Exercise No. 6c-2 PROTOZOA Phylum Sarcomastigophora - Subphylum Mastigophora - Hemoflagellates Flashcards
(107 cards)
1
Q
four morphologic forms of clinical significance associated with hemoflagellates
A
amastigote, promastigote, epimastigote, and trypomastigote
2
Q
amastigote was formerly called
A
leishmania
3
Q
promastigote was formerly called
A
leptomonas
4
Q
epimastigote was formerly called
A
crithidia
5
Q
trypomastigote was formerly called
A
trypanosoma
6
Q
This stage is adapted to intracellular existence, being found inside macrophages, monocytes, neutrophils, or endothelial cells.
A
Amastigote
7
Q
It is also known as the L-D (Leishman-Donovan) body.
A
Amastigote
8
Q
The body is slightly wider than that of the promastigote.
A
Epimastigote
9
Q
Volutin granules are found in the cytoplasm.
A
Trypomastigote
10
Q
shape and size: ovoidal in shape and, on an average, measures 5 µm long by 3 µm wide
A
Amastigote
11
Q
shape and size: spindle-shaped and measures 9 to 15 µm in length
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Promastigote (typical)
12
Q
shape and size: spindle-shaped and measures approximately 9 to 15 µm in length
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Epimastigote (average)
13
Q
shape and size: may be long and slender or assumes the shape of the letters C, S or U, and measures 12 to 35 µm long by 2 to 4 µm wide
A
Trypomastigote
14
Q
nucleus: large single nucleus that is typically located off-center, sometimes present more toward the edge of the organism
A
Amastigote
15
Q
nucleus: large single nucleus is located in or near the center of the body
A
Promastigote
16
Q
nucleus: large single nucleus
A
Epimastigote; Trypomastigote
17
Q
kinetoplast: consists of a deeply staining rod-like parabasal body and adjacent dot-like blepharoplast
A
Amastigote
18
Q
kinetoplast: located in the anterior end of the organism and gives rise to an axoneme
A
Promastigote
19
Q
kinetoplast: located anterior to the nucleus
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Epimastigote
20
Q
kinetoplast: located posteriorly from which emerges a full body length undulating membrane
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Trypomastigote
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Q
blepharoplast: gives rise to and is attached to an axoneme
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Amastigote
22
Q
axoneme: extends to the anterior tip of the organism
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Amastigote
23
Q
flagellum: absent
A
Amastigote
24
Q
flagellum: single free; extends anteriorly from the axoneme
A
Promastigote
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undulating membrane: short; arising from the kinetoplast forms into a free flagellum at the anterior end
Epimastigote
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undulating membrane: short; arising from the kinetoplast forms into a free flagellum at the posterior end
Trypomastigote
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involve some combination of the four morphologic forms
Leishmania and Trypanosoma
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Leishmania and Trypanosoma species are morphologically indistinguishable, but they can be differentiated by
isoenzyme analysis, molecular methods, or monoclonal antibodies
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move by means of flagella
hemoflagellates
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shape and size: ovoidal in shape and, on an average, measures 5 µm long by 3 µm wide
Amastigote
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shape and size: spindle-shaped and measures 9 to 15 µm in length
Promastigote (typical)
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shape and size: spindle-shaped and measures approximately 9 to 15 µm in length
Epimastigote (average)
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shape and size: may be long and slender or assumes the shape of the letters C, S or U, and measures 12 to 35 µm long by 2 to 4 µm wide
Trypomastigote
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nucleus: large single nucleus that is typically located off-center, sometimes present more toward the edge of the organism
Amastigote
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nucleus: large single nucleus is located in or near the center of the body
Promastigote
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nucleus: large single nucleus
Epimastigote; Trypomastigote
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kinetoplast: consists of a deeply staining rod-like parabasal body and adjacent dot-like blepharoplast
Amastigote
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kinetoplast: located in the anterior end of the organism and gives rise to an axoneme
Promastigote
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kinetoplast: located anterior to the nucleus
Epimastigote
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kinetoplast: located posteriorly from which emerges a full body length undulating membrane
Trypomastigote
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blepharoplast: gives rise to and is attached to an axoneme
Amastigote
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axoneme: extends to the anterior tip of the organism
Amastigote
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flagellum: absent
Amastigote
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flagellum: single free; extends anteriorly from the axoneme
Promastigote
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undulating membrane: short; arising from the kinetoplast forms into a free flagellum at the anterior end
Epimastigote
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The specimen of choice depends on the nature of disease.
Leishmania
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Fluid aspirate beneath the ulcer bed, biopsy of skin ulcer
Old World cutaneous leishmaniasis (CL)
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Same as in CL, and mucosal scrapings
Mucocutaneous leishmaniasis (ML)
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Tissue aspirates (liver, splenic, sternal marrow), blood (buffy coat)
Visceral leishmaniasis (VL)
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Stains used in Direct microscopy of Leishmania
Giemsa, Wright's or Leishman's stains, or Hematoxylin-Eosin
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Direct microscopy Objective: Leishmania
oil immersion objective
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In culture, Leishmania species behave as if they are in the
insect vector
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NNN medium was described by
Novy, MacNeal and Nicolle
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This is a rabbit blood agar slope consisting of two parts of salt agar and one part of defibrinated rabbit blood.
NNN medium
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NNN medium: The material is inoculated into the water of condensation and culture is incubated at
22- 24°C for 1-4 weeks.
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NNN medium: At the end of each week, a drop of culture fluid is examined under
high power objective or phase contrast illumination
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This medium was originally designed for cultivation of insect tissue culture cells.
Schneider’s Drosophila medium
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This medium is used containing inactivated fetal bovine serum and antibiotic and antimycotic solution to isolate and grow etiologic agents of leishmaniasis.
Schneider’s Drosophila medium
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Immunodiagnosis: Serologic testing for Leishmania
IFA (indirect fluorescent antibody), ELISA (enzyme-linked immunosorbent assay), and DAT (direct agglutination test)
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Molecular diagnosis: Leishmania
Western blot and PCR
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confined to specialized laboratories and is yet to be used for routine diagnosis of visceral leishmaniasis in endemic areas
PCR
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delayed hypersensitivity test
Leishmanin skin test (Montenegro test)
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Skin test: What is injected intradermally on the dorsoventral aspect of forearm?
0.1 mL of killed promastigote suspension (106 washed promastigotes/mL)
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Leishmanin skin test (Montenegro test) Positive result
induration and erythema of 5 mm or more after 48-72 hours
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Leishmanin skin test (Montenegro test)
It has been tested in all types of leishmaniasis with high percentage of positivity in cutaneous leishmaniasis and 95% positivity in mucocutaneous leishmaniasis.
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Leishmanin skin test (Montenegro test)
In active visceral leishmaniasis, this test is negative and becomes positive usually 6-8 weeks after cure from the disease.
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The concentration of gammaglobulins in blood increases considerably following infection with
L. donovani
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In visceral leishmaniasis, the test becomes positive only when infection is at least three months old and may remain so even after six months of cure.
Nonspecific serum test
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1 mL of clear serum from the patient is taken in a small test tube, a drop of formalin (40% formaldehyde) is added, shaken and kept in a rack at room temperature. A control tube with normal serum is also set up.
walang answer nakalagay, no. 24 sa may "laboratory" part ng exercise 6c2
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Aldehyde test of Napier (or formol gel test): specimen
1 mL of clear serum
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Aldehyde test of Napier (or formol gel test): reagent
1 drop of formalin (40% formaldehyde)
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Aldehyde test of Napier (or formol gel test): incubation
room temperature
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A control tube with normal serum is also set up.
Aldehyde test of Napier (or formol gel test)
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Aldehyde test of Napier (or formol gel test): positive result
opaque jellification of the test serum
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Aldehyde test of Napier (or formol gel test): positive result resemblance
coagulated white of egg
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Aldehyde test of Napier (or formol gel test): positive result (reaction time)
3-30 minutes
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Antimony test of Chopra: specimen
0.2 mL of serum diluted 1:10 with distilled water
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Antimony test of Chopra: reagent
4% solution of urea stibamine
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Aldehyde test of Napier (or formol gel test): glassware
small ordinary test tube
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Antimony test of Chopra: glassware
Dreyer's tube
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Antimony test of Chopra: positive result
flocculent precipitate
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microscopic examination of chancre fluid, lymph node aspirates, blood, bone marrow, or, in the late stages of infection, cerebrospinal fluid demostrates
highly pleomorphic trypanosomes
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Concentration techniques such as centrifugation can be used prior to microscopic examination.
Trypanosoma
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Trypomastigotes are barely visible, but their motion against the RBCs makes them apparent.
Wet mount preparation
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The specimen is fixed on a slide and stained with Giemsa (most sensitive) or other Romanowsky’s stain (Wright, Leishman)
Permanent staining
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malaria diagnosis
Quantitative buffy coat (QBC) technique
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Almost all patients with African trypanosomiasis have very high levels of which immunoglobulin classes?
IgM and IgG
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Immunodiagnosis: Serologic testing for Trypanosoma
Indirect hemagglutination (IHA), indirect immunofluorescence (lF), enzyme-linked immunosorbent assay (ELISA), card agglutination trypanosomiasis test (CATT), and complement fixation test (CFT)
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Immunodiagnosis: Antigen detectionfor Trypanosoma
ELISA
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T or F: Molecular diagnosis: Nucleic acid-based tests are validated for the diagnosis of African trypanosomiasis, and diagnostic performance varies considerably among these tests.
F - No nucleic acid-based tests are validated for the diagnosis
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Direct microscopy: Trpanosoma acute stage specimen
trypomastigotes in circulating blood or cerebrospinal fluid (CSF)
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Trypomastigotes are barely visible, but their motion against the RBCs makes them apparent.
Wet mount preparation
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Trypomastigotes can be seen in thick and thin blood smear stained with Giemsa (most sensitive) or other Romanowsky’s stain (Wright, Leishman)
Permanent staining
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malaria diagnosis
Quantitative buffy coat (QBC) technique
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Direct microscopy: Trpanosoma chronic stage specimen
biopsy specimens stained with hematoxylin-and-eosin (H&E) or Giemsa
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also used for growing T. cruzi
NNN (Novy, MacNeal and Nicolle) medium
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found in trypanosoma culture
Epimastigotes and trypomastigotes
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This is the method of choice in suspected Chagas' disease, if other examinations are negative, especially during the early phase of the disease onset.
Xenodiagnosis
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allowed to feed on patient suspected to have Chagas’ disease
pathogen-free laboratory-bred triatomine bugs
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The bugs' feces are examined 3-4 weeks later for the presence of trypomastigotes.
Xenodiagnosis
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a useful technique for determining whether the patient is infected
Immunodiagnosis
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parasitemia is low
chronic phase of infection
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can be detected in urine and sera in patients with chronic Chagas disease
T. cruzi antigen
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Other tests: Trypanosoma
Electrocardiography (ECG) and chest X-ray; Endoscopy
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useful for diagnosis and prognosis of cardiomyopathy seen in chronic Chagas disease
Electrocardiography (ECG) and chest X-ray
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helps in visualization of megaesophagus in Chagas disease
Endoscopy
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Which test/tests give false-positive reactions in several other disease such as multiple myeloma, cirrhosis of liver, tuberculosis, leprosy, shictosomiases , African trypanosomiasis, etc. where hypergammaglobulinemia is a feature?
Nonspecific serum test (Aldehyde test of Napier and Antimony test of Chopra)
