2. Clinical presentation
Travel history or poor sanitation put you at the
highest risk for parasitic infection
Infections associated with sporadic symptoms
Poor immune status higher risk
Dysentery not common (amebiasis)
Most usual symptoms:
◦Abdominal pain, cramping, long term nausea, and
malaise, mucous in stool, and +/- fever
3. Laboratory Diagnosis
Currently based on microscopic exam,
and molecular panels for the most
common (select) parasites
◦Stool (PCR applicable)
◦Non-stool
◦ Perianal specimen
◦ Sigmoidoscopic specimen
◦ Duodenal aspirates
◦ Liver abscess
◦ Sputum
◦ Urine
◦ Urogenital
◦Blood
◦Tissue
Alternative methods:
◦Serology
◦Fluorescent stains
4. Two-vial collection kit for
Stool
10% formalin
Concentration with ethyl
acetate to eliminate fecal
debris
Wet mount, DFA staining and
NAAT
Helminth eggs, larvae,
microsporidia, and protozoan
cysts
PVA with fixative
Polyvinyl alcohol
Permanent stained smear
◦Trichrome stain
Protozoan trophozoites and
cysts
Mercury based fixatives
phased out for safety – Zinc
fixatives are now used
9. Entamoeba
histolytica/dispar
E. histolytica is a pathogen and E. dispar is a nonpathogenic
species that can also occur in the large intestine.
Morphologically indistinguishable
◦Antigen testing or molecular methods to distinguish the two species
E histolytica
◦Cysts = infectious form
◦Trophozoites = invasive form
◦Cysts found in contaminated water and poor sanitation
◦Colon biopsy shows “flask-shaped” ulcer with Trophs
◦Non-intestinal disease = extra-intestinal amebiasis (liver
abscess) with Trophs
◦Serology can be useful
12. Amebic abscess
Flask-shaped ulcer of intestinal amebiasis
Amebic liver abscess
Entamoeba histolytica
Serology – high %
positive in extra-
intestinal cases
13. Entamoeba coli cyst and trophozoite
Considered a commensal in the
intestine
Cyst @ 20 – 25µm
Up to 8 nuclei
Shed from host
Lives in environment
Trophozoite is the
invasive form that
invades the
intestine
Single nucleus with a large
karyosome located eccentrically
irregular chromatin ring.
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 trophozoite
Cysts – 8-10 µm in size, one nucleus
Mostly thought to be a non-pathogen, 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 in
size with starch inclusion (glycogen mass)
Iodine preparation – glycogen
inclusion stains with iodine
19. Giardia lamblia
Contaminated water,
undercooked foods
Mild diarrhea to severe
malabsorption
Foul, watery diarrhea
Day-care center outbreaks
reported, traveler’s diarrhea
Cysts/trophozoites may be seen in
stool, but can be hard to find;
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
necessary for detection
Russia & Mexico
-Hot beds of infection
Confined to intestine
Flagyl (Metronidazole)
is drug of choice
24. Chilomastix mesnili cyst
Nonpathogen
Mimics Giardia lamblia cyst –
except for clear space at end of
cyst
Internal structure looks like
“shepherd’s crook” or safety pin
C. mesnili trophozoite
25. 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)
31. Cryptosporidium spp
C. parvum and C. hominis
Contaminated water
Resistant to usual water-purification procedures
(chlorination, ozone)
Daycare center outbreaks (fecal-oral), swimming
pools
Watery diarrhea; more severe in AIDS
32. Cryptosporidiosis: Diagnosis
Partial Acid Fast Stain Positive
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
33. Direct Fluorescence Antibody stain –
Cryptosporidium spp
Molecular assays (PCR) and
Enzyme immunoassay for antigen
available.
Giardia
Cryptosporidium
Combo stain for Cryptosporidium
and Giardia lamblia
35. 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
37. Microsporidia
Obligate intracellular fungal parasite
Enterocytozoon and Encephalitozoon species most
common genera
Primitive eukaryotic organism (fungi)
Infection by ingestion of spores
Chronic diarrhea in AIDS patients
Myositis, hepatitis, peritonitis, keratitis,
gastrointestinal and biliary tract
38. Microsporidia
-Diagram of detailed internal spore
structures
Positive on modified Trichrome
and Calcofluor white stains
-Longer staining times will eventually
allow for it to work its way into the spore
39. Blastocystis hominis (algae)
Small #s: can be commensal
Large #s: may be pathogenic
Contaminated food and H20 Traveler’s diarrhea
Iodine wet mount
Nuclear blobs
Around the periphery Trichrome
stain
40. 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
42. Trypanosoma cruzi Chagas
(American trypanosomiasis)
Vector: Reduvid/Triatoma (kissing) bug
Trypomastigotes are the only stage found 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.
Amastigote stage parasite may be seen in histopathology
specimens from affected organs.
C-shape
44. Trypanosoma brucei
Sleeping sickness (African
trypanosomiasis)
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
Trypomastigotes are the only stage found in patients. kinetoplast
nucleus
48. 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 OI in HIV infection
49. 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
51. 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, “B-symptoms”, hemolytic anemia,
hemoglobinuria, renal failure
Dx:
◦ Blood smear examination
◦ Ring form only (mimics P. falciparum)
◦ Tetrads (unlike P. falciparum)
Maltese cross
(tetrads)
Ixodes tick
55. Malaria
Physical exam findings
◦Fever
◦Splenomegaly
◦P. falciparum - most pathogenic species
◦Jaundice
◦Hepatomegaly
◦Increase in respiratory rate
◦CNS involvement
◦Blackwater fever – hemolysis, renal failure
Diagnosis: peripheral blood smear
Antigen screen (EIA) and NAAT
56. Malaria
Primary distinction is P. falciparum and non-falciparum
disease
◦P. falciparum = rapidly progressive and LETHAL (malignant
tertian fever), often chloroquine-resistant
◦Non-falciparum = rarely cause severe manifestations, often
chloroquine sensitive
Relapsing malaria
◦Dormant hepatic phase can recur from liver
◦Occurs only with P. vivax and P. ovale
61. Malarial Preparations
Thick smear
Drop of blood on slide
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 (%)
67. P. vivax
Amoeboid ring form
P.vivax – benign tertian malaria (fever every 48 hours),
Duffy negative Red blood cell protects from Plasmodium invasion-
African natives lack Duffy rbc antigen and this prevents them from P. vivax.
Untreated infections last several years remaining dormant in the liver.
Patients can survive years without treatment, but recurrent and chronic infection can
lead to brain, kidney and liver damage
68. • P. falciparum
-multiple ring forms
per cell
-accolade forms(ring
forms on the edge of
cell.
-Banana gametocyte
77. Amoebic
meningoencephalitis
Most common cause: Naegleria fowleri
Granulomatous amoebic encephalitis or brain
abscess: Acanthamoeba and Balamuthia
Clinical scenario: swimming or diving into fresh-
water pools
79. Contact-lens keratitis
Caused by Acanthamoeba
Can be cultured on a “lawn of E. coli”
◦Take corneal scrapings
◦Visible trail of ameba moving across plate ingesting
E. coli
Wright’s stain
83. Enterobius vermicularis
(pinworm)
Humans considered only host
Females 8-13mm, males 2-5 mm
Dwell in the cecum
¼-1/2 inch in thickness, white, lloks like string in
stool
Lay up to 15,000 eggs
◦Oval with a flattened side: 50-60um by 20-30um
Diagnosis- Scotch tape test or anal swab
Most common helminth in US
86. Ascaris lumbricoides
(roundworm)
1-1.2 billion people infected
◦More common in children
20,000 death
Largest helminth to affect humans
Females 20-35cm long, males 15-30cm with a
curved tale
◦Can cause intestinal obstruction
89. Trichuris Trichiura
(whipworm)
Soil transmitted
Disease can can be similar to amebiasis
PVA preserved samples inferior to formalin
Adults attach to large intestine and are rarely
recovered – diagnosis by detecting egg in stool
specimens
Thinnest part- head
Males are smaller than females
92. Strongyloides stercoralis
Soil transmitted
Larval form only
Presence of internal structures can separate from
artifact
In immune suppressed - massive intestinal infection and
migration to the respiratory tract (eosinophilic
pneumoniae) possible
Strongyloides
larvae
95. 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
97. 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.
98. 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
108. Fasciola hepatica
Distinct nose
Fascioliasis is a parasitic infection typically caused by Fasciola hepatica,
also known as "the common liver fluke" or "the sheep liver fluke.
Fascioliasis is found in all 5 continents, especially where sheep or cattle
are reared. Infected by eating raw watercress or other water plants
contaminated with immature parasite larvae. The immature larval
flukes migrate through the intestinal wall, the abdominal cavity, and
the liver tissue, into the bile ducts, where they develop into mature
adult flukes, which produce eggs. The pathology typically is most
pronounced in the bile ducts and liver. Fasciola infection is both
treatable and preventable
109. Intestinal fluke 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.
111. Egg is operculate, not
embryonated, thick shell,
asymmetrical and large
Paragonimus westermani
Paragonimus is a parasitic lung fluke (flat
worm). Infections occur after a person
eats raw or undercooked infected crab or
crayfish. The illness is known as
paragonimiasis. Paragonimus infection
also can be very serious if the fluke
travels to the central nervous system,
where it can cause symptoms of
meningitis.
113. Schistosomiasis, also known as bilharzia, more than 200 million
people are infected worldwide. In terms of impact this disease is
second only to malaria as the most devastating parasitic disease.
The parasites that cause schistosomiasis live in certain types of
freshwater snails. The infectious form of the parasite, known as
cercariae, emerge from the snail, hence contaminating water. You
can become infected when your skin comes in contact with
contaminated freshwater. Most human infections are caused by
Schistosoma mansoni, S. haematobium, or S. japonicum.
Stool or urine samples can be examined microscopically for parasite
eggs (stool for S. mansoni or S. japonicum eggs and urine for S.
haematobium eggs).
114. Cestodes (Tapeworms)
Examples
Diphyllobothrium latum
Taenia saginata
Taenia solium
Hymenolepis nana
Hymenolepis diminuta
Echinococcus granulosis
Flattened dorsoventrally, segmented
Head with armed or unarmed scolex
Proglottids immature, mature (sex organs)
Gravid (with eggs)
Internal structure of proglottids
Hermaphroditic-ovary, testes, vitellaria,
uterus, genital pore and ducts
Lateral excretory and nervous system
No gut-tegument absorbs nutrients
Muscles-longitidinal and horizontal
116. Diphyllobothrium latum
Poorly-cooked fresh-water fish (salmon)
Scandinavian, Russia, Canada, N. USA, Alaska
Broad fish tapeworm
Longitudinal sucker
Eggs have non-shouldered operculum and knob
◦They are not embryonated
Infection causes VitaminB12 deficiency
119. Taenia saginata
Beef tapeworm
4 suckers on scolex
>13 uterine branches in
proglottids
Ingestion of cysticerci in
beef
Intestinal infestation
Ingestion of eggs ->
Non-human pathogen
Taenia Solium
Pig tapeworm
Ring of thorns/crown on
scolex
<13 uterine branches in
proglottids
Ingestion of cysticerci in pork
Intestinal infestation
Ingestion of eggs ->
Cysticercosis
Taenia Species – two species
Outstanding characteristics
127. Hymenolepis nana
Most common cestode recovered in USA
Worm is 2-4 cm
Egg has inner & outer shell separated
space
Water /food contaminated by rodent
droppings
Hooklets inside
Larger outer shell
No radial striations
129. 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 ingestion of
egg found in animal feces.
130. Echinococcus – hydatid cyst
Short tapeworm
Sand like material
contained in the
Cyst, due to inverted folded
tapeworms
131. Relative size of Helminth
eggs
http://www2.bc.cc.ca.us/bio16/pal/Parasitology.htm
Particularly difficult to differentiate from falciprium
Hallmark of babesiosis is the tetrad (Maltese cross; cruciform body)
Blackwater fever (falciprium and think black pee)
Microscopic examination
Exception for non-falcirpium is P. vivax where chlorquine resistance is seen in Papua New Guinea and Indonesia
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 .
Presence of absence of various stages in the blood
Morphology of the gametocyte
Size of the infected RBC
Most prevalent
Widest geographical distribution
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