Semen Analysis - Brunzel Prt. 2

Question 1

Because sperm concentration in normal seminal fluid can vary significantly, ______samples should be analyzed to evaluate male fertility.

Answer 1

two or more samples

Question 2

Specimen collections should take place within a _____period and at least_____ apart.

Sexual abstinence for at least______, but not more than _____, should precede the collection.

Answer 2

3-month ; 7 days

2 days (48 hours), but not exceeding 7 days

Question 3

The patient collects the specimen through ______, and the entire ejaculate is collected in a________ container.

Although some plastic containers are toxic to spermatozoa, others are not.

Answer 3

masturbation

clean, wide-mouth sterile plastic or glass container

Question 4

The collection container should be kept at _______ or ________before the collection to avoid the possibility of cold shock to the sperm.

The container can be warmed easily by holding it next to the ________for several minutes before the collection.

This technique can also be used to control the temperature of specimens being transported in cold climates.

Answer 4

room temperature or warmed (to approximately body temperature)

patient’s body or under the arm

Question 5

Specimen containers and request forms must be labeled with the (3).

Answer 5

patient’s name

the period of sexual abstinence

and the date and time of specimen collection

Question 6

The time of actual specimen collection is crucial in evaluating_____ and _____

Answer 6

liquefaction and sperm motility

Question 7

During specimen collection, ________ or ______ should not be used because they have spermicidal properties.

For patients unable to collect a specimen through masturbation, special nonspermicidal (e.g.,______) condoms can be provided for specimen collection.

Answer 7

lubricants and ordinary condoms

Silastic condoms

Question 8

The collection of seminal fluid requires sensitivity and professionalism.

_____ and _____should be provided to the patient, as well as a comfortable and private room near the laboratory.

If the specimen is to be collected elsewhere and delivered to the laboratory, clearly written instructions regarding specimen transport conditions must be provided

Answer 8

Written and verbal instructions

Question 9

Specimens must be received in the laboratory ______ after the collection, and they must be protected from extreme temperatures, that is, maintained at______ to ____.’

If these criteria are not met, the specimen will not be satisfactory for sperm function tests and an abnormally low sperm motility can result.

Answer 9

within 1 hour

20°C to 40°C

Question 10

When a portion of the initial ejaculate is not collected (3)

Answer 10

the sperm concentration will be falsely decreased

owing to a reduction in prostate secretions, the pH is falsely increased

the coagulum will fail to liquefy

Question 11

When the last portion of an ejaculate is missing (primarily seminal vesicle fluid)

(4)

Answer 11

the semen volume will be decreased

the sperm concentration falsely increased

the pH falsely decreased

a coagulum will not form

Question 12

As with all body fluids, seminal fluid represents a potential biohazard and must be handled accordingly.

Because seminal fluid can contain infectious agents such as (3), and others, all personnel must adhere to standard precautions when handling these specimens.

Answer 12

hepatitis virus

human immunodeficiency virus

herpes virus

Question 13

Semen Characteristics Associated With Fertility

Physical Examination (3)

Answer 13

Appearance
Volume
Viscosity/liquefaction

Question 14

Semen Characteristics Associated With Fertility

Microscopic Examination (5)

Answer 14

Motility
Concentration
Morphology
Vitality
Leukocytes

Question 15

Semen Characteristics Associated With Fertility

Chemical Examination (5)

Answer 15

pH
Acid phosphatase (total)
Citric acid (total)
Fructose (total)
Zinc (total)

Question 16

______ is gray-white and opalescent in appearance.

A______ may indicate the presence of blood,

whereas_____ coloration has been associated with certain drugs.

Answer 16

Normal semen

brown or red hue

yellow

Question 17

If large numbers of leukocytes are present, the semen appears more_______

When the specimen appears almost_____, the sperm concentration is usually low.

Answer 17

turbid with less translucence

clear

Question 18

Semen has a distinctive odor that is sometimes described as_____.

Although infections in the male reproductive tract can modify this odor, a change is rarely noted or reported.

Answer 18

musty

Question 19

Semen is a homogeneous, viscous fluid that immediately coagulates after ejaculation to form a_____.

Within_____, it liquefies (becomes watery).

Although liquefaction can take longer, any delay beyond_____ is considered abnormal and must be noted.

Answer 19

coagulum

30 minutes

60 minutes

Question 20

Because______ is necessary to perform analysis, semen specimens that do not liquefy completely can be chemically treated.

After normal liquefaction, undissolved gel-like granules or particles can be present in the specimen, with a small amount considered normal.

Answer 20

complete liquefaction

Question 21

The ______ and ______analyses of seminal fluid should take place immediately after liquefaction or within 1 hour after collection (for specimens collected away from the laboratory).

Answer 21

physical and microscopic

Question 22

Specimen volume is measured to one decimal place (_____) using a______ (5.0 mL or 10.0 mL).

If a semen culture for bacteria is requested, the volume measurement should be performed first using sterile technique.

Answer 22

0.1 mL

sterile serologic pipette

Question 23

Normally, a complete ejaculate collection recovers_______ of seminal fluid.

Volumes less than and greater than this range are considered abnormal and have been associated with infertility.

Answer 23

2 to 5 mL

Question 24

After complete liquefaction, the_____ of the semen is evaluated using a______ and observing the droplets that form when the fluid is allowed to fall by gravity.

Answer 24

viscosity

Pasteur pipette

Question 25

Normal viscosity of semen

Answer 25

Watery

Question 26

Abnormal viscosity or fluid thickness is indicated by the formation of a_________ A high content of____ can increase the viscosity.

Answer 26

string or thread greater than 2cm in length mucus

Question 27

Other conditions associated with increased viscosity include the production of________ and______ (i.e., decreased concentration and motility of sperm).

Answer 27

antisperm antibodies oligoasthenospermia

Question 28

Lower Reference Limit of VOLUME

Answer 28

1.5 mL (1.4-1.7)

Question 29

Some laboratories use a single stain for the evaluation of several parameters, such as _______ stain for sperm vitality, morphology, and the identification of other cells, whereas others use different stains that specifically enhance each parameter to aid in the identification and evaluation of sperm and other cells.

Answer 29

eosin-nigrosin

Question 30

______ is one of the most important characteristics of sperm because_____, even in high concentrations, are unable to reach and fertilize an ovum.

Answer 30

Motility immotile sperm

Question 30

Some laboratories use a single stain for the evaluation of several parameters, such as _______ stain for sperm vitality, morphology, and the identification of other cells, whereas others use different stains that specifically enhance each parameter to aid in the identification and evaluation of sperm and other cells.

Answer 30

eosin-nigrosin

Question 31

Without an automated system, sperm motility is evaluated subjectively and semiquantitatively using_______ microscopy (______can also be used with appropriate condenser adjustments). After complete liquefaction, the semen sample is mixed well to ensure homogeneity.

Answer 31

phase-contrast; brightfield microscopy

Question 32

Motility A consistent volume of each specimen is evaluated by pipet-ting a fixed volume (e.g., 10 or 20 L) of semen onto a microscope slide using a calibrated_______. The sample is covered with a coverslip of predetermined size (e.g., 18 x 18 mm), and the slide is allowed to settle for about _____before evaluation. To enhance the accuracy and precision of results, wet mounts of each sample should be prepared and evaluated in duplicate.

Answer 32

positive-displacement pipette 1 minute

Question 33

Because sperm motility is affected adversely by_____ Some laboratories perform the analysis at_____

Answer 33

temperature room temperature (i.e., 22 ÷ 2°C).

Question 34

Motility The____ and _____ progression of each sperm are evaluated. In normal semen evaluated within 60 minutes of collection,_____% of the sperm will show ***moderate to strong linear or forward progression.*** The practice in some laboratories of reassessing sperm motility at additional time intervals serves no purpose and has no clinical signifi-cance. Physiologically or in vivo, sperm leave the seminal fluid within minutes after ejaculation and enter the cervical mucus. Therefore motility on a microscope slide at later time intervals is irrelevant.

Answer 34

speed and forward progression 50% or more

Question 35

For fertility purposes, the actual_____ of sperm is not as important as other characteristics. This fact is supported by studies of fertile men despite____

Answer 35

number low sperm counts (less than 1 million per mL)

Question 36

The concentration of sperm in an ejaculate is considered normal when ___________ of sperm are present; values less than or greater than this range are considered abnormal and are associated with infertility.

Answer 36

20 million to 250 million per mL

Question 37

The variation in the sperm concentration within a single individual can be significant and depends partially on the (3). For these reasons, ______specimens should be evaluated to reliably assess the quantity and quality of an individual's sperm.

Answer 37

period of sexual abstinence viral infection stress multiple semen

Question 38

Manually, the concentration of sperm is determined by using a_____ after preparing an appropriate dilution of the semen. Frequently, a_____ dilution is prepared. If during initial microscopic examination the sperm concentration is noted to be exceptionally high or low, a new dilution can be prepared and mounted. All dilutions should be made using a_______ pipette to deliver the semen quantitatively to a premeasured amount of diluent

Answer 38

hemacytometer 1:20 calibrated positive-displacement pipette

Question 39

Note that a hematology_____ pipette is not accurate for use with seminal fluid because of its viscosity and should not be used. After the hemacytometer is filled with the well-mixed dilution of semen, it is placed in a humidifying chamber and allowed to settle for _______ before counting. The type of hemacytometer, the specimen dilution used, and the areas counted determine the conversion factor necessary to obtain the concentration of sperm in millions per mL

Answer 39

white blood cell (WBC) pipette 3 to 5 minutes

Question 40

In contrast to sperm concentration (____unit), the_____ is the ***total number of sperm present in the entire ejaculate.*** This value, often requested by clinicians, is calculated by multiplying the_______ by the______

Answer 40

sperm per milliliter sperm count sperm concentration (sperm/ mL) by the total volume of the ejaculate.

Question 41

Sperm count =

Answer 41

Sperm concentration (sperm/mL) × Volume of ejaculate (mL)

Question 42

After a vasectomy, the sperm count in semen ideally should be_______-within______ after the procedure. However, studies have shown that nonmotile sperm can be present for as long as ______post vasectomy regardless of the number of ejaculations.

Answer 42

zero-no sperm present (azoospermia); 12 weeks 21 months

Question 43

Post vasectomy In clinical practice, most men (~66%) demonstrate_______ within ______, regardless of the number of ejaculations. Note that the most important feature is not the number of sperm present post vasectomy but the status of their_____.

Answer 43

azo-ospermia within 12 weeks motility

Question 44

The presence of even a single_______ is evidence of an unsuccessful vasectomy (i.e., recanalization of the vas deferens has occurred), whereas low numbers of _____can persist for months in some men (~33%).

Answer 44

"motile" spermatozoon "immotile" sperm

Question 45

-measurement of the sperm head length, width, circumference, and area-enables the generation of objective data. To be considered normal, sperm must meet strict criteria regarding their size and shape, which can be determined by computerized systems or manually using a microscope with a calibrated ocular micrometer.

Answer 45

Sperm morphometry

Question 46

Human sperm have three distinct areas:

Answer 46

head, midpiece, and tail.

Question 47

When viewed from the side, sperm appear to be_______ When viewed from the top, normal human sperm have oval heads that are______ in width and _____in length. Only sperm lying flat should be evaluated and their head length-to-width ratio should be______

Answer 47

arrowhead-shaped 2.5 to 3.5 um width 4.0 to 5.0 um length 1.5 to 1.75.

Question 48

The midpiece, located immediately behind the head, is_____ long and is thicker than the tail but not greater than____ in width. The tail should be slender, uncoiled, and at least____ long.

Answer 48

6 to 7.5 um; 1 um 45 um

Question 49

If a "complete" morphology evaluation is performed, then each spermatozoon is classified using five cate-gories:

Answer 49

normal head defects midpiece defects tail defects cytoplasmic droplet present

Question 50

_______ are usually located in the midpiece region and are considered abnormal if this region is greater than one-third the area of a normal sperm head. The head can contain vacuoles, but they are not considered abnormal unless they occupy more than _____of the head. Note that a single spermatozoon can have multiple defects, and each defect is documented.

Answer 50

Cytoplasmic droplets 20%

Question 51

Staining enhances the visualization of sperm morphology and enables the identification and differentiation of (3) (i.e., spermatids, spermatocytes, and spermatogonia). (3) stains are frequently used.

Answer 51

WBCs epithelial cells of the urethra and immature spermatogenic cells Giemsa Wright's Papanicolaou

Question 52

In some laboratories, a normal sperm morphology of ______ is considered "normal" However, when strict evaluation criteria are used for fertility purposes as in studies of fertile and subfertile individuals, the number of sperm with normal morphology is significantly lower. In these studies, normal sperm morphology of less than ____is a strong predictor of infertility, whereas fertility is associated with normal sperm morphology values of________

Answer 52

50% or greater 5% 12% to 15% or greater.

Question 53

Other variables, particularly (3) provide the greatest predictive value in assessing male fertility.

Answer 53

sperm concentration progressive motility sperm morphology

Question 54

_____ staining of a fresh semen smear enables ***rapid differentiation of live and dead sperm.*** Because dead sperm have damaged plasma membranes, these cells take up stain; living sperm do not

Answer 54

Vital staining

Question 55

_______ or ______ stain (a modification of_____ technique) combination is frequently used to determine sperm vitality. Using brightfield or phase-contrast microscopy and X1000 (or X400) magnification, 100 sperm on a stained smear are evaluated.

Answer 55

Eosin alone or an eosin-nigrosin Blom’s

Question 56

The percentage of_____ sperm cells should not exceed the percentage of_____ sperm. In other words, if 65% of the sperm in a semen specimen are dead, the motility cannot exceed 35%. Hence the vitality evaluation provides a convenient quality or cross-check of the motility evaluation. In fresh normal semen, 50% or more of the sperm are alive.

Answer 56

dead; immotile

Question 57

The spermatogenic cells can be difficult to differentiate from_____ because of size and nuclear pattern similarities. A______ can aid in this evaluation because neutrophils are peroxidase-positive cells, whereas lymphocytes and spermatogenic cells are peroxidase-negative cells. However, owing to the carcinogenicity of the chemicals used in many peroxidase stains and the special handling required,_____ stain may be preferred.

Answer 57

WBCs peroxidase stain Wright's

Question 58

The presence of greater than __________of ejaculate indicates an inflammatory process, most often involving the male accessory glands (e.g., seminal vesicle, prostate). However, a normal WBC count does not rule out infection.

Answer 58

1 million WBCs per milliliter

Question 59

_______, the sticking together of motile sperm, is evident by microscopic examination of a wet preparation. Although some clumping of immotile sperm may occur in normal semen specimens, the observation of a distinct (3) orientation of sperm is associated with the presence of sperm-agglutinat-ing antibodies.

Answer 59

Agglutination head-to-head, head-to-tail, or tail-to-tail

Question 60

(2) antibodies bound to sperm have been identified and correlated with reduced fertility. This is known as _______where the man or the woman produce antisperm antibodies and the individual producing them can be identified. When the man is the source, the antibodies are present on the surface of the sperm______; when the woman is producing them, the sperm are coated with antibodies_____.

Answer 60

Immunoglobulin G and immunoglobulin A antibodies immunologic infertility before intercourse after they enter the cervical mucus

Question 61

Macroscopic and microscopic tests are available to detect and determine the immunoglobulin class of sperm antibodies ([Ig]G, IgA) Both tests produce comparable results, but the _______is rapid (~3 minutes) and easy to perform, whereas the ______t is time-consuming (~45 minutes), technically more complicated, and more expensive. The cutoff values for these tests vary among laboratories.

Answer 61

mixed agglutination reaction (MAR) test immunobead tes

Question 62

The WHO defines agglutination as clinically significant (abnormal) when antisperm antibodies coat ______ or more of the spermatozoa, whereas other institutions use lower cutoffs (e.g., 20%, 10%).

Answer 62

50%

Question 63

The pH of fresh normal semen is alkaline and ranges from_____ to_____. Fresh specimens with a pH less than 7.2 can be obtained from individuals with abnormalities of the (3). In contrast, fresh specimens exceeding pH 7.8 suggest an_____ in the male reproductive tract.

Answer 63

7.2 to 7.8 epididymis, the vas deferens, or the seminal vesicles infection

Question 64

Specimens not tested within 1 hour of collection can show changes in the pH for several reasons. An ***increase in pH*** can occur because of loss of_______; conversely, a ***decrease in pH*** can occur because of the accumulation of_____, particularly in specimens with a high sperm count?

Answer 64

carbon dioxide lactic acid

Question 65

Despite the limited usefulness of a seminal fluid pH, the measurement is easy to determine and is usually included in a seminal fluid analysis. Commercial pH paper strips with a range from______ should be used and results recorded to the nearest 0.1 pH unit. Appropriate quality control solutions should be used to ensure the accuracy of the pH strips.

Answer 65

4.0 to 10.0

Question 66

Because fructose is produced and secreted by the_____, its presence in semen reflects the secretory function of this gland and the functional integrity of the______ and _____

Answer 66

seminal vesicles ejaculatory ducts and vas deferens.

Question 67

The fructose level is most often determined when the sperm count reveals______ (i.e., no sperm). _________ or _______ ***can cause low fructose levels and azoospermia.***

Answer 67

azoospermia Obstruction of the ejaculatory ducts or abnormalities of the seminal vesicles or vas deferens

Question 68

Normally, semen fructose levels are________. Several quantitative, spectropho-tometric procedures are available for fructose determinations.

Answer 68

equal to or greater than 13 umol per ejaculate

Question 69

A rapid and easy qualitative tube test based on the development of an______ color in the presence of fructose can also be performed With this test, failure of the specimen to develop an _color indicates the absence of fructose. Although this technique is qualitative, relies on the visual assessment of color, and lacks sensitivity to decreased fructose levels, its ease of performance and rapid turnaround time make it a useful tool.

Answer 69

orange-red

Question 70

Quantitative determinations of_____ and ____ in semen can be used to evaluate the secretory function of the prostate gland. The usefulness of zinc and citric acid measurements as markers of biochemical function is ongoing; clinicians are attempting to establish correlations with disease processes (e.g., low zinc levels with prostatitis).

Answer 70

zinc and citric acid levels

Question 71

In normal semen, the total zinc concentration is…

Answer 71

equal to or greater than 2.4 mmol per ejaculate.

Question 72

, the major anion in semen

Answer 72

Citric acid

Question 73

Decreased levels indicate dysfunction of the prostate gland. The total citric acid concentration in normal semen is…

Answer 73

equal to or greater than 52 mmol per ejaculate.

Question 74

is a useful marker to assess the secretory function of the prostate gland. Normally, seminal fluid contains….., whereas other body fluids contain insignificant amounts.

Answer 74

Acid phosphatase activity 200 units of enzyme activity or more per ejacu-late

Question 75

Because of this uniquely high concentration, _______ measurements are often used to determine whether semen is present in vaginal fluid specimens obtained from women after an alleged rape or sexual assault.

Answer 75

prostatic acid phosphatase

Question 76

Even washings of the skin or stained clothing can reveal significant levels of_____, which positively identifies the presence of semen.

Answer 76

prostatic acid phosphatase

Question 77

(2) are being evaluated as indicators of epididymal function, whereas specific ______isoenzymes of sperm are being examined for their clinical use in the evaluation of male fertility.

Answer 77

L-carnitine and a-glucosidase lactate dehydrogenase isoenzymes

Question 77

is a useful marker to assess the secretory function of the prostate gland. Normally, seminal fluid contains….., whereas other body fluids contain insignificant amounts.

Answer 77

Acid phosphatase activity 200 units of enzyme activity or more per ejacu-late