PARASITOLOGY | Rocelle Anne R.

Leonardo, RMT

PART I. INTRODUCTION

PARASITE-HOST RELATIONSHIP

TYPE OF PARASITE
Obligatory parasite Parasite that cannot survive outside of a host
Facultative Parasite that is capable of existing independently of a host
Endoparasite Parasite that is established inside of a host
Ectoparasite Parasite that is established in or on the exterior surface of a host
TYPE OF HOST
Accidental or incidental host Host other than the normal one that is harboring a parasite

Definitive host Host in which the adult sexual phase of parasite development occurs
Intermediate host Host in which the larval asexual phase of parasite development occurs
Reservoir host Host harboring parasites that are parasitic for humans and from which humans may become infected
Transport host Host responsible for transferring a parasite from one location to another
Carrier Parasite-harboring host that is not exhibiting any clinical symptoms but can infect others
PARASITE-HOST RELATIONSHIP TERMS
Symbiosis Living together the association of two living organisms, each of a different species

Commensalism Association of two different species of organisms that is beneficial to one and neutral to the other
Mutualism Association of two different species of organisms that is beneficial to both
Parasitism Association of two different species of organisms that is beneficial to one at the other’s expense
Commensal Relating to commensalism; the association between two different organisms in which one benefits and has a neutral
effect on the other
Pathogenic Parasite that has demonstrated the ability to cause disease

PARASITE LIFE CYCLE Most commonly observed symptoms: diarrhea, fever, chills, abdominal pain, and abdominal
3 Components: cramping.
1. Mode of transmission
2. Infective stage: the morphologic form that invades humans PART II. SPECIMEN COLLECTION AND HANDLING
3. Diagnostic stage: one (or more) forms that can be detected via laboratory methods
Stool for Ova and Parasite Examination
DISEASE AND SYMPTOMS
- most common procedure in parasitology
The major body areas associated with such processes include the following:
- 2 general components:
1. gastrointestinal and urogenital tracts
o Macroscopic
2. blood and tissue
o Microscopic
3. liver, lung, and other major organs
4. miscellaneous locations, such as cerebrospinal fluid, eye, skin, and extremities.

1
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Collection - Semiformed stools: within 1 hour

- Typical protocol: 3 specimens, 1 specimen collected every other day or a total of 3 - Formed stool: within 24 hours
collected in 10 days.
- In cases of amebiasis: six specimens in 14 days is acceptable. Precautions:
- Stool samples from patients whose therapy includes barium, bismuth, or mineral oil should - Urine should not contaminate the stool specimen because it has been known to destroy
be collected prior to therapy or not until 5 to 7 days after the completion of therapy. some parasites.
- Stool samples from patients who have taken antibiotics or antimalarial medications should - Stool should not be retrieved from toilet bowl water because free-living protozoa and
be delayed for 2 weeks following therapy. nematodes may be confused with human parasites. Water may destroy select parasites,
- Type of container: clean, watertight container with a tight-fitting lid. such as schistosome eggs and amebic trophozoites.
- Acceptable amount of stool required: 2 to 5 g (or walnut-size) - Toilet paper in the stool specimen may mask parasites or make examination of the sample
difficult.
Transport
- The specimen container should be labeled with the patient’s name and identification
number, the physician’s name, and the date and time of sample collection. Fixatives for Collection
- Some form of requisition, paper or computer-based, should accompany the specimen - Freshly collected stool is the ideal specimen
indicating the test(s) requested. Suspected diagnosis, travel history, and clinical findings, is - If not possible, sample must be preserved to maintain its integrity
helpful, but may not be provided. - Ratio of fixative to stool – 3:1
- The specimen should be placed into a zip lock plastic bag for transport to the laboratory. - The specimen must be fixed in the preservative for at least 30 minutes before processing
Separate the paperwork from the specimen. begins.
- Liquid stool: within 30 mins

Advantages Disadvantages
Formalin - All-purpose fixative for the recovery of protozoa and  easy to prepare  Potential health hazard
helminths.  it preserves specimens for up to several  it does not preserve parasite morphology adequately
- Two concentrations commonly used: years for permanent smears
 5% ideally preserves protozoan cysts  it has a long shelf life.  trophozoites usually cannot be recovered
 10% preserves helminth eggs and larvae.  morphologic details of cysts and eggs may fade with
- direct examinations ✔ time
- concentration procedures ✔
- permanent smears ✗
Polyvinyl Alcohol - comprised of a plastic powder that acts as an adhesive for  Trophozoites and cysts of the protozoa,  Potential health hazard due to presence of mercury in
(PVA) the stool when preparing slides for staining. most helminth eggs, may be detected Schaudinn sol’n
- often combined with Schaudinn solution (zinc sulfate,  used for preparation of a permanent
copper sulfate, or mercuric chloride as a base) stained smear
- two-vial system:  Can be used for concentration techniques
 formalin vial for the concentration technique but not as effective
 PVA vial for the stained slide  Has a long shelf life when stored at room
temperature

2
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Sodium Acetate - alternative to PVA and Schaudinn fixative  only requires a single vial  adhesive properties are not good (add albumin to the
Formalin (SAF) - used for preparing smears for staining with the modified  mercury-free slide)
acid-fast stain to detect coccidian oocysts.  easy to prepare and long shelf life  protozoa morphology is not as clear in permanent
stains as when mercury-containing preservatives are
used
Modified PVA - alternatives to mercury-based PVA  used for concentration methods and  does not provide the same quality of preservation for
- uses copper sulfate or zinc sulfate permanent stained smears protozoan morphology on a permanent slide as the
mercury-based fixatives. Zinc sulfate fixatives > copper
sulfate.
Alternative Single- -  free of formalin and mercury  Does not provide the same quality of preservation as
Vial system  can be used for concentration techniques mercury-based fixatives and organism identification
and permanent stained smears. will be more difficult from permanent stained
 used for performing fecal immunoassays slides.

Processing  Direct saline wet preparation

1. Macroscopic Examination  a drop of 0.85% saline + small portion of unfixed stool using a wooden
- Consistency or degree of moisture applicator stick on a glass slide.
 soft or liquid stools - protozoan trophozoites  A 22-mm square cover slip is placed on the slide and the preparation is
 fully formed stools - protozoan cysts examined microscopically in a systematic fashion using the low power (10×)
 liquid or formed stools - Helminth eggs and larvae objective
- Color  A temporary seal can be prepared using a hot paraffin-petroleum jelly
 may indicate the condition of the patient, such as whether a patient had a recent (Vaseline) mixture around the edges of the cover slip.
special procedure (e.g., a barium enema) or if the patient is on antibiotic therapy
 Brown – normal  Direct iodine wet preparation
- Gross abnormalities  made to enhance the detail of protozoan cysts
 adult worms, proglottids, pus, and mucus  uses a drop of iodine (Lugol’s or D’Antoni’s formula) in place of saline
 Sample should be broken up using wooden applicator stick  Because iodine kills trophozoites present, it is recommended to use direct
 Samples containing adult worms may be carefully washed through a wire screen saline and direct iodine wet preparations on each sample

2. Microscopic
- Should be performed on fresh specimen; if received in fixative, the direct wet preparation  Concentrated Wet Preparations
can be eliminated from the O&P procedure  2nd procedure in an O&P examination
- Involves three procedures:  provide the ability to detect small numbers of parasites that might not be detected
 Direct Wet Preparation (direct wet mount) using direct wet preparations
 slide made by mixing a small portion of unfixed stool with saline or iodine  can be performed on fresh or preserved stool
 for detection of motile protozoan trophozoites. Protozoan cysts, oocysts,  allows detection of protozoan cysts, oocysts, helminth eggs, and larvae. Protozoan
helminth eggs, and larvae can also be observed trophozoites do not usually survive the procedure.

3
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

 2 types of concentration methods:  The slides are reviewed under OIO (100×); 300 fields are reviewed before the
 Formalin–Ethyl Acetate Sedimentation slide can be considered negative.
 The most widely used sedimentation technique; based on specific gravity  Two common stains used:
 3mL of ethyl acetate is added to formalin-fixed (9 mL of 10% solution) sample  Wheatley Trichrome
 Parasites – heavier than the solution and settle in the sediment; fecal debris –  Most widely used permanent stain
lighter and rises to the upper layer  Has long shelf life and easy to perform
 Advantage: provides good recovery of most parasites and is easy to perform  Preferably PVA-fixed specimen
 Disadvantage: preparation contains more fecal debris than a flotation  Iron Hematoxylin
technique  Reveals excellent morphology of intestinal protozoa
 Zinc Sulfate Flotation  In some cases, the nuclear detail of these organisms is considered to be
 based on specific gravity; sample debris (heavier) which sinks to the bottom stained clearer and sharper than when stained with trichrome
of the tube, and parasites (lighter) which float toward the top  Preferably SAF-fixed specimen
 Zinc sulfate (SG: 1.18-1.20) is added to the specimen and centrifuged.  Specialized stains:
 Advantage: more fecal debris is removed and it yields a cleaner preparation  Modified Acid-Fast Stain
 Disadvantage: some helminth eggs are very dense and will not float;  Staining procedure for detection of oocysts of Cryptosporidium, Isospora,
therefore, some parasites will be missed. and Cyclospora
 Sheather’s Sugar Flotation  Modified iron hematoxylin
 For recovery of coccidian oocysts (Cryptosporidium, Cyclospora, Isospora)  Incorporates carbol fuchsin step
 Boiled sugar solution preserved with phenol  Allows for the detection of acid-fast parasites in addition to the other
protozoa normally recovered using the iron hematoxylin stain
Sedimentation Schistosoma, Operculated eggs, Trematode egs, Cestode eggs, Trichuris  Uses SAF-preserved fecal samples
trichiura eggs, and Capillaria philippinensis  Modified Trichrome
Flotation Protozoan cyst, Nematode eggs except T. trichiura eggs and Capillaria  Enhanced detection of microsporidia
philippinensis

 Permanently Stained Smear OTHER INTESTINAL SPECIMENS:

 Final procedure in the O&P exam
 Microscope slide containing a fixed sample that has been allowed to dry and
subsequently stained, cover-slipped and sealed.
 Allows observation of detailed features of protozoa by staining intracellular
organelles
 Identification is considered tentative until confirmed with the permanent stained
smear
 Not the method of choice for the identification of helminth eggs or larvae because
these parasites often stain too dark or appear distorted.
 Helminth eggs or larvae are best detected and identified using a concentration
technique.
 Sample of choice: PVA-preserved sample
1. Duodenal material
- Collected by nasogastric intubation or by the enteric capsule test
- Parasites that may be observed: Giardia intestinalis trophozoites, Cryptosporidium spp.,
Isospora belli, Strongyloides stercoralis , and eggs of Fasciola hepatica or Clonorchis sinensis
- Enterotest: simpler method for collecting duodenal material without requiring
intubation.
 The patient swallows a gelatin capsule that contains a coiled length of yarn. The
capsule dissolves in the stomach and the weighted string is carried to the duodenum.
 After a 4-hour incubation period, the yarn is pulled back out of the patient. The bile-
stained mucous material brought up on the string is then examined microscopically
via wet preps

4
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

2. Sigmoidoscopy material  Knott technique

- Helpful for detecting E. histolytica.  designed to concentrate blood specimens suspected of containing low
- Coccidian parasites and microsporidia may also be recovered numbers of microfilariae
3. Cellophane Tape Preparation  1 mL of blood + 10 mL of 2% formalin
- specimen of choice for the detection of Enterobius vermicularis (pinworm) eggs. Adult  Mixed thoroughly and spun for 1 minute at 500 × g
female pinworms may also be seen.  Thick slides may be made, dried, and subsequently Giemsa-stained from the
- specimen should be collected in the morning before the patient washes or defecates. resulting sediment
- also used for recovery of Taenia spp. eggs  Buffy coat slides
 Buffy coat is a layer of white blood cells between the plasma and RBCs
 Microscopically examined for Leishmania and Trypanosoma
OTHER SPECIMENS AND LABORATORY TECHNIQUES:
 Collect oxalated or citrated blood, place it in a Wintrobe tube, and spinning it
1. Blood for 30 minutes at 100×g
- Parasites that may be recovered in blood include Leishmania donovani and  Cultures
Trypanosoma spp., Plasmodium and Babesia spp., and microfilariae  Novy-MacNeal-Nicolle (NNN) medium - yields favorable results for the
- Blood smears can be prepared from fresh whole blood without anticoagulant (fingertip recovery of Leishmania spp. and Trypanosoma cruzi
or earlobe) or from venipuncture collection with anticoagulant (EDTA).  Inoculated by the addition of a single drop of collected blood or ground
- If malaria is suspected, it is best to prepare smears within 1 hour of collection tissue.
 Thick smear  Penicillin is added to the medium if the specimen originates from a source
 for screening purposes; used when parasites are few in number or when thin that may contain bacteria.
smears are negative.  Negative cultures should be held for 1 month.
 Dehemoglobinized/laked
 Thin smear 2. CSF and Other Sterile Fluids
 recommended for species identification - To detect presence of morphologic forms of Naegleria fowleri and Acanthamoeba spp.,
 provide the best view of the malarial parasites in RBCs trypomastigote stages of Trypanosoma spp, Toxoplasma gondii and microsporidia and
 methanol-fixed Taenia solium cysticercus larvae and Echinococcus spp.
 Permanent Stains - If Naegleria or Acanthamoeba are suspected of being potential pathogens, the specimen
 two permanent stains commonly used: can be cultured on non-nutrient agar seeded with Escherichia coli.
 Wright’s stain
- contains the fixative and stain in one solution 3. Tissue and Biopsy Specimens
- yields only satisfactory results - recommended for recovery of intracellular organisms such as Leishmania spp. and T.
 Giemsa stain gondii.
- fixative and stain are separate - Other parasites that may be detected: free-living ameba, Trypanosoma spp., Trichinella
- considered the preferred stain because it allows for the detection of spiralis
parasite detail necessary for species identification. - Hepatic abscess material - specimen of choice for patients suspected of liver abscesses
caused by E. histolytica.

5
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

4. Sputum 10. Animal Inoculation and Xenodiagnosis

- typically collected and tested from patients suspected of being infected by the lung - Leishmania, Trypanosoma, and Toxoplasma infection may be tested by means of
fluke Paragonimus westermani. animal inoculation. Certain parasites have host specificity and require particular
- Strongyloides stercoralis hyperinfection patients will demonstrate motile larvae in animals. Mice, guinea pigs, and hamsters are used.
their sputum. - Xenodiagnosis – technique used for the diagnosis of Chagas’ disease.
- Other parasites may be found in sputum: microsporidia, E. histolytica, Entamoeba  An uninfected reduviid bug is allowed to take a blood meal from the patient and
gingivalis, Ascaris lumbricoides, and hookworm. the bug’s feces is then examined to observe for the presence of T. cruzi.
- Early-morning specimen is best and should be collected into a widemouthed
container with a screw cap lid.
- Saliva should not be mixed with the specimen. Sample may be examined directly via
PART III. AMEBAS
wet preps and/or concentrated using N-acetylcysteine or other appropriate agent.
- Equipped with pseudopods (false feet), which allows them move within their environment.
- Mode of transmission: ingestion of cyst from contaminated food or drink
5. Urine and Genital Secretions
- 2 morphologic forms:
- Urine is the specimen of choice for the detection of Schistosoma haematobium eggs
and may also yield Trichomonas vaginalis trophozoites Trophozoite Cyst
- Microfilariae can sometimes be found in the urine of patients with a heavy filarial Feeding stage Nonfeeding stage
infection. Undergoes replication via binary fission Does not undergo replication
- Saline wet preparations: method of choice for demonstrating the motile trophozoites. Form that feeds, multiplies, and possesses has a thick protective cell wall
pseudopods
6. Eye Specimens Susceptible to the environment outside the Can survive harsh environment
- Acanthamoeba keratitis is best diagnosed by the collection and examination of host
corneal scrapings. Not usually transmitted to humans. Usually transferred to humans via ingestion
- Small tissue samples be kept moist with sterile saline Encystation: troph → cyst; occurs in the Excystation: cyst → troph; occurs in the
- Calcofluor white stain: Acanthamoeba cysts stain apple green intestine ileocecal area

7. Mouth Scrapings and Nasal Discharge

LABORATORY DIAGNOSIS
- Mouth scrapings are the sample of choice for the detection of E. gingivalis and
- Trophozoites – soft, liquid, or loose consistency.
Trichomonas tenax
- Cysts – formed stool specimens
- Nasal discharge specimens are helpful for the recovery of parasites such as N. fowleri.
- Permanent smear of samples suspected of having amebas must be performed to confirm
parasite identification. It may, however, shrink amebic parasites, causing measurements
8. Skin snips
smaller than those typically seen in wet preparations.
- Useful in the detection of Onchocerca volvulus

9. Culture Methods
- Parasites that can be isolated with culture include E. histolytica, T. vaginalis,
Leishmania spp., T. cruzi, and T. gondii.

6
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Amebas Morphology
Cyst Trophozoite
# of nuclei Karyosome Peripheral Cytoplasm Cytoplasmic inclusions Motility Karyosome Peripheral Cytoplasm Cytoplasmic
chromatin chromatin inclusions
Entamoeba 1-4 Small and Fine, evenly Finely granular Cigar-shaped chromatoid Progressive, Small and Fine, evenly Finely granular Ingested RBCs
histolytica central distributed bars (rounded ends in unidirectional, central distributed
young cysts) finger-like
pseudopods
Entamoeba 1-8 Large, Unevenly Coarse and Splintered chromatoidal Nonprogressive, Large, Unevenly Coarse, Vacuoles containing
coli eccentric, distributed granulated bars; diffuse glycogen blunt Eccentric, distributed granulated bateria
irregular mass pseudopods irregular
Entamoeba 1-4 Small and Fine, evenly Finely granular Rounded end chromatoid Nonprogressive, Small, central Fine, evenly Finely granular Ingested bacteria
hartmanni central distributed bars in young cysts; finger-like distributed
Diffuse glycogen mass pseudopods
Entamoeba 1 Small and Fine, evenly Granular Splintered chromatoidal Progressive, Small and Fine, evenly Granular, Ingested bacteria,
poleckii central distributed bars in young cysts; unidirectional, central distributed vacuolated food particles
Diffuse glycogen mass in
young cysts
Endolimax 1-4 (4 most Large, Absent Granular, Chromatin granules; Sluggish, Large, Absent Granular, Bacteria
nana common) blotlike, vacuolated Nondescript small mass; nonprogressive, irregular, vacuolated
central Diffuse glycogen mass in blunt blotlike
young cysts pseudopods
Iodamoeba 1 (basket of Large, Absent Coarsely Well-defined glycogen Sluggish, Large, usually Absent Coarsely Bacteria, yeast cell,
butschlii flowers) eccentric granular and mass usually central granular, debris
achromatic vacuolated No chromatoid bars progressive refractive vacuolated
granules on achromatic
granules may
or may not
be present
Entamoeba ---NO CYST STAGE--- Centrally Fine, evenly Finely granular Leukocytes, Epithelial
gingivalis located distributed cells, Bacteria

7
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Significant Information
Entamoeba histolytica - The only known pathogenic intestinal amoeba
- Symptoms:
1. Asymptomatic carrier state
 Factors responsible to the carrier state
a. the parasite is a low-virulence strain
b. the inoculation into the host is low
c. the patient’s immune system is intact.
2. Symptomatic Intestinal Amebiasis
 amebic colitis - intestinal infection caused by the presence of amebas exhibiting symptoms.
 amebic dysentery - a condition characterized by blood and mucus in the stool
3. Symptomatic Extraintestinal Amebiasis
 Amebic liver abscess
 Venereal/vaginal amebiasis
 Lungs, pericardium, spleen, skin, brain
- TYI-S-33 - medium that supports E. histolytica in culture
Nice to know:
- Molecular analysis by PCR is the method of choice for discriminating between E. histolytica and non-pathogenic amebas.
- Entamoeba dispar, nonpathogenic ameba, is morphologically identical to E. histolytica.
Entamoeba poleckii - E. polecki has for a number of years been primarily considered a parasite of pigs and monkeys.
Iodamoeba butschlii - The term Iodamoeba was coined to describe an ameba that stains well with iodine. Unlike the other intestinal ameba, the nucleus of I. bütschlii does not undergo typical
division.
Entamoeba gingivalis - Specimen: mouth scrapings, particularly from the gingival area.
- E. gingivalis trophozoites have also been recovered in vaginal and cervical specimens from women who are using intrauterine devices (IUDs).
- Contracted via mouth-to-mouth (kissing) and droplet contamination, which may be transmitted through contaminated drinking utensils.

Infection caused Morphology Laboratory Diagnosis

Naegleria fowleri Primary Amebic Meningoencephalitis (PAM)
- invade the brain, causing rapid tissue
destruction.
- Kernig’s sign: diagnostic sign for meningitis,
patient is unable to fully straighten his or her
leg when the hip is flexed at 90 degrees
because of hamstring stiffness).
- Humans primarily contract this ameba by
swimming in contaminated water.
1. Ameboid Trophozoites 1. CSF – specimen of choice
- sluglike motility with blunt pseudopodia 2. Samples of tissues and nasal discharge
- cytoplasm – granular and often contain vacuoles - show a characteristic trailing effect when placed on agar
2. Flagellate Forms plates that have been previously inoculated with gram-
- pear-shaped negative bacilli.
- jerky or spinning motility with two whiplike flagella
3. Cysts
- exists only in the environment
- 1 nucleus, thick double wall
- cytoplasm – granular and often contains vacuoles

8
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Acanthamoeba Granulomatous Amebic Encephalitis (GAE) 1. Trophozoite 1. CSF

- MOT: nasal inhalation or aspiration, direct eye
invasion
- headaches, seizures, stiff neck, nausea, and
vomiting.
- granulomatous lesions of the brain are
characteristic and may contain both
Acanthamoeba trophozoites and cysts.
- sluggish, spinelike pseudopods (acanthapodia) - may be cultured on non-nutrient agar plates seeded with
- granular and vacuolated cytoplasm E. coli
2. Cysts 2. Corneal scrapings
- roundish with ragged edges - Calcofluor white: stain apple green
- double cell wall – smooth inner wall and wrinkled
outer wall
- cytoplasm – disorganized, granular, vacuolated

Acanthamoeba keratitis
- severe ocular pain and vision problems.

9
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

FLAGELLATES
- Flagella: whiplike structures responsible for the movement of flagellates
2. Chilomastix mesnili
- reside mainly in the small intestine, cecum, colon; Giardia intestinalis (duodenum)
- Undulating membrane - finlike structure connected to the outer edge of some flagellates Trophozoite Shape Pear-shaped
- Axostyle - a rodlike support structure Motility Stiff, rotary, directional
- Nuclear characteristics, such as number of nuclei present and the presence and Nuclei One with small central or
positioning of the nuclear structures, are helpful in differentiating the flagellates. eccentric karyosome
No peripheral chromatin
1. Giardia intestinalis Flagella Four:
Infection 1. Giardiasis (Traveler’s diarrhea) Three extending from anterior end
caused - mild diarrhea, abdominal cramps, anorexia, and flatulence to tenderness of One extending posteriorly from cytostome region
the epigastric region, steatorrhea, and malabsorption syndrome Other Prominent cytostome extending 1/3 to 1/2 body
2. Asymptomatic carrier state structures length
- usually cases are asymptomatic Spiral groove
MOT Ingestion of contaminated water or food Cyst Shape Lemon-shaped, with a clear hyaline knob
Person-to-person contact (oral-anal) extending from the anterior end
Trophozoite Shape Pear-shaped, teardrop Nuclei One, with large central karyosome
Motility Falling leaf No peripheral chromatin
Appearance Bilaterally symmetrical Other Well-defined cytostome located on one side of the
Nuclei Two ovoid-shaped, each with a large karyosome structures nucleus
No peripheral chromatin
Flagella Four pairs, origination of each: 3. Dientamoeba fragilis
One pair, anterior end Trophozoite Shape Irregularly round
One pair, posterior end Motility Progressive, broad hyaline pseudopodia
Two pair, central, extending laterally Nuclei Two, each consisting of massed clumps of four to
Other structures Two median bodies, two axonemes, sucking disk eight chromatin granules
Cyst Shape Ovoid No peripheral chromatin
Nuclei Immature cyst, 2; Mature cyst, 4 Cytoplasm Bacteria-filled vacuoles
Central karyosomes, no peripheral chromatin NO CYST STAGE
Cytoplasm structures Retracted from cell wall Significant - can be transmitted via the eggs of helminth parasites such as
Other structures Median bodies: 2 in immature cyst, 4 in fully mature Information Enterobius vermicularis and Ascaris lumbricoides
cyst; interior flagellar structures
Significant - Stool: specimen of choice
Information - Enterotest: string test
- Cysts: can survive in the environment for up to 3 months; resistant to
chlorination
- G. intestinalis, along with Trichomonas vaginalis, are known to be carriers of
dsRNA viruses.
- IgA deficient patients: susceptible to reoccurring infections

10
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

4. Trichomonas spp

Specimen of choice Trophozoite

Shape Motility Nuclei Flagella Undulating
Membrane
Trichomonas Stool Pear-shaped Nervous jerky One, with small central 3-5 anterior; 1 posterior Full body length
hominis karyosome
Trichomonas Urine, Ovoid, round or Rapid, jerky One, ovoid, nondescript 3-5 extending anteriorly; 1 extending ½ of body length
vaginalis Vaginal discharge pear-shaped posteriorly
Urethral discharge
Prostatic secretions
Trichomonas tenax Mouth scrapings Oval, pear-shaped Jerky One, ovoid nucleus; consists 5 total, all originating anteriorly: 2/3 of body length
of vesicular region filled 4 extend anteriorly
with chromatin granules 1 extends posteriorly

Trichomonas vaginalis HEMOFLAGELLATES

- MOT: sexual intercourse, migration through birth canal
Morphology and Life Cycle
Clinical symptoms: Appearance Nucleus Location of Undulating Flagella Specimen
1. Asymptomatic kinetoplast membrane
- Mostly men Amastigote Round to 1, usually Near None None Tissue,
2. Persistent urethritis oval off center nucleus muscle, CSF
- Condition that symptomatic men experience (within
- Enlarged tender prostate, dysuria, nocturia, and epididymitis macrophages)
- Thin, white urethral discharge Promastigote Long and 1, located Anterior None Peripheral
3. Persistent vaginitis slender near or in end Extends blood
- Foul smelling, greenish-yellow liquid vaginal discharge center from
Epimastigote Long and 1, located Anterior to Half of anterior Blood (only if
- Most symptoms: Urethral involvement, dysuria, and increased frequency of urination
slightly in nucleus body end collected
- Burning, itching, and chafing may also be present. Red punctate lesions
wider than posterior immediately
4. Infant infections promastigote end after
- Trophozoites migrate from mother to infant through the birth canal or during vaginal transmission)
delivery Trypomastigote C, S, U shape, 1, located Posterior Entire body Found in
- Respiratory infection and conjunctivitis long and anterior to end vector
slender the
Lab Diagnosis:
kinetoplast
1. Wet preparation
2. Culture
3. Serology

11
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Leishmania spp. Trypanosoma East African - Fever, myalgia, rigors - Specimen: blood, CSF
- MOT: bite of infected vector brucei Sleeping - No lymphadenopathy - Microscopic exam,
- Vector: Sandfly (Lutzomiya, Phlebotomus, Psychodogus) rhodesiense Sickness, - May lead to IgM detection in serum
- Infective stage: Promastigote Rhodesian glomerulonephritis and or CSF
- Diagnostic stage: Amastigote trypanosomiasis myocarditis

Associated Disease Symptoms Specimen

Leishmania Mucocutaneous Leishmaniasis Large ulcers in the Biopsy of ulcer Trypanosoma cruzi
braziliensis Espundia, forest yaws, pian bois, oral or nasal mucosa - Causative agent of Chaga’s disease
uta - MOT: bite of infected vector, blood transfusions, sexual intercourse, transplacental
Leishmania Visceral Leishmaniasis Hepatosplenomegaly, Blood, bone transmission, and entry through the mucous membranes
donovani Kala-azar, dumdum fever abdominal illness, marrow, - Vector: Reduviid bug, kissing bug, conenose bug, triatomid bug (Panstrongylus
darkening of the skin lymph node megistus)
aspirate, - Infective stage: Trypomastigote
biopsy
- Diagnostic stage: Trypomastigote/Amastigote
Leishmania New World Leishmaniasis Affects ear and Lesion biopsy
mexicana Chiclero ulcer, bay sore surrounding Associated Symptoms Lab Diagnosis
cartilage Disease
Leishmania Old World Leishmaniasis One or more ulcers Fluid Trypanosoma American - chagoma – site of - Specimen: blood
tropica Oriental sore, Delhi/Baghdad boil, containing pus underneath cruzi typanosomiasis infection - Microscopic exam,
dry or urban cutaneous Thick plaque of skin ulcer bed - Romana’s sign – complement fixation,
leishmaniasis with multiple lesions conjunctivitis and DAT, IIF, PCR, ELISA
or nodules unilateral edema of the - Xenodiagnosis
eyelids
Trypanosoma brucei gambiense & Trypanosoma brucei rhodesiense - Chronic phase –
- Causative agent of nagana and sleeping sickness megacolon,
- MOT: bite of infected vector megaesophagus,
- Vector: Tsetse fly cardiomegaly
- Infective stage: Trypomastigote
- Diagnostic stage: Trypomastigote Trypanosoma rangeli
Associated Symptoms Lab Diagnosis - No known disease association
Disease - Vector: Reduviid bug (Rhodium prolixus)
Trypanosoma West African - Chancre - Specimen: blood, - Specimen: blood
brucei Sleeping - Winterbottom’s sign lymph node aspirate, - Lab diagnosis: xenodiagnoses and serology
gambiense Sickness, (lymphadenopathy) CSF
Gambian - Kernadel’s sign (delayed - Microscopic exam,
trypanosomiasis sensation to pain) IgM detection in serum
- Somnolence (excessive or CSF
sleepiness)

12
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

SPOROZOA: Plasmodium and Babesia

- Phylum Apicomplexa 2. Sporogony/Sexual Cycle
- No means of motility - The gametocytes, microgametocytes and macrogametocytes, are ingested by an
Anopheles mosquito during a blood meal.
Plasmodium spp. - While in the mosquito’s stomach, the microgametes penetrate the macrogametes
- P. falciparum: most common in the Philippines and tropics generating zygotes.
- P. vivax: most common worldwide - The zygotes in turn become ookinetes, which invade the midgut wall of the mosquito
- Mixed infection with P. vivax and P. falciparum where they develop into oocysts.
- Hypnozoites: P. vivax and P. ovale - The oocysts grow, rupture, and release sporozoites, which make their way to the
- Vector: Anopheles mosquito’s salivary glands.
- MOT: Bite of infected vector, blood transfusion, parenteral transmission - Inoculation of the sporozoites into a new human host perpetuates the malaria life cycle.
Clinical Symptoms
Life Cycle
- Paroxysms: recurrent fever, chills, and sweating due to synchronized rupture of RBCs
1. Schizogony/Asexual Cycle
releasing merozoites
A. Exoerythrocytic stage
o Every 36 hours: Malignant Tertian Malaria (P. falciparum)
- Anopheles infects man with sporozoites
o Every 48 hours: Benign Tertian Malaria (P. ovale and P. vivax)
- Sporozoites are carried to the liver where they will develop into merozoites
o Every 72 hours: Quartan malaria (P. malariae)
- Infected hepatocytes rupture and introduce the merozoites into the bloodstream
- Anemia, splenomegaly, joint pain, ischemia
B. Erythrocytic stage - Severe complications:
- Merozoites infect red blood cells becoming ring stage trophozoites that mature into o Cerebral malaria – RBCs, parasites, and pigment block brain vessels
schizonts, which rupture releasing more merozoites o Blackwater fever – sudden massive intravascular hemolysis resulting to
- Merozoites invade new RBCs and multiply hemoglobinuria
- Some merozoites differentiate into gametocyte

Morphological Features of Malarial Parasites

P. vivax P. ovale P. malariae P. falciparum

Disease Benign tertian Benign tertian Quartan Malignant tertian
Trophozoite Amoeboid Amoeboid, but not as evident as P. vivax Band/bar Ring
Merozoites 12-24 (16) 8, rosettes 6-12, fruit pie/rosette (central brown pigment) 8-36 (24)
Microgamete Round to oval Round to oval Round to oval Crescent, sausage, banana
Macrogamete Round to oval Round to oval Round to oval Crescent, sausage, banana
Infected RBCS Young, enlarged, pale Young, enlarged; large, serrated, fimbriated Mature, normal/smaller Young/mature; normal
Paroxysms Every 48 hours Every 48 hours Every 72 hours Every 36 hours
Stipplings Schuffner’s dots James’ dots Ziemann’s dots Maurer/Cuneiform/Stephen Cristopher’s dots
Stages present in all all Ring forms; growing and mature trophozoites Ring form and gametocytes
peripheral blood and schizonts

13
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Plasmodium knowlesi Resistance to Malaria

- Fifth human malarial parasite, normal a parasite of long-tailed macaques 1. Duffy negative
- Described in humans in the PH and SEA - Resistance to P. vivax and P. knowlesi
- Microscopically indistinguishable from P. malariae (older stages), and differentiation is only 2. M-N-
achieved through PCR (cross-reaction with P. vivax interferes PCR test) - Resistance to P. falciparum
- Early ring stages resemble those of P. falciparum 3. G6PD deficiency
4. Sickle cell anemia

Lab Diagnosis Babesia

1. Examination of blood film (Giemsa-stained) - Causative agent of Texas cattle fever and red water fever
- Gold standard - Life cycle is similar to Plasmodium spp. except no exo-erythrocytic stage
- Blood collected using EDTA: preferred - IS: sporozoites
- Finger-stick blood: recommended when volume of blood required is minimal - DS: trophozoites, merozoites
- 200 – 300 OIO fields should be examined - MOT: bite of infected vector (Ixodes ticks), blood transfusion
A. Thick smear - Mimics P. falciparum ring forms
- Screening - Merozoites: Maltese cross appearance
- Dehemoglobinized: dipped in water
B. Thin smear
- Used to differentiate Plasmodium species
- Fixed with methanol CILIATES
Balantidium coli
2. Quantitative Buffy Coat - Only human pathogen
- Acridine orange: (+) bright green/yellow fluorescence - Largest parasitic protozoan
- MOT: Ingestion of infected cysts
3. Immunologic - Natural host: pigs
A. Optimal assay - IS: cyst
- Test for parasitic LDH - DS: cyst, trophozoites
- Produced by all 4 species of Plasmodium
- 100-200 parasites/uL Morphology:
B. Malaquick 1. Trophozoite: (“little bag”)
- Histidine-Rich Protein 2 (HRP2) - Macronucleus: kidney-bean, vegetative function
- P. falciparum - Micronucleus: small, spherical, reproductive
4. PCR - Anterior end has cytostome; layer of cilia around the organism
- Can detect 5 to 10 parasites per uL - Motility: rapid, boring, rotary

2. Cyst:
- Double cyst wall enclosing the cilia
- Contains macronucleus and micronucleus (may not be observable)

14
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Clinical symptoms: Blastocystis hominis

- Balantidiasis: abdominal discomfort, dysentery - 4 forms: cyst, central vacuole/vacuolated (most common), granular (cultures), amoeboid,
- Can invade tissues causing submucosal lesions and hemorrhage - Flies and cockroaches can harbor this parasite
-
COCCIDIANS Cyclospora cayatenensis
Isospora belli - Affects immunocompromised and immunocompetent patients
- Only coccidian with no intermediate host - Longer diarrhea
- Definitive host: man - Associated with biliary disease
- IS: sporulated oocyst - Associated with outbreaks linked to contaminated water and fresh produce
- DS: oocyst - Oocysts sporulate best at room temp (10 to 12 days)
- MOT: ingestion of sporulated oocyst in contaminated food/water, oral-anal sexual contact - Auto-florescence: blue-green
- Modified acid-fast stain: wrinkled cellophane with bubbly appearance
Laboratory diagnosis:
- Specimen: stool, duodenal content, intestinal biopsy Microsporidia
- Sheather’s sugar flotation
- May causes enteritis, keratoconjunctivitis, myositis
- Modified acid-fast
- Infections mostly occurs in immunocompromised patients
- Charcot-Leyden crystals in the stool
- Acid-fast and Periodic Acid-Schiff: positive
- Electron microscopy to identify species
Sporoblast Sporocyst
Immature oocysts Mature, infective oocyst
Shed in stool Found in environment
Cysts surrounded by smooth, colorless, two-layered cell wall Toxoplasma gondii
- DH: cats (sporogony/sexual stage occurs) IH: man (schizogony/asexual stage occurs)
Cryptosporidium parvum - IS: oocyst
- Immunocompromised patients; causes diarrhea in AIDS patients - MOT: ingestion of infective oocyst, transplacental transmission, blood transfusion
- Agent responsible for neonatal diarrhea in calves and lambs
Morphology
- Autoinfection may occur
Oocyst Tachyzoite Bradyzoites
- G. lamblia, D. fragilis, and Cryptosporidium have been implicated in nursery school
- Consists two sporocysts, - Actively dividing form - Inactive/slow-growing form
outbreaks of diarrhea
each with four sporozoites - Found in cells of the IH and - Found within tissue cysts
- G. lamblia and Cryptosporidium have been implicated in waterborne outbreaks of diarrhea
- Bordered by a clear, in epithelial cells of DH and multiply very slowly
colorless, two-layered cell - Crescent-shaped - Appears in clusters
Laboratory diagnosis: wall - has a single centrally located - PAS-positive
- Specimen of choice: stool (ova), intestinal biopsy (schizonts, gametocytes) - Resistant to disinfectants, nucleus, surrounded by a cell
- Sheather’s sugar flotation freezing, drying (killed by membrane
- Modified acid-fast, Enterotest, ELISA, IFA heating at 70C for 10 mins

15
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Laboratory Diagnosis Classification According to the Presence of Phasmids/Caudal Chemoreceptors:

- Specimen: blood 1. Aphasmid (without caudal chemoreceptors) (Adenophorea): Trichuris, Trichinella,
1. ELISA Capillaria
- Recommended for IgM determination in congenital infection 2. Phasmid (with caudal chemoreceptors) (Secernentia): the rest of nematodes
2. Sabin-Feldman Dye test
- Methylene blue staining of tachyzoites inhibited by prior addition of patient serum Classification based on the Habitat of the Adult Worms
containing antibodies to Toxoplasma (Inhibition mechanism) Small intestine Extraintestinal Nematode
- (+) trophozoites did not absorb the dye Capillaria philippinensis Lymph node and Lymphatic vessels
- (-) trophozoites absorbed the dye Ascaris lumbricoided Filarial worms
Strongyloides stercoralis - Wuchereria
Pneumocystic jivorecii Hookworms - Brugia
Trichinella spiralis (adult)
- Lung infection in immunocompromised patients Eyes and meninges
- Stain: Grocott methenamine silver stain Large intestine Angiostrongylus/Parastrongylus cantonensis
Enterobius vermicularis Encysted in the host muscle
Trichuris trichiura Trichinella spiralis (larva)

NEMATODES
General characteristics: Ascaris lumbricoides
1. Adult nematode is elongated, cylindrical worm primarily bilaterally symmetrical - Giant intestinal roundworm; most common intestinal nematode of man
2. Anterior end may be equipped with hooks, teeth, plates, and papillae for purpose of abrasion, - Vector: Flies and cockroach
attachment and sensory response - Member of Soil Transmitted Helminths (STH): HATS
3. Supporting body wall: (1) outer hyaline, noncellular cuticle, (2) subcuticular epithelium, (3) - Member of Unholy Trinity: HAT
layer of muscle cells - Member of parasites having Lung-Heart Migration: ASH
4. Alimentary tract is simple tube extending from mouth to anus. NO circulatory system
5. Complete digestive and reproductive system Habitat Small intestine
6. Life cycle: egg stage > larval stage > adult stage Final host Man
7. They are provided with separate sexes (Dioecious) although some may be parthenogenetic Diagnostic stage Ova (fertilized/unfertilized)
(Strongyloides stercoralis) Infective stage Embryonated egg
Female worms produce eggs: MOT ingestion of embryonated eggs
a. Oviparous – lay immature/unembryonated/unsegmented eggs (Ascaris)
b. Oviviparous/Ovoviviparous – lay mature/embryonated eggs (Enterobius) Morpholology:
c. Viviparous/Larviparous – lay larva, not eggs (Trichinella) 1. Adult
8. There are sensory organs (chemoreceptors) in the anterior and posterior ends of the worm - white, cream, pinkish yellow when fresh
called amphids (cephalic chemoreceptors) and phasmids (caudal chemoreceptors) - trilobate lips (triangular buccal cavity
Female Male
Color Creamy white, pink tint Creamy white, pink tint
Other features Pencil lead thickness Prominent incurved tail

16
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Enterobius vermicularis (Oxyuris vermicularis)

2. Eggs - Common name: Pinworm, Social worm, Society worm
- Becomes infective 2-6 weeks after deposition - Has familial or group tendency
- Survive harsh environment - Most common helminth to infect man worldwide
Layer Unfertilized Fertilized Corticated Fertilized Decorticated Habitat Large intestine
Inner No vitelline membrane vitelline membrane vitelline membrane Final host Man
Middle Glycogen Glycogen Glycogen Diagnostic stage Embryonated egg
Outer Refractile lecithin Albuminous/mamillary coat Chitin Infective stage Embryonated egg
granules MOT Ingestion, inhalation, autoinfection

Life cycle: Morphology:

1. Adult lives in small intestines 1. Adult
2. Undeveloped eggs in feces embryonate in warm moist soil - Small, whitish, brownish
3. Embryonated eggs ingested by man - Anterior end: lateral wings or cephalic alae
4. Larva hatch in small intestine and begin larval migration - Posterior: bulb-like esophagus
- Male: dies after copulation
Symptoms and Pathology: - Female: goes to perianal region to deposit eggs; has pointed tail resembling pinhead
1. Vomiting and abdominal pain 2. Eggs
2. Pneumonia, cough, fever, eosinophilia during larval migration - D-shaped (flattened on one side), embryonated
3. Possible intestinal perforation - Double-layered: outer albuminous layer, inner lipoidal layer
Adult: may migrate to extraintestinal site (hepatic ascariasis, kidney, etc.)
Larva: Heart-Lung Migration Life cycle:
- May resemble Loeffler’s syndrome / Ascaris pneumonitis 1. Adult in colon
- Eosinophilia; Charcot-Leyden crystals 2. Gravid female migrates to perianal region to deposit eggs
3. Eggs ingested, hatch in small intestine, releasing larvae and develop into adult worms
Laboratory Diagnosis:
- Specimen: Stool Symptoms and Pathology:
1. Pruritis ani/Nocturnal Pruritis ani
Additional notes: 2. Hemorrhagic colitis
1. Toxocara cati – cats 3. Extraintestinal enterobiasis: vagina, uterus, fallopian tube
2. Toxocara canis – dogs; can be ingested by man and cause Visceral Larva Migrans (Ocular 4. Autoinfection
Larva Migrans)
Laboratory:
1. Perianal swab
2. Scotch tape swab/cellulose tape swab
3. Stool – 5% survival

17
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Trichuris trichiura Morphology:

- Common name: Whipworm 1. Adult
- Part of Unholy Three - Delicate tiny worms
Habitat Large intestine - Male with chitinized spicule
Final host Man - Females usually have eggs in utero
Diagnostic stage Ova found in stool 2. Egg
Infective stage Embryonated egg - Guitar, peanut-shaped
MOT Ingestion of embryonated egg - With bipolar mucus plugs, striated and smaller than T. trichiura egg
Morphology: - Difference from T. trichiura egg:
1. Adult  smaller and striated
- Flesh-colored or pinkish gray, slender or attenuated in the interior resembling a whip  mucus plugs are not as protruded
2. Eggs Typical egg Atypical egg
- “Japanese lantern”; barrel/football-shaped with bipolar mucus plug - immature eggs - segmented egg
- Prominent hyaline polar plug - unembryonated eggs - embryonated egg
Life Cycle: - will go to stool - hatch inside the small intestine
1. Adult in the colon - will go to fresh water where - responsible for autoinfection
2. Undeveloped eggs in feces, embryonate in warm most soil embryonation process occurs
3. Embryonated egg ingested by man - eaten by fish
4. Larvae hatch in small intestine, penetrate and develop in villi, return to lumen and migrate to
cecum, mature to adult Symptoms and Pathology:
1. Malabsorption: steatorrhea
Symptoms and Pathology:
2. Borborygmi: abdominal gurgling sound
- Trichuriasis, trichocephalis, whipworm infection
3. Abdominal pain and diarrhea
1. Loody or mucoid diarrhea
2. Weight loss, abdominal pain
3. Rectal prolapse (heavy infection)
Hookworms
- 2nd most common helminthic infection reported in humans
Capillaria philippinensis
- Common name: Pudoc worm: Pudoc disease/Mystery disease Ancylostoma duodenale Old World Hookworm 2 pairs of teeth
- Close relative of Trichuris trichiura Necator americanus New World Hookworm Semilunar cutting plates
Ancylostoma braziliense Cat Hookworm 2 pairs of teeth
Habitat Small intestine Ancylostoma caninum Dog Hookworm 3 pairs of teeth
Final host Man
Natural host Migratory birds (Final host in the environment) Habitat Small intestine Infective stage L3/Filariform Larva
Intermediate host Fresh water/Brackish water fish (Ipori, Birot, Bagsang) Final host Man MOT Skin penetration
Diagnostic stage Larva and/or ova found in stool Diagnostic stage Egg/Larva
Infective stage Larval stages found in the infected fish
MOT Ingestion of of raw/undercooked fish bearing infective larva

18
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Morphology: 3. Adult
1. Adult - Tissue damage at the site of attachment
Necator americanus Ancylostoma duodenale - Enteritis and pain: blood loss and anemia
Shape S-shaped C-shaped - Iron deficiency anemia
Buccal cavity Semilunar cutting plates 2 pairs of teeth - Acutegastrointestinal phase demonstrate increased eosinophilia
Copulatory bursa Bipartite (2 digits) Tripartite (3 digits)
Barbed/bristle-like Simple, not barbed Ancylostoma Eos peak: approx. 1 month Associated with vertical transmission and
duodenale congenital infections
2. Eggs Necator Eos peak: approx. 2 months Skin-associated symptoms
- Same for all species (indistinguishable) americanus
- Ovoidal, thin-shelled, and colorless
- “Morula ball” formation – 8 cell stage; germ cell in fragmentation Laboratory Diagnosis
1. Stool Exam
3. Larva 2. Haradamori Culture/Filter Paper Culture Technique
L1 Rhabditiform Larva L3 Filariform Larva 3. Baermann Funnel Technique
1st stage larva 3rd stage larva
Open mouth: feeding stage Closed mouth: nonfeeding stage
Short and stout Long and slender
Long buccal cavity Sheathed, pointed tail Strongyloides stercoralis
Short/small genital primordium - Common name: Threadworm
- Smallest nematode infecting man
Life cycle: - Facultative nematode
1. Adults live in small intestine
2. Egg in feces, develops rapidly to rhabditiform larva and hatches in soil Habitat Small intestine
3. After molting twice, become filariform larva Final host Man
4. Filariform larva penetrates skin of man, begins larval migration Diagnostic stage Rhabditiform (egg signifies heavy infection)
Infective stage L3/Filariform Larva
MOT Skin penetration. Internal autoinfection
Symptoms and Pathology
1. Larval Penetration
Morphology:
a. Human Hookworm
1. Larva
 Allergic reaction: Ground itch/Dew itch/Water sore/Mazza Mora
Hookworm Threadworm
 Severe itching at the site of larval penetration of skin
L1 Rhabditiform larva
b. Animal Hookworm
Buccal cavity Long Short
 Creeping eruption or Cutaneous Larva Migrans Genital primordium Small/inconspicuous Prominent/conspicuous
L3 Filariform Larva
2. Larval Migration Sheath Sheathed Unsheathed
- Heart-Lung Migration: Wakana disease Tail Pointed Notched
 Pulmonary lesions and alveolar hemorrhage. Bloody sputum

19
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

2. Eggs Trichinella spiralis

- Chinese lantern appearance - Common name: Trichina Worm/Muscle Worm
- Rarely found; diagnosed as hookworm egg - All developmental stages occur within a single host
- Slightly smaller than hookworm egg; well-developed contained in the egg
Habitat Adult: Small intestine
Larva: striated skeletal muscle
Symptoms and Pathology:
Final host Pigs and other mammals; Man (accidental/dead end host)
- Strongyloidiasis, Cochin China Diarrhea, Vietnamese Time bomb
Diagnostic stage Encysted larva (muscle biopsy)
Infective stage Encysted larva
1. Larval Penetration
MOT Ingestion of undercooked/raw meat with encysted larva
- Allergic reaction at the site of penetration
- Larva currens: tracks under the skin from worm migration Morphology:
2. Larval Migration 1. Adult
- Heart-Lung migration: resembles bronchopneumonia - Male: conical papillae
3. Adult - Female: club-shaped uterus
- Cochin China Diarrhea, Vietnamese Time bomb 2. Larva
- Honeycomb appearance of the intestinal mucosa - Nurse cell, a striated muscle cell, surrounds the coiled larva
4. Autoinfection
- Infective larva penetrates intestinal mucosa Life Cycle Notes:
1. Man ingests undercooked meat containing larva (from pig, bear)
Laboratory Diagnosis: 2. Larva digested out of muscle in intestine, maturing to adults
1. Stool Exam 3. Viviparous female releases larvae penetrating intestinal mucosa and disseminate into
- DS: Rhabditiform larvae bloodstream
2. Baermann Funnel Technique 4. Larvae encyst in striated muscle
3. Harada Mori Filter Paper Culture Technique
- Recommended culture method Symptoms and Pathology:
- Trichinosis, Trichiniasis, Trichinellosis; Great imitator
1. Larva
Strongyloides fuelleborni - Larval migration causes fever, facial edema, eosinophilia
- Muscle pain at site of encystation
- Swollen belly syndrome
- Difficult in swallowing and breathing
- A primate parasite isolated from humans in Africa
2. Adult
- Diarrhea and abdominal pain

Laboratory:
1. Muscle biopsy
2. Immunotests
- Bentonite Flocculation Test, Bachmann Intradermal Test, Beck’s Xenodiagnosis

20
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Filarial worms 5. Larva migrates and develop to adults in tissue of man

Wuchereria bancrofti Brugia malayi
Movement Graceful/smooth Kinky/stiff Lab diagnosis:
Nuclei Not overlapping overlapping 1. Fluid examination
Sheath Sheathed Sheathed - Specimen: blood, urine, hydrocele fluid, chyle
Terminal Nuclei No terminal nuclei Bulge around 2 terminal nuclei - Stained with Giemsa
2. Knott’s concentration technique
Habitat Adult: Lymphatic system - 1mL of blood + 10mL in 2% solution, centrifuged at 500x g for 1 minute
Microfilaria: Blood - Slide is stained with Giemsa
Loa loa, Onchocerca volvulus: subcutaneous tissue 3. Nucleopore Filtration
Final host Man - Blood is passed through a polycarbonate filter (2um pore)
Intermediate host Mosquito/arthropod - Distilled water is passed through the filter, lysing the red cells
Diagnostic stage Microfilaria - Filter is stained with Giemsa
Infective stage IS to IH: microfilaria 4. Multiplex PCR
IS to FH: Filariform larva - To differentiate W. bancrofti from B. malayi
MOT Bite of infected arthropod
Symptoms and Pathology:
Life Cycle:
1. Larva: Tropical Pulmonary Eosinophilia
1. Adult in lymphatics, subcutaneous tissue; microfilaria in blood
2. Adult: Lymphatic Filariasis
2. Microfilaria ingested by intermediate host
- Elephantiasis: wrinkled, dry appearance of extremities
3. Microfilaria develops into infective stage in arthropod
- Hydrocele: inflammation of the scrotum
4. Infective larva enters skin at bite site
- Chyluria: milky white urine due to lymphatic rupture and fluid entering the urine

Habitat Vector Specimen Microfilaria Periodicity

Wuchereria bancrofti Lower Aedes, Anopheles, Blood Sheathed Nuclei absent in tail Nocturnal (9pm
- Bancroft’s Filarial worm lymphatics Culex to 4am)
- Hydrocoele
- Elephantiasis
Brugia malayi Upper Mansonia, Anopheles Blood Sheathed Tail with 2 separate nuclei Subperiodic,
- Malayan filarial worm lymphatics nocturnal

Loa loa Subcutaneous Chrysops, Tabanid fly, Blood Sheathed Nuclei continuous up to the tip of the tail Diurnal
- Eyeworm tissue Deer fly, Mango fly 10:15am –
- Calabar swelling, fugitive 2:15pm
swelling

21
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Onchocerva volvulus Subcutaneous Simulium, Blackfly Skin snips Unsheathed Nuclei absent in tail Nonperiodic
- Blinding worm, tissue
Convoluted Filaria
- River blindness, hanging
groin
- Leopard skin
Mansonella ozzardi Body cavities Culicoides Blood Unsheathed Nuclei absent in tail Nonperiodic

Mansonella perstans Body cavities Culicoides Blood Unsheathed Nuclei up to the tip of the tail Nonperiodic
- Joint and bone pain,
enlargement of liver

Additional notes:
- Tropical Eosinophilia/Occult Filariasis – condition where persons who reside in areas Treatment Removal of worm by immersing the affected part in
where both B. malayi and W. bancrofti are endemic. cold water. Worms are then slowly retracted using
- W. bancrofti, Brugia spp., and Onchocerca volvulus harbor an endosymbiotic alpha- a stick.
proteobacterium, Wolbachia sp. Wolbachia is an obligate intracellular organism. The
parasites require the endosymbiont for larval development, viability, and fertility Life Cycle:
1. Man ingests copepods infected with 3rd stage larva
Dirofilaria immitis 2. Larva matures into adult worms, penetrate the intestinal wall, and proceed to connective
- Dog Heartworm tissue or body cavities
- Common filarial parasite of dogs 3. Gravid female worms migrate into the subcutaneous tissue, laying first-stage larvae.
- Coin lesions: solitary, peripheral nodules in the lung 4. An infected ulcer results at the site of the larvae deposit.

Dracunculus medinensis
- Common name: Guinea worm, Fiery Serpent of the Israelites, Medina worm, Serpent
worm
- Longest nematode of man
Habitat Subcutaneous tissue
Intermediate host Aquatic crustacean (copepods)
Diagnostic stage Rhabditiform larva (1st stage)
Infective stage 3rd stage larva
MOT Ingestion of copepods

22
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Parastrongylus (Angiostrongylus) cantonensis TREMATODES

- Common name: Rat lungworm General characteristics:
1. All flukes appear flat and leaf-like except Schistosomes (elongated and cylindrical)
Intermediate host Mollusks/snail 2. All are hermaphroditic except Schistosomes (dioecious/separate sexes)
Natural host Rats 3. All eggs are operculated except Schistosomes (nonoperculated)
MOT Ingestion of intermediate host containing larva 4. Life cycle of trematodes: Egg → Larva → Adult
Adult morphology Barber’s pole appearance; looping of whitish
uterus to the red digestive tract Larva: Miracidium → Sporocyst → Redia → Cercaria → Metacercaria
Symptoms and Cerebral angiostronygliasis ***Except Schistosomes (no metacercaria)
Pathology - Eosinophilia and symptoms of meningitis 5. Infective stage to the FH: Metacercaria except Schistosomes (cercaria)
- CSF contains ↑ WBCs and eosinophils
6. MOT: Ingestion except for Schistosomes (skin penetration)
7. Require 2 intermediate host except for Schistosomes (only 1 IH: snail)
Parastrongylus costaricensis
- Found primarily in cotton rat and black rat  1st IH: snail (Cercaria)
- Abdominal angiostrongyliasis (abdominal pain similar to appendicitis)  2nd IH: fish, crab, plant, snail, ant (Metacercaria)
- Larva create inflammatory lesions in the wall of the bowel, resulting in tissue  Fish – Clonorchis sinensis, Heterophyes heterophyes, Opistorchis felineus
inflammation, necrosis, vomiting and diarrhea  Crab – Paragonimus westermani
- MOT: ingestion of salad contaminated with infected slugs or snails  Plant – Fasciola hepatica, Fasciola gigantica, Fasciolopsis buski
 Snail – Echinostoma ilocanum
Anisakis spp.  Ant – Dicrocoelium dendriticum, Eurytema pancreaticum
- Common name: Herring’s worm 8. Adults attach themselves to the host by means of 2 suckers: oral sucker, ventral sucker
- Disease: Herring disease (acetabulum) except for Heterophyes heterophyes (3 suckers: genital sucker/gonotyle)
Definitive host Whales/Dolphins 9. Egg
1st IH Copepods  Mature when laid
2nd IH Smaller fishes  S. schistosoma
3rd IH Larger fishes
 H. heterophyes
MOT Ingestion of raw fish infected with larva
 O. volvulus
Symptoms & Granuloma around migrating larva
Pathology  C. clonorchis
 Immature when laid
 Fasciola
 Fasciolopsis
 E. ilocanum
 P. westermani
10. Possess alimentary canal without anus (incomplete) and complex reproductive structures
(testes, ovary, uterus)
11. Treatment: Praziquantel

23
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

A. LIVER FLUKES 4. Opistorchis felineus

1. Fasciola hepatica Common name Cat Liver Fluke/Siberia Liver Fluke
Common name Sheep Liver Fluke/Temperate Liver Fluke Habitat Liver/Bile passages of the liver
Habitat Biliary duct/Liver Final host Cats
Final host Sheep 1st IH Snail
1st IH Snail 2nd IH Fish
2nd IH Plant/Vegetation Egg Operculated, mature
Egg Large, operculated, unembryonated, yellowish brown ***Opistorchis viverrini – SouthEast Asian Liver Fluke; anti-P1
egg resembling F. gigantica and F. buski eggs
Hen’s egg shape 5. Dicrocoelium dendriticum/lanceolata
Adult Cephalic cone, well-developed shoulder Common name Lanceolate/Lancet Fluke
Habitat Bile passages of the liver
2. Fasciola gigantica Final host Cattle/Sheep
Accidental host Man
Common name Giant Liver Fluke/Tropical Liver Fluke 1st IH Snail
Habitat Biliary passages of liver 2nd IH Ants
Final host Cattle Egg Brownish, thick-walled, operculated, embryonated
1st IH Snail Adult Lancet-shaped
2nd IH Plant/Vegetation
Egg Fasciola egg B. INTESTINAL FLUKES
Adult Less distinct cephalic cone, less developed shoulder
1. Fasciolopsis buski

3. Clonorchis sinensis Common name Giant Intestinal Fluke

Habitat Small intestine
Common name Chinese Liver Fluke/Oriental Liver Fluke Final host Man
Habitat Bile duct/Gall bladder Reservoir host Pig, dogs, rabbits
Final host Man 1st IH Snail
1st IH Snail 2nd IH Plants/Vegetation
2nd IH Fish Egg Indistinguishable from Fasciola eggs
Egg Old-fashioned electric light bulb; pitcher-like
Small abopercular knob 2. Echinostoma ilocanum
Common name Garrison’s fluke
Habitat Small intestine
Final host Man
Reservoir host Rat
1st IH Snail
2nd IH Snail
Egg Germ ball egg

24
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

3. Heterophyes heterophyes E. BLOOD FLUKES

Common name Von Siebold’s fluke Schistosomes
Habitat Small intestine - Adult stages are in the blood vessels
Final host Man - Most romantic parasites: “perpetual copulation”
1st IH Snail - Adult male with gynephoral canal where the female is held
2nd IH Fish - MOT: Skin penetration
Egg Resembles Clonorchis sinensis egg - Infective stage: Cercaria
- old fashioned light bulb
Adult Presence of genital sucker (Gonotyle) Disease Habitat Egg Reservoir Intermediate
caused Host
C. LUNG FLUKES S. japonicum Katayama’s Superior Small lateral Dogs, cats, Oncomelania
fever mesenteric spine carabao
1. Paragonimus westermani Swimmer’s vein of small
Common name Oriental Lung fluke itch intestine
Habitat Lungs S. mansoni Intestinal Inferior Prominent Nonhuman Biomphalaria
Final host Man bilharziasis mesenteric lateral spine primates
1st IH Snail vein of colon
2nd IH Crab, crayfish and rectum
Egg Operculated with shoulders; opposite the S. Urinary Urinary Prominent None Bulinus spp
operculum is an abopercular end haemotobium bilharziasis bladder terminal spine
Resembles D. latum egg Egyptian
Adult Resembles coffee bean hematuria
Symptoms & Endemic hemoptysis
Pathology - TB-like symptoms S. intercalatum
Laboratory Sputum - resembles S. haemotobium egg but acid-fast positive
diagnosis - Charcot-Leyden crystals - Adult located in the large intestine
S. mekongi
D. PANCREATIC FLUKE - Resembles S. mansoni egg but smaller
1. Eurytema pancreaticum - Adults located in the small intestine
Common name Pancreatic fluke
Habitat Pancreatic ducts
Final host Sheep, cattle
Laboratory Diagnosis:
Accidental Man
host a. DFS, Kato katz, FEACT
1st IH Snail b. Urinalysis
2nd IH Ant, Grasshopper, Mantis, Crickets - 24-hour unpreserved urine
Symptoms & Cholecystitis/Pancreatitits - Optimal recovery: 12PM-2PM
Pathology

25
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

c. Circumoval Precipitin Test (COPT) d. Uterine pore - Found at the medial surface
- Confirmatory test for schistosomiasis - Where eggs will exit
- Detection of antibodies that reacts with eggs - D. latum
e. Genital pore - Site where male and female organs meet
- (+) bleb formation - Where the other eggs will exit
- Specimen: serum
- Reagent: lyophilized S. japonicum eggs from a rabbit 12. Eggs: all are unoperculated and mature/embryonated except for Diphyllobothrium latum
(operculated and immature, no oncosphere and hexacanth embryo)
A. Oncosphere: protects the embryo
CESTODES B. Hexacanth embryo: embryo with 6 hooklets
General characteristics: 13. Adult: found in the intestines of the definitive hosts
1. White, yellowish, creamy white 14. Larva: encysted in the tissues of intermediate hosts
2. Adult is flat/ribbon-like
3. Segmented 15. 2 main orders:
4. Monoecious/hermaphroditic A. Order Cyclophyllidean
5. No gastrointestinal tract/alimentary canal - True tapeworm
6. No circulatory system - Species requiring vertebrate IH: Taenia solium, Taenia saginata
7. Integument: entry of nutrients and exit of wastes - Species requiring invertebrate IH: Diplydium caninum
8. Well-developed reproductive organs: testes, ovary, uterus - Species may or may not require IH: Hymenolepis nana
9. MOT: Oral route
10. Habitat: small intestine B. Order Pseudophyllidean
11. Cestode body: - False tapeworm
- Eggs are operculated and unembryonated, no oncosphere and hexacanth embryo
a. Scolex - Attaches to the lining of small intestine - D. latum and Spirometra
- Globular/pyriform or Almond/spoon-like
a. Rostellum/Crown 16. Lifespan
- Protrusible structure Up to 25 years D. latum, T. saginata, T. solium
- Armed or unarmed <1 year D. caninum, H. diminuta
b. Suckers/Grooves/Acetabula Autoinfection H. nana
- Cestodes have 4 cuplike structures except for D. latum (2
slit-like suckers) 17. Treatment: Praziquantel
c. Neck - Region of growth - Cysticercosis and hydatid disease: Albendazole, Mebendazole
- Point of proliferation for the next set of segments
d. Proglottids - Chain of segments/strobili
- Stained with a. Immature – nearest to the neck
India ink b. Mature – middle portion where reproductive parts of found
- Alchohol c. Gravid – farthest from the head; filled with eggs; can be
irritates detached (apolysis)
tapeworm

26
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Cyclophyllidean Pseudophyllidean ORDER CYCLOPHYLLIDEAN

Scolex - Globular/Pyriform - Spoon-shaped (spatula) or Almond 1. Taenia spp.
- Quadrate with 4 cuplike suckers - Slit-like sucking grooves (Bothria) Taenia saginata Taenia solium
- Armed/unarmed rostellum - No hooklets Common name Beef tapeworm Pork tapeworm
Strobila/Body - Apolytic - Anapolytic Intermediate Host Cattle, cows, camels Pig, human
- Recovered in stool: eggs & segments - Recovered in stool: eggs only Definitive Host Human Human
Ova - Spherical, unoperculated, - Oval, operculated, unembryonated Scolex None, no rostellar hooks Rostellum armed with 2 rows of
embryonated large and small hooklets
- Hexacanth embryo Length 25 meter (taller than wide) 7 meters (wider than tall)
Larval stages - Cysticercoid: D. caninum, Coracidium → Procercoid → Number of 1000-2000 Less than 1000
Hymenolepis, Rallietina Plerocercoid (Infective stage) proglottids
- Cysticercus: Taenia Gravid proglottid 15 – 20 lateral branches 7 – 13 lateral branches (dendritic
- Hydatid cyst: Echinococcus (dichotomous/tree-like) or fingerlike)
Intermediate - Only 1 IH (arthropods or mammal) - 2 IH: Eggs (reported as Spherical, striated, inside is an Spherical, striated, inside is an
host - H. nana: may or may not require IH  1st IH: Crustaceans (Copepods) Taenia egg) embryo with 6 hooklets embryo with 6 hooklets
 2nd IH: Freshwater fish Larva Cysticercus bovis Cysticercus cellulosae
Gravid - Only uterus can be seen - All reproductive structures can be Infective stage Cysticercus bovis Cysticercus cellulosae, egg (leads
Proglottid seen to cysticercosis)
- Uterus: Rosette-like Pathology Taeniasis saginata Taeniasis solium,
Uterine pore Absent Present (ventral surface, center) Neurocysticercosis
Lab Diagnosis 1. Stool Examination – perianal swab
ORDER PSEUDOPHYLLIDEAN 2. Double Slide Compression Technique
1. Diphyllobothrium latum - Use of India ink to visualize uterine branches
- Lactophenol: used to improve clearing of the specimen for
- Largest tapeworm of man
better visualization
Common name Broad Tapeworm, Fish Tapeworm
Habitat Small intestine
Final host Man Additional notes:
Reservoir host Dog, cats, rats, fish-eating mammals Taenia asiatica
1st IH Copepods (Cyclops) - 3rd Taenia spp.; reported in Taiwan, Korea, Thailand, and Indonesia
2nd IH Freshwater fish - Closely related to T. saginata,
Paratenic host Carnivorous fish - Cysticercus viscerotropica: cysticercus larvae of T. asiatica were found in the liver of
Infective stage Plerocercoid larva intermediate hosts (pigs, cattle, goats, wild boards, monkey)
Diagnostic stage Egg/Scolex
Morphology Scolex Spon-shaped/almond
*adult confused with Uterus Rosette formation (highly-coiled)
Spirometra, egg confused Egg Operculated, unembryonated
with Paragonimus With abopercular thickening
Pathology Vitamin B12 deficiency anemia

27
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

2. Hymenolepis spp. 4. Raillietina garrisoni

Hymenolepis nana Hymenolepis diminuta Habitat Small intestine
Common name Dwarf Tapeworm Rat Tapeworm Final host Rats
Habitat Small intestine Small intestine Accidental host Man
Final host Man Rat Intermediate host Flour beetle
Reservoir host Mice/rats Accidental Host: Man Infective stage Cysticercoid larva
Intermediate Direct do not require IH Insects MOT Ingestion of beetle containing larva
host Indirect Requires IH Morphology Scolex Armed rostellum with 2 alternating
(flea/beetle) hammer-shaped hooks
Infective stage Direct Embryonated egg Cysticercoid Gravid Rice grains
Indirect Cysticercoid larva proglottids
Egg Oncosphere with 2 polar Oncosphere with 2 polar thickenings Egg Enclosed in egg capsule with 1-4
thickenings from which 4-8 polar but without polar filaments “sunny spindle-shaped eggs (egg packets)
filaments arise side egg” Resembles Taenia egg
Scolex With rostellum armed with a ring With unarmed rostellum
of 20-30 spines 5. Echinococcus granulosus
Common name Hydatid worm
3. Dipylidium caninum Habitat Final host: small intestine
- The only parasite with 2 sets of reproductive organs Man/IH: connective tissues
Common name Dog Tapeworm, Double-pored Tapeworm Final host Dog
Habitat Small intestine Intermediate host Sheep
Final host Dog Accidental host Man
Accidental host Man Infective stage To Final host Hydatid cyst
Intermediate host Flea To Intermediate Egg
Infective stage Cysticercoid larva host
MOT Ingestion of flea containing larva MOT Ingestion of egg
Morphology Scolex Conical, armed rostellum Morphology Scolex Prominent armed rostellum with
4 cuplike suckers
Gravid Melon seeds, pumpkin seeds, rice Egg Cannot be differentiated from the
proglottids grains, cucumber eggs of Taenia
Egg Enclosed in egg capsule with 8-15 Lab Diagnosis 1. Roentgenogram
eggs (egg packets) *eggs are not 2. Immunologic tests
Resembles Taenia egg recovered in man a. Bentonite Flocculation test
b. Casoni’s Intradermal test
3. Surgery
- Exploratory Cyst Puncture
- Puncture Aspirate Inject Reaspirate
- 10% formalin

28
 PARASITOLOGY | Rocelle Anne R. Leonardo, RMT

Additional notes:
1. Echinococcus multilocularis
- Most lethal of all helminthic disease
- Causes alveolar hydatid disease, a fatal form of echinococcosis
- Cyst morphology is similar to that of E. granulosus, but the adult is smaller
- DH: foxes, coyotes, dogs
- IH: rodents

2. Taenia multiceps
- Causes coenurosis
- Coenurus is a unilocular cyst similar to cysticercus, although the worm has multiple
scolices
- Adult worm is typically found in dogs and other canids
- IH: sheep, cattle, deer, humans (accidental ingestion of dog feces containing the
eggs)

29