2. Diarrheal Disease due to Parasites
Travel history or poor sanitation put you at the highest risk for
parasitic infection
Poor immune status higher risk
Parasitic infections associated with sporadic symptoms
Dysentery not common (usually only amebiasis)
Most usual symptoms:
◦Abdominal pain, cramping, long term nausea, and malaise, mucous in stool,
and +/- fever, chronic in nature
3. Laboratory Diagnosis
Current diagnosis is predominately
based on microscopic exam
Specimen types:
◦Stool
◦Non-stool
◦Perianal specimen
◦Sigmoidoscopic specimen
◦Duodenal aspirates
◦Liver abscess
◦Sputum
◦Urine
◦Urogenital
◦Blood
◦Tissue
Alternative testing methods:
◦Serology for select pathogens
◦Fluorescent stains of stool for select
pathogens
◦Molecular assays of stool for select
pathogens
4. From the 2017 IDSA Practice
Guidelines: suggested
laboratory testing methods for
select pathogens
Parasite Test Method Specimen
5. Two-vial collection kit for stool
preservation for routine parasite
exam
10% formalin vial
Concentration of stool performed
with ethyl acetate washing to
eliminate fecal debris
Can use to perform: Wet mount,
Iodine mount, DFA staining and
NAAT
Detect: Helminth eggs, larvae,
microsporidia, and protozoan cysts
PVA with fixative
Polyvinyl alcohol (PVA)
Permanent smear
prepared from stool and
stained with Trichrome
stain
Detect: Protozoan
trophozoites and cysts
9. Entamoeba histolytica/dispar
E. histolytica (pathogen) and E. dispar (nonpathogen) species both occur in
the large intestine. Morphologically indistinguishable.
◦Use either antigen testing or molecular methods to distinguish the two
species
E. histolytica in the intestine
◦Cysts = infectious form found in the environment
◦Contaminated water and poor sanitation
◦Trophozoites = invasive form in the intestine
◦Colon biopsy shows “flask-shaped” ulcers with trophozoites
Extra-intestinal disease
◦Liver abscess – amebiasis with trophozoites in liver biopsy
◦Serology will be positive in most cases compared to <=50% of cases of
intestinal only infection
12. Amebic abscess
Flask-shaped ulcer of intestinal amebiasis
Amebic liver abscess/ flask shaped ulcer
E. histolytica serology – high % positive
in extra-intestinal cases
trophozoite
13. Entamoeba coli
commensal in the intestine
Cyst @ 20 – 25µm
Up to 8 nuclei
Shed from host
Lives in environment
Trophozoite is the
invasive form that is
found in the
intestine
Single nucleus with a large
karyosome located
eccentrically with irregular
chromatin ring (differs from E.
histolyica).
The cytoplasm appears dirty
and vacuolated
14. Entamoeba coli – important to
differentiate from the pathogen E. histolytica
Trophozoites & Cysts
Cysts usually
15-25µm, with 5
or more nuclei
visible.
15. Endolimax nana
Trophozoites 8 – 10 µm, one
nucleus
Cysts – 6-8 µm in size, four
nuclei
Usually a commensal, seen in
stool specimens from HIV/AIDS
patients, some literature
suggesting it can cause
intermittent or chronic diarrhea
16. Iodamoeba butschlii
Cysts, 10 – 12 µm
Large starch inclusion
(glycogen mass) that is unique
to Iodamoeba
Usually not considered a pathogen
Iodine preparation – glycogen
inclusions stain with iodine
19. Giardia lamblia (G.
intestinalis,
and G. duodenalis)
Found in contaminated water & undercooked
foods
Mild diarrhea to severe malabsorption
Foul smelling , watery diarrhea
Day-care center outbreaks reported, traveler’s
diarrhea
Cysts/trophozoites may be seen in stool, but
can be difficult to detect; Fluorescent stains
and NAAT for more sensitive detection
Duodenal aspirations can be used if stool
specimens are negative
TROPHOZOITE
“falling leaf” motility
CYSTS
20. Giardia lamblia trophozoite
Waxing and waning
symptoms
Can be irregularly
shed in stool material
making antigen
and molecular methods
more sensitive for
detection
Russia & Mexico
-Hot beds of infection
Confined to intestine –
does not disseminate
Flagyl (Metronidazole)
is drug of choice
22. Giardia lamblia
only invades intestinal tissue – shown are duodenal biopsies,
trophozoites near the surface epithelium
23. Chilomastix mesnili cyst
Nonpathogen
Morphology mimics Giardia
lamblia cyst – except for the clear
space at end of cyst
Internal structure looks like
“shepherd’s crook” or safety pin
C. mesnili trophozoite – seldom
seen in clinical specimens
24. Dientamoeba fragilis
Diarrhea, anal pruritus
Only a trophozoite stage 5 – 15 µm (No
cyst)
Usually two nuclei visible in the
trophozlite
Can occur in Co-infection with Enterobius
(pinworm)
25. Trichomonas vaginalis
Urogenital protozoan
STD in both men and women
Infection leads to Scant, watery
vaginal discharge in women
Infection in urethral orifice in men
Four flagella, short undulating
membrane
27. Ciliates
Balantidium coli
◦Contact with swine
◦Poor hygiene
◦Only ciliate that’s pathogenic to humans
◦Similar disease as amebiasis
◦Largest (50-200 um) trophozoite in parasitology
◦Surface covered with cilia; macronucleus
◦Cyst 40-60 um
◦Readily identified in fresh, wet mounts
◦Can cause flask shaped ulcer in intestine like that of
Entamoeba histolytica
28. Isospora(Cystoisospora)
belli
Contaminated food/water, oral-anal route of infection
Found most commonly in HIV/AID patients
Infects intestinal epithelium
Malabsorption syndrome mimicking giardiasis
Stains positive with modified acid fast staining
(+) Modified acid fast stain
29. Cryptosporidium spp
Contaminated water
◦Resistant to usual water-purification procedures
(chlorination, ozone)
Watery diarrhea -
◦More severe in HIV/AIDS – chronic/debilitating
infection
◦Daycare center outbreaks (fecal-oral transmission)
Not detected in routine O & P exams (left)
Modified acid-fast stains (PAF) aid in detection,
Oocysts measure 4-6 µm
Stool antigen, Direct Fluorescence antibody staining
and Molecular assays aid detection.
30. Cryptosporidiosis: Diagnosis
Not detected in routine O & P exams (left)
Requires modified acid-fast stains for detection, oocysts
measure 4-6 µm , Antigen, DFA and Molecular assays aid
detection.
PAF stain
positive
Direct Fluorescence Antibody stain –
Cryptosporidium spp
32. Cyclospora
cayetanensis
Contaminated fruits and vegetables – particularly ones with plant hairs
Watery diarrhea; fatigue, anorexia, weight loss, flu like symptoms. More severe in
immune suppressed, can last for months
Infects upper small bowel
Treatment Oral Trimethoprim/sulfamethoxazole
Found in vacuoles in cytoplasm of jejunal epithelium, villous atrophy, crypt
hyperplasia
Modified acid
fast positive
Cysts 8-10
microns
UV autofluorescence
Also positive on Calcofluor
white stain
33. Microsporidia
Obligate intracellular fungal parasite (yeast like)
Enterocytozoon and Encephalitozoon species most
common genera
Infection by ingestion of spores
Chronic diarrhea in HIV/AIDS patients
Can also disseminate and be found in cases of
Myositis, hepatitis, peritonitis, keratitis,
gastrointestinal and biliary tract infection
34. Microsporidia
-Diagram of detailed internal spore
structures
Spores are 1 – 4um
Stain positive on Calcofluor white and
modified Trichrome stains – modified
by longer staining times that eventually
allow for stain to penetrate the spore
35. Blastocystis hominis
(algae)
Small #s: are considered commensal
Large #s: may be pathogenic
Found in Contaminated food and H20
Primary a cause of Traveler’s diarrhea
Iodine wet mount
Nuclear blobs
Around the periphery Trichrome
stain
36. None;None;
self resolving.self resolving.
Maltese cross in rbcMaltese cross in rbcHemolytic anemia,Hemolytic anemia,
Jaundice, fever,Jaundice, fever,
hepatomegalyhepatomegaly
Ixodes tickIxodes tickBabesia microtiBabesia microti
Pentosam;Pentosam;
PentamidinePentamidine
isethionate.isethionate.
IntracellularIntracellular
(macrophages)(macrophages)
leishmanial bodiesleishmanial bodies
with kinetoplastwith kinetoplast
Visceral leishmaniasisVisceral leishmaniasis
(Kala-azar),(Kala-azar),
granulomatous skingranulomatous skin
lesionslesions
Iraq/Iran/AfghanistanIraq/Iran/Afghanistan
Phlebotomine sandflyPhlebotomine sandflyLeishmania donovaniLeishmania donovani
CNS:CNS:
melarsoperolmelarsoperol
Nifurtimox andNifurtimox and
Benzonidazole.Benzonidazole.
Hemoflagellate inHemoflagellate in
blood or tissue.blood or tissue.
C- or comma-shapedC- or comma-shaped
AmericanAmerican
trypanosomiasis;trypanosomiasis;
Chagas disease:Chagas disease:
megacolon, cardiacmegacolon, cardiac
failure.failure.
Reduvid (kissing) bugReduvid (kissing) bugT. cruziT. cruzi
Blood stage:Blood stage:
Suramin orSuramin or
petamidinepetamidine
isethionateisethionate
Hemoflagellate inHemoflagellate in
blood or lymph nodeblood or lymph node
AfricanAfrican
trypanosomiasis;trypanosomiasis;
Sleeping sicknessSleeping sickness
Encephalitis; cardiacEncephalitis; cardiac
failurefailure
Tsetse flyTsetse flyTrypanosoma bruceiTrypanosoma brucei
TreatmentTreatmentDiagnosisDiagnosisDisease/SymptomsDisease/SymptomsTransmissionTransmissionOrganismOrganism
BLOOD BORNE PROTOZOA
37. Trypanosoma cruzi Chagas disease
(American trypanosomiasis)
Vector: Reduvid/Triatoma (kissing) bug
Trypomastigote is the form in the blood of an infected person; may be seen in
CSF in CNS infections
Motile circulating trypomastigotes are readily seen on slides of fresh
anticoagulated blood in acute infection but are rarely detectable by
microscopy in chronic T. cruzi infection.
A typical trypomastigote has:
◦ A large, subterminal or terminal kinetoplast,
◦ A centrally located nucleus,
◦ An undulating membrane, and
◦ A flagellum running along the undulating membrane, leaving the body at the
anterior end.
◦ 12 to 30 µm in length.
◦ Can appear C shaped
Amastigote stage parasite may be seen in histopathology specimens from
affected organs.
Trypomastigote
Amastigote
38. Trypanosoma brucei Sleeping sickness
(African trypanosomiasis)
Vector: Tse Tse fly
The two T. brucei species that cause African trypanosomiasis are
indistinguishable morphologically
◦ T. brucei gambiense
◦ T. brucei rhodesiense
A typical trypomastigote has:
◦ A small kinetoplast located at the posterior end
◦ A centrally located nucleus
◦ An undulating membrane, and
◦ A flagellum running along the undulating membrane, leaving the
body at the anterior end
◦ 14 to 33 µm in length
Trypomastigote is the only stage found in specimens
kinetoplast
nucleus
40. Leishmania – Clinical
Disease
Cutaneous
◦Single or few chronic, ulcerating lesions; many species
◦Latin America, southern Europe, Middle east,
southern Asia, Africa
◦Mucocutaneous in Latin America
Visceral
◦primarily L. donovani complex (Asia), L.
infantum/chagasi (Africa and Latin America), others
◦Hepatosplenomegaly, anemia, cytopenias, systemic
symptoms
◦India, Bangladesh, Nepal, Sudan, and Brazil
◦Important opportunistic infection in HIV infection
41. Leishmania
Diagnosis
◦Biopsy of infected tissue (skin, bone marrow)
◦Multiple, tiny 2-5 um amastigotes within histiocytes
◦Distinct kinetoplast (bar-like structure adjacent to nucleus)
◦PCR methods
◦Urinary antigens (visceral)
DDx of multiple tiny intracellular organisms
◦Leishmania – kinetoplast
◦Histoplasma – budding
◦Toxoplasma – somewhat curved, mostly
extracellular
42. Babesia
Protozoan: B. microti, B. divergens
Zoonosis (deer, cattle, rodents; humans accidental host)
Transmission vector: Ixodes tick bite
Infects red blood cells
Found world-wide
B. microti along the Northeast US
◦ Nantucket Island, Martha’s vineyard, Shelter Island
Malaria-like syndrome
◦ Fever but without periodicity, night sweats, weight loss,
hemolytic anemia, hemoglobinuria, renal failure
Dx:
◦ Blood smear examination
◦ Ring form only (mimics P. falciparum)
◦ Tetrads (unlike P. falciparum)
Maltese cross
(tetrads)
Ixodes tick
46. Life Cycle of Plasmodium Species
Ring form Trophozoite Schizont Merozoite
Ruptured Schizont
Gametocyte
47. Malarial Preparations
Thick smear
Drop of blood on slide (non-
anticoagulate blood is best)
Water rinse to eliminate rbc’s
Stain with Giemsa stain (not
Wright-Giemsa) proper pH
Need the proper pH to stain
the Schuffner’s granules
Concentrated to spot malaria
parasites
Thin smear
Feather edge smear
For optimal morphology, stain
with Giemsa (not Wright-
Giemsa) stain with proper pH
Speciation of malaria
Parasitemia (%)
49. P. vivax
Amoeboid
ring form
P.vivax – benign tertian malaria (fever every 48 hours), invades reticulocytes.
Duffy negative Red blood cell protects from Plasmodium vivax invasion-
African natives lack Duffy rbc antigen and this prevents them from serious P. vivax.
Untreated infections last several years remaining dormant in the liver.
Recurrent and chronic infection can lead to brain, kidney and liver damage
54. P. falciparum
Multiple ring forms per cell can be seen
Accolade forms(ring forms on the edge of
cell)
No schizonts circulate in blood
Banana shaped gametocyte
High % parasitemia
Calculate % parasitemia
No. infected RBCs + total no. RBCs counted X 100 = % infected RBCs
55. Toxoplasma
gondii
T. gondii encystment in tissue T. gondii tachyzoites
Toxoplasmosis is considered to be a leading cause of death attributed to foodborne illness in the United
States. Caused by eating undercooked, contaminated meat (especially pork, lamb, and venison)
Accidental ingestion of oocysts after cleaning a cat's litter box when the cat has shed Toxoplasma in its
feces is also a cause
More than 30 million men, women, and children in the U.S. carry the Toxoplasma parasite, few have
symptoms the immune system keeps the parasite from causing illness.
Women newly infected with Toxoplasma during pregnancy (TORCH) and anyone with a compromised
immune system are most at risk for severe consequences.
57. Primary Amoebic meningoencephalitis (PAM) is
caused by Naegleria fowleri
Granulomatous amoebic encephalitis (GAE) is caused
by Acanthamoeba and Balamuthia
Clinical scenario: swimming or diving into fresh-water pools
and water goes up nose thru cribriform plate
N. fowleri
trophozoite
58. Contact-lens keratitis
Cause: Acanthamoeba
◦To culture Acanthamoeba, Corneal scrapings placed in a lawn of
E.coli
◦Visible tracks of ameba detected on agar plate
◦Wright stain of corneal scrapings is simpler/better way
Wright’s
stain
Amoeba in
wet mount
60. Nematodes (Roundworms)
Enterobius vermicularis
Ascaris lumbricoides
Trichuris trichuria
Necator americanus & Ancylostoma duodenale
Strongyloides stercoralis
Anisakis simplex
Microfilaria – Wucheria, Brugia, Loa loa, Mansonella, and
Onchocerca
62. Enterobius vermicularis
(pinworm)
Humans / only host
Most common helminth in US
Worms: Females 8-13mm, males 2-5 mm
◦Appear like strings in the stool
Dwell in the cecum
Migrate to perineum at night
◦Deposit eggs
◦Diagnosis- Scotch tape test or anal swab in AM
Oval with a flattened side: 50-60um by 20-30um
63. Ascaris lumbricoides
(roundworm)
1-1.2 billion people infected, Africa, the Americas, China and East Asia,
common in children
Developing countries with poor sanitation/ feces contaminated soil,
humans ingest eggs
Largest helminth to affect humans
◦Worms: Females 20-35cm long and straight, males 15-30cm with a curved
tale
◦Can cause intestinal obstruction
◦Loeffler’s syndrome – pulmonary infiltration and eosinophilia from
worm migration
64. Ascaris lumbricoides
Charcot Leyden crystals are hexagonal bipyramidal
structures localized in the primary granules of the
cytoplasm of eosinophils. Their presence is indirect
evidence of parasitic infestation particularly with
Ascariasis, Toxocara, Capilliriasis, or Fasciola.
Bolus of Ascaris
removed from
Bowel obstruction
66. Trichuris trichuria
(whipworm)
Soil transmitted/fecal contamination
Disease similar to amebiasis
PVA preserved samples inferior to formalin
Adult worm attaches to large intestine, rarely
recovered – diagnosis by detecting egg in stool
specimens
Thinnest part of worm is head
Egg is barrel shaped, golden brown, knobs on both
ends
67. Necator americanus and Ancylostoma
duodenale (Hookworms)
Soil transmitted – filiform larvae penetrate the skin
2nd
most common helminth infection
Necator or Ancylostoma – Hookworm egg
Both genera egg looks alike – egg
60 X 40 um
68. Strongyloides stercoralis
Soil transmitted
Larval form only –
◦Penetrate intact skin
Usually intestinal larval infestation
In immune suppressed - massive intestinal
infection with +/- migration to the respiratory tract
(eosinophilic pneumoniae) so called autoinfection
Strongyloides larvae
69. Anisakis simplex
Anisakiasis is a parasitic disease caused by anisakid nematode (worms) that can invade the
stomach wall or intestine of humans.
The transmission of this disease occurs when infective larvae are ingested from fish or squid
that humans eat raw or undercooked.
In some cases, this infection is treated by removal of the larvae via endoscopy or surgery. Can
in rare cases cause granuloma formation and obstruction.
71. Microfilariae
Sheathed
◦Wucheria bancrofti and Brugia malayi
◦Elephantiasis (lymphangitis/lymphedema)
◦Loa loa
◦Calabar swellings & migrating worms in the conjunctiva
Not sheathed
◦Onchocerca volvulus
◦Mansonella species
◦Allergic skin reactions, edema, Calabar swellings
73. Identification of microfilariae is based on the presence of a sheath covering the larvae, as well
as the distribution of nuclei in the tail region
A, W. bancrofti. B, B. malayi. C, L. loa. D, O. volvulus. E, Mansonella perstans. F, Mansonella
streptocerca. G, Mansonella ozzardi.
74. Filaria
Identification
a. W. bancrofti
◦ Sheathed, nuclei stop short of end
of tail
a. B. malayi
◦ Sheathed, two small nuclei in tail
a. O. volvulus
◦ Unsheathed, from skin, not blood
a. Loa loa
◦ Sheathed, nuclei to continue to
end of tail
84. Fasciola hepatica
Distinct nose
Fascioliasis- Fasciola hepatica, "the common liver fluke" or "the sheep liver fluke.
Associated with sheep & cattle
Infected by eating raw watercress or other water plants contaminated with immature larvae.
The immature larval flukes migrate through the intestinal wall, the abdominal cavity, and the liver
tissue, into the bile ducts. The pathology is most pronounced in the bile ducts and liver.
Fasciola infection is both treatable and preventable
Eggs – ellipsoidal, operculated and large 140 X 80µm
85. Fasciolopsis buski, causes fasciolopsiasis, is the largest intestinal fluke of humans.
Prevented by cooking aquatic plants well before eating them.
Found in south and southeastern Asia.
Fasciolopsiasis is treatable.
Many people do not have symptoms from Fasciolopsis infection. However, abdominal pain
and diarrhea can occur 1 or 2 months after infection.
With heavy infections Fasciolopsis flukes can cause intestinal obstruction, abdominal pain,
nausea, vomiting, and fever.
87. Egg is operculate, not
embryonated, thick shell,
asymmetrical and large
Paragonimus westermani
Paragonimus is a parasitic lung fluke.
Infections occur after a person eats raw or
undercooked infected crab or crayfish.
Paragonimus infection also can be very
serious if the fluke travels to the central
nervous system, where it can cause
symptoms of meningitis.
89. Schistosomiasis, also known as bilharzia, more than 200 million people are infected
worldwide. Second only to malaria as the most devastating parasitic disease.
Live in certain types of freshwater snails. The infectious form of the parasite, known as
cercariae, emerge from the snail, and contaminate water. You become infected when your
skin comes in contact with contaminated freshwater. Most human infections are caused by
(1)Schistosoma mansoni, (2) S. haematobium, or (3) S. japonicum.
Examine Stool for S. mansoni or S. japonicum eggs and Urine for S. haematobium
eggs).
1 2 3
91. Cestodes (Tapeworms)
Examples
Diphyllobothrium latum
Taenia saginata
Taenia solium
Hymenolepis nana
Hymenolepis diminuta
Echinococcus granulosis
Head with armed or unarmed scolex
Proglottids – tapeworm elongates by
producing Proglottids
92. Diphyllobothrium
latum
Infected by ingesting poorly-cooked fresh-water fish
(salmon particularly problematic)
Scandinavian, Russia, Canada, N. USA, Alaska
Known as the Broad fish tapeworm
Scolex has a Longitudinal sucker
Eggs have non-shouldered operculum and knob
◦They are not embryonated
Infection causes VitaminB12 deficiency
94. Taenia saginata
Beef tapeworm
4 suckers on scolex
>12 uterine branches in
proglottids
Ingestion of cysticerci in
beef
Intestinal tapeworm
Ingestion of eggs ->
Non-human pathogen
No disease
Taenia Solium
Pig tapeworm
Ring of thorns/crown on
scolex
<12 uterine branches in
proglottids
Ingestion of cysticerci in
pork
Intestinal tapeworm
Ingestion of eggs ->
Cysticercosis
100. Hymenolepis nana (Dwarf Tapeworm)
Definitive host: Human and rodent
Worm is 2-4 cm – shortest of the tapeworms
Egg has inner & outer shell separated space
Water /food contaminated by rodent droppings
102. Echinococcus – hydatid cyst disease
found in Africa, Europe, Asia, the Middle East, and Central
and South America.
Highest prevalence is found in populations that raise sheep.
Infection from ingestion of eggs found in animal feces.
103. Echinococcus – hydatid cyst
Short tapeworm
Sand like material
contained in the
Hydatid Cyst, due to
inverted folded
tapeworms
Hydatid cyst in liver
104. Relative size of Helminth
eggs
http://www2.bc.cc.ca.us/bio16/pal/Parasitology.htm
106. Maggots
House fly larvae
Bot fly larvae
Bot fly bites human,
larvae develops in
subcutaneous area, matures
and then
extrudes from the skin
Native to central and south
America
Bot fly
Particularly difficult to differentiate from falciprium
Blackwater fever (falciprium and think black pee)
Microscopic examination
The malaria parasite life cycle involves two hosts. During a blood meal, a malaria-infected female Anopheles mosquito inoculates sporozoites into the human host. Sporozoites infect liver cells and mature into schizonts, which rupture and release merozoites. (Of note, in P. vivax and P. ovale a dormant stage [hypnozoites] can persist in the liver and cause relapses by invading the bloodstream weeks, or even years later.) After this initial replication in the liver (exo-erythrocytic schizogony ), the parasites undergo asexual multiplication in the erythrocytes (erythrocytic schizogony ). Merozoites infect red blood cells . The ring stage trophozoites mature into schizonts, which rupture releasing merozoites. Some parasites differentiate into sexual erythrocytic stages (gametocytes). Blood stage parasites are responsible for the clinical manifestations of the disease. The gametocytes, male (microgametocytes) and female (macrogametocytes), are ingested by an Anopheles mosquito during a blood meal . The parasites’ multiplication in the mosquito is known as the sporogonic cycle . While in the mosquito&apos;s stomach, the microgametes penetrate the macrogametes generating zygotes . The zygotes in turn become motile and elongated (ookinetes) which invade the midgut wall of the mosquito where they develop into oocysts . The oocysts grow, rupture, and release sporozoites , which make their way to the mosquito&apos;s salivary glands. Inoculation of the sporozoites into a new human host perpetuates the malaria life cycle .
Most prevalent
Widest geographical distribution
P
Enlarged RBC; fimbriated/ragged rbc
Fever cycle every 72 hours (quartan), can remain dormant in the blood for years.
Untreated infections may last as long as 20 years