Pathogenic Protozoa: Intestinal Parasites Flashcards
Protozoa are unicellular ____
Eukaryotes
Do protozoa have cell walls? If not, what do they have?
NO! Cyst forms have a cyst wall
Three types of protozoan locomotion appendages
- Cilia
- Flagella
- Pseudopodia
These are temporary, foot-like projections filled w/ cytoplasm in amoeboids; mediate “crawling” over surfaces
Psuedopodia
These are hairlike projections that typically cover the surface of ciliates; beat in waves that propels a cell
Cilia
These are flexible, whip-like projections localized to specific areas on flagellates; movement propels a cell
Flagella
Motile, feeding, and proliferative form of protozoa
Trophozoite (“troph”)
Most protozoa produce a ____ form in response to adverse environmental stimuli
Cyst
What is the purpose of the cyst form?
Cyst encases the troph w/in a thick shell to protect from harsh conditions
Process of forming a cyst
Encystation (enter)
Process of leaving a cyst
Excystation (exit)
For many pathogenic protozoa, what is the infectious form?
Cysts
In fecal concentrates, ____ are damaged and unrecognizable, ____ remain intact and are observable
Trophs; cysts
Three lab methods for detecting protozoa in stool specimens
- Ag detection assays
- Microscopic examination
- Molecular methods
Which form of protozoa are found more in formed stool? Watery stool?
- Formed: cysts
- Watery: trophs
Why should more than 1 fecal specimen be examined?
B/c of intermittent shedding of parasites in stool
How should stool be collected for detecting protozoa?
Three specimens over the period of 10 days
How long should you wait to preserve fecal specimens?
DON’T WAIT! Preserve immediately!
Disadvantages of Ag detection assays
Only for a limited number of organisms
- Common ones → Giardia, Cryptosporidium, Entamoeba
For microscopic examination of wet mounts of preserved feces, what are usually destroyed and what remains intact?
Trophs are destroyed, cysts remain
An important component of light microscopes used in parasitology labs
Ocular micrometer
When making wet mounts, you should make two different types of mounts on the same slide…what are they?
Saline and iodine mounts
Smears of preserved, concentrated stool are made on a slide. Many modern fixatives permit specimen adherence to the slide. How do you make this slide?
Use an applicator stick (up and down motion) to make thick and thin areas
Dried smears are usually stained w/ what?
Trichrome stain
Stained smears should be first examined on low power and then use the ____ objective
100x oil immersion objective (1000x total magnification)
Three other methods besides trichrome to stain smears?
- Modified acid-fast stain (Cryptosporidium spp and coccida)
- Modified safranin stain (Cyclospora cayetanensis)
- Chromotrope R2/modified trichome (microsporidia)
Setback to molecular methods like the nucleic acid amplification test
Currently only available for detection of a limited number of intestinal protozoa
Enteric amoeba that lyses tissue
Entamoeba histolytica
Entamoeba histolytica
- Intestinal and extra-intestinal diseases
- Amoebic dysentery
- Amoebic colitis
- Amoebic abscesses
Entamoeba histolytica
- Geographic distribution
Worldwide but has a higher incidence of infection in developing countries
Entamoeba histolytica
- What disease does it cause?
Amoebiasis
Entamoeba histolytica
- Who are the hosts?
Humans are the only hosts
Entamoeba histolytica
- Infectious form
Cysts
Entamoeba histolytica
- How are cysts transmitted?
Fecal-oral route
Entamoeba histolytica
- Common settings
?
Entamoeba histolytica
- Test of choice?
EIA - high sensitivity and specificity
Entamoeba histolytica
- Must be distinguished from ____ ____ and other protozoa and also must be distinguished from ____ ____!!!
- Nonpathogenic amoebae
- Polymorphonuclear neutrophils
Entamoeba histolytica
- Size of cysts
12-15µm
Entamoeba histolytica
- Size of trophs
15-20µm
Entamoeba histolytica
- Is morphologically indistinguishable from what two organisms?
- Entamoeba dispar
- Entamoeba moshkovskii
How do you differentiate Entamoeba histolytica from Entamoeba hartmanni?
Exactly the same just much smaller
How do you differentiate Entamoeba histolytica trophs from Entamoeba coli trophs?
- E. histolytica: Karyosome in the center, is compact, peripheral chromatin is smoothly and evenly arranged “clean” cytoplasm
- E. coli: karyosome is off-center (messy looking), spread out, peripheral chromatin is clumpy and unevenly arranged in “dirty” cytoplasm
How do you differentiate Entamoeba histolytica cysts from Entamoeba coli cysts?
- E. histolytica: Nuclei usually paired on each side in 2 different focal planes, chromatic bodies with rounded ends
- E. coli: Nuclei scattered randomly throughout cyst in many different focal planes, chromatoid bodies with splintered ends
Only known pathogenic ciliate of humans
Balantidium coli
Balantidium coli
- Geographic distribution
Worldwide
Balantidium coli
- What disease does it cause?
Balantidiasis
Balantidium coli
- What are the hosts?
- PIGS (infections are more common in areas where pigs are raised)
- Rodents
- Primates
Balantidium coli
- Infectious form
Cyst
Balantidium coli
- Symptoms
- Most cases are asymptomatic
- Persistent diarrhea, dysentery, and abdominal pain
- Weight loss
- Can be severe in debilitated hosts
Balantidium coli
- What do we detect in the stool?
Trophs, not cysts
Balantidium coli
- Appearance of trophs
- CILIATED
- Big cell (40-200µm)
- Bean-shaped macronucleus
- Cytosome at narrower end
What is one of the most common intestinal parasites in the world?
Giardia intestinalis
Giardia intestinalis
- Other names
- G. lamblia
- G. duodenalis
Giardia intestinalis
- What disease does it cause?
Giardiasis
Giardia intestinalis
- Geographic distribution
Worldwide but is more prevalent in warm climates and in children (day care)
Giardia intestinalis
- How is it transmitted?
- Fecal-oral route
- Food
- Person-to-person
- Fomites (inanimate objects)
- Waterborne (most big outbreaks)
Giardia intestinalis
- Who are the hosts?
Humans and animals
Giardia intestinalis
- Common settings
- Children in daycares and their close contacts
- Campers and backpackers (drinking contaminated water)
Giardia intestinalis
- Infectious form
Cyst
Giardia intestinalis
- Symptoms
- Diarrhea, bloating, nausea, vomiting
- Foul-smelling feces and flatulence (purple burps) due to waste products from G. intestinalis
- Weight loss
- Dehydration
Giardia intestinalis
- Test of choice?
EIA - high sensitivity and specificity
Giardia intestinalis
- How often are the parasites in the feces?
Varies from day to day (get 3 samples over a period of 10 days)
Giardia intestinalis
- What motility should you look for in a fresh specimen?
“Falling leaf” motility
Giardia intestinalis
- Troph appearance
- External flagella, pear shape
- Two nuclei (look like eyes) sucking disc, median bodies
- 12µm
Giardia intestinalis
- Cyst appearance
- Small oval cyst
- 2-4 nuclei at one end
- Median bodies across middle, fibrils down the center
- 10µm
What is “fragile, enteric ‘amoeba’ w/ 2 nuclei” but is NOT an amoeba?
Dientamoeba fragilis
Dientamoeba fragilis
- What disease does it cause?
Dientameobiasis
Dientamoeba fragilis
- Geographic distribution
Worldwide
Dientamoeba fragilis
- How is it transmitted?
Presumably fecal-oral route
Dientamoeba fragilis
- Infectious form
Troph
Dientamoeba fragilis
- Appearance of troph
- WITHOUT external flagella
- Irregular rounded shape
- 2 nuclei
- “Dirty” cytoplasm
- 4-12µm
Dientamoeba fragilis
- Appearance of cyst
NO CYST FORM
Another name for pathogenic apicomplexa
Sporozoa
How do pathogenic apicomplex infect?
Invade and multiply w/in epithelial cells
Second most commonly diagnosed intestinal parasites in the U.S.?
Cryptosporidium spp
Cryptosporidium spp
- Geographic distribution
Worldwide
Cryptosporidium spp
- Habitat
- Waterborne → big outbreaks in Milwaukee, WI
Cryptosporidium spp
- Host
Humans and many vertebrate animals
Cryptosporidium spp
- How is it transmitted?
- Fecal-oral route
- Food
- Person-to-person
Cryptosporidium spp
- Four common settings
- Municipal water supply-associated outbreaks (Milwaukee)
- Children in daycare
- People in contact w/ farm animals
- People w/ AIDS who have unrelenting, water diarrhea
Cryptosporidium spp
- Infectious form
Oocyst
Cryptosporidium spp
- Significant symptom
Profuse watery diarrhea (up to 10L/day)
Cryptosporidium spp
- Test of choice
EIA - highest sensitivity and specificity
Cryptosporidium spp
- What color do they stain in a modified-Kinyoun stain?
Red b/c they’re ACID-FAST
Cryptosporidium spp
- Oocyst appearance
- 4-6µm
- 4 crescent-shaped sporozoite present w/ them
“Circular sporocytes”; a coccidian
Cyclospora cayetanensis
Cyclospora cayetanensis
- Geographic distribution
- Most common in tropical and subtropical regions, exported to other regions of the planet
- No consistent seasonal pattern of occurrence in endemic areas
Cyclospora cayetanensis
- What disease does it cause?
Cyclosporiasis
Cyclospora cayetanensis
- Who are the hosts?
Humans
Cyclospora cayetanensis
- How is it transmitted?
Fecal-oral route mainly through food and water, NOT DIRECT CONTACT
- Raspberries, basil, cilantro, other fresh produce
Cyclospora cayetanensis
- Infectious form
Sporulated oocyst
Cyclospora cayetanensis
- Highlights of the life cycle
- People ingest sporulated (mature) oocysts
- Unsporulated oocysts (in fresh feces) are not infectious
- Oocysts must mature in the environment for several days to weeks before they’re infectious
Cyclospora cayetanensis
- Symptoms
- Water diarrhea can be severe
- Weight loss, abdominal pain, nausea, vomiting, etc.
- Untreated infections can last a while (10-12 weeks)
- Dehydration, which is sometimes severe
- Relapse is not uncommon if initially untreated
Cyclospora cayetanensis
- Can be shed ____ so get more than one stool specimen
Intermittent
Cyclospora cayetanensis
- Wet mounts are useless, but oocysts are ____ and ____ when excited w/ certain wavelengths of light
- Acid-fast
- Autofluoresce
Cyclospora cayetanensis
- Size of unsporulated oocysts
8-10µm → bigger than Cryptosporidium (4-6µm)
Cyclospora cayetanensis
- Oocyst appearance
Small coccidian parasite, round, acid fast (can have ghost cells in af); stain red; no sports, wrinkled and crinkled
Cystoisospora belli
- Causes what disease?
Cystoisosporiasis
Cystoisospora belli
- Geographic distribution
Worldwide, especially in tropical and subtropical regions
Least common of the coccidia
Cystoisospora belli
Cystoisospora belli
- Who is the host?
Immunocompromised patients, especially those w/ AIDS
Cystoisospora belli
- How is it transmitted?
Fecal-oral route
Cystoisospora belli
- Significant symptom
Eosinophilia
Cystoisospora belli
- Infectious form
Mature oocyst
Cystoisospora belli
- It will ____ when excited w/ UV light
Autofluoresce
Cystoisospora belli
- Oocyst appearance
Large, ellipsoidal, contains 1 sporoblast (pig in a blanket)
Sarcocystis hominis and suihominis
- Causes what disease?
Sarcocystosis
Sarcocystis hominis and suihominis
- Geographic distribution
Worldwide; more common where livestock are raised
Sarcocystis hominis and suihominis
- Who are the hosts?
Humans are a dead-end host
Sarcocystis hominis and suihominis
- How is it transmitted?
By consumption of undercooked/contaminated meat
Sarcocystis hominis and suihominis
- Infectious form
??Oocyst??
Sarcocystis hominis and suihominis
- Symptoms
- Often asymptomatic but occassionally a mild fever, diarrhea, chills, and vomiting
Sarcocystis hominis and suihominis
- Oocyst appearance
Paired (shrink-wrapped) ovals
Sarcocystis hominis and suihominis
- Do these organisms autofluoresce?
YES
These are opportunistic, obligate, intracellular pathogens
Microsporidia
If microsporidia aren’t protozoa, what are they?
Fungi
Two types of microsporidia that are common human gastrointestinal pathogens
- Enterocytozoon bieneus
- Encephalitozoon intestinalis
Microsporidia
- Geographic distribution
Worldwide
Microsporidia
- Who are the hosts?
Severely immunocompromised AIDS patients
Microsporidia
- How is it transmitted?
Consumption of contaminated food and water, contaminated fomites, and possibly airborne?
Microsporidia
- Infectious form
Spore
Microsporidia
- Symptoms
Diarrhea
Microsporidia
- What stains should be used for the microscopic exam?
- Chromotrope 2R
- Ryan’s modified-trichrome stain
Microsporidia
- Gold test
- Test of choice?
- Gold test: transmission electron microscopy
- Choice: immunofluorescence
Encephalitozoon spp
- Appearance
Gram stain → resembles large GPRs
Blastocystis spp
- Infectious form
Cyst (cyst released also)
Blastocystis spp
- Role in disease
Debateable role (sometimes no symptoms, other times diarrhea etc.)
Chilomastix mesnili
- Pathogenic or nonpathogenic?
Nonpathogenic protozoa
Chilomastix mesnili
- Appearance of trophs
External flagella, lopsided shortened pear shape, one nucleus, cytosome looks like mouth (smiling fish)
Chilomastix mesnili
- Appearance of cysts
Small lemon shape cyst, one nucleus, cytosine small, looks like a shephard’s crook
Endolimax nana
- Pathogenic or nonpathogenic?
Nonpathogenic
Endolimax nana
- Appearance of troph
W/o peripheral nuclear chromatin, large irregular karyosome (nana with dilated pupil)
Endolimax nana
- Appearance of cyst
four nuclei (nana has 4 eyes), short oval shape, small nuclei surrounded by clear space
Enteromonas hominis
- Pathogenic or nonpathogenic?
Nonpathogenic
Enteromonas hominis
- Appearance
?
Iodamoeba butschlii
- Pathogenic or nonpathogenic?
Nonpathogenic
Iodamoeba butschlii:
- Cyst appearance
- Troph appearance
Cyst: large vacuole in the cyst
Pentatrichomonas hominis
- Pathogenic or nonpathogenic?
Nonpathogenic
Pentatrichomonas hominis
- Appearance of trophs
External flagella, lopsided pear shape, one nucleus, undulating membrane and axostyle (fish with spike)
Advantages of Ag detection methods
High sensitivity and specificity
Disadvantages of microscopic examination methods
- Objective analyses b/w techs
- Need baseline knowledge to ID
- Requires time to perform analyses
Advantages of microscopic examination methods
- Microscopes relatively inexpensive
- Can be purchase cordless
- Easy to train on how to use
- Don’t “expire”
Advantages of molecular methods
- Detectable parasites can be ID’d by the same assay as bacteria
- High sensitivity and specificity
Disadvantages of molecular methods
- Detrct either dead or alive parasites
- $$$