Lecture 2 - Protozoa Lumen Dwelling Amoeba
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1
What is a protozoa?
single-celled, eukaryotic
2
How are different types of protozoa (parasitic to humans) classified as?
Principle means of locomotion (amoeba, flagellates, ciliates)
3
What are the different types of protozoans which are parasitic to humans? (list 3)
Amoeba
Flagellates
Cilliates
Flagellates
Cilliates
4
How do amoebas move?
pseudopodia
5
How do flagellates move?
flagella
6
How do ciliates move?
cilia
7
How do protozoas reproduce?
asexually by fission. each parent gives 2 progeny.
8
How often do amoeba's reproduce?
randomly.
9
How do flagellates reproduce? in what way?
longitudinal
10
How do ciliates reproduce?
Transversely.
11
What is another name for amoebae?
Sarcodina.
12
What are the 3 main things to tell amoeba apart microscopically?
1. Nucleus/nuclei
2. Cellular inclusions
3. Trophoziote vs cyst morphology
2. Cellular inclusions
3. Trophoziote vs cyst morphology
13
what are things to look at in the nucleus of an amoeba?
number
distribution/concentration of chromatin
endosomes/karyosomes within the nucleoplasm <- size and alignment
distribution/concentration of chromatin
endosomes/karyosomes within the nucleoplasm <- size and alignment
14
What are things to look at in the cellular inclusions?
Vacuoles
RBCs <- usually indicates it is invasive/pathogenic
RBCs <- usually indicates it is invasive/pathogenic
15
If RBCs are within a parasite, what does that indicate?
invasive/pathogenic
16
What can the vacuoles in parasites contain?
glycogen
17
What is a trophozoite?
active and eating stage of a parasite.
18
What happens when a trophozoite in ingested?
Nothing. stomach low pH kills it.
19
What is a cyst?
closed-sac that protects the material inside from unfavorable external environment. dormant and resistant to the enviroment
20
How long can a cyst survive?
days/weeks
21
What happens in a cyst?
nuclear division
infective stage.
infective stage.
22
what is lumen dwelling?
commonly found in the alimentary tracts (GI tract) of their hosts
23
List 6 lumen dwelling amoebas
Entamoeba histolytica
Entamoeba hartmani
Entamoeba coli
Endolimax nana
Iodamoeba butshlii
Blastocyctis hominis
Entamoeba hartmani
Entamoeba coli
Endolimax nana
Iodamoeba butshlii
Blastocyctis hominis
24
What are the dimensions for trophozoites from Entamoeba histolytica?
15-20 um
25
What are the dimensions for cysts from Entamoeba histolytica?
12-15 um
26
How do the karyosomes (or endosomes) appear in the trophozoites for E. histolytica?
small and central (or slightly eccentric)
27
What does the nuclear chromatin look like in the trophozoites for E. histolytica?
fine, evenly distributed along nuclear membrane
28
How many nuclei can cysts from E. histolytica have?
up to 4 nuclei!
29
What other characteristic can a cyst from E. histolytica contain?
Chromatoid bodies with rounded ends
30
Is this a trophozoite or cyst?
trophozoite
31
Is this a trophozoite or cyst?
cyst
32
What is (A)?
chromatoid bodies
33
What type of cyst is this?????
mononucleated cyst
34
What type of cyst is this?
Binucleated cyst
35
If there are RBCs within a trophozoite, what does that mean?
Diagnostic for E. histolytica. Indicates it is an invasive species. 
36
Entamoeba histolytica can appear very similarly to
E. dispar
E. bangladeshi
E. moshkovskii
E. bangladeshi
E. moshkovskii
37
How is E. histolytica distinguished from other similar parasites?
PCR
38
What is the treatment for E. histolytica?
Relief of symptoms: Blood and fluid replacement - anemic patients
Metronidazole (Flagyl) w/ antibiotics
Metronidazole (Flagyl) w/ antibiotics
39
Where are E. histolytica cysts typically found?
FORMED stool
40
How can someone get E. histolytica?
exposed to feces via contaminated water/food/hands/sex
41
Where are trophozoites from E. histolytica typically found? (WHAT TYPE OF SAMPLE)
diarrheal stool
42
Where does excystation occur in the E. histolytica life cycle?
small intestine
43
What is excystation?
the process by which an organism escapes from a cyst or cyst wall
44
After excystation of E. histolytica, what happens?
trophozoites travel to large intestine.
45
What are the 3 different outcomes for E. histolytica?
A. remain confined to intestinal lumen = noninvasive infection
B. invade intestinal mucosa = intestinal disease (painful; lesions)
C. invade the blood reaching liver, brain , and lungs = extraintestinal disease
B. invade intestinal mucosa = intestinal disease (painful; lesions)
C. invade the blood reaching liver, brain , and lungs = extraintestinal disease
46
What are the 5 different clinical features that E. histolytica can present?
1. Intestinal infection
2. Hepatic infection
3. Brain abcess
4. Lung infection
5. asymptomatic/ non-invasive.
2. Hepatic infection
3. Brain abcess
4. Lung infection
5. asymptomatic/ non-invasive.
47
Which is the most common clinical feature for E. histolytica?
intestinal infection
48
Most worrisome clinical presentation in the intestine for E. histolytica is....
"Flask-like lesions/ uclers"
49
Why are "Flask-life lesions/ uclers" worrisome? (E. histolytica)
can result in perforation of the colon.
50
What is the most common extraintestinal presentation that E. histolytica can cause?
Hepatic infection
51
Which parasite caused this?
E. histolytica; hepatic infection containing 'anchovy paste'
52
What is an another name for an intestinal infection caused by E. histolytica?
Invasive amebiasis
53
What is amoebic dysentery?
an intestinal infection caused by the parasite Entamoeba histolytica.
54
What are the common symptoms seen intestinal infection caused by E. histolytica?
Diarrhea (intermittent), which can be bloody, cramping, flatulence.
55
What are symptoms seen in an hepatic infection caused by E. histolytica?
Fever, abdominal pain, jaundice, diarrhea (non-bloody), hiccups, loss of appetite, weight loss.
56
What is this ?
Brain abscess
57
Brain abscess caused by E. histolytica is usually seen also with
Hepatic infection (liver abscess)
58
What are symptoms seen in patients with brain abscess caused by E. histolytica?
Depends on area of brain affected
59
Lung infection caused by E.histolytica can lead to
penumonia, empyema (pus in pleural cavity)
60
What are some symptoms seen in lung infections caused by E. histolytica?
Fever, cough, shortness of breath, chest pain.
61
What is Entamoeba hartmani?
A non-pathogenic amoeba
62
What is this?
Entamoeba hartmani
63
How is E. hartmani transmitted and where does it reside?
Fecal-oral; lumen of colon/cecum
64
What is the main difference between E. histolytica vs. E. harmani?
E. hartmani (TROPHOZOITES) is smaller! &
will NOT see RBCs in cytoplasm bc its not invasive!
will NOT see RBCs in cytoplasm bc its not invasive!
65
What can you see in E. hartmani cysts?
1. 4 (mature) nuclei
(more common to see 1-2 <- immature)
(Looks like E. histolytica)
2. May see chromatin bodies
(more common to see 1-2 <- immature)
(Looks like E. histolytica)
2. May see chromatin bodies
66
What is the size of E. hartmani trophs/cysts?
5-10 um
67
What is the size of E. histolytica trophs/cysts?
12-20 um
68
What is an Entamoeba coli?
Nonpathogenic amoeba
69
Where in the world is Entamoeba coli found?
Worldwide
70
How is Entamoeba coli transmitted?
Fecal-oral; Colon/cecum lumen
71
What do the trophozoites of Entamoeba coli look like?
Large, relatively speaking 15-20 um
Nucleus has large, usually eccentric karyosome
Peripheral chromatin usually coarse, irregular granules
Nucleus has large, usually eccentric karyosome
Peripheral chromatin usually coarse, irregular granules
72
What do the cysts of Entamoeba coli look like?
Spherical -> oval shaped
Mature cysts = 8 nuclei
(Supernucleate cysts can have 16 or more!)
May contain diffuse glycogen
Chromatin bodies (splinter like)
Mature cysts = 8 nuclei
(Supernucleate cysts can have 16 or more!)
May contain diffuse glycogen
Chromatin bodies (splinter like)
73
What type of reagent is used to dye fresh poo?
Iodine
74
Label the structure, starting from top and then clockwise. 
1 Nucleus
Vacuoles
8 Nuclei
Cyst - stained with iodine (10-35 um)
Cytoplasmic granules
Trophozoite stained with Giemsa (15-50 um)
Vacuoles
8 Nuclei
Cyst - stained with iodine (10-35 um)
Cytoplasmic granules
Trophozoite stained with Giemsa (15-50 um)
75
What amoeba is it?
Entamoeba coli
(troph - purple; cyst - iodine)
(troph - purple; cyst - iodine)
76
Cysts or trophs?
Cysts
77
Cysts or trophs?
Trophs
78
Label the arrow
Spindle-shaped chromatin bodies
79
Label the arrow
E. coli is infected with fungus.
80
what are these? (which one is Entamoeba histolytica and which one is Entamoeba coli)
left: E.histolytica
right: E. coli
right: E. coli
81
Which one is Entamoeba histolytica and which one is Entamoeba coli?
top: Entamoeba histolytica
bottom: Entamoeba coli
bottom: Entamoeba coli
82
Is endolimax nana pathogenic or nonpathogenic?
Non pathogenic
83
Where can you find Endolimax nana? (location on earth)
worldwide
84
What type of transmission is Endolimax nana required for its cycle?
Fecal-oral
85
Where inside the body is Endolimax nana found?
Lumen of colon/cecum
86
Are the trophozoites of Endolimax nana big or small?
small
87
What is the main appearance of Endolimax nana that differentiate from other trophozoites?
Large, irregularly shaped karyosome
88
Does endolimax nana have peripheral chromatin?
NO, clear around the karyosome
89
Are there any inclusions in Endolimax nana?
Yes, the cytoplasm is highly vacuolated and coarsely granuar
90
What other organims is Endolimax nana mistaken with?
Iodamoeba butschlii
91
What is one key difference between Endolimax nana and Iodamoeba butschlii?
E. nana lacks granules surrounding karyosome
92
Are the cysts for Endolimax nana big or small?
Small
93
What shape are the cysts for Endolimax nana?
Spherical -> elliptical
94
How many nuclei do E. nana cysts have?
4
95
How many nuclei for Iodamoeba butschlii cysts have?
1 nucleus
96
What is one main difference in Endolimax nana cysts that differentiate it from the rest?
Large karyosome
97
Do E. nana cysts have chromatin?
No peripheral chromatin
98
What may E. nana cysts contain?
Diffuse glycogen and granules/inclusions
99
Do cysts of E. nana have chromatin bodies?
No
100
What is Iodamoeba butschlii?
Nonpathogenic organism
101
Where can you find Iodamoeba bustchlii in the world?
worldwide
102
Where in the body does Iodamoeba butschlii reside?
Lumen of colon/cecum
103
What is the mode of transmission for Iodamoeba buschlii?
Fecal-oral
104
Are the karysomes of Iodamoeba butschlii trophozoites large or small?
Large, centrally located
105
What can be frequently seen around the karyosome of Iodamoeba butschlii trophozoites?
Refractile granules
106
What does the cytoplasm of iodamoeba butschlii trophozoites look like?
Cytoplasm can be vacuolated, coarse, granular
107
What can Iodamoeba butschlii trophozoite be misidentified as?
E. nana
108
What are the shape of Iodamoeba bustchlii cysts look like?
Spherical -> ellipsoidal
109
How much nuclei does Iodamoeba butschlii mature cysts contain?
1 nucleus
110
Are the karysomes of Iodamoeba butschlii cycts large or small?
Large, (usually) eccentric karyosome
111
What is present in the cytoplasms of Iodamoeba butschlii?
Discreet, compact mass of glycogen in cytoplasm
112
Which nonpathogenic organism is this?
Iodamoeba butschlii
113
Is Blastocystis hominis common or rare?
Common and reportable - "blastocystis species"
114
Is blastocystis hominis found in symptomatic or asymptomatic patients?
Both
115
What is blastocystis hominis linked to?
Irritable bowel syndrome
116
Where can Blastocystis hominis be found in the world?
worldwide
117
What is the likely mode of transmission for Blastocystis hominis?
Fecal-oral
118
Where in the body does blastocystis hominis like to hang out?
Intestinal tract
119
What does the cyst-like stage of Blastocystis hominis look like?
Large central body (visibly resembling a vacuole)
Narrow rim of cytoplasm w/ nuclei and inclusion bodies
Possible infectious stage
Narrow rim of cytoplasm w/ nuclei and inclusion bodies
Possible infectious stage
120
What lab sample is blastocystis hominis seen in lab?
feces
121
What organims is this cyst from?
blastocystis hominis
122
Name the organism (left to right) 
Endolimax nana
Entamoeba hartmanni
Iodamoeba butschlli
Entamoeba polecki (DON'T NEED TO KNOW)
Entamoeba coli
Entamoeba histolytica/dispar
Entamoeba hartmanni
Iodamoeba butschlli
Entamoeba polecki (DON'T NEED TO KNOW)
Entamoeba coli
Entamoeba histolytica/dispar
123
What organism is this and how many nuclei does is have?
Endolimax nana; 4 nuclei
124
What organism is this and what are some key notes about it?
Entamoeba hartmanni
Small mature cysts
4 nuclei
GRAPE-LIKE CHROMATOID BODIES
Immature cysts have 1-2 nuclei
Small mature cysts
4 nuclei
GRAPE-LIKE CHROMATOID BODIES
Immature cysts have 1-2 nuclei
125
What organism is this and what are some key notes about it?
Iodamoeba butschlli
Single nucleus
Large karyosome
Large glycogen vacuole
Single nucleus
Large karyosome
Large glycogen vacuole
126
What organism is this and what are some key notes about it?
Entamoeba coli
8 nuclei
Chromatoid bodies with splintered ends
Immature cysts can have 1-4 nuclei and glycogen mass
8 nuclei
Chromatoid bodies with splintered ends
Immature cysts can have 1-4 nuclei and glycogen mass
127
What organism is this and what are some key notes about it?
Entamoeba histolytical/dispar
4 nuclei
Chromatoid bodies with rounded ends
Immature cysts can have 1-2 nuclei and glycogen mass
4 nuclei
Chromatoid bodies with rounded ends
Immature cysts can have 1-2 nuclei and glycogen mass
128
Name the organism (left to right) 
Entamoeba histolytica
Entamoeba hartmanni
Entamoeba coli
Entamoba polecki (do not need to know) - rare
Endolimax nana
Iodamoeba butschlii
Entamoeba hartmanni
Entamoeba coli
Entamoba polecki (do not need to know) - rare
Endolimax nana
Iodamoeba butschlii
129
What is the best diagnostic testing for trophozoites?
fresh diarrhea
130
What is the best diagnostic testing for lumen dwelling parasites?
Fresh stool
131
How to prepare a sample of stool?
fix sample with formalin or PVA in a conical tube and concentrat the specimen.
132
What does fixing do when preparing the specimen?
Helps maintain and remove debris
133
What are common stains for stool examination?
Iodine and trichrome
134
What is iodine good for ID?
Protozoa and cysts; which take up the dye and appear brown while other objects in the sample remain clear.
135
What colors does trichrom stain?
pink, blue, purple
136
What other diagnostic testing is used for stool samples besides fixing?
PCR and GI panels for parasite, virus, and bacteria
137
What type of tests are performed for specimens in 10% formalin?
Wet mount, ELISA, Chromotrope stain
138
What type of tests are performed for specimens in Formalin-ethyl acetate concentration?
Wet mount, direct mount, acid-fast stain, direct immunofluorescent assay, safranin stain
139
what type of tests are performed for specimens in PVA fixative?
Trichrome stain 
140
Wet mount with specimens in 10% formalin tests which organisms?
Helminths and protozoa
141
ELISA tests which organisms?
Giardia and Cryptosporidium
142
Chromotrope stain tests which organisms?
Microsporidia
143
Wet mount with formalin-ethyl acetate concentration tests which organisms?
Helminths and protozoa
144
Direct mount test which organisms?
epifluorescence for Cyclospore and Isospora
145
Acid fast stain tests which organisms?
Cryotosporidium, cyclospora, and isospora
146
Direct immunofluorescent assay tests which organisms?
Giardia and Cryptosporidium
147
Safranin stain tests which organisms?
Cyclospora
148
Trichrome stain tests which organism?
Protozoa
149
What are Antiamoebic agents?
drugs that treat amoebiasis, a protozoan infection caused by the parasite Entamoeba histolytica
150
What does antiamoebic agent focus treating which organism?
E. histolytica
151
What determines the preferred antiamoebic therapy?
Clinical presentation
152
What drug is used for luminal clinical presentation of E. histolytica?
paromomycin
153
What drug is used for tissue/systemic clinical presentation of E. histolytica?
metronidazole (flagyl) and tinidazole are useful for hepatic abscesses.