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Parasitology > CHAPTER 2 (SPECIMEN COLLECTION AND PROCESSING) > Flashcards

CHAPTER 2 (SPECIMEN COLLECTION AND PROCESSING) Flashcards

Specimen Collection and Processisng

1
Q

the most common procedure performed in the area of parasitology is the examination of

A

stool specimen for ova and
parasites

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2
Q

There are two general components associated with O and P

A

macroscopic and microscopic examination

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3
Q

The microscopic examination consists of
three possible components

A

collection, transport, and fixatives for preservation.

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4
Q

Morphologic forms of protozoa and helminths may be detected from a properly collected and prepared stool specimen. When present, the protozoan forms known as

A

trophozoites and cysts

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5
Q

eggs, larvae, proglottids are collectively known as

A

Helminth stages

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6
Q

The typical stool collection protocol consists
of

A

three specimens, one specimen collected
every other day or a total of three collected in 10 days.

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7
Q

One exception of the typical stool collection protocol is in the diagnosis of

A

amebiasis in which up to six specimens
in 14 days is acceptable

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8
Q

Certain medications and substances may
interfere with the detection of parasites. Stool samples from patients whose therapy includes

A

barium, bismuth, or mineral oil

patients who have taken antibiotics
or antimalarial medications

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9
Q

How many days should pass before collecting stool sample from patients who have treatments that contain barium, bismuth, or mineral oil?

A

5 to 7

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10
Q

Collection of specimens
from patients who have taken antibiotics
or antimalarial medications should be delayed for?

A

2 weeks

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11
Q

Stool specimens should be collected in a

A

clean, watertight container with a tight-fitting lid

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12
Q

The acceptable amount of stool required for parasite study is

A

2 to 5 g

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13
Q

should not be allowed to contaminate
the stool specimen because it has been
known to destroy some parasites

A

Urine

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14
Q

Stool should not be retrieved from ________ water because free-living protozoa and nematodes may be confused with human parasites

A

toilet bowl

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15
Q

water may destroy select parasites, such as

A

schistosome eggs and amebic trophozoites.

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16
Q

Toilet paper in the stool specimen may?

A

mask parasites or make examination of the sample difficult.

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17
Q

The specimen container should be labeled
with

A

the patient’s name and identification number,
the physician’s name, and the date and time of sample collection.

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18
Q

The specimen should be placed
into _____________ for transport to the
laboratory.

A

a zip lock plastic bag

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19
Q

When handling all specimens, __________ should be worn at all times.

A

gloves and a
protective coat

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20
Q

__________ should also be used in laboratories,
when present.

A

Biohazard hoods

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21
Q

Universal precautions, as
outlined by the _________ for handling blood and body fluids, should be exhibited and enforced at all times.

A

Occupational Safety and Health
Administration (OSHA)

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22
Q

Another important consideration in testing
fecal specimens for parasites is the

A

time frame from sample collection to receipt and examination in the laboratory

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23
Q

______________ is sensitive to environmental changes and, on release from the body, disintegrates rapidly thus a fresh sample is required

A

trophozoite stage

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24
Q

fresh specimen of protozoan trophozoites is required in order to?

A

demonstrate the motility

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25
Q

it is recommended
that liquid specimens be examined
within _______ because trophozoites are usually found in this type of stool

A

30 mins of passage

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26
Q

In keeping with stool consistency, semiformed specimens may yield a mixture of protozoan cysts and trophozoites and should be evaluated within?

A

1 hour of passage

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27
Q

Formed stool specimens are not likely
to contain trophozoites; therefore, they can be held for

A

24 hours following collection

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28
Q

The specimen can be preserved by placing it directly into a ______ at the time it is collected or on ______

A

fixative
receipt in the laboratory

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29
Q

How many stool samples should be collected when following the typical O&P collection protocol?
A. 1
B. 2
C. 3
D. 4

A
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30
Q

are substances that preserve the morphology of protozoa and prevent further development of certain helminth eggs and larvae

A

Fixatives

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31
Q

whatever fixative is used, the recommended ratio is

A

three parts fixative to one part stool
3:1

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32
Q

The specimen must be
fixed in the preservative for at least ______ before processing

A

30 minutes

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33
Q

has been used for many years as an all-purpose fixative for the recovery of protozoa and helminths.

A

Formalin

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34
Q

Two concentrations
of formalin are commonly used; a

A

5% and 10%

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35
Q

5% concentration ideally preserves

A

protozoan cysts

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36
Q

10% concentration preserves

A

helminth eggs and larvae

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37
Q

Formalin may be routinely used for

A

direct examinations
and concentration procedures

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38
Q

Formalin is not used for?

A

permanent smears

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39
Q

There are three primary advantages for the use of formalin

A

(1)it is easy to prepare

(2) it preserves specimens for up to several years

(3) it has a long shelf life

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40
Q

disadvantages of formalin
is that

A

it does not preserve parasite morphology
adequately for permanent smears.

trophozoites usually cannot be recovered and morphologic details of cysts and eggs may fade with time

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41
Q

requirements to use formalin under OSHA regulations

A

Monitoring of vapors, use of protective clothing, and a comprehensive, written chemical hygiene plan (CHP)

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42
Q

Concentration, Permanent stain, and Antigen Tests of 10% formalin

A

(+) (-) (+)

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43
Q

Concentration, Permanent stain, and Antigen Tests of SAF

A

(+) (+) iron hematoxylin (+)

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44
Q

Concentration, Permanent stain, and Antigen Tests of PVA

A

(±) (+) trichrome or iron hematoxylin (-)

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45
Q

Concentration, Permanent stain, and Antigen Tests of Modified PVA (zinc)

A

(±) (+) trichrome or iron hematoxylin (±)

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46
Q

Concentration, Permanent stain, and Antigen Tests of Modified PVA (zinc) Single-vial system

A

(+) (+) trichrome or iron hematoxylin (±)

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47
Q

comprised of a plastic powder that acts as an
adhesive for the stool specimen when preparing slides for staining

A

Polyvinyl Alcohol

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48
Q

PVA is most often combined
with

A

Schaudinn solution, which usually contains
zinc sulfate, copper sulfate, or mercuric chloride as a base

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49
Q

What can PVA detect

A

Trophozoites and cysts of the protozoa, as well as most helminth eggs

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50
Q

The greatest advantage of PVA is

A

preparation of a permanent stained smear

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51
Q

The biggest disadvantage of the use of PVA
is that

A

Schaudinn solution contains mercuric chloride.

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52
Q

This preservative can be used for performing concentration techniques and permanent stained smears. Some laboratories have adopted this fixative because it only requires a single vial and it is mercury-free.

A

Sodium Acetate Formalin

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53
Q

Advantages of SAF

A

SAF is
easy to prepare

long shelf life

used for preparing smears for staining with the modified acid-fast stain to detect coccidian oocysts.

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54
Q

SAF also has disadvantages because

A

adhesive is not good

protozoa morphology not as clear

limited choice of permanent stains

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55
Q

what is added to aid SAF for it to adhere

A

albumin

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56
Q

Other alternatives to mercury-based PVA are the use of substitute compounds containing copper sulfate or zinc sulfate.

A

Modified Polyvinyl Alcohol

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57
Q

advantage and disadvantage of Modified Polyvinyl Alcohol

A

can be used for concentration methods
and permanent stained smears

substitute products do not provide the same
quality of preservation for adequate protozoan morphology on a permanent stained slide as the mercury-based fixatives

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58
Q

Several manufacturers have developed alternative nontoxic fixatives. These single-vial fixatives are free of formalin and mercury and can be used for concentration techniques and permanent stained smears.

A

Alternative Single-Vial Systems.

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59
Q

disadvantage of Alternative Single-Vial Systems.

A

do not provide the same quality of preservation as mercury-based fixatives and organism identification will be more difficult from permanent stained slides.

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60
Q

advantages of single vial systems

A

can also be used for performing fecal immunoassays

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61
Q

What is the purpose of fixatives for the collection of
stool samples?
A. Enhance the motility of protozoa.
B. Stain the cytoplasmic inclusions of protozoa.
C. Preserve the morphology of protozoa and prevent
further development of helminths.
D. All of the above.

A
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62
Q

Once a stool specimen has been received in the laboratory, the analytic phase of laboratory testing, also referred to __________ begins

A

processing

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63
Q

Stool specimens submitted for parasitic study should first be examined ________ to determine the consistency and color of the sample. The specimen should be screened and examined for the presence of gross abnormalities.

A

macroscopically

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64
Q

Macroscopic examination stool samples should be?

A

fresh, unpreserved stool specimen

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65
Q

In such situations where macroscopic examination is not possible what is recommended at the time of specimen collection.

A

a notation of the gross appearance, either on the actual specimen container or on the requisition form

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66
Q

What type of parasites are potentially present in

Liquid Stools?
Fully Formed Stools?
Both?

A

Liquid Stools - Protozoan trophozoites
Fully Formed Stools - Protozoan cysts
Both - Helminth eggs and larvae

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67
Q

What physical characteristics of a stool indicates the patient’s condition such as whether a patient has recently had a special procedure or if the patient is on antibiotic therapy?

A

Color of the stool

68
Q

Range of colors of stool

A

Black to Green to Clay
Brown is normal
Unusual colors = particular medication

69
Q

Gross abnormalities possibly found in stool
include?

A

adult worms, proglottids, pus, and
mucus

70
Q

First step of macroscopic examination of stools

A

surface of the stool should be examined for parasites, such as pinworms, tapeworm proglottids, and adult worms

71
Q

Second step of macroscopic examination of stools

A

sample should then be broken up and examined once more for macroscopic parasites, especially adult helminths.

(a wooden applicator stick works nicely for this task)

72
Q

What to do if adult worms are found in the sample

A

sample should be carefully washed through a wire screen

73
Q

Other macroscopic abnormalities in the specimen may have parasitic indications. For example,

blood and/or mucus in loose or liquid stool may suggest the presence of

A

amebic ulcerations in the
large intestine

74
Q

Bright red blood on the surface
of a formed stool is usually associated with

A

irritation and bleeding

75
Q

Which of the following characteristics is observed during the macroscopic examination of stool specimens?

A. Consistency
B. Color
C. Adult worms
D. All of the above

A
76
Q

Macroscopic Examination of
Stool Specimens: Possible
Descriptive Terms

Consistency Terms (IN ORDER)

A

Hard
Soft
Mushy
Loose
Diarrheic
Watery, liquid
Formed
Semiformed

77
Q

Macroscopic Examination of
Stool Specimens: Possible
Descriptive Terms

Possible Colors (IN ORDER)

A

Dark brown
Black
Brown
Pale brown
Clay
Yellow
Red-brown
Green, other

78
Q

Macroscopic Examination of
Stool Specimens: Possible
Descriptive Terms

Gross Appearance Terms (IN ORDER)

A

Conspicuously fibrous
Fiber scanty to moderate
Colloidal (homogeneous)
Scanty mucus
Much mucus
Mucus with scanty blood
Other (e.g., blood, barium)

79
Q

To detect the presence of parasites in a stool specimen, ______are performed.

A

microscopic examinations

80
Q

The microscopic examination of stool for ova and parasites which uses fresh samples involves three distinct procedures which are

A

direct wet preparations (preps)

concentrated technique
resulting in concentrated wet preparations

permanently stained smear

81
Q

If the specimen is received in fixative what step in the microscopic examination can be eliminated?

A

direct wet preparation

82
Q

Which of these procedures is involved in the microscopic
examination of stool specimens for parasites?

A. Performing a concentration technique
B. Determining specimen consistency
C. Examining sample for gross abnormalities
D. Analyzing sample for color

A
83
Q

After the three steps in the microscopic examination, what is done next?

A

concentrate and permanent stain techniques are performed

84
Q

The most important piece of equipment
in the parasitology laboratory is the

A

microscope

85
Q

Because size is an important diagnostic
feature in parasitology, it is necessary for
the microscope to contain a specially designed ocular piece equipped with a measuring scale known as an

A

ocular micrometer

86
Q

The diagnostic stages of parasites detected
microscopically are measured in units known
as

A

microns

μ or μm

0.001 [10−3] millimeter

10^−6 meter

87
Q

An important step in order to ensure accurate measurements of the microscope and what tool it uses

A

calibration

stage micrometer

88
Q

defined as a slide made by mixing a small portion of unfixed stool with saline or
iodine and subsequent examination of the resultant mixture under the microscope

A

direct wet preparation

/direct wet mount

89
Q

Direct wet preparations is used to?

A

detect the presence of motile protozoan trophozoites.

90
Q

Other parasite
stages that might be observed in a direct wet
preparation include

A

protozoan cysts, oocysts, helminth eggs, and larvae.

91
Q

First step in direct saline wet preparation

A

placing a drop of 0.85% saline on a glass slide
and mixing with a small portion of unfixed stool using a wooden applicator stick or another mixing tool.

92
Q

Second step in direct saline wet preparation

A

A 22-mm square cover slip is placed on the slide and the preparation is examined microscopically in a systematic fashion

93
Q

Third step in direct saline wet preparation

A

The entire cover glass should be scanned using the low power (10×) objective on the microscope, and the power should only be increased when a suspicious object requires further investigation.

94
Q

Performing this procedure allows for the ability to observe greater detail using the 100× objective.

A

oil immersion

95
Q

is not usually recommended on
wet preparations unless the cover slip is sealed to the slide.

A

oil immersion

96
Q

How to seal cover slip to the slide

A

A temporary seal can be prepared
using a hot paraffin-petroleum jelly (Vaseline)
mixture around the edges of the cover slip

97
Q

may be made
to enhance the detail of protozoan cysts

A

direct iodine wet preparation

98
Q

what iodine formulas are used for a direct iodine wet preparation

A

Lugol’s or D’Antoni’s formula)

99
Q

Which are killed when using a direct iodine wet preparation?

A

trophozoites

100
Q

What are the proper adjustments for the microscope in order to view even transparent protozoa?

A

light should be reduced using the
iris diaphragm - (provide contrast between the cellular elements)

Lowering the condenser - (allow transparent
structures to be seen)

101
Q

The direct wet preparation can be eliminated from the O&P examination if the specimen is received in a fixative.
A. True
B. False

A
102
Q

provide the ability to detect small numbers of
parasites that might not be detected using direct wet preparations

A

Concentration techniques

(or methods? idk same same maybe)

103
Q

The purpose of concentration
is to

A

aggregate parasites

remove as much debris

104
Q

Concentration techniques examination allows
the laboratory technician to detect

A

protozoan cysts, oocysts, helminth eggs, and larvae

105
Q

do not usually survive concentration techniques

A

Protozoan trophozoites

106
Q

There are two types of concentration methods available

A

sedimentation and flotation

107
Q

Most experts recommend what concentration technique be used, because it is more efficient and easier to perform accurately.

A

sedimentation

108
Q

The most widely used sedimentation
technique is the

A

formalin–ethyl acetate sedimentation
procedure

109
Q

The advantage of formalin–ethyl acetate sedimentation procedure is that it

A

good recovery of most parasites
and is easy to perform

110
Q

The disadvantage of formalin–ethyl acetate sedimentation procedure is that it

A

preparation contains
more fecal debris than a flotation technique

111
Q

is also based on differences
in specific gravity between the sample
debris, which in this case is heavy and sinks to
the bottom of the test tube, and potential parasites, which are lighter and float toward the top of the tube

A

Zinc Sulfate Flotation Technique

112
Q

Zinc Sulfate Flotation Technique uses? (include specific gravity)

A

zinc sulfate with a specific gravity of 1.18 to 1.20

113
Q

The advantage of Zinc Sulfate Flotation Technique

A

more fecal debris is removed
and it yields a cleaner preparation

114
Q

The disadvantage
of Zinc Sulfate Flotation Technique

A

some helminth eggs are very dense and will not float

115
Q

It is recommended
that if laboratories perform zinc sulfate flotation technique that they

A

examine saline and iodine preps made from the sediment microscopically, as well as the surface film (direct wet preparation samples)

116
Q

Which of the following parasitic stages is not usually
detected after using a concentration technique?
(Objective 2-9)
A. Protozoan cysts
B. Protozoan trophozoites
C. Helminth eggs
D. Helminth larvae

A

B. Protozoan trophozoites

117
Q

defined as a microscope slide that
contains a fixed sample that has been allowed to dry and subsequently stained

A

permanent stained smear

118
Q

The final procedure in the O&P examination is the preparation and examination of a

A

permanent stained smear

119
Q

Permanent staining samples is a critical portion of the O&P examination because it is designed to

A

confirm the presence of protozoa cysts and/or trophozoites

120
Q

Example of protozoa that only possess a trophozoite stage and will not be detected in the concentrated wet mount preparation that is why permanent stains are important

A

Dientamoeba fragilis

121
Q

The permanent stained smear is not the
method of choice for the identification of _______ because these parasites often
stain too dark or appear distorted

A

helminth eggs or larvae

122
Q

helminth eggs or larvae are best detected and identifies using what technique?

A

concentration technique

123
Q

The sample of choice for such stains is a
thinly prepared slide of see-through thickness
made from a

A

PVA-preserved sample

124
Q

Specimens fixed with SAF may also be used in permanent stain techniques, but the choice of stain is limited to

A

iron hematoxylin

125
Q

Slides can also be prepared from a
fresh stool specimen but?

A

Should not be dried and fixed immediately

126
Q

Example of a fixative used to samples with fresh stool in permanent stain techniques

A

Shaudinn fixative

127
Q

How many fields are reviewed before the slide can be considered negative in permanent stain techniques?

A

300 fields

128
Q

Two common stains used for routine O&P
testing include

A

trichrome (Wheatley modification)

iron hematoxylin

129
Q

are not part of the routine O&P examination
and must be specifically requested

A

Specialized stains

130
Q

These
specialized stains include what stains?

A

modified acid-fast
and modified trichrome stains.

131
Q

What structure or material appear LIGHT PINK OR BLUE-GREEN on Trichrome Stain

A

Cytoplasm of Entamoeba histolytica trophozoites and cysts

132
Q

What structure or material appear PURPLE TINT on Trichrome Stain

A

Cytoplasm of Entamoeba coli cysts

133
Q

What structure or material appear BRIGHT RED TO RED-PURPLE on Trichrome Stain

A

Nuclear karyosomes

134
Q

What structure or material appear LIGHT GREEN on Trichrome Stain

A

Degenerated parasites

135
Q

What structure or material appear GREEN on Trichrome Stain

A

Background

136
Q

The most widely used
permanent stain is the

A

Wheatley trichrome stain

137
Q

may be used instead of the trichrome technique. Historically, this procedure was considered to be time-consuming. However, a shorter technique using this stain is now available

A

iron hematoxylin
stain

138
Q

The iron hematoxylin stain
reveals excellent morphology of the

A

intestinal protozoa

139
Q

Appearance of Select
Protozoan Structures and
Background Material on
IRON HEMATOXYLIN Stain

BLUE TO PURPLE

A

Protozoa cytoplasm

140
Q

Appearance of Select
Protozoan Structures and
Background Material on
IRON HEMATOXYLIN Stain

DARK BLUE TO DARK PURPLE

A

Protozoa Nuclear Material

141
Q

Appearance of Select
Protozoan Structures and
Background Material on
IRON HEMATOXYLIN Stain

LIGHT BLUE (sometimes with pink tint)

A

Debris and Background Material

142
Q

Appearance of Select
Protozoan Structures, Yeast,
and Background Material
on MODIFIED ACID-FAST STAIN

PINK TO RED

A

Oocysts of Cryptosporidium and Isospora

143
Q

Appearance of Select
Protozoan Structures, Yeast,
and Background Material
on MODIFIED ACID-FAST STAIN

VARIABLE; CLEAR TO PINK RED

A

Oocysts of Cyclospora

144
Q

Appearance of Select
Protozoan Structures, Yeast,
and Background Material
on MODIFIED ACID-FAST STAIN

BLUE

A

Yeast

145
Q

Appearance of Select
Protozoan Structures, Yeast,
and Background Material
on MODIFIED ACID-FAST STAIN

BLUE OR LIGHT RED

A

Background Material

146
Q

Disadvantage of specialized stains

A

do not detect oocysts of the coccidian parasites or spores of microsporidia

147
Q

has become an important permanent
stain procedure for the detection of the oocysts of Cryptosporidium, as well as those of Isospora and Cyclospora

A

modified acid-fast stain

148
Q

this allows for the detection of acid-fast
parasites in addition to the other protozoa normally recovered using the iron hematoxylin stain

A

a carbol fuchsin step incorporated to a modified iron hematoxylin stain

149
Q

a carbol fuchsin step incorporated to a modified iron hematoxylin stain uses what samples

A

SAF-preserved fecal samples

150
Q

Although the spores of microsporidia will also stain with the acid-fast technique, their small size (1 to 2 μm) makes it difficult to identify them without the use of special stains. What modified stain are available to demonstrate these parasites

A

Modified Trichrome Stains

151
Q

Appearance of Microsporidia on MODIFIED TRICHROME STAIN

PINK TO RED WITH CLEAR INTERIOR

A

Spores of microsporidia

152
Q

Appearance of Microsporidia on MODIFIED TRICHROME STAIN

RED HORIZONTAL OR DIAGONAL BAR

A

Polar Tubule

153
Q

Appearance of Microsporidia on MODIFIED TRICHROME STAIN

PINK TO RED

A

Bacteria, Yeast, Debris

154
Q

Appearance of Microsporidia on MODIFIED TRICHROME STAIN

GREEN

A

Background

155
Q

These methods can be obtained as kits that contain monoclonal antibody. This commercial antibody is used to detect
antigens in patient specimens with the main purpose of ease and speed.

A

rapid methods, or stool-screening methods.

156
Q

Current available assays of stool screening methods include

A

enzyme immunoassay (EIA), direct fluorescent antibody (DFA), and membrane flow cartridge techniques.

157
Q

The antigen detection methods on stool-screening methods are commercially available for specific intestinal protozoa, including

A

Entamoeba histolytica, Giardia intestinalis, and Cryptosporidium spp.

158
Q

What is one advantage of the stool screening method?

A. It is highly sensitive and specific.
B. It can detect all parasites.
C. It can be performed on fresh or preserved
specimens.
D. It is labor-intensive.

A

A. It is highly sensitive and specific.

159
Q

The permanent stained smear is critical for detection
of helminth eggs and larvae.
A. True
B. False

A
160
Q

When reporting a positive specimen, the
report should state

A

scientific name
stage of development of the parasite

also helpful:
presence of certain cells (White blood cells should be reported semiquantitatively)

161
Q

The results of the O&P procedure should
include a comment indicating that

A

this procedure does not detect Cryptosporidium spp., Cyclospora
cayetanensis, and microsporidia; it will recover the oocysts of Isospora belli

162
Q

The results of fecal immunoassays should indicate the

A

specific parasite(s) that is (are) tested for in the assay

163
Q

In general, for most parasites, quantitation is
highly required

true or false

A

false - not indicated

164
Q

Situations in which quantitation of parasite is important are as follows

A

Blastocystis hominis
helminth eggs
Trichuris trichiura
Clonorchis sinensis
Schistosoma spp.
Plasmodium spp.
Babesia spp.

165
Q

These are crystals which are reported and quantitated when found

A

Charcot-Leyden crystals

166
Q

Which one of these parasites should be quantitated
in the parasitology report?
A. Giardia intestinalis
B. Entamoeba coli
C. Trichomonas vaginalis
D. Blastocystis hominis

A

D. Blastocystis hominis

Parasitology (6 decks)

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