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Parasitology-Lab > U3 - Diagnostic Parasitology > Flashcards

U3 - Diagnostic Parasitology Flashcards

1
Q

common reagents

A
  1. carbon fuchsin
  2. formalin (5% and 10%)
  3. lugol’s iodine solution
  4. SAF (sodium acetate-acetic acid formalin)
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2
Q

most common fixative

A

formalin

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3
Q

T/F:
All stains favor formalin.

A

False;
not all

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4
Q

2 types of stool analysis

A

macroscopic
microscopic

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5
Q

gross examination of stool

A

macroscopic analysis

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6
Q

analysis of stool under a microscope

A

microscopic analyses

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7
Q

factors analyzed macroscopically

A
  1. consistency
  2. color
  3. presence of blood
  4. presence of mucus
  5. presence of adult worm
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8
Q

type of stool examined macroscopically

A

fresh stool

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9
Q

simplest and easiest technique to facilitate detection of intestinal parasites that infected subjects pass in their feces

A

direct fecal smear

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10
Q

presence of intestinal protozoa (trophozoites or cysts)
or helminth eggs can be observed directly with what type of microscope?

A

light microscope

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11
Q

Small amount of fresh feces is mixed with either __________ or ___________ ____________.

A

saline
lugol/ iodine solution

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12
Q

purpose of saline

A

to detect the protozoa motility

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13
Q

purpose of lugol/iodine solution

A

to reveal the parasite structure

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14
Q

What does NSS stand for?

A

Normal Saline Solution

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15
Q

concentration of NSS

A

0.85% NaCl

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16
Q

type of stool used for direct wet preparation

A

fresh stool

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17
Q

useful technique if parasite is in low count

A

concentration

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18
Q

most ideal technique

A

permanent stain

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19
Q

type of stool used for permanent stain

A

fresh or preserved

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20
Q

T/F:
In permanent stain technique, stain must match the fixative.

A

True

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21
Q

NSS observation

A

morphology and motility

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22
Q

Lugol’s iodine observation

A

morphology
(cannot observe motility)

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23
Q

Organism killed by lugol’s iodine

A

trophozoites

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24
Q

organisms observed by saline

A
  1. motile trophozoites and larvae
  2. red blood cells
  3. Leukocytes
  4. Charcot–Leyden crystals
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25
Q

organisms observed by iodine solution

A

cysts of protozoa

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26
Q

When examining diarrheic or liquid feces containing mucus, which preparation should be applied to the mucous part of the stools?

A

both saline and iodine solutions

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27
Q

Explain the procedure of preparation.

A
  1. left side: 1 drop of saline; right side: 1 drop of lugol’s iodine
  2. 2 mg of fecal specimen emulsified in saline drop
  3. do the same with iodine drop
  4. place a cover slip on each suspension
  5. examine with a microscope
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28
Q

consideration in emulsifying stool in 2 solutions

A

separate applicator sticks must be used

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29
Q

chemical that kills any organisms present; thus, no motility is seen

A

iodine

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30
Q

specific procedure in placing coverslips

A
  1. place on each suspension touching the edge of the drop (angled)
  2. gently lower the coverslip onto the slide
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31
Q

purpose of angular method of coverslip upon placement

A

so no air bubbles are produced

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32
Q

recommended for monitoring large-scale treatment programs implemented for the control of soil-transmitted helminth infections

A

Kato-Katz Technique

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33
Q

Why is kato-katz technique the most recommended method for STH control?

A
  1. simple format
  2. ease of use in the field
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34
Q

What type of technique is kato-katz?

A

quantitative technique
(egg-counting)

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35
Q

purpose of kato-katz technique

A

quantifying eggs and establishing burden of intestinal infection

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36
Q

kato-katz technique
manner of reporting

A

eggs per gram stool (epg)

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37
Q

another purpose of kato-katz technique

A

to assess the efficacy of a treatment (if the number of eggs were reduced)

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38
Q

correlate the severity of the disease with the intensity of the infection

A

kato-katz technique

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39
Q

T/F:
kato-katz technique is applicable to all parasites

A

False;
not applicable to all;
some cases: no matter the amount, disease severity is the same

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40
Q

helminths that need soil to develop

A

soil-transmitted helminthes

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41
Q

causes of acquiring STH

A

poor sanitary and hygiene practices

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42
Q

unholy trinity according to WHO

A
  1. Trichuris trichiuria
  2. Ascaris lumbricoides
  3. Hookworm (Necator americanus and Ancylostoma duodenale)
    *** also Schistosoma spp.
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43
Q

materials for kato-katz technique

A
  1. wooden applicator sticks
  2. screen
  3. template
  4. spatula
  5. microscope slides
  6. hydrophilic cellophane
  7. jar, forceps, toilet paper, tissue
  8. newspaper
  9. glycerol-malachite green
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44
Q

glycerol-malachite green solution is poured into ____________ _________ in a jar and left for at least ___ hours prior to use.

A

cellophane strips
24

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45
Q

clearing agent

A

glycerine/ glycerol

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46
Q

Kato-katz technique procedure

A
  1. prepare materials
  2. place sample on a newspaper
  3. press nylon screen on top
  4. scrape the sieved fecal material from screen
  5. label glass slide
  6. place template with hole on center of slide
  7. fill hole in template with sired fecal material (avoid air bubbles and level feces off)
  8. lift off template, place in a bucket of water with concentrated detergent and disinfectant for reuse
  9. place cellophane (soaked in glycerol solution overnight) over fecal sample
  10. invert microscope slide
  11. press sample against cellophane strip on another slide or surface to spread feces in circle
  12. pick up slide (avoid cellophane separation)
  13. place it on bench with cellophane upwards
  14. water evaporates while glycerol clears feces
  15. when clarified, newspaper text must be read
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47
Q

Keep the slide for ___ hour(s) at ______ temperature to clear the fecal material prior to examination under the microscope.

A

1
room

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48
Q

Keeping the slide for an hour does not apply to __________.

A

hookworms
(eggs - thin shells)

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49
Q

Why must it not exceed 1 hour

A

might overclear and destroy organisms

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50
Q

done to speed up clearing and examination and allow action of the clearing agent

A

slide is placed in incubator
or kept in direct sunlight for several minutes

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51
Q

incubator temp

A

40 C

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52
Q

STH visibility/ recognizability duration

A

Ascaris lumbricoides: many months
Trichuris trichuria eggs: many months
Hookworm eggs: 30-60 minutes
Schistome eggs: several months (but examination within 24 hrs is preferred)

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53
Q

clear rapidly

A

hookworm eggs

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54
Q

In kato-katz technique, the smear should be examined _____________.

A

systematically

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55
Q

To give the number of eggs per gram of feces, what should the computation be?

A

egg count x factor (depending on template)

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56
Q

corresponding factors of used templates

A

50 mg template: x20
20 mg template: x50
41.7 mg template: x24

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57
Q

corresponding factor for 50 mg template

A

x20

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58
Q

corresponding factor for 20 mg template

A

x50

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59
Q

corresponding factor for 41.7 mg template

A

x24

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60
Q

template dimensions for amounts of stool

A

50 mg (x20): D=9mm; T=1mm
20 mg (x50): D=6.5mm; T=0.5mm
41.7 mg (x24): D=6mm; T=1.5mm

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61
Q

template dimensions for 50 mg stool

A

D=9mm; T=1mm

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62
Q

template dimensions for 20 mg stool

A

D=6.5mm; T=0.5mm

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63
Q

template dimensions for 41.7 mg stool

A

D=6mm; T=1.5mm

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64
Q

classification of intensity of infection for:
Ascaris lumbricoides
Trichuris trichuria
Hookworms
Schistosoma mansoni

A

Ascaris lumbricoides:
light: 1-4,999 epg
moderate: 5,000-49,000 epg
heavy: ≥50,000 epg

Trichuris trichuria:
light: 1-999 epg
moderate: 1,000-9,999 epg
heavy: ≥10,000 epg

Hookworms:
light: 1-1,999 epg
moderate: 2,000-3,999 epg
heavy: ≥4,000 epg

Schistosoma mansoni:
light: 1-99 epg
moderate: 100-299 epg
heavy: ≥400 epg

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65
Q

classification of intensity of infection for:
Ascaris lumbricoides

A

light: 1-4,999 epg
moderate: 5,000-49,000 epg
heavy: ≥50,000 epg

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66
Q

classification of intensity of infection for:
Trichuris trichuria

A

light: 1-999 epg
moderate: 1,000-9,999 epg
heavy: ≥10,000 epg

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67
Q

classification of intensity of infection for:
Hookworms

A

light: 1-1,999 epg
moderate: 2,000-3,999 epg
heavy: ≥4,000 epg

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68
Q

classification of intensity of infection for:
Schistosoma mansoni

A

light: 1-99 epg
moderate: 100-299 epg
heavy: ≥400 epg

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69
Q

2 types of blood film for malaria parasites

A

thick blood smear
thin blood smear

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70
Q

blood film used to determine if parasite is present

A

thick blood smear

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71
Q

blood film used to confirm the Plasmodium species present

A

thin blood smear

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72
Q

thick blood smear purpose

A

screening purposes

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73
Q

thin blood smear purpsoe

A

confirmatory purposes

74
Q

3 examples of blood parasites and disease caused

A
  1. plasmodium - malaria
  2. ticks - babesia
  3. hemoflagellates - trypanosomes
75
Q

type of blood specimen collected for thick and thin smear

A

capillary
venous

76
Q

more preferred blood specimen for thick and thin smear

A

capillary; directly observed without anticoagulant

77
Q

Why is venous blood less preferred for malarial examination?

A

stored in EDTA - may alter parasite structure

78
Q

gold standard

A

thick and thin blood smears

79
Q

smear that is performed first

A

thick blood smear

80
Q

stain preparation in thick blood smear for blood parasite

A

Giemsa stain

81
Q

giemsa stain pH

A

7.2

82
Q

blood smear diameter for thick smear

A

1-2 cm diameter

83
Q

rationale behind proper preparation technique for blood smears

A

to prevent parasites from obscuring behind the red blood cells

84
Q

stain preparation in thin blood smear for blood parasite

A

fixed with methanol
before Giemsa stain

85
Q

Why should one start examining blood parasites with thick smear?

A

You might miss out on the parasite if you don’t prioritize thick smear.

86
Q

Compare thick and thin films:
RBCs

A

Thick: lysed RBCs
Thin: fixed RBCs, single layer

87
Q

Compare thick and thin films:
volume

A

Thick: larger volume
Thin: smaller volume

88
Q

Compare thick and thin films:
blood volume/ fields

A

Thick: 0.25 ul blood/ 100 fields
Thin: 0.005 ul blood/ 100 fields

89
Q

Compare thick and thin films:

A

Thick:
Thin:

90
Q

Compare thick and thin films:
elements

A

Thick: more concentrated
Thin: good species

91
Q

Compare thick and thin films:
disadvantage

A

Thick: difficult species diagnosis
Thin: requires more time to read

92
Q

Compare thick and thin films:
density

A

Thick: parasite density
Thin: low density infections can be missed

93
Q

Compare thick and thin films:
result

A

Thick: positive or negative
Thin: parasite species

94
Q

template for thick and thin blood smear

A

Sub. ID
Intls.
TimePt.
Date
Time

95
Q

Explain blood smear procedure

A
  1. use separate slides
  2. thin film: hold spreader slide at 45-degree angle
  3. thin film: wait until blood spreads along edge
  4. thin film: push spreader slide forward
  5. thick film: spread drop using slide corner
  6. wait until both are dry; fix thin film with methanol
  7. if same slide: fix only thin film
  8. when both are dry, stain
96
Q

T/F:
thick smear should not be too thick

A

True;
must be able to read newspaper text through it

97
Q

methanol concentration for thin film fixation

A

100% (absolute) methanol

98
Q

How long does it take the thick smears to dry?

A

1-2 hours

99
Q

unstained blood smears must be protected from?

A

excessive heat
moisture
insects

100
Q

where should unstained blood smears be stored?

A

covered box

101
Q

how to quantify malaria parasites against RBCs

A

count the parasitized RBCs among 500-2,000 RBCs on the thin smear and express the results as %parasitemia

102
Q

% parasitemia formula

A

% parasitemia = (parasitized RBCs/total RBCs) × 100

103
Q

examination if parasitemia is:
high:
low:

A

high: [e.g., > 10%], examine 500 RBCs
low [e.g., <1%] ,examine 2,000 RBCs (or more)

104
Q

T/F:
Asexual blood stage parasites and gametocytes must be counted separately.

A

True

105
Q

T/F:
Only the former are clinically important and gametocytes of P. falciparum can persist after elimination of asexual stages by drug treatment.

A

True

106
Q

purpose of cellophane swab

A

for diagnosis of:
1. Enterobiasis caused by a nematode, Enterobius vermicularis
2. Taeniasis caused by tapeworm, Taenia solium or Taenia saginata

107
Q

other terms for cellophane swab

A

perianal swab
scotch tape swab
cellophane tape swab

108
Q

pinworm/ social worm causing enterobiasis

A

Enterobius vermicularis

109
Q

itching in perianal area

A

pruritus ani

110
Q

When do nematodes (Enterobius vermicularis) usually lay eggs in the perianal region?

A

at night
when moisty
when humid

111
Q

MOT for Enterobius vermicularis

A

fecal oral
inhalation

112
Q

a tapeworm that has radial segmentations

A

Taenia saginata

113
Q

shedding of gravid proglottids by the tapeworm

A

apolysis

114
Q

segments or units of the tapeworm’s body, often found in the perianal region or around the anus in cases of tapeworm infection

A

proglottids

115
Q

also known as pinworm infection

A

Enterobiasis

116
Q

Specimen of choice for the detection of Enterobius vermicularis (pinworm) eggs

A

cellophane swab

117
Q

Explain how eggs are secreted by parasites in the perianal region

A

At night, when the body is at rest, gravid (pregnant) adult female worms exit the host, typically a child, through the rectum and lay numerous eggs in the perianal region.

118
Q

cellophane swab:
time of collection

A

early in the morning
before the patient washes or defecates

119
Q

Standard protocol for specimens collected daily for the number of negative tests that should be performed to rule out a pinworm infection is ______.

A

five

120
Q

Each worm lays about how many eggs? and come infective in how many hours?

A

10,000-11,000 eggs
4-6 hours

121
Q

T/F:
Laboratory technicians may need to explain the procedure to patients, their families, and/or other health care professionals.

A

True

122
Q

When instructing others, why is it critical to emphasize the importance of exercising proper hygiene and preventive measures during specimen collection?

A

to avoid spreading infectious eggs into the environment.

123
Q

Explain cellophane swab collection procedure

A
  1. loop tape over end of wooden tongue depressor to expose adhesive surface
  2. touch adhesive surface several times to perianal region
  3. smooth tape on slide
  4. apply drop of toluol or I2 in xylol
124
Q

what solution is used in cellophane swab examination

A

toluol or I2 in xylol

125
Q

Enterobius vermicularis:
worm type according to egg-laying capacity

A

oviparous

126
Q

Enterobius vermicularis:
shape

A

D-shaped

127
Q

Enterobius vermicularis:
size

A

50-60 x 20-30um

128
Q

Enterobius vermicularis:
color

A

colorless

129
Q

Enterobius vermicularis:
shell

A

thin

130
Q

Enterobius vermicularis:
contents

A

single cell (immature egg) or
larva (mature egg)

131
Q

other terms for adhesive tape test

A

transparent tape test
scotch test

132
Q

method of choice to detect Enterobius vermicularis (pinworm) eggs and occasionally adult females

A

adhesive tape test

133
Q

2 types of concentration techniques

A

sedimentation
flotation

134
Q

procedures that allow for detection of parasitic elements

A

concentration (sedimentation and flotation)

135
Q

Why should concentration procedures be carried out?

A

parasitic elements might be missed when examining only a direct wet smear

136
Q

procedure that leads to recovery of all protozoan cysts and oocysts, helminth eggs and larvae present in stool specimen

A

Formalin-ethyl acetate sedimentation concentration

137
Q

the easiest to perform
least subject to technical error allows broad recovery of parasitic elements

A

Formalin-ethyl acetate sedimentation concentration

138
Q

Type of stool specimen for formalin-ethyl acetate sedimentation concentration

A

fresh or fixed

139
Q

T/F:
Formalin-ethyl acetate sedimentation concentration preparation will often contain more debris than that obtained with the flotation and other procedures.

A

True

140
Q

Formalin-ethyl acetate sedimentation concentration is not recommended for?

A
  1. eggs of Fasciola spp.
  2. larvae of Strongyloides stercoralis
141
Q

sediment and surface examinations:
specific gravity

A

sediment:
solution = lower SG than parasite

surface:
solution - higher SG than parasite

142
Q

In surface examination, why should the solution have a higher specific gravity than the parasite?

A

for the parasite to float

143
Q

sedimentation mechanism

A

parasite will go to sediment

144
Q

concentration technique:
easier to perform
applicable to all parasites
routinely performed

A

sedimentation

145
Q

concentration technique:
clean procedure

A

floatation

146
Q

Before: Formalin Ether Concentration Technique
Now: ?

A

FEACT: Formalin Ether-Ethyl Acetate Concentration Technique

147
Q

dissolve lipid contents and other debris

A

ethyl acetate

148
Q

Cite steps for sedimentatation

A
  1. mix sample with fixative
  2. strain suspension and centrifugate
  3. remove supernatant and break sediment
  4. add saline to sediment, seal tube
  5. add ethyl acetate and shake vigorously
  6. wait, remove stopper
  7. centrifuge
  8. contents will separate
  9. detach plug of debris from tube wall, pour off the top 3 layers by invertion
  10. mix sediment with remaining liquid
  11. place a sediment drop on slide, cover with coverslip, place lugol-stained preparation
  12. examine using microscope
149
Q

4 layers in sedimentation

A
  1. ethyl acetate
  2. bulk debris
  3. saline
  4. sediment
150
Q

allows separation of parasitic elements from the coarest organic debris

A

floatation technique

151
Q

type of stool for floatation technique

A

fresh or fixed

152
Q

most widely used floatation solutions

A

zinc sulfate solution
sodium chloride

153
Q

eggs that are not efficiently concentrated with floatation technique

A

heavy eggs:
Fasciola eggs
infertile Ascaris eggs

154
Q

In floatation technique, eggs and cysts tend to lose their typical shape after how long?

A

40-60 minutes

155
Q

4 layers in floatation technique

A
  1. meniscus
  2. eggs/ (oo)cysts
  3. floatant solution
  4. sediment
156
Q

Cite steps for floatation

A
  1. add sample to formalin, mix, leave for at least 30 mins
  2. filter suspension
  3. centrifuge, discard supernatant
  4. resuspend sediment in saline, repeat step 3
  5. resuspend sediment in solution, centrifuge
  6. remove tube from centrifuge, add drops of solution until meniscus forms
  7. harvest upper part of meniscus by placing coverslip, pace it face down on a slide
  8. examine using a microscope
157
Q

In floatation technique, a few drops of this may be added to the slide to enhance morphological details of parasites (e.g. Giardia cysts)

A

Lugol’s solution

158
Q

10 miscellaneous concentration techniques

A
  1. Harada-Mori Culture
  2. Baermann Culture
  3. Knott’s Concentration Technique
  4. Sheather’s Flotation Technique
  5. Brine Flotation Technique
  6. McMaster Concentration Technique
  7. Mini-FLOTAC Technique
  8. Rapid Diagnostic Test for Malaria
  9. Circumoval Precipitin Test (COPT)
  10. Sabin-Feldman Dye Test
159
Q

concentration technique:
- placing stool sample onto filter paper
- adding water
- examining water for filariform larvae detection

A

Harada-Mori Culture

160
Q

concentration technique:
- sealing of funnel
- adding water
- stool wrapped up in cheesecloth
- Filariform larva will go out of cheesecloth
- examine water for larva presence
- uses more stool

A

Baermann Culture

161
Q

concentration technique:
- blood concentration technique used for microfilaria
- add 20 mL of 2% formalin
- centrifuge at 500g for 10 minutes
sediment of conc. blood to slide (thick film)

A

Knott’s Concentration Technique

162
Q

stain used in Knott’s Concentration Technique

A

giemsa

163
Q

concentration technique:
used for coccidians to recover oocyst

A

Sheather’s Flotation Technique

164
Q

concentration technique:
involves mixing the sample with a salt solution to make the eggs and cysts float to the surface

A

Brine Flotation Technique

165
Q

concentration technique:
involves diluting the fecal sample and then using a specialized counting chamber, which has two chambers and grid markings

A

McMaster Concentration Technique

166
Q

concentration technique:
involves mixing the sample with a flotation solution and using a specialized chamber; parasite eggs or oocysts float to the top

A

Mini-FLOTAC Technique

167
Q

concentration technique:
reacts through serological immunoassay

A

Rapid Diagnostic Test for Malaria

168
Q

concentration technique:
confirmatory test used for Schistosoma

A

Circumoval Precipitin Test (COPT)

169
Q

sample for COPT

A

serum

170
Q

serum incubation period for COPT

A

24 hours

171
Q

concentration technique:
for Toxoplasma gondii infection from cat stool

A

Sabin-Feldman Dye Test

172
Q

vulnerable patients to Toxoplasms gondii infection

A

pregnant women

173
Q

mechanism of Toxoplasma gondii infection

A

can cause placental barriers
can cause blindness
can cause cerebral defects in developing fetus

174
Q

sample for Sabin-Feldman Dye Test

A

serum

175
Q

dye used in Sabin-Feldman Dye Test

A

methylene blue dye

176
Q

Sabin-Feldman Dye Test results

A

positive: dye did not bind to trophozoites
negative: dye bound to trophozoites

177
Q

MOT of Toxoplasma gondii infection

A

ingestion of oocyst from cat feces

178
Q

T/F:
Motility of trophozoites is best demonstrated using Lugol’s iodine

A

False;
use NSS

179
Q

T/F:
Improper placement of coverslip often leads to the formation of air bubbles in the fecal smear.

A

True

180
Q

Identify: What reagent acts as a clearing agent in the Kato-Katz preparation?

A

Glycerine-glycerol

181
Q

Identify: Currently, what is considered as the gold standard for the diagnosis of Malaria?

A

Thick and thin blood smears

182
Q

Identify: What is the specimen of choice for the diagnosis of pinworm infections?

A

cellophane swab

Parasitology-Lab (8 decks)

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