T. cruzi
Lesions at the inoculation site of Chagas disease
Romania - swelling around the eyes (orbital edema)
Chagoma -swelling at other sites
Largest intestinal protozoan
B. coli
Common parasite of pigs (zoonotic)
B. coli
B. coli is a
ciliate/apicomplexa/sarcomastigophora?
Ciliate
Habitat of B. coli
Large intestine
MOT of B. coli is:
Ingestion of ____
Sarcocysts/spores/oocysts/cysts?
Cysts
Immediately infective in C. parvum
cysts/oocyst/sarcocyst?
oocysts
Cause cryptosporidiosis
C. parvum
C. parvum, C. beli, C. cayetanensis,S. hominis,S. suihominis,S. lindemanni is classified as:
Ciliate/ Microscoporidians/ Apicomplexans/Stramenophiles?
Apicomplexans
Cause cytoisosporiasis
C. beli
Cause cyclosporiasis
C. cayetanensis
MOT of C. parvum, C. beli, C. cayetanensis
Ingestion of _____
Mature oocysts/Immature oocysts/cysts/spores
Mature oocysts
MOT of S. hominis
Ingestion of _____ from undercooked meat
sporocysts/sarcocysts/spores/
cyst?
Sarcocysts
Cause intestinal sarcocystosis
S. hominis
Habitat of S. lindemanni
Muscle
Definitive host and intermediate host of S. lindemanni
DH: Dog and Cat
IH:Man serve as dead-end intermediate host
Cause tissue sarcocystis
S. lindemanni
Blastocystis hominis is considered?
Thermophiles/Aerophiles/Stramenophiles
Stramenophiles
MOT of S. lindemanni
Accidental ingestion of ___
Thick walled cyst/ mature oocyst/ spores?
Mature oocyst
E. intestinalis and E. bieneusi is considered
Microscoporidians
MOT of E. intestinalis and E. bieneusi
Ingestion or inhalation of ___
Cysts/ spores/ oocyst
Spores
Cytopyge of B. coli trophozoite serve as?
Anus
C. parvum is considered?
modified acid fast/modified acid fast variable?
what color?
Modified acid fast
Pink to red
Diagnostic stage of C. beli and C. cayetanensis
Immature oocyst
Infective stage of C. beli and C. cayetanensis
Mature oocyst
C. beli and C. cayetanensis
Modified acid fats/ Modified acid fast variable?
Modified acid fast variable
Cause (+) auto florescence
C. beli
C. cayetanensis
S. hominis
Color of S. homonis in autofluorescence
Bright blue
Nuclei of B. homis is reported atleast
≥ 5/hpf
Type of microscope used in Microscoporidia
Electron microscope
Stain used in Microscoporidia
what color?
- Modified trichrome - pink to red or diagonal stripe
- Calcofluor white - bluish white fluorescence
Formation of oocyst from the union
Sporogonony
DH of malarial parasites
Female anopheles mosquito
IH of malarial parasites
Man or other vertebrae
Formation of schizonts
Schizogony/Merozogony
Formation of gametocytes
Gametozogony
Sleeping or dorminant;
When activated can cause relapse or malarial symptoms
Hypnozoites
3 cycle in Malarial parasites
Exo-erythrocytic cycle
Erythrocytic cycle
Sporogonic cycle
Exo-erythrocytic cycle happens in
Liver(hepatic)
Mosquito takes a blood meal( injects ______)
sporozoites
Mosquito takes a blood meal ( ingests ______)
gametocytes
Infective stage of malarial parasites
sporozoites
Diagnostic stage of malarial parasites
Immature trophozoite (ring stage)
Mature trophozoite
Gametocytes
Explain the life cycle of Malarial parasites
-Mosquito injects sporozoites (IS)
-Hypnozoites (when activated , cause relapse malarial symptoms)
Exo-erythrocytic cycle:
-Infect the liver cell
-Schizont
-Ruptures schizont (Merozoites)
Erythrocytic cycle:
-Human blood stages
-Immature trophozoites (ring stage)–> Gametocyctes = P. falciparum)
-Immature trophozoites –> Mature trophozoites–> schizont–>.ruptured schizont –> gametocyctes (P. vivax,P.ovale.P.malariae)
Sporogonic cycle:
-Mosquito injest gametocyctes
-Microgametocyte entering the Macrogametocyte
-Ookinete (zygote)
-Oocyst
-Release of sporozoites
Age of RBC infected:
P. falciparum:
P. vivax:
P.ovale:
P.malariae:
P. falciparum: All ages
P. vivax: Young rbc
P.ovale: Young rbc
P.malariae: Mature rbc
Appearance of parasitized rbc:
P. falciparum:
P. vivax:
P.ovale:
P.malariae:
P. falciparum: Normal,multiple infection of rbc, 8 ring forms
P. vivax: Larger than normal
P.ovale: Larger than normal; fimbriated edges
P.malariae: Normal in size or shape or maybe slightly smaller
RBC stippling in malaria, using Giemsa stain:
P. falciparum:
P. vivax:
P.ovale:
P.malariae:
P. falciparum: MAurer’s dots
P. vivax: Schuffner’s dots
P.ovale: Schuffner’s dots/ James’s dots
P.malariae: Ziemann’s dots
Stages in the peripheral blood :
P. falciparum:
P. vivax:
P.ovale:
P.malariae:
P. falciparum:
Ring, Gametocytes
P. vivax, P.ovale, P.malariae :
Ring, Mature trophozoites, Schizonts, Gametocytes
Length of erythrocytic cycle:
P. falciparum:
P. vivax:
P.ovale:
P.malariae:
P. falciparum: 36-48 hrs
P. vivax: 48 hrs
P.ovale: 48 hrs
P.malariae: 72 hrs
Shortest erythrocyctic cycle in malaria
P. falciparum
Longest erythrocytic cycle in malaria
P. malariae
Disease caused:
P. falciparum:
P. vivax:
P.ovale:
P.malariae:
P. falciparum: Malignant tertian malaria
P. vivax: Benign tertian malaria
P.ovale: Ovale tertian malaria
P.malariae: Quartan malaria
Most fatal form of malaria
P. falciparum
Rarely found in the peripheral blood of P. falciparum because they are sequestered in the deep vasculature (small blood vessels of internal organs) where schizogony occurs
Mature trophozoites
Schizonts
Developmental stage of malaria:
*multiple infection of rbc up to 8 per cell
*Double chromatin dots
-Applique/ accole form/ marginal
Young trophozoites of P. falciparum
Developmental stage of malaria:
-RBC are oval in shape
-Ringform is dense and compact
Young trophozoite in P. ovale
Malarial parasites that has band form
P. malariae
Malarial parasites that has fimbriated rbc
P. ovale
In what stage does stippling of malarial parasites start to appear
Mature trophozoites
32 merozoites
P. falciparum
12-24 merozoites
P. vivax
8-12 merozoites
P. ovale
- Malarial parasites arrange in rossette or daisy formation or fruit pie arrangement
P. malariae
6-12 merozoites
P. malariae
Microgametocytes: Sausage shape, scattered chromatin
P. falciparum
Macrogametocytes: Crescent shape with pointed ends, central chromatin mass
P. falciparum
Microgametocytes: Central chromatin mass
P. vivax. P. ovale. P. malariae
Macrogametocytes: Peripheral chromatin mass, round/ oval
P. vivax. P. ovale, P. malariae
Sausage shape, scattered chromatin in P. falciparum
Macrogametocytes/ Microgametocytes?
Microgametocytes
Crescent shape with pointed ends, central chromatin mass
Macrogametocytes/ Microgametocytes?
Macrogametocytes
Central chromatin mass in P. vivax, P. ovale, P. malariae
Macrogametocytes/ Microgametocytes?
Microgametocytes
Peripheral chromatin mass in P. vivax, P. ovale, P. malariae
Macrogametocytes/ Microgametocytes?
Macrogametocytes
Incubation period of Malaria
8-40 days
Classical sign and characterized by chills, fever, and profuse sweating
Paroxysm
Periodicity of malaria is determine by
length of the erythrocytic cycle/ recurrence of paroxysm
Periodicity of P. knowlesi
Quotidian - every 24 hrs
Periodicity of P. falciparum, P. vivax, P. ovale
Tertian malaria - every 48 hrs
Periodicity of P. malariae
Quartan- every 72 hrs
What type of Malarial parasites:
Manifestation:Intravascular hemolysis leading to Black water fever, anemia, cerebral malaria
P. falciparum
Black water fever
P. falciparum
Relapse may occur with ______ infections due to reactivation of ____ in the liver
P. vivax
P. ovale
Hypnozoites
What type of malarial parasites?
Sickle cell trait has resistance to
P. falciparum
Blood type group that has resistance to P. vivax
Duffy- negative blood group - common among blacks
5th human malaria
P. knowlesi
Causative agent of Simian malaria
P. knowlesi
Type of malaria that parasitize monkey
Simian malaria
P. knowlesi:
Young forms resemble___
Mature forms resemble ___
P. falciparum
P. malariae
Common encountered Babesia
B. microti
Intraerythrocytic parasites that resembles plasmodium spp
Babesia spp
T. gondii _____ are resistant to freezing, drying and extreme environmental conditions but are killed by heating to _____°C for ___ mins
Oocysts
70°C
10 mins
Babesia spp
DH:
IH:
DH:Ticks (Ixodes)
IH: Rodents ; Man -accidental IH
T. gondii
DH:
IH:
DH: Cat
IH: Rodents & Bird ; Man- accidental IH
Babesia spp
Skin inoculation of ______
by ticks
sporozoites
T. gondii
Ingestion of _____ from infected meat
Ingestion of ____ from cat’s feces
tissue cysts
oocysts
Pear shaped ring forms, tetrads(maltese cross)
Babesia spp
Hallmark of babesiosis
Maltese cross
The rapidly dividing form of T. gondii that causes acute infections.
Tachyzoites
The slowly growing form of T. gondii that causes chronic infections.
Bradyzoites
Babesiosis is AKA
Piroplasmosis
Babesiosis in cattle can cause
Red water fever
Most frequent parasite in urine
T. vaginalis
